Book of Abstracts - Institut za ratarstvo i povrtarstvo, Novi Sad...(GenXPro GmbH, Frankfurt, Germany) Local Organising Committee Branko Ćupina (Faculty of Agriculture, Novi Sad,

Second GL-TTP Workshop Integrating Legume Science and Crop Breeding

27 and 28 November 2008, Novi Sad, Serbia

PROGRAMME

Time THURSDAY, 27 NOVEMBER 2008 8.30 – 9.30 Registration

9.30 - 9.45 Opening ceremony and welcome address Chairs: GL-TTP Executive Committee and Borislav Kobiljski

9.30 - 9.45 Petr Smýkal (GL-TTP President), Noel Ellis (GL-TTP Past President), Aleksandar Mikić (2GLTTPW Local Organising Committee), Borislav Kobiljski (BREEDING 08), Diego Rubiales (AEP President)

9.45 - 13.00 Session 1: Achievements and challenges in legume breeding Chairs: Balram Sharma and Paolo Annicchiarico

9.45 - 10.15 Ellis T. H. N.: Genetics and breeding in legumes 10.15 - 10.45 Sharma B.: Present status of genetic research in lentil (Lens culinaris)

10.45 - 11.15 Đorđević V., Miladinović J., Balešević-Tubić S., Đukić V.: Future perspective in soybean breeding

11.15 - 11.30 Coffee break and poster exhibition 11.30 - 12.00 Annicchiarico P., Scotti C.: Some burning questions for lucerne breeders 12.00 - 12.30 Kovačević B., Orlović S., Pilipović A.: Black locust breeding in Serbia 12.30 - 13.00 Discussion 13.00 - 14.30 Lunch 14.30 - 15.00 BREEDING08: Oral presentations by T. Warkentin and K. McPhee

15.00 - 16.00 Session 2: Legume breeding and biodiversity Chairs: Branko Ćupina and Petr Smýkal

15.00 - 15.30 Smýkal P.: Characterization of genetic diversity within pea (Pisum sativum L.) germplasm collections

15.30 - 15.45 Ćupina B., Mikić A., Mihailović V., Krstić Ð., Hauptvogel P.: Breeding potential of wild Vicia species

15.45 - 16.00 Discussion

16.00 - 16.15 Posters review and presentations of legume research publications Chairs: Anne-Marie Bochard and Aleksandar Mikić

16.15 - 16.30 Coffee break and poster exhibition

16.30 - 17.45 Session 3: Status and requirements of legume breeding Chairs: Lamiae Ghaouti and Fred Stoddard

16.00 - 16.30 Ghaouti L., Sass O.: Requirements of a practical legume breeding program (faba beans, peas) from the research community

16.30 - 16.45 Stoddard F.: New interest in grain legumes in Finland 16.45 - 17.00 Vasić M., Zdravković M.: Dry bean (Phaseolus vulgaris L.) breeding in Serbia

17.00 - 17.15 Radović J., Lugić Z., Sokolović D., Štrbanović R., Vasić T.: Breeding of white clover (Trifolium repens L.) and birdsfoot trefoil (Lotus corniculatus L.) in Serbia

17.15 - 17.30 Karagić Đ., Katić S., Mihailović V, Vasiljević S., Mikić A., Milić D.: What a seed producer needs from a plant breeder – The example of Novi Sad (NS) forage legume varieties

17.30 - 17.45 Discussion 19.30 Workshop dinner

Time FRIDAY, 28 NOVEMBER 2008

8.30 -10.30 GL-TTP General Assembly Chairs: Petr Smýkal and other members of GL-TTP Executive Committee

10.30 - 11.00 Coffee break

11.00 - 12.30 Session 4: Fundaments of legume breeding Chairs: Noel Ellis and Peter Winter

11.00 - 11.30 Winter P.: High-precision breeding: reliable and cost-efficient molecular tools are looking for users

11.30 - 11.45 Ubayasena L., Warkentin T., Bett K., Tar'an B., Bing D.: Genetic analysis, QTL mapping and gene expression analysis of key visual quality traits affecting the market value of field pea

11.45 - 12.00 Kumar Y., Mishra S. K., Tyagi M. C., Sarker A., Sharma B.: Linkage between genes for leaf size, leaf shape, stipule size, pod size and globe plant type in lentil (Lens culinaris Medik.)

12.00 - 12.15 Perić V., Drinić-Mladenović S., Srebrić M.: RAPD markers in estimating diversity of soybean parental genotypes and predicting traits of progeny

12.15 - 12.30 Discussion 13.00 - 14.30 Lunch

14.30 - 16.15 Session 5: Aims and methods of legume breeding Chairs: Tom Warkentin and Slobodan Katić

14.30 - 15.00 Vasiljević S., Lugić Z., Šurlan-Momirović G., Ivanović M.: Characteristics of special importance in red clover (Trifolium pratense L.) breeding

15.00 - 15.15 Gantner R.: Breeding value estimation of 6 pea genotypes based on diallel cross 15.15 - 15.30 Palmer R.: Soybean hybrids

15.30 - 15.45 Buhayov V. D., Maksimov A. M., Zadorozhnyi V. S.: Theoretical preconditions for the development of effective methods for selection of self-incompatible genotypes of Medicago sativa L.

15.45 - 16.00 Milić D., Katić S., Mikić A., Vasiljević S.: Genetic and phenotypic correlations among morphological and agronomic traits of lucerne genotypes

16.00 - 16.15 Discussion 16.30 - 17.00 Coffee break

17.00 - 18.30 Session 6: Legume breeding, symbiosis and stress Chairs: Alexey Borisov and Kevin McPhee

17.00 - 17.30

Borisov A., Ovchinnikova E., Nemankin T., Grishina O., Shtark O., Akhtemova G., Krasheninnikova A., Moloshonok A., Zhukov V., Kazakov A., Naumkina T., Vasilchikov A., Chebotar V., Gianinazzi-Pearson V., Tikhonovich I.: New insights in legume breeding: plant genetics of beneficial plant-microbe systems, evolution and applications in sustainable agriculture

17.30 - 17.45 Marinković J., Vasić M., Jarak M.: Effect of bean inoculation with Rhizobium leguminosarum bv. phaseoli on pod and grain number and grain mass

17.45 - 18.00 McPhee K., Wang X.: Genetic resistance to Phoma medicaginis in pea

18.00 - 18.15 Meglič V., Kavar T., Maras M., Kidrič M., Šuštar-Vozlič J.: The expression profiles of selected genes in leaves of different beans species (Phaseolus spp.) under drought stress

18.15 - 18.30 Discussion

18.30 - 19.00 Closing ceremony and farewell address Chairs: GL-TTP Executive Committee

INTEGRATING LEGUME SCIENCE AND CROP BREEDING

BOOK OF ABSTRACTS

SECOND GL-TTP WORKSHOP

NOVI SAD, SERBIA, 27 & 28 NOVEMBER 2008

INTEGRATING

LEGUME SCIENCE AND

CROP BREEDING

BOOK OF ABSTRACTS

SECOND

GRAIN LEGUMES

TECHNOLOGY TRANSFER PLATFORM

(GL-TTP)

WORKSHOP

MASTER CONGRESS CENTRE

NOVI SAD

SERBIA

27 & 28 NOVEMBER 2008

Second GL-TTP Workshop Book of Abstracts

Integrating Legume Science and Crop Breeding

Publishers:

Institute of Field and Vegetable Crops Maksima Gorkog 30

21000 Novi Sad Serbia

www.ifvcns.co.yu

Grain Legumes Technology Transfer Platform (GL-TTP) 38, rue de Berri

75008 Paris France

www.gl-ttp.com

Editors:

Paolo Annicchiarico Anne-Marie Bochard

Noel Ellis Aleksandar Mikić

Álvaro Ramos Monreal Petr Smýkal

Tom Warkentin Peter Winter

Cover design:

Aleksandar Mikić Legumes: the Bright Side of the World

in memory of

Richard Wright (1943-2008) and Aleksandar Popović (1973-1999)

Programme Committee

Paolo Annicchiarico (Centro di Ricerca per le Produzioni Foraggere e Lattiero-Casearie, Lodi, Italy)

Anne-Marie Bochard (Limagrain, Riom, France)

Noel Ellis (John Innes Centre, Norwich, United Kingdom)

Aleksandar Mikić (Institute of Field and Vegetable Crops, Novi Sad, Serbia)

Álvaro Ramos Monreal (Consejería de Agricultura y Ganadería and Universidad de Valladolid, Valladolid, Spain)

Petr Smýkal (Agritec Plant Research Ltd., Šumperk, Czech Republic)

Tom Warkentin (University of Saskatchewan, Saskatoon, Canada)

Peter Winter (GenXPro GmbH, Frankfurt, Germany)

Local Organising Committee

Branko Ćupina

(Faculty of Agriculture, Novi Sad, Serbia) Borislav Kobiljski

(Institute of Field and Vegetable Crops, Novi Sad, Serbia) Vojislav Mihailović

(Institute of Field and Vegetable Crops, Novi Sad, Serbia) Aleksandar Mikić

(Institute of Field and Vegetable Crops, Novi Sad, Serbia) Dragana Miladinović

(Institute of Field and Vegetable Crops, Novi Sad, Serbia) Jegor Miladinović

(Institute of Field and Vegetable Crops, Novi Sad, Serbia) Nevena Nagl

(Institute of Field and Vegetable Crops, Novi Sad, Serbia) Mirjana Vasić

(Institute of Field and Vegetable Crops, Novi Sad, Serbia) Sanja Vasiljević

(Institute of Field and Vegetable Crops, Novi Sad, Serbia)

Foreword On behalf of GL-TTP Executive and the Workshop Organising Committees, I am very pleased to welcome you at the 2nd GL-TTP Workshop Integrating Legume Science and Crop Breeding, held on 27 and 28 November 2008, at Novi Sad, Serbia. GL-TTP (Grain Legumes Technology Transfer Platform) has been established as a not-for-profit organisation to bridge the gap between research-oriented community and breeders focused on variety improvement, both aimed at production and quality improvements of grain legumes. GL-TTP was established in 2005 within the frame of EU funded Grain Legumes Integrated Project (GLIP) (2004-2008) to ensure the exploitation of the project outputs by the grain legume community. Having thus a basis both in fundamental research and industry-based world, GL-TTP is in good position to identify the specific needs and constraints of grain legume breeders and growers and to channel the latest research results and technologies to the grain legume industry. The structure was launched on AEP initiative through the means and activities of GRAIN LEGUMES Integrated Project which ended in February 2008, GL-TTP is now standing on its own foot and resources for a year, a situation which is difficult because of the current decreasing means and interests devoted to legumes in the difficult economic context of European breeders and growers. We have thus witnessed exodus of first GL-TTP members, joined now by new, “second generation” GL-TTP members, often from non-EU countries including Eastern and Southern Europe. We hope this new trends prove viable and provides novel drive into GL-TTP activities. Although world-wide oriented, GL-TTP was primarily set up and based in Europe: the activities have been mainly focussed on temperate species of grain legumes adapted to European countries as well as Canada and Australia, and have taken European context largely into account. Today grain legumes are grown on less than 3% of arable land in Europe, in spite of their key role of protein temperate plant source and of their benefits as vital component of crop rotation system (thanks their symbiotic nitrogen fixation and their break crop effects). Situation might hopefully change with the increasing environmental concerns, the current high fertilizers costs and further strengthened demand in protein sources for industry. Although, GL-TTP and GLIP activities focused till know dominantly on temperate region grain legumes: as pea, faba bean, lentil, chickpea, vetches, lupine, we would like to make GL-TTP attractive to entire community working and using any species of large Fabaceae family, the third largest plant family on the Earth. Thus we would like to address also important sector of forage legumes, including clover and alfalfa, legume trees as fast growing Robinia and Acacia species and ultimately also soybean breeders and producers, traditionally viewed as competitors. Although soybean benefit from specific status of oil crop, physiologically and phylogenetically it clearly belongs to legumes. Moreover, most of the genomic tools are becoming to benefit from knowledge of two model legumes (Medicago truncatula and Lotus japonicus) genomes and can be already directly and successfully applied to most legume species. As plant breeding is traditionally focussed on identification and subsequent combination of selected parental genotypes, there are challenges to assist this timely and often still intuitive process by molecular tools in order to shorten and make it more efficient, the process of desired combination identification. Such process inevitably starts at germplasm collections, which should

safeguard the genetic richness of species. It is widely recognized recognised that the genetic diversity of cultivated plants has narrowed as a result of thousands of years of domestication and connected bottlenecks. With the advent of not only traditional microsatellite or other repetitive sequences based, but gene- specific markers, we become to discover the hidden genetic variability. More then ever before, we are aware of the danger of diversity loss, linked to cultivation of limited number of high-yielding varieties. Moreover we are much more aware of the benefits resulting from the exploitation of older varieties, landraces and even wild crop relatives, for breeding new varieties to cope with environmental and demographic changes. Most of these above mentioned issues we would like to address during the two days meeting, aim to enhance communication and interaction among legume community. To facility this purpose, the workshop has been set up in interactive sessions, with most of the participants being able to have talks. We would like to initiate throughout the workshop, set up of research and development and technology transfer projects and to discuss the strategies for getting such projects funded. We wish this 2nd GL-TTP Workshop to be a success and wish you all to enjoy it! Last but not least we would like to express our gratitude to local organisers, without which enthusiasm and effort this meeting would not be possible. Petr Smykal, GL-TTP President, with GL-TTP Executive Committee members, Anne-Marie Bochard, Treasurer, Tom Warkentin, First Vice-President, Aleksandar Mikić, Second Vice-President, Noel Ellis, Past President, Catherine Golstein, Past Scientific Manager, Anne Schneider, Executive Secretary

