XXIIIrd International Eucarpia symposium, Section Ornamentals ...

XXIII rd International Eucarpia symposium, Section Ornamentals, Leiden, The Netherlands 

XXIII rd International Eucarpia symposium, 

Section Ornamentals 

“Colourful Breeding and Genetics” 

August 31 - September 4 2009 

Leiden 

The Netherlands

XXIII rd International Eucarpia symposium, 

Section Ornamentals 


Welcome 

Dear Colleagues, 

Welcome to the XXIII rd International Eucarpia symposium, section ornamentals “Colourful Breeding and Genetics”, 

organized by Wageningen University and Research Centre in cooperation with Plantum NL and ornamental plant 

breeders of the Netherlands. This symposium aims to be a platform for and to exchange knowledge between scientists 

and plant breeders working on ornamentals from all over the world. This meeting has sessions with oral presentations 

on biodiversity, flower colour, interspecific hybridization, resistance breeding, plant breeder’s rights, breeding and 

genetics, marketing and molecular breeding. In addition to the molecular breeding session there is a Molecular marker 

workshop to inform breeders and scientists on the prospects of molecular assisted breeding. In addition to the oral 

presentations 20 (out of 120) selected posters are presented in 5 minutes in two poster sessions. The historical location 

for the symposium is the beautiful just renewed Stadsgehoorzaal in Leiden, close to the ornamental breeding industry. 

Such a meeting is only possible due to the over 250 participants from more than 35 countries, to the sponsors for their 

financial support and to the organizing committees for the many hours of work. 

We hope that you will enjoy the XXIII rd International Eucarpia symposium, Section Ornamentals “Colourful Breeding and 

Genetics” as well as your stay in Leiden and the Netherlands. 

Jaap van Tuyl 

(Chairman of the Section Ornamentals of EUCARPIA) 

Section Ornamentals

Table of Contents 

1 Organization 1 

Page 

2 Sponsors 3 

3 General information 7 

4 Excursion 9 

5 Programme 11 

6 Abstracts lectures 17 

7 Abstracts posters 74 

8 List of authors 173 

9 List of participants 179 

10 Map of Leiden 199

1. Organization of the XXIII rd International 

Eucarpia symposium, Section Ornamentals 


Convener 

Jaap van Tuyl 

Organizing Committee 

 

 

 

 

 

 

 

 

Jaap van Tuyl, Plant Breeding - Wageningen UR 

Sjaak van Heusden, Plant Breeding - Wageningen UR 

Sjoukje Heimovaara, Royal van Zanten BV 

Kees van ´t Hoenderdal, Dekker Chrysanten BV 

Johan Van Huylenbroeck, ILVO - Belgium 

Wendy ter Laak, Beekenkamp Plants BV 

Ronald Snijder, Royal van Zanten BV 

Thijs Simons, Plantum NL 

Scientific Committee 

 

 

 

 

 

 

 

 

 

Jaap van Tuyl, The Netherlands 

Alain Cadic, France 

Thomas Debener, Germany 

Dik de Vries, The Netherlands 

Sjaak van Heusden, The Netherlands 

Johan Van Huylenbroeck, Belgium 

Antonio Mercuri, Italy 

Masahiro Mii, Japan 

Teresa Orlikowska, Poland 

Organization 1

2. Sponsors 

Main sponsor 

www.syngenta.com 

Sponsor 

www.distel.nl 

www.doriane.com 

Sponsors 3

Sponsor 

www.dekkerchrysanten.nl 

www.fides.nl 

www.iribov.nl 

www.keygene.nl 

www.makbreeding.nl 

www.plantum.nl 

www.rai-worldwide.com 

www.greengenetics.nl 

4 Sponsors

www.deliflor.nl 

www.panamseed.com 

www.dejonglelies.nl 

www.beekenkamp.nl/ornamentals 

www.sandegroup.nl 

www.royalvanzanten.com 

www.bromelia.com 

www.vletterdenhaan.nl 

www.rijnplant.com 

Sponsors 5

3. General information 

Congress Venue 

Stadsgehoorzaal Leiden 

Breestraat 60 

2311 CS Leiden 

The Netherlands 

Congress secretariat 

The registration takes place at the reception desk at the entrance of Breestraat 60. 

The persons dealing with the registration are Niek Botden (only the first day), Mariame Gada (tel 06 234 442 65) 

and Jelle van den Haak (06 242 401 80). 

Catering 

Lunches will be served in the Foyer of the “Aalmarkt” (also called “Kleine Zaal”). 

The symposium dinner will be served in the “Nuon zaal “(also called “Breezaal”). 

Speakers 

Please deliver your presentation as soon as possible at the congress reception desk. 

Lectures 

All lectures will be given in the “Kleine Zaal”. 

Poster sessions 

Poster set-up will start on Monday, August 31, 2009 at 18.00 in the Foyer. 

Please check your poster number in the author’s index. 

Internet access 

For the participants of the congress internet is available in a meeting room on the 3 rd floor. 

General information 7

Proceedings 

The proceedings of this symposium are published in Acta Horticulturae 836 (280 pp, 40 papers; www.actahort.org ). 

During the symposium a number of poster presentations will be selected for publication in a second volume of Acta 

Hortic, the submission deadline will be 15 November 2009). For more information see www.ornamentalbreeding.nl . 

8 General information

4. Excursion 

A choice between the 4 tours should be made in advance, list at the reception desk. 

All Tours from 05.45 till 07.00: 

05.45 Bus leaves from Stadsgehoorzaal (very strictly!) 

07.00 Auction 

Auction Aalsmeer for tours 1 and 3 

Auction Naaldwijk for tours 2 and 4 

09.00 - 15.00 Companies (see below) 

17.00 Hotel 

Tour 1 Aalsmeer region 

Hilverda De Boer (cash&carry) 

Hilverda Kooij (Alstroemeria & Dianthus) 

Florist de Kwakel (Gerbera) 

Tour 2 Westland region 

Fides (Chrysanthemum & pot and bedding plants) 

Beekenkamp Ornamentals (pot and bedding plants) 

NAKTuinbouw (Plant diagnostics & Plant Breeders’ Rights) 

Tour 3 Syngenta, North Holland region 

Syngenta Flower (bedding plants) 

Syngenta Seed (Vegetables) 

Tour 4 Research, Wageningen region 

Plant Science Group (Wageningen University and Research Centre) 

Keygene (Molecular Markers) 

Excursion 9

5. Programme 23 rd EUCARPIA 

SYMPOSIUM - Section Ornamentals 


Monday 31 August 

Welcome reception and registration 

16.00 - 20.00 Registration 

18.00 - 20.00 Welcome reception 

Tuesday 1 September 


08.30 - 08.35 Opening by Jaap van Tuyl 

Introduction and Biodiversity and Convention of Biological Diversity (CBD) treaty 

Chair: Dik de Vries 

08.35 - 09.00 Doeke Faber (Chairman, Dutch Flower Auctions Association, NL) L1 

Ornamental horticulture: where does it end? 

09.00 - 09.30 Orlando de Ponti (President International Seed Federation, ISF) L2 

Access to Biodiversity: New Rules of the Game 

09.30 - 09.50 Birte Lorenzen L3 

Access and benefit-sharing under the CBD - what consequences might an international regime 

have for the horticultural sector? 

09.50 - 10.10 Leila Samiei L4 

In search of genetic variation in Rosa foetida Hermann in Iran 

10.10 - 10.30 Qinglin Liu L5 

Biodiversity and ornamental plant breeding in China 

10.30 - 11.00 Coffee/tea break 

Flower Colour 

Chair: Alain Cadic 

11.00 - 11.30 Yoshikazu Tanaka L6 

The long, winding genetic modification path to more colourful flowers; blue, red and yellow 

11.30 - 11.50 Ellen de Keyser L7 

Flower colour as a model in azalea for integration of phenotype, genotype and gene expression 

11.50 - 12.10 Virginia Gitonga L8 

Inheritance of determinants of flower colour in tetraploid roses 

12.10 - 12.20 Robert Bogers 

International Society for Horticultural Science - Introduction 

12.20 - 13.30 Lunch 

Programme 11

Interspecific hybridization and polyploidy 

Chair: Jaap van Tuyl 

13.30 - 14.00 Masahiro Mii L9 

Breeding of ornamental plants through interspecific hybridization using advanced techniques with 

a special focus on Dianthus, Primula, Cosmos and Kalanchoe 

14.00 - 14.20 Mark Bridgen L10 

Interspecific hybridization of Alstroemeria for the development of new, ornamental plants 

14.20 - 14.40 Tom Eeckhaut L11 

Morphological and anatomical characterisation of chemically induced polyploids in 

Spathiphyllum wallisii 

14.40 - 15.00 Gildas Gâteblé L12 

Advances in Oxera Breeding 

15.00 - 15.20 Valéry Malécot L13 

Polymorphic ITS as a tool to identify hybrids and their parents in cultivated Genisteae (Fabaceae) 

15.20 - 15.40 Ed Morgan L14 

Generating and delivering novelty in ornamental crops through interspecific hybridization: 

some examples 


Short presentations 

Chair: Antonio Mercuri 

16.10 - 16.15 Rodrigo Barba-Gonzalez LP1 

Chromosome identification on the genus Lilium using comparative genomic in situ hybridization 

(CGISH) 

16.15 - 16.20 Matteo Caser LP2 

Discriminating capacity of nucleotide binding site (NBS) and MYB gene profiling for genetic 

analysis of Campanula ecotypes 

16.20 - 16.25 Malgorzata Czernicka LP3 

Verification of the hybrid character of interspecific Rhododendron progeny by molecular tools 

16.25 - 16.30 Emmy Dhooghe LP4 

Production and characterization of intergeneric hybrids between Anemone coronaria and 

Ranunculus asiaticus 

16.30 - 16.35 Marisé Borja LP5 

Breeding system of Glandularia species native to Argentina 

16.35 - 16.40 Yoon-Jung Hwang LP6 

Library construction from micro dissection of chromosome #1 in lily (L. lancifolium) 

16.40 - 16.45 Agnieszka Marasek LP7 

Introgression breeding in genus Tulipa analysed by GISH 

16.45 - 16.50 Jiten Sharma LP8 

Morphological and molecular characterization of intergeneric hybrids between the orchid genera 

Renanthera and Vanda 

16.50 - 16.55 Anta Sparinska LP9 

Diversity in Rosa rugosa x Rosa hybrida interspecific varietie 

16.55 - 17.00 Werachai Tera-arisiri LP10 

Breeding for resistance and biocontrol of wilt disease in Curcuma alismatifolia Gagnep. 

by Bacillus spp. 

17.00 - 17.15 EUCARPIA business meeting 

12 Programme

Wednesday 2 September 


Resistance breeding 

Chair: Teresa Orlikowska 

08.30 - 09.00 Thomas Debener L15 

Current strategies and future prospects of resistance breeding in ornamentals 

09.00 - 09.20 Arwa Shahin L16 

Conversion of molecular markers linked to Fusarium and virus resistance in Asiatic hybrid lilies 

09.20 - 09.40 Carole Koning-Boucoiran L17 

Specific mapping of disease resistance genes in tetraploid cut roses 

09.40 - 10.00 Antra Balode L18 

Breeding for resistance against Botrytis in lily 

10.00 - 10.20 Kullanart Obsuwan L19 

A dysfunctional CymMV movement protein gene confers resistance to CymMV in Dendrobium orchid 


Plant Breeder’s Rights 

Chair: Lidwien Dubois 

10.50 - 11.20 Judith Blokland L20 

Can we still take the breeder’s exemption for granted? 

11.20 - 11.40 Ben Vosman L21 

Essentially derived varieties in ornamentals 

11.40 - 12.00 René Smulders L22 

Analysis of a database of DNA profiles of 734 hybrid tea rose varieties 

12.00 - 12.30 Discussion panel with Judith Blokland, Ben Vosman, Lidwien Dubois 

12.30 - 13.30 Lunch 

Breeding and Genetics 

Chair: Johan van Huylenbroeck 

13.30 - 14.00 Riana Kleynhans L23 

Back to basics for new crop development 

14.00 - 14.20 Neil Anderson L24 

Development of colored, non-vernalization-requiring seed propagated lilies 

14.20 - 14.40 Dik de Vries L25 

Growth and development of cut rose clones; indirect selection for yield 

14.40 - 15.00 Kell Kristansen L26 

In vitro mutagenesis of Aster novibelgii cultivars 

15.00 - 15.20 Lazaro Peres L27 

Breeding the tomato micro-tom model system for ornamental value 

15.20 - 15.40 Leen Leus L28 

Flow cytometry for plant breeding 


Programme 13

Short presentations 

Chair: Sjoukje Heimovaara 

16.10 - 16.15 Pejman Azadi LP11 

A protocol for high rate Agrobacterium-mediated transformation of Lilium 

16.15 - 16.20 Renate Müller LP12 

Transformation with rol-genes of Agrobacterium rhizogenes as a strategy to breed compact 

ornamental plants with improved postharvest quality 

16.20 - 16.25 Antonio Mercuri LP13 

New genotypes of Hibiscus rosasinensis through classical breeding and genetic transformation 

16.25 - 16.30 Youn-Hwa Joung LP14 

Investigation of the factors affecting cross-fertilization rate in rose 

16.30 - 16.35 Supuk Mahattanapuk LP15 

Cloning of the ACC synthase gene from Curcuma alismatifolia Gagnep and its use in 

transformation studies 

16.35 - 16.40 Mohsen Mardi LP16 

Assessing Rosa persica genetic diversity using amplified fragment length polymorphisms analysis 

16.40 - 16.45 Norihiro Ohtsubo LP17 

Redesigning floral architecture: efficient modification of agronomic traits by CRES-T 

16.45 - 16.50 Cristina Borghi LP18 

Kalanchoe x houghtonii: SSH and microarray analysis to screen genes involved in vivipary 

16.50 - 16.55 Luca Pipino LP19 

Pollen characteristics affect seed production of rose cultivars 

16.55 - 17.00 Hanneke Witsenboer LP20 

Application of crops® technology in a wide range of vegetable and field crops 

18.30 - 21.30 Symposium dinner 

Thursday 3 September 

Excursion day 

Friday 4 September 

Marketing 

Chair: Thijs Simons 

08.30 - 09.00 Susanne Lux L29 

Cooperative marketing – a way to stimulate sales and consumption of a plant? 

09.00 - 09.30 Paul Roetenberg L30 

The brand: Frederique’s Choice 

09.30 -10.10 Coffee/tea break 

14 Programme

Molecular Breeding 

Chair: Thomas Debener 

10.10 - 10.30 Olga Shulga L31 

Early-flowering transgenic Chrysanthemum plants 

10.30 - 10.50 Phopgao Buddharak L32 

Isolation and transformation of DFR genes in Curcuma alismatifolia and Clitoria ternatea via 

Agrobacterium tumefaciens 

10.50 - 11.10 Frans Krens L33 

Oriental lily hybrids engineered to resist aphid attack 

11.10 - 11.30 Supatida Sirisawat L34 

DMMADS4, a DEF-like gene from Dendrobium is required for floral organ identity and flower 

longevity of orchid 

11.30 - 11.50 Marina Laura L35 

Over-expression and silencing of KXHKN5 gene in Kalanchoe x houghtonii 

11.50 - 12.10 Marisé Borja L36 

Expression of an Arabidopsis aspartic protease in Pelargonium 

12.10 - 13.30 Lunch 

Workshop: molecular markers and their use in ornamentals 

L37 

13.30 - 13.50 Jan De Riek 

Overview of present use of molecular markers in Ornamental Breeding 

13.50 - 14.10 Paul Arens 

How to develop markers that can be used for identification of ornamental crops 

14.10 - 14.30 Sjaak van Heusden 

What do all the new developments in marker- and sequence technology mean for the use 

of markers in ornamentals? 

14.30 - 14.40 Hanneke Witsenboer 

The potential role of Marker Assisted Selection in breeding varieties 


15.10 - 15.40 Round table discussion with all presenters of workshop 

15.40 - 15.50 Closure by Alain Cadic 

Programme 15

6. Abstracts of lectures and short 

presentations 

ORNAMENTAL HORTICULTURE: WHERE 

DOES IT END? 

Ornamental horticulture has developed over the past 50 years from a local industry to a 

global player. The horticultural sector was in the avant-garde of globalization. Already in 

the late 60’s international trade became an important aspect of the horticultural sector. 

Because flowers and plants are time critical products, trade was initially limited due to 

transportation and temperature control. Present technology allows the movement of 

flowers across the globe by truck, train, boat and airplane. 

The command of the supply chain reversed in the same period. The assortment of 

flowers and plants was largely determined by the growers till the late 90’s, after that 

command in the chain was reversed; consumers determined what was to be produced. 

Over time breeders and growers have learned how to interpret the wishes of the 

consumer; namely, learning from and working with the fashion industry, trend watchers, 

etc, the sector is continually trying to satisfy the ever changing tastes of the consumers. 

Over the past decade, it has been shown that consumer tastes are changing more 

rapidly as regards colours, shapes, types. At the same time, the consumer does not 

want to just buy flowers or plants, he wants to know the story behind the product, and he 

wants to buy sustainably and socially qualified produced flowers. 

This changing behaviour has presented a challenge to breeders and growers alike! 

Innovation is therefore important; not only with regard to shape or colour or smell, but 

also with regard to the internal quality of the flowers or plants as regards disease 

resistance, temperature, growth, etc. 

The marketing and sale of flowers and plants has also been subject to a real change; 

while the flower shop is still the most important outlet in many countries, the market 

share of retail stores, super markets, garden centres and DIY stores have increased 

significantly over the past decade and half! 

Flowers and plants have been an important item in people’s lives. We give flowers at 

birth, illness, marriage, death or other important occasions. 

The challenge for the sector is to ensure that we enlarge the choice for consumers 

by offering a greater assortment of new products and at the same time create more 

occasions to give flowers!. 

L1 

Doeke Faber 

Dutch Flower Auctions 

Association, Aalsmeer, The 

Netherlands 

d.faber@vb.nl 

Session Biodiversity 17

L2 

Orlando de Ponti (1) 

Anke van den Hurk (2) 

(1) Nunhems B.V., Haelen, 

the Netherlands 

O.Deponti@nunhems.com 

(2) Plantum NL, Gouda, the 

Netherlands 

ACCESS TO BIODIVERSITY: NEW RULES OF 

THE GAME 

Plant breeders, including plant breeders of ornamental crops, are dependent on genetic 

resources for the development of improved varieties. In the past the use of genetic 

resources was not regulated, access was free and genetic resources were considered a 

world heritage. Under the Convention on Biological Diversity CBD), ratified in 1993, 

genetic resources were no longer considered a world heritage, but were governed by the 

principle of national sovereignty. This resulted in entirely new basic rules for Access and 

Benefit Sharing (ABS), which still need to be further elaborated and implemented. 

In 2004 the International Treaty on Plant Genetic Resources for Food and Agriculture 

(IT) was ratified. A so-called multilateral system for a limited list of plant species for food 

and agriculture is part of the IT. The ABS arrangements for the genetic resources that 

are part of the multilateral system are regulated through a Standard Material Transfer 

Agreement, which was approved in 2006. Through this agreement access is possible 

under reasonable standard benefit sharing arrangements. Most importantly, the 

breeders’ exemption is recognized as a benefit on its own, and in that case mandatory 

payments to the IT for the use of the genetic resources are not required. 

The species not falling under the multilateral system of the IT, including all 

ornamental species, fall automatically under the regime of the CBD. So, legal access to 

genetic resources of ornamental species can only be managed by bilateral agreements 

with the relevant national competent authorities of each specific country. 

Legal access to valuable genetic resources for plant breeding not falling under the 

multilateral system of the IT is complex, if not impossible. However, both private and 

public breeders are responsible to take the necessary measures to have legal access to 

genetic resources, and possibly have to accept that some genetic resources are not (yet) 

available. As an international regime does not yet exist, ornamental breeders should 

strongly advocate for a workable ABS regime, which will re-establish efficient and fair 

access to these valuable resources. 

18 Session Biodiversity

ACCESS AND BENEFIT-SHARING UNDER THE 

CBD - WHAT CONSEQUENCES MIGHT AN 

INTERNATIONAL REGIME HAVE? 

The international negotiations on an International Regime on Access and Benefit- 

Sharing are making progress; it is planned to adopt a system in 2010. Even though 

meanwhile at least part of the negotiators see a need for tailored solutions for different 

industries, horticulture and agriculture are held to have the same interests. But is this 

really the case? CIOPORA developed a position on the specialities and needs of 

breeders of asexually reproduced ornamental and fruit varieties. 

We describe how access and benefit-sharing is carried out in the horticultural sector, 

what is different about the sector compared to others and show how a solution should 

look like for our sector. The basic assumption is, that by legal and practical means 

already without any additional administrative instruments sufficient benefit-sharing is 

executed and that a model like under the ITPGRFA might not be first choice for breeders 

of vergetatively reproduced ornamental and fruit varieties. 

L3 

Birte Lorenzen 

CIOPORA, Gänsemarkt 45, 

20354 Hamburg, Germany 

Birte.Lorenzen@ciopora.org 


L4 

Leila Samiei (1) 

Roohangiz Naderi (1) 

Ahmad Khalighi (1) 

Ali-Akbar Bushehri (2) 

Valiollah Mozaffarian (3) 

Marinus JM Smulders 

(4) 

Danny Esselink (4) 

(1) Dept. of Horticulture, 

College of Agriculture and 

Natural Resources, Tehran 

Univ., Iran 

(2) Dept. of Biotechnology, 

College of Agriculture and 

Natural Resources, Tehran 

Univ., Iran 

(3) Research Institute of 

Forest and Rangelands, 

Tehran, Iran 

(4) Plant Research 

International, Wageningen 

UR, P.O. Box 16, 6700 AA 

Wageningen, The 

Netherlands 

leilisamie@yahoo.com 

IN SEARCH OF THE GENETIC VARIATION IN 

ROSA FOETIDA IN IRAN 

Iran is considered as one of the major centers of plant biodiversity. There are very 

diverse natural environments that lead to the high genetic diversity in plants. Rosa 

foetida, also known as Persian yellow briar, is one of the important species amongst 13 

rose species that occur in Iran. It is a dense erect shrub (up to 2 m) with bright yellow or 

scarlet flowers with a yellowish reverse petal. It is most abundant in South West Asia. In 

Iran R. foetida occurs mainly in the north and west regions, notably in Tehran, West 

Azarbaijan and Kurdestan provinces, which are mountainous and contain very diverse 

habitats. In addition, some plants can be found in Eastern and Southern Iran. It is 

reported that this species is the strong yellow colour in modern roses. In this study we 

have used 10 microsatellite markers to determine diversity in Rosa foetida accessions 

collected across Iran. The dendrogram obtained using dice similarity coefficient resulted 

in only 2 genotypes. To our surprise, most of the samples collected showed the same 

genotype, even if they were collected at different sites, and only two accessions were 

representative the other genotype. The results are discussed in relation to breeding 

system, human influence and overall gene pool status. 

20 Session Biodiversity

BIODIVERSITY AND ORNAMENTAL PLANT 

BREEDING IN CHINA 

China has a flower growing history of more than 2000 years, but the floricultural industry 

just began from the 1985 onwards. Floricultural industry not only contributes to enrich 

the people’s spiritual life and to improve the living environment, but also is an important 

way to regulate the planting structure and to increase the farmers’ revenue in China. 

Cultivars are the basic material of flower production. If only the new cultivars without 

Chinese breeder’s rights were produced, it is no real sense to the Chinese floricultural 

industry. Although there are more than 5600 species of ornamental plants distributed in 

China, and 4400 cultivars bred in China, the commercial cultivated floricultural crops are 

mostly foreign species and cultivars. There is plenty of ornamental germplasm in China, 

but the often-used breeding materials are cultivars instead of species. Many ornamental 

species were bred in China, which mainly concentrated on some traditional famous 

flowers. However, pot plants, bedding flowers and woody garden plants are often 

involved. About 100 new flower cultivars were released annually, but few of them 

were used large-scale in floriculture or landscaping. Presently, the main problems to 

flower breeding in China are who should be the breeders and how to protect breeder's 

rights. 

L5 

Hongqiang Yu 

Ying Feng 

Qinglin Liu 

Department of Ornamental 

Horticulture and Landscape 

Architecture, Agricultural 

University, Beijing 100193, 

China 

liuql@cau.edu.cn 


L6 

Yoshikazu Tanaka (1) 

Steve Chandler (2) 

(1) Suntory Ltd. 1-1-1 

Wakayamadai, Shimamoto, 

Mishima, Osaka 618-8503, 

Japan 

(2) Florigene Pty Ltd. 1 

Park Drive, Bundoora, 

Victoria 3083, Australia 

Yoshikazu_Tanaka@ 

suntory.co.jp 

THE LONG, WINDING GENETIC 

MODIFICATION PATH TO MORE COLOURFUL 

FLOWERS: BLUE, RED AND YELLOW 

Flower breeding is constrained by the limited gene sources available within a target 

species. It is rare for a single species to have all possible flower colours, due to the lack 

all genes on the pigment biosynthesis pathway. Molecular breeding utilizing genetic 

engineering techniques has liberated breeders from this gene-pool constraint. For 

successful molecular breeding, it is necessary to isolate relevant genes, establish 

transformation systems, optimize expression of transgenes and obtain regulatory 

permission for both production and consumption. Though all these procedures are very 

often expensive and time consuming, the investment is long-term and a financial return 

is possible for transgenic flowers that meet the consumer’s demand.. 

We have been developing transgenic flowers, with modified flower colour, for many 

years. Flavonoids and the coloured sub-class of compounds, anthocyanins, are 

dominant colour constituents of most flowers. The flavonoid biosynthetic pathway has 

been well studied and most biosynthetic genes have been obtained. It is feasible to 

generate white, yellow, red and blue flowers by engineering the pathway; both by overexpression 

of heterologous genes and/or down-regulation of endogenous genes. 

The major cut-flower species rose, carnation and chrysanthemum, lack blue/violet 

cultivars because they do not accumulate the delphinidin-based anthocyanins that are 

present in most blue/violet flowers. Flavonoid 3’,5’-hydroxylase (F3’5’H) is the critical 

enzyme for delphinidin biosynthesis and this gene is missing in species that do not 

accumulate delphinidin-based anthocyanins. An efficient transformation system was 

developed for rose and carnation and after introduction of the gene encoding F3’5’H 

transegnics were produced whose flowers accumulated delphinidin-based anthocyanins 

and an altered colour. Through careful choice of host cultivar and optimization of the 

expression of transgenes it has been possible to obtain flowers which accumulate 

delphinidin-based anthocyanins only. Such flowers exhibit an attractive colour change to 

blue/violet. Transgenic colour-modified carnation has been available commercially since 

1996 and the first commercially available, colour-modified rose variety will be sold in 

2009. From a biosafety perspective the transgenic flowers lines have no impact on the 

environment or health and pose negligible, if any, risk to biodiversity. 

22 Session Flower colour

FLOWER COLOUR AS A MODEL IN AZALEA 

FOR INTEGRATION OF PHENOTYPE, 

GENOTYPE AND GENE EXPRESSION 

Flower colour is inherited as a semi-qualitative trait in azalea and is mainly determined 

by differences in anthocyanins and flavonols. A two-gene model is used to explain the 

phenotypic variation between white, brick red and carmine red colour: W in case the 

flower petals contain anthocyanins and Q if flavonols are present as co-pigments. 

However, the presence of flavonols in white flowers cannot be detected visually. Also, 

the existence of pink flowers is not explained by this two-gene model. Therefore, flower 

colour was determined on a crossing population using image analysis software and 

discriminant analysis was used for classification. Integration of the image analysis data 

as QTLs on a genetic map of the crossing population could be enlightening. A genetic 

map of 16 linkage groups was constructed. Besides anonymous AFLP and SSR markers 

also a set of functional markers were used. EST-markers were developed for four genes 

coding for key-enzymes in the flavonoid biosynthesis pathway. MYB-profiling, a 

sequence directed technique similar to NBS-profiling, generated in this crossing 

population fifteen dominant markers functionally related to the MYB gene family. In this 

way, phenotypic and genetic data were both integrated on the genetic map of azalea. In 

case both are located at the same mapping position, these genes are proven to be 

directly involved in the creation of the phenotypic variation of the trait. Nevertheless, it is 

very likely that not the genes themselves but transcription factors are the switches that 

regulate the phenotype of the trait. In that case, phenotype and genotype will be mapped 

at different positions, but phenotype is then expected to be mapped together with the 

true regulators, the transcription factors e.g. MYB markers. To confirm this theory, gene 

expression profiles of five flavonoid biosynthesis genes were generated in petals of a 

selection of flowers of the crossing population using qPCR and eQTL mapping again 

integrated these data with the genetic map. 

L7 

Ellen De Keyser (1) 

Peter Lootens (1) 

Jan De Riek (1) 

Erik Van Bockstaele 

(1,2) 

(1) Institute for Agricultural 

and Fisheries Research 

(ILVO), Plant Sciences 

Unit, Applied Genetics and 

Breeding; Caritiasstraat 21, 

9090 Melle, Belgium 

(2) Gent University, Faculty 

of BioScience Engineering, 

Dept. of Plant Production; 

Coupure links 653, 9000 

Gent, Belgium 

ellen.keyser@ilvo. 

vlaanderen.be 

Session Flower colour 23

L8 

Virginia Gitonga 

Robert Stolker 

Simon Ribot 

Carole Koning- 

Boucoiran 

Frans Krens 

Wageningen University and 

Research Center, Plant 

Breeding, P.O. Box 16, 

6700 AA Wageningen, The 

Netherlands 

virginia.gitonga@wur.nl 

INHERITANCE OF DETERMINANTS OF 

FLOWER COLOUR IN TETRAPLOID ROSES 

The choice of selection breeding for crop improvement in rose requires a better 

understanding of biological mechanisms and knowledge of the inheritance of the major 

target traits which can lead to new or improved screening methods. One of the problems 

in cut roses is that flower colour of some genotypes is not stable across growing 

environments. The dependency upon genotype suggests that this is a heritable trait. In 

general in rose, the genetic knowledge is still limited. Wageningen UR Plant Breeding 

together with international partners has produced one of the rose diploid maps and work 

is currently going on to create a highly dense tetraploid map using the progeny (181 

individuals) from a cross between two tetraploid rose genotypes (P540 and P867) made 

available by Terra Nigra b.v.. The map will contain both phenotypical traits as well as 

molecular markers. The two parents were chosen to ensure sufficient genetic variation 

and segregation in the progeny for many morphological traits including colour, but also 

for disease resistance/susceptibility. 

The current mapping population will be characterized for flower colour, by using 

colour charts such as the official chart of the Royal Horticultural Society, and additionally, 

by image analysis and measuring reflectance using a spectrocolorimeter. The genetics 

of flower colour will be determined by QTL analysis. In addition, flower petals of all 

genotypes will be analysed by HPLC to characterize secondary metabolic components 

that determine flower colour, such as anthocyanins. The inheritance of these 

components will also be assessed and compared to that of flower colour. 

24 Session Flower colour

BREEDING OF ORNAMENTAL PLANTS 

THROUGH INTERSPECIFIC HYBRIDIZATION 

USING ADVANCED TECHNIQUES WITH A 

SPECIAL FOCUS ON DIANTHUS, PRIMULA, 

COSMOS AND KALANCHOE 

In ornamental plants, interspecific hybridization has successfully been used to produce 

novel cultivars with useful traits of both parents and to incorporate desirable traits of one 

species to another. Advanced breeding techniques like embryo rescue, polyploidization, 

protoplast fusion and molecular cytogenetic methods are used to produce and 

characterize interspecific hybrids in various taxonomic groups.. In this presentation, 

recent advances and problems in interspecific hybridization are described with special 

references to the use of embryo culture techniques for rescuing the abortive hybrid 

embryos and to the use of artificial polyploidization techniques for restoring the fertility of 

the interspecific hybrids. Some interesting topics related to interspecific hybridizations 

are also described such as the production of unexpected ploidy plants due to unreduced 

gamete formation and spontaneous chromosome doubling during in vitro culture in 

Dianthus, Primula, Cosmos and Kalanchoe. Some of the products of these interspecific 

hybridizations have successfully been commercialized directly or used as the 

germplasms for further breeding of novel ornamental crops. 

L9 

Masahiro Mii 

Graduate School of 

Horticulture, Chiba 

University, 648 Matsudo, 

Matsudo, Chiba 271-8510, 

Japan 

miil@faculty.chiba-u.jp 

Session Interspecific hybridization 25

L10 

Mark Bridgen (1) 

Elizabeth Kollman (2) 

Chunsheng Lu (3) 

(1) Cornell University, 3059 

Sound Ave., Riverhead, NY 

11901 USA 

(2) Four Star Greenhouse, 

1015 Indian Trails, 

Carleton, MI 48117 USA 

(3) Plant Sciences, Inc., 

342 Green Valley Rd., 

Watsonville, CA 95076 

USA 

mpb27@cornell.edu 

INTERSPECIFIC HYBRIDIZATION OF 

ALSTROEMERIA FOR THE DEVELOPMENT OF 

NEW, ORNAMENTAL PLANTS 

Alstroemeria, the Inca Lily or Lily-of-the-Incas, is a popular cut flower plant because of 

the wide variety of flower colours that are available and the long postharvest life of its cut 

flowers. In recent years, cold-hardy introductions (USDA winter hardiness zone 5) by the 

authors have expanded the interest of this everblooming plant as a garden perennial. 

This research describes breeding procedures that have been used with the objective to 

breed novel, commercially valuable cold-hardy and fragrant flowered cultivars of 

Alstroemeria. Winter-hardy hybrids were developed by using the Chilean species, 

Alstroemeria aurea and fragrant hybrids were developed by using the Brazilian species, 

Alstroemeria caryophyllae. Interspecific hybrids were bred with the assistance of in vitro 

techniques such as in ovulo embryo rescue, micropropagation, and somatic 

embryogenesis. For embryo rescue, ovaries were collected 10-23 days after hand 

pollination and their ovules were aseptically excised. Ovules were placed in vitro on 25% 

Murashige and Skoog (MS) medium under dark conditions until germination. Three 

weeks after germination they were subcultured onto full-strength MS medium with 2 mg 

benzyl adenine (BA) per liter. Subsequently, plants were subcultured every three to four 

weeks onto a liquid MS medium with BA until they were large enough for rooting. After 

rooting and acclimation, plants were transferred to the greenhouse. Successful hybrids 

were evaluated under both greenhouse and field trials to determine winter survival and 

performance. As new hybrids and cultivars were developed, research was completed to 

develop production and propagation protocols for the plants. As a result of this research, 

the only fragrant cultivar of Alstroemeria, ‘Sweet Laura’, was developed. The new 

Alstroemeria cultivars ‘Mauve Majesty’ and ‘Tangerine Tango’ were recently patented 

and introduced by Cornell University. These plants are noteworthy by their winter 

hardiness, everblooming growth habit, and long flower stems. This project demonstrates 

the enormous potential for new plant development that exists. 

26 Session Interspecific hybridization

MORPHOLOGICAL AND ANATOMICAL 

CHARACTERISATION OF CHEMICALLY 

INDUCED POLYPLOIDS IN SPATHIPHYLLUM 

WALLISII 

Tetraploids were induced in Spathiphyllum wallisii Regel (2n = 2x = 30) through in vitro 

application of mitosis inhibitors. Tetraploids were compared to the original diploid control 

plants. Polyploidization had a significant effect on plant anatomy and morphology. The 

stomatal area of diploids was smaller compared to the tetraploid plants. The leaf angle 

was smaller in diploids. The stomatal length and width, leaf thickness and angle and 

thickness of the spathum were positively correlated to the higher ploidy level. On the 

other hand, stomatal density, length/width ratio of leaf, spathum and spadix, number of 

shoots and leafs and length of the flower stalk decreased in tetraploids compared to the 

corresponding diploid controls. The leaf number of diploid plants was higher compared to 

tetraploids. Altogether, this study quantified the extended morphological changes of 

chromosome doubling in our model crop Spathiphyllum wallisii. 

L11 

Ives Vanstechelman 

(1,2) 

Hein Vansteenkiste 

(3)einH,jjjjj 

Tom Eeckhaut (1) 

Johan Van 

Huylenbroeck (1) 

Marie-Christine Van 

Labeke (2,4) 





Breeding, Caritasstraat 21, 


(2) Ghent Univ., 

Department of Plant 

Production, Coupure links 

653, 9000 Gent, Belgium 

(3) Research Center for 

Ornamental Plants (PCS), 

Schaessestraat 18, 9070 

Destelbergen, Belgium 

(4) Faculty Biosciences and 

Landscape Architecture, 

University College Ghent - 

Ghent University 

Association, Voskenslaan 


tom.eeckhaut@ilvo. 

vlaanderen.be 


L12 

Gildas Gâteblé 

Virginie Lemay 

Jacqueline Ounémoa 

Institut Agronomique néo- 

Calédonien, Station de 

Recherche Agronomique 

de Saint Louis 

BP 711. 98810 Mont Dore. 

New Caledonia 

gateble@iac.nc 

ADVANCES IN OXERA BREEDING 

The New Caledonian sub-endemic Oxera genus comprises several species of great 

ornamental interest. In 2006, we started to set up a vast interspecific hybridization 

program in order to create new cultivars with combined ornamental features. Twenty one 

of the twenty two endemic taxa (O. neriifolia subsp. sororia was not found during several 

field searches) were used in this breeding program and more than 7600 hand pollination 

crosses were performed. Pollinations were done with freshly collected pollen prior to bud 

opening and after immature anthers removal. 303 fruits containing 566 seeds were 

harvested and sown giving 361 viable plants. Limited by available plants and flower 

buds, 80 combinations out of the 420 theorically possible ones were trialled. Seeds were 

obtained in 37 combinations while 27 combinations gave viable plants. Only three taxa 

(O. crassifolia, O. glandulosa and O. morierei) involved in these crosses did not give any 

offspring. Oxera sulfurea, a species which flowers throughout the year, as the female 

parent, was involved in 3294 cross pollinations and has generated 284 offspring from 15 

of the 20 taxa trialled. All the putative hybrids produced have intermediate morphological 

vegetative characters between the two parents and the ones that have already flowered 

also present intermediate flower forms and colours. This breeding program is still 

ongoing and cytology work is underway to determine the basic number of chromosomes 

of the genus Oxera and to learn if there are any different ploidy levels between the 

species. 


POLYMORPHIC ITS AS A TOOL TO IDENTIFY 

HYBRIDS AND THEIR PARENTS IN 

CULTIVATED GENISTEAE (FABACEAE) 

Internal Transcribed Spacer (ITS) of ribosomal DNA is a classical sequence used for 

phylogenetic analysis. Usely, concerted evolution homogenise the numerous ITS of an 

organism in such way that a single sequence is amplified. However, for vegetatively 

propagated plants of hybrid origin, several ITS sequences can be obtained from a 

single individual. By cloning ITS sequences of various taxa assigned to tribe Genisteae 

within Fabaceae, we have been able to show such ITS polymorphism in Cytisus x 

kewensis, C. x dallimorei, C. x praecox, C. “racemosus” and C. ‘Porlock’ among others. 

For these taxa, the distinct ITS sequences can be compared with sequences of 

putative parents. We have then confirmed that Cytisus x kewensis is a C. ardoinoi X C. 

multiflorus cross. C. “racemosus”, commonly presented as a C. canariensis X C. 

stenopetalus and thus named C. x spachianus may in fact be a more complex hybrid 

involving also C. monspessulanus. We have also confirmed that C. ‘Porlock’ is a C. 

monspessulanus X C. “racemosus”. We thus propose a way to asses the interspecific 

hybrid status of any vegetatively propagated plant, that also allows to clearly identify 

the putative parent taxa. This confirm a feature observed also in other wild and 

cultivated groups such as Viola, Paeonies and Amelanchier. Using additional data, 

mainly chloroplast microsatellites, we can further identify the maternal parent. These 

additional tools has shown that Cytisus x kewensis ‘Niki’ is a C. ardoinoi X C. 

multiflorus cross with C. multiflorus as maternal parent, while Cytisus x kewensis is a 

result of the same cross but with C. ardoinoi as maternal parent. 

L13 

Valéry Malécot 

Nadège Macquaire 

Gaëlle Auvray 

Véronique Kapusta 

UMR 1259 GenHort , INRA, 

AGROCAMPUS-OUEST, 

Univ. Angers 

2 rue le Nôtre, 49045 Angers 

Cedex 01, France 

valery.malecot@agrocampusouest.fr 


L14 

E R Morgan (1) 

G K Burge (1) 

G Timmerman-Vaughan 

(2) 

M Debenham (1) 

(1) New Zealand Institute 

for Plant & Food Research 

Limited, Private Bag 11 

600, Palmerston 

North 4442, New Zealand 

(2) New Zealand Institute 

for Plant & Food Research 

Limited, Private Bag 

4704, Christchurch 8140, 

New Zealand 

morgane@crop.cri.nz 

GENERATING AND DELIVERING NOVELTY IN 

ORNAMENTAL CROPS: SOME EXAMPLES 

The generation of novelty is a key focus for breeders of ornamental crops. This paper 

describes the application of a range of in vitro techniques to generate novel genetic 

combinations from which new varieties can be selected. 

A program of interspecific hybridisation between Limonium sinuatum and L. perezii 

has resulted in a significant number of hybrids and back cross hybrids. All hybrids 

produced to date have had either very low or complete infertility and require 

chromosome doubling to restore fertility. A feature of the new hybrids is the increased 

visual impact of flowering stems conferred by branching angle and increased space 

between individual florets in the inflorescence. 

Sandersonia is a flower crop well known to New Zealand growers. The single 

species has shown very little variation despite being entirely seed propagated. Attempts 

to increase variation in this crop have included hybridisation with related genera such as 

Littonia and Gloriosa. Gloriosa superba is a very variable species featuring individuals 

with ploidy levels from diploid to octoploid. There are considerable differences in the 

performance of hybrids between Sandersonia and Gloriosa with some lines almost 

impossible to grow and others extremely vigorous. Hybrids have all been infertile and to 

date chromosome doubling has not been successful in any of the lines tested. 

Gentiana is a relatively poorly known cut flower crop. Ten years ago the colour range 

consisted of blues, pinks and whites. Red-flowered varieties are now available to 

commerce. A current challenge is to generate yellow and orange forms in order to 

provide a complete colour range. Yellow-flowered interspecific hybrids have been 

developed between G. triflora and G. lutea. These hybrids have proven difficult to grow 

out of in vitro culture. Chromosome doubling has been carried out to overcome the 

anticipated infertility of these hybrids, and work is underway to develop a strategy for 

producing back cross hybrids 


CHROMOSOME IDENTIFICATION ON THE 

GENUS LILIUM USING COMPARATIVE 

GENOMIC IN SITU HYBRIDIZATION (CGISH) 

Single chromosome identification is of primal importance in the study of evolutive 

process in complex genomes, such as polyploidization and hybridization events, among 

others. Traditionally, chromosome identification is made by arranging the chromosomes 

by the length of the short (p) and the long (q) arms, identifying the centromeres and 

secondary constrictions. Furthermore, accurate chromosome identification can be 

performed trough chromosome differentiation techniques such as C-, N-, and Q-banding. 

In the last years, through the development of Fluorescent In Situ Hybridization 

techniques (FISH), even more accurate single chromosome identification has been 

accomplished, by the hybridization of highly conserved repetitive sequences, such as 

rDNAs. However, particular probes must be developed through laborious isolation and 

cloning molecular techniques. The comparative genomic in situ hybridization (cGISH) is 

a straightforward technique that allows the identification of single chromosomes by the 

generation of signals of conserved DNA regions along the chromosomes of different 

species. In this study we labeled total genomic DNA of Triticum aestivum and 

Arabidospis thaliana and hybridized it to chromosomes of different species of the genus 

Lilium. Different stringencies were applied to determine the optimum removal of cross 

hybridization, the 80% stringency showed to be the best, giving a clear signal and 

removing most of the cross hybridization. Triticum aestivum total genomic DNA exhibited 

six landmarks on three homologous chromosomes in the three different cultivars while 

Arabidopsis thaliana total genomic DNA exhibited six landmarks on three homologous 

chromosomes on Lilium, one of these signals being in a different chromosome of those 

of the T. aestivum signals. Together with the DAPI bands the total genomic DNAs 

landmarks allowed the identification of six out of 12 single homologous chromosomes. 

LP1 

Rodrigo Barba-Gonzalez 

Centro de Investigación y 

Asistencia en Tecnología y 

Diseño del Estado de 

Jalisco A.C. 

Av. Normalistas #800 

Colinas de la Normal. 

Guadalajara, Jalisco. 

México CP 44270 

rbarba@ciatej.net.mx 

Session Short presentations 31

LP2 

M. Caser (1) 

V. Scariot (1) 

P. Arens (2) 

(1) Department of 

Agronomy, Forest and Land 

management, Università 

degli Studi di Torino, Via 

Leonardo da Vinci 44, I- 

10095 Grugliasco, Torino, 

Italy 


International, Wageningen 

UR, Droevendaalsesteg 1, 

PO Box 16, NL-6700 AA 

Wageningen, the 

Netherlands 

matteo.caser@unito.it 

DISCRIMINATING CAPACITY OF NUCLEOTIDE 

BINDING SITE (NBS) AND MYB GENE 

PROFILING FOR GENETIC ANALYSIS OF 

CAMPANULA ECOTYPES 

Two DNA-based marker methodologies, nucleotide binding site (NBS) and myb gene 

profiling, were evaluated with respect to their discriminating capacity and efficiency in 

genetically analyzing 11 Campanula ecotypes belonging to C. latifolia (Lat1 and Lat2), C. 

rapunculoides (Rap1, Rap2, Rap3 and Rap4), C. spicata (Spic1, Spic2 and Spic3) and 

C. trachelium (Trach1 and Trach2). Three restriction enzymes (MseI, RsaI and HaeIII) 

were utilized with two NBS primers (GLPL6 and NBS2) and one myb primer (MYB2). 

Looking at the number of banding pattern, all two techniques discriminated the 

genotypes very effectively. On an individual assay basis, NBS profiling completely 

distinguished all the genotypes of the ecotypes Lat1 (C. latifolia) and Trach1 (C. 

trachelium). To assess the usefulness of the overall information provided by these 

marker data for establishing phylogenetic relationships, cluster analysis was performed. 

For the two markers a high similarity in dendrogram topologies was obtained although 

some differences were observed. In general, the dendrograms reflected the 

geographical diffusion of the ecotypes. The myb marker resulted to be more adept to 

differentiating species rather than ecotypes, while NBS profiling appeared to be more 

able to discriminate the ecotypes. Comparable population structuring was obtained with 

both marker systems. Two Bayesan assignment tests with the program STRUCTURE 

divided the accessions into groups agreeing with neighbour-joining trees. Both NBS and 

myb gene profiling data were demonstrated to be remarkably effective for group 

discrimination and genetic diversity studies. The use of these systems is discussed in 

terms of the choice of appropriate marker techniques for different aspects of local 

Campanula ecotypes evaluation. 

32 Session Short presentations

VERIFICATION OF THE HYBRID CHARACTER 

OF INTERSPECIFIC RHODODENDRON 

PROGENY BY MOLECULAR TOOLS 

Progeny derived from five types of interspecific rhododendron crosses, i.e. (1) R. aureum 

x R. brachycarpum, (2) R. aureum x ‘Catharine van Tol’, (3) R. aureum x R. 

yakushimanum ‘Koichiro Wada’, (4) R. yakushimanum ‘Koichiro Wada’ x R. aureum and 

(5) ‘Nova Zembla’ x R. aureum, were investigated to confirm their hybrid character. The 

verification was based on genomic in situ hybridization (GISH) and supported by the 

RAPD-PCR method. Owing to the fact that GISH has never been used in analysis of 

rhododendrons, thorough optimization of the protocol was necessary. We assayed one 

to five plants representing each cross. Microscopic preparations of mitotic chromosomes 

of the tapetum and meiotic chromosomes of pollen mother cells (PMCs) were prepared 

by enzymatic digestion of anthers. Total genomic DNA from pollen donors was used as a 

probe in the ratio of 3 ng/μl and total genomic DNA from maternal plants was used as a 

blocking DNA (60-fold excess of the labeled genomic probe). The developed GISH 

protocol in rhododendron, has enabled the identification of parental chromosomes in 

genomes of the studied progeny. Hybridization signals, indicating the localization of the 

probes on chromosomes and on interphase nuclei, were detected in four interspecific 

rhododendron F1 hybrids. Obtained results were confirmed by RAPD-PCR analysis. 

For this purpose DNA was amplified with 19 RAPD primers and we obtained RAPD 

markers which allowed to describe the relationship between parents and their progeny. 

None of studied plants representing the cross ‘Nova Zembla’ x R. aureum, was proven to 

be a hybrid. Our study confirmed that both genomic in situ hybridization (GISH) and 

RAPD-PCR markers were powerful tools for verification of the hybrid character of the 

rhododendron progeny derived from wide crosses. 

LP3 

Małgorzata Czernicka 

Anna Nowicka 

Ewa Grzebelus 

Dariusz Grzebelus 

Maria Klein 

Departament of Genetics, 

Plant Breeding and Seed 

Science,University of 

Agriculture in Krakow, Al. 

29 Listopada 54, 31-425 

Kraków, Poland, 

czernickam@bratek. 

ogr.ar.krakow.pl 


LP4 

E. Dhooghe 

D. Reheul 

M-C. Van Labeke 

Department of Plant 

Production, Faculty of 

Bioscience Engineering, 

Ghent University, Coupure 

links 653, B-9000 Gent, 

Belgium 

emmy.dhooghe@ugent.be 

PRODUCTION AND CHARACTERIZATION OF 

INTERGENERIC HYBRIDS BETWEEN 

ANEMONE CORONARIA AND RANUNCULUS 

ASIATICUS 

Ranunculus asiaticus L. and Anemone coronaria L. are common cut flowers which 

belong to the family of the Ranunculaceae. Between these species a high degree of 

variation can be found in leaves, flower shape and flower colour. Therefore intergeneric 

crossings between these genera might result in new interesting hybrids. Many of such 

wide hybridizations do not occur naturally because prezygotic and postzygotic barriers 

alter fertilization, embryogenesis and/or seed formation. However elaborate in vitro and 

in vivo work can overcome these barriers. The objective of this study was to produce and 

characterize hybrids between Anemone coronaria and Ranunculus asiaticus. 

Crosses between Anemone and Ranunculus were performed in the greenhouse from 

January till May. Three to four weeks after pollination immature seeds were harvested 

and rescued in vitro. One year or two years later, parents and F1 progeny were grown in 

identical growth conditions in order to study possible morphological differences. To 

distinguish the plants on molecular level AFLP analysis was performed on F1 progenies 

and parents using the restriction endonuclease EcoRI as suggested by Nissim et al. 

2004. 

In most cases, crosses between Ranunculus and Anemone (and vice versa) were 

prevented because the pollen tube was not able to reach the ovules. The breeding 

experiments resulted therefore in a very poor seed set. The phenotype of the hybrids 

obtained following embryo rescue, showed a high similarity of that of the mother plant. 

However some novel characteristics were observed. Visual screening revealed variation 

in flower colour. Furthermore AFLP analysis demonstrated that some bands of the 

mother plant were deleted in the hybrids. Also some specific bands of the father plant 

and novel bands, absent in mother as father, were observed. 

In conclusion, we were able to produce intergeneric hybrids between Anemone and 

Ranunculus. These hybrids have a similar morphology compared to the mother plant but 

the AFLP results showed some genomic reorganization. This is in accordance with other 

intertribal crossing studies, showing similar phenotypical and molecular phenomenons. 

To confirm these (preliminary) results, independent techniques such as chromosome 

spreads and GISH, analyzing chromosomal reorganisation, will be used in the future. 

References: 

Nissim Y, Jinggui F, Arik S, Neta P, Uri L, Avner C (2004) Phenotypic and genotypic 

analysis of a commercial cultivar and wild populations of Anemone coronaria. Euphytica 

136:51-62. 


BREEDING SYSTEM OF GLANDULARIA 

SPECIES NATIVE TO ARGENTINA 

The Glandularia peruviana, G. platensis and G. glandulifera breeding system was 

studied using controlled manual pollination. Self-pollination and cross-pollination 

experiments were conducted in selected clones under greenhouse conditions. Natural 

pollination was also studied as control. Pollen tube growth was analysed in self and 

cross- pollinated pistils to determine the existence of an incompatibility system. 

Self-pollinated flowers produced no fruits in all species. Cross-pollinated flowers 

produced 46%, 8% and 80% fruits in G. peruviana, G. platensis and G. glandulifera 

respectively. 

In the three species, pollen grains germinated normally in stigma in self and cross 

pollinated pistils, but further growth of the pollen tube was inhibited at different parts of 

the style in self pollinated pistils. Pollen tube growth rate is discused for both pollination 

conditions. 

The behaviour of pollen tube growth and fruit set production experiments indicate the 

presence of a gametophytic self-incompatibility system which is characterized by the 

inhibition of the pollen tube growth at the style. 

LP5 

L. Imhof (1) 

M. Borja (2) 

G. Facciuto (3) 

(1) Universidad Católica de 

Córdoba, Argentina 

(2) Universidad 

Complutense de Madrid, 

F. Promiva, España 

(3) Instituto de Floricultura, 

INTA, Argentina 

leimhof@hotmail.com 


LP6 

Yoon Jung Hwang (1) 

In-Suk Ahn (1) 

In Sook Park (1) 

Si-Yong Kang (2) 

Jae-Dong Chung (1) 

Ki Byung Lim (1) 

(1) School of Applied 

Science, Kyungpook 

National University, Daegu 

702-701, Korea 

(2) Dept. of Radiation Plant 

Breeding and Genetics, 

Advanced Radiation 

Technology Institute, Korea 

Atomic Energy Research 

Institute, Geongeup 580- 

185, Korea 

wowyuki@hanmail.net 

LIBRARY CONSTRUCTION FROM 

MICRODISSECTION OF CHROMOSOME #1 IN 

LILY (L. LANCIFOLIUM) 

Lilium lancifolium belongs to section Sinomartagon and has been known as polyploidy 

complex comprising diploid (2n=2x=24) and triploid (2n=3x=36). Chromosome #1 is the 

longest chromosome as a metacentric in the L. lancifolium metaphase complements. It 

was microdissected and collected from mitotic metaphase spreads of lily (L. lancifolium) 

by using PALM Robot MicroBeam System (Carl Zeiss AG, Germany), attached to an 

Axiovert 135 microscope (Carl Zeiss AG, Germany). Two experiments were performed 

using the chromosome. Firstly, DOP-PCR (degenerate oligonucleotide primed 

polymerase chain reaction) was conducted using a 22-mer degenerated primer. 

Secondly, chromosome was digested using Sau3AI, Sua3AI specific adaptor was ligated 

to chromosomal DNA, and adaptor mediated PCR was conducted. Southern 

hybridization results showed that the amplified products were homogeneous with lily 

genomic DNA, indicating that DNA from the dissected chromosome has been 

successfully amplified by DOP-PCR or adaptor mediated PCR. The PCR products were 

cloned using TOPO TA cloning kit (Invitrogen, USA) to create a chromosome-specific 

library. Evaluation of 200 randomly selected clones showed that the size of the cloned 

inserts varied from 300 bp to 2000 bp. These results suggest that microdissection and 

microcloning of lily chromosome #1 is feasible. The approach used here could be 

applied to the genetic mapping and isolation of chromosome #1-specific genes which are 

conveying resistance against environmental stresses such as cold, drought, diseases so 

on. 


INTROGRESSION BREEDING IN GENUS 

TULIPA ANALYSED BY GISH 

Interspecific hybridization is an important tool in Tulipa breeding to introgress some 

important horticultural traits into new cultivars. In our study, the main goal is the 

introgression of resistance against Tulip Breaking Virus (TBV), which is found in some 

cultivars of Tulipa fosteriana (F) to T. gesneriana germplasm (G), the commercial 

assortment. The diploid (2n = 2x = 24) F 1 interspecific hybrids between T. gesneriana × 

T. fosteriana (GF hybrids) were backcrossed to T. gesneriana and a number of BC 1 

hybrids (GGF) have been analysed by genomic in situ hybridization (GISH) to evaluate 

the number of chromosomes derived from F and G genomes and the number of 

recombinant chromosomes. All of the BC 1 hybrids analysed were diploid (2n = 2x = 24). 

By GISH it was possible to distinguish chromosomes from both parental genomes as 

well as the recombinant chromosomes. Because the T. gesneriana parent was used for 

backcrossing, the number of G genome chromosomes (chromosomes which centromere 

was of T. gesneriana genome) predominated in the BC 1 progenies and varied from 

fourteen to eighteen whereas the total number of T. fosteriana chromosomes in hybrids 

ranged from six to ten. The number of recombinant chromosomes differed among 

hybrids from five to nine. For most genotypes there were two types of recombinant 

chromosomes. Those with a centromere of T. fosteriana chromosome with recombinant 

segment of T. gesneriana (F/G) and vice versa (G/F). Most recombinant chromosomes 

contained a combination of a single T. gesneriana and a single T. fosteriana fragment. 

The maximum number of recombinant segments per chromosome was two and their 

position ranged from distal to highly interstitial. For each genotype the percentages of 

each genome present in BC1 progenies was estimated. The percentages of T. fosteriana 

chromatin in hybrids ranged from 19.71 to 24.56%, while 25% was expected. 

LP7 

Agnieszka Marasek- 

Ciolakowska (1) 

M.S. Ramanna (2) 

Jaap M. van Tuyl (2) 


Pomology and Floriculture, 

Department of Physiology 

and Biochemistry, 

Pomologiczna Str. 18, 96- 

100 Skierniewice, Poland 

(2) Wageningen UR Plant 

Breeding,Wageningen 

University & Research 

Center, 

Droevendaalsesteeg 

1, 6708 PB Wageningen, 


agnieszkamarasek@wp.pl 


LP8 

R.K. Kishor 

G.J. Sharma 

Department of Life 

Sciences, Manipur 

University, Imphal-795003, 

India 

Medicinal Plants & 

Horticultural Resources 

Division, Institute of 

Bioresources & Sustainable 

Development, Imphal- 

795001, India 

gjs1951@rediffmail.com 

MORPHOLOGICAL AND MOLECULAR 

CHARACTERIZATION OF INTERGENERIC 

HYBRIDS BETWEEN THE ORCHID GENERA 

RENANTHERA AND VANDA 

The Indo-Burmese mega-biodiversity ‘hotspot’ houses a number of ornamental orchids 

belonging to rare, endangered, threatened and vulnerable species possessing highly 

attractive flowers. However, till date, many of these beautiful orchids growing in the 

pristine forests of north-east India have remained largely unexplored and unexploited. 

Renanthera imschootiana Rolfe is an IUCN red-listed rare and endangered orchid 

classified under Appendix-I of the Convention on International Trade in Endangered 

Species of Wild Fauna and Flora (CITES). This orchid is reknowned for its bright 

crimson, long-lasting floriferous spikes which bloom during spring every year. Vanda 

coerulea Griff. ex. L., Vanda testacea (L.) Reichb.f. and Vanda stangeana Reichb.f. 

(endemic to Manipur) are simultaneously deeply coloured flowers with high aesthetic 

values. Using Renanthera imschootiana as the female parent, we have crossed it with 

the three Vanda species and intergeneric hybrids have been synthesized, viz., 

Renantanda Kebisana Shija (R. imschootiana x V. coerulea), Renantanda Prof GJ 

Sharma (R. imschootiana x V. testacea) and Renantanda Momon Shija (R. imschootiana 

x V. stangeana). All these hybrids have been registered with the Royal Horticultural 

Society, London. We have also developed in vitro propagation protocols for these 

parents and their hybrids keeping in view of their conservational and eco-restorative 

considerations. The synthesized hybrids produced beautifully coloured flowers with 

shelf-lives lasting from 1-1 1 / 2 months. Morphological as well as molecular 

characterization of these hybrids have been made for confirmation of hybridity and other 

genetic information. 


DIVERSITY IN ROSA RUGOSA X ROSA 

HYBRIDA INTERSPECIFIC VARIETIES 

Modern roses represent a relatively narrow gene pool resulting in poor disease and 

abiotic stress resistance. Diploid Rosa rugosa have better disease resistance and higher 

tolerance for environmental stress conditions, however, their ornamental characteristics 

need improvement. Interspecific crosses between modern and Rugosa roses are difficult 

to obtain, but they could potentially yield resistant varieties with valuable ornamental 

properties. Eight Latvian varieties resulting from crosses with Rugosa type roses and six 

parents were analyzed with 21 microsatellite markers, to confirm the results of 

interspecific crosses and to identify presence of garden rose genetic material. Majority of 

varieties resulting from interspecific crosses showed strong presence of R. rugosa gene 

pool both in terms of genotype and phenotype. However, two crosses, ‘Abelzieds’ (R. 

rugosa ‘Alba’ x ‘Poulsen’s Pink’) and ‘Zaiga’ (R. rugosa ‘Plena’ x ‘Flammentanz’), 

exhibited presence of floribunda and climbing rose characteristics. Phenotypically only 

one interspecific variety ‘Zaiga’ showed garden type flower colour and leaf shape, but 

both cultivars could be used for breeding owing to good disease resistance, rigorous 

growth and ornamental characteristics. 

LP9 

Anta Sparinska (1) 

Romans Veveris (2) 

Renate Zarina (2) 

Nils Rostoks (2) 

Dzidra Rieksta (3) 

(1) Botanical Garden, 

University of Latvia, 2 

Kandavas Str., Riga, LV- 

1083, Latvia 

(2) Faculty of Biology, 

University of Latvia, 4 

Kronvalda Blvd., Riga, LV- 

1586, Latvia 

(3) National Botanical 

Garden, 1 Miera Str., 

Salaspils, Riga reg., LV- 

2169, Latvia 

anta.sparinska@lu.lv 


LP10 

W. Tera-arusiri (1) 

S. Mahadtanapuk (1) 

S. Anuntalabhochai (2) 

M. Sanguansermsri (3) 

W. Nanakorn (3) 

(1) School of Agriculture 

and Natural Resources, 

Naresuan University 

Phayao, Tumbol Maeka, 

Muang, Phayao, 56000, 

Thailand 

(2) Department of Biology, 

Faculty of Science, Chiang 

Mai University, Chiang Mai 

50200, Thailand 

(3) Naresuan University 



Thailand 

werachai_kig_bu@ 

hotmail.com 

BREEDING FOR RESISTANCE AND 

BIOCONTROL OF WILT DISEASE IN 

CURCUMA ALISMATIFOLIA GAGNEP BY 

BACILLUS SPP. 

The objectives of this study were to breed for resistance of Curcuma alismatifolia 

Gagnep to wilt disease caused by Rastonia species, by screening and collecting virulent 

isolates of Rastonia Moreover, in order to preserve the value of these ornamental 

flowers, and to reduce the dependence on toxic chemicals required to combat this 

disease the biocontrol potential of bacteria antagonistic to Rastonia, , was investigated. 

Over 500 bacterial strains, isolated from soil, leaf surfaces of C. alismatifolia Gagnep. 

and hot springs in the Chiang Mai province in Thailand, were screened in vitro for 

antagonistic activity against R. solanacerarum. Three isolates providing growth inhibition 

in vitro , were identified as Bacillus licheniformis, B.amyloliquefaciens and B. subtilis. 

Subsequently, these isolates were used for biocontrol in planta. One of the isolates, B. 

subtilis was shown inhibition greater than 70% when compared with the control and 

provided a statistically significant growth suppression of the wilt disease on the curcuma. 

Moreover, all of the antagonistic bacteria isolates were selected to against Anthracnose 

caused by Colletotrichum sp. in planta. The highest levels of the Anthracnose disease 

suppression occurred using B. licheniformis and B. subtilis on curcuma flower. The 

present results indicate that B. licheniformis, B.amyloliquefaciens and B. subtilis could be 

used to inoculate and reduce the symptoms of disease on Curcuma alismatifolia 

Gagnep. 


CURRENT STRATEGİES AND FUTURE 

PROSPECTS OF RESİSTANCE BREEDİNG İN 

ORNAMENTALS 

Ornamental crops pose several problems to breeding for disease and pest resistance. 

The large number of ornamental crops and the short turnover time of varieties limit the 

input invested to the individual variety. In addition many ornamental crops are polyploids 

hampering genetic analyses of resistance traits. In current breeding programms for most 

crops stringent selection for disease resistance is either omitted or performed at 

relatively late stages of the selection program. However, knowledge about the 

pathosystem including the genetic composition of both the host plant and the pathogen 

populations will allow efficient early selection for resistance in conventional breeding 

programs. These strategies can be significantly improved by the application of molecular 

diagnostic tools as e.g. molecular markers. Examples of improved selection schemes as 

well as marker applications will be shown for the pathosystem rose/blackspot. A limited 

number of ornamental crops are amenable to positional cloning (as e.g. roses) or 

Transposon tagging (as e.g. petunias) of genes allowing the de novo isolation of genetic 

factors important for disease resistance. Mid- and long term improvements can also be 

expected from current genome projects and the application of biotechnology. The 

identification of key factors with central functions in disease resistance via sequence 

homology and microsynteny will get easier with every completely sequenced genome. 

Here I present examples on how this information may be utilised in ornamental 

resistance breeding by using non transgenic strategies as for example Tilling and 

Ecotilling or by generating transgenic plants. 

L15 

Thomas Debener 

Leibniz University 

Hannover, Institute for 

Plant Genetics, 

Herrenhäuser Str. 2, D 

30419 Hannover, Germany, 

debener@genetik.unihannover.de 

Session Resistance breeding 41

L16 

Arwa Shahin 

Paul Arens 

Sjaak van Heusden 

Jaap van Tuyl 

Wageningen UR, Plant 


6700 AJ Wageningen, 


Arwa.shahin@wur.nl 

CONVERSION OF MOLECULAR MARKERS 

LINKED TO FUSARIUM AND VIRUS 

RESISTANCE IN LILIUM 

Lilies are one of the most economically important monocot flower bulbs. They are mainly 

cultivated in the Netherlands with a bulb acreage of more than 4000 ha. However, lily 

bulb production faces some challenges such as being susceptible to diseases like 

Fusarium oxysporum and Lily mottle Virus (LMoV). These two are the most important 

pathogens that cause serious damage to lily. Fortunately, some Asiatic lily hybrids 

showed high level of resistance to Fusarium and LMoV. The incentive for breeding 

resistances into the lily assortment is limited due to the relatively long juvenile phase (2-3 

years) and many selection’s cycles of breeding needed to place these desirable 

agronomic traits from different resistant parents into specific commercial cultivars. 

Therefore, using marker-assisted selection (MAS) could speed up the breeding process 

considerably. A genetic map of an intraspecific Asiatic backcross [Orlito (Pirate x 

Connecticut King) crossed with Connecticut King] was constructed using three different 

molecular marker systems (DArT, AFLP and NBS profiling). The disease tests for the 

two pathogens were carried out for four years on this Asiatic population [100 BC1].Four 

QTLs for Fusarium oxysporum and one for LMoV were localized on the map. The most 

tightly linked markers to the resistance will be converted into more robust PCR markers 

and use for breeding purposes. The most significant Fusarium’s QTLs were successfully 

converted and specific primers for Fusarium resistance are developed. 

42 Session Resistance breeding

SPECIFIC MAPPING OF DISEASE 

RESISTANCE GENES IN TETRAPLOID CUT 

ROSES 

Control of fungal diseases is a major constraint of cut-rose cultivation in greenhouses 

and in transportation around the world. Therefore, development of resistant cultivars is a 

promising way to reduce the use of chemicals required for controlling the diseases. 

Genetic analyses and breeding for resistance, however, are hampered by a high degree 

of heterozygosity and the polyploid nature of cultivated roses. 

Nucleotide-binding site (NBS) profiling of Van der Linden et al. (2004) was used as a 

tool enabling a more directed way of studying the genetics of resistance to pathogens 

responsible for diseases such as botrytis and powdery mildew. 

NBS profiling is a multiplex screening technique, producing amplified resistance gene 

analogue (RGA) fragments by using degenerated primers based on the conserved 

motifs present in the NBS domain of resistance genes. Since NBS regions are 

abundantly distributed and highly polymorphic within the plant genome, they are very 

suitable as markers to identify resistance genes. 

Twelve NBS degenerated primer/restriction enzyme combinations were used to 

genotype the whole rose tetraploid K5 population and its parents (Yan, 2005). To 

generate RGA profiles, the restriction enzymes: AluI, HaeIII, MseI, and RsaI were 

combined with primers NBS1, NBS3, and NBS5a6. The profiles were dominantly scored 

resulting in 135 polymorphic RGA markers which segregated in a 1:1 or 3:1 ratio. 

The set of 135 markers, representing uni- and bi-parental simplex markers, were 

mapped on the two available parental AFLP/SSR K5 maps with Joinmap 4.0 

(unpublished). This resulted in two parental maps of 1150 cM and 1160 cM with 203 

markers and 198 markers, respectively. The tetraploid maps will be used to dissect the 

genetic variation for resistance to powdery mildew resistance. 

Moreover, Rosaceae SSRs mentioned in the literature are currently tested on the K5 

population to obtain allelic bridges between the tetraploid and diploid genetic maps in 

rose and related species in order to align them. These bridges will improve cross-ploidy 

comparisons in roses in order to strengthen cut rose breeding. 

L17 

C.F.S. Koning- 

Boucoiran 

O. Dolstra 

C.G. van der Linden 

J. van der Schoot 

V.W. Gitonga 

K. Verlinden 

F.A. Krens 

Wageningen UR Plant 

Breeding. B.O. Box 16, 

6700 AA Wageningen, 


Carole.boucoiran@wur.nl 

Van der Linden C. G. et al. Efficient targeting of plant disease resistance loci using NBS 

profiling. Theoretical and Applied Genetics, 109:384-393. 

Yan Z. 2005. Towards efficient improvement of greenhouse grown roses: genetic 

analysis of vigour and powdery mildew resistance. PhD Thesis, Wageningen-UR, The 

Netherlands. 90pp.. 

Session Short presentations – Resistance breeding 43

L18 

A. Balode 

Institute of 

Agrobiotechnology, Faculty 

of Agriculture 

Latvia University of 

Agriculture, Liela 2, 

Jelgava, LV-3001, Latvia 

antra@ram.lv 

BREEDING FOR RESISTANCE AGAINST 

BOTRYTIS IN LILY 

The fungus Botrytis occurs worldwide and destroys various crops. In lilies, the strain 

Botrytis elliptica destroys leaves, flower buds and flowers. Reddish-brown spots, the first 

evidence of the disease, appear on the leaves. During wet weather, the spots eventually 

coalesce and the whole leaf collapses and decays. To avoid the use of chemical 

pesticides and to make growing of plants economically viable and ecologically safe, in 

lily breeding the current activities are directed towards the development of diseaseresistant 

cultivars. Crosses between cultivars with different levels of resistance to 

Botrytis elliptica (study year 2002) have been obtained. Estimation of parent plants and 

the produced hybrids have been done, by two years of testing, on plants grown in natural 

environment (2004-2005). The level of Botrytis resistance was rated visually from 0-4 

grades (0=healthy–4=very susceptible). When Botrytis-resistant parents were crossed, 

the hybrids were characterized by degrees of resistance to the disease; when Botrytissusceptible 

cultivars were crossed with cultivars resistant to the disease, the hybrids 

were rated intermediate – their susceptibility to the disease was dominant. Under field 

conditions, 10 perspective hybrids have been tested (2006-2007) for resistance to the 

grey mould. After two years of testing, no significant differences have been found. 

44 Session Resistance breeding

A DYSFUNCTIONAL CYMMV MOVEMENT 

PROTEIN GENE CONFERS RESISTANCE TO 

CYMMV IN DENDROBIUM ORCHID 

A Cymbidium mosaic virus movement protein gene with a site-specific mutation (mut11) 

under control of a ubiquitin promoter was inserted using biolistics into 2 Dendrobium 

varieties with the intention of creating CymMV-resistant orchids. Presence of the 

transgene in regenerated plants of D.x Jaquelyn Thomas ‘Uniwai Mist’ and D.x Jaq – 

Hawaii ‘Uniwai Pearl’ was confirmed by PCR using genomic DNA, and mut11-positive 

plants were potted ex vitro. Forty-two transgenic plants and 4 non-transgenic controls at 

the 4 to 6 leaf stage were inoculated with a 1: 1000 dilution of CymMV obtained from 

infected orchids. Plants were analyzed for systemic infection using tissue blot 

immunoassay (TBIA). Seventeen plants from 6 independent transformations remained 

virus-free, whereas all control plants were infected with CymMV within 1 month. Further 

analysis by RT-PCR showed that the mut-11 messenger RNA was detectable in only 2 

of these 17 plants. All plants were challenged again with a second CymMV inoculation 

as above followed by TBIA analysis after 1 month. Thirteen of 17 plants remained free 

from virus. A third challenge of these plants with CymMV as above was followed by TBIA 

analysis at 1 week, 2 weeks, 1 month, 3 months, 6 months and 12 months after 

challenge. Results at 2 weeks post-inoculation showed that all 6 controls and 4 individual 

transgenic plants, including the RT-PCR-positive plants, became systemically infected 

with CymMV. Eight transgenic plants from both varieties remained free from CymMV 12 

months after the third challenge. Lack of detectable mut11 mRNA in these resistant lines 

suggests that a post transcriptional gene silencing (PTGS) mechanism may be 

conferring resistance to CymMV. 

L19 

Kullanart Obsuwan (1,2) 

David A. Hieber (2) 

Rasika G. Mudalige- 

Jayawickrama (2) 

Adelheid R. Kuehnle (2) 

(1) Biology Department, 

Faculty of Science, 

Silpakorn University, 


(2) Tropical Plant and Soil 

Sciences, University of 

Hawaii, Honolulu, HI, 

96822, USA 

kulanart@su.ac.th 

Session Resistance breeding 45

L20 

Judith Blokland 

Plantum NL, Gouda, the 

Netherlands 

j.blokland@plantum.nl 

CAN WE STILL TAKE THE BREEDER’S 

EXEMPTION FOR GRANTED? 

Since the introduction of protection of plant varieties by plant breeders rights, the 

breeders’ exemption has always been a crucial principle of our breeding industry. It 

means that breeders have the freedom to use without permission all existing varieties for 

further crossing and selection. This has lead to a flourishing industry in which many 

different companies have had the opportunity to start breeding activities and obtain a 

position on the market. 

However, some developments might change this “freedom to operate”. Within the 

Plant Variety Protection system the interpretation of Essentially Derived Varieties can 

limit the breeders’ exemption to a serious extend. Second there is the increase of 

patents in the plant sector. In Europe varieties cannot be patented as such, but specific 

traits can be patented. All varieties that contain a patented trait fall under the scope of 

the patent which means one cannot use these varieties freely for breeding. Thirdly there 

is the biodiversity legislation which may limit the possibility to use varieties from certain 

countries without first closing agreements. 

Discussions about these topics are mainly dealt with on a political or juridical level. 

Since these developments are likely to have a very big impact on breeding and research 

activities it is of great importance that also breeders enter into these discussions. In my 

presentation I will contemplate on the different positions that exist about the 

interpretation of the EDV-concept and about the effect of patents in the plant sector. 

46 Session Plant Breeder’s Rights

ESSENTIALLY DERIVED VARIETIES IN 

ORNAMENTALS 

In ornamentals, natural or induced mutants are a common phenomenon. Such mutants 

or ‘sports’ may obtain Plant Breeders’ Rights (PBR) when shown distinct from all existing 

varieties. To protect the interests of the breeder of the original variety the International 

Union for the Protection of New Varieties of Plants (UPOV) has introduced the concept 

of ‘essentially derived varieties’ (EDVs). In the UPOV 1991 act several ways of obtaining 

an EDV are described, mutation being one of them. When a variety is shown an EDV, 

authorisation for commercial exploitation is needed from the breeder of the initial variety. 

There is considerable debate ongoing about which approaches to use for determining 

essential derivation and also which thresholds should be used in the different plant 

species. For determining whether a variety should be considered essentially derived 

from an existing variety two conceptually different approaches can be taken. The first 

one is based on genetic conformity, the second on a forensic approach. For the 

implementation of the EDV concept using the conformity approach it is important that 

similarities between unrelated varieties can clearly be separated from essentially derived 

varieties. In the forensic approach the high genetic similarity between original variety and 

mutant is taken as a starting point. The basic idea is to calculate the probability that a 

second, putatively derived, variety would have a profile identical to the initial variety, 

given an independent breeding history. Both approaches will be illustrated and ways to 

implement the EDV concept discussed 

L21 

Ben Vosman 


Breeding, P.O.Box 16, 


Netherlands 

ben.vosman@wur.nl 

Session Plant Breeder’s Rights 47

L22 

M.J.M. Smulders 

D. Esselink 

R. E. Voorrips 

B. Vosman 

Plant Research 

International, P.O. Box 16, 

NL-6700 AA Wageningen, 



ANALYSIS OF A DATABASE OF DNA 

PROFILES OF 734 HYBRID TEA ROSE 

VARIETIES 

Rose is the largest ornamental crop. Over 25,000 varieties of modern roses have been 

described, of which more than 10,000 hybrid teas. Such large numbers of varieties may 

cause problems in the DUS testing context. A major problem for all countries carrying 

out DUS tests is the requirement to compare new varieties to all other varieties in 

common knowledge. Clearly, strict adherence to this concept is logistically and 

financially impossible in a species such as rose, which is cultivated around the world. 

Another problem are the reference collections. Maintaining a living reference collection is 

unattractive because of the high costs associated and disease problems. When no 

reference collections are maintained, reference varieties need to be obtained from the 

breeders. It is important that the DUS examination office can quickly verify the identity of 

the material submitted. For this aspect of quality assurance, molecular markers are 

ideally suited, as they are highly discriminating and can be assayed rapidly and relatively 

cheaply. 

DNA microsatellites (simple sequence repeats, SSRs) are highly polymorphic and 

have the advantage of providing a co-dominant marker system based on a PCR 

technology. When analyzed as sequenced-tagged microsatellite site (STMS) markers, 

they provide simple banding patterns that are easy to record and are especially suitable 

for automated and objective analysis. In addition, the resultant data can be readily stored 

in a database. New varieties or new markers can be easily added to an existing 

database. We used a set of 11 microsatellite markers developed for rose (Esselink et al., 

Theor Appl Genet 106: 277-286, 2003) to generate a database of molecular profiles of 

734 entries of Hybrid tea varieties, including all new varieties of the period 2000-2005. 

Here, we report on the analysis of the molecular data in detail. Specifically, we have 

looked at discriminative power of the markers, reproducibility of the results, genetic (sub) 

structure in the set of varieties analyzed, as well as correlation between molecular and 

DUS characteristics. 

48 Session Plant Breeder’s Rights

BACK TO BASICS FOR NEW CROP 

DEVELOPMENT 

Breeding of new ornamental crops is one of the most rewarding professions in the world. 

Yet researchers are also faced with challenges that require both new and innovative 

research, as well as the utilization of trusted classical breeding methods. Although new 

breeding methods like marker assisted selection and genetic modification opens up 

numerous possibilities, these methods can often not be applied to new (relatively 

unknown) crop development. First of all, there is a lack of basic information needed for 

application of these methods and secondly, the commercial value of these crops does 

not justify the use of such expensive methods. Breeders of new crops are, thus, often 

faced with a lack of basic information, requiring them to go “back to basics” and the 

utilization of more classical breeding strategies. The requirements for new crop 

development will be discussed on the basis of the experience acquired with the 

development of Lachenalia cultivars. When developing new crops, researchers work 

mostly with a number of species and the information required needs to be generated 

through researching market potential, genetic background and selection procedures. 

New crop development requires, first of all, the collection and characterization of 

germplasm, followed by the establishment of basic information on reproductive biology 

and market requirements, or the aim for breeding. Inter-specific hybridization and the 

establishment of specific selection criteria form the major part of breeding in new crops, 

and this is often complemented by the induction of polyploids. For the successful 

continuation of these processes it is necessary to establish a basic cytogenetic 

background for the crop. This becomes even more important when working with 

complex/diverse genera. The cytogenetic research can complement molecular 

systematic research to establish the relationships between the different species in the 

breeding program. The value of cytogenetic and molecular systematic studies in 

genetics and breeding are demonstrated through the research done on Lachenalia 

species. Mutation induction is another breeding strategy that requires basic background 

information for successful application. This includes the establishment of basic in vitro 

protocols and the selection of specific candidate plants. Only after the establishment of 

these basic information systems can the breeder move to advanced techniques. 

L23 

R. Kleynhans 

ARC-Roodeplaat Vegetable 

and Ornamental Plant 

Institute, Private bag X293, 

Pretoria, 0001, South Africa 

Rkleynhans@arc.agric.za 

Session Breeding and Genetics 49

L24 

Neil O. Anderson (1) 

Adnan Younis (2) 

Ellie Opitz (1,3) 

(1) Dept. of Horticultural 

Science, University Of 

Minnesota, St. Paul, MN 

55108 U.S.A. 

(2) Institute of Horticultural 


Agriculture, Faisalabad, 

38040 Pakistan 

(3) Dept. of Animal 

Science, University Of 

Minnesota, St. Paul, MN 

55108 U.S.A 

ander044@umn.edu 

DEVELOPMENT OF COLOURED, NON- 

VERNALIZATION-REQUIRING SEED 

PROPAGATED LILIES 

Recent discovery of seed-propagated Lilium x formolongi hybrids that flower 

continuously in 1 dominant 

VER1, VER2 alleles) with frost-tolerance, day neutrality, and winter hardiness is an 

unprecedented combination in Lilium. Our objective to obtain coloured L. x formolongi 

was to use Class I lilies that initiate flower buds prior to a cold treatment as parents, e.g. 

Lilium martagon. Reciprocal interspecific crosses between fertile parents (1-L. x 

formolongi, 7-coloured L. martagon) were made in both directions to generate 

segregating hybrid (F 1 ) and inbred (F 2 ) progenies. A total of 8,826 F 1 seeds or embryos 

were generated. Embryo rescue was employed when L. martagon was the female, 

although viable embryos never germinated after ~1 yr. in culture. Mean germination 

ranged from 0% - 1.02% for in situ ripened seed. Hybridity for one successful cross (07L- 

14; L. x formolongi [00L-111-343 x 51-202-1] x L. martagon ‘Cadense’) was analyzed 

using morphological (flowering, leaf & internode number, leaf length:width ratios, 

compatibility, no. flower stalks & flowers, rosetting) and molecular markers (ISSR 

primers). Twenty-two 07L-14 genotypes were genetically similar to the female parent 

whereas 86 aligned closely with the male (‘Cadense’). The number of shoots/plant was 

the only quantitative trait co-segregating with ISSRs. Flowering traits were intermediate 

to, but significantly different than, either parent. A majority of the plant height and 

inflorescence lengths were significantly greater than the parents. Two F 1 s had slight 

flower colouration in the petals and tepals; one hybrid had an open-faced rather than 

trumpet flower (similar to an Oriental type). 

50 Session Breeding and Genetics

GROWTH AND DEVELOPMENT OF CUT ROSE 

CLONES; INDIRECT SELECTION FOR YIELD 

The shoot yield of 177 large-flowered own-rooted cut rose genotypes, grown in a heated 

glasshouse, was recorded for 40 weeks in year # 1. These genotypes showed a normal 

distribution over 8 yield classes : 420 shoots plant -1 . 5 Genotypes from each 

yield class (total of 40 genotypes) were propagated as cuttings and planted in the 

glasshouse early year # 2. The growth and development of these young plants were 

studied until each plant had 2 bottom-breaks (Bb). The mean number of days to the 

appearance of the 1 st and the 2 nd Bb, were 43.1 and 57.1 days after planting; days to 

flowering of these shoots took 35.5 and 34.2 days from their appearance; at flowering 

the shoot lengths were 77.5 and 83.4 cm; mean length increases were 23 and 26 

mm.day -1 . Highly significant negative correlation (r = - 0.69 +++ ) between the shoot yield in 

year # 1, and the number of days to appearance of the 1 st Bb of the same genotypes in 

year # 2, shows unique possibilities to indirect selection for future yield of clonal plants. 

The probable role of endogenous hormone action in the carbon partitioning in rose 

plants is discussed. 

L25 

D.P. de Vries 1) 

L.A.M. Dubois (2) 

(1) Foundation Sub Rosa, 

P.O.B. 4097, 6710 EB Ede, 



International (PRI), P.O.B. 

16, 6700 AA Wageningen, 


devriesrosa@hetnet.nl 


L26 

Kell Kristiansen 

Karen K. Petersen 

Institute of Horticulture, 

Faculty of Agricultural 

Sciences, Aarhus 

University, Kirstinebjergvej 

10, DK-5792 Aarslev, 

Denmark 

kell.kristiansen@agrsci.dk 

IN VITRO MUTAGENESIS OF ASTER NOVI- 

BELGII 

Aster novi-belgii is an important ornamental in the Danish pot plant industry with 

approximately 7 mill. pots produced annually in Denmark. The main commercial cultivars 

have been bred by Institute of Horticulture and are owned by the Danish Aster 

Association. During 15 years of conventional crossings more than 30 cultivars in two 

series (Viking and Victoria) have been granted plant breeders rights. The cultivars have 

since dominated the pot asters market worldwide. To increase breeding efficiency and 

explore new possibilities mutagenesis were investigated in two cultivars (‘Fanny’ and 

‘Jane’) of the Victoria series. An in vitro culture system based on adventitious shoot 

formation was developed for a number of cultivars to reduce chimerism. The in vitro 

culture system used leaf blade sections from in vitro grown plantlets as explant source 

and adventitious shoots were produced within eight weeks on a MS based medium. 

To generate mutants shoots from in vitro grown plants were irradiated with gamma 

rays prior to induction of adventitious shoots. The highest amount of radiation allowing 

adventitious shoot formation was initially determined to 18 Gy. Plants from the two 

cultivars was thereafter produced and evaluated in the greenhouse for mutants. Special 

interest was placed on flower characteristics, ie. colour, size and whorls of ray florets. 

The two cultivars differed with respect to mutations with ‘Fanny’ giving a higher number 

and more interesting mutants. Flower colours ranged from pink, to purple and dark violet. 

The number of whorls of ray florets in capitula varied from a single to numerous and 

plants only producing ray florets were found. Further variation in plant height, branching 

ability and time to flowering was observed among the regenerants. Despite adventitious 

shoot formation plants showing chimerism were found. 

In vitro mutagenesis of Aster novi-belgii is a promising tool to improve the 

assortment; however, the potential of this technique depends on the cultivar used for 

mutagenic treatment and chimerism cannot be totally avoided. 


BREEDING THE TOMATO MICRO-TOM MODEL 

SYSTEM FOR ORNAMENTAL VALUE 

Taking advantage of its small size (8 cm tall) and short life cycle (70 days), the 

ornamental tomato cv Micro-Tom (MT) had been proposed as a model system for 

tomato genetics. Ever since, MT are being increased used for large scale mutagenesis 

and transgenic plant production, as well as the introgression of well characterized 

mutations and allelic variation already known in other cultivars and tomato wild species. 

Using such approaches we noticed that many mutations obtained in the MT background 

could be used to improve its value as an ornamental. Here we report the introgression of 

various mutations affecting fruit colour and morphology into the MT background. The 

introgression process consisted of series of crosses and backcrosses. In each cross, 

pollen was collected of the parent plants and used to fertilize emasculated MT flowers. 

The first cross F1 was selfed to obtain a recombinant F2 which was selected for small 

size and the mutation of interest. The selected plants were backcrossed with MT up to 

the sixth generation (BC6), with selfing every two generations to screen for homozygous 

recessive mutants. Plants were grown in 150 ml pots containing a 1:1 mixture of 

commercial pot mix and expanded vermiculite, supplemented with 1 g l -1 10:10:10 NPK 

and 4 g l -1 lime in a greenhouse with mean temperature of 28 o C, 11.5 h/13 h 

(winter/summer) photoperiod, and 250 to 350 µmol photons m -2 sec -1 PAR irradiance 

obtained by reduction of natural radiation with a reflecting mesh. Different true type 

genotypes combining reduced size and fruit colour variation were obtained upon 

introgression of the mutations Del, old gold (og), Beta (B), green stripe (gs), pink fruit (y), 

yellow flesh (r), tangerine (t), apricot (at) and green flesh (gf) in the MT background. Four 

mutations affecting fruit format, i. e. ovate (o), fascinated (f), sun and fs8.1, were also 

introgressed in the MT. These mutations and the combination of them greatly extended 

the value of MT as an ornamental and introduced the possibility to explore the large 

diversity of tomato genetic for these propose. 

L27 

Lázaro Eustáquio 

Pereira Peres 

Universidade de São Paulo 

(USP), Escola Superior de 

Agricultura ¨Luiz de 

Queiroz¨ (ESALQ), 

Departamento de Ciência 

Biológicas (LCB), Av. 

Pádua Dias, 11 CP. 09, 

CEP 13418-900, Piracicaba 

– São Paulo, Brazil 

lazaropp@eslaq.usp.br 


L28 

L. Leus 

K. Van Laere 

A. Dewitte 

J. Van Huylenbroeck 

ILVO, Plant Sciences Unit, 

Applied genetics and 

breeding, Caritasstraat 21, 


leen.leus@ilvo. 

vlaanderen.be 

FLOW CYTOMETRY FOR PLANT BREEDING 

Flow cytometry is a well known and established technique for ploidy analysis in plant 

breeding. In general it is used to characterize parent plants, to screen offspring after 

interploidy crosses, to control ploidy levels during seed multiplication or to evaluate 

haploidisation or polyploidisation experiments. Recent developments in the apparatus 

itself as well as in automated sample preparation simplified the technique and made it 

more cost effective. This allows breeders to have an easier access and to apply the 

technique in their breeding programs. 

Commonly for ploidy analysis DAPI staining is used to measure the relative DNA 

content in the cell. DNA content is then correlated to chromosome amounts. However no 

information is obtained on exact chromosome numbers. By flow cytometry more specific 

information linked to genome sizes can be obtained. Examples will be given on the use 

of genome sizes to chose parent plants for interspecific hybridization and to evaluate the 

hybrid nature of seedlings within Buddleja and Hydrangea. Besides flow cytometry as a 

tool to recognize unreduced pollen in Begonia and Hibiscus and to detect interesting 

modifications like e.g. sports in Rhododendron will be discussed. 


A PROTOCOL FOR HIGH RATE AGRO- 

BACTERIUM-MEDIATED TRANSFORMATION 

OF LILIUM 

Lilies are economically important, mainly because of their large and attractive flowers. 

Low rate of transformation in Lilium is the main barrier for molecular breeding of this 

important plant. In this study, an efficient system for Agrobacterium mediated 

transformation of Lilium × formolongi and Acapulco was established using mersitematic 

nodular calli. The calli were inoculated with A. tumefaciens strain EHA101 containing the 

plasmid pIG121-Hm which harbors intron-containing β-glucuronidase (GUS) gene under 

the control of a 35S cauliflower mosaic virus promoter, hygromycin phosphotransferase 

(HPT) gene, and neomycin phosphotransfease (NPTII) gene as reporter genes. The 

effects of different concentration of calcium (0, 11, 22, 44 and 88 mg/l), different 

carbohydrates (sucrose, glucose and arabinose) and two types of MS media (MS full 

strength and MS modified) in inoculation and co-cultivation medium were considered. In 

all treatments, 10mM MES and 100 µM acetosyringone were used. In medium without 

calcium the rate of transformation slightly increased. Interestingly, the use of MS 

modified medium (without some macro elements) containing sucrose dramatically 

increased the transformation efficiency. The highest rates of transformation 25% and 

23% were attained using MS modified medium containing sucrose in Lilium × formolongi 

and Acapulco, respectively. The hygromycin resistance calli were successfully 

regenerated into plantlets on hormone free MS medium. Transgenic plants were 

confirmed by GUS histochemical assay, PCR and Southern blot analyses. 

LP11 

P. Azadi 

D.P. Chin 

K. Kuroda 

R.S. Khan 

M. Mii 

Laboratory of Plant Cell 

Technology, Faculty of 



Matsudo City, Chiba 271- 

8510, Japan 

azadip22@yahoo.com 


LP12 

B. Christensen (1) 

S. Sriskandarajah (2) 

E.B. Jensen (3) 

R. Müller (3) 

(1) Institute for Agri 

Technology and Food 

Innovation, Højbakkegård 

Allé 21, DK-2630 Taastrup, 

Denmark 

(2) Department of Plant 

Biology and Forest 

Genetics, Uppsala 

Biocenter, Box 7080, SE- 

750 07, Uppsala, Sweden 

(3) University of 

Copenhagen, Faculty of 

Life Sciences, Department 

of Agricultural Sciences, 

Crop Science, 

Højbakkegård Allé 21, DK- 

2630 Taastrup, Denmark 

ren@life.ku.dk 

TRANSFORMATION WITH ROL-GENES OF 

AGROBACTERIUM RHIZOGENES AS A 

STRATEGY TO BREED COMPACT 

ORNAMENTAL PLANTS WITH IMPROVED 

POSTHARVEST QUALITY 

The aim of the present investigation was to find and evaluate an alternative strategy to 

produce potted plants without using chemical growth regulators by transformation with 

Agrobacterium rhizogenes. In Kalanchoe blossfeldiana, root inducing (Ri) lines were 

regenerated from hairy roots produced by transformation with the natural occurring 

bacteria A. rhizogenes. Transformed plants displayed distinct changes in plant 

morphology,.A number of Ri-lines were analysed thoroughly in greenhouse trials and 

characterisation showed that both growth and development were affected. Several lines 

were compact and exhibited an increased number of lateral shoots. Time to flowering 

was the same as in control plants in one of the investigated lines and delayed by only 

three days in another Ri-line; other lines showed delayed flowering compared to control 

plants. 

In ethylene free environment, transformants performed better than control plants and 

single flowers lasted longer. In response to exogenous ethylene, flowers of Ri-lines 

exhibited tolerance while chemically growth regulated and control plants were sensitive. 

Possible mechanisms behind the improved postharvest performance of plants 

transformed with rol-genes are presently investigated. 

Crossing with established lines documented heredity of the traits of interest since 

dwarfism was obtained in the offspring. The presence of rol-genes was proved in the 

progeny exhibiting dwarfism. The heredity of the traits will be further investigated. A 

compact plant without delayed flowering, and improved postharvest performance is 

valuable for further breeding programmes. 


NEW GENOTYPES OF HIBISCUS ROSA- 

SINENSIS THROUGH CLASSICAL BREEDING 

AND GENETIC TRANSFORMATION 

Hibiscus is the largest genus of the mallow family (Malvaceae), a group comprising 116 

genera, many of which are economically valuable. Several Hibiscus species are used all 

over the world as ornamental plants. The most popular one, H. rosa-sinensis shows a 

high variety of flower colours and shape. However, its use as an ornamental plant is 

currently restricted by a few constraints: 1) most of the commercialised pot-plant 

varieties are obtained by applying growth retardants; 2) the range of colours and types in 

use is limited if compared with the available ones; 3) the present cultivars are not fit for 

the Mediterranean region in terms of propagation and growth rates, as well as flowering; 

4) the market requires frequent introductions of novelties. 

In 2006, a breeding programme was established at CRA-FSO with the aim of 

selecting specific cultivars suitable for pot-plant production and creating new genotypes 

well-adapted to the Mediterranean climate. A partial diallelic cross design (reciprocals 

without self-fecundations) was used to test the ability of the collected cultivars to produce 

valuable progenies. Only a limited number of cross progenies resulted to combine 

desirable characters with a reasonable degree of fertility. A range of very good female 

parents able to produce seeds in all cross combinations was found. The first selected 

cultivars are presented. 

At the same time, another approach was followed, in order to investigate whether 

desirable morphological modifications (plant size reduction for pot plant cultivation) could 

be obtained by transforming H. rosa sinensis with Agrobacterium rhizogenes. In vitro 

seedlings were used as sources of explants for the transformation experiments. Two A. 

rhizogenes strains (ATCC 15834 and NCPPB 1855) were employed. Axenic hairy root 

cultures were established about 4 months after inoculation. Hairy roots grew vigorously 

on hormone-free medium whereas normal roots did not. Transformed roots displayed a 

typical hairy root phenotype characterized by fast growth, high lateral branching and lack 

of geotropism. So far, after more than one year of cultivation, a clone of hairy root 

deriving from a cotyledon formed a friable yellowish callus at root node level; several 

adventitious buds are spontaneously regenerating from it. Work is still in progress. 

LP13 

Luca Braglia 

Antonio Mercuri 

Laura De Benedetti 

Marco Ballardini 

Cesare Bianchini 

CRA-FSO Research Unit 

for Floriculture and 

Ornamental plants, Corso 

degli Inglesi 508, 18038 

Sanremo – ITALY 

a.mercuri@istflori.it 

antonio.mercuri@entecra.it 


LP14 

Youn-Hwa Joung (1) 

Jong-Cheol Choi (1) 

Won-Hee Kim (2) 

Gwang-Yeon Gi (3) 

Tae-Ho Han (1) 

(1) Dept. of Plant 

Biotechnology, Chonnam 

National University, 

Gwangju 500-757, Korea 

wageningen@hanmail.net 

(2) National Horticultural 

Research Institute, RDA, 

Suwon 440-760, Korea 

(3) Jeollanamdo 

Agricultural Research and 

Extension Servies, Naju 

520-830, Korea 

INVESTIGATION OF THE FACTORS 

AFFECTING CROSS-FERTILIZATION RATE IN 

ROSE 

Rose has most long history of artificial improvement by man-kind in ornamental plants. 

Due to their complex genetic constitution, rose is heavily self-incompatible and gives in 

low cross-fertilization rate by crossing genetically distant pairs. Consequently, low crossfertilization 

rate results in a negative effect on breeding process. In this study, we 

investigated the factors that would influence cross-fertilization rate of roses. Correlation 

analysis was performed between cross-fertilization rate and genetic distances of their 

parents. Analysis of variance was also performed to study the paternal and maternal 

effects for the cross-fertilization rate. Reliable data for cross-fertilization rate was 

obtained from Jeollanamdo Agricultural Research and Extension Servies, Korea in 2006. 

Thirty two cross combination provided cross-fertilization rate, and their genetic distances 

were obtained by using RAPD marker analysis. In RAPD analysis, we used 16 primers 

and obtained clear 101 RAPD markers. Genetic distances were obtained by using Nei’s 

coefficients. Relationship between cross-fertilization rate and genetic distance of parents 

was conducted by using correlation analysis. The correlation coefficient was as low as 

0.058 that was no significant. Consequently, we determined that overall genetic 

distances do not influence the cross-fertilization rate by any means. The paternal and 

maternal effects for the cross-fertilization rate were determined by. The result of analysis 

of variance showed that only maternal side influences the cross-fertilization rate. This 

guide that rose breeder must have desirable materials that can be served as receiver 

plants. Moreover, we expect more complicated factors will influence on the crossfertilization 

rate and more approaches should envision on this matter. 


CLONING OF THE ACC SYNTHASE GENE 

FROM CURCUMA ALISMATIFOLIA GAGNEP 

AND ITS USE IN TRANSFORMATION STUDIES 

The goal of this work was to suppress the expression of the ACC synthase gene in the 

Siam tulip, Curcuma alismatifolia Gagnep. A cDNA fragment encoding ACC synthase 

from C. alismatifolia Gagnep. was isolated and expression studies were done. To isolate 

this gene a pair of primers was designed from the highly conserved motif of ACC 

synthases in various plant species. The PCR product of 600 bp. in C. alismatifolia 

Gagnep. was subcloned into the pGEM-T easy vector resulting in pCa-ACSI. After 

sequencing, the sequence and its deduced amino acid sequence were highly 

homologous to the ACC synthases from other plants. To determine expression patterns 

of pCa-ACSI, a Northern blot analysis showed that the expression of the pCa-ACSI gene 

was in the bracts of curcuma, the highest expression was observed 2 days after cutting 

the flowers. Ca-ACSI was subcloned in pBI121 resulting in pBI121-Ca-ACSI and 

transformed into leaf tissues of Torenia foumieri and retarded shoots of C. alismatifolia 

Gagnep via Agrobacterium tumefaciens strain AGLO. Putative transformants, with the 

gene in an antisense orientation, were investigated by PCR analysis, GUS assays and 

Southern blotting. The transgenic plantlets were transferred to pots containing soil for 

cultivation in growth chamber. At present, the transgenic plants grow happily in the 

greenhouse and their phenotypic is under investigation. 