Contents 13

Session 1: Achievements and challenges in legume breeding

14 T. H. N. Ellis: Genetics and breeding in legumes

16 B. Sharma: Present status of genetic research in lentil (Lens culinaris)

18 V. Đorđević, J. Miladinović, S. Balešević-Tubić and V. Đukić: Future perspective in soybean

breeding 20

P. Annicchiarico and C. Scotti: Some burning questions for lucerne breeders 22

B. Kovačević, S. Orlović and A. Pilipović: Black locust breeding in Serbia 24

M. Imtiaz: Breeder’s needs – conventional versus molecular breeding in grain legumes 27

Session 2: Legume breeding and biodiversity

28 P. Smýkal: Characterization of genetic diversity within pea (Pisum sativum L.) germplasm

collections 30

S. Angelova: Maintenance, enrichment and utilization of grain legume collections in Bulgaria .32

B. Ćupina, A. Mikić, Đ. Krstić, V. Mihailović and P. Hauptvogel: Breeding potential of wild Vicia species

34 A. Mikić, V. Mihailović, B. Ćupina, D. Jovićević, D. Milić and Đ. Krstić: Evaluation of seed yield

components in red-yellow pea (Pisum fulvum Sm.) 36

B. Ćupina, V. Mihailović, A. Mikić, P. Erić and Đ. Krstić: Evaluation of seed yield components in Ethiopian pea (Pisum sativum subsp. abyssinicum (A. Braun) Govorov)

38 J. Rysová, J. Ouhrabková, D. Gabrovská, I. Paulíčková and T. Vymyslický: The possibilities of

grasspea utilization in foods 40

Đ. Krstić, B. Ćupina, V. Mihailović, A. Mikić and R. Hauptvogel: Potential for forage and grain yields in Lathyrus species

42 M. Ljuština and A. Mikić: Grain legumes technology transfer in Old Europe - archaeological

evidence 44

A. Mikić, M. Ljuština, V. Đorđević, B. Ćupina, V. Mihailović and S. Vasiljević: Grain legumes technology transfer in Old Europe - linguistic evidence

47 Session 3: Status and requirements of legume breeding

48 L. Ghaouti and O. Sass: Requirements of a practical legume breeding program (faba beans, peas)

from the research community 50

F. L. Stoddard: New interest in grain legumes in Finland 52

M. Vasić and M. Zdravković: Dry bean (Phaseolus vulgaris l.) breeding in Serbia 54

J. Radović, Z. Lugić, D. Sokolović, R. Štrbanović and T. Vasić: Breeding of white clover (Trifolium repens L.) and birdsfoot trefoil (Lotus corniculatus L.) in Serbia

56 Đ. Karagić, S. Katić, V. Mihailović, S. Vasiljević, A. Mikić and D. Milić: What a seed producer

needs from a plant breeder – the example of Novi Sad (NS) forage legume varieties 58

R. Gantner, M. Stjepanović, S. Popović and T. Čupić: Breeding pea for grain and fodder in Osijek

60 N. Sarukhanyan and A. Vanyan: World chickpea collection variety testing in mountainous

regions of the Republic of Armenia 62

V. F. Petrychenko, S. V. Ivanyuk and S. I. Kolisnyk: Theoretical foundations and practical results of adaptive soybean selection in Ukraine

64 R. Darai, B. N. Adhikari, R. K. Neupane, B. S. Bastola and B. R. Baral: Participatory Plant

Breeding (PPB) approach for evolving superior varieties of soybean (Glycine max L. Merril) under maize based system in Nepal

67

Session 4: Fundaments of legume breeding

68 P. Winter: High-precision breeding: reliable and cost-efficient molecular tools are looking for

users 70

L. Ubayasena, T. Warkentin, K. Bett, B. Tar'an and D. Bing: Genetic analysis, QTL mapping and gene expression analysis of key visual quality traits affecting the market value of field pea

72 Y. Kumar, S. K. Mishra, M. C. Tyagi, A. Sarker and B. Sharma: Linkage between genes for leaf

size, leaf shape, stipule size, pod size and globe plant type in lentil (Lens culinaris Medik.) 74

V. Perić, S. Mladenović-Drinić and M. Srebrić: RAPD markers in estimating diversity of soybean parental genotypes and predicting traits of progeny

76 A. I. Adesoye and A. Emese: In vitro regeneration of African yam bean (Sphenostylis stenocarpa

(Hochst et A. Rich) Harms.) by direct organogenesis 78

D. Steinhauer, F. Khan, N. Fatnassi, C. Molina, R. Horres, C. Abdelly, M.-J. Delgado, G. Kahl and P. Winter: Quantitative Real-Time-PCR: fast and efficient technology for knowledge-based

breeding

80 P. Smýkal: From lab to field: the case study of molecular tools for marker assisted breeding in

pea virus and fungal resistances 82

K. Taški-Ajduković, V. Ðorđević, M. Vidić, J. Miladinović and M. Vujaković: Differences in seed proteins among high-protein soybean genotypes

85

Session 5: Aims and methods of legume breeding

86 S. Vasiljević, Z. Lugić, G. Šurlan-Momirović and M. Ivanović: Characteristics of special

importance in red clover (Trifolium pratense L.) breeding 88

R. Gantner, G. Drezner, M. Stjepanović, S. Popović and T. Čupić: Combining ability of pea genotypes

90 R. Darai1, A. Sarker2, N. K. Yadav3, D. B. Gharti3, B. N. Adhikari3 and D. N. Pokhrel: Farmer-

participatory selection and scaling up lentil varieties in Nepal 92

M. Srebrić and V. Perić: Conventional breeding of Kunitz-free soybean genotypes 94

V. D. Bugayov, A. M. Maksimov and V. S. Zadorozhnyi: Theoretical preconditions for the development of effective methods for selection of self-incompatible genotypes of Medicago sativa

96 D. Milić, S. Katić, A. Mikić and S. Vasiljević: Genetic and phenotypic correlations among

morphological and agronomic traits of lucerne genotypes 98

V. Mihailović, A. Mikić, S. Katić, Đ. Karagić and I. Pataki: Grain yield components in afila (af) lines of field pea (Pisum sativum L.)

100 R. Matić1, S. Nagel1, G. Kirby1, I. Young2 and K. Smith: Challenges in breeding vetches (Vicia

spp.) for diverse purposes 102

M. E. Tate, D. Chowdhury and H. Firincioglu: In search of a low toxin feed vetch 104

V. Đorđević, J. Miladinović, S. Balešević-Tubić, V. Đukić and A. Mikić: Bulk segregation analysis and improvement of seed quality in soybean

106 M. M. Abdel-Galil and N. M. Hamed: Evaluation of yield potential, genetic variances and

correlation for nine cultivars of alfalfa under the New Valley environment 108

M. M. Abdel-Galil, A. A. Helmy and N. M. Hamed: Developing a synthetic population through selection in Egyptian clover genotypes (Trifolium alexandrinum L.)

111

Session 6: Legume breeding, symbiosis and stress

112 A. Borisov, E. Ovchinnikova, T. Nemankin, O. Grishina, O. Shtark, G. Akhtemova, A.

Krasheninnikova, A. Moloshonok, V. Zhukov, A. Kazakov, T. Naumkina, A. Vasilchikov, V. Chebotar, V. Gianinazzi-Pearson and I. Tikhonovich: New insights in legume breeding: plant

genetics of beneficial plant-microbe systems, evolution and applications in sustainable agriculture

114 J. Marinković, M. Vasić and M. Jarak: Effect of bean inoculation with Rhizobium leguminosarum bv.

phaseoli on pod and grain number and grain mass 116

K. McPhee and X. Wang: Genetic resistance to Phoma medicaginis in pea 118

V. Meglič, T. Kavar, M. Maras, M. Kidrič and J. Šuštar-Vozlič: The expression profiles of selected genes in leaves of different beans species (Phaseolus spp.) under drought stress

120 P. M. Gresshoff, A. Miyahara, S. Nontachaiyapoom, A. Indrasumunar, Y.-H. Lin, D. Reid, M.

Kinkema, B. Biswas, Q. Jiang, M. Miyagi, D. Li, M.-H. Lin, B. Carroll, P. K. Chan, T. Hirani, A. Kereszt and B. Ferguson: Systemic and local regulation of legume nodulation

122 D. Delić, O. Stajković, Đ. Kuzmanović, N. Rasulić, S. Maksimović and B. Miličić: Nitrogen

fixation efficiency of several rhizobial strains in symbiosis with Vigna angularis (L.) Willd., Vigna radiata (L.) Wilczek and Vigna mungo (L.) Hepper under field conditions

124 A. Korakhashvili: Legumes seed inoculation advance technology for biological nitrogen fixation

improvement 126

D. Rubiales, M. Fernández-Aparicio, A. Moral, A. Pérez-de-Luque, E. Barilli, J. C. Sillero, A. M. Torres, M. Curto, M. A. Castillejo, E. Prats and S. Fondevilla: Progress in pea breeding for

disease resistance at Córdoba, Spain 128

D. Petrović, M. Ignjatov, F. Bagi, M. Milošević, M. Vasić and M. Vujaković: Incidence of viruses in the most important bean varieties in Vojvodina

130 M. Ignjatov, M. Vidić, M. Milošević, J. Balaž, M. Vujaković, D. Petrović: Race identification of

Pseudomonas syringae pv. glycinea on commercial soybean varieties in Serbia 132

N. A. Roder, T. Y. Bogracheva, D. A. Jones and C. L. Hedley: Pod growth temperature affects the granular structure of starch from pea (Pisum sativum L.) seeds

134 M. Vasić, A. Mikić, V. Mihailović, B. Ćupina, Đ. Krstić and G. Duc: Grain yield components in

winter genotypes of faba bean (Vicia faba L.) 136

V. Mihailović, A. Mikić, B. Ćupina and M. Vasić: Challenges in breeding subtropical legumes for temperate regions

138 M. Vishnyakova and A. Mikić: White lupin (Lupinus albus L.) landraces and the breeding for

tolerance to alkaline soil reaction 141

Index of authors 143

List of participants


Second GL-TTP Workshop Integrating Legume Science and Crop Breeding, Novi Sad, Serbia, 27 & 28 November 2008

Genetics and breeding in legumes

T. H. N. Ellis

Department of Crop Genetics, John Innes Centre, Colney Lane, Norwich, NR4 7UH [email protected] Legume genetics is a rich and venerable subject: we have only to recall the names Mendel and Vavilov to understand this. However, past glories are past and we need to ask what legume genetics can do now and how it can impact on breeding. In this presentation I will discuss the usefulness of comparative genomics tools, the potential for even higher resolution genotyping and the availability of novel genetic resources. In this I will draw heavily on the outcomes of the Grain Legumes Integrated Project. Despite the optimism and eagerness we can have for legume genetics and genomics, I will temper this by addressing the issue of how such advances can influence the three central issues of breeding: heritability, selection intensity and phenotypic variance.

14


Notes

15


Present status of genetic research in lentil (Lens culinaris)

B. Sharma

Division of Genetics, Indian Agricultural Research Institute, Delhi 110012, India [email protected] The genetic research in lentil has a short history of about three decades. More than a dozen short mapping sequences have been identified with the help of molecular markers, the seven chromosomes still do not have their individual linkage maps. Nevertheless, each of the chromosomes can be distinguished by a sequence of molecular markers associated with them. A major bottleneck in genetic analysis of lentil is non-availability of markers for easily identifiable functional genes with visible phenotype or isozyme alleles at different loci. A collection of a few hundred visible, easily detectable and quantifiable markers is a prerequisite for a meaningful mapping analysis. Experience has shown that not more than about three dozen traits with monogenic inheritance are available in the natural germplasm of lentil. Almost an equal number of enzyme markers have also been discovered. Induced mutations are the only means to discover new genes. Mutations have been reported for many morphological characters, such as, chlorophyll deficiency, plant height, growth habit, branching pattern, stem structure, leaf morphology, inflorescence, calyx shape, flower structure, pod characters, sterility, seed coat colour, curled apex, radicle hypertrophy, cotyledon leaves, tricotyly, folded leaflets, globular plant structure, compact, waxless, acute pod, tendril structure, stem fasciation, etc. Genes have been identified in germplasm collections for growth habit (Ert), red stem (Gs), red pod (Rdp), brown leaf (Bl), light green foliage (Gl), plant height (Ph), pubescence (Pub), high leaflet number (Hl), oval–acute leaf (Ol), broad leaflet (Blf), stipule size (Lst), large pod (pd), globe mutation (Glo), tendril formation (Tnl), stem fasciation (Fas), flower colour (V, P), testa spotting (Mot, Spt), brown testa (Brt), flower number (Fln), pod dehiscence (Pi), testa colour (Blt, Ggc, Tgc), seed coat pattern (Scp), tannin in seed coat (Tan), hard seed (Hsc), cotyledon colour (Y, B, Dg), Fusarium wilt tolerance (Fw), frost tolerance (Rf, Frt), early flowering (Sn), anthracnose tolerance (Lct). Donors have been identified for several economic traits. These include winterhardiness, drought tolerance, soil factors (salinity, boron excess and deficiency, iron), herbicide resistance, and resistance to rust, wilt, blight, anhracnose, stemphylium, powdery mildew, and viral diseases. Many of these genes have been mapped in linkage studies and others tagged with molecular markers. Allozyme variations have also been used extensively to create more than a dozen short linkage sequences. However, only four functional genes have been included in molecular mapping. Quantitative trait loci (QTLs) have been mapped for height of first branching (5), plant height (3), flowering time (5), pod dehiscence (7), and winter survival (9).