L15 

S. Mahadtanapuk (1) 

M. Sanguansermsri (2) 

T. Handa (3) 

W. Nanakorn (2) 

S.Anuntalabhochai (4) 






Thailand 

(2) Naresuan University 



Thailand 

(3) Graduate School of Life 

and Environmental 


Tsukuba, Tsukuba, Ibaraki 

3058572, Japan 


Faculty of Science, Chiang 

Mai University, Chiang Mai 


burinka@hotmail.com 

Session Short presentation 59

LP16 

T. Basaki (1) 

M. Jafarkhani Kermani 

(1) 

S.M. Pirseyedi (1) 

M.R. Ghaffari (1) 

A. Haghnazari (2) 

P. Salehi Shanjani (3) 

P. Koobaz (1) 

M. Mardi (1) 

(1) Agricultural 

Biotechnology Research 

Institute of Iran (ABRII), 

Mahdasht Road, Karaj, Iran 

(2) Zanjan University, 

Zanjan, Iran 


Forests and Rangelands, 

P.O.Box 13185-116, 

Tehran, Iran 

mardi@abrii.ac.ir 

ASSESSING ROSA PERSICA GENETIC 

DIVERSITY USING AMPLIFIED FRAGMENT 

LENGTH POLYMORPHISMS ANALYSIS 

The genetic diversity among 128 Iranian Rosa persica (R. persica) accessions in the 

different populations was analyzed. Amplified fragment length polymorphisms (AFLP) 

technique was used to produce 171 polymorphic fragments. The number of polymorphic 

loci ranged from 101 to 147 and the polymorphism information content (PIC) varied from 

0.289 to 0.073, with an average of 0.16. This shows extreme variability and genetic 

diversity among the studied R. persica populations. An indirect estimate of the number of 

migrants per generation (Nm=0.376) indicated that gene flow was relatively low among 

populations of the species. Cluster analysis using the UPGMA method grouped all 

accessions into six clusters. The results did not show relative agreement with the 

genotypes’ region of origin. Based on an analysis of molecular variance, 48% of the 

genetic variation of R. persica was within population and 52% was among populations. 

The present analysis revealed that Iranian R. persica genotypes are highly variable and 

genetically distinct from their origins. The apparent unique nature of the R. persica 

genotypes revealed by our results supports the case for the implementation of more 

intense characterization and conservation strategies, and provides useful information to 

address breeding programmes and germplasm resource management in Rosa spp. 


REDESIGNING FLORAL ARCHITECTURE: 

EFFICIENT MODIFICATION OF AGRONOMIC 

TRAITS BY CRES-T 

LP17 

While many of the genetically-modified plants now benefiting our daily life, high costs for 

their development and commodification is still a major problem. One effective way to 

solve this problem is to increase productivity of useful transgenic plants by targeting 

transcription factors, because most of the known plant mutant phenotypes have been 

shown to be caused by the disruption of their function. From this perspective, we applied 

Chimeric REpressor gene-Silencing Technology (CRES-T) to various kinds of 

ornamental flowers. CRES-T is an efficient gene silencing system in which the chimeric 

repressors derived from various transcription factors dominantly suppress the expression 

of the respective target genes, and the resultant transgenic plants exhibited loss-offunction 

phenotypes specific for the transcription factors even in the presence of 

redundant transcription factors. More than 100 Arabidopsis-derived chimeric repressors 

driven by CaMV35S promoter were individually or collectively introduced into 

chrysanthemum, torenia, cyclamen, morning glory, lisianthus and gentian to evaluate the 

availability and general versatility of CRES-T for improving floral traits. We found that 

CRES-T functions in all the species tested, even in the hexaploid chrysanthemum. It is 

useful not only for producing novel petal colour patterns and shapes as well as leaf 

shapes, but also for controlling agronomic characters such as ethylene sensitivity or 

flowering regulation. In addition, we have succeeded to revive a lost garden variety of 

morning glory which only remained in some ancient texts. Now we are improving and 

applying this system also to major flowers such as roses and carnations to improve the 

commercial value. In this presentation, we show some latest data on the CRES-Tapplied 

transgenic flowers and discuss how we should approach and handle this new 

technology. Some information including graphical contents of transgenic plant 

phenotypes is available to the public in our database “FioreDB” . This work was supported by the Program for Promotion of Basic 

and Applied Researches for Innovations in Bio-oriented Industry. 

Norihiro Ohtsubo (1) 

Takako Narumi (1,2) 

Katsutomo Sasaki (1) 

Hiroyasu Yamaguchi (1) 

Masahito Shikata (1) 

Nobutaka Mitsuda (3) 

Tomotusgu Koyama (3) 

Yoshimi Umemura (3) 

Miho Ikeda (3) 

Kyoko Matsui (3) 

Keiichiro Hiratsu (3,4) 

Kanji Isuzugawa (5,6) 

Reiko Endo (5) 

Kazuo Ikeda (5,7,8) 

Tomoyuki Sakai (5) 

Kumi Saito (5) 

Michiyuki Ono (9) 

Masahiro Nishihara (10) 

Takashi Nakatsuka (10) 

Teruhiko Terakawa (11) 

Yoshikazu Tanaka (12) 

Masaru Ohme-Takagi (3) 

Ryutaro Aida (1) 

(1) Natl. Inst. Floricultural Sci., 

NARO, Tsukuba, Ibaraki 305- 

8519, Japan; (2) Fac. Agric., 

Kagawa Univ., Miki-cho, Kagawa 

761-0795, Japan; (3) Res. Inst. 

Genome-based Biofac., AIST, 

Tsukuba, Ibaraki 305-8562, 

Japan; (4) Dept. Appl. Chem., 

Natl. Defense Acad. Japan, 

Yokosuka, Kanagawa 239-8686, 

Japan; (5) Yamagata General 

Agric. Res. Center, Dept. Agro- 

Produc. Sci., Sagae 991-0043, 

Japan; (6) Yamagata Pref. Gov. 

Office, Yamagata 990-8570, 

Japan; (7) A Non-Profit Corp. 

Yamagata Pref. Org. for Prom. of 

Agric., Sagae 991-0043, Japan; 

(8) Tottori Univ., Tottori 680- 

8533, Japan; (9) Grad. School of 

Life and Environmental Sci., 

Univ. of Tsukuba, Ibaraki 305- 

8572, Japan; (10) Iwate Biotech. 

Res. Center, Kitakami, Iwate 

024-0003, Japan; (11) Hokko 

Chem. Ind. Co., Atsugi, 

Kanagawa 243-0023, Japan; 

(12) Suntory Ltd. Inst. Plant Sci., 

Shimamoto-cho, Mishima-gun, 

Osaka 618-8503, Japan. 

nohtsubo@affrc.go.jp 


LP18 

Cristina Regis 

Marina Laura 

Cristina Borghi 

Andrea Allavena 

CRA – Unità di Ricerca per 

la Floricoltura e le Specie 

Ornamentali, Corso Inglesi 

508, 18038 Sanremo (IM), 

Italy 

a.allavena@istflori.it 

KALANCHOE X HOUGHTONII: SSH AND 

MICROARRAY ANALYSIS TO SCREEN GENES 

INVOLVED IN VIVIPARY 

Vivipary, referred here as the formation of novel complete plantlets on mature organs, 

has been reported in many families as an asexual propagation strategy. In K. 

xhoughtonii (Crassulaceae), viviparous plantlets are formed on leaf margin notches in 

response to a long day photoperiod and their appearance follow a basipetal fashion. To 

identify genes involved in this process, suppression subtractive hybridisation libraries 

(SSH) were prepared. 200 c-DNA clones were classified and grouped into fourteen 

functional categories according to Goldberg database 

(http://estdb.biology.ucla.edu/PcEST/). 

630 sequences (200 SSH library, 48 database Kalanchoe genus, 382 other species 

database genes) were used as probes for microarray analysis according to the 

CombiMatrix technology. A 4x2K Custom Array TM was synthesized . 

RNA was extracted from margin of leaves at 7 stage of development before buds 

emission (5mm to 50 mm) during long-day photoperiod (permissive conditions) and one 

stage during short-day photoperiod (non permissive condition). Three replications for 

each sample were prepared. From double strand cDNAs antisense RNAs (a RNA) were 

synthesized and amino-allil-UTP incorporated and coupled with Alexa Fluor ® 647. 

Microarray was hybridated according to CombiMatrix protocol. Data were extracted with 

CombiMatrix Microarray Imager software and exported into Microsoft Excel for 

computing of mean, median and standard deviation. Person's correlation was computed 

and data normalized. After background removal, probes were reduced to 484. “Fold 

change” method (FC=2) was used to compare levels of gene expression of samples in 

permissive conditions vs sample in non permissive condition. Significance Analysis of 

Microarrays Statistic (SAM method) generated 263 significant modulated genes with 

FDR of 5%. 


POLLEN CHARACTERISTICS AFFECT SEED 

PRODUCTION OF ROSE CULTIVARS 

Seed production and germination rates are important bottlenecks in rose breeding. 

Most cut roses are Hybrid Tea cultivars that are highly selected, typically tetraploid and 

highly heterozygous. Cut rose seeds do not germinate more than 50%, with hip contents 

usually ranging between one and 30 seeds. Therefore, during rose selection more 

attention should be paid to enhance the breeding efficiency itself. With the aim to 

characterize fertility markers in cut rose cultivars, a commercial breeding database (data 

1994 -2007) was analyzed. From this database a high and a low fertile pollen donor and 

a high and a low fertile seed parent were selected and crossed in partial diallel. In this 

study, vitality, germinability and morphology analyses were carried out on dried and 

fresh pollen of the four parents, that were simultaneously cultivated in Belgium and Italy. 

Results suggested a correlation between the pollen diameter and the fertility index 

obtained from the crosses. 

LP19 

L. Pipino (1,2,3,4) 

L. Leus (3) 

A. Giovannini (2) 

V. Scariot (1) 

M.C. Van Labeke(4) 


Agronomy, Forest and 

Land Management – 

University of Turin – Via 

Leonardo da Vinci 44, I- 

10095 Grugliasco (TO), 

Italy 

(2) CRA – Research Unit 


Ornamental Species – CRA 

– Corso Inglesi 508, I- 

18038, Sanremo (IM), Italy. 

(3) ILVO – Institute for 

Agricultural and Fisheries 

Research – Plant Sciences 

Unit – Caritasstraat 21, 

9090, Melle, Belgium. 

(4) Department of Plant 

Production – Ghent 

University – Coupure links 

653, 9000, Gent, Belgium 

luca.pipino@unito.it 


LP20 

Hanneke Witsenboer 

Nathalie van Orsouw 

Feyruz Yalcin 

Antoine Janssen 

Alberto Maurer 

Michiel van Eijk 

Herco van Liere 

Edwin van der Vossen 

Keygene N.V., Agro 

Business Park 90, P.O. Box 

216, 6700 AE Wageningen, 


hanneke.witsenboer@ 

keygene.com 

APPLICATION OF CROPS ® TECHNOLOGY IN A 

WIDE RANGE OF VEGETABLE AND FIELD 

CROPS 

KeyGene has developed the CRoPS technology ®1 (Complexity Reduction of 

Polymorphic Sequences) for high throughput polymorphism discovery in a wide range of 

species. CRoPS combines the power of AFLP ® 2 as a robust genome complexity 

reduction method with the power of next generation sequencing of the Roche GS FLX. 

Since CRoPS is AFLP-based, no prior sequence information is required to enable 

polymorphism discovery by sequencing genome partitions of multiple samples in 

parallel. Recently we developed an export module for the CRoPS SNP mining pipeline to 

automate the design of ready-to-order Illumina BeadExpress assays for SNP 

genotyping. 

We will present an overview of CRoPS runs performed in a number of species in 

terms of numbers of high quality putative SNPs obtained. These results show the 

efficiency of CRoPS as de novo SNP discovery technology and conversion rates of 

putative SNPs to high-throughput SNP genotyping assays. The fact that not much 

sequence information is yet available in ornamentals makes the CRoPS technology ideal 

for de novo SNP development in ornamental plants, thereby paving the way to marker 

assisted breeding. Being sequence based, the SNP genotyping platform will enable full 

integration of marker data with existing or future whole genome sequences. Sequencing 

of genomes of ornamental plants is currently within reach as sequencing costs are 

dramatically dropping with the development of new generation sequencing technologies. 

1 van Orsouw et al (2007) PLoS ONE 2 (11): e1172 

2 Vos et al (1995) Nucleic Acids Res 23: 4407-4414 

The CRoPS ® and AFLP ® technologies are covered by patents and patent applications 

owned by Keygene N.V. AFLP and CRoPS are registered trademarks of Keygene N.V. 

BeadExpress TM is a trademark of Illumina Inc. 


COOPERATIVE MARKETING – A WAY TO 

STIMULATE SALES AND CONSUMPTION OF A 

PLANT? 

While on many markets well known brands have been established, the flowers and 

plants sector developed a different approach to promote successfully the consumption of 

its products. In North and Western Europe cooperative marketing organisations have 

been set up which stimulated sales and consumption of flowers and plants. Usually, 

these activities did not focus on one product but the usage of flowers and plants in 

general, as decoration item for personal usage, as gift etc. More recently, product 

specific marketing campaigns have been set up, promoting in a cooperational approach 

the use of a certain plant or plant group. 

Why cooperative marketing – and not for a brand but flowers and plants in general – 

or even a product? Cooperative marketing organizations for flowers and plants were set 

up because of the special characteristics of plants and their market. Given the number 

and versatility of products, not one product, not even a certain product range have a 

market share which enable them to spend budgets necessary for brand marketing. 

Brand marketing is very expensive. Big brands like Coca Cola spend every year hundred 

of millions euro for advertisement. That flowers and plants are generally not seen as 

brand has a big advantage: instead of paying hundreds of million euro for expensive 

advertisement, Public Relations is the way to approach the public. Consumer magazines 

and TV shows report on how to decorate with pot plants or on the welfare of green pot 

plants. Regularly, flowers are presented as valuable gifts. Via Public relations, 

cooperative plant marketing is very cheap – and very effective. 

What presuppositions are there to conduct successfully cooperative marketing (for a 

product)? 

Presupposition for establishing cooperative marketing is a “bottle” neck. This can be 

either by a law or by a group of companies deciding to collectively conduct marketing 

measures on a voluntary base. If marketing activities are based on voluntary 

cooperations between market partners, an important presupposition is the trust among 

the participants of the group. Old conflicts should have been solved. Furthermore, there 

should be a common idea, vision or goal as well as a common decision on how to raise 

funds. 

Examples for successful marketing campaigns are e.g. the Plants of the month 

approach in the tree nursery sector, the Dutch Verrassend buiten, the European 

marketing campaign Cyclamen Colour Europe or Stars for Europe, the European 

poinsettia campaign. The goal of the marketing campaign for Cyclamen was to promote 

the outdoor usage for Cyclamen. The goal of the Poinsettia campaign is a change the 

old fashioned image and link the Poinsettia more intensively to Christmas as a time of 

extensive spendings for decoration items. 

In my presentation I will focus on how to set up cooperative marketing campaign, talk 

about budgets necessary and the effects of such campaigns. 

L29 

Susanne Lux 

Kasbach-Ohlenberg, 

Germany 

susanne.lux@meclux.de 

Session Marketing 65

L30 

Paul Roetenberg 

Platinastraat 100, Hengelo 


p.roetenberg@frederiques 

choice.com 

THE BRAND: FREDERIQUE’S CHOICE 

Mission 

Our mission is to create and grow a successful and powerful brand - a brand that stands 

for Elegance, for Luxury, for Dynamism, but also for Approachability. 

We have adopted a new way of looking at traditional marketing processes. By 

shortening the chain between producer and consumer, we will control the product and 

guarantee a consistent quality. Our objective is to bring the best quality and design from 

the source to the consumer under the brand name Frederique’s Choice. 

Let’s make flowers fashionable! 

Philosophy 

Frederique van der Wal is the personification of the Frederique’s Choice brand. 

Products marketed under the brand name invite the consumer to share in the everyday 

glamour that Frederique embodies. Frederique will personally select and guarantee the 

style and quality of every product sold under the brand name. 

A person who wants to be a discerning consumer of flowers and home products will 

only have to make one choice: FREDERIQUE’S CHOICE. 

Consumers will identify with Frederique’s approach to life, sense of style and her 

success. As one of the best-known faces in the world of lifestyle, Frederique is the ideal 

ambassador for her own brand. Her success in the world of fashion, design and beauty 

gives her the opportunity to represent her own brand: FREDERIQUE’S CHOICE, Life in 

full bloom! 

The brand is built around the following core values: 

Quality 

European style 

Selection and control from the source 

Loyalty to our brand, our people and our environment 

Support of charities 

Creating a sense of everyday luxury 

Products 

In view of the enormous potential for a new brand in the flower sector and a new 

marketing approach, Frederique’s Choice started in April 2008 by selling fresh flowers, 

bouquets and flower bulbs. There is currently no recognizable consumer brand for 

flowers. Introducing a flower brand that is identifiable and associated with quality and 

exclusive distribution is likely to have a larger impact and be of greater interest to media 

than introducing a general home and leisure brand. By starting with flowers, we 

maximize the impact of the new brand and create the right image from the start. 

Frederique’s Choice also included other flower-related products. Eventually, we will 

create our own exclusively-produced home and leisure product lines to be supported by 

the brand and will launch them in the top end of the market. 

66 Session Marketing

EARLY-FLOWERING TRANSGENIC 

CHRYSANTHEMUM PLANTS 

Chrysanthemum (Chrysanthemum morifolium) is one of the worldwide ornamental 

cultures. It is a short-day plant that blooms in autumn. When grown in greenhouse, they 

can be fooled into blooming any time of year by decreasing the amount of light they 

receive. Early-flowering cultivars are recommended to produce quality chrysanthemum 

cut flowers at lower cost of production. Flowering control should allow for shorter 

breeding cycles. 

The transition to flowering is a complex process that is regulated by many different 

mechanisms. Extensive studies in model plant Arabidopsis thaliana have revealed 

genetic and molecular mechanisms of flowering. In Arabidopsis, CONSTANS (CO) and 

FLOWERING LOCUS T (FT) act in the photoperiod pathway. They up-regulate the floral 

meristem identity genes LEAFY (LFY) and APETALA1 (AP1). These genes were 

demonstrated to be useful for shortening flowering time in certain plant species: 

constitutive expression of LFY and AP1 cause early flowering of transgenic plants. 

We had cloned three AP1-homologous genes from chrysanthemum (CDM111) and 

sunflower (HAM75, HAM92) and have become interested in the generation of 

chrysanthemum transgenic plants that over-expressed these genes. The coding region 

of each full length cDNA was fused to the CaMV 35S promoter into pGD121 binary 

vector. For Agrobacterium tumefaciens (strain CBE21) mediated chrysanthemum 

transformations, leaves from in vitro grown plants were used. Regenerating plants were 

selected on MS media containing 35 mg/l kanamycin. Transformation efficacy was about 

15%. Totally 62 independently regenerated plants carrying integrated transgenes were 

produced and used for subsequent experiments on flowering induction. 

We have demonstrated that over-expression of compositae AP1-homologs in 

transgenic chrysanthemum under long-day conditions had no effect on flowering time 

and vegetative development. In the same time, under short-day condition transgenic 

plants have started bud initiation two weeks earlier that non-transgenic control 

chrysanthemum plants. Later on, transgenic chrysanthemum plants showed colour 

earlier and resulted in earlier harvesting by 5 weeks compare to non-transgenic control 

plants. 

These results open new possibilities for genetic improvement and breeding 

chrysanthemum cultivars. 

L31 

O.A. Shulga (1) 

T. Yu Mitiouchkina (2) 

A.V. Shchennikova (1) 

K.G. Skryabin (1) 

S.V. Dolgov (2) 

(1) Centre Bioengineering 

RAS, pr. 60-let Oktyabrya 

7/1, 117312, Moscow, 

Russia 

(2) Branch of 

Shemyakin&Ovchinnikov 

Institute of Bioorganic 

Chemistry, 142290, 

Pushchino, Moscow region, 

Russia 

shulga@biengi.ac.ru 

Session Molecular breeding 67

L32 

P. Buddharak (1) 

R. Chundet (2) 

(1) School of Science and 

Technology, Naresuan 

University Phayao, Tumbol 

Maeka, Muang, Phayao, 



Faculty of Science, Maejo 

University, Chiang Mai 


su_buddha@hotmail.com 

ISOLATION AND CHARACTERIZATION OF 

FLAVONOID 3’ HYDROXYLASE (F3’H) GENE 

AND GENETIC TRANSFORMATION IN 

BUTTERFLY PEA (CLITORIA TERNATEA 

LINN.) VIA AGROBACTERIUM TUMEFACIENS 

The flavonoid 3’ hydroxylase (F3’H) gene denoted as an AsF3’H was isolated and 

characterized from petals of Ascocenda sp. The partial cDNA encoded a 264 amino acid 

polypeptide which showed a high homology to known F3’H genes, especially, F3’H from 

Gerbera hybrida. F3'H is cytochrome P450 and it is a key enzyme in the flavonoid 

pathway leading to the production of the red coloured anthocyanins. The AsF3’H 

transcript was the most abundant in petals from flowers at an early stage of development 

and levels declined as the flower matures. No transcript levels were detected in leaves 

and stem. 

In butterfly pea (Clitoria Ternatea Linn.) a genetic transformation system via 

Agrobacterium tumefaciens Was developped Therefore, the CuDFR gene was 

constructed in pBI121 and transformed into butterfly pea’s leaves, petunia (Petunia 

axillaris) and tobacco (N. tabacum) by A. tumefaciens strain AGLO.. After 4 months in 

tissue culture, the transgenic shoots were detected on selective media and the efficiency 

of transformation was 20 %, 25 % and 23 % of GUS assay respectively. Moreover PCR 

technique reveals the positive bands of GUS gene and 35S promoter. 

68 Session Molecular breeding

ORIENTAL LILY HYBRIDS ENGINEERED TO 

RESIST APHID ATTACK 

Insects such as aphids are the major animal vectors for the spread of viruses in lily 

cultivation. Viral infections in lilies lead to a decrease in bulb and flower quality and have 

a significant negative impact on the economic value of the crop. Pyrethrins and mineral 

oil are the main chemicals used to fight viral infestations mediated by aphids. The 

availability of insect-resistant cultivars would provide the means to reduce the use of 

chemicals and to allow a more sustainable cultivation. One of the modern tools in 

breeding is genetic modification. Prerequisites are a gene transfer protocol for the crop 

of interest and genes coding for the desired traits. In our laboratory an Agrobacteriummediated 

transformation protocol was developed based on Hoshi et al. (2004, Plant Cell 

Rep. 22:359). This protocol for the production of marker-free transgenic lilies proved 

successful on several L.longiflorum and on Oriental hybrid cultivars. Proteinase inhibitors 

have been used successfully in engineering insect resistance against feeding or sucking 

insects, such as aphids and thrips, after introduction of the appropriate genes. Genes 

coding for volatile repellents were also found to be effective by deterring insects. In a 

dual approach, a gene coding for a proteinase inhibitor will be combined with a gene 

coding for a monoterpenoid repellent in our marker-free binary vector, pMF2, and used 

for gene transfer to lilies. For this, a series of 22 lily cultivars, encompassing mostly 

orientals, but also longiflorums, OT and LA hybrids, have been used to induce callus on 

filaments and styles in order to test for regeneration ability and amenability to 

transformation. The first results on this will be presented. 

L33 

Frans A. Krens 

Tila R. Menzel 

Chang Liu 

Dianka C.T. Dees 

Bernadette C.E. van 

Kronenburg 




Netherlands 

frans.krens@wur.nl 


L34 

Supatida Sirisawat (1) 

Naoya Fukuda (1) 

Hiroshi Ezura (1) 

Takashi Handa (1,2) 



Science, University of 

Tsukuba, Tsukuba, Ibaraki, 

305-8572, Japan 

Tel:+8129-853-7725 

(2) School of Agriculture, 

Meiji University, Higashimita, 

Tama-ku, Kawasaki, 

214-8571, Japan 

supatidas@gmail.com 

DMMADS4, A DEF-LIKE GENE FROM 

DENDROBIUM IS REQUIRED FOR FLORAL 

ORGAN IDENTITY AND FLOWER LONGEVITY 

OF ORCHID 

Two Class B MADS-Box genes were isolated from Dendrobium moniliforme i.e. 

DMMADS4 and DMPI. DMMADS4 encodes for protein with a predicted length of 224 

amino acids; it shares 89% identity with PeMADS4 at the amino acid level. DMPI 

encodes for a 210 amino acid protein that showed 94% identity to PeMADS6 from 

Phalaenopsis. The results from RT-PCR analysis showed that DMMADS4 was 

transcribed in petals, lip and column; however, DMPI was expressed in all four floral 

whorls. Expression pattern of DMMADS4 is broader than that of PeMADS4 which is 

expressed only in the part of the lip. Therefore, DMMADS4 does not specify only lip 

development like PeMADS4. Interestingly, the expression of DMMADS4 and DMPI were 

also detected during the ovule development. A high level of expression of both genes 

was found in pre-pollinated ovules of flowers. Yeast two-hybrid analysis demonstrated 

that DMMADS4 and DMPI were able to form heterodimers. The phenotype of 

35S::DMMADS4 transformed Arabidopsis plants was indistinguishable from that of wildtype 

plants except plants size was reduced. However, in the 35S::DMPI plants, flowers 

have conversed sepals into a petaloid-like structure in whorl 1. The crossing between 

35S::DMMADS4 and 35S::DMPI resulted in the alteration of sepals into petaloid-like 

structures in whorl 1. Also, the plants maintained flower bud stage longer than wild-type 

plants. These results suggested that the B-function genes, DMMADS4 and DMPI, are 

necessary for petal development and might be necessary for flower longevity of orchid. 


OVEREXPRESSION AND SILECING OF 

KXHKN5 GENE IN KALANCHOE X 

HOUGHTONII 

K. x houghtonii (n=51), a triploid interspecific hybrid between K. daigremontiana Hamet 

& Perrier (n=17) and K. delagoensis Ecklon & Zeyher (n=34), forms epiphyllous buds on 

leaf margin notches in response to a long day photoperiod (vegetative vivipary). Several 

well known class 1 knox genes, as Kn1 from maize, STM, KNAT1 and KNAT2 from A. 

thaliana and Bkn3 from barley, play an important role in meristem formation and 

maintenance and a few reports suggest that this class of homeotic genes could be 

involved in vivipary. In attempt to characterize KNOX genes involved in vegetative 

vivipary in K. xhoughtonii, overexpression and post transcriptional gene silencing 

(PTGS) experiments of KxhKN5 (EU240661) were performed. To accomplish overexpression, 

the complete cDNA sequence of the gene (1161 bp), overdrive by 35S 

promoter and NOS terminator, was cloned in the binary vector pGreen II 

(www.pgreen.ac.uk) that contain the NPTII gene, that confer resistance to kanamycin 

(pGreen II NPTII). PTGS construct was prepared by cloning in pJM007 (Schattat et al., 

2004), a 326 bp fragment of the gene in sense and antisense orientation in the specific 

cloning sites located at the left and at the right of the PIV2 intron; silencing cassette was 

excided from pJM007 and cloned into the binary vector pGreen II NPTII; the derived 

vectors were then transferred to A. tumefaciens. Genetic transformation and selection on 

medium containing kanamycin, and cefotaxime were conducted; the regenerated shoots 

were isolated from the leaf explants and separately cultivated on propagation medium to 

establish plant clones some of which were acclimatized in greenhouse. In addition, to 

localize KxhKN5 mRNA in early fase of K. Xhoughtonii epiphylly, in situ hybridization 

was performed with digoxigenin (DIG)-labelled RNA probe according to DIG RNA 

labelling KIT ( Roche, Mannheim, Germany), with minor modifications. Overexpression 

and PTGS of KxhKN5 affect plant architecture and leaf shape as reported for class 1 

KNOX genes cloned from other species. Ectopic buds and shoots formation was not 

observed in transgenic plants overexpressing KxhKN5; furthermore the vegetative 

vivipary was reduced in the transgenic clones with extremely modified phenotype likely 

as a consequence of the leaf shape modification. 

L35 

Marina Laura 

Cristina Borghi 

Cristina Regis 

Arianna Cassetti 

Andrea Allavena 

CRA – Unità di Ricerca per 

la Floricoltura e le Specie 

Ornamentali, Corso Inglesi 

508, 18038 Sanremo (IM), 

Italy 



L36 

M. Chabannes (1) 

E. Marazuela (2) 

M. Borja (2) 

(1) CIRAD Département 

BIOS. UMR BGPI - TA 

A54/K. Campus 

International de Baillarguet. 

34398 Montpellier CEDEX 

5. FRANCE 

(2) F. PROMIVA. M-501 km 

5. Boadilla del Monte 

28660-Madrid. SPAIN 

imasd@promiva.com 

EXPRESSION OF AN ARABIDOPSIS 

ASPARTIC PROTEASE IN PELARGONIUM 

Arabidopsis thaliana transgenic plants with constitutive over-expression of the aspartic 

protease gene At2g28010 (named CDS10) showed a bushy, multi-branching dwarf 

phenotype. In order to obtain compact plants of ornamental interest with an analogous 

phenotype in Pelargonium zonale, a tall cultivar (Boda Gitana Salmon) was transformed 

to over express the A. thaliana CDS 10 gene under the 35S promoter. Twenty seven 

transgenic lines were obtained with different levels of expression after gold particle 

bombardment and regeneration. Some of them showed indeed a bushy phenotype with 

a higher number of branches and a dwarf phenotype. However, an increase in the 

number of branches correlated with a decrease in the number of petals in the flowers. So 

the plants that were of interest from the compact habit point of view, had lost the double 

flower trait, and exhibited only 5 petals/flower which were also smaller than those from 

double flowers from the non transformed plants. Intermediate phenotypes with semidouble 

flowers and higher number of branches but without a compact phenotype were 

also observed. In order to determine if it was genotype related two other cultivars were 

transformed, Mirada Violet and Mirada Simple Pink double and single flower cultivars 

respectively. Transgenic plants showed indeed a higher number of branches and single 

flowers. Even if the busy phenotype was of interest in order to get a higher number of 

cuttings/plant and a compact phenotype, the pleiotropic effects of the over-expression of 

the A. thaliana CDS 10 gene on the flowers are too strong meaning it is only of interest 

in single flowered cultivars which are a small share of the market. 


MOLECULAR MARKERS AND THEIR USE IN 

ORNAMENTALS 

In the past 20 years the use of molecular markers has gradually expanded from the field 

of scientific genetic analysis towards the implementation in commercial breeding 

programs. Therefore the last afternoon of this Eucarpia congress on “Colourful Breeding 

and Genetics” will be dedicated to the use of molecular markers in ornamental breeding. 

A general overview of the use in research and practical breeding will be given with 

examples from a variety of ornamentals such as roses, lilies and tulip. Specific attention 

will be given to the development of markers that can be used for identification of 

ornamental crops, especially if no markers are known yet for a certain species. Another 

focus will be on recent advances in sequencing technology. These are changing the way 

molecular tools are used for breeding. Enormous amounts of data can be generated in a 

very short time, and most importantly, the costs have gone down to a level that now 

makes a 1000$ genome imaginable in the near future. High density genetic maps will be 

affordable tools not only for the major crops. Fast and cost-efficient discovery of markers 

and genes for important agronomical traits is possible only when genomics data is 

combined with expert crop knowledge and accurate and reliable phenotyping. How can 

we use it in research and breeding of ornamentals? 

The company Keygene N.V. (http://www.keygene.com ) will present its view on 

marker application in ornamental research and breeding. DNA technology combined with 

the laws of inheritance is being used by Keygene to develop procedures and methods 

that can elucidate the relationship between phenotypic variation and genotypic variation, 

thereby generating knowledge of the molecular control of valuable traits. Efficient and 

effective exploitation of this knowledge is the core business of the modern plant breeder. 

Many valuable traits have a complex inheritance in plant populations and therefore the 

molecular control of this type of traits has a complex nature as well. This presents 

serious challenges for the future and requires integration of different knowledge levels as 

well as breeding strategies and breeding schemes. 

The afternoon will be closed by a round table discussion. During the congress it will 

be possible to submit specific questions to the organizers (a questionnaire will be 

provided in the congress bag). The questions will be discussed by participants and 

organizers. 

L37 

Paul Arens (1) 


Sjaak van Heusden (1) 

Hanneke Witsenboer (3) 

(1) Wageningen UR Plant 

Breeding P.O. Box 16, 

6700 AA Wageningen, the 

Netherlands 

(2) ILVO-Institute for 

Agricultural and Fisheries 

Research, Plant Science 

Unit, Caritasstraat 21, 9090 

Melle, Belgium 

(3) Keygene N.V., Agro 

Business Park 90, P.O. Box 

216, 6700 AE Wageningen, 


Paul.arens@wur.nl 

Sjaak.vanheusden@wur.nl 

Hanneke.witsenboer@ 

keygene.com 

Jan.deriek@ilvo. 

vlaanderen.be 

Workshop Molecular markers and their use in ornamentals 73

7. Abstracts of posters 

P1 

W. Amaki 

S. Takeuchi 

Graduate School of 

Agriculture, Tokyo 

University of Agriculture 

1737 Funako, Atsugi, 

Kanagawa 243-0034, 

Japan 

amaki@nodai.ac.jp 

ISOLATION OF FEMALE GAMETE CELLS 

FROM UNPOLLINATED OVULES IN LILIUM 

Ovules obtained from opened flowers of Oriental hybrid ‘Casa Blanca’, LA hybrid ‘White 

Tycoon’, Lilium × formolongi ‘Hakuryu’ and L. formosanum were wounded by a surgical 

blade, and then treated with an enzyme mixture. Afterwards, ovule sections were 

dissected using a handmade glass needle and the tip of a 30G short needle syringe 

under a stereomicroscope to remove the outer and/or inner integument. The isolated 

embryo sac with nucellar tissue was cut off by the 30G short needle syringe, and was 

then pushed off by a macromanipulating system to isolate female gamete cells. The 

isolated gamete cells could be easily captured and handled by a microcapillary system 

that was connected to an electric micropump. The combination of a surgical cutting with 

the use of an enzymatic treatment, shortened significantly the total operating time for the 

isolation of female gamete. However, the optimal incubation time for the enzymatic 

treatment differed between cultivars and species. It varied from 1 to 1.25 and 2.5 hrs for 

L. × formolongi ‘Hakuryu’, L. formosanum and L. ‘Casa Blanca’, respectively. No isolated 

gamete cells could be obtained from ovules of ‘White Tycoon’ due to the fact that the cut 

flowers were stored at low temperature for several days. 

74 Session Posters

HERITABILITY OF COLD TOLERANCE IN 

GLADIOLUS 

Gladiolus xhybridus are not cold tolerant (winter hardy) in northern temperate climates, 

such as USDA Winter Hardiness Zones 3-4, with corms capable of surviving -10°C to - 

12°C temperatures. The University Of Minnesota Gladiolus Breeding Program has an 

objective to screen germplasm (wild species, intra- and inter-specific hybrids) for use as 

parents to determine the extent of cold tolerance in the genus and its heritability for 

future release of winter hardy cultivars that do not require fall digging and overwintering 

in storage (>0°C). Wild species and hybrids were tested to quantify LT 50 (the lethal 

temperature at which 50% of the progeny were killed) using controlled freezing tests with 

acclimated corms. Several hardy parents were found (LT 50

P3 

Alice Noemí Aranda- 

Peres 

Carolina Cassano Monte 

Bello 

Adriana Pinheiro 

Martinelli 

Universidade de São Paulo, 

Centro de Energia Nuclear 

na Agricultura, Av. 

Centenârio, 303, Piracicaba 

– São Paulo, Brazil 

adriana@cena.usp.br 

INTERSPECIFIC HYBRIDIZATION OF 

BRAZILIAN VRIESEA SPECIES 

Bromeliaceae are endemic to Brazil, where forty percent of the species and seventy 

percent of the genus of this family are present. However, this important diversity has 

been threatened by extractivism. To generate new ornamental material, which can help 

to avoid excessive extractivism, hybridizations between different genetic accesses are 

important. In the present work, six species of bromeliads from the Brazilian Atlantic 

Forest and two commercial hybrids were hybridized. Pollen-receptor flowers were 

emasculated before anther dehiscence, following hand pollination with pollen from 

pollen-donor plants. Controlled crosses were done using plants from the same species 

as well as interspecific crosses and crosses between species and commercial hybrids. 

Capsule formation was observed after 60 days. Crosses between V. carinata x V. 

paraibica showed 100% capsule formation, independently of the pollen donor/receptor 

species. Reciprocal crosses using V. friburguensis, V. incurvata and V. simplex also 

presented high percentage of capsule formation. Seeds were collected when capsules 

matured and were placed in ¼ MS culture medium (Murashige & Skoog, 1962) for in 

vitro germination. Seedlings were grown in vitro and RAPD analyses were performed to 

confirm the hybrid character of the seedlings. The results indicate the efficiency of 

crosses with Vriesea species and their potential use for the production of new options for 

the ornamental bromeliad market. 

76 Session Posters

KARYOTYPE ANALYSIS OF THREE IRANIAN 

NATIVE IRIS SPECIES 

There are several native species of Iris in Iran that has not been studied cytogenetically 

yet. Therefore, more cytological and molecular research is needed for their classification 

and identification. In this study, karyotypic analysis was carried out in three native Iranian 

Iris species: I. caspica, I. spuria and I. meda. I. caspica consisted of 22 pairs of 

chromosomes: 4 pairs of m-type, 8 pairs of sm-type and 10 pairs of acr-type. The pair 

chromosome 5 of this species had satellites. Total form percentage (TF%) in this species 

was 30.93%. In studied metaphase cell in this specie arm ratio of the chromosomes 

ranged between 1.10±0.04 and 5.30±0.88 and all of the chromosomes with arm ratio 

lower than 1.7 were metacentric. The mitotic metaphase cells of I. spuria consisted of 22 

pairs of chromosomes; including 13 pairs of m-type and 9 pairs of sm-types without 

satellites. The TF percentage in this specie was 39.18%. In studied cells in this specie 

arm ratio of the chromosomes ranged between 1.18±0.03 and 2.94±0.7. The somatic 

chromosome complement of the mitotic metaphase cells of I. meda consisted of 10 pairs 

of chromosomes; including 1 pair of sm-type, 7 pairs of acr-type and 2 pairs of t-type 

without satellites. The TF percentage in this specie was 16.85% and arm ratio of the 

chromosomes ranged between 5.08±0.22 and 7.23±0.04. Regarding to karyotype 

formula of three iris species the polyploidy and chromosomal structure rearrangement 

have an important role in iris speciation procedure. Also the existence of many different 

chromosomes in individual species may imply the adaptation of this species with 

ecological circumstances. This study showed that I. meda species that was collected 

from mountainous zones had bigger chromosome as well as bigger genome than other 

species was collected from warmer climate. 

P4 

Mostafa Arab (1) 

Vahid Rahimi (1) 

Shirin Dianati (1) 

Issa Zarif (2) 


Horticulture, Abouraihan 

Campus, University of 

Tehran, Tehran, Iran 

(2) Genetics and Genetic 

Resources Research 

Department, Seed and 

Plant Improvement 

Institute, Karaj, Iran 

mosarab@ut.ac.ir 

Session Posters 77

P5 

M. Ardelean (1) 

Doina-Angela Pui (2) 

Mirela Cordea (1) 

(1) University of Agricultural 

Sciences and Veterinary 

Medicine, Cluj-Napoca, 

3-5 Manastur Street, 

Romania 

(2) “Babeş-Bolyai” 

University, Mihail 

Kogălniceanu Street, Cluj- 

Napoca, Romania 

mardelean@usamvcluj.ro 

EXPERIMENTAL RESULTS ON GENETICS 

AND BREEDING OF GILLYFOWERS 

(MATTHIOLA GENUS) 

Two species of Matthiola (M. incana L. and M. longipetala var. bicornis) as well as eight 

cultivars of M. incana were tested in three successive years (2003-2005) in Cluj-Napoca, 

Romania, concerning the homogeneity of their distinctive plant characters (plant height, 

no of inflorescences/plant, no. of flowers/inflorescence, flower diameter, start of 

blooming and persistence of flowering stage) which are considered of interest in 

breeding programs for ornamental purposes in this genus. On the basis of the obtained 

results, four cultivars of gillyflower (M.incana L.) were chosen as valuable genitors for the 

above mentioned characters and employed in an incomplete diallel crossing system in 

2005, F 1 hybrids as well as their parental cultivars being grown in Cluj-Napoca in 2007 

and 2007. Heritability both in wide (H) and narrow (h 2 ) sense was computed for the 

same characters as those analyzed in parents and phenotypic and genotypic 

correlations among all possible pairs of characters were evaluated and discussed in view 

of their efficiency on indirect/tandem selection. Additive and nonadditive effects of the 

poplygenes involved in the inheritance of the six analyzed characters were revealed by 

computing GCA and SCA values of the genitors involved. 

The values of H = 0.55-0.84 were rather high suggesting a medium to very good 

phenotypic expression of the analyzed characters in F 1 hybrids of gillyflower. On the 

other hand, the medium and low values of the narrow sense (h 2 ) heritability for most the 

analyzed characters (except for start of blooming) revealed the fact that, in the case of 

these quantitative characters, most often high heritability in wide sense was 

accompanied by low or, at most, medium heritability in narrow sense. Quite a few of the 

analyzed pairs of characters were found significantly correlated at the phenotypic level, 

in the F 1 populations of gillyflower but out of these only in four pairs (plant 

height/diameter of flower, diameter of flower/no. of flowers in inflorescence, plant 

height/persistence of flowering stage, start of blooming/persistence of flowering stage) 

significant correlations were noted at the genotypic level as well. For all the analyzed 

characters, significant values of GCA effects were found this being true also for values of 

SCA effects, except for no. of flower/inflorescence in which the SCA effects were not 

significant. 

78 Session Posters

THIN CELL LAYER SOMATIC HYBRIDIZATION 

BETWEEN CALADIUM HUMBOLDTII SCHOTT 

‘PHRAYA SAVET’ AND C. BICOLOR (AIT.) 

VENT. ‘SUVARNABHUM’ 

Characters of new clones from somatic hybridization between Caladium humboldtii cv. 

‘Phraya Savet’) and C. bicolor cv. ‘Suvarnabhum’ using thin cell layer technique from in 

vitro culture were observed. Callus were induced from unexpanded leaf segments 

cultured on modified Murashige and Skoog medium (MS) supplemented with 2.69 µM 1- 

Naphthalene acetic acid (NAA) and 17.76 µM N 6 -Benzyladenine (BA) for 4 – 5 months 

with subculturing every 6 weeks. Each thin cell layer of callus, about 1 mm thick, from 

both caladiums was alternately laid on the top of each other for 8 layers. Subsequently, 

the combination of thin cell layers was cultured on the same medium. The regenerated 

plantlets were grown in glasshouse conditions. From 3 morphological groups (leaf 

pattern, leaf colour and petiole), the regenerated caladium plants were found dissimilarly 

to both original caladiums at 85 percent with 8 types of different characters. Somatic 

hybridization between C. humboldtii cv. ‘Phraya Savet’ and C. bicolor cv. ‘Suvarnabhum’ 

gave rise to a number of most hybrids with all conserving C. bicolor characters. 

However, variations of each caladium from in vitro culture were compared to 

regenerated hybrids. 

P6 

M. Areekijseree 

C. Thepsithar 

A. Thongpukdee 

U. Somkanae 

Department of Biology, 



Sanamchandra Palace 

Campus, Muang, Nakhon 

Pathom 73000, Thailand 

maijackee@yahoo.com 

Session Posters 79

P7 


Danny Esselink (1) 

Yolanda Noordijk (1) 

Linda Kodde (1) 

Lysbeth Hof (2) 

Willem Wietsma (2) 

Ben Vosman (1) 

(1) Plant Breeding, 

Wageningen UR, P.O. Box 

16, 6700 AA Wageningen, 


(2) Unit Varieties & Trials, 

Naktuinbouw, P.O. Box 40, 

2370 AA Roelofarendsveen 


paul.arens@wur.nl 

IDENTIFICATION OF CARNATION VARIETIES 

USING MICROSATELLITE MARKERS 

As in many ornamentals, also in carnation the number of varieties in common knowledge 

is large. Variety registration and protection in carnation is based on morphological 

characteristics, described in the UPOV Test Guidelines (document TG/25/8). 

Identification throughout the chain from breeder to consumer depends on the availability 

of methods that can use plant material from different stages and organs. Furthermore, in 

situations where a suspicion of an infringement of a PBR has been raised, there may be 

a need for a quick and reliable comparison of varieties before a lengthy trial for 

comparison of DUS characteristics is considered. Therefore, complementary 

approaches, such as the use of microsatellite markers, are being evaluated and 

molecular databases are being constructed. Microsatellites are highly polymorphic and 

have the advantage of providing co-dominant markers based on PCR technology. In a 

sequenced tagged microsatellite site (STMS) approach, they produce simple banding 

patterns, especially suitable for automated and objective description of plant varieties 

which are easy to store in a database. New varieties or new markers can easily be 

added to an existing database. 

In this paper, we present the results of our study on the use of microsatellite markers 

from Dianthus caryophyllus L. for the characterization of carnation varieties as well as 

the construction and evaluation of a molecular database. 

80 Session Posters

INTRASPECIFIC AND INTERSPECIFIC 

HYBRIDIZATION OF TROPICAL LILY (LILIUM 

SPP.) 

Lily (Lilium sp) is one of important cut flowers in Indonesia. Demands are increasing over 

the years. Lily has been utilized for flower arrangement, bouquet, and decoration in 

important events such as wedding ceremonies and International Conferences which are 

very often held in Bali. Production of Lily in Bali in particular, or in Indonesia in general 

are still limited and market are still dominated by imported Lily. here is an urgent need to 

produce superior varieties of Lily locally. Lily grown in Indonesia are mainly local 

cultivars which has limited variation in term of colour, shape and shelf life. However, 

local Lily has the benefit of well adaptation in tropical climate and produces more flowers 

in one stem compare to introduce cultivars. Therefore, research need to be done to 

increase Lily variation through breeding, so that Lily varieties with superior and 

preferable characteristics, such as increase flowering period, flower shape and size, 

more colour and resistance to disease can be obtained. This research project attempted 

to do intraspecific and interspesific hybridization of local lily. The short term objective of 

this project is to obtain viable crosses via of embryo rescue technique, while long term 

objective is to obtain new cultivars with unique and superior characteristics which are 

favored by consumers. This research is currently underway in Bali. Materials for crosses 

were obtained from collection of ‘Eka Karya’ Botanical Garden in Bali and flower 

growers. For intraspecific crosses, 3 cultivars of local lilies (orange, yellow and brown) 

were crosses reciprocally. For interspecific crosses, two cultivars of local lily (yellow and 

light brown) were employed as mother plants, while pollen from imported lilies were used 

as male plants. To avoid embryo abortion in interspecific crosses, embryo will be 

harvested before the seed mature and then transfer to tissue culture media. Crosses in 

currently underway. It is expected that viable plantlet could be obtained in the next year 

for characteristics evaluation. 

P8 

Ida Ayu Astarini 

Yunita Hardini 

Biology Department, 

Faculty of Mathematics and 

Natural Sciences, Udayana 

University, Bali, Indonesia, 

80364 

idaastarini@yahoo.com 

Session Posters 81

P9 

Swati Barche (1) 

Kamal S. Kirad (2) 

D.B. Singh (3) 

(1) JNKVV, Jabalpur , 

College of Agriculture, 

Tikamgarh, M.P. India 

(2) Department of Welfare 

& Agril. Devlopment, 

Tikamgarh, MP, India 

(3) Allahabad Agricultural 

Institute-Deemed 

University, Allahabad, U.P 

sbkdap07@rediffmail.com 

GENETIC VARIABILITY IN GLADIOLUS 

Gladiolus or sword lily is commercially significant bulbous ornamental crop which is 

grown either as cut flower or garden display. Improvement in gladiolus required genetic 

variability, its heritability and genetic progress. Also, knowledge of association between 

yield and its component traits themselves can improve the efficiency of selection. The 

present study was undertaken to observe the phenotypic variability into its heritable and 

non-heritable components with suitable genetic parameters viz., phenotypic and 

genotypic coefficient of variation, heritability, genetic advance. Fifteen elite genotypes of 

gladiolus were collected from various places and conducted experiment at research 

farm, Department of Horticulture, Allahabad Agricultural Institute-Deemed University, 

Allahabad during 2005 in Randomized Block Design with three replications. High 

genotypic coefficients of variation (GCV) and phenotypic coefficient of variation (PCV) 

estimates were found for number of cormels per corm. High heritability with high genetic 

advance was observed for number of cormels per corm, days for spike emergence and 

spike weight. Number of florets per spike was significantly and positively correlated with 

number of shoots per corm and duration of flowering. Spike length and diameter of corm 

was significantly and positively correlated with spike weight but negatively correlated 

with duration of flowering. 