16


Notes

17


Future perspective in soybean breeding

V. Đorđević, J. Miladinović, S. Balešević-Tubić and V. Đukić

Institute of Field and Vegetable Crops, Novi Sad, Serbia [email protected] Plant breeding usually considers selecting superior genotype from large and variable plant populations. Widespread use of molecular markers in study of complex, quantitative traits has inadequate application in plant breeding. Reasons for this disproportion lying in the nature of complex traits. Main concepts of application molecular markers in soybean breeding consider QTL detection, developing procedure for marker based selection and estimates of QTL effect (1). Success of appropriate molecular marker application concept possesses several challenges: type of molecular markers, quality of phenotypic data, epitasis, G x E interaction, QTL x environment and QTL x genetic background interaction. Soybean genome is covered by 4000 different types of markers, it is precondition for successful maping of complex traits and marker assisted selection (2). Due to inconsistency of QTLs, simple transfer of QTLs in to new soybean variety is very difficult and uncertain. Even closely related bi parental populations, whit one common genotype, have different QTLs for complex traits (3). Accordingly to this fact, breeding procedure need to involve QTL analysis whit simultaneously selection of superior progeny. One time embedded linkage between molecular markers and QTL can be used for screening seed remnant of that bi parental population. (1) Bernardo R. (2008) Crop Science, 48, 1649-1664. (2) Choi I., Hyten D., Matukumalli L., Song Q., Chaky J., Quigley C., Chase K., Lark G., Reiter R., Yoon M., Hwang E., Yi S., Young N., Shoemaker R., Tassell C., Specht J. and Cregan P. (2007) Genetics, 176, 685–696. (3) Orf J., Chase K., Jarvik T., Mansur L., Cregan P., Adler F. and Lark K. (1999) Crop Science, 39, 1642–1651.

18


Notes

19


Some burning questions for lucerne breeders

P. Annicchiarico and C. Scotti

CRA - Centro di Ricerca per le Produzioni Foraggere e Lattiero-Casearie, viale Piacenza 29, 26900 Lodi, Italy [email protected] Lucerne has displayed a low rate of genetic gain for forage yield compared with other species (e.g. maize), owing to several biological characteristics which hinder the breeding work (autotetraploidy; open pollination; high gene interaction; perennial growth cycle; high genotype × environment interaction). Without any claim to be exhaustive, we point out some burning questions which we feel as crucial in our selection work. a) Which is the most cost-efficient selection scheme? A number of schemes have been proposed for lucerne to cope with its characteristics, but empirical comparisons have been rare and limited to just a few schemes whereas theoretical comparisons are hindered by the absence of reliable estimation for relevant genetic parameters. A large comparison of selection schemes is on-going in Lodi. b) How exploiting heterosis? Non additive genetic variation, which is at the basis of heterosis, could be exploited by free hybrids (alias semi-hybrids), in which both hybrid seed (derived from interpopulation crossing) and non-hybrid seed (derived from intrapopulation crossing) are present. Two procedures implying a different level of complexity are under testing in Lodi. The production of hybrid varieties (75% hybridity) through a patented male-sterility technique is gaining importance, especially in US. c) Can we breed very drought-tolerant varieties? This issue, of increasing interest in several regions as a consequence of climate change, has been little investigated in terms of genetic variation and underlying physiological mechanisms. Information for Mediterranean environments is being generated by the EU-funded project PERMED. d) How to improve forage quality? Early mowing has a major impact on forage quality, but requires varieties which tolerate frequent cutting. Selection for modified stem morphology (increased internode number, decreased internode length) has good potential for improving intrinsic forage quality (e.g. protein yield) based on our results, which also suggest some opportunities for the conventional simultaneous improvement of forage yield and leaf/stem ratio. e) Which opportunities for marker-assisted selection? Various linkage maps for tetraploid lucerne are available now, but results from US trials suggest that useful markers for forage yield are markedly site-specific and little consistent across cropping years. Methods based on the analysis of shifts in marker allele frequency in selected material relative to unselected one seem useful in breeding for stress tolerance. There is modest evidence so far on the relationship between marker-based genetic diversity and heterosis of parent material for synthetic varieties. f) How to exploit genetic tools from M. truncatula in lucerne breeding? Markers currently available for lucerne have essentially been developed for M. truncatula. This model species can be very useful also for identifying genes which control metabolic pathways of agronomic interest – also active in lucerne and usable in gene-based marker-assisted selection, as described for saponin content and composition. Information on marker-trait associations for forage yield or tolerance to abiotic stresses obtained from M. truncatula may be little exploitable in M. sativa, owing to the physiological differences arising from perenniality besides the possible differences due to gene function or gene structure in the genome.

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Black locust breeding in Serbia

B. Kovačević, S. Orlović and A. Pilipović

Institute for Lowland Forestry and Environment, Antona Čehova 13, 21000 Novi Sad, Serbia branek@uns. ns.ac.yu Black locust (Robinia pseudoacacia L.) is species originated from North America. It was introduced in Europe by Robin in 1601, and spread very fast through the continent. The interest in this species was related to its high adaptability, drought tolerance, abundant and frequent seed crop, excellent sprouting ability, early and abundant flowering, fast growth and relatively high timber yield, tolerance to pests and diseases. It is mainly used for fixing sands, afforestation of abandoned agricultural lands, wood production for industry and energy and honey production. The breeding efforts are concentrated in improvement of tree shape (straight), wood quality (for industrial purpose), long flowering and high nectar production. In Serbia the work on the improvement of black locus is focused on the establishment of seed orchards established from quality stands or with superior clones gained by means of vegetative propagation (root cuttings, green cuttings and micropropagation). The variability of important anatomical, physiological and morphological traits is studied (2, 3). Also, the genotypes with desired properties are preserved in form of genotype collections designated to the production of vegetative propagation material. The establishment of commercial plating material production by means of vegetative propagation is only planed for clones of ornamental and apicultural significance (1). The contemporary research in world goes for identification of biochemical and molecular markers that are important for variability studies and protection of authorship, as well as identification of quantitative trait loci that could be used in marker assisted selection.

(1) Kevrešan S., Kovačević B., Ćirin-Novta V., Kuhajda K., Kandrač J., Pavlović K. and Grbović Lj. (2007) Journal of the Serbian Chemical Society, 72, 953-959. (2) Orlović S., Pajević S., Krstić B., Merkulov Lj., Nikolić N. and Pilipović A. (2004) Matica Srpska Proceedings of Natural Sciences, 106, 65-79. (3) Tomović Z., Orlović S., Guzina V., Ivanišević P. (1997) Genetika, 29, 1, 23-30.

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Breeder’s needs – conventional versus molecular breeding

in grain legumes

M. Imtiaz International Center for Agricultural Research in the Dry Areas (ICARDA), P.O. Box 5466, Aleppo, Syria [email protected] Conventional plant breeding (CPB) exploits natural environments to breed new varieties of crops to suit different agro ecological conditions, cope with biotic or abiotic stresses better, improve nutritional value, and use water or nutrients more efficiently. Besides tremendous achievements in crop improvement through phenotypic selection, significant difficulties are often encountered in phenotypic selection, such as genotype by environment interaction, and expensive, unreliable and time consuming screening methodologies for target traits. With the advent of molecular biology, the term molecular breeding (MB) has been introduced which is the application of genomics tools such as molecular markers to facilitate CPB at critical stages of varietal development. Cereals breeders have changed their focus for difficult traits such as disease resistance, aluminum/boron tolerance from phenotypic selection to genotypic selection in order to increase the certainty of successful outcomes while for grain legumes breeders particularly chickpea, lentil and faba bean, MB is still in its infancy stage. Grain legumes breeders desire to use MB to significantly reduce time to develop a new variety, have more effective and direct control of the alleles retained and discarded, access to new genes that may provide greater diversity, and avoid genotype-environment interaction. For example the International chickpea improvement program at the International Centre for Agricultural Research in the Dry Areas (ICARDA) focuses on ascochyta blight (AB), Fusarium wilt (FW), leaf miner (LM), and cyst nematode (CCN) resistances while drought and cold tolerance are the breeding targets for abiotic stresses. To select AB resistant germplasm through CPB, the strategy includes planting of 6 hectare disease nursery comprising 16000 to 18000 lines under mist irrigation each year. Despite artificial inoculation and providing all conductive environments, the diseases development is not sufficient in some years to provide a good screen to select for AB resistance. This leads to the increase in the number of lines to be screened in subsequent planting season which require more resources in terms of area to be planted and constraints on financial resources. A gene-based or perfect marker for AB could help to make reliable selection at seedling stage without relying on disease development in the field. Similarly, there is a need for tightly linked molecular markers for other traits to be developed and deployed in the breeding programs which can help grain legume breeders to combine desirable alleles at a greater number of loci and at earlier generations than is possible with CPB methodologies. For the grain legume industry to integrate new technologies into CPB efforts, there is strong need to 1) link the genomics resources and theoretic knowledge available with application in practical breeding through consultation among molecular biologist, pre-breeder, and breeders to define breeding objectives 2) establish a common forum for legumes scientists such as website to have free access to genomics and other genetic resources 3) develop understanding among scientists in which the ultimate goal is to produce better varieties faster which are acceptable to farmers and other end users. (1) Bonnett D. G., Rebetzke G. J. and Spielmeyer W. (2005) Mol. Breeding, 15, 75-85. (2) Babu R., Nair S. K., Prasanna B. M. and Gupta H. S. (2002) Current Sci., 87, 607-619.

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Characterization of genetic diversity within pea (Pisum

sativum L.) germplasm collections

P. Smýkal

Agritec Plant Research Ltd., Plant Biotechnology Department, Šumperk, Czech Republic [email protected] Several large germplasm collections of the Pisum genus are conserved and maintained world-wide. To facilitate the management and increase efficiency of the use, a core collection is being currently developed. Also for breeding it is important to know the genetic basis of cultivars, especially to see if it has not become too narrow to render crops more vulnerable to diseases or pests. Additionally accessions which display DNA profiles most distinct from others are likely to contain the greatest number of novel alleles, which can be exploited in breeding. Thus, traditionally descriptions were made by morphological descriptors in several growth seasons owing to variation and environmental influence, together with known pedigree and passport data. Moreover, over last years genetic structure has been investigated by several molecular markers, including microsatellite SSR and retrotransposon-based markers. The emphasis was put on co-dominant, RBIP (Retrotransposon-Based Insertion Polymorphism) markers based on the PDR-1 element (Jing et al. 2005) and microsatellite loci. Additionally, high copy elements, were successfully applied in multiplex IRAP-PCR (Inter-Retrotransposon Amplified Polymorphism) format (Smýkal 2006, Smýkal et al. 2008) suitable for fast variety (Zong et al. 2008) fingerprinting. With all these markers, several major world pea germplasm have been analyzed and core collections formed. Thus over 2,000 accessions including 1,200 pea accessions of Chinese origin from Australian Temperate Field Crops Collection, were analyzed by 21 SSR loci (Zong et al. 2008), 504 of nearly 4,000 USDA pea germplasm have been assessed by 37 RAPD and 15 SSR markers (Coyne et al 2005, Brown et al. 2007), INRA France used extensive set of SSR markers to genotype (Burstin et al. 2001, Baranger et al. 2004, Loridon et al. 2005), CDC Canada pea collection was studied by RAPD, ISSR and SSR, JIC pea germplasm composed in large part by expedition collections was analyzed by over 65 RBIP markers (Flavell et al. personal communication) and we have genotyped over 1,200 pea accessions held at Czech National Pea Germplasm by combination of RBIP and SSRs (Smýkal et al. 2008). All together, large amount of scorable polymorphic data points have been produced for over 7,000 accession, which were subsequently subjected to genetic distance analysis and/or model-based Bayesian method. However after data processing, further use of such data is limited, without permitting important cross-comparison. Furthermore, most of these accessions have been evaluated for morphological, agronomical and phytopathological traits, the data of enormous scientific and breeding potential. With advance of model legume sequencing and our genomic knowledge, we are witnessing switch to gene-based markers (Jing et al. 2007). Those together with above mentioned might be used for selected QTL and modern association mapping (Zhu et al. 2008), as applied already for more exploited cereals. The are several key points of such large but variable data further exploitation, selection and use of common set of markers applied to developed core sets together with deposition and availability of both molecular and agronomical data, creating virtual world-wide pea germplasm for our common benefit. Acknowledgements: This work was financially supported by Ministry of Education of Czech Republic research project MSM 2678424601 and INGO LA08011.

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Maintenance, enrichment and utilization of grain legume

collections in Bulgaria

S. Angelova

Institute of Plant Genetic Resources, Sadovo, Bulgaria [email protected] The grain legume collections are the richest group in plant world, with their economic value and dissemination they take second place after cereals. The Bulgarian grain legume collections include diverse genetic material, belongs to nine genera. Studied and preserved mainly at the IPGR in Sadovo. The three larger collections of grain legumes – peas, dry beans and vetch are represented by the following more important species: Pisum sativum and Pisum arvense, Ph. vulgaris and Vicia sativa. The Bulgarian accessions are mostly old local cultivars and populations, newly bred and cultivated varieties, breeding lines etc. Breeding for winter hardiness is being made for pea, vetch, lentil and faba beans. Dominant for all collections are the selected varieties. Special attention is given to old local varieties, populations and wild relatives (peas, dry beans, vetch and cowpea). The important quality characters studied in grain legume crops are connected directly with breeding trends: earliness, drought tolerance, plant productivity, and protein content. The main purpose of this study is to analyse status of the grain-legumes collections at the IPGR and to systematize the most important characteristics for breeding and agriculture. The grouping of accessions on the most valuable characters gives possibilities for creating of new varieties with high potential of yield and quality of production.

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Breeding potential of wild Vicia species

B. Ćupina1, A. Mikić2, Đ. Krstić1, V. Mihailović2 and P. Hauptvogel3

1University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia 2Institute of Field and Vegetable Crops, Novi Sad, Serbia 3Research Institute of Plant Production, Piešťany, Slovakia [email protected] Vetches are largely present in the flora of Serbia and are regarded as one of the oldest forage crops in the country. The Annual Forage Legumes Collection (AFLCNS) of the Institute of Field and Vegetable Crops in Novi Sad is constantly enriched with the wild populations of Serbian origin, collected mostly in the region of Novi Sad and the mountain of Fruška Gora (2). By the end of 2008, AFLCNS had 1,276 accessions of 22 vetch species. The majority of the collected accessions belong to large-flowered vetch (Vicia grandiflora Scop.), narrow-leafed vetch (Vicia sativa L. subsp. nigra (L.) Ehrh.) and hairy vetch, with all three extremely widespread not only in the countryside, but also in urban areas, such as Belgrade or Novi Sad (1). In a long-term evaluation of the most important agronomic characteristics, the wild populations of majority of vetch species, especially of large-flowered, narrow-leafed and hairy vetch, have shown a high level of variability of the characteristics of agronomic importance related to forage and may be regarded as having a considerable potential for the development of the first Serbian cultivars of these species, suitable for forage production and green manure. Although the main obstacles of the breeding programme with wild vetches populations remain indeterminate stem growth, non-uniform maturity, pod dehiscence and large proportion of hard seeds, it may be expected that genotypes with desirable characteristics are able to be found, isolated and used in hybridisation. (1) Ćupina B., Krstić Đ., Mihailović V., Mikić A. and Vasiljević S. (2007) Bioversity International Newsletter for Europe, 34, 12. (2) Tomić Z., Đukić D., Katić S., Vasiljević S., Mikić A., Milić D., Lugić Z., Radović J., Sokolović D. and Stanisavljević R. (2005) Acta Agriculturae Serbica, X, 19, 3-16.

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Evaluation of seed yield components in red-yellow pea

(Pisum fulvum Sm.)

A. Mikić1, V. Mihailović1, B. Ćupina2, D. Jovićević1, D. Milić1 and Đ. Krstić2

1Institute of Field and Vegetable Crops, Novi Sad, Serbia 2University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia [email protected] According to the majority of modern botanical classifications, red-yellow, tawny or, simply, wild pea (Pisum fulvum Sm.) is regarded as another of the two species in the genus Pisum L., along with P. sativum L., common pea (2). Recently, this species is generally considered a source of the resistance to pea weevil (Bruchus pisorum L.), the most important pest in pea in many regions with arid climate (1). Within the Annual Forage Legumes Collection (AFLCNS) of the Institute of Field and Vegetable Crops in Novi Sad, there are 15 accessions of red-yellow pea of diverse geographic origin. All of them have been included in the evaluation of the most important agronomic characteristics, mostly because of the need for a better understanding of the relationship between yield components in this species and its hybrid progenies with common pea. A small-plot trial was carried out on a chernozem soil at the Experiment Field of the Institute of Field and Vegetable Crops at Rimski Šančevi during 2005, 2006 and 2007. The trial included twelve red-yellow pea accessions from AFLCNS. There were monitored plant height (cm), first fertile node height (cm), number of nodes (plant-1), ordinal number of the first fertile node, number of stems (plant-1), number of pods (plant-1), number of seeds (plant-1), thousand seeds mass (g), seed yield (g plant-1), harvest index and plant mass (g). The highest seed yield was in the accession 868277 (2.44 g plant-1). Seed yield per plant was in the highest positive simple correlations with number of seeds per plant (r = 0.937) and number of pods per plant (r = 0.882). (1) Byrne O. M. T. (2005) PhD Thesis. University of Western Australia, Perth, Australia, 135. (2) Mihailović V., Mikić A. and Ćupina B. (2004) Acta Agriculturae Serbica, IX, 17 (special issue), 61-65.

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Evaluation of seed yield components in Ethiopian pea (Pisum sativum subsp. abyssinicum (A. Braun) Govorov)

B. Ćupina1, V. Mihailović2, A. Mikić2, P. Erić1 and Đ. Krstić1

1University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia 2Institute of Field and Vegetable Crops, Novi Sad, Serbia [email protected] The majority of modern botanical classifications classify Ethiopian pea within the species Pisum saivum L. and gives it the rank of subspecies, that is, Pisum sativum subsp. abyssinicum (A. Braun) Govorov. On the other hand, some research reveals a possibility that it may be considered a separate species (1), that is, the third within the genus (3). Ethiopian pea is, in a similar way to other wild pea taxa, often a source to important diseases (2). Within the Annual Forage Legumes Collection (AFLCNS) of the Institute of Field and Vegetable Crops in Novi Sad, there are 15 accessions of Ethiopian pea of diverse geographic origin. All of them have been included in the evaluation of the most important agronomic characteristics, in order to get a better comprehension of the relationship between forage and seed yield components in both this species and its hybrid progenies with common pea and its subtaxa. A small-plot trial was carried out on a chernozem soil at the Experiment Field of the Institute of Field and Vegetable Crops at Rimski Šančevi during 2005, 2006 and 2007. The trial included twelve red-yellow pea accessions from AFLCNS. There were monitored plant height (cm), first fertile node height (cm), number of nodes (plant-1), ordinal number of the first fertile node, number of stems (plant-1), number of pods (plant-1), number of seeds (plant-1), thousand seeds mass (g), seed yield (g plant-1), harvest index and plant mass (g). The highest seed yield was in the accession MG 101785 (5.33 g plant-1). Seed yield per plant was in the highest positive simple correlations with number of seeds per plant (r = 0.877) and number of pods per plant (r = 0.708). (1) Ellis T. H. N., Poyser S. J., Knox M. R., Vershinin A. V. and Ambrose, M. J. (1998) Molecular Genetics, 260, 9-19. (2) Elvira-Recuenco M. and Taylor J. D. (2001) Euphytica, 118, 3, 305-311. (3) Mihailović V., Mikić A. and Ćupina B. (2004) Acta Agriculturae Serbica, IX, 17 (special issue), 61-65.

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The possibilities of grasspea utilization in foods

J. Rysová1, J. Ouhrabková1, D. Gabrovská1, I. Paulíčková1 and T. Vymyslický 2

1 Food Research Institute, v.v.i., Radiová 7, 102 31, Praha-Hostivař, Czech Republic 2Research Institute for Fodder Crops, Ltd., Zahradní 1, 664 41, Troubsko, Czech Republic [email protected] Grasspea (Lathyrus sativus) is annual plant of the family Fabaceae. It originally comes from India, Pakistan and Nepal. Today is grown in Mediterranean area, in Balkan Peninsula, in the Middle East, in India and in China. It is also important crop in Etiopia (1). Grasspea is able to grow both on poor and on heavy soils. It is resistant to droughts. Grasspea is used both for human consumtion and for fodder. In Europe is often cultivated in organic farming systems. The whole seeds are used after peeling as legume or the seeds are grinded into flour. It is also used as vegetable in some regions. Seeds of grasspea contain much proteins, minerals and TDF. Fat content is mostly low. Seeds consist of carotenoids, vitamins B1, B2 and niacin. Important is calcium and phosphorus content. Seeds of grasspea contain also neurotoxic element beta-N-oxalyl-L-alfa, beta-diaminopropionic acid (β-ODAP). When grasspea is consumed regularly and for a long time it causes dynamics failures (3). Occasional consummation of grasspea combined with other foods is not considered to be dangerous (2). In the frame of the research project NAZV QG 60130 "Minor crops for specific use in food industry" basic nutritional composition of grass pea and beta-N-oxalyl-L-alfa,beta-diaminopropionic acid (β-ODAP) content were determined. The grass pea seeds contained 26,5-28,5% of proteins, 1,3-1,6% of fat, 3,3-3,6% of minerals and 21-29% of TDF. The ODAP content varied from 0,4 to 0,6g/100g of dry matter. The grass pea flour was used into food as a partial substitution of wheat flour. The samples of sourdough breads, gluten free breads, other bakery products and noodles were prepared. Chosen bakery products were evaluated on the basic nutritional compounds content and the sensoric analysis was carried out. The maximal addition of grass pea flour to the wheat flour reached 25% in breads, 20% in cheese straws and 30% in noodles. The amount of grass pea in the gluten free raw materials was 22%. (1) Chowdhury M. A. and Slinkard A. E. (2000) Genetic Resources and Crop Evolution, 47, 163–169. (2) Granati E., Bisignano V., Chiaretti D., Crino P. and Polignano G. B. (2003) Genetic Resources and Crop Evolution, 50, 273-281. (3) Yan Ze Yi et al. (2006) Phytochemistry, 67, 107-121.

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Potential for forage and grain yields in Lathyrus species

Đ. Krstić1, B. Ćupina1, V. Mihailović2, A. Mikić2 and R. Hauptvogel3

1University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia 2Institute of Field and Vegetable Crops, Novi Sad, Serbia 3Research Institute of Plant Production, Piešťany, Slovakia [email protected] The flora of Serbia is rather rich in Lathyrus L. species, but today they are mostly neglected crops. A small-plot trial was carried out at the Rimski Šančevi Experiment Field in 2007 and 2008, including five accessions belonging to yellow vetchling (L. aphaca L.), flat-podded vetchling (L. cicera L.), ochrus vetchling (L. ochrus (L.). DC.), grass pea (L. sativus L.) and Tangier pea (L. tingitanus L.). There were monitored forage yield components, such as plant height (cm), number of stems (plant-1), number of lateral branches (plant-1), yield of green forage (plant-1 and t ha-1), yield of hay (plant-1 and t ha-1) and portion of forage dry matter (%), as well as grain yield components, such as plant height (cm), first fertile node height (cm), number of nodes (plant-1), ordinal number of the first fertile node, number of stems (plant-1), number of pods (plant-1), number of seeds (plant-1), thousand seeds mass (g), seed yield (g plant-1), harvest index and plant mass (g). The highest green forage yields were 54.8 t ha-1 in grass pea, 9.1 t ha-1 in yellow vetchling, 35.8 t ha-1 in flat-podded vetchling, 24.4 t ha-1 in ochrus vetchling and 51.1 t ha-1 in Tangier pea. The highest grain yields were 4587 t ha-1 in grass pea, 511 kg ha-1 in yellow vetchling, 2141 kg ha-1 in flat-podded vetchling, 2961 kg ha-1 in ochrus vetchling and 2312 t ha-1 in Tangier pea.

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Grain legumes technology transfer in Old Europe -

archaeological evidence

M. Ljuština1 and A. Mikić2

1University of Belgrade, Faculty of Philosophy, Belgrade, Serbia 2Institute of Field and Vegetable Crops, Novi Sad, Serbia [email protected] Majority of the grain legume crops cultivated in Europe today originate from the Mediterranean, Near Eastern and Central Asian megagene centres (3). Like other cultivated plants, the present grain legume species were derived from their wild progenitors, such as lentil (Lens culinaris Medik.) from L. culinaris Medik. ssp. orientalis (Boiss.) Ponert. The earliest archaeological evidence of grain legume wild seeds belongs to various sites in Syria, with lentil and bitter vetch (Vicia ervilia (L.) Willd.) dating back as early as Epipalaeolithic (17,000–8,500 BC). Grain legumes had been known to humans before they became grown for diverse purposes. The European hunter-gatherers used various grain legume seeds in their everyday life, as witnessed by the remains of lentil seeds in the cave of Franchthi, Greece, from about 11,000 BC and the seeds of pea (Pisum sativum L.), vetches (Vicia spp.) and vetchlings (Lathyrus spp.) from the site of Santa Maira, Spain, from 12,000–9,000 BP. It is generally considered that the process of domestication led to certain morphological changes in all crops, being essentially similar to the methods of contemporary plant breeding. There are three major criteria used to determine the possible domestication of grain legumes: non-dehiscent pods, larger seed size and smooth seed testa. All three characteristics are often hard to interpret and that remains the main obstacle in bringing forth the evidence that the domestication of grain legumes could predate cereals (2). Among the earliest findings of cultivated grain legumes is the site of Tell El-Kerkh, Syria, from 10th millennium BP, with the seeds of lentil, pea, bitter vetch, chickpea (Cicer arietinum L.), grass pea (Lathyrus sativus L.) and faba bean (Vicia faba L.). The spread of cultivated grain legumes in Europe was associated with the start of the 'agricultural revolution' in the Old World, via Danube and from the Mediterranean coasts (1). Pea, lentil and bitter vetch were found together at numerous sites from early Neolithic, that is, about 6,000 BC, all over Europe. Faba bean was introduced later but became the main pulse in the Mediterranean areas. Grass pea and other vetchlings seem to have been cultivated in the Iberian Peninsula since 7,500 BC. Common vetch (Vicia sativa L.) has been present in the Balkan agriculture at least since Eneolithic (3650-3350 BC) and subsequently, mostly by Roman conquests, was transferred northwards. Lentil was the first grain legume that reached Armenia and Georgia, in 6th or 5th millennium BC. (1) Erskine W. (1998) Use of historical and archaeological information in lentil improvement today. Proceedings of the Harlan Symposium The Origins of Agriculture and Crop Domestication, 10-14 May 1997, Aleppo, Syria, 191-198. (2) Tanno K. and Willcox G. (2006) Veget. Hist. Archaeobot., 15, 197-204. (3) Zeven A. C. and Zhukovsky P. M. (1975) Dictionary of Cultivated Plants and Their Centres of Diversity. Centre for Agricultural Publishing and Documentation, Wageningen, the Netherlands, 219.