82 Session Posters

PHYLOGENETIC ANALYSIS OF ARBUTUS SPP 

BY MORPHOLOGICAL CHARACTERISTICS 

AND MOLECULAR MARKERS 

Morphological characteristics and the method of Random Amplified Polymorphic DNA 

Polymerase Chain Reaction (RAPD-PCR) was used to study the diversity of Arbutus 

andrachne and A. unedo individuals from two different regions of collection, Kalamos 

and Varympompi, prefecture of Attici (Greece), and individuals of an Arbutus sp. with 

intermediate morphological characteristics found in Kalamos. The morphological 

characteristics that were studied were: bark morphology, leaf shape, flowers and season 

of flowering, fruits and season of fruiting. The bark of A. andrachne plants and of those 

with intermediate characteristics was smooth and cinnamon-red in colour, peeling in long 

paper strips revealing a grey-green internal. Oblong areas of dark red colour were found 

on the bark of plants with intermediate morphological characteristics. A. unedo bark was 

rough and shreddy dull brown or ash-grey and occasionally peeling in small flakes 

revealing chestnut-coloured internal. The leaves of A. andrachne were ovate to oblong, 

entire, occasionally pointed, and of A. unedo elliptic to oblanceolate, toothed, pointed, 

while of the intermediate plants were elliptic to obovate, occasionally pointed and often 

toothed. The plants with intermediate characteristics fructified seldom contrary to the rich 

fructification of A. andrachne and A. unedo. Four 10-mer oligonucleotide arbitrary 

primers were used to amplify genomic DNA and 36 reproducible polymorphic fragments 

were generated. The degree of genetic similarity was calculated and the dendrogram of 

seven individuals was established. A genetic variation among individuals that bring 

intermediate morphological characteristics and those of A. unedo and A. andrachne was 

indicated, confirming the morphological variations observed. This allows the statement 

that it is another species, at least for the primers that were used, possibly the one 

reported in the bibliography as a natural hybrid between A. unedo and A. andrachne, 

named Arbutus x andrachnoides. 

P10 

Konstantinos F. 

Bertsouklis (1) 

Maria Papafotiou (1) 

Katerina Biniari (2) 

(1) Laboratory of 

Floriculture and Landscape 

Architecture, Department of 

Crop Science, 

(2) Laboratory of Viticulture, 

Department of Crop 

Science, Agricultural 

University of Athens, Iera 

Odos 75, 118 55 Athens, 

Greece 

mpapaf@aua.gr 

Session Posters 83

P11 

Paul J.J. Bijman 

Jan F. Demmink 

Jaap M. van Tuyl 

Wageningen Box 16, 6700 

AA Wageningen, the 

Netherlands 

Paul.bijman@wur.nl 

BREEDING FOR NEW CUTFLOWER AND 

POTPLANT VARIETIES IN ZANTEDESCHIA 

(CALLA) 

Zantedeschia, also named Calla, belongs to the family of Araceae and is originally from 

South Africa. In this breeding programme we only use the tuber types of Zantedeschia 

section Aestivae and no root stock types of Zantedeschia aethiopica or odorata (Singh et 

al., 1996). The crop is increasing in popularity. The flowers of Zantedeschia distinguish 

themselves for flower form, colour, long vase life and can be transported under dry 

conditions. Tubers can be stored during a long period of time and therefore it is possible 

to produce flowers over a long period. Zantedeschia as a potplant shows a great 

promise due to the wonderful flower colours. 

The poster will give a description of the pollination technique we use, technique for 

testing pollen germination and criteria we use for selecting crossing parents to make new 

varieties in cut flower and potplant types. 

Singh, Y., H. Baijnath, et al., (1996). Taxonomic notes on the genus Zantedeschia 

Spreng. In Southern Africa., South African Journal of Botany 62(6): 321-324. 

84 Session Posters

INDUCTION OF COLOURED CALLUS FROM 

LAWSONIA INERMIS SYN. ALBA 

Tissue culture of Lawsonia inermis, syn. alba (henna) were carried out to induce 

coloured callus formation as well as the plant regeneration in vitro. Various explants from 

aseptic seedling were used such as leaf, stem, and root. These explant were cultured on 

Murashige and Skoog (MS) medium with different combinations and concentrations of 

hormones. The hormones used were 2,4-dichlorophenoxyacetic Acid (2,4-D), 

Napthalene Acetic Acid (NAA), Benzyl Aminopurine (BAP), Kinetin, and Indole acetic 

Acid (IAA). This studies focused on getting coloured callus from the explants which 

contain the important secondary metabolites such as lawsone and isoplumbagin. These 

secondary metabolite are useful for medicinal purposes. It can affect the body by slowing 

down the heart rate, reducing blood pressure,fever and pain, and also by acting as a 

sedative. Beside that, the economic importance of this herbal species is also used in 

cosmetic products such as hair conditioner, anti-dandruff, nail strengthening effects, and 

also as a sunscreen 

P12 

Fatimah Binti 

Abdul Rahiman 

Rosna Mat Taha 

Institute of Biological 

Sciences 

Faculty of Sciences, 

University of Malaya, 50603 

Kuala Lumpur, Malaysia 

fatimah_rahiman@ 

hotmail.com 

Keywords: Lawsonia inermis, in vitro, medicinal plant, Murashige and skoog (MS) 

ornamental plant, 2,4-dichlorophenoxyacetic Acid (2,4-D), Napthalene Acetic Acid 

(NAA), Benzyl Aminopurine (BAP), Kinetin, Indole acetic Acid (IAA) and coloured callus. 

Session Posters 85

P13 

Thomas Borchert 

Katja Krueger 

Annette Hohe 

Leibniz-Institute of 

Vegetables and 

Ornamental Crops (IGZ), 

Kuehnhaeuser Str. 101 

99189 Erfurt, Germany 

borchert@erfurt.igzev.de 

FLOWER TYPES IN THE ORNAMENTAL CROP 

CALLUNA VULGARIS – MORPHOLOGICAL 

AND MOLECULAR INVESTIGATIONS 

Calluna vulgaris is one of ~ 11,500 species in the Ericales family which display an 

enormous diversity of flower types (Schönenberger et al. 2005). C. vulgaris itself 

features several different flower types, one of which is the so-called ‘bud-flowering’ 

phenotype. 

‘Wild-type’ flowers comprise four whorls of flower organs: one outer whorl of four 

coloured leaves, an inner whorl of four coloured leaves that are slightly grown together, 

eight stamens and four carpels. In contrast, ‘bud-flowering’ phenotypes display two 

identical whorls of four coloured leaves each that are not grown together. Moreover, the 

stamens are missing, which we assume to be at least one reason for the developmental 

arrest in the bud stage. Since an investigation regarding the flower organ identity of C. 

vulgaris (McClintock 1986) remained vague to the authors, we investigated the flower 

morphology in depth and started first analyses on the molecular basis of flower organ 

identity in C. vulgaris. 

Histological analyses demonstrated the time-course of organ development in young 

flower buds and revealed that in ‘bud flowering’ phenotypes stamens are not only 

degenerated but totally missing. Scanning electron microscopy (SEM) was applied for 

analyses of the cell surface structure of the coloured leaf whorls. Here, we were able to 

identify differences of the cell structure between the two coloured leaf whorls of ‘wildtype’ 

flowers which is a strong hint to identify the outer whorl as sepals and the second 

one as petals. Analogous analyses of these organs in ‘bud-flowering’ phenotypes 

supports the hypothesis, that here both whorls of coloured leaves are to be classified as 

sepals. Thus, in this flower phenotype, not only a loss of stamens has occurred but also 

a change of organ identity in the second whorl. 

Therefore, we are aiming at isolating MADS-box genes in C. vulgaris that might 

control these changes in flower architecture. 3’-RACE-PCR was applied in order to 

amplify MADS-box homologues from ‘wild-type’ mRNA in C. vulgaris. Until now, cloning 

and sequencing of RACE fragments led to the identification of B-gene homologues 

(DEF, GLO). 

McClintock (1986) Acta Hort. 182:277-283 

Schönenberger et al. (2005) Int. J. Plant Sci. 166(2):265-288 

86 Session Posters

IN VITRO PLANT REGENERATION AS A TOOL 

TO IMPROVE ORNAMENTAL CHARACTERS IN 

PASSIFLORA SPECIES 

The genus Passiflora, comprising about 500 species of vines, lianas and small trees, is 

the largest in the Passion flower family (Passifloraceae). Several species are grown in 

the tropics for their edible fruits (Passiflora edulis Sims.) and many others are grown 

either outdoors, in the warmer parts of the world, or in the glasshouses, for their exotic 

flowers. Passiflora exhibits several unique floral features, including multiple series of 

brightly coloured coronal filaments and elaborate floral nectary structures, which are of 

particular interest for the floricultural market. With the aim to exploit the ornamental value 

of some Passiflora species, a collection was settled at the CRA-FSO, in Sanremo 

(indoors). Nodal segments, floral buds and auxillary tendrils from greenhouse-grown 

plants were sterilized and in vitro propagated. Direct shoot regeneration was achieved 

from P. ‘Guglielmo Betto’, P. x allardii Lynch (P. quadrangularis x P. caerulea 

“Constance Eliott”) and from P. trifasciata Lemaire tendrils cultivated on MS medium 

containing, either 4.4 M 6-benzylaminopurine (BAP) and 11.42 M indoleacetic acid 

(IAA). P. foetida L. cv. Hastata shoots were regenerated from immature flower 

organogenetic callus, developed on MS medium supplemented with 4.4 M (BAP) and 

11.42 M (IAA). The in vitro regenerated plants were successfully acclimatized in the 

greenhouse. At flowering only P. foetida L. cv. Hastata regenerated plants showed 

morphological alterations in flower and fruit external bracts. Citological and molecular 

analyses of in-vitro cell clones and regenerated plants will be performed in order to test 

the existing genotypic variability. Immature flower shoot regeneration could be used in 

ornamental Passiflora for exploiting somaclonal variability in genetic improvement 

studies. 

P14 

Luca Braglia 

Laura De Benedetti 

Luca Pipino 

Annalisa Giovannini 

Antonio Mercuri 

CRA-FSO Research Unit 


Ornamental Species 

Corso Degli Inglesi 508, 

18038 Sanremo – ITALY 


Session Posters 87

P15 

A.C.R. Castro (1) 

E.B. Morais (1) 

I.C.S. Mourão (1) 

A.C.P.P. de Carvalho (1) 

V. Loges (2) 

(1) Embrapa Tropical 

Agroindustry Corporation, 

R. Dra. Sara Mesquita 

2270, 60511-100 Fortaleza 

(CE) Brazil 

(2) Federal Rural of 

Pernambuco University, Av. 

D. Manoel de Medeiros s/n, 

52171-900 Recife (PE) 

Brazil 

cecilia@cnpat.embrapa.br 

ORNAMENTAL FOLIAGE POTENCIAL OF 

THIRTY FOUR ANTHURIUM ACCESSIONS 

The Anthurium genus comprises more than 600 species, most of them with ornamental 

potential. However, only Anthurium andreanum is remarkable in the floriculture industry, 

with an enormous commercial importance as cut flower. In the last few years, the 

commercialization of new foliage has been growing up. It is necessary to introduce new 

foliage crops in culture to reach market space and Anthurium species are an excellent 

option for cut foliage exploration, nevertheless, in Brazil, this is an incipient activity. The 

Brazil Northeast has great potential for these species cultivation, for local consumption 

or exportation, due to ideal climatic condition, excellent geographical localization. The 

aim of this work was evaluate the ornamental potential for cut foliage of 34 anthurium 

genotypes, from different Brazilian ecological regions, through morphological descriptors 

and phenological aspects. Anthurium collection has been carried out at Embrapa 

Tropical Agroindustry Corporation, located in Fortaleza-CE, Brazil. Plantlets from 

promising cut foliage accessions were obtained from sexual and vegetative propagation. 

The accessions were cultivated in plastic pots, containing commercial substrate under 

80% of artificial shade. The accessions differ widely in leaf form, inflorescence size and 

stem length. It was also observed variation in: imature spadix colour (varing from light 

green to purple) and spadix diameter (ranging from 2,5 to 15,0 mm). Spathe length and 

width ranged from 3,0 to 25 cm and 1,5 to 5,cm respectly. The main nervure and pulvine 

were proeminent in all accessions. Peciolus colour was green in almost accessions, 

sometimes reddish, peciolus length ranging from 3,0 to 24,0 cm. Erect infructescences 

containing oblong to obvoid, white to purple berries were observed. About phenological 

aspects: accessions took 20 to 60 days to form complete developed leaves and 40 to 80 

days for inflorescence emittion and complete development, and the total cycle from 

inflorescence emission to fructification took more than 100 days in all accessions. The 

peciolus length and inflorescence position in relation to the leaves are important 

atributtes to use indication. All accessions presented leaves with more than 40 days of 

shelf life. The descriptions obtained could indicate ten accessions with excellent foliage 

characteristics for commercial exploration and material for future breeding programs. 

88 Session Posters

ANTHURIUM CONSERVATION STRATEGY 

USING 6-BENZILAMINOPURINA (BAP) FOR 

MULTIPLICATION 

P16 

Anthurium Schott. (Araceae) understand about 1000 species, usually herbs, epiphytes, 

natives of Tropical America. Most of them are ornamental, for its beautiful, exotic and 

long-lasting inflorescence and foliage. Just few species of anthurium are in cultivation, 

and many of them are vulnerable to antropic action. Obtained directly from the nature, 

sometimes just few plants are disponible or it is hard and slow to growth. A specific 

production system of plantlets, is a essential stage to the introduction of these species to 

evaluation, characterization and cultivation experiments. Anthurium could be sexually 

propagated, by seeds, resulting in heterogeneous populations and asexually, by stem 

section or shoot separation, but it’s a slow process. Therefore, in vitro technology has 

considerable potential to plant proliferation, plantlets production in large scale, free of 

pathogens. The aim of this work was evaluated in vitro proliferation rate of Anthurium 

plowmanii and Anthurium longipes, in modified Pierik medium, with different 

concentration of BAP (0 - control; 2,22; 4,44; 6,66 e 8,88 μM). The stem segments 

explants, obtained from well established plants, were inoculated in flasks with 30 mL of 

culture medium, and maintained in growth room at 25 ± 1ºC temperature, at 30 μmol.m - 

2 s -1 light intensity and 16 hours of photoperiod. It was used a completely randomized 

A.C.R. Castro 

I.C.S. Mourão 

F.A. S. Aragão 

A.C.P.P. Carvalho 

Embrapa Tropical 

Agroindustry Corporation, 

R. Dra. Sara Mesquita 

2270, 60511-100 Fortaleza 

(CE) Brazil 


design, in 5 x 10 factorial arranging, containing 2 replicates in a flask. After 28 days of 

inoculation, leaves and shoots numbers and proliferation. 

Session Posters 89

P17 

Fangyun Cheng (1) 

Dong Zhang (1) 

Shiliang Zhou (2) 

(1) College of Landscape 

Architecture, Beijing 

Forestry University, Beijing 

100083, China 

(2) Laboratory of 

Systematic and 

Evolutionary Botany, the 

Chinese Academy of 

Science, Beijing 100093, 

China 

chengfy8@263.net 

CROSSES FOR THE ADVANCED 

GENERATION OF TREE PEONIES AND 

HYBRID IDENTIFICATION BY AFLP MARKERS 

Reciprocal crosses between Paeonia rockii hybrids (RH), P. ostii ‘Feng Dan Bai’ (FDB) 

and P. ×lemoine ‘High Noon’ (HN) were made in 2006 and 2007 for breeding advanced 

generation tree peonies. The results from 25cross-combinations and 434 pollinated 

flowers showed that both RH×HN and FDB×HN were fertile with respectively 12.17and 

4.45 seeds harvested in each flower averagely, but their reversed crosses were almost 

infertile completely, indicating that HN could should be very promising as pollen supplier 

in cross-breeding for the advanced generation of tree peonies. And 22 samples selected 

at random from the seedlings of 7 cross combinations of RH×HN were identified by 

AFLP makers to confirm the significance of this technique in early identification of the 

hybrids. Totally 1141 bands including 1051 polymorphic ones (92.7%) were amplified by 

9 pair primers. The data of AFLP markers showed that the F 1 seedlings possessed some 

bands detected in both of the parents together, some specific to the maternal or paternal 

parents, but fewer bands didn’t exist completely in the parents. Combined with UPGMA 

analysis, these results suggested that the seedlings should be true hybrids, most of 

which were more relative to the maternal parent than to the paternal one with the novel 

variation happened in a few cases, and AFLP makers were effective in early 

identification of tree peony hybrids. Therefore, based on a higher seed setting as cross 

compatibility and the most of F1 seedlings were identified as true hybrids by AFLP 

makers, the cross RH×HN were recommended as a promising combination for the 

breeding advanced generation. 

90 Session Posters

AGROBACTERIUM-MEDIATED TRANS- 

FORMATION OF DENDROBIUM ORCHID 

A transformation protocol for dendrobium orchid was established by using immature 

protocorms as a target material for Agrobacterium inoculation. Seeds were obtained 

from the cross between two elite clones of Dendrobium nobile, ‘Cinderella’×‘True love’, 

and germinated on New Dogashima medium containing 10 g/l sucrose without any plant 

growth regulators. Three-weeks-old protocorms were subjected to co-cultivation with 

Agrobacterium tumefaciens EHA101 containing plasmid pIG121-Hm that harbored 

genes for -glucuronidase (gus), hygromycin phosphotransferase (hpt) and neomycin 

phosphotransferase II (nptII). More than 40% of the inoculated protocorms were 

recovered 2 months after selection of transformed protocorms on medium containing 20 

mg/l hygromycin. Transformation efficiency was increased by prolonged period of 

inoculation (~3 h) with Agrobacterium suspension culture. Integration of transgenes was 

confirmed by PCR analysis and Southern hybridization. Stable expression of gus gene 

was indicated by histochemical GUS assay in the leaves and roots of transgenic plants. 

P18 

D.P. Chin 

M. Mii 


Technology, Graduate 

School of Horticulture, 

Chiba University, 648 

Matsudo, Matsudo, Chiba 

271-8510, Japan 

dpchin@faculty.chiba-u.jp 

Session Posters 91

P19 

Jong-Cheol Choi (1) 


Seung-Tae Kim (2) 

Gwang-Yeon Gi (3) 

Tae-Ho Han (1,4) 

(1) Dept. of Plant 

Biotechnology, Chonnam 

National University, 

Gwangju 500-757, Korea 

(2) National Horticultural 

Research Institute, RDA, 

Suwon 440-760, Korea 

(3) Jeollanamdo 

Agricultural Research and 

Extension Servies, Naju 

520-830, Korea 

(4) Institution of Agricultural 

Science and Technology, 

Chonnam National 

University, Gwangju 500- 

757, Korea 


STUDY ON INCREASING ROSE SEED 

GERMINATION 

Rose is one of the most important cut flowers all over the world as well as Korea. 

Growers in Korea are exclusively relying on foreign cultivars. Rose breeding program 

started a decade ago, and lack of experience brought in series of topics that required 

attention from Korean researchers. One of the topics is the low germination rate in rose 

around 20%. High amount of substances that inhibit germination were found in the 

pericarp of the achene. 

Four different treatments such as grinding (0, 5, 10min), UV-irradiation (0, 5, 10, 20, 

30min), immersion in sulphuric acid (0, 5, 10min), and in microorganism (klebsiella 

oxytoca treatment for 0, 1, 48h), were applied to improve germination rate. 

Only microorganism treatments (1 and 48h) were successful on improving 

germination rate by two times compared to the control. At the moment, we are testing 

microorganism in various ways together with temperature alterations. 

92 Session Posters

REGULATION AND QUALITY OF FLOWERING 

IN BELGIAN POT AZALEA: INTERACTION 

BETWEEN GENETICS, PHYSIOLOGY AND 

CULTURE CONDITIONS 

Pot azalea (R. simsii hybrids) production in Belgium accounts approximately 40 million 

plants per year. The past decade, growers and their associations have made efforts to 

improve both the quality of the vegetative as well as the flowering plants; however, 

suboptimal flowering is often observed. The non uniform opening of flower buds at 

anthesis or flowers that do not entirely open at the consumers place are often observed. 

Problems related to flowering are detrimental for the good image of azalea as a quality 

product. Different potential causes have been quoted: year round production schemes, 

shortening of culture time, increased frequency of application of growth regulators, use 

of more persistent growth regulators or application of assimilation lights when forcing the 

plants. However, a clear cut direct cause is seldom found; interaction between several 

elements related to the culture conditions should be at the base. 

The main objective of this research project is to identify influential factors related to 

flowering quality by an integrated approach focusing on the induction of processes at the 

level of RNA expression, on plant physiology and/or on experimental variation in culture 

conditions. In its practical implementation, we will start with the isolation of homeologous 

candidate genes for regulation of flowering, which will be transformed to expression 

markers using qPCR. Besides, relevant physiological, biochemical and morphogenetic 

parameters will be developed. Application of plant growth regulators to induce the 

transition from vegetative to generative growth under controlled cultured conditions is 

supposed to interfere with the later flowering process: need for cold, dormancy breaking 

of the flower buds, anthesis during forcing of the plants in the greenhouse and open 

flowering at the consumers place. Focused experiments based on the currently used 

culture conditions by the growers are designed for the 3 critical stages (transition, 

dormancy breaking and anthesis). In a later stage of the project, we want to try to 

validate the results on batches of plants taken from the azalea growing companies that 

display abnormalities in flowering. 

P20 

Ellen De Keyser (1) 

Annelies Christiaens 

(1,2) 

Marie-Christine Van 

Labeke (2) 

Els Pauwels (3) 


Bruno Gobin (3) 











Gent, Belgium 

(3) PCS-Research Centre 

for Ornamental Plants, 

Belgium 

ellen.dekeyser@ilvo. 

vlaanderen.be 

Session Posters 93

P21 

D.P. de Vries (1) 

Lidwien A.M. Dubois (2) 

(1) Present address: 

Foundation Sub Rosa, P.O. 

Box 4097, 6710 EB Ede, 



International, P.O. Box 16, 


Netherlands 


DIFFERENCES IN THE GROWTH AND 

DEVELOPMENT OF PRUNED AND UNPRUNED 

'SONIA' CUT ROSE BUSHES 

4-w-old ‘Sonia’ combination plants, bench grafted onto Rosa canina ‘Inermis’ stocks, 

were planted in 40 L containers in a heated glasshouse and grown from February till 

December. Plants were treated, either as cut rose plants in the traditional plant 

architecture (pruned plants, PP), or were left completely undisturbed (UP). At seven predetermined 

dates, 5 complete plants of each PP or UP category were dug up and the 

(fresh) weights of their shoots and roots recorded. Similarly, all fresh weights of the 

prunings (harvested parts) in the PP category were recorded. Both in absolute (AGR) 

and in relative terms (RGR), the PP plants grew slower than unpruned (UP) ones. The 

total fresh weights of PP plants (excluding the weight of the prunings) was reduced to 

about 40% of those of the UP, but including the weight of the prunings this was only 17% 

of the UP. Pruning affected neither the time of emergence nor the number of bottombreaks, 

but in PP plants the diameter of these shoots was about 30% smaller. Similarly 

their root collars were about 30% thinner. At the end of the experiment the shoot/root 

(fresh) weight ratio of UP plants was about 2.5 times higher than of PP plants. In PP 

plants a functional root/shoot equilibrium of about 5.0 seemed to be established. 

94 Session Posters

INHERITANCE OF 2N GAMETES FORMATION 

IN A F1 AND F2 POPULATION OF BEGONIA 

HYBRIDS 

In Begonia several 2n producing genotypes exist. Among these, Begonia ‘Orococo’, 

produces a high number of first division restitution (FDR) pollen as was shown by the 

formation of mainly diads during microsporogenesis. Crosses with B. ‘Orococo’ as father 

plant and B. soli-mutata, B. ‘Art Hodes’ and B. ‘Orococo’ as mother plants resulted in a 

total of 306 seedlings, of which 295 were triploid and only 11 diploid. In the reciprocal 

cross with B. ‘Orococo’ as mother plant and B. soli-mutata as father plant 50 diploids and 

only 3 triploids were detected, indicating that B. ‘Orococo’ produces also unreduced egg 

cells but in lower frequencies compared to unreduced pollen. In order to study the 

inheritance of 2n pollen formation, both pollen size and microsporogenesis aberrations in 

50 triploid and 29 diploid F1 seedling were investigated. In all triploid seedlings monads 

or diads were observed. In several cases, also a high number of polyads were present. 

However the frequency such pollen was observed varied significantly between individual 

seedling. As a result, all seedlings produced large (2n) or only bad sterile pollen. None of 

them produced only normal pollen without 2n pollen. In contrast, only 13 of the 29 

investigated diploid seedlings produced monads or diads. Six of these seedling 

produced no pollen mother cells at all. As a result, only a part of the diploid seedlings 

produced large 2n pollen, while several other seedlings produced only normal pollen or 

no pollen at all. The germination capacity of the F1 pollen was in general poor, although 

in some cases germination percentages reached above 20%. Some F1 triploids were 

used to produce tetraploid F2 progeny. A total number of 14 seedlings was obtained. Of 

the 8 investigated tetraploids, 6 produced monads or diads. The obtained results show 

that the heritability of 2n pollen production is very high. 

P22 

Angelo Dewitte (1) 

Johan Van 



(2,3) 

(1) KATHO,Dept. HIVB, 

Wilgenstraat 32, 8800 

Roeselare, Belgium 

(2) Inst. for Agricultural and 

Fisheries Research (ILVO), 

Plant Sciences unit, 

Applied Genetics and 



(3) Ghent University, 

Faculty of Bioscience 

Engineering, Dept. for Plant 

Production, Coupure links 


johan.vanhuylenbroeck@ 

ilvo.vlaanderen.be 

Session Posters 95

P23 

Anurag Dhyani 

B.P. Nautiyal 

M.C. Nautiyal 

High Altitude Plant 

Physiology Research 

Centre, Srinagar Garhwal, 

Uttarakhand, INDIA 

anuragdhyani@gmail.com 

PHENOLOGY OF LILIUM POLYPHYLLUM 

D.DON EX ROYLE: IN GARHWAL HIMALAYA, 

INDIA 

Phenology is the study of periodically occurring natural phenomenon and their relation to 

climate and changes in season. Phenological progression in Lilium polyphylum of family 

Liliaceae was observed under natural habitats in temperate region of Garhwal, North 

West Himalaya, India. Considering the endangered status and medicinal values viz., anti 

ageing and aphrodisiac, species is prioritized for conservation and cultivation. However, 

this species shows distinct growth behavior as long gestation period follows short 

reproductive phase. Furthermore, it depends immensely on micro climatic conditions, 

therefore noticeable variation is observed in the commencement of different 

phenophases. 

In nature, seed germinates during June- July after 80 days of dispersal when 

temperature reaches between 15 to 20°C and atmospheric humidity between 60-85%. 

Thereafter, it develops and remains in bulblet stage throughout the growth period (late 

September-October). First true leaf produces after 8 months of germination when winter 

season is over and climatic conditions are suitable during March – April (second year) 

immediately after new spouting. Shoot emerges from bulb during mid march and attain 

maximum height during June with bud maturation. Juvenile phase continues with the 

development of aerial parts as well as bulb. Once juvenile phase achieve its peak, 

reproductive stage instigate and flowering starts during mid June approximately after 5- 

8 years of seed germination. Flowering period remains for 15- 20 days and maximum 

blooming was observed when temperature ranges between 18-20°C with atmospheric 

humidity at 45-50%. Seed setting initiate during August followed by slow ripening of a 

capsule which takes nearly two months to mature. Seed shedding starts from mid 

October and continues till mid November simply by splitting of pods. During entire 

vegetative growth period bulb increases in size and biomass by adding scales and 

accumulation of reserve food material and during every winter, experiences a 

vernalization. Since phenophases are very good indicators of plant response to 

environment (climate in particular) and seasonal timing events can be critical for survival 

of life and reproduction. In general, phenological study provides background to functional 

rhythms of plant communities. Besides, the study will help to determine appropriate 

requirements for germination and seedling establishment for domestication purpose as 

well as will help in determining the best harvesting time for bulbs of the species for 

commercial use. 

96 Session Posters

HAPLOID AND DOUBLED HAPLOID 

PRODUCTION VIA ANTHER CULTURE IN 

GENTIAN 

Gentian has been used as ornamental cut- and pot- flowers in Japan. Many gentian 

cultivars have been established from two species (Gentiana triflora and G. scabra), and 

composed of F 1 hybrid varieties and clonal ones. Although many F 1 hybrid cultivars have 

been developed, it remains the problem that the difficulty to produce homozygous 

parental lines. The gentian is highly heterozygous and exhibits intense inbreeding 

depression by self-pollination. Production of doubled haploids from male and female 

gametophytic cells by in vitro culture has been reported in many crops. However, 

application of this technique to gentians has not been reported. In this study, we report 

successful production of haploids and doubled haploids of gentians by anther culture. 

Embryos were obtained from anthers cultured on the 1/2NLN liquid medium with 

13% sucrose after 2 months of culture. After these embryos were transferred to the 

regeneration medium, they could develop into plantlets. Although many embryos were 

produced in G. triflora, the genotypic variation on embryogenesis was observed among 

genotypes used. Flow cytometric analysis and count of chromosome number revealed 

that regenerated plants consisted of haploids, diploids and triploids, especially triploids 

reached to 70% of them. In G. scabra, a few embryos were produced on anther culture 

and only one plant regenerated. 

These results indicate that anther culture is a useful method to obtain haploids and 

doubled haploids in gentians. Breeding programs and genetic studies using the doubled 

haploids obtained in this study are currently carried out. 

P24 

H. Doi (1) 

R. Takahashi (2) 

T. Hikage (2) 

S. Yokoi (1) 

Y. Takahata (1) 

(1) Faculty of Agriculture, 

Iwate University, Morioka, 

Iwate 020-8550, Japan 

(2) Hachimantai City 

Floricultural Research and 

Development Center, 

Hachimantai, Iwate 028- 

7592, Japan 

u0207019@iwate-u.ac.jp 

Session Posters 97

P25 

Angela Etcheverry 

Maria Alemán 

Trinidad Figueroa 

Carlos Gómez 

Stella Pérez 

Dinca Martín 

Cátedra de Botánica, 

Laboratorio de Biología 

Reproductiva, Facultad de 

Ciencias Naturales, 

Universidad Nacional de 

Salta, Calle Buenos Aires 

177, 4400 Salta, Argentina 

avetcheverry@yahoo.com.ar 

FABACEAE FROM NORTHWESTERN 

ARGENTINA AND THEIR POTENTIAL USE AS 

ORNAMENTALS 

The Fabaceae, one of the largest families of the Angiosperms, are amply distributed in 

both hemispheres, from wet tropics and across temperate zones. Many of them are 

characteristic of disturbed places, demonstrating a good adaptation to grow and to 

reproduce under unfavorable conditions, e.g., soils poor in nitrogen, given their capacity 

to fix the atmospheric nitrogen by symbiosis with species of Rhizobium. Within this 

family, the subfamily Papilionoideae represents 70%, and most of the cultivated species 

of legumes belongs to this group. In addition to the crops used as food, the 

Papilionoideae are used also as ornamentals, as Lathyrus odoratus and Spartium 

junceum. As a first step in the characterization of native species of Papilionoideae from 

Northwestern Argentina, with potential use as ornamentals, we determined the floral 

characteristics and life cycle of the following species: Centrosema virginianum (L.) 

Benth; Cologania ovalifolia H.B.K.; Crotalaria megapotamica Burk.; Crotalaria micans 

Link; Crotalaria pumila Ort.; Crotalaria stipularia Desv.; Desmodium cuneatum Hook & 

Arn.; Desmodium incanum AD.; Desmodium subsericeum Malme; Desmodium 

uncinatum (Jacq.) DC.; Galactia latisiliqua Desv.; Indigofera parodiana Burk.; Indigofera 

suffruticosa Mill.; Macroptilium erythroloma (Benth.) Urban; Macroptilium fraternum; 

Phaseolus vulgaris L. var. aborigeneus (Burk.) Baudet; Rynchosia edulis Griseb.; Vigna 

caracalla (L.) Verdc. and Zornia contorta Mohl. Desmodium spp. have flowers contained 

in racemose inflorescences, they are pink coloured and present colour change toward 

violet-blue. Crotalaria spp, Rynchosia edulis and Zornia contorta have yellow flowers 

and racemose inflorescences. Macroptilium spp. and Indigofera spp. possess orange 

flowers. Cologania ovalifolia flowers are fuchsia, while the flowers of Phaseolus vulgaris 

and Galactia latisiliqua are pink. Centrosema virginianum and Vigna caracalla have 

large flowers, (2-5cm.), and they are violet-coloured. In the studied group, Crotalaria 

micans, Crotalaria megapotamica and Vigna caracalla produce a very intense 

fragrance. Considering the life cycle, 89% of the studied species are perennials. In 

relation to rewards, 72% of species produce nectar and this feature enhances their 

attractiveness because of pollinator’s presence. 

98 Session Posters

CHARACTERIZATION OF VIGNA 

CARACALLA FRAGRANCE 

Vigna caracalla L. Verdc. (Fabaceae: Papilionoideae) produces large and beautiful 

flowers with an intense, pleasant fragrance and due to this fact, this species could 

be grown as an ornamental. The objective of this initial study was to identify, using 

headspace analysis, the major volatiles present in the floral bouquet of cut flowers. 

The eluted volatiles were analyzed using a gas chromatograph/mass spectrometer. 

Chromatographic separation was achieved using a PE-Wax column (50 m, 0.25 

mm, 0.25 um). The predominant class of compounds was identified as 

monoterpene hydrocarbons which on average accounted for over 47% of the total 

volatiles. Within this group, the predominating acyclic compounds accounted for 

41% of the total volatiles. Major monoterpene hydrocarbons were identified as 

trans-ocimen, α-farnesen and linalool. The former compound has previously been 

reported as the main constituent of floral scents from other leguminous flowers 

including sweet pea (Lathyrus odoratus) and faba beans (Vicia faba). One acyclic 

monoterpene alcohol (linalool) and one cyclic monoterpene alcohol (indol) 

accounted 21.6% of the total volatiles detected. The former compound has been 

detected in the floral scents of a wide range of plant species including alfalfa and 

sweat pea. A number of previously reported aromatic compounds were detected 

and together accounted for on average 21% of the total headspace. These included 

aldehydes, alcohols and esters, with phenethylalcohol being consistently the most 

abundant. Aromatic compounds have previously been shown to be present at high 

concentrations in the floral bouquet of other leguminous flowers such as white and 

red clovers. From this study it is clear that, as expected, the volatile profile of V. 

Caracalla flowers consists of a complex mixture, with a total of 32 compounds 

being detected in quantifiable amounts. 

P26 

Angela V. Etcheverry (1) 

Stefan Vogel (2) 

María M. Alemán (1) 

Trinidad Figueroa Fleming (1) 

Carlos Gómez (1) 

(1) Cátedra de Botánica, 

Laboratorio de Biología 

Reproductiva, Facultad de 

Ciencias Naturales, Universidad 

Nacional de Salta, Calle Buenos 

Aires 177, 4400 Salta, Argentina 

(2) Institut für Botanik und 

Botanischer Garten der Universität 

Wien, Rennweg 14, A-1030, Wien 

angelaetcheverry@salnet.com.ar 

Session Posters 99

P27 

Tom Eeckhaut 

Johan Van 

Huylenbroeck 

ILVO, Plant sciences Unit, 

Plant Applied Genetics and 



tom.eeckhaut@ilvo. 

vlaanderen.be 

OPTIMIZATION OF CHRYSANTHEMUM 

PROTOPLAST CULTURE FOR ASYMMETRIC 

HYBRIDIZATION 

Asymmetric (somatic) hybridization is an alternative for sexual hybridization and 

symmetric protoplast fusion. The latter two have to be followed by repeated 

backcrossing to limit the introgression of undesired genes, whereas asymmetric 

hybridization may instantly yield end products. However, both genome fragmentation of 

a donor parent and protoplast regeneration of an acceptor parent may create bottlenecks 

for the generation of such hybrids. The aim of this research was to optimize several 

parameters for successful culture of protoplasts of a selection of suitable receptor 

genotypes of Dendranthema grandiflora. 

Altogether, 5 cultivars were selected. The effect of several parameters on both initial 

and sustained development (respectively yielding microcolonies and microcalli) was 

studied. As well growth conditions, media contents and culture systems were evaluated. 

More specifically, incubation time, enzyme mixture, agarose concentration, the evolution 

of the osmotic gradient, complex N-sources, ammonium content, sucrose level, 

presence of organic acids, the presence of nurse tissue and phytohormonal levels were 

the parameters whose effect was monitored. 

Protoplast isolation could be optimised by very mild (10 rpm) overnight (16h) shaking 

in the following enzyme mixture: 0.5% cellulase, 0.3% macerase and 0.1% driselase. 

Initial protoplast divison was accomplished in NH 4 free medium enriched with both 

auxins and cytokinins and 20g/l sucrose, based on MS and supplemented with Nitsch 

vitamins. Sustained development up to microcalli (visible to the naked eye) was not yet 

observed. 

Currently, experiments are ongoing in order to improve gaseous exchange in the 

culture containers. This would enhance the oxygen supply to the dividing protoplasts. 

Microcolonies would then have a higher chance to persist their division and yield 

microcalli. This appears to be the critical step in the regeneration process of 

chrysanthemum protoplasts. Bypassing this barriers would thus mean a major step 

forward in the development of asymmetric hybrids in this ornamental. 

100 Session Posters

BACTERIAL WILT OF PELARGONIUM: 

DEVELOPMENT OF A SCREENING-METHOD 

FOR RESISTANCE 

P28 

Pelargonium is one of the most important ornamental plants in home and garden. 

Cultivars are propagated vegetatively. Two bacterial pathogens (Xanthomomas hortorum 

pv. pelargonii, Ralstonia solanacearum) cause bacterial wilt and blight resulting in high 

economic losses. The first symptom of both diseases is the characteristic wilting of 

single leaves. After the invasion of the bacteria, infected stems become brown or black 

and the whole plant is dying. Symptomatically, these two bacterial diseases can not be 

distinguished but a microbiological differentiation is possible. 

As the inoculation pathway is different for these species an aim of this project was to 

develop reliable inoculation methods for both pathogens. These were developed on plant 

material provided by Elsner pac ® Dresden using bacterial strains of the collection of the 

Institute of Resistance Research and Stress Tolerance. 

For the inoculation with X. hortorum pv. pelargonii, contaminated scissors were used, 

because in praxis the disease transmission occurs when preparing cuttings of plants 

during the propagation process. As the natural inoculation by R. solanacearum is via 

roots, the bacteria suspension was filled into pots and the roots of respective plants were 

cutted (wounded) with a knife to improve the bacteria invasion. 

132 genotypes of Pelargonium were tested by both methods in the greenhouse. As a 

result, two genotypes could be identified as highly tolerant to Xanthomonas and one 

genotype as resistant against both pathogens. The resistant genotype will be used in the 

resistance breeding. 

Josefine Engel (1) 

Martin Geibel (2) 

Klaus Richter (1) 

(1) Julius Kühn-Institut, 

Federal Research Centre 

for Cultivated Plants 

Institute for Resistance 

Research and Stress 

Tolerance, Erwin-Baur- 

Straße 27, 06484 

Quedlinburg, Germany 

(2) Elsner pac ® 

Jungpflanzen GbR, 

Kipsdorfer Straße 146, 

01279 Dresden, Germany 

josefine.engel@jki.bund.de 

Session Posters 101

P29 

Yuichi Futagami (1) 

Hiroki Shiota (2) 

Saori Kama (2) 

Katsumi Nakashima (3) 

Ikuo Nakamura (4) 

Masahiro Mii (4) 

Juntaro Kato (2) 

Syoichi Ichihashi (2) 

(1) Graduate School of 

Education, Aichi University 

of Education, Igaya, Kariya, 

Aichi 448-8542, JAPAN 


Aichi University of 

Education, Igaya, Kariya, 

Aichi 448-8542, JAPAN 

(3) Tokai Municipal 

Agricultural Center, 

Fukishima, Tokai, Aichi 

476-0011, JAPAN 





JAPAN 

jkatoh@auecc.aichiedu.ac.jp 

OCCURRENCE OF PROGENIES WITH 

UNEXPECTED DNA CONTENTS OBTAINED 

FROM THE CROSSES USING UNREDUCED 

GAMETE-PRODUCING DIPLOID CULTIVARS 

WITH ONE F GENOME OF CYMBIDIUM 

FLORIBUNDUM (FF GENOME) WITH DIPLOID 

CYM. EBURNEUM 

In the breeding of Cymbidium, interspecific hybridization has frequently been conducted 

to produce novel cultivars. Although most interspecific hybrids did not show the hybrid 

sterility in Cymbidium, diploid cultivars, which have one genome of Cym. floribundum (FF 

genome), designated as diplo-1F cultivar, frequently formed unreduced gamete and 

yielded triploid and tetraploid progenies in the crosses with diploid and tetraploid partner 

plants, respectively. Since a large variation in DNA content between diploid Cymbidium 

species was detected in our preliminary study, DNA contents of unreduced gamete 

inherited were estimated from that of progenies. Flow cytometric analysis of DNA 

contents revealed that most progenies obtained by the crosses using diplo-1F cultivars 

as female parents were triploid or tetraploid depend on the ploidy level of partner plants. 

Moreover, calculated DNA contents of unreduced gametes produced were not exactly 

but nearly the same as the diplo-1F cultivars used as the parent. These results suggest 

that unreduced gametes of diplo-1F cultivars were FDR type. 

In the present study, 10 progenies with tetraploid or over tetraploid DNA contents 

were obtained from the crosses of diplo-1F cultivars as a female parent with the pollen of 

diploid Cym. eburneum. In these plants, calculated DNA contents inherited from diplo-1F 

cultivars were far from that of FDR 2n-gamete. These results suggest that they were 

formed by natural chromosome-doubling of diploid embryos fertilized between n-gamete 

of diplo-1F cultivar and the haploid pollen of Cym. eburneum or by the fertilization 

between SDR 2n-gamete of diplo-1F cultivar and 2n-pollen of Cym. eburneum. 

102 Session Posters

BIODIVERSITY CONSERVATION STRATEGIES 

OF RARE AND ENDANGERED LIGURIAN 

SPECIES 

Ligurian botanical biodiversity is threatened by increasing urbanization of natural 

habitats. Efforts have been done to acknowledge the EU Community COUNCIL 

DIRECTIVE 92/43/EEC on the conservation of natural habitats and of wild fauna and 

flora (Natura 2000). However, little is known about morphophysiological and genetic 

variation in wild populations. The genus Limonium (Plumbaginaceae) has a worldwide 

distribution, with the largest number of species found in the Western Mediterranean 

basin. It includes species with a wide range of ploidies (mostly di-, tri-, and tetraploids) 

and reproductive systems (sexual and asexual through apomyxis), as well as a high 

proportion of hybrid taxa.. Many of these species inhabit salt-rich soils and their range 

has been reduced due to human pressure, resulting in increased isolation. In Italy, 

Limonium avei (De Not.) Brullo et Erben has been found only in a few Sicilian and 

Sardinian areas and one Ligurian site. Erysimum burnati Vidal (Cruciferae) is an 

endemic species of South-Western Alps. There are just a few populations in the Ligurian 

Alps, between 1400 and 1600 m a.s.l.. We are involved in the effort to preserve L. avei 

and E. burnati through ex-situ conservation methods. Tissue culture techniques have 

proved to be good and efficient methods for conservation of threatened species, 

because many plantlets can be obtained from a minimum quantity of original plant 

material and with low impact on wild populations. We have developed micropropagation 

and regeneration protocols for these two infrequent Ligurian plant species. We are 

developing strategies to assess the level of morphophysiological and molecular 

biodiversity among populations and to study the genetic variation within and among 

natural populations. 

P30 

Annalisa Giovannini (1) 

Laura De Benedetti (1) 

Marco Ballardini (1) 

Antonio Mercuri (1) 

Simonetta Peccenini (2) 

(1) C.R.A. - Research Unit 


Ornamental Species, Corso 

Inglesi 508, I-18038, 

Sanremo (IM), Italy 

(2) DIPTERIS – Genoa 

University, Corso Dogali 

1/M I-16136 Genoa, Italy 

annalisa.giovannini@ 

entecra.it 

Session Posters 103

P31 

J.A. Greppi 

J.C. Hagiwara 

Institute of Floriculture, 

CNIA – INTA, De Los 

Reseros y Las Cabañas 

s/nº (1686), Hurlingham, 

Buenos Aires, Argentine 

jgreppi@cnia.inta.gov.ar 

INTERSPECIFIC HYBRIDIZATION AND 

POLYPLOIDY IN MECARDONIA 

Mecardonia Ruiz & Pav., of recent introduction in the ornament plants world market, is 

an American genre of Plantaginaceae (ex Scrophulariaceae) whose distribution covers 

an area from the east of the United States of North America to the north Patagonia and 

Central Chile (Roussow, 1987). It has around 9 herbal species, generally yellow flowers, 

five of which grow in Argentina (Greppi & Hagiwara, inéd.). 

The commercial varieties found at present come from two species M. tenella (Cham. 

& Schltdl.) Pennell from Brasil and Argentina and M. acuminata (Walt.) Small from 

EE.UU. 

Nevertheless, among the original argentinian species, besides the M. tenella, we can 

find some others of great ornamental value, such as M. flagellaris (Cham. & Schltdl.) 

Rossow and M. reneeae Greppi & Hagiwara. 

The former, even tough is not so proper its plant in a flowerpot due to the fact that it 

has a postrate habit, with long stems, it is of ornamental interest, because it naturally 

displays a variability in the colour of its flowers, and they all can be yellow, rose or purple 

and white. On the other side, the M. renneae, has the particularity of being an upright 

and compact plant, and has flowers, though only yellow, bigger than the rest of the 

species of the genre. Also, M.tenella, is an upright or upward plant, with little yellow 

flowers, but with a greater degree of compactness than M. reneeae. 

Given the morphological variability that these species display, it is of great interest 

the obtaining of interespecific hybrids. For that reason, a great number of crossbreeding 

between M. tenella and M. flagellaris and between M.tenella and M. reneeae were did. 

In the first crossbreeding, viable seeds were obtained, which germinated normally, 

while in the second crossbreeding, no seed was obtained. 

Also and so as to obtain a clue of the ploidy level of these species, the same were 

analyzed with a flow cytometer. The outcomes showed different levels of ploidy, being 

M. tenella and M. flagellaris diploids and M. reneeae tetraploids. Given this results, it 

was decide to diploid, by means of the treatment with colchicine, to M. tenella. 

Eventually, poliploids invididuals of M. tenella were crossbreeded with M. reneeae, 

obtaining this time positive outcomes. The hybrids showed characteristics of both 

parents, and some of them result of ornamental interest. 

104 Session Posters

MORPHOLOGICAL AND TISSUE CULTURE 

STUDIES OF PLATYCERIUM CORONARIUM, A 

RARE ORNAMENTAL FERN SPECIES FROM 

MALAYSIA 

The genus Platycerium consists of about 18 species, commonly found in tropical and 

subtropical forest. Among the species found in Peninsular Malaysia are Platycerium 

coronarium, P. platylobium, P. ridleyi and P. wallichi. The most attractive is the the 

majestic P. coronarium. Platycerium coronarium is a gigantic, epiphytic fern native to 

tropical areas of South America, Africa, Southeast Asia, Australia and New Guinea. They 

nest on the upper branches of the tallest tree in the forest. Due to their unique-shaped 

fronds they are popular as ornamental plants which can be found in gardens, especially 

tropical gardens. 

Records on detailed morphological studies of the species are scanty therefore it is 

one of the aims of the study to investigate the macro- and micro-morphological 

characteristics of the species. Studies were carried out on both intact and in vitro plants. 

Scanning electron microscope study revealed the presence of multicellular trichomes on 

the abaxial surface of both intact and in vitro leaves. Sunken, anomocytic-type stomata 

were also observed on the abaxial surface of the leaves. Propagation of the species 

through tissue culture using Murashige and Skoog (1962) media supplemented with 

hormones were used to obtain an efficient regeneration system as well as an approach 

for conservation. The most responsive regeneration of sporophyte leaves was obtained 

when the explants were cultured on MS medium supplemented with 0.1 and 1.5mg/l GA 3 

and 30mg/l sucrose , at pH 5.6 in 16 hours light and 8 hours in the dark. 

P32 

Noorma Wati Hareon 

Rosna Mattaha 


Sciences, Faculty of 


Malaya, 50603 Kuala 

Lumpur, Malaysia 

noorma@um.edu.my 

Session Posters 105

P33 

Rohollah Heidarhaee (1) 

Alireza Babaei (2) 

Reza Amiri (3) 

Shirin Dianati (1) 


Horticultural Science, 

College of Aburaihan, 

University of Tehran, 

Tehran, Iran 


Horticultural Science, 

Tarbiat Modares University 

(TMU), P.O. Box: 14115- 

365, Tehran, Iran 


Agronomy, College of 

Aburaihan, University of 

Tehran, Tehran, Iran 

arbabaei@modares.ac.ir 

A PRELIMINARY STUDY ON GENETIC 

DIVERSITY OF THE IRANIAN IRISES 

Iris is known as an important ornamental plant and Iran has been mentioned as one of 

its origins. A study has been carried out in order to analyze the genetic variation of 

Iranian Iris species. Initially, RAPD markers were used to determine the genetic diversity 

level and phylogenetic relationships among 16 genotypes of Iris species including wild 

material. A total of 14 random primers were used, 12 of which showed good amplification 

and polymorphism and the combination of these primers was found optimal for 

discrimination of the genotypes with very low values of cumulative confusion probability. 

Overall, 722 bands were produced, and 680 bands were polymorphic. Unweighted pair 

group method cluster analysis based on Jaccard’s similarity values revealed 2 and 7 

groups at the distance of 0.23 and 0.35 respectively. Preliminary results showed a broad 

genetic diversity among Iranian Irises. We consider a more intensive sampling across 

the western and northern parts of the country and using a more precise molecular 

marker in order to find a better understanding of Iranian Irises biodiversity for further 

analyses. 

106 Session Posters

BREEDING WOODY ORNAMENTALS AT APR, 

LISSE 

Applied Plant Research in Lisse has a long tradition of breeding ornamental nursery 

stock. The breeding of woody plants is characterised by the long generation time of the 

plants, which often take 7 years or more to flower for the first time after sowing. Some 

recent cultivars were the result of over 20 years of breeding and selection. 

Pieris japonica ‘Passion’ is an evergreen shrub, characterised by its cherry red 

flowers in upright inflorescences. In 1980 P.j. ‘Cupido’ was crossed with P.j. ‘Valley 

Rose’. The F1 had upright inflorescences, but all white flowers. Crossing the F1 with P.j. 

‘Valley Valentine’ in 1988 finally resulted in the introduction of ‘Passion’ in 2008. 

Female Skimmia japonica s.str. can have nice red berries, but since it is dioecious, it 

requires a male pollinator plant. Plants of S. japonica subsp. reevesiana are 

monoecious, so self-pollinators do occur in this species. In 1989 several S. japonica 

s.str. cultivars were crossed, and their offspring was screened for female plants which 

also had functioning anthers. After propagation and tests for cultivation properties, one 

monoecious selection will be introduced in 2009 as S. japonica ‘Temptation’. It can self 

pollinate to produce berries, but still has the abundant bright red berries and good health 

typical of S. japonica s.str. 

Hypericum x inodorum is a subshrub that suddenly became very popular for cutting 

in the 1990’s. Both the cut branches and the garden plants of this hybrid are very 

susceptible to rust (Melampsora hypericorum). We screened many wild accessions of 

the hybrid’s parent species H. androsaemum and H. hircinum for rust resistance. We 

crossed resistant plants with H. x inodorum ‘Excellent Flair’ in 1995, and in the offspring 

we selected a compact rust-resistant plant with an abundance of large berries. It was 

introduced in 2004 as H. x inodorum ‘Arcadia’. It is excellent for containers, gardens and 

landscaping. 

P34 

Margareth E.C.M. Hop 

Wageningen University and 

Research centre, Applied 

Plant Research 


margareth.hop@wur.nl 

Session Posters 107

P35 


Dong Hee Suh (1) 

Song Kyoung Park (1) 

Aung Htay Naing (1) 

In Suk Park (1), 

Si Yong Kang (2) 

In Suk Ahn (1) 





702-701, Korea 

(2) Dept. of Radiation Plant 

Breeding and Genetics, 

Advanced Radiation 



Institute, Geongeup 580- 

185, Korea 


KARYOTYPE ANALYSIS OF LILIUM SPECIES 

BELONGING TO MARTAGON SECTION BY 

FISH TECHNIQUE 

The genus Lilium which is consisting of over 100 species belongs to the family Liliaceae, 

and is mainly distributed in Northern Hemisphere. The genus Lilium is divided into seven 

sections, Martagon, Pseudolirium, Lilium, Archelirion, Sinomartagon, Leucolirion and 

Dauroliruon. The basic chromosome number of genus Lilium has been known as 

2n=2x=24, with the exception of triploid (2n=3x=36) species, L. lancifolium. Martagon 

section consists of five species, such as L. martagon, L. hansonii, L. tsingtauense, L. 

distichum, and L. medeoloides. L. hansonii is one of the Korean native lily species, and 

originated from Ulleung island of Korea. L. tsingtauense is known as the only up-facing 

Martagon lily species and shows resistance against Botrytis. L. martagon has abundant 

flowers with dark purple-red coloured flowers and vigorous growth. L. distichum has 

orange-red coloured flowers and dark spot. L. medeoloides which is known as “wheel 

lily” is a common lily species in Japan. Fluorescence in situ hybridization (FISH), using 

45S and 5S ribosomal DNA, clearly revealed nucleolar organizing region (NOR) and 

complements of each chromosome. We have constructed detailed karyotypes of L. 

hansonii (2n=2x=24), L. tsingtauense (2n=2x=24), and L. distichum (2x=2x=24). Seven 

pairs of 45S rDNA signals were in L. hansonii, four pairs in L. tsingtauense, and four 

pairs in L. distichum. Only one pair of 5S rDNA signal was detected in all speices of L. 

hansonii, L. tsingtauense, and L. distichum. The results clearly demonstrate that 

Martagon section could be discriminated from other Lilium taxa through karyotype 

analysis. 

108 Session Posters

KARYOTYPE ANALYSIS OF SEVERAL 

SUBGENUS ROSA SPECIES BY FISH 

TECHNIQUE 

Rose is one of the most economically important ornamental crops in the world, due to its 

utlilization as pot or cut flowers and landscape shrubs. The genus Rosa is taxonomically 

divided into four subgenera, including Hulthemia, Platyhodon, Hesperhodos, and Rosa. 

Subgenus Rosa is comprised of 10 sections including Pimpinellifoliae, Rosa, Caninae, 

Carolinae, Cinnamomeae, Synstylae, Indicae, Banksianae, Laevigatae, and Bracteatae. 

The basic chromosome number of them is seven and their ploidy level ranges from 

diploid (2n=2x=14) to octoploid (2n=8x=56). Because of its small chromosome size, 

cytogenetic analyses of genus Rosa were rarely performed until now, and similar 

chromosome morphology makes it difficult to discriminate their homologous 

complement. Fluorescence in situ hybridization (FISH) technique is an ideal method for 

discriminating chromosomes each other by observing specific markers, such as 

ribosomal DNA repeat. In this study, we have analyzed karyotypes of seven wild rose 

species, R. multiflora, R. rubus, R. soulieana, R. chinensis, R. mulligani, R. indica, and 

R. gallica, based on the chromosome length and FISH signals which are the result of 

hybridization with 45S ribosomal DNA. The number of somatic chromosomes was 

2n=2x=14 in R. multiflora, R. rubus, R. soulieana, R. mulligani, R. indica, 2n=4x=28 in R. 

gallica, and 2n=6x=42 in R. chinensis. Two FISH signals of 45S rDNA were observed in 

diploid species of R. multiflora, R. rubus, R. soulieana, R. mulligani, and R. indica, four 

signals in R. gallica, and six signals in R. chinensis. All 45S rDNA was positioned on the 

terminal region of short arm of chromosomes in seven wild rose species. In conclusion, it 

was feasible to identify the genus Rosa by karyotype and physical mapping analyzed 

using ribosomal DNA. 

P36 


Won Hee Kim (2) 

Sung Tae Kim (2) 

Youn Yol Han (3) 

Min Kyung Kwon (3) 

Kiu Weon Kim (4) 

Jae-Dong Chung (1) 





702-701, Korea 

(2) National Inst.of 

Horticultural & Herbal 

Science, Suwon-si, 

Gyeonggi-do, 441-440, 

Korea 

(3) Gumi Floricultural 

Experiment Station, 

Gyungbuk Provincial ATA, 

Gumi 130-831, Korea 

(4) Dept. of Horticultural 

Science, Yeungnam 

University, Dae-dong 214- 

1, Gyeongsan 712-740, 

Korea 


Session Posters 109

P37 

Vahid Jajarmi-Islamic 

Azad University Bojnord 

Branch-IRAN 

Vahid_jajarmii@yahoo.com 

THE EFFECTOF IBA, NAA HORMONE ON 

STIMULATING ROOT GENERATION IN MALVA 

CHINESIS 

To study the effect of indole boteric acid(IBA) and naftalin ascetic acid(NAA) hormones 

on stimulating root germination in MALVA CHINESIS ,an experiment was carried out 

four replication through factorial in completely randomized. The hormones were of 

0,1000,2000,3000 mgr/lit . Concentrations cutting of 1-1.4 centimeter diameter were put 

in the solutions for 60 seconds. Then they were put in three cultural media of gravel, 

sand, soil with decomposed manure. The results showed that in 3000 mgr/lit of IBA 

concentration more roots were generated(8.2). Though there was no statistical 

difference between 2000 and 3000 mgr/lit concentration. The longest root was seen in 

2000 mgr/lit concentration of IBA and NAA. The most drought weight of root was 

obtained in 3000 mgr/lit concentration of IBA. The most percentage of root producing 

belonged to 2000 mgr/lit NAA ..The best cultural media were gravel and sand. 

110 Session Posters

INITIATION OF ENDOPOLYPLOIDY IN SEED 

DEVELOPMENT OF PHALAENOPSIS 

APHRODITE SUBSP. FORMOSANA AND ITS 

APPLICATION TO POLYPLOID BREEDING 

Endopolyploidy is the occurrence of cells with different ploidy in the same plant tissue as 

a result of chromosome DNA duplication without cell division. It is commonly found in the 

mature tissues of many plant species, including Phalaenopsis orchids. The objective of 

this study was to locate the critical stage in the development and germination of seeds 

as well as the position of the embryonic tissues by which the endopolyploidy was 

initiated in Phalaenopsis aphrodite subsp. formosana. The methods for this study 

included a combination of the analysis of DNA content by flow-cytometry and the 

cytological study by DAPI staining and fluorescence microscope equipped with Zeiss 

ApoTome Slider. It was shown that the growth of ovaries and the development of ovules 

began after pollination. By 43-50 days after pollination (DAP), 2C nuclei in the embryonic 

tissues were dominant over 4C nuclei. In addition, meiosis and fertilization took place at 

this stage. During seed development (50-85 DAP), 4C nuclei increased rapidly and 

became dominant over 2C nuclei as a result of intensive mitosis. After 90 DAP, 2C 

nuclei began to increase and reached the plateau level by 120 DAP. Seeds became 

mature and dormancy began. Endopolyploid cells with 4C nuclei occurred at the stage of 

embryo development. After sowing of seeds, 8C nuclei was observed 4 days after 

sowing (DAS) indicating the existence of endopolyploidy at the very early development 

stage of of protocorms. The optical sections of the protocorms between 10-30 DAS 

observed by using the Zeiss ApoTome Slider showed that large and prominent nuclei 

occurred at the basal region of the protocorms. These large nuclei increased as the DAS 

increased and was concurrent with the increase in 8C nuclei in the protocorm tissue at 

this stage. In this study, endopolyploidy in Phalaenopsis aphrodite subsp. formosana 

was found to occur in the embryonic tissues of the seeds before and right after sowing. 

The application of this finding to polyploid breeding in the orchid is discussed. 

P38 

Goamg-Tyng Jean 

Yu-Lin Kao 

Ching-Yan Tang 

Wen-Huei Chen 

Institute of Biotechnolgy , 

National University of 

Kaohsiung, Kaohsiung 811, 

Taiwan, R.O.C. 

a08539@nuk.edu.tw 

Session Posters 111

P39 

Sladjana Jevremovic 

Angelina Subotic 

Milana Trifunovic 

Marija Petric 

Institute for Biological 

Research “Sinisa 

Stankovic”, Bulevar 

despota Stefana 142, 

11060 Belgrade, Serbia 

sladja@ibiss.bg.ac.yu 

FLOWERING OF DWARF IRISES DERIVED BY 

TISSUE CULTURE 

Plant regeneration by somatic embryogenesis and/or organogenesis has been achieved 

by zygotic embryo culture of two dwarf irises (Iris pumila and I. reichenbachii). Both 

regeneration processes are induced on MS nutritional media supplemented with only 2, 

4-dihlorophenoxy acetic acid (2, 4-D). Concomitant somatic embryogenesis and 

organogenesis are achieved at concentration of 0.1-10 M 2, 4-D for I. pumila and 0.5- 

1.0 M for I. reichenbachii. Embryogenic calus is separated and cultured on medium 

with 2, 4-D and kinetin (0.5 and 5.0 M, respectively) where somatic embryos are 

formed. Germination of somatic embryos (70 %) is achieved on MS hormone free 

medium. At the surface of organogenic calli the shoots are formed. Afterwards, shoots 

are cultured on MS media for shoot multiplication supplemented with BAP and GA 3 (0.1 

and 0.3 M, respectively) where some plants flowered. In vitro formed flowers of I. 

pumila have the same colour and morphology as donor plants. Fully regenerated 

plantlets with both processes acclimatized in next flowering season. Plants of I. 

reichenbachii with changed shape and number of flower parts are observed. Altered 

phenotype was manly present among plants that are regenerated by organogenesis 

following described protocol. Some of I. reichenbachii plants have doubled all flower 

parts, which can be very useful for further improvement of irises since the investigated 

species are main parents for breeding of dwarf irises. Detail analysis of regenerants is in 

progress. 

112 Session Posters

ASSESSMENT OF TULIPS (TULIPA L.) 

VEGETATIVE REPRODUCTION POTENTIAL 

The principal aim of the research is assess vegetative reproduction potential of the 

whole mother bulbs cross-section according to special reproduction coefficient. 

Numerous investigations on vegetative reproduction potential of tulips have been 

carried out worldwide, but until now more thorough studies have been fulfilled with 

comparatively little number of cultivars. 

Vegetative reproduction capacity of the whole mother bulb spectrum (7 fractions) of 

299 cultivars was ascertained according to special reproduction coefficients: total 

reproduction coefficient (TRC), generative bulb reproduction coefficient (GRC) and 

forcible bulb reproduction coefficient (FRC). Reproduction coefficients were calculated 

individually for each studied fraction of the investigated tulip cultivars. TRC is a 

quantitative indicator, specifying mean number of all daughter bulbs per clone. GRC is a 

qualitative indicator, specifying mean number of capable to blossom next year bulbs per 

clone. FRC is a qualitative indicator, specifying mean number of forcible tulip bulbs per 

clone. By modulating the data on TRC, GRC and FRC of the whole mother bulb crosssection, 

indexed reproduction coefficient (IRC) was deduced. IRC indicates comparative 

reproduction value of the whole mother bulb cross-section of the studied tulip cultivars. 

Empirical tulip cultivar dispersion analysis demonstrated that this coefficient most 

objectively reflects reproduction capacity of all fraction bulbs of the studied tulip cultivars. 

Basing on IRC, the investigated tulip cultivars were grouped into 5 grades of 

reproduction. According to IRC, most tulip cultivars were attached to 2nd – 4th grades 

(correspondingly 24, 30 and 30 %), whereas 8 % of the studied cultivars occurred in 1st 

and 5th grades. 

P40 

R.Juodkaitė (1) 

A.Baliūnienė (2) 

G.Stukėnienė (1) 

S.Dapkūnienė (1) 

S.Jančys (3) 

S.Žilinskaitė (1) 

A.Skridaila (1) 

(1) Botanical Garden of 

Vilnius University, Kairėnų 

43, LT-10239, Vilnius, 

Lithuania 

(2) Field Floriculture 

Research Station, A. 

Kojelavičiaus 1, LT-2048, 

Vilnius, Lithuania 

(3) Institute of Botany, 

Žaliųjų ežerų 49, LT-2021, 


regina.juodkaite@ 

gmail.com 

Session Posters 113

P41 

Z. Ghayoor Karimiani (1) 

G.H. Davarynejad (2) 

A. Bagheri (1) 

M. Jafarkhani Kermani 

(3) 

(1) . Department of Agrobiotechnology, 

Ferdowsi 

University of Mashhad, Iran 

(2) . Department of 

Horticulture, Ferdowsi 

University of Mashhad, Iran 

davarynej@um.ac.ir 

(3) . Agricultural 


Institute of Karaj, Iran 

EFFECT OF ORYZALIN ON CHROMOSOME 

DOUBLING AND IN VITRO GROWTH OF 

GERBERA JAMESONII 

The possibility of in vitro tetraploid induction via oryzalin treatments and effect of this 

herbicide on growth in Gerbera (Gerbera jamesonii.) was studied. To assay the initial 

ploidy level of “Red Explosion” cultivar, the mother plants were determined by DNA Flow 

Cytometer. Five levels of oryzalin concentration (0, 30, 60, 120 and 240 μM) in modified 

liquid Murashigue and Skoog medium were used at various time of exposing (12, 24 and 

48 hours) on shaker with 92 rpm. Then each plantlet divided into multiplication medium 

with 2 - 4 and 6 mg/lit kinetin. 10 plantlets per treatments were used as replication 

(random factor). The experiment was inducted as factorial based on completely 

randomized design for chromosome doubling and growth comparison. In order to peruse 

the effect of oryzalin on growth, fresh weight and number of leaves in days 22 nd and 44 th 

after oryzalin treatments were measured. Also 12 weeks after oryzalin treatments, ploidy 

level was evaluated by DNA Flow Cytometer. Minimum increase in fresh weight (0.17 g) 

was obtained in treatment containing 240 μM oryzalin, 2 mgL -1 kinetin and 48 hours 

exposure period. The lowest number of new leaves was produced on the treatment of 

120 µM oryzalin for 24h on the medium containing 4mgL -1 kinetin whereas the minimum 

fresh weight was obtained on the treatment of 240 µM oryzalin for 48h on the medium 

containing 4mgL -1 kinetin. There was no observation of tetraploid plants in lower 

concentration (30 and 60 μM oryzalin) but in these treatments chimeras were obtained. 

Maximum percentage of tetraploid plant was recorded at 240 and 120 μM oryzalin 

treatments in 48 hours. The results show that however only in higher level of oryzalin 

concentration the tetraploid tissues can be obtained, effect of this antimitotic agent on 

growth is significant. 

114 Session Posters

EVALUATION OF PRE-FERTILIZATION 

BARRIERS BY OBSERVATION OF POLLEN 

TUBE GROWTH AND ATTEMPTS FOR 

OVERCOMING POST-FERTILIZATION 

BARRIERS IN INTERGENERIC 

HYBRIDIZATION BETWEEN ALSTROEMERIA 

AND BOMAREA BY OVULE CULTURE 

The genus Alstroemeria is a continuous demand for new cultivars as an ornamental 

flower. Many interspecific hybrids of Alstroemeria are utilized for commercial cultivars. 

However there is no report about intergeneric hybridization. For introducing novel 

characters into Alstroemeria, Bomarea species, which is related genus to Alstoroemeria, 

is selected in this study. We are investigating the possibility of creating intergeneric 

hybrids between Alstroemeria and Bomarea. Previously, we found that there was a postfertilization 

barrier between A. aurea and B. coccinea. In the present study, we 

investigated species-dependant differences of the frequency of pollen tube entry into 

ovules after intergeneric pollination. Moreover, ovule culture conditions were examined 

for overcoming post-fertilization barriers after pollination of Bomarea pollen grains to A. 

aurea, A. pelegrina var. rosea and A. magenta. 

The frequency of pollen tube entry into ovules were compared in the pistil of A. 

aurea, A. pelegrina var. rosea and A. magenta after pollination of B. coccinea pollen 

grains. Pollen tubes in Alstroemeria ovaries were observed with aniline blue staining 48 

hours after pollination. The frequencies of pollen tube entry into ovules were 0.3%, 5.6% 

and 10.0% in A. aurea, A. pelegrina var. rosea and A. magenta, respectively. For the 

ovule culture, pollinated ovaries were harvested 3 and 7 days after cross pollination and 

the ovules were cultured on 2 g l -1 gellan gum-solidified MS medium with or without 

gibberellic acid and supplemented with sucrose at different concentrations (30, 60, 80 or 

100 g l -1 ). As a result, 3 plantlets were obtained in A. pelegrina var. rosea × B. coccinea 

cultured on MS medium supplemented with 80 g l -1 sucrose. 

Although pollen tubes of B. coccinea were reached to A. aurea ovules, this 

combination might have stronger pre-fertilization barriers than those of A. pelegrina var. 

rosea and A. magenta. It supposed to be different intensity of pre-fertilization barriers 

among species. Our date also suggested that sucrose concentration at 80 g l -1 in culture 

medium was effective to obtain progenies after intergeneric pollination. Confirmation of 

hybrid natures for the plantlets is now in progress. 

P42 

Yukiko Kashihara (1) 

Tomonari Hirano (2) 

Naho Murata (3) 

Koichi Shinoda (3) 

Hajime Araki (1,4) 

Yoichiro Hoshino (1,2,4) 

(1) Division of Biosphere 

Science, Graduated School 

of Environmental Science, 

Hokkaido University, Japan 

(2) Division of Innovative 

Research, Creative 

Research Initiative 

‘Sousei’(CRIS), Hokkaido 

University 

(3) National Agricultural 

Research Center for 

Hokkaido Region, Japan 

(4) Field Science Center for 

Northern Biosphere, 

Hokkaido University, Kita 

11, Nishi 10, Kita-ku, 

Sapporo 060-0811, Japan 

ikuy06@exfarm.agr. 

hokudai.ac.jp 

Session Posters 115

P43 

Juntaro KATO(1) 

Mayuko IKEDA (2) 

Mai HAYASHI (3) 

Rieko ISHIKAWA(1) 

Yasuko YOSHIDA (4) 

Ryo OHSAWA (4) 

Kei-ichiro MISHIBA (5) 

Norio TANAKA (6) 

Ikuo NAKAMURA (3) 

Masahiro MII (3) 


Aichi University of 

Education, Hirosawa, 

Kariya, Aichi 448-8542, 

JAPAN 


Bioagricultural Sciences, 

Nagoya University, 

Morowa, Tougou, Aichi 

470-0151, JAPAN 





JAPAN 




Tsukuba, 1-1-1 Tennodai, 

Tsukuba Ibaraki 305-8572, 

JAPAN 



Sciences, Osaka Prefecture 

University, 1-1, Gakuen, 

Sakai, Osaka 599-8531, 

JAPAN 

(6) Tsukuba Botanical 

Garden, National Museum 

of Nature and Science, 4-1- 

1 Amakubo, Tsukuba, 

Ibaraki 305-0005, JAPAN 

jkatoh@auecc.aichiedu.ac.jp 

DIFFERENCES IN PLOIDY LEVELS AMONG 

INTERSPECIFIC HYBRIDS OBTAINED FROM 

THE CROSS COMBINATIONS USING PRIMULA 

SIEBOLDII AS FEMALE PARENT 

Primula sieboldii (Section Cortusoides) is one of the traditional ornamental plants in 

Japan. In cultivars of P. sieboldii, 63 diploids, 8 triploids and 1 tetraploid were reported 

by somatic chromosome observation (Yamaguchi 1973). 

Flow cytometric analysis of 200 cultivars also revealed that present population of 

cultivars in P. sieboldii was consisted with diploids, triploids and tetraploids. When we 

conducted interspecific crosses using P. sieboldii as a female parent, different patterns 

of ploidy levels, which were categorized into three types, were found among progenies 

according to interspecific cross combinations. 1) Both many diploids and few triploids 

were included in the same cross combinations, 2) Either diploids or triploids were 

obtained depending on the female cultivars of P. sieboldii, 3) All hybrids are triploid. 

Types 1, 2 and 3 were found from the intersectional wide crosses with the pollen of P. 

obconica (Section Obconicolisteri), from the intrasectional crosses with the pollen of P. 

jesoana (Sect. Cortusoides) and from the intrasectional crosses with the pollen of 

Primula kisoana (Sect. Cortusoides), respectively. Differences in contribution of 

unreduced gametes for fertilization and/or embryogenesis were discussed in relation to 

the interspecific cross combinations of P. sieboldii with different species as male parents. 

116 Session Posters

MOLECULAR CYTOGENETIC ANALYSIS OF 

UNILATERAL AND BILATERAL SEXUAL 

POLYPLOIDIZATION IN RELATION TO 

INTERGENOMIC RECOMBINATION AND 

INTROGRESSION IN LILIUM SPECIES 

HYBRIDS 

Longiflorum (L), Asiatic (A) and Oriental (O) lilies belong to section Leucolirion, 

Sinomartagon and Archelirion of genus Lilium respectively. These interspecific hybrids 

(LA and OA) are promising in lily breeding for various agronomical traits. Both LA and 

OA hybrids produce 2n gametes and have been used to develop sexual polyploids by 

backcrossing to Asiatic parents as well as by sib-mating of the F1 LA hybrids. The BC1 

progenies were triploid, with few exceptions and the progenies from sib-mating were 

tetraploid or near tetraploids. Genomic in situ hybridization (GISH) technique was 

applied to assess the intergenomic recombination in the BC1 populations of LA and OA 

hybrids obtained after unilateral sexual polyploidization. It was found that in LA hybrids, 

LA × AA and in reciprocal crosses (AA × LA) plants were originated through the 

functioning of either 2n eggs or 2n pollen. Similarly the BC1 OA hybrids comprised of 

triploid plants which originated through functional 2n pollen from a diploid OA hybrid. In 

both type of crosses, a majority of the progenies originated through First Division 

Restitution (FDR) mechanism of functional 2n gametes either with or without a cross 

over with few exceptions where Indeterminate Meiotic Restitution (IMR) was the 

mechanism of 2n gamete formation. Based on GISH analyses it was found that most of 

the LA and OA hybrids exhibited recombination. Intergenomic recombination was also 

estimated in the progeny of sib-mated LA hybrid. In this case both parents had 

contributed gametes with the somatic number of chromosomes (i.e., 2n-2n). This 

population originated through bilateral sexual polyploidization. These allotriploid 

interspecific lily hybrids with recombinant chromosomes obtained from unilateral sexual 

polyploidization can be used for the selection of desirable traits at triploid level. However, 

allotetraploids obtained from bilateral sexual polyploidization can be fertile and used as 

parents repeatedly to produce triploid or tetraploid progenies. 

P44 

Nadeem Khan 

M.S. Ramanna 

Richard G.F. Visser 

Jaap M. van Tuyl 

Laboratory of Plant 

Breeding, Wageningen UR, 

Droevendaalsesteeg 1, 

6708 PB Wageningen, The 

Netherland 

Jaap.vantuyl@wur.nl 

Session Posters 117

P45 

Kamal S. Kirad (1) 

Swati Barche (2) 

Morseng Modi (3) 

D.B. Singh (3) 

(1) Department of Welfare 

& Agril. Devlopment, 

Tikamgarh, Govt of MP, 

India 

(2) JNKVV, Jabalpur , 

College of Agriculture, 

Tikamgarh, M.P. India 

(3) Allahabad Agricultural 

Institute-Deemed 

University, Allahabad, U.P, 

India 

kskal24@rediffmail.com 

GENETIC VARIABILITY IN CHRYSANTHEMUM 

A huge quantum of variability exists in chrysanthemum with respect to shape, size, 

growth habit, flowering behavior, vase life etc. The phenotypic variability observed in 

chrysanthemum is further partitioned into different heritable and non-heritable 

component of variation with suitable genetic parameters such as genetic coefficient of 

variation (GCV), heritability, genetic advance, correlation and path analysis. The main 

emphasis in the present investigation was therefore, given to study the genetic variability 

in chrysanthemum which can be used in chrysanthemum improvement programme. An 

experiment was conducted at research farm, Department of Horticulture, Allahabad 

Agricultural Institute-Deemed University, Allahabad during winter season of 2006-07. 

Twenty genotypes of chrysanthemum were used for variability studies and replicated 

thrice. High genotypic coefficients of variation (GCV) and phenotypic coefficient of 

variation (PCV) estimates were found for yield of flower per hectare followed by duration 

of flowering and vase life. High heritability with high genetic advance was observed for 

yield of flower per hectare, duration of flowering and flower diameter. Positive and direct 

correlation of flower diameter with flower yield reveal that flowers production can be 

increased by directly selecting flowers with more diameter. 

118 Session Posters

HYDRANGEA SUSPENSION CULTURE AS 

SOURCE FOR STABLE PROTOPLASTS 

Hydrangeas as garden shrubs but also as potted plants and for floristry enjoy a truthful 

renaissance. There are more than 90 species of the genus Hydrangea. Most originate 

from Asia, some of them from the American continent. It could be assumed that this 

plentiful gene pool will be used for the development of new varieties, although up to now 

in breeding processes the use of biotechnological methods for hydrangeas is rare. 

A protocol for the development and maintenance of suspension culture for 

Hydrangea quercifolia was elaborated. For primary callus induction explants from 

petioles were cultivated on ½ B5 medium supplemented with 10 g L -1 sucrose, 15 g L- 1 

glucose, 4 mg L -1 NAA and 0.09 mg L -1 TDZ. After about 5 weeks callus was transferred 

to a liquid medium with the same compounds as for the induction. The cultures were 

cultivated in the dark on a shaker with 100 rpm. All liquid media were changed by fresh 

ones every week. After two months without visible growth, the callus slices began to 

proliferate to round lumps. After a further three months the clusters were crushed using a 

spatula and transferred to liquid MS medium with 40 g L -1 sucrose and 1 mg L -1 2,4-D. In 

that medium fine suspension was developed. Now, the subculture of the suspension will 

be performed by pipetting a small amount of suspension cells to fresh medium every 

week. The microscopic observation shows smooth cell clusters with many divisions. 

Therefore, the suspension cells could be useful as an unlimited source for protoplasts. 

P46 

Evelyn Klocke 

Simone Abel 

Julius Kühn Institute (JKI), 

Federal Research Centre 

for Cultivated Plants, 

Institute for Breeding 

Research on Horticultural 

and Fruit Crops, Erwin- 

Baur-Str. 27, D-06484 


evelyn.klocke@jki.bund.de 

Session Posters 119

P47 

Nobuo Kobayashi 

Mika Ishihara 

Masanori Ohtani 

Kyeong Seong Cheon 

Daiki Mizuta 

Keisuke Tasaki 

Akira Nakatsuka 

Faculty of Life and 

Environmental Science, 

Shimane University, 

Matsue, Shimane, 

690-8504 Japan 

nkobayashi@life.shimaneu.ac.jp 

EVALUATION AND APPLICATION OF FLOWER 

LONG-LASTING TRAIT (MISOME-SHO) OF 

AZALEA CULTIVARS 

The long-lasting trait of flower corolla with temporal change of colour exists in Japanese 

evergreen azalea. This trait called misome-sho has been found out in several species 

and cultivar groups from Edo era. For the purpose of application of this trait for breeding, 

we investigated several characteristics and gene expression of these cultivars. 

Wild species of Rhododendron kaempferi, R. indicum and R. macrosepalum and 

flower long-lasting cultivars of each species were used in this study. 

All flower long-lasting cultivars have smaller corolla compared to wild types and have 

stomata in corolla surface, while wild types do not have stomata in their surface. Pot 

planted flower long-lasting cultivars kept their corolla more than 100 days and corolla 

colour was changed from red or purple-red in anthesis to green in later. In contrast, wild 

type plants finished flowering in about 20 days. In microscopic observation of longitudinal 

section of flower, abscission layer in basal portion existed in wild type corolla are not 

observed in flower long-lasting cultivars. 

Expression of two class B MADS genes: AP3- type and PI-type genes isolated from 

Kurume azalea was analyzed. AP3- type gene express in whole 2, 3 and 4 

corresponding to corolla, stamen and pistil organ of wild species, but do not express in 

any whole of flower long-lasting cultivars. The long-lasting trait of corolla would be 

derived from sepaloid petal caused by homeotic gene mutation. 

These cultivars have high pollen fertility and stabilized seed set ability. Inheritance of 

this trait is researching using cross progenies. 

120 Session Posters

ENDANGERED ORNAMENTAL SPECIES 

Recently estimation shows that about 3900 ornamental species (e.g. 13.9% of total 

ornamental species) are under threat and there is a weak positive correlation between 

number of threatened ornamental species and total number of species in related 

families. According to IUCN categories ” Endangered species ” are those that taxa in 

danger of extinction and whose survival is unlikely if the causal factors continue 

operating. Included are taxa whose numbers have been reduced to a critical level or 

whose habitats have been so drastically reduced that they are deemed to be in 

immediate danger of extinction. The application of this category was done for ornamental 

species using Geln’s book, Cultivated Plants of Southern Africa and IUCN’s red list of 

plants. Totally 92 endangered species is recorded. The highest number of endangered 

species found in the family of Palmae (21) followed by Bromiliaceae (16), Zamiaceae 

(14) and Cupressaceae (6). Most of species were presented here is used only for 

ornamental purposes but some have multi-purposes. For example Juglans hindsii 

(Jepson) Jepson ex R. E. Sm. (Cupressaceae), that is frequently used as rootstock for J. 

regia L. because of its resistance to disease. Another multi-purposes plant is Malus 

hupehensis (Pamp.) Rehd. (Rosaceae) that provide excellent resistant to scab and 

mildew and is used as rootstock for apples. From the roughly 20 species of endangered 

crops species already listed by Hammer and Khoshbakht in 2005, only 2 also appear 

here in the endangered ornamental category (ca. 10%). The overlapping between the 

groups of ornamental and crop plants will be discussed here. 

P48 

Korous Khoshbakht (1) 

Karl Hammer (2) 

(1) Shahid Beheshti 

University; G.C., 

Environmental Science 

Research Institute, Tehran, 

Iran 

(2) University of Kassel, 

FB11, Institue of Crop 

Science. Steinstr. 19, D- 

37213 Witzenhausen, 

Germany 

k-khoshb@sbu.ac.ir 

Session Posters 121

P49 

Edgar Krieger 

CIOPORA, Gänsemarkt 45, 

20354 Hamburg, Germany 

info@ciopora.org 

STRENGTHENING BREEDER’S 

INTELLECTUAL PROPERTY PROTECTION 

WITH THE CONCEPT OF EDV – CIOPORA’S 

POSITION 

Many misunderstandings still exist, when it comes to the concept of EDV. CIOPORA 

believes that only a clear interpretation of the EDV concept will provide fair and 

predictable solutions on this matter and protect the intellectual property of innovative 

breeders properly. In this regard the CIOPORA position paper distinguishes between two 

types of EDV: varieties which are solely based on the genome of the initial variety and 

where the genomic structure is highly conserved, e.g. spontaneous and induced 

mutants, GMO and apomicts on one side and “Me-too-varieties” on the other. For both 

groups CIOPORA developed clear rules. 

The concept of "Essentially Derived Varieties" is a mixture of technical 

(describing) and legal aspects. It is a true extension of the breeders´ right and a 

temporary limitation of the breeders´ exemption. Taking into account Article 14 (5) (b) of 

the UPOV 1991 Act CIOPORA is of the opinion that an asexually reproduced ornamental 

and fruit variety shall be deemed to be essentially derived from another variety (the initial 

variety) if it 

a) is clearly distinguishable from the initial variety, 

b) is predominantly derived from the initial variety or from a variety that is itself 

predominantly derived from the initial variety and 

c) except for the differences which result from the act of derivation, conforms to the 

initial variety in the expression of the essential characteristics that result from the 

genotype or combination of genotypes of the initial variety. 

We explain how to deal with these prerequisites and with legal disputes in this 

respect. 

For further details please find attached the CIOPORA position paper on EDV and the 

cover letter. 

122 Session Posters

GENETIC VARIABILITY IN DAVIDIA 

INVOLUCRATA SPECIMENS GROWING IN 

POMERANIA 

The aim of conducted studies was to determine genetic and morphological variability of 

selected Davidia involucrata specimens from Western Pomerania and Berlin using ISSR- 

PCR technique. 

The studies were carried out on the Davidia involucrata var. ”Vilmoriniana” growing in 

Germany in the Botanical Garden in Berlin-Dahlen and in Poland, in Pomerania, nearby 

the Polish-German border: in the Dendrological Garden in Przelewice, Glinna and 

Central Cemetary in Szczecin. 

ISSR technique made it possible to determine genetic variability of the examined 

specimens. This was done by means of 6 out of 30 ISSR primers used in the 

experiment. Six primers (802, 807, 810, 819, 839, 840) generated 64 visible amplification 

products, of which 11 were: monomorphic, 31 – polymorphic, 12 - genotype specific. On 

average 1 primer generated 10 amplification products which ranged from 2550bp to 

266bp.The greatest number of ISSR-PCR products was observed in the range of 2550- 

266bp. 

The analysis of phylogenetic tree showed that there was 60-87% genetic similarity 

between the examined specimens. The similarity between the youngest and the oldest 

tree in Przelewice Arboretum was 86,6%: between them and the davidia from Central 

Cemetary in Szczecin - 80% and 69.4% between the genotypes from Berlin and Glinna. 

Low genetic variability between the genotype from Glinna, Berlin, Central Cemetary and 

Przelewice may result from the fact that they are probably the descendants of the trees 

from a pre-war German nursery in Berlin. The davidia from Kornik, originated from 

English nurseries and introduced to Poland in 1931, turned out to be the most genotype 

specific . 

P50 

Danuta Kulpa 

Miłosz Smolik 

Marianna 

Dobrochowska 

Agricultural University, ul. 

Janosika 8, 71-424 

Szczecin, Poland 

danuta.kulpa@agro.ar. 

szczecin.pl 

Session Posters 123

P51 

Ja-Hyun Lee (1) 

Ye-Sun Chung (1) 


Geung-Joo Lee (2) 


Yong-Kweon Yoo (3) 

Tae-Ho Han (1) 


Horticulture, Division of 

Plant Biotechnology, 

Chonnam National 

University, Gwangju 500- 

757, Korea 

(2) Advanced Radiation 



Institute, Jeongeup 580- 

185, Korea 

(3) Major in Horticultural 

Science, Division of 

Bioscience, Mokpo National 

University, Muan 534-729, 

Korea 


INDUCTION OF MUTATIONS IN 

CHRYSANTHEMUM (DENDRANTHEMA 

GRANDIFLORA) BY USING GAMMA-RAY 

IRRADIATION 

Chrysanthemum (Dendranthema grandiflora) is one of the important cut flowers all over 

the world as well as Korea. Dendranthema grandiflora cv. Beakma is a white standardtype 

cultivar developed in Korea and produce high quality cut flowers throughout the 

year. However, this cultivar shows vacant space in stem during high temperature in 

summer. The hardness of its stem is weakened and hard to handle by warping and 

bending. 

The objective of this study was to induce mutant without vacant space of stem for 

improvement of quality in Dendranthema grandiflora cv. Beakma. In addition, the effect 

of radiation was investigated on the survival and growth rate of individuals. A total of 

1,679 rooted cuttings were treated with different doses of gamma rays (10~50 Gy). All 

individuals survived regardless of irradiation-dose but the growth was decreased with an 

increase of radiation-dose. Particularly, plant height and internode length were 

remarkable diminished from 2 to 4 times at 40 and 50 Gy. In morphological 

characteristics of leaves, leaf length and width were increasingly decreased, and petiole 

length was increased as increase of dose. 

In contrast the respiratory quotient was significantly increased about 3 times at 50 Gy 

in comparison with the control. The irradiated individuals were repeatedly propagated 

and an individual was selected as a mutant without vacant space inside the stem in a 

total of 7,109 individual stems. Cuttings from this individual were rooted for propagation 

and another selection. 

We conclude that treatment of gamma-ray may be an effective way for inducing 

exclusive mutation of Dendranthema grandiflora. We will analyze by using RAPD 

markers and propagate from induced mutants in the future. 

124 Session Posters

STUDY ON HEREDITY RULES OF CYCLAMEN 

PERSICUM COLOURS 

To identify the theoretic foundation for maintaining and improving high quality varieties 

and breeding new F1 hybrids, flower colour segregation occurred in the offspring in both 

self-pollination and cross-pollination of varieties of different colours were investigated. 

The main results indicate that the heredity of Cyclamen persicum colour follows both 

quantitative and qualitative hereditary regularity. All genes can be classified as two 

groups of the throat-gene and coronal-gene occupying different spots on the 

chromosome. The coloured gene is the dominant gene, while the white gene is the 

recessive one. The gene groups and their quantity determine the colour of cyclamen 

persicum. And sometimes the throat- gene and the coronal-gene take a recombination, 

forming a new type of flower. 

P52 

Lin, Yan (1) 

Yin, Shulian (2) 

BI Jun (1) 

Wang, Chunrong (1) 

Guo, Weizhen (1) 

(1) Hebei Academy of 

Forestry science, 

Shijiazhuang 050061, 

P.R.China 

(2) Department of garden in 

Hebei Professional College 

of Political Science and 

Law, Shijiazhuang 

050061,P.R.China 

linyan65@yahoo.com.cn 

Session Posters 125

P53 

Levko G.D. 

All-Russian Institute of 

Vegetable Breeding and 

Seed Production, 

Moscow region, Odintsovo 

district, p/o Lesnoy 

Gorodok, 143080, Russia 

flowers@vniissok.ru 

INHERITANCE OF FLOWER COLOURS IN 

SWEET PEAS (LATHYRUS ODORATUS L.) 

As a result of biochemical studying structure of pigments and character of inheritance of 

flower colours at eight varieties, used in system diallelic crossings, and also at ten 

hybrids F1 of a sweet peas (Lathyrus odoratus L.) it has been established, that this 

attribute is controlled, at least, by four basic genes synthesizing anthocyanin pigments 

and three inhibitor- genes of these pigments. 

126 Session Posters

HEAT TOLERANCE IN PETUNIA AS 

MEASURED BY AN ELECTROLYTE LEAKAGE 

TECHNIQUE 

Identification of heat-tolerant petunia (Petunia ×hybrida Hort. Vilm. Andr.) genotypes and 

techniques for rapid assessment of heat-tolerant plants during breeding programs for 

sub-tropical and tropical areas are desirable. The extent to which electrolyte leakage 

from petunia leaf discs at 50 °C for 20 min, measured using a test for cell membrane 

thermostability (CMT), could be related to the reduction in branch number induced by 

heat in the greenhouse-grown plants were determined. Heat-intolerant cultivars exhibited 

more reduced branches, but less reduced RI (relative injury) value than heat-tolerant 

cultivars with increasing mean growing temperatures from 16 to 27 °C. The cultivars with 

a high RI value are those with the lesser CMT and more reduction in branch number by 

high temperature at 27 °C. The relationship between the RI value occurring in leaf tissue 

discs of two seed-propagated cultivars and treatment temperature from 25 to 56 °C was 

sigmoidal. The RI values at the approximate midpoint of the sigmoid response curve 

occurred at 47 °C for ‘Primetime Carmine’ regardless of growing day/night temperatures, 

and at 47 and 49 °C for ‘Tidal Wave Silver’ grown at 25/20 °C and 30/25 °C, 

respectively. A high temperature at 30/25 °C resulted in reduced branch number in 

‘Primetime Carmine’ but not in ‘Tidal Wave Silver’. 

P54 

Tzu-Hsuan Liu 

Chia-Yang Lin 

Der-Ming Yeh 

Department of Horticulture, 

National Taiwan University, 

No. 1, Roosevelt Road 

Section 4, Taipei, 106, 

Taiwan 

dmyeh@ntu.edu.tw 

Session Posters 127

P55 

Supuk Mahadtanapuk 

(1) 

Mondhon 

Sanguansermsri (2) 

Rataporn Chandej (3) 

Somboon 

Anuntalabhochai (4) 






Thailand 

(2) Faculty of 

Pharmaceutical Sciences, 

Naresuan University, 

Phitsanulok 65000, 

Thailand 

(3) Dept. of Biology, Faculty 

of Science, Maejo 



(4) Dept. of Biology, Faculty 

of Science, Chiang Mai 




ISOLATION AND EXPRESSION ANALYSIS OF 

A GENE ENCODING ACC OXIDASE IN 

CURCUMA ALISMATIFOLIA GAGNEP 

To understand the molecular functions of ACC oxidase genes in curcuma and to mean 

ethylene production knockout by recombinant technique, cloning and expression of ACC 

oxidase genes are essential for anti-senesce or gene silencing technique that reduces 

ethylene production, ultimately enhancing the storage life and quality of the harvested 

products. In this study, cDNA fragments encoding ACC oxidase from Curcuma 

alismatifolia Gagnep. were isolated and its expression analyzed. Highly conserved 

Primers were designed from ACC oxidase’s various plants of GenBank database. The 

forward and reverse primers were derived from ENWGFFE and TNGKYKS amino acid 

segment, respectively. The PCR products had length of 600 basepair and were 

subcloned into pGEM T-easy vector resulting in pCa-ACO1. After sequencing, the 

deduced amino acid sequence of the cDNA was highly homologouse to those of ACC 

oxidase gene isolated from other plant in NCBI database. The expression of the gene 

during postharvesting, organ different and wounding was investigated. Northern blot 

analysis shows that Ca-ACO1 gene is expressed at petal and bract of curcuma and high 

accumulated at 1 day in petal and 3 days in bract at postharvesting of curcuma. This 

result was related the respiration and the ethylene production of open florets increased 

as they approached senescence of the curcuma flower. We suggest that a Ca-ACO1 is 

one of ACC oxidase genes that relate in ethylene production and senescence in petal 

and bract of curcuma after harvesting. 

128 Session Posters

HYBRID STATUS OF ‘ELIATOR’-BEGONIAS 

ANALYSED BY GISH 

Interspecific hybridization of various tuberous begonia species hybrids with Begonia 

socotrana results in so-called ‘Eliator’- begonias hybrids. In our study, karyotypical 

differences between parental genotypes was recorded regarding chromosome size and 

number. Somatic complement of B. socotrana comprised of 28 short chromosomes with 

a length of about 0.5 μm (2n = 28) whereas tuberous begonia had 52 chromosomes of a 

size ranging from 2 to 3 μm. A number of ‘Eliator’- begonias hybrids were analysed by 

genomic in situ hybridization (GISH) and flow cytometry for their mode of origin. For 

GISH, genomic DNA of tuberous begonia was sonicated to 1–10-kb fragments, labeled 

by nick translation with digoxigenin-11-dUTP and was used as a probe and B. socotrana 

DNA was autoclaved to 100 bp fragments and used as block. In hybrids two groups 

comprises short and long chromosomes were recorded. The long chromosomes of 

tuberous begonia were uniformly labelled green and their number ranged from 14 to 65 

depending on the hybrids whereas the number of short B. socotrana chromosomes 

equaled 14. In case of two genotypes tested no B. socotrana chromosomes were 

recorded in putative hybrids. Recombination between genomes of tuberous begonia and 

B. socotrana was not observed. In addition, their hybridity was readily verified by flow 

cytometry. Thus, genomic in situ hybridization and flow cytometry analyses can be useful 

to identify the genome constitution of ‘Eliator’- begonia hybrids and thus gain an insight 

into the origins of these cultivars. 

P56 



M.S. Ramanna (2) 

Wendy ter Laak (3) 








(2) Wageningen University 

& Research Center, Plant 

Breeding, Wageningen, 


(3) Beekenkamp Plants 

B.V., Maasdijk, the 

Netherlands 

agnieszkamarasek@wp.pl 

Session Posters 129

P57 

Y. Matsushita (1) 

K. Sumitomo (1) 

Y. Chikuo (1) 

M. Shibata (2) 

(1) Research Team for 

Growth and Flowering, 

National Institute of 

Floricultural Science, 2-1, 

Fujimoto, Tsukuba 305- 

8519, Japan 

(2) Agriculture,Forestry and 

Fisheries Research 

Council, 1-2-1, 

Kasumigaseki, Chiyoda-ku, 

Tokyo 100-8950, Japan 

yousuken@affrc.go.jp 

INVESTIGATION OF THE GENETIC 

RESISTANCE OF CHRYSANTHEMUM 

MORIFOLIUM TO CHRYSANTHEMUM STUNT 

VIROID 

Stunting caused by chrysanthemum stunt viroid (CSVd) is one of the most damaging 

diseases of cultivated chrysanthemum (Chrysanthemum morifolium), the most important 

cut flower in Japan. This disease has been reported in many regions of the world. The 

symptoms are severe: stunting of plant height, reduction in flower size, and bleaching of 

the flower. It is difficult to cultivate viroid-free chrysanthemum plants, and CSVd-resistant 

cultivars have not yet been reported. The objective of our research was to find CSVdresistant 

cultivars and to investigate their inheritance pattern. We screened 40 

chrysanthemum cultivars for resistance to CSVd. Scions of the screened cultivars were 

inoculated with CSVd by grafting them onto CSVd-infected plant roots. Two months after 

the grafting inoculation, we analyzed the upper leaves of the scions by reverse 

transcription polymerase chain reaction (RT-PCR) in order to detect CSVd, according to 

the method of Hosokawa et al. (2005). CSVd infection occurred in all the cultivars except 

for ‘Okayamaheiwa’, which did not show any symptoms of infection. F 1 progenies were 

produced by crossing the resistant cultivar ‘Okayamaheiwa’ with a susceptible cultivar, 

namely, ‘Sei-elza’. From approximately 200 seeds obtained from the F 1 progenies, 17 

were randomly selected and grown; their seedlings were inoculated with CSVd by 

grafting. Of the 17 F 1 progenies inoculated with CSVd, 5 were not infected with CSVd, 

suggesting that the characteristic of resistance is inherited in C. morifolium. 

130 Session Posters

FLOWER COLOUR AND PIGMENT 

COMPOSITION OF DIFFERENT ORCHID 

HYBRIDS 

Orchid industry ia a multibillion-dollar business and it has become an important 

contributor to the country’s economy. Orchid flower colour is an important factor in 

determining the marketability of a particular hybrid. There is a pressure to produce 

hybrids with new flower colour to meet the everchanging customers preferences. In 

order to achieve these objectives there is a need to understand the linkage between 

flower colour and the types of pigments involved. The information obtained is important 

in developing research strategies especially in selecting the right hybrids for colour 

manipulation and breeding purposes. A study, therefore, was carried out to establish the 

relationship between flower colour and pigment composition of different orchid hybrids. 

Flowers with colours ranging from white and shades of pink, orange and blue were 

studied. Different pigments which include anthocyanins, carotenoids and chlorophyll of 

the flower petals were analysed. The hybrids studied were Mokara Chark Kuan Orange, 

Mokara Chark Kuan Pink, Aranda Chark Kuan Blue, Dendrobium Sonia 17, D. Savin 

White, Vanda Mimi Palmer, Vanda white, Phalaenopsis bellina, P. bellina var alba, 

Oncidium Sharry baby and Oncidium Taka. The pigments of new hybrid D. Alya Pink 

and its parents (D. Tengku Anis x D. bigibbum) were also analysed. Generally the 

colourful hybrids D. Sonia 17, V. Mimi palmer, P. bellina and O. Sharry baby contain 

significant amount of anthocyanins as compared to white types P. bellina var alba, D. 

Sonia 17 and V White. Oncidium Taka, which is predominantly yellow in colour was 

found to contain a small quantity of anthocyanin but its β-carotene content is seven-fold 

higher than that of anthocyanin content. Three major anthocyanins were identified in the 

flower extracts. They were Petunidin, Delphinidin and Malvidin. The major anthocyanins 

in D. Sonia 17 is Petunidin and Malvidin, O. Sharry Baby and V. Mimi Palmer contain 

Delphinidin while Petunidin and Delphinidin are found in P. bellina. A small amount of 

anthocyanin was detected in the yellow and white flowers. Chlorophyll is present in all 

the hybrids including the white types. There is no clear variation in the pH values of the 

flower sap although there appears to have a lower pH value in the white types. 