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Grain legumes technology transfer in Old Europe -

linguistic evidence

A. Mikić1, M. Ljuština2, V. Đorđević1, B. Ćupina3, V. Mihailović1 and S. Vasiljević1

1Institute of Field and Vegetable Crops, Novi Sad, Serbia 2University of Belgrade, Faculty of Philosophy, Belgrade, Serbia 3University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia [email protected] The European continent may be regarded as rather linguistically rich, at least three hundred living and extinct languages. The most abundant linguistic family of Europe is Indo-European, followed by Uralic, with its Finno-Ugric branch. Along with these two main linguistic families, there are also Altaic, Caucasian, Kartvelian and Afro-Asiatic, while the Basque language is considered a language isolate, with no demonstrable relationship with other languages (2). Majority of annual legumes, that are traditionally cultivated in Europe, originate from the Central Asian, Mediterranean and Near Eastern centres of diversity. In addition to the botanical evidence, archaeology places pea (Pisum sativum L.), bitter vetch (Vicia ervilia (L.) Willd.), chickpea (Cicer arietinum L.) and lentil (Lens culinaris Medik.) among the first domesticated plants in the entire Old World. The fact that Europe with Near East and Northern Africa was home to various ethnic groups throughout the history, as well as that there are common vocabularies of diverse annual legumes to both languages within one and between different families, gives a basis to believe that these crops have been well-known to the ancestors of nearly all modern European nations from time immemorial (3). The Proto-Indo-European language was rich in the roots that gave origin to the words denoting grain legumes in its modern descendants (1), such as *bhabh-, denoting faba bean, *erəgw(h)-, denoting a kernel of leguminous plant, *g'er(a)n-, denoting grain, *ghArs-, denoting a leguminous plant, *kek-, denoting pea, *lent-, denoting lentil, *pis-, meaning to thresh, and *weik-, meaning to avoid. Many of these words were transferred along with crops to the peoples belonging to other linguistic families, such as Uralic. Another testimony of how old crops grain legumes are may be found in the Caucasus region, where several linguistic families have been existing next to each other for millennia and where each of them have its own terms denoting grain legumes. (1) Mikić A., Mikić-Vragolić M., Ćupina B., Mihailović V., Vasić M. and Vasiljević S. (2007) Book of Abstracts of the 6th European Conference on Grain Legumes, Lisbon, Portugal, 12-16 November 2007, 122. (2) Mikić A., Ćupina B., Mihailović V., Vasić M. and Đorđević V. (2008) Proceedings of the 18th EUCARPIA General Congress, Valencia, Spain, 9-12 September 2008, 193-197. (3) Mikić-Vragolić M., Mikić A., Ćupina B., Mihailović V., Vasiljević S., Krstić Đ. and Vasić M. (2007) A Periodical of Scientific Research on Field and Vegetable Crops, 44, II, 91-96.

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Session 3: Status and requirements of legume breeding


Requirements of a practical legume breeding program (faba beans, peas) from the research community

L. Ghaouti and O. Sass

Norddeutsche Pflanzenzucht Hans-Georg Lembke, Hohenlieth, Germany [email protected] From 2003 until 2007, the acreage of legumes in Europe has decreased by about 16% (FAO, 2008) leading most breeding companies to decrease or even to cease their legume breeding activities. Therefore, it is more than ever crucial for the survival of legumes in Europe to maintain a minimum level of breeding activities and maximise the genetic success of such activities. This contribution aims to give an overview of breeding goals in spring and winter faba beans plus spring and winter peas and indicate in which areas recent achievments in research may be applicable and meaningful. The major problem facing faba bean breeding is the low yield stability due to the sensitivity to biotic and abiotic stresses. Biotic stresses comprise mainly diseases which can significantly limit the performance. In spring and winter forms the respective limiting diseases are different (spring pea: Aphanomyces, Ascochyta, winter pea: Ascochyta, spring faba bean: Downy mildew, winter faba bean: Ascochyta). Additionally there are several pests causing problems. Abiotic stresses which are threatening the legumes are drought and for winter sown crops the resistance to harsh winter conditions. For the quality aspect, it is crucial to increase the seed protein content and protein quality. Furthermore, one of the most challenging objectives for faba bean breeders is to overcome the partial allogamy by a stable genetic switch of the mode of reproduction to a full allogamy or a full autogamy. Another critical aspect addressing faba bean and especially pea breeding is to broaden the genetic basis of the European Germplasm that is becoming narrower and in case of peas is already very narrow. The increase of the genetic diversity will allow the introduction of new positive alleles for the relevant agronomic traits. The success of the promotion of legumes in Europe needs the strengthening of the cooperation between the breeders and the research community. However more efforts should be done to address the key topics that are relevant to the breeders and to provide answers that are likely to be adopted directly in the practical breeding. FAO (2008) Food and Agriculture Organization, http://faostat.fao.org/site/567/default.aspx#ancor.

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New interest in grain legumes in Finland

F. L. Stoddard

Department of Applied Biology, PL27 (Latokartanonkaari 5-7), University of Helsinki, 00014, Finland [email protected] After many years of decline in Finland and other Nordic and Baltic countries, interest in grain legumes in Finland is undergoing a resurgence. High fertilizer costs, here as elsewhere, are focusing farmer attention on biological nitrogen fixation. High costs of soybean meal for animal feed are causing feed manufacturers to look for alternatives. Positive drivers include recently released research showing that pigs fed rations containing a substantial proportion of faba beans produced better quality pork meat than those fed on rapeseed meal or soybean meal. As a result, one major manufacturer of animal feed has expressed an interest in contract growing substantial areas of faba bean in Finland for use in pig feed. Another feed manufacturer has quality requirements that closely match lupin grain for ruminant feed. Our results show that the recently released faba bean cultivar 'Jõgeva', from the Jõgeva Plant Breeding Institute in Estonia, matures as successfully in our climate as the older Finnish cultivar 'Kontu'. The larger seeds of cv. Jõgeva may make it more appealing (lower testa content) to the feed compounder and the larger seedlings appeared to be better competitors against weeds in our field trials. Our results also show that several new German cultivars of blue lupin (Lupinus angustifolius) can mature successfully in our climate. The efficient phosphorus nutrition of lupins may add to their attractiveness, as phosphate fertilizer usage is strongly limited within the Baltic basin on account of water pollution of the Baltic Sea. Earliness is a key factor in grain legume adaptation to the Nordic region. Seasons are short and relatively cool. Novel ways of achieving earliness, such as the terminal-inflorescence mutation of faba bean, may have more application here than they have achieved elsewhere. We plan cooperative field trials to further demonstrate the possibilities of grain legumes as a "new" crop, while also researching mechanisms of enhanced adaptation and stress resistance.

50

Session 3: Status and requirements of legume breeding Notes

51


Dry bean (Phaseolus vulgaris L.) breeding in Serbia

M. Vasić1 and M. Zdravković2

1Institute of Field and Vegetable Crops, M. Gorkog 30, 21000 Novi Sad, Serbia 2Institute of Vegetable Crops, Smederevska Palanka, Serbia [email protected] Dry bean have an important place in agricultural production in Serbia. Intercropping with corn cultivars had been the traditional method of bean growing, but nowadays it is grown mostly as a pure stand. The change in growing method required the change in the selection of bean. Cultivars with bushy and erect habit and small, white, round white grains, introduced from the Americas, had been grown at the beginning. These were replaced by new domestic cultivars which differed in grain size, color and shape (1). Presently, there are 15 bean cultivars on the National Variety List, with 14 developed in the domestic research centers (http://www.sorte.minpolj.sr.gov.yu). Unfortunately, because of inadequate advertising and a feeble activity of the extension service, farmers still grow local domestic populations and maintain them by themselves, although they are inferior to the registered cultivars in both seed quality and yield performance. In the Institute for Vegetable Crops in Smederevska Palanka, so far there have been developed the following white-grained cultivars (2): Biser (1980), with erected bushy stem, round grains and grain size of 270 g; Panonski Gradištanac (1984), with branching bushy growth, half-flat grains and grain size of 420 g; Panonski Tetovac (1984), with a slightly longer growing period, kidney-like grain and grain size of 600 g; Galeb (1987), an early cultivar, with irregularly-romboid grain shape and grain size of 400 gr. The assortment contains also Palančki Zlatno Žuti (2005), a medium early cultivar, with loose bushy habitat, old-gold-coloured grain and grain size of 470 g. When the Institute of Field and Vegetable Crops embarked on a program of bean breeding, a decision was made to try and meet the specific market demands. Emphasis was placed on the development of high-yielding varieties, suitable for growing in pure stand and for machine harvesting, but these varieties were also expected to have grain characteristics similar to those of the traditional bean populations (3). Two varieties with colored grains have been released so far. Zlatko, a medium late variety (on the national varietal list since 1994) has large, cylindrical, golden-yellow grains, which retain shape during cooking. Sremac (released in 1999) is an early variety with greenish grains. Determinate white-grained varieties are developed by carefully selecting heterogeneous population over several generations. The first variety to be released (1997), Dvadesetica, is early and high-yielding, with semi-planate, medium large grains (1000/grain mass is about 400 g). The variety Maksa has white, medium large grains, the variety Belko has small, rounded grains. These varieties were entered on the national varietal list in 1998. The variety Balkan has rounded grains with a 1,000-grain mass of over 330 g and was released in 2000. The tetovac-type variety Levač (2004) is characterized by tall, volubile growth. It is not capable of unsupported growth and is intended for growing in a pure stand. (1) Vasić M. (2004) Genetic divergence in a bean collection, Zadužbina Andrejević, Belgrade, Serbia, 94. (2) Zdravković M. (1998) PhD Dissertation, University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia. (3) Vasić M., Gvozdanović-Varga J. and Takač A. (2001) Contemporary Agriculture, 1-2, 237-245.

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Breeding of white clover (Trifolium repens L.) and birdsfoot trefoil (Lotus corniculatus L.) in Serbia

J. Radović, Z. Lugić, D. Sokolović, R. Štrbanović and T. Vasić

Institute for Forage Crops, Kruševac, Serbia [email protected]

White clover and birdsfoot trefoil are widespread perennial legumes, which are very important in providing the protein component in forage grown on the low quality and low pH soils, which is not suitable for alfalfa and red clover. Those forage crops are extremely important for animal husbandry development in mountainous and hilly areas. They are regular components of natural and artificial meadows and pastures in Serbia, especially because they are perennial (over 5 years) and have a good compatibility with other grass components in mixtures. Birdsfoot trefoil and white clover also have a positive effect on the yield and dry matter quality of grass associations (1). In the middle of 80th breeding program on this species were started in Serbia. The main goal of breeding was created genotypes with high production and good herbage quality. The wild and domestic populations of birdsfoot trefoil and white clover, founded in natural pastures in Serbia, were rich sources of genetic diversity for plant breeders and represent excellent initial selection materials for obtaining of high productive and persistent genotypes (2, 3).Until now, as results of breeding process, one cultivar of white clover and four cultivars of birdsfoot trefoil were created in Serbia. There are great possibilities for the further spread of these species in grass -legume mixtures in Serbia. One of the conditions for a faster expansion of birdsfoot trefoil and white clover is a continual improvement of the material available by introduction and selection of new genotypes. The main limiting factors of the currently cultivated species are the high risk of poor stand establishment and the poor stand persistence, especially in mixture with grasses. Slow growth after harvest and tolerance for stress factors in field condition, especially drought tolerance are interesting fields of investigation which could help to improved breeding material. Breeding cultivars which are better adapted for use in grazing situation (with fine steam, prostrate growth…) and cultivars for hay productions which have to be more erect, with establish and faster grow after harvesting, are very important for husbandry development in mountainous -hilly areas in Serbia. Because of that, besides forage yield and forage quality, yield stability, long term persistence in legumes - grasses mixtures, tolerance for grazing and drought will be the main topics for further birdsfoot trefoil and white clover breeding in Serbia. (1) Blumenthal M. J., McGraw R. L. (1999) In: Trefoil: The Science and Technology of Lotus. CSSA, USA, 97-120. (2) Lugić Z., Ignjatović S., Lazarević D., Radović J. (2001) Journal of Mountain Agriculture on the Balkans, 4, 1, 53-62. (3) Radović J., Lugić Z., Dinić B., Ignjatović S., Delić D. (2004): Grassland Science in Europe, 9, 410-412.