P58 

Mahmood Maziah 

Azzreena Mod Azzeme 

Suhaili Shamsi 

Nisha Nambiar 

Department of 

Biochemistry, Faculty 

Biotechnology and 

Biomolecular Sciences. 

Universiti Putra Malaysia. 

43400. Serdang. Selangor 

malaysia.aziahm@biotech. 

upm. edu.my 

Session Posters 131

P59 

J. Meiners (1) 

T. Winkelmann (2) 

(1) Research Station for 

Horticulture, 

Weihenstephan University 

for Applied Sciences, Am 

Staudengarten 8, 85354 

Freising, Germany 

(2) Leibniz University of 

Hannover, Institute of 

Floriculture and Woody 

Plant Science, 

Herrenhaeuser Str. 2, 

30419 Hannover, Germany 

julia.meiners@fhweihenstephan.de 

OVULE CULTURE OF HELLEBORUS SPECIES 

Within the genus Helleborus about 21 species are found, part of which has been used 

for production of interspecific hybrids with new interesting characteristics. However, 

other interspecific combinations could result in new ornamentals and would be of 

commercial as well as scientific interest. In order to establish protocols for embryo 

rescue techniques, in a first step this study aimed at the identification of culture 

conditions for ovules from intraspecific crosses. The effects of dissection date after 

pollination and temperature during ovule culture were examined in intraspecific crossings 

of Helleborus niger, H. argutifolius, H. x hybridus and H. foetidus. For these species the 

period for seed ripening under natural conditions takes ten to twelve weeks depending 

on the climate. 

Ovaries were harvested three to six weeks after pollination, surface disinfected in 70 

% ethanol for 30 sec, 2 % sodium hypochlorite with one drop Tween for 10 min and 

rinsed in sterilised water three times. Ovules were dissected from ovaries and cultured 

on medium based on MS (Murashige & Skoog, 1962) solidified with 0.4 % Gelrite at a 

pH of 5.8. Two media supplemented with 2.5 or 5 % sucrose were compared. Ovules 

were cultured in darkness at 24 ± 1 °C or 16 ± 1 °C for twelve weeks. Thereafter, ovules 

of each temperature treatment were split and one half was incubated at 6 ± 1 °C for 

eleven weeks while the other half remained in the initial temperature. Afterwards the 

ovules were placed back to their initial temperature. During the following weeks 

germination was evaluated. 

On average of all species, 2.0 % of the ovules dissected three and 0.6 % dissected 

four weeks after pollination germinated, while preparation after five weeks resulted in 3.4 

% and after six weeks in 5.6 % germination, respectively. The intermediate cold 

treatment with 6 °C turned out to be very beneficial for later germination. Furthermore, 

60 % of all germinated ovules were obtained on medium supplemented with 2.5 % 

sucrose, assuming that the sucrose concentration is not a crucial factor. The conditions 

identified in this study have now to be verified, if they also hold true in interspecific 

combinations. 

132 Session Posters

REGENERATION OF MEDICINAL PLANT 

CLITORIA TERNATEA FROM MALAYSIA 

Tissue culture studies of medicinal plant Clitoria ternatea (Butterfly Pea) was carried out 

to investigate the regeneration potential this species in vitro. Various explants from 

aseptic seedling were used such as leaves and stems which were cultured on Driver and 

Kuniyuki Walnut medium (DKW) medium together with different concentrations and 

combinations of hormones such as Napthalene Acetic Acid (NAA), 2,4- 

dichlorophenoxyacetic Acid (2,4-D) and Benzyl Aminopurine (BAP) to achieve 

regeneration. This research also focused on getting coloured callus from the explants 

which has potential for coating technologies. The economic importance of this species 

includes anticonvulsant, antidepressant, indigestion, constipation and arthritis, eye 

ailments, as a cover crop and as an ornamental plant in Malaysia. 

P60 

Normadiha Mohamed 




Sciences 

University of Malaya, 50603 

Kuala Lumpur, Malaysia. 

normadiha@yahoo.com 

Session Posters 133

P61 

Y. Morita 

M. Miotani 

M. Mii 






271-8510, Japan 

mrt_snow_01@yahoo.co.jp 

AGROBACTERIUM-MEDITATED 

TRANSFORMATION OF STOCK (MATTHIOLA 

INCANA) 

Stock (Matthiola incana) is one of the important ornamental plants used as cut flowers 

and pot plants. In stock, insect damage by lepidopteran pests has been a serious 

problem, resulting in quality loss and high pesticide cost. In order to reduce the damage 

by pests, it is necessary to produce insect-resistant plants by introducing an endotoxin 

gene of Bacillus thuringiensis into stock cultivars. To achieve the goal, we established 

transformation methods of stock in this study. To establish a system of Agrobacterium 

tumefaciens-meditated transformation, we introduced β-glucuronidase (GUS) gene into 

stock cultivars. Genetically transformed plants of stock were regenerated after cocultivating 

highly regenerable nodular calli with Agrobacterium tumefaciens strain 

EHA101 (pIG121-Hm) that harbored genes for GUS, hygromycin phosphotransferase 

(hpt) and neomycin phosphotransferase II (nptII). When calli of stock maintained in liquid 

Murashige-Skoog medium (MS) were inoculated with Agrobacterium, frequency of GUSpositive 

calli were increased with sonication and vacuum infiltration treatments. 

Adventitious shoots were regenerated from hygromycin-resistant calli after transfer onto 

agar-solidified MS medium containing sucrose and hygromycin. However, it was difficult 

to induce roots from these adventitious shoots. Transformation of the hygromycinresistant 

calli and shoots were confirmed by histochemical GUS assay, PCR analysis 

and Southern hybridization. We have also established adventitious shoot regeneration 

system from leaf disk by using agar-solidified Woody Plant Medium (WPM) 

supplemented with zeatin. By utilizing this regeneration system, hygromycin-resistant 

shoots were successfully obtained through the same Agrobacterium tumefaciensmediated 

transformation method. Transformation was confirmed by histochemical GUS 

assay, and PCR and Southern analyses. 

134 Session Posters

GYNOGENIC HAPLOID INDUCTION IN 

MIMULUS AURANTIACUS: ESTABLISHMENT 

OF INDUCTION CONDITIONS AND 

CHARACTERIZATION OF REGENERANTS 

ACCORDING TO PLOIDY AND 

HOMOZYGOSITY 

Mimulus aurantiacus Curtis is a promising ornamental plant native from North America. It 

is a self-fertile, hummingbird pollinated perennial shrub with tubular hermaphroditic 

flowers of a wide range of colours. The aim of our study was to establish an efficient 

method for gynogenic haploid induction in M. aurantiacus and to develop a reliable 

marker for homozygosity testing of regenerants. 

In situ gynogenesis was performed using pollination with γ-irradiated pollen (600 Gy) 

and was followed by in vitro embryo rescue 11 to 35 DAP. Placenta attached and 

detached ovules isolated from 506 flowers were inoculated on several growth media. 

They consisted of modified Rangaswamy medium with sucrose concentrations ranging 

from 40 to 120 g l -1 and IAA concentrations ranging from 0.1 to 10 mg l -1 . Of 366 

germinated embryos, ploidy of 189 was analyzed using flow cytometry. Ploidy evaluation 

revealed that 4 were haploids, 165 diploids, 3 triploids, 3 mixoploids, 2 aneuploids and 

12 of undetermined ploidy level. 

In order to identify the nuclear origin of the obtained diploid plantlets, a microsatellite 

marker specific for Mimulus species was developed using the cross-genera approach. 

The marker showed a high degree of polymorphism in Mimulus species and the 

heterozygous nature of most M. aurantiacus cultivars tested. The analysis of 64 diploid 

regenerants from haploid induction experiments revealed that 33 of them were 

homozygous for the locus tested while 31 of them were heterozygous and therefore 

hybrids. 

This is the first report about the successful haploid induction in M. aurantiacus. The 

implementation of the technique could be a valuable tool for M. aurantiacus breeding. 

The developed codominant molecular marker for homozygosity testing is essential for 

Mimulus haploid induction research and applications. Early homozygosity determination 

enables efficient selection of doubled haploids from unwanted heterozyogotes at a very 

early stage of the breeding programme which saves time and funds needed for 

phenotypic evaluations. 

P62 

J. Murovec 

B. Bohanec 

University of Ljubljana, 

Biotechnical Faculty, 

Jamnikarjeva 101, 1000 

Ljubljana, Slovenia 

jana.murovec@bf.uni-lj.si 

Session Posters 135

P63 

R. Naderi 

P. Norouzi 

Dept of horticulture, Faculty 

of agriculture, University of 

Tehran, Karaj, Iran 

rnaderi@ut.ac.ir 

DIRECT BULBLET REGENERATION FROM 

BULB SCALE EXPLANTS OF HYACINTHUS 

ORIENTALIS CV CARNEGIE IN VITRO 

The effect of two plant growth regulator, Benzyl amino purine (BAP) and α- 

Naphthaleneacetic acid (NAA), on direct bulblet regeneration from bulb scale explants of 

Hyacinthus orientalis cv. Carnegie investigated in vitro. With upper scale explants, direct 

bulblet regeneration occur in Murashige & Skoog (MS) solid medium containing 1 mg/L 

NAA and 5 mg/L of BAP. Bulblet regeneration on lower scale explants occur in the 

presence of 0.5 mg/L NAA, only. Twelve weeks after culture regenerated bulblets were 

1 – 1.5 cm in diameter. The maximum root formation observed in ¾ MS medium with 0.3 

mg/L NAA. However, either in upper scale explants and lower scale explants, bulblets 

formed on abaxial side of scale explants. After rooting and adaptation, plants transferred 

to soil. Using this method about 200 – 250 hyacinth plantlet can produce from one bulb 

with 3 – 4 cm in diameter. 

136 Session Posters

ISOLATION OF 3-DEOXYANTHOCYANIDIN 

GLUCOSYLTRANSFERASE GENE FROM 

SINNINGIA CARDINALIS FLOWERS 

Most anthocyanins are synthesized from 3-hydroxylated anthocyanidins, pelargonidin, 

cyanidin and delphinidin, etc. On the other hand, 3-deoxyanthocyanins are rare 

anthocyanin pigments in some plant species. They are pigments that provide orange to 

red colours. However, there have been relatively few studies on how the biosynthetic 

pathway produces 3-deoxyanthocyanins. Especially, there is no report on the 

modification of 3-deoxyanthocyanidin, which is important step to accumulate the 

pigments into vacuoles. In this study, we attempted to identify gene encoding 

glucosyltransferase (GT) for 3-deoxyanthocyanidins from Sinningia cardinalis, 

accumulating 3-deoxyanthocyanins abundantly in their petals. Degenerate primers were 

designed from the plant secondary product glycosyltransferase (PSPG) box, which was 

highly conserved among plant GTs. Five GT candidates, designated as ScGT1 - 5, were 

amplified using S. cardinalis flower cDNA as a template, and full-length cDNA for each 

ScGT fragment was cloned using RACE technology. Phylogenetic analysis showed that 

the deduced amino acid sequences of these ScGTs were classified into the UGT88 

clade containing rose 5, 3-GT and snapdragon chalcone 4’-GT. Recombinant proteins of 

each ScGT candidate produced by E. coli expression system were used to investigate 

GT activity for various 3-deoxy and 3-hydroxyflavonoid substrates. As a result, ScGT5 

could specifically transfer a glucosyl moiety to 3-deoxyanthocyanidin, apigeninidin and 

luteolinidin, but not other flavonoids including 3-hydroxyanthocyanidins, flavonols and 

flavones. Enzymatic properties of ScGT5 were also determined. ScGT5 might be useful 

to modify flower colour by genetic engineering in floricultural plants. 

P64 

Takashi Nakatsuka 

Yuko Kakizaki 

Yoshiko Abe 

Norimoto Shimada 

Masahiro Nishihara 

Iwate Biotechnology 

Research Center, 22-174- 

4, Kitakami, Iwate, 024- 

0003, Japan 

nakatuka@ibrc.or.jp 

Session Posters 137

P65 

Takako Narumi (1,2) 


Tomoya Niki (1) 

Takaaki Nishijima (1) 


Keiichiro Hiratsu (3,4) 

Masaru Ohme-Takagi 

(2) 


(1) National Institute of 

Floricultural Science, 

NARO 


Kagawa University 


Genome-based Biofactory, 

AIST 

(4) Department of Applied 

Chemistry, National 

Defense Academy of Japan 

tnarumi@ag.kagawau.ac.jp 

TRANSGENIC TORENIA EXPRESSING 

CHIMERIC AGAMOUS REPRESSOR EXHIBITS 

SERRATED PETALS AS THOSE INDUCED BY 

CYTOKININ APPLICATION 

Chimeric REpressor gene-Silencing Technology (CRES-T) is an efficient gene silencing 

system in which the chimeric repressors derived from various transcription factors 

dominantly suppress the expression of the respective target genes, and the resultant 

transgenic plants exhibited loss-of-function phenotypes specific for the transcription 

factors even in the presence of redundant transcription factors. 

The homeotic protein AGAMOUS (AG) terminates floral meristem and promotes 

development of stamens and carpels in Arabidopsis. Disruption of its function or 

expression of the chimeric AG repressor (AGSRDX) results in redundant petals, known 

as double flower phenotype. We introduced this chimeric repressor into torenia (Torenia 

fournieri Lind.) to investigate whether CRES-T method is applicable to ornamental 

flowers to increase their horticultural value. Transgenic torenia plants expressing 

AGSRDX showed no redundancy in petal number, but exhibited serration in petal 

margins, anthocyanin accumulation and morphological change in stigma surface, 

formation of extra vascular bundles in petals and styles, and development of ectopic 

trichome-like cells in styles. Anatomical observation of petals and styles revealed that 

these phenotypes are highly similar to those of the treated torenia by a synthetic 

cytokinin analog CPPU in the derangement of vascular bundle. These serrated petals 

and extra vascular bundle were also observed when the chimeric repressors for torenia 

C-function genes TfFAR or TfPLE1 were expressed. These results suggest that the 

morphological change in AGSRDX transgenic torenia plants is induced by the disruption 

of C-function, while the novel phenotypes might be caused by the modification of 

cytokinin-dependent regulation in vascular bundle formation and /or ectopic expression 

of the chimeric repressors in all whorls by CaMV 35S promoter. 

138 Session Posters

EVALUATION OF RESISTANCE OF 

GAULTHERIA TO COLLETOTRICHUM 

GLOEOSPORIOIDES 

The genus Gaultheria consists of about 100 to 200 species. Most are found in North and 

South America, Canada, New Zealand, Australia, Asia and also in Great Britain. 

Gaultheria procumbens L. is an ericaceous perennial and winter hardy shrub and. It 

becomes an important species in Germany which is cultivated and used as an 

ornamental plant in autumn and winter. It is named by the Canadian medical scientist 

and biologist H. Gautier. 

In the last years there are enormous losses up to collapses of the whole crop in 

German plant companies. Lesions on stems and sometimes on leaves, shoot wilting and 

dieback were observed. The causal agent of this disease is Colletotrichum 

gloeosporioides (Penz.) Penz. & Sacc. (teleomorph: Glomerella cingulata (Stonemann) 

Spauld. & H. Schrenk). C. gloeosporioides is an important economical pathogen 

worldwide on legumes, strawberry, blueberry, citrus fruits, coffee, cocoa and Hypericum 

calycinum (St. John's wort). It is seed transferable and survives on plant remainders 

which serve as a source of inoculum. A prevention of the propagation of the pathogen is 

possible but only in the juvenile phase by frequent application of chemical fungicides. In 

older plants the application of fungicides only is preventively, since the fungus is 

colonizing the lower stem part, where fungicides do not arrive. 

Due to the high inoculum pressure, the lacks of certain fungicides and the massive 

dispersion there plants resistant against C. gloeosporioides have to be developed. To 

evaluate the resistance we present practicable and reproducible pathogenicity tests. 

P66 

Stephanie Nehrlich 

Sylvia Plaschil 

Reiner Krämer 



and Fruit Crops, Julius 

Kühn-Institute, Federal 

Research Centre for 

Cultivated Plants, Erwin- 

Baur-Strasse 27, D-06484 


stephanie.nehrlich@jki. 

bund.de 

Session Posters 139

P67 

Masahiro Nishihara 

Takashi Nakatsuka 

Yoshiko Abe 

Saburo Yamamura 

Cell engineering, Iwate 


Center, 22-174-4, Kitakami, 

Iwate, 024-0003, Japan 

mnishiha@ibrc.or.jp 

PRODUCTION OF BOUQUET-TYPE 

LISIANTHUS BY OVEREXPRESSION OF A 

RICE MADS BOX GENE OSMADS1 

Lisianthus is a very famous cut flower and many useful cultivars have been produced by 

conventional breeding. Here we attempted to produce new lisianthus with modified 

flowering traits by genetic engineering. Transgenic lisianthus (Eustoma grandiflorum 

Griseb. cv. Glory White) plants overexpressing a rice MADS box gene (OsMADS1) were 

produced by Agrobacterium-mediated transformation using bar gene as a selectable 

marker. The obtained plants showed remarkable shortened flower stalks and set bi or triflowers 

on one stem. Molecular analyses by Southern, northern and western blots 

confirmed that the foreign OsMADS1 gene was stably integrated into the lisianthus 

genome and expressed, resulting in the accumulation of the OsMADS1 protein in the 

transformants. Because some other morphological and physiological changes such as 

dwarf, reduction of flower size and change of flowering time were also observed in 

primary transformants, self-pollination was performed to evaluate the traits in the next 

generation. Two representative T 1 progeny lines were selected and subjected to the 

cultivation experiment under a closed greenhouse condition. The results demonstrated 

that the phenotype with short flower stalks was inherited in both transgenic lines. 

Furthermore, they had statistically different plant height, flower size and number of 

flowers compared with untransformed control plants. Flowering time was also changed in 

the two transformant lines. The obtained transformants are very unique and might be 

useful as bouquet for flower arrangement. These results demonstrated that genetic 

engineering could successfully introduce new traits to lisianthus as is the case with 

horticultural crops. 

140 Session Posters

POTENTIAL COMMERCIAL VALUE FROM 

MUTATED CLONES OF CALADIUM 

HUMBOLDTII SCHOTT ‘PHRAYA SAVET’ 

FROM IN VITRO CULTURE 

Mutated clones of Phraya Savet caladium (Caladium humboldtii cv. ‘Phraya Savet’) from 

in vitro culture were observed. Callus and small shoots were induced from unexpanded 

leaf segments cultured on modified Murashige and Skoog medium (MS) supplemented 

with 2.69 µM 1-Naphthalene acetic acid (NAA) and 17.76 µM N 6 -Benzyladenine (BA) for 

4 – 5 months with subculturing every 6 weeks. Shoots were transferred onto modified 

MS medium supplemented with 8.88 µM BA for shoot multiplication. Subsequently, roots 

were induced on MS without growth regulator for 2 weeks. The regenerated plantlets 

were vigorously grown in glasshouse conditions. From 3 morphological groups, leaf 

pattern, petiole and leaf colour were used to identify the mutated clones. The 

regenerated ‘Phraya Savet’ caladium plants were divided into 11 types. Variations were 

found in 10 types with multiple variants from 1 – 4 characters. The occurrence of variants 

was 34 percent. Leaf pattern variants were observed at the highest frequency of 28 

percent while leaf colour variants and petiole variants were found 16.0 and 4%, 

respectively. The most significance for commercial value from mutated clones was round 

leaf (9 percent). Other interesting variations were discussed. 

P68 

K. Obsuwan 

C. Thepsithar 


U. Somkanea 








Session Posters 141

P69 

Teresa Orlikowska 

Marta Zawadzka 

Research Institute of 


96-100 Skierniewice, 

Pomologiczna 18, Poland 

Teresa.Orlikowska@ 

insad.pl 

IN VITRO SELECTION OF ANTHURIUM 

ANDREANUM FOR SALT STRESS 

RESISTANCE 

Salinity is a troublesome factor for plants growing in soil-less substrates, such as 

anthurium grown in greenhouse conditions. To overcome this problem, it is necessary to 

grow genotypes that are more tolerant to salt stress. In vitro selection of some crops for 

tolerance of salt stress has proven successful, so we attempted to apply this technique 

to anthurium. The high potential of anthurium for adventitious regeneration also 

supported this approach. 

The youngest leaves, petioles and roots of micropropagated anthurium explants of 

the Bolero cultivar were in vitro incubated on media consisting of half-strengthMS 

(Murashige and Skoog, 1962) salts except ammonium nitrate lovered to ¼ concentration 

and different combinations of growth regulators. In one set of experiments, NaCl was 

included in the medium at concentrations of 0, 10, 20, 40, 80, 160 or 320 mM, and in the 

other at 0, 25, 50 or 100 mM 

The most effective medium for shoot regeneration was that containing 0.5 mg/l 

thidiazuron (TDZ) and 0.5 mg/l ά-naphthalene acetic acid (NAA). The earliest 

regeneration was from leaves, then from petioles and roots. The highest NaCl 

concentration in which survival and regeneration occurred was at 40 mM when BAP and 

2,4-D were used, and at 50 mM when TDZ and NAA were used. Shoot regeneration was 

usually preceded with globular callus, which was light green on the medium without 

NaCl, and light yellow with sporadic green points on the medium with NaCl. Green 

shoots on the media with NaCl regenerated 4-6 weeks later than those on NaCl-free 

medium. Their subculture on medium without growth regulators was performed for 

elongation, and with auxin for rooting, in both cases with the addition of the same 

concentration of NaCl on which the shoots had been induced. Most of these shoots 

yellowed and proved to be escapers, but some were still green after rooting. They will be 

tested for NaCl tolerance repeatedly before and after acclimatization in the greenhouse. 

142 Session Posters

ESTABLISHMENT OF PLANT REGENERATION 

SYSTEM AND AGROBACTERIUM-MEDIATED 

GENETIC TRANSFORMATION IN DAHLIA 

Dahlia (Dahlia hybrida) is a popular tuberous ornamental plant belonging to Asteraceae 

family. It has many cultivars with wide variations in flower colour, size and shape and 

grown for cut flower, and as pot and garden plants. For the cultivation, dahlias have 

many problems such as diseases and pests. Especially, viral diseases are the most 

serious since many cultivars are always facing to extinction because of the high 

susceptibility to some viruses such as dahlia mosaic virus. Although dahlia has high 

variations in flower colour and morphology, there have been constant demands for novel 

type of flowers such as blue flowers. To solve these problems, it is now expected to 

utilize genetic transformation methods. In this study, we aimed to develop an efficient 

protocol for in vitro plant regeneration and Agrobacterium-mediated genetic 

transformation. Calli were successfully induced from leaf on MS (Murashige and Skoog, 

1962) medium supplemented with 10 mg/l TDZ and 30 mg/l sucrose and shoot formation 

was observed after transferring the calli onto hormone-free medium. Based on these 

result calli induced on MS medium supplemented with 10 mg/l TDZ were inoculated with 

Agrobacterium tumefaciens strain EHA101 (pIG121-Hm) harboring both β-glucuronidase 

(GUS) and hygromycin resistant genes. After 2 days of co-cultivation, the calli were 

transferred to a selection medium containing hygromycin with meropenem for bacterial 

elimination. Survived calli were successfully regenerated on hormone-free medium 

without hygromycin. The regenerated shoots rooted on hormone-free medium containing 

hygromycin. The hygromycin-resistant plants thus obtained showed histochemical blue 

staining for GUS. Transformation of calli were confirmed by PCR and Southern blot 

analyses. 

P70 

Y. Otani 

D.P. Chin 

M. Mii 






271-8510, Japan 

yu_ukokshta@yahoo.co.jp 

Session Posters 143

P71 

Jae Ok. Park (1) 

Hye Sung Cho (1) 

Moon Young Park (1) 

Youn Seop Jo (1) 

Byung Joon Jeong (1) 

Geung Joo Lee (2) 


(1) Jeonnam Agricultural 

Research and Extension 

Services, Naju 520-830, 

Korea 

(2) Advanced Radiation 



Institute, Jeonbuk 580-185, 

Korea 

parkjo63@kornet.net 

CHARACTERISTIC OF AN ASTER 

SPATHULIFOLIUS MUTANT DERIVED FROM Γ- 

RAY TREATMENT 

Aster spathulifolius is a multi-annual growing evergreen herb and can be easily found in 

the dry soil condition of the coastal area of Korea and Japan. It generally has heavily 

dense pubescences on leaves and shoots, grows up to 50cm height and flowers violet 

from late September to early December. 

In an attempt to develop wild Aster spathulifolius into a pot plant with higher 

ornamental value and more suitable to flower bed of street, γ-ray was used for the 

mutation induction. 

The proper treatment dosage of γ-ray (LD 50 ) seemed as 30Gy, which have shown 

54% survival after the irradiation of γ-ray on the seeds. 

However, from the continuous observations on the whole plant population of M 1 V 1 

generation which have been irradiated in the process of dosage determination, we luckily 

selected one leaf mutant with golden coloured leaf margin among the 10Gy treated plant 

population. 

This selected mutant has not showed coherent leaf pattern till M 1 V 5 generation but 

the characteristics of this plant could be kept by only clonal propagation successfully. 

The seeds from the mutant were segregated phenotypically in the following generation. 

144 Session Posters

A DIGITAL IMAGE ANALYSIS SYSTEM (DIAS) 

FOR ASSESSMENT OF BIOASSAYS ON 

RHODODENDRON SIMSII AGAINST 

CYLINDROCLADIUM SCOPARIUM 

Cylindrocladium scoparium MORGAN belongs to the most important diseases of 

Rhododendron simsii. Successful controlling by breeding for resistance to this disease 

needs sensitive, practicable and reproducible screening methods. A research project 

aimed to develop effective screening methods for evaluation of plant resources for 

Cylindrocladium resistance in Rhododendron simsii will be presented. 

Bioassays with detached leaves were established. Drop inoculation was applied on 

the injured leaf base with C. scoparium suspension in a concentration of 10 6 spores per 

ml. The leaves were placed in Petri dishes on filter paper and the incubation was carried 

out at 22 °C over a period of 14 days in climate chambers. 

Development of disease symptoms could be quantified with a digital image analysis 

system (DIAS). All samples were photographed by a digital camera under defined 

illumination conditions 3, 7 and 14 days after inoculation. The software disperses and 

divides the image into colour grades according to the individual colour calibration. The 

calibration depends on the target of the bioassay and the variability of the symptom 

expression in the samples while users can define classes for their own discrimination 

needs. In some cases the differentiation between the healthy tissue and tissue infested 

by C. scoparium or the calculation of the disease severity ratio over the time series was 

highly problematic. As result of the analysis the proportional area of the respective colour 

grades were evaluated and transferred to a data base system. Afterwards the original 

data were transferred into the Excel programme and prepared for statistical analyses. 

This method allowed the accurate assessment of individual plant reaction to C. 

scoparium. 

87 Rhododendron genotypes (65 R. simsii varieties and 25 Rhododendron species) 

were screened in the bioassays to C. scoparium respectively. The responses of the 

genotypes to C. scoparium were estimated by symptom scoring with the digital image 

analysis system (DIAS). The analyses of the disease symptoms showed significant 

differences regarding the susceptibility and disease progress to the fungal pathogen. 

P72 

S. Plaschil 

R. Krämer 



and Fruit Crops, Julius 

Kühn-Institute - Federal 

Research Centre for 

Cultivated Plants, Erwin- 

Baur-Strasse 27, D-06484 


Sylvia.plaschil@jki.bund.de 

Session Posters 145

P73 

Juozas Proscevičius (1) 

Violeta Kleizaite (2) 

(1) Department of Natural 

Science, Vilnius 

Pedagogical University, 

Studentu 39, 08106 Vilnius 

Lithuania, Institute of 

Botany, Zaliuju Ezeru 49, 

08406 Vilnius, Lithuania 

(2) Department of Natural 

Science, Vilnius University, 

M. K. Ciurlionio 21/27, 2034 


juozas.proscevicius@ 

gmail.com 

INVOLVEMENT OF SPECIES LILIUM 

CANDIDUM IN BREEDING OF LILIES 

L. candidum is cultivated as ornamental plant from ancient time. However, despite high 

ornamental value and potency of vegetative propagation this species is limited by 

demand to environmental conditions and by sensitivity to fungal and virus infections. 

Reproduction isolation barriers are the main reason why L. candidum is rare involved for 

breeding of lilies. In this investigation L. candidum was involved in incongruous crosses 

with Asiatic Hybrids (AH), Trumpet Hybrids (TH) and L. longiflorum. The cut style 

pollination and pollination by mixed incongruous pollen were used as methods to 

overcome pre-fertilization barriers. Cultivation of isolated embryos in vitro allowed rescue 

hybrid progeny. 

Neither native nor cut style pollination allow achieve fertilization in crosses L. 

candidum x AH or AH x L. candidum. However when pollen of L. candidum was used in 

mixtures with pollen of other incongruous species to perform pollination of AH female 

fertilization was successful. The progeny derived from such crosses were screened by 

inheritance of isozymes patterns and DNA markers. Several tens plants were selected 

as AH x L. candidum hybrids by superoxide dismutase (SOD) patterns inherited from L. 

candidum. Some progeny were received as descendant after pollination of TH with 

mixed pollen of L. candidum and L. monadelphum and several plants were received by 

cut style pollination of L. longiflorum with L. candidum. However it‘s hybrid nature must 

be confirmed. 

Morphology and grow vigour of young hybrid plants AH x L. candidum grown in soil 

condition was in common with AH. They don’t form basal leave rosette which is 

characteristic to paternal species L. candidum. The flowers of solitary flowered young 

hybrid plants were something intermediate in form between maternal AH and parental L. 

candidum. Some of AH x L. candidum hybrids were converted in allotetraploid forms to 

involve them for backcrosses with AH and L. candidum. 

146 Session Posters

SOMACLONAL VARIATION IN 

MICROPROPAGATED TULIPS AS A SOURCE 

OF NOVEL GENOTYPES – FIELD AND 

MOLECULAR CHARACTERISTIC 

The aim of the study was to evaluate somaclonal variation (SV) in tissue-culture (TC) 

derived plants of three tulip genotypes: ‘Giewont’, ‘Prominence’ and a mutant ‘Bs6’. The 

mutant ‘Bs6’ was selected from among the micropropagated plants of the cultivar ‘Blue 

Parrot’ in 2004. Compared to ‘Blue Parrot’ true-to-types the plants of this new genotype 

have flowers longer by 1.5-2 cm, stems longer by 5 cm and colour of flowers changed 

from purple-violet to red-violet. The plant material of all genotypes derived from the longterm 

cultures maintained in vitro for the period of 2-4 years for ‘Giewont’ and 

‘Prominence’, and 6 years for ‘Bs6’. The TC-derived plants were planted outdoors in an 

insect-proof tunnel in 1999-2004. During the subsequent years of cultivation 2003-2008 

phenotypic evaluations were done when the plants were in full bloom. Juvenile plants 

were examined for leaf abnormalities. The reference plants for ‘Giewont’ and 

‘Prominence’ were propagated conventionally while for ‘Bs6’ the reference plants were 

derived from in vitro culture. 

SV frequency depended on genotype and time of culturing in vitro. In case of 

‘Prominence’ the lowest SV frequency (5.3-9.2%) was observed for plants derived from 

the two-year cultures and the highest one (28.2-48.9%) for plants from four-year 

cultures. In case of ‘Giewont’ and ‘Bs6’ the SV frequency ranged from 6.7% to 13.8%. 

Most of the off-type plants out of all genotypes had minor changes. The colour of flowers 

was unchanged, however, the shape of flowers was slightly altered, e.g. in some 

‘Prominence’ variants, tepals had acute tips or in ‘Bs6’ variants, a tepal goffering was 

atypical. 

In all the studied genotypes, phenotypic evaluation showed a regular occurrence of 

variants with major changes such as highly malformed flowers. The colour of flower of 

these variants was unchanged, while tepals were irregularly notched and had white 

stripes. All the variants had leaves with thicken, vitreous venation. Such leaves were 

also found in some juvenile plants of all the genotypes. DNA analysis with an use of 

inter-simple sequence repeats (ISSR) carried out on the somaclonal variants with major 

changes of both juvenile and flowering plants confirmed that changes in leaf morphology 

resulted from genetic changes. The obtained results indicate that the trait of the leaf 

thicken, vitreous venation can be considered as the morphological marker for early 

detection of the major genetic changes within juvenile plant material. 

P74 

Małgorzata 

Podwyszyńska 

Anita Kuras 

Krzysztof Niedoba 

Małgorzata Korbin 

Research Institute of 


Pomologiczna 18, 96-100 

Skierniewice, Poland 

mpodwysz@insad.pl 

Session Posters 147

P75 

Nancy Pyck 

Johan Van Huylenbroeck 

Institute for Agricultural and 

Fisheries Research (ILVO) – 

Plant Unit Sciences Unit, 

Applied Genetics and 



johan.vanhuylenbroeck@ilvo. 

vlaanderen.be 

BUSINESS 2 RESEARCH: AN EFFICIENT 

SYNERGY 

Ornamental production is a very innovative sector. A prominent bottleneck however is 

the transfer and implementation of expertise present in research institutes to the 

hortibreeder business. On the other hand, introducing new cultivars in a successful 

manner is complex for public breeding institutes. In order to overcome this hurdle, a 

diverse range of long-term partnerships between ornamental entrepreneurs and 

research institutes was set up. 

These close interactions can make the innovation process of companies feasible 

and more straight forward. Growers that take part in such collaborations between 

business and research pay therefore a yearly contribution, which is in turn directly 

invested in breeding activities and research. In this context, a successful project 

offering technological advice and service in horticulture, called SIETINET, has been 

running for already 5 years. Likewise, AZANOVA cvba and BEST-select cvba, involving 

innovative breeding in respectively pot azalea and woody ornamentals, are great 

examples of how research institutes can play a vital role in empowering a continuous 

strategy of innovation within hortibreeder business. 

Offering expert technological advice constitutes a central component within 

SIETINET. Another interesting service of the network is the possibility to design testing 

services, defined by the grower, that require specialized scientific equipment. In the 

past, successful experiments have been set up for e.g. the fine tuning of the growing 

systems for new cultivars, the application of methods for ploidy alteration, mutation 

breeding, in vitro techniques to overcome interspecific crossing incongruities, study on 

rooting and acclimatization of candivars. In breeding activities of AZANOVA and BESTselect, 

the participating growers are involved at an early stage in evaluating new 

candidate cultivars. Equally important for these partnerships is the subsequent lucrative 

commercialization of the resulting product innovations. Consequently, efforts are made 

to attract media coverage and to improve access to markets. 

As technological innovation and improvement play a crucial role in hortibreeder 

companies, services offered by these partnerships may have an important impact on 

members R&D investments and makes it possible for growers to get access to new 

cultivars. 

148 Session Posters

MARKER ASSISTED BREEDING FOR 

NEMATODE RESISTANCE IN HYPERICUM 

Root-knot nematode (Meloidogyne spp) is an important plant parasite in Hypericum L. 

So far, there are no acceptable cut flower or pot varieties with nematode resistance. 

Given that recently demand for Hypericum has risen mainly due to its versatility as a filler 

in mixed bouquets, there is an increased need for developing nematode resistant 

varieties. Introduction of nematode resistance will enable the production of varieties with 

improved performance and quality, without the use of dangerous nematicides and soil 

disinfection methods that are harmful to people and the environment. The aim of this 

project is to introduce nematode resistance in cultivated Hypericum, to identify molecular 

markers associated with nematode resistance and develop marker assisted selection in 

this crop. 

Field assays with soil infested with root-knot nematode and evaluation of disease 

and pathogen levels have led to the identification of six resistance species. Out of these 

two resistant genotypes have been chosen for a crossing program. Four interspecific 

reciprocal crossings are in progress, between two cut flower varieties and the two 

nematode resistant genotypes. GISH will be used for verification of hybrid character of 

the crosses and assessment of the number of introduced chromosomes from the wild 

species and possible recombination between the species. For phenotyping, 100 F1 

genotypes per crossing will be evaluated in an experimental field that will be uniformly 

infested by nematodes. For this purpose, a total of three replicates of 25 plants per 

genotype will be sown and the plants will be grown for two consecutive flowering 

periods. For genotyping, identification of associated molecular markers of nematode 

resistance will be performed with NBS-profiling, a molecular marker technique that 

efficiently produces markers in resistance genes by targeting towards conserved regions 

of these resistance genes. 

P76 

Ana M. Quiñones (2) 

María E. Guerrero (2) 

Fernando Ponce (2) 

Pedro Kingman (2) 






International, Wageningen, 


(2) Esmeralda Breeding 

BV, The Netherlands and 

Ecuador, Río Amazonas 

7714 y Río Curaray, Quito, 

Ecuador 







anamq@hilsea.com.ec 

Session Posters 149

P77 

Varahram Rashidi (1) 

Mahyar Yousefzadeh (2) 

Ahmad Razban 

Haghighi (3) 

Hamideh Javadi (3) 


Agronomy and Plant 

Breeding, Faculty of 

Agriculture, Islamic Azad 

University, Tabriz Branch, 

Iran 

(2) M.Sc. Graduated of 

Plant Breeding, Islamic 

Azad University, Tabriz 

Branch, Iran 

(3) Natural Resources 

Research center, Tabriz, 

Iran 

rash270@yahoo.com 

CYTOGENETICAL STUDY OF CHAMAN 

SHOUR SAHELI (AELUROPUS LITTORALIS) 

The Chaman Shour is plant that has growing in around of Erumeih pond, Iran. Whereas 

it's one of halophyte plant and also can use thereof for nutrition in ranch, therefore is 

important. In order to accustom with chromosome characteristic's its karyological studied 

in Islamic Azad University- Tabriz Branch in 2006. In this study have used from root apex 

meristematic cells. The results showed that basic number of chromosomal in this plant 

is fifteen (x = 15), kind of chromosomes are metacentric and its ploidy is diploid. The 

long of tallest Chromosome (number 1) is 3.0354 + - 0.12 micron and long of shortest 

Chromosome (number 15) have estimated 1.3684 + - 0.04 micron. The total long of 

genome have estimated 29.8619 micron. 

150 Session Posters

ACQUISITION OF OTO, OA AND FO 

INTERSPECIFIC HYBRID LILIES AS A 

SUBSTITUTE FOR ORIENTAL HYBRIDS BY 

OVULE CULTURE 

Oriental hybrid lilies are popular cultivated in Korea and exported to Japan’s flower 

market. However, it is very weak growth hot summer season, weak disease (ex. Virus 

and Fusarium) resistance, long breeding and bulb growth period and strong fragrance in 

Korea. To introduce new genetic variation and substitute for disadvantage of Oriental 

hybrids, it has crossed with Lilium OT interspecific hybrids; L. x formolongi hybrids and 

Asiatic hybrids by cut-style and stigmatic pollination method and ovule culture since 

1997. For the first time, four new OTO (Lilium OT interspecific hybrid x L. Oriental 

hybrids) hybrids were bloomed in 2008. The parents of ‘OTO-08-2’ and ‘OTO-08-2-2’ are 

between ‘Avocado’ and ‘Medusa’. The characteristics of ‘OTO-08-2’ have oriental flower 

shape, male sterile, strong growth and weak fragrance. Flowers are upward-facing and 

thicken red pink and ivory edge (RHS, 67P, W155B). The characteristics of ‘OTO-08-2-2’ 

have oriental flower shape, male sterile, and fragrance. Flowers are sideward-facing and 

reddish pink and ivory edge (RHS, R-51C, and W-155A). The parents of ‘OTO-08-1’ are 

between ‘Avocado’ and ‘Cordoba’. The characteristics of ‘OTO-08-1’ have oriental flower 

shape, weak fragrance and male sterile. Flowers are upward-facing and white with ivory 

centered (RHS, W155B). The parents of ‘OTO-08-3’ are between ‘Avocado’ and 

‘Acapulco’. The characteristics of ‘OTO-08-3’ have oriental flower shape, male sterile, no 

pollen grains, and weak fragrance. Flowers are upward-facing and red with light salmon 

(RHS, R-46A, Y11G, and D). OA-05-1 was obtained between Oriental hybrid ‘Casa 

Blanca’ and Asiatic hybrid ‘Sgl pepper’. Characteristics of OA-05-1 were upward-facing 

flowers, weak fragrance, purple flower colour (RHS, RP60B). FO progenies between 

‘Raizan’ and ‘Oriental O-54 line’ are obtained FO-00-1, FO-00-3, FO-00-4, FO-00-6, FO- 

00-10, FO-00-12 and FO-00-16. Phenotypic characteristics of FO progenies were similar 

to that of Oriental hybrids such as diverse colour distribution from creamy ivory as 

female flower colour to deep pink as male flower colour, Oriental flower shaped and leaf 

shaped, flower size, weak fragrance and so forth. Among new diverse interspecific 

hybrids has been obtained, OTO hybrids will be adequate cultivars adapted to the 

Korean climate and environmental conditions. 

P78 

Hye Kyung Rhee 

Hae Ryong Cho 


Horticultural and Herbal 

Science, RDA; (#441-440) 

540-41, Top-dong, 

Kwonsun-Gu, Suwon, 

Republic of Korea 

rheehk@rda.go.kr 

Session Posters 151

P79 

Christina Rode (1) 

Traud Winkelmann (2) 

Hans-Peter Braun (1) 

(1) Institute of Plant 

Genetics, Leibniz University 

Hannover, Herrenhaeuser 

Str. 2, D-30419 Hannover, 

Germany 

(2) Institute of Floriculture 

and Woody Plant Science, 

Leibniz University 

Hannover, Herrenhaeuser 

Str. 2, D-30419 Hannover, 

Germany 

rode@genetik.unihannover.de 

ESTABLISHMENT OF PROTEOME 

REFERENCE MAPS FOR SOMATIC AND 

ZYGOTIC EMBRYOS OF CYCLAMEN 

PERSICUM MILL 

Cyclamen persicum is a popular ornamental crop with a high economic relevance for the 

horticulture business. Within Cyclamen persicum somatic embryogenesis has been 

shown to be an efficient vegetative propagation system and the development of artificial 

seeds is an ultimate aim. For this approach, somatic embryos have to be produced 

which mimic their zygotic counterparts in protein composition. Moreover, the physiology 

of somatic and zygotic embryogenesis remains unclear in Cyclamen up to now. Initial 

analyses comparing the proteomes of somatic and zygotic embryos have been 

performed previously. Approximately 70 % of the proteins expressed in zygotic embryos 

also were present in somatic embryos in comparable abundance. ESI-MS/MS analysis 

led to identification of 20 proteins of both tissues. 

Based on these analyses, further comprehensive proteomic characterizations were 

performed aiming to create proteome reference maps for somatic and zygotic embryos 

of Cyclamen persicum. Separation by two dimensional IEF-SDS gel electrophoresis 

leads to a resolution of more than 700 protein spots for each tissue. Consistent to the 

initial analysis, approximately 

70 % of the spots likewise appeared in both zygotic and in somatic protein fractions. 

However, DIGE analysis revealed extremely high alterations in abundances for the 

majority of proteins present in both tissues. MS analysis for total 300 reproducible spots 

(260 of the zygotic embryos' protein fraction and additionally 40 spots appearing 

specifically in the somatic embryos' proteome) led to identify 261 proteins, whereof 33 

specific for the somatic tissue. 

Currently, identified proteins are clustered according to their physiological relevance 

and a digital proteome reference map is created. Additionally, shotgun proteomics are in 

preparation for both embryo tissues. Also proteomic analysis concerning the 

development and maturation of somatic embryos are in progress. 

152 Session Posters

MUTATION FREQUENCY INDUCED BY 

CHEMICAL MUTAGENS IN M 3 GENERATION 

OF PETUNIA X ATHINSANA D.DON 

The seeds of cultivar (‘Flash Red’) of Petunia x athinsuna D. Don were used as the 

initial material for the studies. Mutations were induced by soaking these seeds for 1 hour 

in the solution with pH 4 of EMS (concentration 0.5 and 1.5 mM), MMS (1.5 and 2 mM), 

DES (0.5 and 1 mM) and AS (1 - 1.5 mM) at the presence of buffer orthophosphoric acid 

at the concentration 0.025 mM. In order to check the genetic background of the 

observed mutations in M 3 generation DNA samples were taken for analysis from the 

plants with the phenotype different from control plants. Variability on the DNA level was 

determined by ISSR-PCR technique. 

In M 3 generation of mutants the most frequently occurring changes were white tiny 

spots or lines on corolla petals, flower colour change from red into pink, chlorophyll leaf 

changes and irregular corolla rim. The majority of M 3 phenotypic variation had already 

been found in M 2 generation. Chemical mutagens had a stimulating effect on the number 

of flower buds. The greatest number of flower buds in mutants resulted from the 

application of 0.5 mM of EMS and 0.5 mM of DES (520 – 530% of control) for 

mutagenesis. 

In most cases phenotypic changes in M 3 generation were of genetic nature. Their 

genotype similarity ranged from 4.9- 25.5 %. Most changes of this kind were obtained 

after 0.5 mM EMS and 0.5 DES had been used for inducing mutation. 

P80 

D. Rzepka-Plevneš 

M. Krupa-Małkiewicz 

K. Zając 

Agricultural University, ul. 

Janosika 8, 71-424 

Szczecin, Poland 

rzepka@agro.ar.szc 

zecin.pl 

Session Posters 153

P81 




Tomoya Niki (1) 


Takuma Komatsu (1) 

Takaaki Nishijima (1) 


(2) 




National Agriculture and 

Food Research 

Organization, Tsukuba, 

Ibaraki 305-8519, Japan 



Advanced Industrial 



Japan 

kattu@affrc.go.jp 

FUNCTIONAL ANALYSIS OF TORENIA CLASS 

B GENES, TFGLO AND TFDEF: THEIR UNIQUE 

TRANSGENIC PHENOTYPES AND TARGET 

GENE REGULATION 

We are studying functions of floral organ identity genes using torenia (Torenia fournieri 

Lind. ‘Crown Violet’), to establish efficient procedure for controlling floral traits by genetic 

engineering. In this study, we focused on TfGLO and TfDEF, the torenia class B genes, 

those have been supposed to play important roles in formations of petals and stamens. 

To understand the roles of the two genes, we firstly generated a set of class B geneoverexpressing 

transgenic lines and -repressed lines. The latters were generated by 

introducing chimeric repressors with the EAR-motif repression domain (SRDX; Hiratsu et 

al. 2003). TfGLO-overexpressing lines showed the accumulation of anthocyanin 

pigments in sepals, and TfGLO-repressed lines exhibited distinctive serration in petal 

margins. In TfDEF-repressed lines, irregular pigmentation pattern along the vascular 

bundles was observed within the purple segment of petals, and no phenotypic change 

was observed in TfDEF-overexpressing lines. 

We next examined the expressions of putative target genes of TfGLO and TfDEF, in 

accordance with previous Arabidopsis study. We isolated xyloglucan endo-1,4-beta-Dglucanase 

genes (TfXEG1, TfXEG2 and TfXEG3) and chlorophyll a/b binding protein 

genes (TfCAB1, TfCAB2 and TfCAB3) from torenia, then performed RT-PCR analysis 

using floral organs of the transgenic lines. As we have expected, the expressions of 

TfXEG1, 2 and 3 were up-regulated in class B gene-overexpressing lines and downregulated 

in class B gene-repressed lines. On the contrary, the expressions of TfCAB1, 

2 and 3 were not influenced by overexpressing the class B genes, while their 

expressions were up-regulated in class B gene-repressed lines. 

These results suggest that both TfGLO and TfDEF participate in the developmental 

process of floral organs, while they seemed to have different functions. In addition, 

putative target genes of class B genes, such as TfXEGs, were up-regulated in torenia as 

same as in Arabidopsis. Now, we are focusing on the regulation of anthocyanin 

biosynthesis by class B genes because anthocyanin pigmentation in sepals was 

observed only in TfGLO-overexpressing lines. To understand further the individual 

functions of the two class B genes, we intend to examine the expressions of anthocyanin 

biosynthesis-related genes using the class B genes-overexpressing and -repressed 

lines. 

154 Session Posters

INDUCTION OF 2N GAMETES AND 4N 

EMBRYO IN LILIUM (LILIUM × FORMOLONGO 

HORT. BY NITROUS OXIDE GAS TREATMENT 

The effect of nitrous oxide gas (N 2 O) treatment on the induction of 2n gametes and 4n 

embryo in Lilium × formolongo hort. was examined. Lilium × formolongo hort. is an 

artificial hybrid between Lilium longiflorum Thunb. and Lilium formosanum Wall. In 

general the members in the genus Lilium is propagated vegetatively. However, Lilium × 

formolongo hort. is exceptionally propagated by seeds and blooms 6 - 8 months after 

sowing. Seeds of Lilium × formolongo hort. cultivars ‘Raizan’ and ‘Kitazawa-Wase’ 

were sown in a plastic tray with 200 holes on 5 February 2008 and seedlings bearing 2-3 

leaves were transplanted in 10.5cm (in diameter) pots on 5 May, grown under ambient 

conditions in a green house. When these first flower buds of ‘Raizan’ plants were 

reached at 15 – 30 mm in length, these buds were sampled consecutively and observed 

by the aceto-carmine squash method to determine the meiotic stage of pollen mother 

cells. The buds ranging from 19 – 23 mm in length contained pollen grains at prophase I 

to tetrad. Immediately after the staging of pollen grain, plants with the buds in the same 

length were treated with N 2 O gas for 48 h at 6 atm at room temperature. The diameter of 

mature pollen grains in metaphase, obtained from the plants with buds of 22mm in 

length at the time of exposure to N 2 O gas, ranged from 76 to 169, although those from 

untreated ones ranged from 62 to 95μm. For the induction of 4n embyo, thirteen days 

after the pollination with untreated pollen grains of ‘Kitazawa-Wase’, ovaries of ‘Raizan’ 

were treated with N 2 O gas for 72h at 6 atm. Plants with pollinated ovaries grew normally 

after the treatment with N 2 O gas and these capsules were harvested 75 - 85 days after 

pollination. We obtained 25 seedlings from these capsules and ramdomly selected five 

seedlings. Of five seedlings, two were revealed to be tetraploid by flow cytometry. We 

could demonstrate that N 2 O gas treatment was useful for the manipulation of both male 

gamete as well as zygote for the polyploidy breeding in Lilium × formolongo hort. 