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What a seed producer needs from a plant breeder – the example of Novi Sad (NS) forage legume varieties

Đ. Karagić, S. Katić, V. Mihailović, S. Vasiljević, A. Mikić and D. Milić

Institute of Field and Vegetable Crops, Novi Sad, Serbia The NS varieties of forage legumes (lucerne, red clover, pea and vetch) have a satisfying genetic potential for seed yield. However, specific plant morphology leads to extremely great losses in the process of seed production. Breeding should decrease these losses to an acceptable level. The main way of the utilisation of forage legumes in Serbia is the production of voluminous feed (green forage, hay, haylage). By that reason, breeding leads to the development of varieties with rather high yield of forage, high leaf proportion in the total yield and slender and soft stems with high digestibility. Such plant morphology and the chemical composition of cell walls and intercellular space implicates a significant susceptibility to lodging. It is lodging that represents the major problem in the seed production of all forage legumes in the agroecological conditions of Serbia. The seed producers try to delay the beginning of lodging and to decrease its intensity by the regionalisation of production and diverse agronomy practices (mode and time of sowing of vetch and pea, the cutting schedule in lucerne and red clover). However, the effects of these measures are limited and we thus need the varieties of decreased susceptibility to lodging. Breeders should make an acceptable compromise between forage yield and quality at one side and susceptibility to lodging at the other side. The susceptibility to lodging in forage legumes could be decreased by shortening the internodes and increasing the number of nodes. In the NS varieties of protein pea, breeding successfully solved the problem of lodging, plant height was reduced from 130-170 cm to 60-80 cm. However, there appeared a new problem – thin and insufficiently elastic seed coat that easily breaks during maturing and harvest. That is the reason why the viability in the variety Javor is in average about 16 % and 11 % lower in comparison to other pea varieties. The thousand seed mass in the most productive of the NS pea varieties is 261 g (with a range from 233 to 308 g), causing a rather high sowing rate (300-330 kg ha-1). In the conditions of such high sowing rate, the quality of sowing is significantly lower and the cost of sowing is significantly higher in comparison to other pea varieties. Thousand seed mass in protein pea should not exceed 230 g. Increasing the uniformity of flowering in the NS forage legume varieties (especially red clover and vetch) could shorten the period of maturing and significantly decrease seed losses in harvest, thus having a rather positive impact upon the increasement of seed yeilds. Breeding should decrease the susceptibility to pod dehiscence and seed shattering in vetch. Breeding should decrease the susceptibility of protein pea to powdery mildew and aphids in protein pea.

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Breeding pea for grain and fodder in Osijek

R. Gantner1, M. Stjepanović1, S. Popović2 and T. Čupić2

1Faculty of Agriculture in Osijek, Osijek, Croatia 2Agricultural Institute Osijek, Osijek, Croatia [email protected] Field pea for dry grain and fodder is being bred in Osijek since 1987, but this work is intensified since the year 1997. Comprehensive variety trials are being conducted each year since 1997. Varieties of various origins were evaluated. Also they were used for investigation of yield and yield components interaction with aim to define the primary selection objectives (2). Old varieties were used to select the breeding lines too. In the year 1997 there were only 5 registered varieties of field pea for dry grain in Croatia, all of foreign origin, and only 8 fodder varieties; only one of them was of Croatian origin (1). Collection of parental lines is being continuously enriched with introductions of divergent varieties and accessions of ecotypes and wild relatives. Breeding lines and accessions are being utilized in a new variability creation which is followed by selection, mostly pedigree. Most important breeding goals are high yield and quality of grain and fodder and resistance to biotic and abiotic stress. The results of numerous hybridizations and selection are two registered pea varieties: winter fodder pea Osječki zeleni (2002) and spring dry pea Gold (2005), created in collaboration of Agricultural Institute Osijek and Faculty of Agriculture in Osijek. Also, there are three genotypes that are currently being tested by Seed and Seedlings Institute in multi-location state trials. Two of them are of dry pea type and one is fodder type. In two previous years of state trials, both dry pea genotypes yielded better then standard. Average yield of our new genotype in 2006 was 4,6 tonnes per hectare while standard yielded 4,3 tonnes per hectare in average. In 2007 our new genotype average was 5,7 tonnes per hectare while standard yielded 4,7 tonnes per hectare in average. Thus our new genotypes have demonstrated their high yielding potential. (1) Group of Authors (2004) National List of Varieties 2004, Institute for Seed and Seedlings, Osijek, Croatia (2) Čupić T., Popović S., Tucak M., Grljušić S., Stjepanović M. and Andrić L. (2005) XL Croatian Symposium on Agriculture, Opatija, University of J. J. Strossmayer in Osijek, Faculty of Agriculture, 170-180.

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59


World chickpea collection variety testing in mountainous

regions of the Republic of Armenia

N. Sarukhanyan1 and A. Vanyan2

1Green Lane Agricultural Assistance NGO, Yerevan, Armenia 2Agricultural Reform Support Project Implementation Unit SI, Ministry of Agriculture of RA, Yerevan, Armenia [email protected] In Armenia, legume crops mostly cultivate local populations carrying the name of the given place or area and which are more adaptive to soil and climatic conditions. It needs to be noted anyhow that the populations cultivated are not homogeneous and contain diverse forms. This kind of diversification among populations creates difficulties in cultivation of legumes, generation of high yields, as well as in the introduction of new technologies. The generation and introduction of new, more productive varieties is henceforth more apt to time demands. Upon careful review of the issues highlighted above, the “Green Lane” NGO has been successfully running a series of research activities aimed at the application of new cultivation technologies for bean, chickpea, lentil and peanut, testing of new varieties’ and their introduction, as well as weed, pest and disease prevention and control measures. The main cultivars produced in Sissian region of Syuniq Marz are cereal crops. In the last years a drastic fall in their productivity was recorded, caused by the lack of crop rotation, as well as the use of old agricultural machinery, poor-quality seedling, reduced soil productivity and other factors. Taking into account that chickpea is ranked as a drought- and frost-resistant crop among other legumes, we considered it wise to organize their cultivation in Sissian region. The variety “Leninakanskaya 313” and other 6 varieties selected from 120 species out of the world chickpea collection served as control samples for the use in trial plots. The mentioned varieties were offered by the Scientific Center of Agriculture and Plant Protection. The morphological structure of seed varieties shows that they are different in heights of the plants, ramification (branchiness), dimensions of leaves, and the form of their grains and seeds. The transition periods of plants’ phenostages were studied during the vegetation period. The varieties were specified according to their productivity levels. As compared to non-studied varieties, those studied gave high yields in droughty conditions. The varieties tested, in relation to control samples, provided an incremental yield of 7,8-23,5 %. The varieties “Lilit” 19.7, “Alina”19.1, “Sissian”18.0 and “Hacavan”18.6 c/ha showed enhanced productivity in droughty conditions of the RA mountainous regions . The tested varieties are bushy, well-stalked, straight and medium-sized. “Lilit”, “Alina”, “Sissian” and “Hacavan” are proven to be the most productive varieties recommended to be introduced into production . The varieties “Alina” and “Hacavan” stand out with improved taste properties for grain. The varieties “Alina”, “Sissian” and “Hacavan” appeared to be highly resistant to diseases, particularly to askokhitoz The varieties tested are recommended for cultivation in droughty conditions of the RA mountainous regions. Thus, we suggest farms not only to focus on their cultivation, but to scale up further to seed production.

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Theoretical foundations and practical results of adaptive

soybean selection in Ukraine

V. F. Petrychenko, S. V. Ivanyuk and S. I. Kolisnyk

Feed Research Institute of the Ukrainian Academy of Agrarian Science, Vinnitsa, Ukraine [email protected]

Evolution of cultivated crops and transformation of ecosystems involved into the sphere of production are the unity of two system components, interaction of which is a motive force for the improvement of functionality of integral agrophytocenosis of agricultural crops including soybean (1, 2). Taking into consideration the origin of this crop (the Far East, monsoon climate), soybean cultivation had substantial problems in Ukraine that has temperate continental climate. Due to absence of adapted soybean varieties in Ukraine areas of soybean cultivation could not exceed 100,0 thousand ha, the main areas were located in the Steppe of Ukraine. By the 90th of the XX century in Ukraine 8 soybean varieties were zoned and only 1 variety could be cultivated in conditions of the Forest-Steppe. During the last decades more than 6 Ukrainian scientific-research institutions have selected nearly 80 high productive adapted domestic soybean varieties, including more than 15 varieties selected under support of Feed Research Institute of UAAS with seed yield of 3,5-4,5 t/ha. Among them are Oksana, Anatolievka, Podilska 416, Monada, Omega vinnitskaya, Appolon, Medeya and others. More than 65% of all domestic varieties belong to early or mid ripening groups (3). Research of the world soybean collection in conditions of soybean cultivation in Ukraine and selection of parent forms for hybridization has positive results. Great amount of soybean varieties from Canada, the USA, China and Russia were used as initial material for selection of high productive mid ripening varieties of Ukrainian selection. Varieties from the Western Europe, Germany, Hungary, and France were used in saturant crosses when selecting early ripening soybean varieties. Availability of these soybean varieties of different genetic nature demands scientifically substantiated approach to the distribution of variety sowings in various regions of Ukraine. This investigation is based on the idea of soybean belt of Ukraine, main point of which is in construction of adaptive biosystems of each region of soybean cultivation taking into account increased potential productivity and ecological resistance of plants (4). (1) Litup P. P., Kirichenko V. V., Petrenkova V. P. and Kolomatskaya V. P. (2007) Adaptivnaya selektsia: Teoriya i tehnologia na sovremennom etape, 263. (2) Petrychenko V. F., Babych А. О., Ivanyuk S. V., Kolisnyk S. І. (2006) Agrarna nauka/Visnyk agrarnoi nauku, 2, 19-23. (3) Derzhavny Reestr sortiv roslyn prydatniy dlya poshyrennya v Ukraini na 2008 rik (2008) 59-62. (4) Recomendatsii schodo rozrobky tehnologichnogo protsesu vyrobnytsva soi na bogarnyh zemlyah/Pid red. V. F. Petrychenka (2007) 15.

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Participatory Plant Breeding (PPB) approach for evolving

superior varieties of soybean (Glycine max L. Merril) under maize based system in Nepal

R. Darai1, B. N. Adhikari2, R. K. Neupane1, B. S. Bastola2 and B. R. Baral3

1Regional Agriculture Research Station/Nepal Agricultural Research Council, Nepalgunj, Khajura, Banke, Nepal 2National Grain Legumes Research Program, Rampur, Chitwan, Nepal 3National Oil Seed Research Program, Nawalpur, Sarlahi, Nepal [email protected]

Soybean is a wonderful legume crop of Nepal. Being high nutritious legume crop helps to food & nutritional security and sustainable soil management in the system. Soybean yellow mosaic virus, pod blight, bacterial pustule, frogeye leaf spot, rust, white fly and hairy caterpillars are the major menacing of the production. Unavailability of suitable variety for various cropping patterns such as Maize-soybean-toria /other winter crops, Rice + Soybean (Bund Planting) has been realized in the recent years. Released varieties have more or less smaller seed size, susceptible to different foliar diseases and longer maturity period (135-145 days), which farmers usually don't prefer. In the present context due to the increasing trends of poultry farming in the country, the demand of soybean is sharply increasing. In connection with above background soybean genotypes collected from diversified sources were evaluated in observation screening nursery (OBN) and multi-location trial for yield potential, insect-disease reaction in the research station of Rampur and Nawalpur during 2004 to 2006. Multi-location trial was carried out in RCB design with four replications and OBN in rod row. No of rows of the genotypes were two in OBN, four in multi-location trial. Some lines AGS124 (2719 kg/ha), SB0065 (2589 kg/ha), G1873 (2482 kg/ha) and LS7716-16 (2371 kg/ha) were selected from observation screening nursery based on the over years performances and will be promoted in multi location trial in next season. Similarly, the genotypes Puja (1699kg/ha), TGX-311-23D (1691 kg/ha) and PK-327 (1614 kg/ha) were found outstanding performances than the standard check cobb (1367 kg/ha) and significant differences among the genotypes over the year and across location in multi location trial. Therefore, these lines will be evaluated in farmer's participatory trial. Participatory research is the spirit of the time for dynamic and effectively adaptation to the farmers root level. Maintaining biodiversity, selection of lines according to farmer and user's choice, farmer-to-farmer seed diffusion leads to fast adoption and rapid impact at farm level. In the brightness, farmers preferred Puja for it's earlier maturity (125 days), resistant to yellow mosaic virus and bacterial pustule, higher yield (yield potential 2.5 t/ha), medium bold seed size (13.83g/100 seed) and adjust well in maize based system either in mono or intercrops. It is also well adapted from terai/inner terai to mid hills environment. National Seed Board has released it for general cultivation.

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High-precision breeding: reliable and cost-efficient

molecular tools are looking for users

P. Winter GenXPro GmbH, www.genxpro.de, Altenhöferallee 3, D-60438 Franfurt, Germany [email protected] Recent advances in all areas of molecular breeding as e.g. comparative genomics, or SuperSAGE whole-genome transcription profiling provide unprecedented insights into molecular components underlying traits in non-model crops. Novel tools based on these insights such as custom gene-expression and SNP-detection micro-arrays, and qRT-PCR assays to detect SNPs and differential expression of genes in response to developmental signals and in response to the environment have been developed also in legumes and are available to everybody. We will give an overview of progress in understanding biotic stress responses of major European legumes to fungal pathogens and demonstrate the power of SuperSAGE for the identification of potential breeding targets for dehydration stress tolerance in chickpea. Downstream of SuperSAGE, fast, reliable and cost-efficient qRT-PCR assays indicative for stress-response pathways offer themselves for application in knowledge-based breeding. Especially, a molecular definition of trait components available in hitherto under-exploited wild and cultivated germplasm that could be introgressed with as yet unprecedented speed and accuracy could pave the way to new varieties better suited to changing market needs. Now, intense communication between molecular biologists, plant physiologists and breeders is required to drive molecular approaches into application-oriented directions. This presentation is meant to elicit such communication.