P82 

T. Sato (1) 

K. Miyoshi (2) 

K. Okazaki (3) 

(1) Akita Pref. Agric. Exp. 

Stn., Yuwa Akita City Akita 

Pref. 010-1231, Japan 

(2) Faculty of Bioresource 

Sci., Akita Prefectural 

Univ., Akita City, Akita Pref. 

010-0195 


Niigata Univ., Niigata City, 

Niigata,Pref. 950-2181, 

Japan 

satou-takao@pref. 

akita.lg.jp 

Session Posters 155

P83 


Takako Narumi (2) 






(3) 





Food Research 

Organization, Tsukuba, 

Ibaraki 305-8519, Japan 


Kagawa University, Mikicho, 

Kagawa 761-0795, 

Japan 



Advanced Industrial 



Japan 

mshikata@affrc.go.jp 

EFFICIENT SCREENING OF TRANSGENIC 

TORENIA WITH NOVEL FLORAL TRAITS BY 

COLLECTIVE INTRODUCTION OF CHIMERIC 

REPRESSORS FOR ARABIDOPSIS 

TRANSCRIPTION FACTORS 

Molecular breeding enables us to produce novel floral traits of horticultural plants which 

could not be obtained by traditional breeding. Since a number of transcription factors, 

such as MADS-box and TCP family proteins, play a role in the control of flower 

development, transcription factors are useful tools for the manipulation of floral traits and 

generation of novel variation. Although we usually need sequence information of the 

gene that we try to use, horticultural plants have little information of genome and EST. 

By contrast, information of Arabidopsis, the model higher plant, is abundant and 

facilitates our molecular breeding. Accumulating facts that chimeric repressors of 

Arabidopsis transcription factors altered floral traits of some horticultural plant species 

may lead to an innovation of molecular breeding. On the other hand, it is necessary to 

develop an efficient screening method because we cannot find out useful genes for 

valuable floral traits without regenerating transgenic plants. To obtain new floral traits 

efficiently, we selected 42 transcription factor genes which highly expressed in 

Arabidopsis flowers and collectively introduced their chimeric repressors into torenia 

(Torenia Fournieri Lind. cultivar ‘Crown Violet’). We screened phenotypically-altered 193 

lines of 348 transgenic plants. We found that 82.4% of them had single transgene, and 

39 of 42 constructs were introduced independently. One third of transgenic plants with 

single transgene induced recognizable phenotypes in petal colour and/or shape as 

expected, such as white margined petals, uniformed colour corolla, and partly opened 

flower. These results indicate that bulk collective introduction of chimeric repressors of 

Arabidopsis transcription factors makes possible the efficient production of novel 

horticultural plants with valuable traits. Some transcription factors used in this study 

seem to act in the same regulatory pathway because similar phenotypes were observed 

in some transgenic plants harboring different chimeric repressors. Most of selected 42 

genes were functionally unknown, but phenotypes in transgenic torenias would provide 

information which cannot be revealed by analysis in Arabidopsis. Now we are 

transforming another set of 50 genes selected with different approach to explore further 

variation of floral traits. 

156 Session Posters

ACCUMULATION, PRESERVATION AND 

INVESTIGATION OF LITHUANIAN 

ORNAMENTAL PLANTS IN THE BOTANICAL 

GARDEN OF VILNIUS UNIVERSITY 

Accumulation, preservation and investigation as well as conservation of genetic 

resources have become a prestigious task for all the countries of the world. By signing 

the Rio Convention on Biological Diversity on June 11, 1992, Lithuania has committed to 

conserve its genetic resources. In 1993 National Coordination Centre for Conservation 

of Plant Genetic Resources at the Lithuanian Agriculture Institute was established. 

Researches of the Floriculture Department of the Botanical Garden of Vilnius University 

joined the program of investigation and conservation of genetic resources in 1994. At 

present the gene fund collection consists of 900 flower taxa developed by Lithuanian 

plant breeders. The authors of Lithuanian flower cultivars (both amateurs and 

professionals) have created priceless national wealth i. e. flower cultivars and hybrids. 

Therefore, collection, preservation, investigation and evaluation of Lithuanian flower 

genefund is a new trend of scientific researches not only in the botanical gardens but 

also in the whole country. 

Lithuanian flower breeders released a lot of new cultivars of Crocus L., Dahlia Cav., 

Gladiolus L., Hemerocallis L., Iris L., Lilium L., Paeonia L., Primula L., Tulipa L. and etc. 

The aim of this research is to study and evaluate ornamental properties of flower 

cultivars released by the Lithuanian breeders. 

The investigations, descriptions and evaluations of morphological, bioecological and 

ornamental properties of gladiolus (Gladiolus L.), lily (Lilium L.), iris (Iris L.), primrose 

(Primula L.), dahlia (Dahlia Cav.), peony (Paeonia L.) cultivars were carried out in the 

period of 1998 – 2008 according to the requirements of the International Union for the 

Protection of New Varieties of Plants (UPOV) and methodologies used in neighbouring 

countries. The colour of the blossoms and leafs are determined according to the 

international R.H.S. (The Royal Horticulture Society) colour chart. 

Having collected, accumulated, researched and evaluated the flowers created by 

Lithuanian breeders, in the future it will be possible to select the most valuable cultivars 

(genetic resources) and to make a system of the effective preservation and rational 

usage of the genetic resources. In general, Lithuanian cultivars are originals adapted to 

the local climate conditions, and it is urgent to conserve, investigate and foster it as a 

part of the land culture. The most distinguishing and attractive (unbelievable form, 

perfect display of blooms in the spike, colour harmony) Lithuanian gladioli cultivars, 

researched in the Botanical Garden of Vilnius University, are the following ones: ‘Fiji’ 

(author A. Lukosevicius), ‘Nu, Gromov, Pogodi!’ (P. Ciplijauskas), ‘Laimute’ (P. 

Ciplijauskas), ‘Merkurijus’ (A. Lukosevicius), ‘Norma’ (A. Lukosevicius), ‘Onute-3’ (P. 

Balcikonis), ‘Paparcio Ziedas’ (P. Ciplijauskas), ‘Saules Takas’ (P. Balcikonis), 

‘Snieguole’ (P. Ciplijauskas), ‘Solveiga’ (A. Lukosevičius), ‘Spalvingas Sapnas’ (J.A. 

Liutkevicius). The results of Paeonia genus researches were summarized and 25 

hybrids originated by E. and J. Tarvidas were suggested to the register of national gene 

fund. 

P84 

G. Stukeniene 

R. Juodkaite 

S. Dapkuniene 

A. Skridaila 

Botanical Garden of Vilnius 

University, Kairenu 43, 

Vilnius, LT–2040, Lithuania 

Gitana.Indrisiunaite@gf.vu.lt 

Session Posters 157

P85 

Katsuhiko Sumitomo 

Satoshi Yoshioka 

Yu-ichi Fujita 

Atsuko Yamagata 

Takashi Onozaki 

Akemi Ohmiya 

Michio Shibata 




Food Research 

Organization, Fujimoto, 


Japan 

ksumi87@affrc.go.jp 

EXISTENCE AND FUNCTION OF PETAL 

COLOUR IDENTITY GENE IN APETALOUS 

WILD CHRYSANTHEMUM 

Chrysanthemum (Chrysanthemum morifolium) is one of the most important ornamental 

crops worldwide. In most chrysanthemums, the capitulum is composed of ray florets 

(petals) and disk florets. The ray floret colour in yellow-flowered cultivars is mainly due to 

carotenoids. Recently, Ohmiya et al. (2006) showed that the ray floret colour in whiteflowered 

cultivars is due to the degradation of carotenoids by the expression of 

CmCCD4a, a gene that encodes carotenoid cleavage dioxygenase; CmCCD4a 

suppression by RNA interference (RNAi) give yellow colour to white ray florets. 

In wild chrysanthemums, the ray florets are either white or yellow, indicating that ray 

floret colour probably results from crotenoid degradation in wild chrysanthemums, as in 

chrysanthemum cultivars. Southern blot analysis confirmed that wild chrysanthemum 

species with white ray florets had CmCCD4a homologues, while those with yellow ray 

florets did not. This suggests that carotenoid degradation also contributes toward 

producing ray floret colour in wild chrysanthemums. 

CmCCD4a expression was observed to be strictly limited to ray florets. Southern blot 

analysis showed that C. shiwogiku has a CmCCD4a homologue, although its capitulum 

has only disk florets and no ray florets. To confirm the function of this CmCCD4a 

homologue, C. shiwogiku was interspecifically hybridized with a cultivar, ‘Squash’, which 

does not have CmCCD4a and produces yellow ray florets. The capitulum of all progeny 

individuals produced white ray florets. CmCCD4a homologue fragments were detected 

in all progeny individuals by polymerase chain reaction (PCR). These results showed 

that the CmCCD4a homologue present in C. shiwogiku performed an enzymatic function 

in the ray florets. Further, these findings suggest that C. shiwogiku may have originated 

from a species with white ray florets by loss of genes related to ray floret formation. 

Similar to C. pacificum, C. shiwogiku produces numerous small capitula in each leaf 

axil, and this is a useful characteristic for breeding (De Jong, 1989). C. shiwogiku and C. 

pacificum have no ray florets, and therefore, breeders do not know the colour of ray 

florets in these species. Investigation of CmCCD4a homologue before crossbreeding 

can predict the ray floret colour of their progeny. 

158 Session Posters

MICROPROPAGATION OF SELECTED 

ORNAMENTAL PLANTS 

In the present work, three (3) popular ornamental plants in Malaysia were selected for 

micropropagation studies using tissue culture system. The species utilized were Begonia 

heimalis Fotsch, Gerbera jamesonii and Polianthes tuberosa. The scope of the study 

covers investigation on the role of various plant hormones such as Naphthalene acetic 

acid ( NAA), Indole acetic acid (IAA), kinetin, Benzyladenine (BA) etc on the in vitro 

morphogenesis of these species. The main aim being to regenerate these ornamental 

plants in vitro for mass propagation. Based on the results obtained, these species were 

very responsive in culture, they could form multiple shoots and roots quite readily and 

some could even produce in vitro flowering. The efficient regeneration systems were 

established for the three species on MS ( Murashige and Skoog, 1962) medium 

supplemented with various hormones at optimum concentrations. 

P86 


Nor Azlina Hasbullah 

Sakinah Abdullah 

Asmah Awal 



Science, University of 

Malaya, 50603 Kuala 

Lumpur, MALAYSIA 

rosna@um.edu.my 

Session Posters 159

P87 

Takejiro Takamura 

Natsuki Yoshimura 

Mayo Horikawa 

Faculty of Agriculture, 

Kagawa University, 2393 

Ikenobe, Miki-cho, Kagawa 

761-0795, JAPAN 

take@ag.kagawa-u.ac.jp 

PLOIDY LEVELS OF DEGENERATED 

EMBRYOS IN THE CROSSES BETWEEN 

DIPLOID AND TETRAPLOID CYCLAMEN 

The formation of unexpected tetraploid progenies and inhibition of triploid-seed formation 

in the reciprocal crosses between diploid and tetraploid cyclamen was reported. In the 

crosses, it was also been suggested that the tetraploid progenies were formed by the 

fertilization between a reduced gamete from a tetraploid plant and an unreduced gamete 

from a diploid plant, and that the development of many zygotic embryos were inhibited. 

Ploidy levels of the degenerated embryos were, however, obscure. Therefore, embryo 

rescue by the ovule culture in the crosses were examined. 

A lot of progenies were obtained in the 2x x 4x crosses, when the ovule culture by 

using medium with coconut water was done 28 days after pollination. Although almost all 

fruits were dropped 28 days after pollination in the 4x x 2x crosses, some progenies 

were obtained by the ovule culture at seven days after pollination. 

Almost all progenies obtained by the ovule culture after the reciprocal crosses 

between diploids and tetraploids were triploids, suggesting that many triploid zygotes 

were formed in the crosses. Some tetraploid, pentaploid and hexaploid progenies were 

also obtained in the 2x x 4x crosses after the ovule culture. Origin of the pentaploids was 

suggested the fertilization between a reduced gamete from a diploid plant and an 

unreduced gamete from a tetraploid plant. 

These results suggest that many triploid zygotes are formed in the crosses between 

diploid and tetraploid cyclamen, whereas development of the triploid embryo is inhibited 

without the embryo rescue. It is also indicated the possibility that pentaploids and 

hexaploid progenies as well as triploids and tetraploids are obtained in the crosses by 

using the ovule culture, depending on the cross combination. It should be suggested that 

development of the pentaploid and hexaploid embryos might be inhibited without the 

embryo rescue. 

160 Session Posters

INTERGENERIC HYBRIDIZATION AND 

RELATIONSHIP OF GENERA WITHIN THE 

TRIBE ANTHEMIDEAE CASS. (I. 

DENDRANTHEMA CRASSUM (KITAM.) 

KITAM.×CROSSOSTEPHIUM CHINENSE (L.) 

MAKINO) 

An intergeneric hybridization has been made between Dendranthema crassum (kitam.) 

kitam. (2n=90; ♀) and Crossostephium chinense (L.) Makino (2n=18; ♂). Most of the 

hybrid embryos aborted at an early developmental stage. Using ovule rescue, totally 160 

plump ovules (at 15d post pollination) were selected for in vitro culture, it was possible to 

establish a single intergeneric hybrid plant showing 2n=54 chromosomes. The leaf 

length, leaf width and epidermal hair density of the hybrid were all intermediate between 

those of the parents. However the flower diameter, number of tubular florets, epidermal 

hair height and epidermal hair length exceeded those of both parents. A genomic in situ 

hybridization approach was able to distinguish between the parental genomes in the 

hybrid plant. Among the 54 chromosomes in the putative hybrid, nine were labelled by 

the avidin-FITC assay when the probe comprised genomic DNA of C. chinense, while 

the remaining 45 did not hybridize with the probe. To our knowledge, it is the first report 

of obtainment of intergeneric hybridization between D. crassum (kitam.) kitam. and C. 

chinense (L.) Makino. 

P88 

Fangping Tang (1,2) 

Fadi Chen (1) 

Sumei Chen (1) 

Nianjun Teng (1) 

Weimin Fang (1) 

(1) College of Horticulture, 

Nanjing Agricultural 

University, Nanjing 210095, 

China 

(2) College of Life Science, 

University of Shaoxing, 

Shaoxing 312000, China 

chenfd@njau.edu.cn 

chensm@njau.edu.cn 

Session Posters 161

P89 

Hongzhi Wu (1,2) 

Sixiang Zheng (2) 

Yufen Bi (3) 

Youyong Zhu (2) 

(1) The Gardening 

Department of Horticulture 

College, Yunnan 

Agriculture University 

Kunming 650201, China 

(2) The National Center for 

Agro-biodiversity, Yunnan 

Agricultural University, 


(3) Department of Pasture 

Science, Yunnan 

Agricultural University, 


zsx1966@tom.com 

VARIATION IN RESISTANCE TO FUSARIUM 

OXYSPORUM F.SP. LILII FROM PROGENIES 

DERIVED FROM INDUCED 2N GAMETES IN 

ORIENTAL LILIES 

Fusarium oxysporum f.sp. lilii was isolated and identified from diseased bulb samples of 

oriental lilies collected in main production regions in yunnan province, China. Difference 

in resistance of different oriental lilies to Fusarium oxysporum f.sp. lilii was detected in a 

clonal test, and a high degree of resistance was found in genotype Cai-74. Cai-74 was 

derived from 4x × 2x crosses. The 4x material was derived from somatic doubling diploid 

cultivar, 2x is from sexual triploid which is derived from induced 2n gametes. Through 

karyotype analysis with Stebbins standard it was proved that significant differences were 

found in karyotype of Cai-74 icompared with the susceptible control. By analysis of 

saponins content in lily bulb with spectrophotometry, saponins content in Cai-74 was 

almost 30% higher than that in the susceptible control. This suggest a correlation 

between saponins and resistance to Fusarium oxysporum f.sp. lilii. The analysis of 

saponins of lily bulbs could be used for the evaluation of the resistance to Fusarium 

oxysporum f.sp. lilii in oriental Lilium cultivars. 

162 Session Posters

IN VITRO CULTURE OF CALADIUM BICOLOR 

(AIT.) VENT. ‘THEP SONGSIL’ AND INCIDENCE 

OF VARIENTS 

Explants from the first fully expanded leaf of Caladium bicolor ‘Thep Songsil’ were 

cultured to determine the appropriate concentration of plant growth regulators in 

modified Murashige and Skoog medium for rapid micropropagation and the variation. 

From 11 in 16 combinations of 6-benzyladenine (BA) and -naphthalene acid (NAA) or 

2,4-dichlorophenoxy acetic acid (2,4-D) promoted callus formation. The combination of 

17.76 μM BA with 2.69 μM NAA was the most effective for callus proliferation in C. 

bicolor cv. ‘Thep Songsil’. Subsequently, plantlets were regenerated on MS medium 

containing 11.2 μM BA. The regenerated plantlets from every combination of plant 

growth regulators were randomly grown in greenhouse. From 11 combinations of BA and 

NAA or BA and 2,4-D, the regenerated plants were divided into 8 types. From each 

growth regulator combinations, 3 – 7 types of regenerated plants were observed. The 

occurrence of variants varied from 14.76 – 57.14 percent. The mode and incidence of 

variants were discussed. It was shown that the major influence on variation was not due 

to plant growth regulators in the medium. A few new regenerated variants were different 

from the original plants, potential for commercial value. 

P90 

C. Thepsithar 


K. Obsuwan 







cchockpisit@yahoo.com 

Session Posters 163

P91 


C. Thepsithar 








SOMACLONAL VARIATION OF CALADIUM 

BICOLOR (AIT.) VENT. ‘JAO YING’ FROM IN 

VITRO CULTURE 

In vitro culture of Caladium bicolor by multiple shoot production via callus induction was 

established. Explants from the first fully expanded leaf of C. bicolor cv. ‘Jao Ying’, a 

commercial cultivar, were cultured to determine the appropriate concentrations of plant 

growth regulators in modified Murashige and Skoog medium for rapid micropropagation 

and variation. All 10 combinations of benzyladenine (BA) and -naphthalene acid (NAA) 

provided callus induction. The combination of 8.87 μM BA with 2.69 μM NAA was the 

most effective for callus proliferation. Subsequently, plantlets were regenerated from calli 

on MS medium containing 11.2 μM BA. Most regenerated plants from in vitro culture, 

grown in glasshouse conditions, were more vigorous than the original ones. Within the 

10 combinations of BA and NAA, the regenerated plants could be characterized into 13 

types. From each growth regulator combination, 3 – 7 types of the regenerated plants 

were observed. The occurrence of variants varied from 22.41 – 87.32 percent. It was 

shown that the major influence on variations was not due to the plant growth regulators 

in the medium. The mode and incidence of variations were discussed. Many regenerated 

variants were totally different from the original plants, and tending to great possibility for 

commercial value. 

164 Session Posters

GISH/FISH AS A TOOL TO CHARACTERISE 

HYBRIDS WITH SMALL GENOMES AND 

CHROMOSOMES 

Fluorescence and Genomic in situ hybridisation (FISH and GISH) are useful tools to 

characterise chromosomes of a genotype and to analyse hybrid plants and natural 

polyploids as to their origin, genomic composition and intergenomic rearrangements. 

However, for plants with very small genomes and small chromosomes, GISH and FISH 

are difficult to perform. Visualisation of the morphology of small chromosomes is 

hampered and often only heterochromatin regions are labelled using GISH. In this study 

the in situ hybridisation technology was adapted for woody ornamentals, commonly 

characterised by small genomes, a high amount of small chromosomes and no 

sequence information. 

Firstly, detailed karyotypes of Hydrangea macrophylla, H. paniculata and H. 

quercifolia were constructed on the basis of arm lengths and centromeric index, together 

with 45S rDNA FISH. The variability among 3 species was expressed by chromosome 

morphology and 45S rDNA signals. The chromosome portraits made in this study can be 

used to trace chromosome behaviour in interspecific hybrids resulting from breeding 

work between the 3 species. 

Secondly, GISH was performed on Buddleja and Hibiscus hybrids resulting from an 

interspecific breeding program between different species of both genera. Using B. 

globosa as a probe, GISH analyses on Buddleja x weyeriana (an F2 selection of B. 

globosa x B. davidii) and F1 and F2 hybrids of B. davidii x B. x weyeriana crosses 

proved that all chromatin material of B. globosa was introgressed into the B. davidii 

chromosomes. Also F1 and F2 hybrids of H. syriacus x H. paramutabilis were analysed 

using GISH with H. syriacus as a probe. Also here recombinant chromosomes showing 

introgression of H. syriacus DNA in H. paramutabilis were detected. 

The adaptation on the in situ hybridisation protocol for woody ornamentals consists 

of searching (i) the optimal probe/block ratio (not lower than 1/80) and (ii) the best 

labelling and detection system (biotin versus digoxigenin) for the probe. 

P92 

Katrijn Van Laere (1) 

Ludmila Khrustaleva (2) 

Johan Van 



(1,3) 







(2) Center of Molecular 

Biotechnology, Russian 

State Agricultural 

University, Moscow 

Agricultural Academy; 

Timiryazev Street 49, 

127550 Moscow, Russia 





Gent, Belgium 

katrijn.vanlaere@ilvo. 

vlaanderen.be 

Session Posters 165

P93 

H. Vejsadová (1) 

J. Šedivá (1) 

L. Havel (2) 

H. Vlašínová (2) 

(1) Silva Tarouca Institute 

for Landscape and 

Ornamental Gardening, 

Publ. Res. Inst., 

252 43 Průhonice, Czech 

Republic 

(2) Mendel University of 

Agriculture and Forestry, 

Zemědělská 1, 613 00 

Brno, Czech Republic 

vejsadova@vukoz.cz 

lhavel@mendelu.cz 

ORGANOGENESIS INDUCTION IN RESISTANT 

AND NONRESISTANT HORSE CHESTNUT 

(AESCULUS HIPPOCASTANUM) 

Horse chestnut (Aesculus hippocastanum) belongs to horticulturally valuable woody 

species. Successfulness of its planting in Europe is decreased due to a pest invasion, 

the horse chestnut leaf miner Cameraria ohridella. Research objective of this work was 

organogenesis induction in explants taken from grafted plants of resistant and 

nonresistant horse chestnut individuals. Explants (shoot apex of vegetative buds, petiole 

and shoot segment) collected in winter, summer and autumn season, were cultured on 

WPM and MS medium containing growth regulators in different concentrations. The 

endogenous bacterial contamination was showed to be limiting factor of regeneration 

induction. In spite of application of wide–spectrum antibiotics (Cefalotin, Cefotaxim, 

Timentin a Vancoccin), no viable in vitro culture was derived from winter buds. However 

summer and autumn buds were found as the most suitable and responsive explants with 

the lowest rate of bacterial infection (about 5%); regeneration of these isolated shoot 

apices was higher as compared to petiole and shoot segments. Lower shoot proliferation 

(30%) of autumn than summer explants (50–80%) was demonstrated. The presence of 

cytokinin BA (benzyladenine) in medium was an important factor of organogenesis 

induction. 

166 Session Posters

BREEDING TROPICAL FRUIT CROPS FOR 

ORNAMENTAL PURPOSES 

Embrapa Cassava and Tropical Fruits has many germplasm collections under field 

conditions to give support to breeding programs carried out there. These programs 

were focused just to the development of new materials for fresh fruit consumption or 

processing. A new approach has begun in 2002 looking for ornamental genotypes in 

these gene banks, mainly in the pineapple, banana, citrus and acerola collections. The 

identification, characterization and selection of potential genotypes to be used as potted 

plants, cut flowers and landscape plants have been done. Initially, the morphological 

descriptors specific to each crop developed by IPGRI were adopted. Nevertheless, 

these descriptors are being adapted for use in the ornamental fruit crops aiming at 

cultivar protection and patenting. After the identification of interesting genotypes with 

ornamental potential, several controlled crosses were performed with pineapple, 

banana and citrus, resulting in the production of several promising hybrids. The 

resistance or tolerance against the most important diseases of these fruit crops have 

been considered in this work due to the economic importance they have in Brazil. 

Currently, new hybrids of pineapple, banana, citrus and some acerola genotypes 

identified in the germplasm collection are under field and greenhouse evaluations, 

depending on their intended use. In addition, a very interesting and novelty product was 

also generated: the minifruits. These are small, non-edible fruits whose appearance is 

similar to the regular ones but destined to a differentiated marked. 

P94 

Fernanda Vidigal Duarte 

Souza (1) 

Janay Almeida dos 

Santos-Serejo (1) 

Rogério Ritzinger (1) 

Walter Soares dos Santos 

Filho (1) 

Everton Hilo de Souza (2) 

(1) Embrapa Cassava and 

Tropical Fruits, Caixa Postal 

007, Cruz das Almas, Bahia, 

Brazil 

(2) M.Sc. student of 

Universidade Federal do 

Reconcavo da Bahia. Brazil 

fernanda@cnpmf. 

embrapa.br 

Session Posters 167

P95 

TISSUE CULTURE OF LEAF EXPLANTS OF 

ORIENTAL LILIUM ‘TIBER’ 

Wang Yue 

Tang Dongqin 

Li Weiwei 

Tang Kexuan 

School of Agriculture and 

Biology, Shanghai 

JiaoTong University, 

P.R.China 

wangyue- 

0921@sjtu.edu.cn 

Young leaves of Oriental Lilium ‘Tiber’ were divided into petiole and lamina, respectively 

as explant materials in tissue culture. The results showed that in MS media with different 

concentrations of NAA and BA, the differentiation ability of the petiole was stronger than 

that of the lamina. The combination of NAA and BA results petiole differentiation, while 

and the increase of BA reduced root formation. For the petiole, MS+NAA2.0mg.l - 

1 +BA0.10mg.l -1 was the best medium to induce shoots. MS+NAA0.50 and 2.0 mg.l - 

1 +BA2.0 mg.l -1 induced yellow and compact callus with successive shoot differentiation. 

The most appropriate medium for root induction was 1/2MS+NAA0.20 mg.l -1 . The 

concentration of sucrose in the medium affected the weight increase of the bulblets. MS 

media with every concentration of sucrose could make weight increase of the bulblets, 

however MS medium plus 60g.l -1 sucrose showed the highest increase. The results 

suggested that the leaf segments could be a good alternative as explants, which can 

provide material for physiological and molecular (breeding) research. . 

168 Session Posters

RAPD MARKERS DEVELOPED FOR 

IDENTIFYING MALE STERILITY IN TAGETES 

ERECTA 

Various experimental protocols for RAPD analysis of genes related to fertility in Tagetes 

erecta were systematically studied and an optimum program was developed for setting 

up a stable system. 

CTAB method and SDS method were compared to extract DNA from different tissues 

of Tagetes erecta, the quality of DNA extracted was evaluated by spectrophotometry and 

PCR. Results showed that total DNA extracted from young leaves by CTAB method was 

suitable for the analysis of RAPD. The concentration of DNA template, MgCl2, buffer 

and dNTPs were examined and the procedure was redesigned for setting up the RAPD 

program. Amplifications were then carried out in 20 µL reactions containing 5µL diluted 

DNA template, 2 µL 10× PCR buffer, 2.5 mmol MgCl 2 , 0.3 mmol dNTP, 0.3μmol primer 

and 1.50U Taq DNA polymerase. PCR reaction were operated under the following 

program conditions: 5 min at 94 before amplification, 40 cycles of 94 for 30 s, 37 1 

min, 72 1 min, followed by one cycle of 72 for 1 min. 

By using this sytem, sterile and fertile plants of W205, a genic male sterile line in 

Tagetes erecta were tested. 288 bands were obtained from 64 RAPD primers screened, 

among which a band of 980bp derived from the primer G-02(5’-GGCACTGAGG-3’) was 

coseparated with fertile plants. This RAPD marker named as G-02-980 could be used to 

identify male and fertile seedlings. 

P96 

Wang Zhi-gang 

Wang Ping 

Yin Dong-sheng 

Pan Bai-tao 

Zhang Hui-hua 

Institute of Floriculture, 

Liaoning Academy of 

Agricultural Sciences, 

Shenyang 110161, China 

wangzga@yahoo.com.cn 

Session Posters 169

P97 

Zhuhua Wu 

Mengli Xi 

Jisen Shi 

The Key Laboratory of 

Forest Genetics and 

Biotechnology of State 

Education Administration of 

China, at Nanjing Forestry 

University, Nanjing China, 

210037 

jshi@njfu.edu.cn 

GENETIC DIVERSITY AND SPATIAL 

AUTOCORRELATION OF GENETIC 

STRUCTURE OF LILIUM REGALE 

Lilium regale is a narrow distribute endemic specie in China. It made an important 

contribution to lily breeding. L. regale grows at dry valley in southwestern of China. It had 

been continually destroyed by human and earthquakes. To protect the species is quite 

pressing. However, there was seldom research on its genetic structure in past years. We 

research on the genetic diversity and spatial autocorrelation of genetic structure of L. 

regale base on ISSR molecular marker. The results indicated that, at the species level, 

the proportion of polymorphic loci was 97.7%, the effective number of allele (Ne) were 

1.994 4, Shannon diversity index (I) were 0.3339. At the population level, the average 

expected heterozygosity was 0.664 0.Shannon diversity index (I) were 0.272 0.They 

showed that genetic diversity of L. regale was high. The result by AMOVA analysis 

indicated that the variation within population account 83.8% and variation among 

populations accounted for 11.849%, and the gene flow was 2.588. It demonstrated that 

the relationship of populations was closer. Gene differentiation was acute within 

population. 

We analyzed the spatial autocorrelation of genetic structure of 4 populations with 

more than 35 samples. The Moran’s I correlograms revealed no significant spatial 

structure within the 4 populations. It indicated that genetic variations of the most 

polymorphic loci within these populations were randomly distributed.However, there 

were gaps at the distance of 3~4m,5~6m and 8~10 m , and there were intrusion at a 

little polymorphic loci. 

Base on these results, we regarded that the conservation approach of L. regale were 

to decrease disturbances, to preserve unique genetic variation , pay attention to 

individuals grown at the distance of 3-4m, 5-6 and 8-10m , to create lord to increase 

gene flow. 

170 Session Posters

GENOME COMPOSITION OF INTERSPECIFIC 

HYBRIDS AMONG THREE GENOMES IN 

LILIUM: A MULTICOLOR GENOMIC IN SITU 

HYBRIDIZATION ANALYSIS 

In Lilium, the Longiflorum (L) hybrids have trumpet-shaped white flowers with distinctive 

fragrance and year round forcing ability, the Oriental hybrids (O) have mostly large pink 

and white flowers with a sweet fragrance and are resistant to Botrytis elliptica, the 

Trumpet hybrids (T) have trumpet-shape, pink or yellow flowers with a strong growth and 

are resistant to Fusarium. With the aim of combining the valuable traits of the three 

different genomes, crosses were made between a triploid LLO-hybrid and a tetraploid 

LT-hybrid. The tetraploid LT-hybrid originated from somatic doubling of a diploid F1- 

hybrid. From this cross, 71 plants were resulted from embryo rescue. 29 of them were 

tested for DNA-content using flowcytometry. Mc GISH (Multicolor Genomic In Situ 

Hybridization) was used to analyze these progenies. The results showed that: 

1) the ploidy level of these progenies, based on DNA-content varied from almost 

triploid to tetraploid with a ploidy level from 3.1 to 3.8, and the chromosome number 

varied from 40 to 46. 

2) the aneuploid progenies possessed chromosomes from all three genomes, 

indicating the hybrid character of the hybrids. 

3) the hybrids owned 24 chromosomes of the Longiflorum genome, 12 chromosomes 

of the Trumpet genome and 4-8 originated from Oriental genome. 

4) several genotypes showed the presence of 1 or 2 so-called B chromosomes. 

P98 

Songlin Xie (1) 

Nadeem Khan (2) 

M.S.Ramanna (2) 


(1) College of Horticulture, 

Northwest A&F Univ., 

Yangling, Shaanxi 712100, 

P.R. China 

(2) Wageningen University 

and Research Centre, Plant 

Breeding P.O. Box 386, 

Wageningen 7600 AJ, The 

Netherlands 

songlin.xie@wur.nl 

Session Posters 171

P99 

Yesim Yalcin-Mendi (1) 

Selay Eldogan (1) 

Ceren Unek (1) 

Ayfer Torun (2) 

Leyla Hizarci (3) 

Oya Işik (3) 


Horticulture, Faculty of 

Agriculture, Univ. of 

Cukurova, Balcali, Adana 

(2) Department of Soil 

Science, Faculty of 




Aquaculture, Faculty of 



yesimcan@cu.edu.tr 

THE USAGE OF SPIRULINA ON 

ACCLIMATIZATION OF MICROPROPAGATED 

BEGONIA (B. SEMPERFLORENS) PLANTLETS 

Spirulina (Arthrospira platensis) is blue-green micro-algae. The usage of algae on 

acclimatization allows plants to tolerate greater levels of cold, reduce the amount of 

damage and take nutrient elements. 

Micropropagated begonia plants acclimatized with Spirulina showed a greater leaf 

length and leaf width than the control plants without Spirulina. The leaf lengths in control 

plants and the plants with spiriluna were 2.63 and 2.97 cm, respectively. In addition, the 

leaf widths in the control plants and the plants with spiriluna were 1.93 and 2.1 cm, 

respectively. 

Macro nutrient elements tested in control plants and the plants with spirulina showed 

similar results. However, Fe (108 mg kg -1 ) and Zn (132 mg kg -1 ) concentrations in control 

plants were lower than Fe (189 mg kg -1 ) and Zn (186 mg kg -1 ) concentrations in the 

plants acclimatized with Spirulina. 

172 Session Posters

8. List of authors 

Abdullah, S. 

P86 

Bridgen, M. 

L10 

Abe, Y. 

P64 P67 

Buddharak, P. 

L32 

Abel, S. 

P46 

Burge, G.K. 

L14 

Ahn, I.S. 

LP6 P35 

Bushehri, A.-A. 

L4 

Aida, R. 

LP17 P65 P81 P83 

Caser, M 

LP2 

Alemán, M.M. 

P25 P26 

Cassano-Montebello, C. 

P3 

Allavena, A. 

LP18 L35 

Cassetti, A. 

L35 

Amaki, W. 

P1 

Castro, A.C.R. 

P15 P16 

Amiri, R. 

P33 

Chabannes, M. 

L36 

Anderson, N.O. 

L24 P2 

Chandej, R. 

P55 

Anuntalabhochai, S. 

LP10 LP15 P55 

Chandler, S. 

L6 

Aragao, F.A.S. 

P16 

Chen, F. 

P88 

Arab, M. 

P4 

Chen, S. 

P88 

Araki, H. 

P42 

Chen, W-H. 

P38 

Aranda-Peres, A. 

P3 

Cheng, F. 

P17 

Ardelean, M. 

P5 

Chikuo, Y. 

P57 

Areekijseree, M. 

P6 

Chin, D.P. 

LP11 P18 P70 

Arens, P. 

LP2 L16 L37 P7 

P76 

Cho, H.R. 

Cho, H.S. 

P78 

P71 

Astarini, I.A. 

P8 

Choi, J.-C. 

LP14 P19 

Auvray, G. 

L13 

Christensen, B. 

LP12 

Awal, A. 

P86 

Christiaens, A. 

P20 

Azadi, P. 

LP11 

Chundet, R. 

L32 

Azlina-Hasbullah, N. 

P86 

Chung, J-D. 

LP6 P36 

Babaei, A. 

P33 

Chung, Y-S. 

P51 

Bagheri, A. 

P41 

Cordea, M. 

P5 

Bai-tao, P. 

P96 

Czernicka, M. 

LP3 

Baliuniene, A. 

P40 

Dapkuniene, S. 

P40 P84 

Ballardini. M. 

LP13 P30 

Davarynejad, G.H. 

P41 

Balode, A. 

L18 

De Benedetti, L. 

LP13 P14 P30 

Barba-Gonzalez, R. 

LP1 

De Carvalho, A.C.P.P. 

P15 P16 

Barche, S. 

P9 P45 

De Keyser, E. 

L7 P20 

Basaki, T. 

LP16 

De Ponti, O. 

L2 

Bertsouklis, K.F. 

P10 

De Riek, J. 

L7 L37 P20 

Bi, J. 

P52 

De Souza, E.H. 

P94 

Bi, Y. 

P89 

De Vries, D.P. 

L25 P21 

Bianchini, C. 

LP13 

Debener, T. 

L15 

Bijman, P.J.J. 

P11 

Debenham, M. 

L14 

Biniari, K. 

P10 

Demmink, J.F. 

P11 

Binti, F. 

P12 

Dewitte, A. 

P22 

Blokland, J 

L20 

Dhooghe, E. 

LP4 

Bohanec, B. 

P62 

Dhyani, A. 

P23 

Borchert, T. 

P13 

Dianati, S. 

P4 P33 

Borghi, C. 

LP18 L35 

Dobrochowska, M. 

P50 

Borja, M. 

LP5 L36 

Doi, H. 

P24 

Braglia, L. 

LP13 P14 

Dolgov, S.V. 

L31 

Braun. H-P. 

P79 

Dolstra, O. 

L17 

List of authors 173

Dongqin, T. 

P95 

Hiratsu, K. 

LP17 P65 

Dong-sheng,Y. 

P96 

Hizarci, L. 

P99 

Dos Santos Filho, W.S. 

P94 

Hof, L. 

P7 

Dos Santos-Serejo, J.A. 

P94 

Hohe, A. 

P13 

Dubois, L.A.M. 

L25 P21 

Hop. M.E.C.M. 

P34 

Eeckhaut, T. 

L11 P27 

Horikawa, M. 

P87 

Eldogan, S. 

P99 

Hoshino, Y. 

P42 

Endo, R. 

LP17 

Hui-hua, Z. 

P96 

Engel, J. 

P28 

Hwang, Y.J. 

LP6 P35 P36 

Esselink, D. 

L4 L22 P7 

Ichihashi, S. 

P29 

Etcheverry, A. 

P25 P26 

Ikeda, K. 

LP17 

Ezura, H. 

L34 

Ikeda, M. 

LP17 P43 

Faber, D. 

L1 

Imhof, L. 

LP5 

Facciuto, G. 

LP5 

Ishihara, M. 

P47 

Fang, W. 

P88 

Ishikawa, R. 

P43 

Feng, Y. 

L5 

Isik, O. 

P99 

Fleming, T.F. 

P26 

Isuzugawa, K 

LP17 

Frick, J. 

P2 

Jafarkhani Kermani, M. 

LP16 P41 

Fujita, Y-I. 

P85 

Jajarmi-Islamic, V. 

P37 

Fukuda, N. 

L34 

Jancys, S. 

P40 

Futagami, Y. 

P29 

Janssen, A. 

LP20 

Gâteblé, G. 

L12 

Javadi, H. 

P77 

Geibel, M. 

P28 

Jean, G-T. 

P38 

Ghaffari, M.R. 

LP16 

Jensen, E,B. 

LP12 

Gi, G-y. 

LP14 P19 

Jeong, B.J. 

P71 

Gitonga, V.W. 

L8 L17 

Jevremovic, S 

P39 

Giovannini, A. 

LP19 P14 P30 

Jo, Y.S. 

P71 

Gobin, B. 

P20 

Joung, Y-H. 

LP14 P19 P51 

Gómez, C. 

P25 P26 

Juodkaite, R. 

P40 P84 

Greppi, J.A. 

P31 

Kakizaki, Y. 

P64 

Grzebelus, D. 

LP3 

Kama, S. 

P29 

Grzebelus, E. 

LP3 

Kang, S.Y. 

LP6 P35 P51 P71 

Guerrero, M.E. 

P76 

Kao, Y-L. 

P38 

Haghnazari, A. 

LP16 

Kapusta, V. 

L13 

Haghighi, A.R. 

P77 

Karimiani, Z.G. 

P41 

Hagiwara, J.C. 

P31 

Kashihara, Y. 

P42 

Hammer, K. 

P48 

Kato, J. 

P29 P43 

Han, T.-H. 

LP14 P19 P51 

Kexuan, T. 

P95 

Han, Y.Y. 

P36 

Khalighi, A. 

L4 

Handa, T. 

LP15 L34 

Khan, N. 

P44 P98 

Hardini, Y. 

P8 

Khan, R.S. 

LP11 

Havel, L. 

P93 

Khoshbakht, K. 

P48 

Hayashi, M. 

P43 

Khrustaleva, L. 

P92 

Heidarhaee, R. 

P33 

Kim, K.W. 

P36 

Hieber, D.A. 

L19 

Kim, S.T. 

P19 P36 

Hikage, T. 

P24 

Kim, W.H. 

LP14 P36 

Hirano, T. 

P42 

Kingman, P. 

P76 

174 List of authors

Kirad, K.S. 

P9 P45 

Mat Taha, R. 

P86 

Kishor, R.K. 

LP8 

Matsui, K. 

LP17 

Klein, M. 

LP3 

Matsushita, Y. 

P57 

Kleizaite, V. 

P73 

Mattaha, R. 

P32 

Kleynhans, R. 

L23 

Maurer, A. 

LP20 

Klocke, E. 

P46 

Maziah, M. 

P58 

Kobayashi, N. 

P47 

Meiners, J. 

P59 

Kodde, L. 

P7 

Mercuri, A. 

LP13 P14 P30 

Kollman, E. 

Komatsu, T. 

L10 

P81 

Mii, M. 

L9 LP11 P18 P29 

P43 P61 P70 

Koning-Boucoiran, C.F.S. 

L8 L17 

Miotani, M. 

P61 

Koobaz, P. 

LP16 

Mishiba, K-i. 

P43 

Koyama, T. 

LP17 

Mitiouchkina, T.Yu. 

L31 

Korbin, M. 

P74 

Mitsuda, N. 

LP17 P65 P83 

Krämer, R. 

P66 P72 

Miyoshi, K. 

P82 

Krens, F.A. 

L8 L17 L33 

Mod-Azzeme, A. 

P58 

Krieger, E. 

P49 

Modi, M. 

P45 

Kristiansen, K. 

L26 

Mohamed, N. 

P60 

Krueger, K. 

P13 

Morais, E.B. 

P15 

Krupa-Malkiewicz, M. 

P80 

Morgan, E.R. 

L14 

Kuehnle, A.R. 

L19 

Morita, Y. 

P61 

Kulpa, D. 

P50 

Mourao, I.C.S. 

P15 P16 

Kuras, A. 

P74 

Mozaffarian, V. 

L4 

Kuroda, K. 

LP11 

Mudalige-Jayawickrama, R.G. 

L19 

Kwon, M.K. 

P36 

Müller, R. 

LP12 

Laura, M. 

LP18 L35 

Murovec, J. 

P62 

Lee, G.J. 

P51 P71 

Murata, N. 

P42 

Lee, J-H. 

P51 

Naderi, R. 

L4 P63 

Lemay, V. 

L12 

Naing, A.H. 

P35 

Leus, L. 

L28 LP19 

Nakamura, I. 

P29 P43 

Levko, G.D. 

P53 

Nakashima, K. 

P29 

Lim, K.B. 

LP6 P35 P36 

Nakatsuka, T. 

LP17 P64 P67 

Lin, C-Y. 

P54 

Nakatsuka, A. 

P47 

Liu, Q. 

L5 

Nambiar, N. 

P58 

Liu, T-H. 

P54 

Nanakorn, W. 

LP10 LP15 

Loges, V. 

P15 

Narumi, T. 

LP17 P65 P83 

Lootens, P. 

L7 

Nautiyal, B.P. 

P23 

Lorenzen, B. 

L3 

Nautiyal, M.C. 

P23 

Lu, C. 

L10 

Nehrlich, S. 

P66 

Lux, S. 

L29 

Niedoba, K. 

P74 

Macquaire-Le Pocreau, N. 

L13 

Niki, T. 

P65 P81 

Mahadtanapuk, S. 

LP10 LP15 P55 

Nishihara, M. 

LP17 P64 P67 

Malécot, V. 

L13 

Nishijima, T. 

P65 P81 

Marasek-Ciolakowska, A. 

LP7 P56 P76 

Noordijk, Y. 

P7 

Marazuela, E. 

L36 

Norouzi, P. 

P63 

Mardi, M. 

LP16 

Nowicka, A. 

LP3 

Martìn, D. 

P25 

Obsuwan, K. 

L19 P68 P90 

List of authors 175

Ohme-Takagi, M. 

LP17 P65 P81 P83 

Rostoks, N. 

LP9 

Ohmiya, A. 

P85 

Rzepka-Plevnes, D. 

P80 

Ohsawa, R. 

P43 

Saito, K. 

LP17 

Ohtani, M. 

P47 

Saleh Shanjani, P. 

LP16 

Ohtsubo, N. 

LP17 P65 P81 P83 

Sakai, T. 

LP17 

Okazaki, K 

P82 

Samiei, L. 

L4 

Ono, M. 

LP17 

Sanguansermsri, M. 

LP10 LP15 P55 

Onozaki, T. 

P85 

Sasaki, K. 

P81 P83 

Opitz, E. 

L24 

Sato, T 

P82 

Orlikowska, T. 

P69 

Scariot, V. 

LP2 LP19 

Otani, Y. 

P70 

Sediva, J. 

P93 

Ounémoa, J. 

L12 

Shahin, A. 

L16 

Papafotiou, M. 

P10 

Shamsi, S. 

P58 

Park, I.S. 

LP6 P35 

Sharma, G.J. 

LP8 

Park, J.O. 

P71 

Shchennikova, A.V. 

L31 

Park, M.Y. 

P71 

Shi, J. 

P97 

Park, S.K. 

P35 

Shibata, M. 

P57 P85 

Pauwels, E. 

P20 

Shikata, M. 

LP17 P81 P83 

Peccenini, S. 

P30 

Shimada, N. 

P64 

Peres, L.E.P. 

L27 

Shinoda, K. 

P42 

Pérez, S. 

P25 

Shiota, H. 

P29 

Petersen, K.K. 

L26 

Shulga, O.A. 

L31 

Petric, M. 

P39 

Shulian, Y. 

P52 

Ping, W. 

P96 

Singh, D.B. 

P9 P45 

Pinheiro-Martinelli, A. 

P3 

Sirisawat, S. 

L34 

Pipino, L. 

LP19 P14 

Skridaila, A. 

P40 P84 

Pirseyedi, S.M. 

LP16 

Skryabin, K.G. 

L31 

Plaschil, S. 

P66 P72 

Smolik, M. 

P50 

Podwyszynska, M. 

P74 

Smulders, M.J.M. 

L4 L22 

Ponce, F. 

P76 

Somkanae, U. 

P6 P68 

Proscevicius, J. 

P73 

Souza, F.V.D. 

P94 

Pui, D-A. 

P5 

Sparinska, A. 

LP9 

Pyck, N. 

P75 

Sriskandarajah, S. 

LP12 

Quinones, A.M. 

P76 

Stolker, R. 

L8 

Rahiman, A. 

P12 

Stukeniene, G. 

P40 P84 

Rahimi, V. 

P4 

Subotic, A. 

P39 

Ramanna, M.S. 

LP7 P44 P56 P98 

Suh, D.H. 

P35 

Rashidi, V. 

P77 

Sumitomo, K. 

P57 P85 

Regis, C. 

LP18 L35 

Taha, R.M. 

P12 P60 

Reheul, D. 

LP4 

Takahashi, R. 

P24 

Rhee, H.K. 

P78 

Takahata, Y. 

P24 

Ribot, S. 

L8 

Takamura, T. 

P87 

Richter, K. 

P28 

Takeuchi, S 

P1 

Rieksta, D. 

LP9 

Tanaka, N. 

P43 

Ritzinger, R. 

P94 

Tanaka, Y. 

L6 LP17 

Rode, C. 

P79 

Tang, C-Y. 

P38 

Roetenberg, P. 

L30 

Tang, F. 

P88 

176 List of authors

Teng, N. 

P88 

Xie, S. 

P98 

Ter Laak, W. 

P56 

Xi, M. 

P97 

Tera-arusiri, W. 

LP10 

Yalcin, F. 

LP20 

Terakawa, T. 

LP17 

Yalcin-Mendi, Y. 

P99 

Thepsithar, C. 

P6 P68 P90 P91 

Yamagata, A. 

P85 

Thongpukdee, A. 

P6 P68 P90 P91 

Yamaguchi, H. 

LP17 P81 P83 

Timmerman-Vaughan, G. 

L14 

Yamamura, S. 

P67 

Torun, A. 

P99 

Yan, L. 

P52 

Trifunovic, M. 

P39 

Yeh, D-M. 

P54 

Umemura, Y. 

LP17 

Yokoi, S. 

P24 

Unek, C. 

P99 

Yoo, Y-k. 

P51 

Van Bockstaele, E. 

L7 P22 P92 

Yoshida, Y. 

P43 

Van den Hurk, A. 

L2 

Yoshimura, N. 

P87 

Van der Linden, C.G. 

L17 L37 

Yoshioka, S. 

P85 

Van der Schoot, J. 

L17 

Younis, A. 

L24 P2 

Van der Vossen. E. 

LP20 

Yousefzadeh, M. 

P77 

Van Eijk, M. 

LP20 

Yu, H. 

L5 

Van Heusden, A.W. 

L16 L37 

Yue, W. 

P95 

Van Huylenbroeck, J. 

L11 L28 P22 P27 

P75 P92 

Zajac, K. 

Zarif, I. 

P80 

P4 

Van Kronenburg, B.C.E. 

L33 

Zarina, R. 

LP9 

Van Labeke, M.-C. 

L11 LP4 LP19 P20 

Zawadzka, M. 

P69 

Van Laere, K. 

L28 P92 

Zhang, D. 

P17 

Van Liere, H 

LP20 

Zheng, S. 

P89 

Van Orsouw, N. 

LP20 

Zhou, S. 

P17 

Van Tuyl, J.M. 

LP7 L16 P11 P44 

P56 P76 P98 

Zhu, Y. 

Zilinskaite, S. 