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Genetic analysis, QTL mapping and gene expression analysis of key visual quality traits affecting the market

value of field pea

L. Ubayasena1, T. Warkentin1, K. Bett1, B. Tar'an1 and D. Bing2 1Crop Development Centre, University of Saskatchewan, 51 Campus Drive, Saskatoon, Saskatchewan, Canada S7N 5A8 2Lacombe Research Station, Agriculture and Agri-Food Canada, Lacombe, AB, T4L 1W1, Canada [email protected] Visual quality is one of the major factors determining the market value of field pea (Pisum sativum L.). Breeding for improved visual quality of pea seeds is currently a challenging task, mainly because of the complexity and the lack of sound genetic knowledge of the traits. The objectives of this research were to characterize the genetic basis and conduct QTL mapping of four key quality traits (cotyledon bleaching in green pea, greenness in yellow pea, seed shape, and seed dimpling in both green and yellow types). Gene expression profiling to understand the molecular basis of post-harvest cotyledon bleaching in green pea was addressed in this project. Two F5:6

recombinant inbred line (RIL) populations (93 lines from Orb X CDC Striker cross, and 120 lines from Alfetta X CDC Bronco cross) were developed and evaluated for the quality traits in two locations in Saskatchewan, Canada in 2006 and 2007. The four quality traits evaluated all displayed a continuous range of expression among the relevant RILs, with moderate to high heritability. Two genetic linkage maps utilizing 201 markers (30 SSR (from Agrogene) and 171 AFLP markers) and 155 markers (21 SSR and 134 AFLP markers) were constructed for Orb X CDC Striker and Alfetta X CDC Bronco populations, respectively. Three major QTLs on linkage group (LG) IV and one QTL on LG V for green cotyledon bleaching resistance were identified. Three QTLs on LG I and IV using Orb X CDC Striker population and one major QTL on LG VII with two additional QTLs on LG VII and I using the Alfetta X CDC Bronco population associated with seed shape were recognized. Three QTLs on LG I and IV associated with seed dimpling were recognized using the Orb X CDC Striker population, while no significant QTLs were detected on the linkage map of Alfetta X CDC Bronco population for this trait. Two major QTLs associated with greenness in yellow pea seeds on LG I and II were recognized. The bleaching resistant cultivar CDC Striker had a slower rate of chlorophyll degradation in cotyledons, and a higher carotenoid to chlorophyll ratio in seed coats, than the bleaching susceptible cultivar Orb when seed samples were exposed to high intensity light. An oligo-nucleotide microarray (Ps6kOLI1) was utilized to study the molecular expression of bleaching resistance in pea seed coats. The gene expression profiles of the CDC Striker and Orb seed coats were significantly different during the seed developmental stages at 14 and 21 days after flowering (DAF). A significant up-regulation of genes involved in the production and accumulation of secondary metabolites which are responsible for antioxidant properties in plants tissues such as kaempferol 3-0 glucoside, quercetin 3-0 glucosides, pentunidin 3-0 glucoside, and malvidin 3-0 glucosides, in the seed coats of CDC Striker at 14 and 21 DAF were observed.

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Linkage between genes for leaf size, leaf shape, stipule

size, pod size and globe plant type in lentil (Lens culinaris Medik.)

Y. Kumar1, S. K. Mishra1, M. C. Tyagi1, A. Sarker2 and B. Sharma1

1Division of Genewtics, Indian Agricultural Research Institute, Delhi 110012, India 2International Center for Agricultural Research in the Dry Areas, P.O. Box 5466, Syria [email protected] Linked genes with monogenic inheritance have been discovered for leaflet size (broad vs. narrow leaflets. Blf), leaflet shape (oval vs. acute, Ol), stipule size (large vs. small, Lst), pod size (large vs. small, Lpd), and globe mutant (normal vs. globular plant canopy, Glo). Broad leaflet was dominant over narrow leaflet, oval leaflet over acute, large stipule over small, large pod over small pod, and normal plant type over globe mutant. Linkage was established from the F2 segregation analysis of 2247 plants in six crosses for leaflet size, 2239 plants in six crosses for leaflet shape, 2140 plants in six crosses for stipule size, 2512 plants in seven crosses for pod size, and 1127 plants in two crosses for globe plant type under field conditions. taking two characters at a time in all possible combinations when a pair of traits did not show independent assortment. Based on crossing over values the genes were arranged in the short linkage sequence Lst–Ol–Blf–Lpd–Glo. This short sequence of linked genes has been called linkage group III (LG III) of lentil. The genetic map of these five genes can be represented as follows:

18.9

12.5

12.4

Lst 9.6 Ol 8.9 Blf 6.2 Lpd 9.6 Glo

16.2

33.9

25.4

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RAPD markers in estimating diversity of soybean parental

genotypes and predicting traits of progeny

V. Perić, S. Mladenović-Drinić and M. Srebrić Maize Research Institute „Zemun Polje“, 11185 Zemun-Belgrade, Serbia [email protected] Breeding self-pollinated crops is based on developing and identifying superior genotypes, which result from transgressive segregation of parental alleles. If the parents are genetically distant and differ for more genes that affect the trait, the probability of obtaining such genotypes increases. Genetic markers are useful tool for estimating genetic diversity between parents and predicting the performance of traits in progeny (1, 2). Objection of this study was to estimate genetic distance (GD) between genotype ZPS 015 and genotypes L 91-3103, Ravnica, Shine, L 90-14 and ZPS 107, and compare GD to seed yield and number of superior lines of their progeny. Genotype ZPS 015 has been used as a parent in crosses with other five genotypes. RAPD analysis of these genotypes with 18 random primers gave in total 75 fragments which of 20 % were polymorphic. Coefficients of similarity were calculated by Jaccard, Rogers and Tanimoto, Sorensen-Dice and Sokal and Michener. All coefficients showed small genetic distance between parental lines. Low level of polymorphism and genetic divergence between parental pairs is in accordance with previous analyses about polymorphism of soybean, as a species with very narrow genetic background (3). In order to compare genetic distance between parents and seed yield’s data of F4 progeny by each cross, Spearman's Rank Correlation Coefficient was calculated. This coefficient showed negative and non-significant correlation. (1) Kisha T. J., Sneller C. H. and Diers B. W. (1997) Crop Sci., 37, 1317-1325. (2) Manjarrez-Sandoval P., Carter T. E., Jr., Webb D. M. and Burton J. W. (1997) Crop Sci., 37, 698-703. (3) Nikolić A. (2005) MSc Thesis, University of Belgrade, Faculty of Biology, Belgrade, Serbia, 53.

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In vitro regeneration of African yam bean (Sphenostylis stenocarpa (Hochst et A. Rich) Harms.) by direct

organogenesis

A. I. Adesoye and A. Emese Department of Botany and Microbiology, University of Ibadan, Nigeria [email protected] African yam bean (Sphenostylis stenocarpa) is a hardy protein-rich underutilized tropical legume that has been proven to have high nutritional value (1). The lack of acceptance and adoption over the years may, in addition to its hard seed coat which makes it difficult to cook, be attributed to the presence of anti-nutritional secondary metabolites that are known to be present in seed and vegetative parts of this plant (2). Also the high incidence of seed borne fungal pathogen have been reported to significantly reduce seed germination, seedling emergence, as well as the nutritional qualities of the seeds (3). Therefore in vitro micropopagation for the production of clean and disease free plants is essential. Some workers (4) reported conditions for successful in vitro germination of seedlings and sterilization of explants for tissue culture studies. However, there is no report yet on in vitro morphogenesis in this crop. Studies on in vitro organogenesis of African yam bean (Stenostylis stenocarpa) were carried out with the aim of developing a rapid regeneration system for this crop. Shoot tips, cotyledonary nodes, embryos and immature leaf explants were cultured on Murashige and Skoog (MS) containing varying concentrations and combinations of 6-Benzyl aminopurine (BAP); 6-Furfuryl aminopurine (KN) and α-Naphthalene acetic acid (NAA). The highest number of embryo explants with multiple shoots (20) and number of shoots (20) was produced on media containing 0.5 mg/l BAP and 0.05 mg/l NAA. Maximum shoot length was obtained on media with 1.0mg/l KN and 0.1mg/l NAA. Embryo explants showed significantly higher multiple shoot response with BAP+NAA combinations than KN+NAA (p<0.05) treatments. When cotyledonary node and shoot tip explants were cultured on media with BAP and KN singly each at 1.0, 1.5 and 2.0 mg/l both explants produced maximum number of shoots and shoot length at 2.0 mg/l BAP while the least responses were obtained at 1.0 mg/l KN. There was no organ formation from immature leaves as they all produced calli. Multiple shoots from embryo produced roots directly on shoot induction culture while shoot tip -derived multiple shoots rooted when transferred to 0.025 mg/l NAA. Shoots from cotyledonary nodes did not. Successfully rooted plantlets were obtained from this study and this is the first report of in vitro plant regeneration in AYB. Direct organ differentiation would facilitate micropropagation and improvement of this species through genetic transformation. (1) Azeke M. A. (2004) PhD Thesis. Bonn, Germany, 177. (2) Asuzu A. and Undie A. (1986) Quality of Plant Foods and Human Nutrition, 36, 3-9. (3) Nwachukwu G. O. and Umechuruba C. I. (1997) Global Journal of Pure and Applied Sciences, 3, 2, 143-147. (4) Aliyu R. and Adesoye A. (2007) Journal of Biological Science Research, 2, 1, 36-39.

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Quantitative Real-Time-PCR: fast and efficient technology

for knowledge-based breeding

D. Steinhauer1, F. Khan2, N. Fatnassi3, C. Molina4, R. Horres1, C. Abdelly3, M.-J. Delgado5, G. Kahl4 and P. Winter1

1 GenXPro GmbH, www.genxpro.de, Altenhöferallee 3, D-60438 Franfurt, Germany 2 Jamia Hamdard University, New Dehli, 110062 India 3 Laboratoire d’Adaptation des Plants aux Stress abiotiques, Centre de Biotechnologie de Borj Cedria, BP 901 2050 Hammam-Lif, Tunisia 4 Biocentre, University of Frankfurt, Germany 5 Estacion Experimental del Zaidin c/Prof. Alboreda, 18008 Granada, Spain [email protected] Dehydration stresses such as drought and salinity constrain yield of chickpea throughout the world. Using SuperSAGE whole-genome transcription profiling, we analysed more than 600.000 root- and nodule transcript SuperTags from salt-and drought tolerant chickpea varieties and compared them to non-stressed control plant to identify potential molecular targets for knowledge-based stress-tolerance breeding. Amongst expression profiles differentiating between tolerant and susceptible varieties those involved in signal perception and transduction and also transcripts coding for down-stream effectors such as reactive-oxygen-(ROS)-related proteins provided interesting candidates deserving further evaluation for their value in breeding. However, not only transcripts coding for proteins of potentially known function but also transcripts coding for proteins of unknown function as well as their different splicing variants were differentially expressed in tolerant susceptible varieties. For example, SuperTag STCa-4092 encoding the putative seed storage protein Narbonin belonged to the most up-regulated transcripts under both salt and drought stress. Using the 26bp SuperTag as specific primer for 3’- and 5’-RACE, we sequenced 4 different splicing variants of the respective mRNA from chickpea roots. The protein translated in-silico from the largest (5-exon) variant was highly homologous to the Narbonin protein sequence and to the PFAM Glyco_hydro_18 domain (E-value 5.10e-08) and eventually able to form the Triosephosphate Isomerase and Concanavalin B-like structure of the crystallized Narbonin protein, whereas the 3-exon-variant solely encoded the glycoside hydrolase catalytic core. We used commercially available, splicing-variant-specific quantitative Real-Time (qRT)-PCR assays to investigate the temporal expression of the different splicing variants in stressed and non-stressed roots and nodules of salt- and drought tolerant and susceptible chickpea varieties. All splicing variants were higher expressed in at least 1 tolerant variety as compared to susceptible varieties already under non-stress conditions. Further, whereas in tolerant varieties all splicing variants were generally up-regulated under stress, susceptible varieties reacted by their down-regulation. The fact that putative Narbonin transcripts were already much stronger expressed in tolerant genotypes than in susceptible ones even before the onset of stress suggests a necessity of “priming” for stress-tolerance. Therefore, it may be possible to pre-screen germplasm and breeding lines for stress-tolerance using qRT-PCR even without stressing it.

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Session 4: Fundaments of legume breeding Notes

79


From lab to field: the case study of molecular tools for marker assisted breeding in pea virus and fungal

resistances

P. Smýkal Agritec Plant Research Ltd., Plant Biotechnology Department, Šumperk, Czech Republic [email protected] Crops start to benefit from available genomic knowledge of model plants. In addition, large body of information is generated in academic research. However for application, numerous constrains still limit the use of such data, one of them being broader genotype range testing and verification. Such work is find not to be of great scientific interest, not being worth of high impact publishing. Leaving the space for applied, agronomy oriented research. The incorporation of marker-assisted selection (MAS) into breeding strategies would result in reduction in the number of offspring to be propagated, selected and tested. While traditional plant breeding relies upon crosses and subsequent selection of genotypes to meet desirable traits such as yield, quality and disease resistances. MAS currently leans on statistical testing of associations between DNA markers and chosen trait and only in few cases when specific allele of gene of interest was identified, direct assisted selection of desirable combinations of genes is possible. In case of pea (Pisum sativum L.), the testing of resistance to viruses such as Pea Seed-borne Mosaic Virus (PSbMV) and fungus, powdery mildew (Erysiphe pisi) pathogens, is included in breeding process. The resistance to PSbMV is conferred by a single recessive sbm-1 or er-1 locus in case of mildew, localized on LG VI and IV respectively. For sbm-1 locus, sequences derived from the gene for the eukaryotic translation initiation factor eIF4E and amino acid exchanges were already identified (Gao et al. 2004), while for er-1 only indirect, linked SSR markers exist (Ek et al. 2005). We have analyzed eIF4E genomic sequences from further 47 pea varieties and breeding lines, reported as donors of resistance. This analyses led to identification of novel alleles. Subsequently, PCR-based, both single nucleotide polymorphism (SNP) and co-dominant amplicon length polymorphism markers were developed and proved to be fully reliable on broad spectra of pea varieties and breeding lines. For er-1, use is limited to crosses where parents are polymorphic for given SSR loci together with 98% maximal accuracy of scoring. All these data will be discussed in light of breeding process applications, aiming at selection efficiency improvement by circumvention of the need for laborious and time consuming symptomatic testing. Acknowledgements: This work was financially supported by Ministry of Education of the Czech Republic research project MSM 2678424601 and INGO LA08011. Gao Z et al. (2004) Theor. Appl. Genet., 109, 488-94. Ek et al. (2005) Hereditas, 142, 86-91.