P89 

P40 

Vanstechelman, I. 

L11 

Vansteenkiste, H. 

L11 

Vejsadova, H. 

P93 

Veveris, R. 

LP9 

Visser, R.G.F. 

P44 

Vlasinova, H. 

P93 

Vogel, S. 

P26 

Voorrips, R.E. 

L22 

Vosman, B. 

L21 L22 L37 P7 

Wang, Z. 

P96 

Wang, C. 

P52 

Wati-Hareon, N. 

P32 

Weiwei, L. 

P95 

Weizhen, G. 

P52 

Wietsma, W. 

P7 

Winkelmann, T. 

P59 P79 

Witsenboer, H. 

LP20 L37 

Wu, H. 

P89 

Wu, Z. 

P97 

List of authors 177

178

9. List of participants 

(alphabetical order) 

Abdul Rahiman, Fatima 

Institute of Science Biology, University of Malaya 

50603 Kuala Lumpur 

Malaysia 

fatimah_rahiman@hotmail.com 

60379677121 

60379674372 

Amaki, Wakanori 

1737 Funako 

243-0034 Atsugi 

Japan 

amaki@nodai.ac.jp 

81-46-270-6553 

81-46-270-6553 

Abdullah, Sakinah 

Institute of Biological Sciences, Faculty of Science, University of 

Malaya 


Malaysia 

raihani_84@yahoo.com 

60379677121 

60379674178 

Amrad, Avichai 

The Hebrew University of Jerusalem 

76100 Rehovot 

Israel 

amrad@agri.huji.ac.il 

972-52-4404440 

972-8-3489943 

Agathe, Le Gloanic 

Les Fontaines De L'aunay 

49250 Beaufort En Vallee 

France 

pascal.papillon@pepinieres-minier.fr 

02 41 79 48 23 

Anderson, Neil 

Dept. of Horticultural Science, 1970 Folwell Avenue 

55108 Saint Paul, MN 

United States 

ander044@umn.edu 

+1-612-624-6701 

+1-612-624-4941 

Agbonavbare, Paul Ovie 

25 Bola Ige Crescent Bodija 

00234 Ibadan 

Nigeria 

folii2003@yahoo.com 

+2348054064823 

+234288975566 

Ardelean, Marin 

3-5 Manastur St. 

400372 Cluj-Napoca 

Romania 

mardelean@usamvcluj.ro 

+40 264 596384 

+40 264 593792 

Allavena, Andrea 

C.so Inglesi, 508 

18038 Sanremo 

Italy 


+39 0184 694824 

+39 0184 694856 

Areekijseree, Mayuva 

6 Rachamucha-ni Rd., Prapathom Chedi District, Muang 

73000 Nakhon Pathom 

Thailand 

maijackee@yahoo.com 

+66 3425 5093 

+66 3425 5820 

Allavena, Giulia 

Via Ughetto G., 20 

18030 Ventimiglia (IMPERIA) 

Italy 

giulia.allavena01@universitadipavia.it 

(0039) 329 2228663 

Arens, Paul 

Droevendaalsesteeg 1 

6708 PB Wageningen 

Netherlands 

paul.arens@wur.nl 

+31 317 48 08 27 

List of participants 179

Astarini, Ida Ayu 

Biology Department, Faculty of Natural and Agricultural Sciences 

80364 Denpasar 

Indonesia 

idaastarini@yahoo.com 

+6281337174509 

Barba-Gonzalez, Rodrigo 

Av. Normalistas #800 Colinas de la Normal 

44270 Guadalajara, Jalisco 

Mexico 

rbarba@ciatej.net.mx 

+52 (33) 33455200 ext 1701 

+52 (33) 33455245 

Auvray, Gaëlle 

2 rue le Nôtre 

49045 Angers Cedex 01 

France 

gaelle.auvray@angers.inra.fr 

332 41 22 57 83 

332 41 22 54 78 

Bartsch, Melanie 

Herrenhaeuser Str. 2 

30419 Hannover 

Germany 

bartsch@baum.uni-hannover.de 

0049 511 762 4017 

0049 511 762 3608 

Azadi, Pejman 

Laboratory of Plant Cell Technology, Graduate School of 

Horticulture, Chiba University 

648 Matsudo 

271-8510 Matsudo 

Japan 

azadip22@yahoo.com 

+81- 473088852 

+81- 473088852 

Babaei, Alireza 

Chamran high way, Gisha Bridge, Tarbiat Modares University 

(TMU) 

P. O. Box: 14115-365 Tehran 

Iran, Islamic Republic Of 

arbabaei@modares.ac.ir 

(98) 9123599020 

(98) 2144196524 

Berruti, Andrea 

Via Leonardo da Vinci, 44 

10095 Grugliasco (Torino) 

Italy 

andrea.berruti@unito.it 

+390116708935 

+390118798 

Bijman, Paul 

Kreekrug 2 

1718 DC Hoogwoud 

Netherlands 

paulbijman@gmail.com 

06-51260463 

Bahcevandziev, Kiril 

Escola Superior Agraria, Polytechnics of Coimbra, Bencanta 

3040-316 Coimbra 

Portugal 

kiril@esac.pt 

+351/962358100 

Blokland, Judith 

Postbus 462 

2800 AL Gouda 

Netherlands 

j.blokland@plantum.nl 

+31 182 - 68 86 68 

Balode, Antra 

Liela 2 

LV 3001 Jelgava 

Latvia 

antra@ram.lv 

+371 63005629 

=37163005679 

Bogers, Robert 

International Society for Horticultural Science 

6721 GM Bennekom 

Netherlands 

rjbogers.ishs@kpnplanet.nl 

+31 318 420315 

180 List of participants

Bohanec, Borut 

Jamnikarjeve 101 

1000 Ljubljana 

Slovenia 

borut.bohanec@bf.uni-lj.si 

+38614231161 

+38614231088 

Brandwagt, Bas 

Lavendelweg 15 

1435 EW Rijsenhout 

Netherlands 

b.brandwagt@royalvanzanten.com 

+31-297-387113 

+31-297-387070 

Borchert, Thomas 

Kuehnhaeuser Str. 101 

99189 Erfurt 

Germany 

borchert@erfurt.igzev.de 

+49 36201 785 258 

Bridgen, Mark 

3059 Sound Ave. 

11901 Riverhead, NY 

United States 

mpb27@cornell.edu 

631-727-3595 

631-727-3611 

Borghi, Cristina 

C.so Inglesi 508 

18038 Sanremo 

Italy 

borghicristina@yahoo.it 

+39 0184694824 

+39 0184694856 

Brown, Santiago 

San Javier No.215 y Orellana 

00000 Quito 

Ecuador 

santiago@brownbreeding.com 

+59393880426 

Borja, Marise 

Ciudad Universitaria s/n 

28040 Madrid 

Spain 

imasd@promiva.com 

+34 91 616 6428 

+34 91 616 3337 

Buddharak, Phopgao 


56000 Phayao 

Thailand 

su_buddha@hotmail.com 

66-544-666 

66-544-664 

Botden, Niek 

Johannes Bosboomlaan 11 

6717HH Ede 

Netherlands 

info@hortilink.nl 

+31646191855 

+31842224277 

Cadic, Alain 

42 Rue Georges Morel, BP60057 

49071 Beaucouzé 

France 

alain.cadic@angers.inra.fr 

+33 2 41 22 57 72 

+33 2 41 22 57 55 

Bouty, Ellen 

3 rue Alexandre Fleming 

49066 Angers cedex1 

France 

contact@valinov.fr 

33 (0)2 41 72 11 40 

33 (0)2 41 72 35 67 

Carino, Rose Mary 

MB 160 Puguis, 

2601 La Trinidad, Benguet 

Philippines 

cravegies@yahoo.com 

+639095204654 


Caser, Matteo 



Italy 

matteo.caser@unito.it 

+390116708935 

+390118798 

Chin, DongPoh 

648 Matsudo 

271-8510 Chiba Matsudo-city 

Japan 

dpchin@faculty.chiba-u.jp 

+81-47-308-8854 

Castro, Ana Cecilia 

R. Dra. Sara Mesquita 2270 

60511-100 Fortaleza (Ceará) 

Brazil 


00 55 85 33917277 

00 55 85 33917125 

Christensen, Brian 

Højbakkegård Allé 21 

2630 Taastrup 

Denmark 

brc@agrotech.dk 

(+45) 20185928 

Ceulemans, Moritz 

De Jong Lelies Research BV 

1619 AE Andijk 

Netherlands 

m.ceulemans@dejonglelies.nl 

31228591400 

31228592837 

Czernicka, Malgorzata 

al.Mickiewicza 21 

31-120 Krakow 

Poland 

czernickam@ogr.ar.krakow.pl 

0048126625329 

0048126625266 

Chen, Sumei 

No. 1 Weigang 

210095 Nanjing 

China 

chensm@njau.edu.cn 

+86-25-84399101 

+86-25-84395266 

Dapkuniene, Stase 

Kairenu 43 

LT-10239 Vilnius 

Lithuania 

stase.dapkuniene@gf.vu.lt 

+37052317098 

+37052317098 

Chen, Fadi 

No. 1 Weigang 


China 

chenfd@njau.edu.cn 

+86-25-84395231 

+86-25-84395266 

Debener, Thomas 

Herrenhäuser Str. 2 


Germany 

debener@genetik.uni-hannover.de 

+495117622672 

+4951176219292 

Chen, Wen Huei 

No.700, Kaohsiung University Road,Nan-Tzu District, Kaohsiung 

City 

811 Kaohsiung 

Taiwan, Province Of China 

a08539@nuk.edu.tw 

886-7-5919408 

886-7-5919404 

Demmink, Jan 

Vossenweg 16 

6721BN Bennekom 

Netherlands 

j.demmink1@chello.nl 

0031318415974 


de Bont, Diony 

Takii Europe / Sahin 

1424 PC De Kwakel 

Netherlands 

ddbont@sahin.nl 

06-51238947 

de Ponti, Orlando 

P.O. Box 4005 

6080 AA Haelen 

Netherlands 

o.deponti@nunhems.com 

+31 495 453066 

+31 475 599222 

de Buck-Koning, Karin 

Julianaweg 6A 

1711 RP Hensbroek 

Netherlands 

karindebuck@dekkerchrysanten.nl 

0031-226456060 

0031-226456075 

de Riek, Jan 

Caritasstraat 21 

9090 Melle 

Belgium 

jan.deriek@ilvo.vlaanderen.be 

+ 32 9 272 29 00 

+ 32 9 272 29 01 

de Haas-van Dijk, Marinke 

Hoofdstraat 2 

2678CK De Lier 

Netherlands 

marinke@novanthos.nl 

+31624786123 

de Vries, Dirk 

Postbus 4097 

6710 EB Ede 

Netherlands 


0031615098192 

De Keyser, Ellen 


9090 Melle 

Belgium 

ellen.dekeyser@ilvo.vlaanderen.be 

+ 32 9 272 29 00 

+ 32 9 272 29 01 

Dewitte, Angelo 

Wilgenstraat 32 

8800 Roeselare 

Belgium 

angelo.dewitte@katho.be 

003251232330 

003251228258 

den Besten, Jasper 

HAS Den Bosch 

5200 MA '-Hertogenbosch 

Netherlands 

j.dbesten@hasdn.nl 

++ 31 73 69 23 824 

++ 31 73 69 23 699 

Dhooghe, Emmy 

Coupure links 653 

9000 Gent 

Belgium 

emmy.dhooghe@ugent.be 

0032.9.264.60.77 

0032.9.264.62.25 

den Nijs, Ton 

POB 16 

7600 AA Wageningen 

Netherlands 

ton.dennijs@wur.nl 

317 480856 

Dohm, Andrea 

Klemm & Sohn 

70378 Stuttgart 

Germany 

a.dohm@selectaklemm.de 

00497119532557 

00497119532536 


Doi, Hisako 

3-18-8, Ueda 

020-8550 Morioka 

Japan 

u027019@iwate-u.ac.jp 

+81 19-621-6152 

+81 19-621-6107 

Fereol, Leonidas 

73 lot. Domaine Roches Carrées 

97232 Le Lamentin, Martinique 

France 

leonidas.fereol@cirad.fr 

+596596745193 

Dubois, Lidwien 

Bennekomseweg 176 

6704 AL Wageningen 

Netherlands 

lidwien.dubois@wur.nl 

0031620420428 

Gada, Mariame 

Plant Research International 

6707HV Wageningen 

Netherlands 

mariame.gada@wur.nl 

0623444265 

Eeckhaut, Tom 


9090 Melle 

Belgium 

tom.eeckhaut@ilvo.vlaanderen.be 

+ 32 9 272 29 00 

+ 32 9 272 29 01 

Gargul, Joanna 

Herrenhäuser Str. 2 


Germany 

gargul@zier.uni-hannover.de 

+4917667072201 

+495117622654 

Elke, Steffen 

Kipsdorfer Str. 146 

01279 Dresden 

Germany 

e.steffen@pac-elsner.com 

++49 351 255910 

++49 351 2517494 

Gâteblé, Gildas 

BP 711 

98810 Mont Dore - Nouvelle-Caledonie 

France 

gateble@iac.nc 

687 43 70 75 

687 43 70 16 

Engel, Josefine 

Erwin Baur Straße 27 

06484 Quedlinburg 

Germany 

josefine.engel@jki.bund.de 

03946 47613 

Geerlings, Dave 

Lavendelweg 15 

1435EW Rijsenhout 

Netherlands 

d.geerlings@royalvanzanten.com 

+31 297 387112 

Etcheverry, Angela Virginia 

Avenida Bolivia 

4400 SALTA 

Argentina 

avetcheverry@yahoo.com.ar 

00543874255434 

00543874255455 

Geibel, Martin 

Kipsdorfer Str. 146 

01279 Dresden 

Germany 

dr.m.geibel@pac-elsner.com 

++49 351 255910 

++49 351 2517494 


Giovannini, Annalisa 

corso Inglesi 508 

18038 Sanremo (Imperia) 

Italy 

annalisa.giovannini@entecra.it 

++39-0184-694832 

++39-0184-694856 

Haron, Noorma 


Malaya, Kuala Lumpur, Malaysia 


Malaysia 

noorma@um.edu.my 

60379674352 

60379674178 

Gitonga, Virginia 

Wageningen Campus, Building 107, Droevendaalsesteeg 1 


Netherlands 

virginia.gitonga@wur.nl 

+ 31 317 482508 

Havel, Ladislav 

Zemedelska 1 

613 00 Brno 

Czech Republic 

lhavel@mendelu.cz 

+ 420 545 13 30 16 

+ 420 545 13 30 25 

Guy, Morel 

2565, Rue de Montourey 

83600 Frejus 

France 

heloise.morel@cyclamen.com 

00 33 4 94 19 73 15 

00 33 4 94 19 10 59 

Heimovaara, Sjoukje 

PO Box 377 

1430 AJ Aalsmeer 

Netherlands 

s.heimovaara@royalvanzanten.com 

+31 297 38297110 

Han, Tae-Ho 

Department of Horticulture, Chonnam National University, Young- 

Bong Dong 300 

500-757 Buk-Gu, Gwang-Ju 

Korea, Republic Of 


82-62-530-2066 

82-62-530-2069 

Heins, Christiane 

Petersweg 72 

34346 Hann. Muenden 

Germany 

heike.knobel@benary.de 

+495541700933 

+495541700920 

Han, Li 

Yunan Academa of Agricultural Sciences,Longtou Street, 

Kunming,Yunnan,P.R.China 

650205 Kunming,Yunnan 

China 

lihan528@126.com 

+86 871 5891372 

+86 871 5891602 

Héloise, Morel 

2565, Rue de Montourey 

83600 Frejus 

France 

heloise.morel@cyclamen.com 

00 33 4 94 19 73 15 

00 33 4 94 19 10 59 

Handa, Takashi 

Higashimita 1-1, Tama-ku, Kawasaki 

214-8571 Kanagawa 

Japan 

thanda@isc.meiji.ac.jp 

+81-(0)44-934-7814 

+81-(0)44-934-7814 

Herrera, Jeronimo 

16 Alfred street 

4010 Palmerston North 

New Zealand 

momo_herrera@yahoo.com 

+64211136762 


Hofmann, Birgit 

Binger Strasse 

54457 Gensingen 

Germany 

bhofmann@innovaplant.de 

++496727930168 

Jevremovic, Sladjana 

University of Belgrade, Institute for biological research "Sinisa 

Stankovic" 

11000 Belgrade 

Serbia 

sladja@ibiss.bg.ac.rs 

+381 11 2078 425 

+381 11 2761 433 

Hohe, Annette 

Kuehnhaeuser Strasse 101 

99189 Erfurt 

Germany 

hohe@erfurt.igzev.de 

+49 36201 785 210 

+49 36201 785 250 

Juodkaite, Regina 

Kairenu 43 


Lithuania 

regina.juodkaite@gmail.com 

+37052317933 

+37052317933 

Hoogendijk, Erwin 

Wagendwarspad 4 

1771 RN Wieringerwerf 

Netherlands 

erwin@makbreeding.nl 

0031 612738170 

0031 227 555383 

Kapusta, Veronique 

42 rue Georges Morel ; BP60057 

49071 BEAUCOUZE 

France 

veronique.kapusta@angers.inra.fr 

33 2 41 22 57 80 

33 2 41 22 57 55 

Hop, Margareth 

Postbus 85 

2160 AB Lisse 

Netherlands 

Margareth.Hop@WUR.NL 

+31-252-462183 

+31-252-462100 

Kashihara, Yukiko 

Field Science Center for Northern Biosphere, Hokkaido 

University, Nishi 10, Kita 11, Kita-ku 

060-0811 Sapporo 

Japan 

ikuy06@exfarm.agr.hokudai.ac.jp 

+81-11-706-2854 

Hoshino, Yoichiro 

Kita 11, Nishi 10, Kita-ku 

060-0811 Sapporo 

Japan 

hoshino@fsc.hokudai.ac.jp 

+81-(0)11-706-2857 

+81-(0)11-706-2857 

Kato, Juntaro 

Hirosawa, Igaya, Kariya 

448-8542 Aichi 

Japan 

jkatoh@auecc.aichi-edu.ac.jp 

+81-566-26-2645 

+81-566-26-2645 

Hwang, Yoon-Yung 

1370 Sankyuk-dong, Puk-ku, Department of Horticulture, 

Kyungpook National University 

702-701 Daegu 


kblim@knu.ac.kr 

+82 53 950 5726 

+82 53 950 5722 

Kawashima, Moriya 

Saturnesstraat 1, 

2132 HB Hoofddorp 

Netherlands 

moriyakawashima@gmail.com 

+31 (0)68 32 09 925 


Kingman, Pedro 

Mariano Echeverria 284 

N/A Quito 

Ecuador 

pedrok@hilsea.com.ec 

(593)26035717 

Koning-Boucoiran, Carole 

P. O. Box 16 


Netherlands 

carole.boucoiran@wur.nl 

31 317 480870 

31 317 418094 

Kleynhans, Riana 

Private Bag X293 

0001 Pretoria 

South Africa 

Rkleynhans@arc.agric.za 

+27 12 841 9602 

+27 12 808 0348 

Kos, Joost 

Westeinde 62 

1601 BK Enkhuizen 

Netherlands 

joost.kos@syngenta.com 

+31 288 366 183 

Klocke, Evelyn 

Erwin-Baur-Str. 27 

D-06484 Quedlinburg 

Germany 

evelyn.klocke@jki.bund.de 

+49 3946 47 441 

+49 3946 47 400 

Kratzenberg, Sabine 

Petersweg 72 


Germany 


+495541700933 

+495541700920 

Knaap, van der Koen 

Bagijneland 1 

2691 NC 's-Gravenzande 

Netherlands 

k.vanderknaap@combinationsbv.com 

(+31) (0)6-27013125 

(+31) (0)174417219 

Krens, Frans 

P.O. Box 16 


Netherlands 

frans.krens@wur.nl 

+31 317 480962 

Knoppert, René 

Tjaskerlaan 108 

3052 HS Rotterdam 

Netherlands 

vistaverde@xs4all.nl 

00 31 (010) 4229662 

Krieger, Edgar 

Gänsemarkt 45 

20354 Hamburg 

Germany 

info@ciopora.org 

+49 40 555 63 702 

+49 40 555 63 703 

Kobayashi, Nobuo 

Shimane University 

690-8504 Matsue 

Japan 

nkobayashi@life.shimane-u.ac.jp 

+81-852-32-6506 

+81-852-32-6506 

Kristiansen, Kell 

Kirstinebjergvej 10 

5792 Aarslev 

Denmark 

kell.kristiansen@agrsci.dk 

+4589993345 

+45 89993490 


Laura, Marina 

C.so Inglesi 508 

18038 Sanremo 

Italy 

marynal@tiscali.it 

+39 0184694824 

+39 0184694856 

Liu, Qinglin 

No.2 West Yuanmingyuan Road, Haidian Disreict 

100193 Beijing 

China 

liuql@cau.edu.cn 

8610-62733545 

8610-62733545 

Leus, Leen 


9090 Melle 

Belgium 

leen.leus@ilvo.vlaanderen.be 

+ 32 9 272 29 00 

+ 32 9 272 29 01 

Lokker, Bram 

Wageningen University, Department of Plant Sciences 


Netherlands 

bram.lokker@wur.nl 

+31 317 485551 

+31 317 418094 

Levko, Gennady 

VNIISSOK, p/b Lesnoy Gorodok, 

143080 Odinchovo 

Russian Federation 

flowers@vniissok.ru 

(495)599-2442 

495)599-2277 

Lorenzen, Birte 

Straßenbahnstieg 14 

20251 Hamburg 

Germany 

birte.lorenzen@ciopora.org 

+49 (0)40-555-63-702 

+49 (0)40-555-63-703 

Lim, Ki-Byung 

1370 Sankyuk-dong, Puk-ku, Department of Horticulture, 

Kyungpook National University 

702-701 Daegu 


kblim@knu.ac.kr 

+82 53 950 5726 

+82 53 950 5722 

Lux, Susanne 

Ahornweg 3 

53547 Kasbach-Ohlenberg 

Germany 

susanne.lux@meclux.de 

++49 2644 981 333 

++49 2644 981 334 

Lin, Yan 

No.75 Xuefu road 

050061 Shijiazhuang 

China 

linyan65@yahoo.com.cn 

86-311-7684962 

86-311-6839334 

Madsen, Christian Hald 

Slettensvej 215 

5270 Odense N 

Denmark 

chm@pkm.dk 

+45 66189074 

+45 66187954 

Liu, Tzu-Hsuan 

No. 1, Roosevelt Road Section 4 

106 Taipei 


r95628101@ntu.edu.tw 

+886 2 33664858 # 308 

Mahadtanapuk, Supuk 

Naresuan university 

56000 Phayao 

Thailand 


66-54 466666 

66-54 466663 


Mahmood, Maziah 

Dept of Biochemistry. Fac. of Biotechnology & Biomolecular 

Sciences 

Universiti Putra Malaysia. 43440. Serdang. Selangor 

Malaysia 

maziahm@biotech.upm.edu.my 

6012 387 1277 

603 8943 0913 

Massago, Ndobe Nina 

2/98 Sukhumvit SOI 22, Srijinda Mansion 814,Klontan-Klontey 

10110 Bangkok 

Thailand 

nmassango@yahoo.com 

+66847165355 

Malécot, Valery 

2 rue le Nôtre 

49045 Angers Cedex 01 

France 

valery.malecot@agrocampus-ouest.fr 

332 41 22 55 79 

332 41 22 54 78 

Matsushita, Yosuke 

2-1, Fujimoto, Tsukuba, Ibaraki, 305-8519, Japan 

305-8519, Tsukuba 

Japan 

yousuken@affrc.go.jp 

+81-29-838-6820 

+81-29-838-6842 

Mandiit, Sharon 

6 Eagle Crest Subdivision, Bakakeng, Baguio City 

Block 6, Eagle Crest Subdivision, Bakakeng 

2600 Bagui City 

Philippines 

smandiit@yahoo.com 

+639129629696 

Meddens, Frans 

Roggelseweg 33 

6081 CR Haelen 

frans@meddens-articulture.nl 

00 31 475 597520 

00 31 475 597521 

Mansuino, Andrea 

Strada Villetta 31 

18038 Sanremo 

Italy 

andream@nirpinternational.com 

+39335497028 

Mehring Lemper, Manfred 

Petersweg 72 

34346 Hann. Münden 

Germany 


+495541700933 

+495541700920 

Marasek-Ciolokowska, Agnieszka 

Minderbroedersstraat 12 

5911 BP Venlo 

Netherlands 

agnes.ciolakowki-marasek@wur.nl 

31625356263 

Meiners, Julia 

Am Staudengarten 8 

85354 Freising 

Germany 

julia.meiners@fh-weihenstephan.de 

00498161713365 

Marconi, Irene 

FD 200 Pines Park, KM 4, 

2601 La Trinidad, Benguet 

Philippines 

irenetmaeconi@yahoo.com 

+6374 420 5223 

Mercuri, Antonio 

CRA-FSO Corso Inglesi 508 

18038 Sanremo 

Italy 


+39-0184694846 

+39-0184594856 


Mii, Masahiro 

Graduate School of Horticulture, Chiba University, 648 Matsudo 

271-8510 Matsudo 

Japan 

miim@faculty.chiba-u.jp 

+81 47 308 8852 

+81 47 308 8852 

Müller, Renate 

Hojbakkegaard Alle 13 

2630 Taastrup 

Denmark 

ren@life.ku.dk 

+45 35333534 

+4535333478 

Mitiouchkina, Tatiana 

Nayki pr., 6 

142290 Pushchino 

San Marino 

mitiouchkina@rambler.ru 

+7 4967 731779 

+ 7 4967 330527 

Murovec, Jana 

Jamnikarjeva 101 

1000 Ljubljana 

Slovenia 

jana.murovec@bf.uni-lj.si 

+38614231161 

+38614231088 

Mohamed, Normadiha 

Institute of Science Biology, University of Malaya 


Malaysia 

normadiha@yahoo.com 

60379677121 

60379674372 

Naderi, Roohangiz 

Dept. of Horticulture, Faculty of Agriculture, Tehran University 

31587-77871 Karaj 


rnaderi@ut.ac.ir 

0098 912161 4159 

Morgan, Ed 

Private Bag 11600 

4474 Palmerston North 

New Zealand 

morgane@crop.cri.nz 

+ 64 6 355 6172 

Nakatsuka, Takashi 

22-174-4 Narita 

024-0003 Kitakami, Iwate 

Japan 

nakatuka@ibrc.or.jp 

+81-197-68-2911 

+81-197-68-3881 

Morita, Yuki 

648 Matsudo 

271-8510 Chiba Matsudo-shi 

Japan 

mrt_snow_01@yahoo.co.jp 

+81-47-308-8854 

Nebelmeir, Johannes 

Via Piedimonte 8 

39012 Merano (BZ) 

Italy 

johannes@lazzeri.com 

00393357035391 

00390473246666 

Moucheboeuf, Laetitia 

Houndspool Ashcombe Road Dawlish Devon 

EX7 0QP Dawlish 

United Kingdom 

laetitia@whetmanpinks.com 

01626 863328 

01626 888911 

Nehrlich, Stephanie 


06484 Quedlinburg 

Germany 

stephanie.nehrlich@jki.bund.de 

0049 3946 47480 


Nishihara, Masahiro 

22-174-4 Narita 

024-0003 Kitakami, Iwate 

Japan 

mnishiha@ibrc.or.jp 

81-197-68-2911 

81-197-68-3881 

Oladele, Bamidele Olumide 

25 Bola Ige Crescent Bodija 

00234 Ibadan 

Nigeria 

folii2003@yahoo.com 

+2348054064823 

+234288975566 

Obsuwan, Kullanart 

Department of Biology, Faculty of Science, Silpakorn University 

73000 Nakorn Pathom 

Thailand 


66859338386 

Orlikowska, Teresa 

Pomologiczna 18 

96-100 Skierniewice 

Poland 

Teresa.Orlikowska@insad.pl 

+48 46 8345511 

+ 48 46 8332088 

Obaseki-ebor, Nicholas 

NICKSON GLOBAL RESOURCES LIMITED. 

01234 ikeja 

Nigeria 

obasekiebor@yahoo.com 

+2347067089555 

Otani, Yuko 

648 Matsudo 

271-8510 Chiba Matsudo-city 

Japan 

yu_ukokshta@yahoo.co.jp 

+81-47-308-8854 

Ogunshona, Babajide 

Lugansk National Agrarian University Ukraine 

91008 Lugansk 

Ukraine 

jide_wil20@yahoo.com 

+380933529505 

Papafotiou, Maria 

Iera Odos 75 

118 55 Athens 

Greece 

mpapaf@aua.gr 

0030 210 5294555 

0030 210 5294553 

Ohki, Shizuka 

88-1, Futaomote, Awara-shi 

910-4103 Fukui 

Japan 

ohki@fpu.ac.jp 

+81.776.77.1443 

Peres, Lazaro 

LCB/ESALQ Av Padua Dias, 11 CP 09 

13418-900 Piracicaba 

Brazil 

lazaropp@esalq.usp.br 

55-19-34294052 

Ohtsubo, Norihiro 

2-1 Fujimoto, Tsukuba 

305-8519 Ibaraki 

Japan 

nohtsubo@affrc.go.jp 

+81-(0)29-838-6822 

+81-(0)29-838-6822 

Pesteil, Celine 

1 Gilbert road 

Po19 3NP Chichester 

United Kingdom 

c.pesteil@cleangro.co.uk 

00447772632783 


Peters, Jenny 

KeyGene N.V. 

6708 PW Wageningen 

Netherlands 

jpt@keygene.com 

+31 317 466866 

+31 317 424939 

Quiñones, Ana Maria 

Humboldt 155 Y San Ignacio, Quito, Ecuador 

17300788 Quito 

Ecuador 

anamq@hilsea.com.ec 

00 593 99305091 

00 593 93 302 672 

Pipino, Luca 



Italy 

luca.pipino@yahoo.it 

+393807971425 

+390118798 

Randag, Cecilius 

Bosweg 46b 

1756 CJ 't Zand 

Netherlands 

Carlo@sandegroup.nl 

0031 224 571515 

0031 224 573020 

Plaschil, Sylvia 


D-06484 Quedlinburg 

Germany 

sylvia.plaschil@jki.bund.de 

+49 3946 47 491 

+49 3946 47 400 

Readly, Paul 

301 Natividad Road 

93906 Salinas 

United States 

preadly@takii.com 

0018149338677 

0018314434901 

Podwyszynska, Malgorzata 

Pomologiczna Str. 18 

96-100 Skierniewice 

Poland 

mpodwysz@insad.pl 

48 46 8345519 

48 46 332088 

Rode, Christina 

Herrenhaeuserstr. 2 


Germany 

rode@genetik.uni-hannover.de 

+511-762-3264 

Post, Aike 

P.O.box 77 

2676 ZH Maasdijk 

Netherlands 

ap@deliflor.nl 

+31174526200 

+31174526203 

Roetenberg, Paul 

Platinastraat 100 

7554 NB Hengelo 

Netherlands 

p.roetenberg@frederiqueschoice.com 

+31 (0)74 2551875 

Pyck, Nancy 


9090 Melle 

Belgium 

nancy.pyck@ilvo.vlaanderen.be 

+ 32 9 272 29 00 

+ 32 9 272 29 01 

Royer, Frederic 

31 Avenue Jean Medecin 

06000 Nice 

France 

frederic.royer@doriane.com 

33492478444 

33492478449 


Sacco, Ermanno Domenico 

C. so Inglesi 508 

18038 Sanremo (IM) 

Italy 

sacco.ermanno@tiscali.it 

++390184694823 

++390184694856 

Segers, T.A. 

Aalsmeerderweg 694 

1435 ER Rijsenhout 

Netherlands 

thsegers@preesman.com 

0031297327459 

0031297344118 

Samiei, Leila 

Dept. of Horticulture, Agriculture faculty, Tehran University 

31587-77871 Karaj 


leilisamie@yahoo.com 

0098 915 504 10 33 

0098 261 224 87 21 

Shahin, Arwa 

Kwikstaartweide 24 

6708 LS Wageningen 

Netherlands 

arwa.shahin@wur.nl 

0031 647291374 

Sasaki, Katsutomo 

2-1 Fujimoto 

305-8519 Tsukuba 

Japan 

kattu@affrc.go.jp 

+81-29-838-6822 

+81-29-838-6822 

Shankar, Prabhu 

ILVO-PLANT 

9090 Melle 

Belgium 

prabhu.shankar@ilvo.vlaanderen.be 

+ 32 9 272 29 00 

+ 32 9 272 29 01 

Sato, Takao 

34-1 Genpachizawa Aikawa Yuwa 

010-1231 Akita City Akita Prefecture 

Japan 

satou-takao@pref.akita.lg.jp 

+81-18-881-3318 

+81-18-881-3301 

Sharma, G.J. 

Department of Life Sciences, Manipur University, Canchipur 

Sciences 

795003 Imphal 

India 

gjs1951@rediffmail.com 

+91-9436021107 (M) 

+91-385-2435145 

Scariot, Valentina 



Italy 

valentina.scariot@unito.it 

+390116708932 

+390118798 

SHI, Jisen 

159# Longpan Road 


China 

jshi@njfu.edu.cn 

+86-25-85428948 

+86-25-85428948 

Schoellhorn, Rick 

7421 NW 176th St 

32615 Alachua, Florida 

United States 

rick@provenwinners.com 

352-226-0006 

386-462-0431 

Shikata, Masahito 

2-1 Fujimoto 

305-8519 Tsukuba, Ibaraki 

Japan 

mshikata@affrc.go.jp 

+81-29-838-6822 

+81-29-838-6822 


Shiota, Hiroki 

Hirosawa, Igaya, Kariya 

448-8542 Aichi 

Japan 

jkatoh@auecc.aichi-edu.ac.jp 

+81-566-26-2645 

+81-566-26-2645 

Snijder, Ronald 

1e Loosterweg 1a 

2181BL Hillegom 

Netherlands 

r.snijder@royalvanzanten.com 

+31252535353 

+31252535455 

Shulga, Olga 

prospekt 60-let Oktyabrya 7/1 

117312 Moscow 

Russian Federation 

shulga@biengi.ac.ru 

007-499-135-62-19 

007-499-135-05-71 

Snijders, Charles 

Aalsmeerderweg 943 

1435 EP Rijsenhout 

Netherlands 

csnijders@ballhort.com 

+31618109000 

+31297381365 

Simons, Thijs 

Postbus 462 

2800 AL Gouda 

Netherlands 

t.simons@plantum.nl 

+31 182 - 68 86 68 

Sobek, Renate 

Westeinde 62 


Netherlands 

renate.sobek@syngenta.com 

+31 288 366 104 

Sirisawat, Supatida 

University of Tsukuba, Ichinoya 3-103, Tennodai 2-1 

305-0006 Tsukuba, Ibaraki 

Japan 

supatidas@gmail.com 

81-80-5543-7594 

Sparinska, Anta 

2 Kandavas street 

LV-1083 Riga 

Latvia 

anta.sparinska@lu.lv 

+37129446249 

+37167450852 

Smaal, Andre 

Achterweg 58a 

1424 PR De Kwakel 

Netherlands 

info@agriom.nl 

0297-344078 

0297-344102 

Steentjes, Rudie 

Dorpsstraat 11-B 

1566 AA Assendelft 

Netherlands 

info@hortilink.nl 

075-6873853 

375-6873417 

Smulders, Marinus J.M. 

P.O.Box 16 


Netherlands 

rene.smulders@wur.nl 

+31317480840 

Steur, Nico 

Dorpsstraat 11-B 

1566 AA Assendelft 

Netherlands 

info@bromelia.com 

075-6873853 

375-6873417 


Stukeniene, Gitana 

Kairenu 43 


Lithuania 

stukeniene@gmail.com 

+37052317933 

+37052317933 

Tanaka, Yoshikazu 

Suntory Ltd. 1-1-1 Wakayamadai 

618-8503 Shimamoto 

Japan 

Yoshikazu_Tanaka@suntory.co.jp 

+81-75-962-8807 

+81-75-962-3791 

Sumitomo, Katsuhiko 

2-1 Fujimoto, Tsukuba, Ibaraki 

305-8519 Tsukuba 

Japan 

ksumi87@affrc.go.jp 

+81-29-838-6801 

+81-29-838-6842 

Taquet, Valentin 

Ball R&D, Aalsmeerderweg 943 

1435EP Rijsenhout 

Netherlands 

vtaquet@ballhort.com 

+31612701982 

Taha, Rosna 


Malaya, 


Malaysia 

rosna@um.edu.my 

60379674372 

603-79674173 

ten Pas, Nick 

Petersweg 72 


Germany 


+495541700933 

+495541700920 

Takako, Narumi 

2393 Ikenobe 

761-0795 Miki-cho, Kagawa 

Japan 

tnarumi@ag.kagawa-u.ac.jp 

+81-87-891-3130 

+81-87-891-3130 

ter Laak, W.A. 

Witteveld 59 

2041 GC Zandvoort 

Netherlands 

mark.kuiper@tiscali.nl 

06-50205172 

Takamura, Takejiro 

Kagawa University, 2393 Ikenobe, Miki-cho, Kagawa-ken 

761-0795 Miki-cho, 

Japan 

take@ag.kagawa-u.ac.jp 

+81-87-891-3074 

+81-87-891-3074 

Tera arusiri, Werachai 


56000 Phayoa 

Thailand 

werachai_kig_bu@hotmail.com 

66-54 466666 

66-54 466663 

Tan, Mei Lie 

Oberdorf 9 


Germany 

meilietan@aol.com 

+49-5546-999-408 

+49-5546-999-407 

Theobald, Hendrik 

Binger Strasse 

54457 Gensingen 

Germany 

htheobald@innovaplant.de 

++496727930169 


Thepsithar, Chockpisit 

6 Rachamucha-ni Rd., Prapathom Chedi district, Muang 


Thailand 


+66 3425 5093; +66 89202 2524 

+66 3425 5820 

van den Hurk, Anke 

Plantum NL 

2800 AL Gouda 

Netherlands 

a.vandenhurk@plantum.nl 

(+31) 182 - 68 86 68 

(+31) 182 - 68 86 67 

Thongpukdee, Aree 

6 Rachamucha-ni Rd., Prapathom Chedi District, Muang 


Thailand 

toh25@hotmail.com 

+66 3425 5093 

+66 3425 5820 

van der Geest, Berno 

Postbus 251 

6700 AG Wageningen 

Netherlands 

b.van.der.geest@agripartner.nl 

31645654484 

Trojan, Vaclav 

Zemedelska 1 

613 00 Brno 


vaclav.trojan@seznam.cz 

+ 420 545 13 33 89 

+ 420 545 13 30 25 

van der Helm, Frans 

Postbus 8 

1430 AA Aalsmeer 

Netherlands 

frans.vanderhelm@hilverdakooij.nl 

+31(0)297382038 

+31(0)297382020 

Vainstein, Alexander 

Herzel 

76100 Rehovot 

Israel 

vain@agri.huji.ac.il 

972-8-9489082 

van Dijk, Jan 

Anthuriumweg 14 

2665 KV Bleiswijk 

Netherlands 

wk@anthura.nl 

0031105291919 

0031105291929 

Valkering, Annette 

Julianaweg 6A 


Netherlands 

annettevalkering@dekkerchrysanten.nl 

0031-226456060 

0031-226456075 

van Leeuwen, Idy 

Herdersveld 143 

5665 JN Geldrop 

Netherlands 

ivleeuwen@breedwise.nl 

+31 40 2800536 

van den Haak, Jelle 

Wageningen University 

6701 BS Wageningen 

Netherlands 

Jelle.vandenhaak@wur.nl 

0624240180 

van Liere, Herco 

P.O. Box 216 

6700 AE Wageningen 

Netherlands 

herco.van-liere@keygene.com 

+31317466866 

+31317424939 


van Rooijen, Ad 

meerlandenweg 55 

1187 ZR Amstelveen 

Netherlands 

ad.van.rooijen@deruiter.com 

020 6436516 

020-6433778 

van Kleinwee, Dick 

Westeinde 62 


Netherlands 

dicmkvan.kleinwee@syngenta.com 

+31 288 366 104 

van 't Hoenderdal, Kees 

Julianaweg 6a 


Netherlands 

keesvanthoenderdal@dekkerchrysanten.nl 

+31 226 456060 

+31 226 456075 

Van Labeke, Marie-Christine 

Coupure links 653 

9000 Gent 

Belgium 

mariechristine.vanlabeke@ugent.be 

32 9 264 60 71 

32 9 264 62 25 

van Tuyl, Jaap 

Droevendaalse steeg 1 

6708 PP Wageningen 

Netherlands 

jaap.vantuyl@wur.nl 

+31653362858 

Van Laere, Katrijn 


9090 Melle 

Belgium 

katrijn.vanlaere@ilvo.vlaanderen.be 

+ 32 9 272 29 00 

+ 32 9 272 29 01 

Vejsadová, Hana 

Kvetnove nam. 391 

CZ-25243 Pruhonice 


vejsadova@vukoz.cz 

+420 296 528 111 

+420 267 750 440 

Verlinden, Kathryn 

Westeinde 62 


Netherlands 

kathryn.verlinden@syngenta.com 

+31 288 366 104 

van Heusden, Sjaak 

Droevendaalsesteeg 1 

6708PB Wageningen 

Netherlands 

sjaak.vanheusden@wur.nl 

(+)31317481059 

Vosman, Ben 

POBox 16 


Netherlands 


+31 317 480838 

Van Huylenbroeck, Johan 


9090 Melle 

Belgium 

johan.vanhuylenbroeck@ilvo.vlaanderen.be 

+ 32 9 272 29 00 

+ 32 9 272 29 01 

Winkelmann, Traud 

Herrenhaeuser Str. 2 

D-30419 Hannover 

Germany 

traud.winkelmann@zier.uni-hannover.de 

0049 511 762 3602 

0049 511 762 3608 


Witsenboer, Hanneke 

P.O. Box 216 

6700 AE Wageningen 

Netherlands 

hanneke.witsenboer@keygene.com 

+31317466866 

+31317424939 

Wang, Zhi-gang 

No. 84, Dongling Road, Shenyang City, China 

110161 Shenyang City 

China 

wangzga@yahoo.com.cn 

+86 24 3102 5677 

+86 24 3102 5677 

XI, Mengli 

159# Longpan Road 


China 

ximenglinjfu@126.con 

+86-25-85427319 

+86-25-85427319 

Wilde, Katinka 

Klemm & Sohn 

70378 Stuttgart 

Germany 

k.wilde@selectaklemm.de 

00497119532511 

00497119532536 

Xie, Songlin 

Droevendaalsesteeg 1, 6708 PB, Wageningen, the Netherlands 


songlin.xie@wur.nl 

+31 647229425 

Zhou, Shujun 

Horticulture Department, College of Agriculture and 

Biotechnology, Zhejiang University 

310029 No. 258 of Kaixuan Road, Hangzhou City, Zhejiang 

Province 

zhou2007@zju.edu.cn 

0086-571-86971684; 0086-13858060764 

Yalcin Mendi, Yesim 

Cukurova University, Agricultural Faculty, Horticultural 

Department 

01330 Balcali 

Turkey 

ymendi@gmail.com 

00903223386615 

00903223386615 

Ziv, Meira 

Plant Science institute Faculty of Agriculture 

76100 Rehovot 

Israel 

meira@agri.huji.ac.il 

972 8 9489915 

972 8 9489989 

Yeh, Der-Ming 

No. 1, Roosevelt Road Section 4 

106 Taipei 


dmyeh@ntu.edu.tw 

+886 2 33664858 # 205 

+886 2 33664855 


10. Map of Leiden 

Map of Leiden 199

200

List of participants 23 rd 

EUCARPIA symposium Leiden (additional to the book of abstracts) 

Maaike Baggerman Lavendelweg 15 Rijsenhout Royal van Zanten Netherlands m.baggerman@royalvanzanten.com 

Heleen J.M.Bastiaansen Lavendelweg 15 Rijsenhout Royal Van Zanten Netherlands h.bastiaanssen@royalvanzanten.com 

Martin Beers Dwarsweg 15 De Kwakel Florist de Kwakel b.v. Netherlands martinb@gerbera.com 

MARGHERITA BERUTO VIA CARDUCCI, 12 SANREMO (IM) IStituto Regionale per la Italy beruto@regflor.it 

Jasper den Besten Postbox 90108 '-Hertogenbosch HAS Den Bosch Netherlands j.dbesten@hasdn.nl 

Hanne Bjerg Eskildsen Odensevej 82 Marslev Sakata Ornamentals Europe Denmark Hanne.Bjerg-Eskildsen@sakata-eu.com 

Tobias Braunig Postbus 32 Zwijndrecht Florensis Netherlands directie.secretariaat@florensis.com 

Eloy Boon Middenweg 591b Heerhugowaard Iribov Netherlands info@iribov.nl 

Veronique Cadic x x INRA, CR Angers-Nantes France alain.cadic@angers.inra.fr 

Felix Cooper 247 N 2nd St TIpp City Gardens Alive United States felix@gardensalive.com 

Anurag Dhyani High Altitude Plant Srinagar Garhwal High Altitude Plant India anuragdhyani@gmail.com 

Arie van Diepen Middenweg 591b Heerhugowaard Iribov Netherlands info@iribov.nl 

Andrea Dohm Hanfäcker 10 Stuttgart Klemm & Sohn Germany a.dohm@selectaklemm.de 

Henk Dresselhuys Sluuslaan 17 Middelharnis Fides Netherlands h.dresselhuys@fides.nl 

Carla van Eijk Lavendelweg 15 Rijsenhout Royal van Zanten Netherlands c.vaneijk@royalvanzanten.com 

Doeke Faber Virulypad 27 Leiden International Association Netherlands doekefaber@gmail.com 

Suen-Ying Fung P.O. Box 44 Valkenburg Z-H Van Zanten Cuttings BV Netherlands y.fung@royalvanzanten.com 

Ton Groot Cornelis Kuinweg 28a Andijk Goldsmith Seeds Europe Netherlands t.groot@goldsmithseeds.nl 

Eli Guetta Postbus 24 Oldebroek Eden's Creations Netherlands eli.guetta@edens-creations.eu 

Price Hadas Eliyahu Shamir 8 Mishmar Hasiva Danizger "Dan" Flower Israel hadas_pr@walla.com 

Marita Hovers 168-37-1 Persiaran Georgetown PT Tamora Stekindo Malaysia marita.hovers@gmail.com 

Kitty de Jager Bergerven 45 Horn Plant Pathology Advice Netherlands kdejager@kpnmail.nl 

Sladjana Jevremovic Bulevar despota Belgrade University of Belgrade, Serbia sladja@ibiss.bg.ac.rs 

Silvan Kamstra Rietwijkeroordweg 15 Aalsmeer Bartels Breeding Netherlands silvank@bartelsstek.nl 

Lali Karkarashvili 8 Kargareteli str Tbilisi Dadu & Linky Georgia nikatonic@hotmail.com 

Shuntaro Kimura Bindekildevej 36 Odense SV Global flowers A/S Denmark syuntaro-kimura@takii.co.jp 

Khatuna Kordzadze 8 Kargareteli str Tbilisi Dadu & Linky Georgia nikatonic@hotmail.com 

Mark Kuiper x x Horti Partners Netherlands info@hortilink.nl 

Marleen Kusters Dernhorstlaan 9 Twello Schoneveld Twello BV Netherlands breeding@schoneveld.nl 

Raffaele Langella via Fontanatetta-B.go Civitavecchia Albani Vincenzo e Italy breeding@albani.it 

Veerle Lamote Lichtelarestraat 87 Lochristi Floreac NV Belgium veerle.lamote@microflor.com 

Sabine Lorente Rodriguez Zesstedenweg, 36 Grootebroek Agrocampus Ouest Centre Netherlands sabinelorente@msn.com 

Jose Merced Mejia-Munoz UNIVERSIDAD AUOTONOMA CHAPINGO Universidad Autonoma Mexico jmerced58@hotmail.com 

Nika Meskhi 8 Kargareteli str Tbilisi Dadu & Linky Georgia nikatonic@hotmail.com 

Yutaka Mukai 2994-1 Fujimi machi Takii & C0.,LTD. Japan yutaka-mukai@takii.co.jp 

Federica Nicoletti Corso Inglesi 508 Sanremo (IM) CRA FSO Italy f.nicoletti@istflori.it 

Peter Oenings Muensterstrasse 46 Glandorf HEUGER Germany peter.oenings@heuger.com 

Krit Raemakers nieuwe kanaal weg 7b wageningen Genetwister Netherlands c.j.j.m.raemakers@genetwister.nl 

Janay Serejo Rua Rui Barbosa, 440 Cruz das Almas Embrapa Brazil janay@cnpmf.embrapa.br 

Hak Ki Shin Fluitakker 17 Bennekom Wageningen UR Netherlands hakki.shin@wur.nl 

Freddie Skov-Hundevad Odensevej 82 Marslev Sakata Ornamentals Europe Denmark freddie.skov-hundevad@sakata-eu.com 

Henny van Staaveren Postbus 29 Scharendijke Netherlands hvanstaaveren@hetnet.nl 

Susanne Villemoes Villasmindevej 4 Odense SV Ex-Plant A/S Denmark sv@ex-plant.com 

Arie Vletter Postbus 234 RIJNSBURG Gebr. Vletter & Den Haan Netherlands arie@vletterdenhaan.nl 

Aart van Voorst Lavendelweg 15 Rijsenhout Royal Van Zanten Netherlands a.vanvoorst@royalvanzanten.com 

Mirjana Vukosavljev P.O. Box 16 Wageningen Wageningen UR Plant Serbia mirjana.vukosavljev@wur.nl 

Johan van der Woude Fazantlaan 10 Ermelo Koppe Veredeling B.V. Netherlands johan.vander.woude@mkoppebv.nl 

Hongshi Wu x kunming Agricultural University China hongzhi.wu@wur.nl 

Yesim Yalcin Mendi Cukurova University, Adana University Turkey ymendi@gmail.com

XXIIIrd International Eucarpia symposium, Section Ornamentals ...

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