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Session 4: Fundaments of legume breeding Notes

81


Differences in seed proteins among high-protein soybean

genotypes

K. Taški-Ajduković1, V. Ðorđević2, M. Vidić1, J. Miladinović2 and M. Vujaković1 1National Laboratory for Seed Testing, Maksima Gorkog 30, 21000, Novi Sad, Serbia 2Institute for Field and Vegetable Crops, Maksima Gorkog 30, 21000, Novi Sad, Serbia [email protected] Soybean (Glycine max L. (Merr.)) became the most important vegetable source of proteins, hence a requirement to generate soybean genotypes with a higher protein content and quality. Forty soybean genotypes developed at the Institute for Field and Vegetable Crops in Novi Sad, Serbia were used in the study. Nine of them were newly developed genotypes, and one standard cultivar from each maturity groups: 0, I, II and III. The protein content of the most tested new soybean genotypes (except KO5319) in all maturity groups was significantly higher as compared to the normal seed protein cultivar used as check cultivar for proper maturity group. Soybean storage proteins have two major fraction, β-conglycinin (7S) and glycinin (11S), accounting for approximately 70% of the total proteins (1). Because of their abundance, β-conglycinin and glycinin are mainly responsible for soybean protein quality, thus the relative accumulation of these subunits is estimated by scanning densitometry (2). Our study confirmed observation that high-protein cultivars accumulated higher amount of glycinin and β-conglycinin. Genotypes KO5427, KO5428, and KO5429 that accumulated lower amount of all subunits of glycinin and β-conglycinin comparing to the check cultivars were the only exceptions. Attention should be placed on genotype KO5314, which accumulated significantly higher amount of the both subunits of glycinin, and genotypes KO5425, KO539, and KO536 that accumulated significantly higher amount of β-conglycinin subunits compared with the check cultivar. All these findings suggest that some of the tested genotypes could be beneficial in breeding programs aimed at altering composition of seed storage proteins. (1) Hill J. E. and Breidenbach R. W. (1974) Plant physiology, 53, 742-746. (2) Taški-Ajduković K., Ðorđević V., Vidić M., Vujaković M., Milošević M., Miladinović J. (2008) Genetika, 40, 9 -16.

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Characteristics of special importance in red clover

(Trifolium pratense L.) breeding

S. Vasiljević1, Z. Lugić2, G. Šurlan-Momirović3 and M. Ivanović1

1Institute of Field and Vegetable Crops, Novi Sad, Serbia 2Institute for Forage Crops, Kruševac, Serbia 3Faculty of Agriculture, University of Belgrade, Belgrade, Serbia [email protected] In addition to the classic selection targeted on the development of high-yielding red clover cultivars resistant to economically important diseases and possessing satisfactory quality, new breeding objectives have been defined which are aimed at the development of highly adapted cultivars specialized for growing in particular regions. Regarding the latter set of objectives, it is particularly important to evaluate initial breeding material for the morphological and biological characteristics (date of flowering, persistence, growth type) that indirectly or directly affect the forage yield performance and crop persistence. Flowering is a complex characteristic which is affected by both genetic and environmental factors. Beginning of flowering has always been considered as an important parameter in red clover because it served to classify cultivars into early, mid-season and late ones. Furthermore, correlations have been found between the flowering of red clover in the year of planting and poor persistence (the so-called two-cut «medium» type), and between a lack of flowering in the year of planting and good persistence (the so-called one-cut «mammoth» type (1). Besides genetic factors, flower initiation is largely affected by environmental factors, especially light duration. Poor persistence of red clover (Trifolium pratense L) is a major problem that prevents further expansion of this forage legume important for the production of quality feed (2). This problem has always been paid due attention. Causes of inferior persistence of red clover have never been adequately explained, although suspicion has been placed on fungal diseases, insect attacks and unfavorable weather conditions. The inferior persistence of early types has been associated with reduced capacity of the plant to survive the internal breakdown of crowns. Particularly important for survival of plants that suffered the internal breakdown of crowns is the development of an adventitious root system. It is of interest to note that the numerous investigations of this phenomenon failed to isolate pathogens that could be suspected as potential causes of s crown necrosis. Histological investigations have indicated that a possible cause could be an anomaly of cell nuclei within the crown tissues. A somewhat higher red clover persistence has been found in the prostratum type. Recent investigations have indicated that in vitro selection is closely associated with increased tolerance of red clover to stress conditions caused by low temperature, i.e., with improved persistence (3). The term “growth habit” is variously interpreted by different authors. Coulman and Oakes (1987) were among first to propose a classification of red clover growth habit on a scale from 0 to 4, which fully complies with a widely accepted classification of red clovers to two basic types, “mammoth”, a one-cut type which only forms the rosette in the first year, and “medium”, a two-cut type brings forth flowers in the first year. According to the former classification, 0 stands for red clover genotypes which form the rosette but do not flower in the first year, while 4 stands for red clover genotypes which flower but do not form the rosette in the first year of cultivation. Although it had been assumed that the genotypes classified in group 0 were characterized by superior winter hardiness, it was found that improved persistence, i.e., overwintering capacity, was not invariably correlated with a high proportion of plants that did not flower in the first year of cultivation. According to UPOV descriptor from 2001, red clover growth types include five

86


different types, visually rated on a scale of 1 to 9: (1 - erect, 3 - semi-erect, 5 - intermediate, 7 - semi-prostrate, 9 - prostrate). The descriptor further maintained that the prostratum type was more persistent than the erectum type, and that it could be used for crossing to commercial cultivars of the erectum type, to increase their persistence.

(1) Coulman B. E. and Oakes B. (1987) Canadian Journal of Plant Science, 67, 1, 276-277. (2) Cressman R. M. (1967) Crop Science, 7, 357-361. (3) Nowak J., Matheson S. L., McLean N. L. and Havar. P. (1992) Euphytica, 59, 2-3, 189-196. Notes

87


Combining ability of pea genotypes

R. Gantner1, G. Drezner2, M. Stjepanović1, S. Popović2 and T. Čupić2

1Faculty of Agriculture in Osijek, Osijek, Croatia 2Agricultural Institute Osijek, Osijek, Croatia [email protected] Knowing combining ability of parental lines is important in estimating a potential of a parent in a new variety creation. The general opinion is that parents with good combining ability will give a good new genotype. F1 generations with high values of desired traits often have high probability of giving more valuable progeny in later generations. Aim of this research is to estimate general combining ability of 6 parental lines of field pea based on 6×6 diallel cross without reciprocal crosses. General combining ability of each parent was calculated as an average value of progeny of all parental combinations containing respective parent. There were tested 15 F1 combinations of progeny of diallel cross on traits of plant height and grain yield per plant in 2008 year. Parental lines were spring field pea varieties as follows: Danish yellow dry pea Anno, medium tall (about 52 cm) with afila leaf type, Croatian yellow dry pea Gold with low plant habit (about 48 cm tall) and afila leaf type, American yellow dry pea Shawnee with high plant habit (about 84 cm tall) and wild leaf type, American green dry pea Joel with high plant habit (about 83 cm tall) and wild leaf type, new yellow dry pea genotype PF-G1, creation of Faculty of Agriculture in Osijek and Agricultural Institute Osijek, with medium high plant habit (about 65 cm tall) and wild leaf type and Serbian field pea NS-Junior, arvense type for fodder and grain (yellow grain) production with high plant habit (about 118 cm tall) and wild leaf type. F1 progenies were seeded in optimal agrotechnical term (12th March 2008) with stand of about 100 seeds per square meter in microplots, in Osijek, on eutric cambisol. Upon harvest, there were determined plant height and grain yield per plant. The general combining ability of each parent for the plant height trait was as follows: Gold 66,06 cm, Anno 66,41cm, PF-G1 78,66cm, Shawnee 87,60cm, Joel 88,91cm and NS-Junior 96,34 cm. For the grain yield per plant the general combining ability was as follows: Gold 6,64 g, Shawnee 7,00 g, Anno 7,26 g, Joel 7.51 g, PF-G1 7,71 g and NS-Junior 7,95 g. NS-Junior has performed the best combining ability for grain yield per plant and high plant height. However, association of high yield with high plant height is considered as unfavorable in grain production due to plant lodging potential and excessive biomass that impedes harvesting. If in segregating generations the NS-Junior's progeny fail to segregate separately regarding these traits, the best genotypes will probably be obtained of other good combinators.

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89


Farmer-participatory selection and scaling up lentil

varieties in Nepal

R. Darai1, A. Sarker2, N. K. Yadav3, D. B. Gharti3, B. N. Adhikari3 and D. N. Pokhrel1

1Regional Agriculture Research Station/NARC, Nepalgunj, Khajura, Banke, Nepal 2ICARDA, Aleppo, Syria 3National Grain Legumes Research Program, Rampur, Chitwan, Nepal [email protected] Lentil (Lens culinaris Medikus subsp. culinaris) is the main pulse crop of Nepal which shares about 60 % of a dozen of pulses grown in Nepal. Being an exportable commodity, its share in the world export market is about 2% (FAO, 2004). In addition to human food, lentil straw is used as animal feed and it plays a vital role in improving soil fertility and thus makes farming systems sustainable. In the past, classical plant breeding has benefited farmers of favorable environments and those who can afford inputs and have access to irrigation. Participatory varietals Selection (PVS) is considered a technical strategy to conserve local genetic resources by adding values to them and promote it in the potential areas. Indeed PVS helps to empower the farmers and new cultivars will be scaled-up and extend to newer areas. Maintaining biodiversity, selection of lines according to farmer and user's choice, farmer-to-farmer seed diffusion lead to fast adoption and rapid impact at farm level (Sarker et al., 2003). A number of lentil lines were evaluated under PVS program in intensive lentil-growing areas during winter seasons of 2004 and 2005 in Devnagar, Pithuwa and Kumroj villages of Chitwan and Purena village of Banke district in Terai region of Nepal. Selection based on plant and seed traits, maturity, and yield after harvest done by farmers, researchers and extension worker resulted into identification of promising lines: ILL7982 (1083 kg/ha), ILL4402 (957 kg/ha) and ILL 7164 (929 kg/ha) over the years and across locations. Pooled analysis of genotype, ILL7982 was superior to the check shital and other tested genotypes which produced 11.54% higher yield than the check. Community based seed production and distribution systems have started in the project sites. Farmer to farmer seed distribution was found encouraging for self-reliant of seeds.

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Conventional breeding of Kunitz-free soybean genotypes

M. Srebrić and V. Perić

Maize Research Institute, Slobodana Bajića 1, Beograd – Zemun, Serbia [email protected] The soybean grain is important source of plant protein, especially for animal feeding, but it also contains undesirable components. Mature grain of conventional soybean varieties requires heat processing to break down trypsin inhibitor’s activity before using as food or animal feed. At the same time, protein denaturation and other qualitative changes occur in soybean grain, especially if the temperature of heating is not controlled. Kunitz-trypsin inhibitor (KTI), is a predominant antinutritional factor, which makes 30-50% of trypsin inhibitor’s activity. KTI is present at a concentration of 1·4 g/kg of total seed contents (2). Trypsin inhibitors inhibit activity of trypsin. The ingestion of raw soybean meal by monogastric animals causes protease’s hyper production and pancreatic hypertrophy, at the same time growth rate decreased. Mature grain of conventional soybean varieties requires heat processing to break down trypsin inhibitor’s activity before using as food or animal feed. At the same time, protein denaturation and other qualitative changes occur in soybean grain, especially if the temperature of heating is not controlled. Four types of KTI have been identified in soybean grain. The three types (Tia, Tib, and Tic) are inherited as codominant alleles in a multiple allelic system at a single locus . The absence of KTI is inherited as a recessive allele and has been designated ti. (3). Variety Kunitz (1), lacking KTI, has been used, as a parent donor of desirable character. Kunitz-free lines were developed from a few cross combinations between Kunitz soybean wariety and adapted high yeailding genotypes. Progenys were slected and presence of KTI was tested into a few segregating generations. Two methods were used for identification Kunitz-free individual plants: gel electrophoresis and Hamerstand and Black & Glover method. Genotypic×locational interaction, and fertilization were found to be significant for the whole trypsin inhibitor activity in Kunitz-free and standard quality soybean grain(4). Soybean grain, without KTI can be processed at a lower temperature and for a shorter period of time. In that case, we save energy, and preserve valuable nutritional composition of soybean grain, which is of interest in industrial processing. This trait makes them also suitable for direct feeding in adult non ruminant animals without previous thermal processing. (1) Bernard R. L., Hymowitz T. and Cremens C. R. (1991) Crop Science, 31, 232-233. (2) Clarke E. J. and Wiseman J. (2000) The Journal of Agricultural Science, 134, 125-136 (3) Orf H. J. and Hymowitz T. (1979) Crop Science 9, 107-109.

(4) Vollman J., Grausgruber H., wagentristl H., Wohleser H.and Michele P. (2003) Journal of Science of food and Agriculture 83, 1581-1586.

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Theoretical preconditions for the development of effective methods for selection of self-incompatible genotypes of

Medicago sativa L.

V. D. Bugayov, A. M. Maksimov and V. S. Zadorozhnyi Feed Research Institute of the Ukrainian Academy of Agrarian Science, Vinnitsa, Ukraine [email protected] It is well known that the highest level of heterosis effect can be achieved in controlled cross-pollination of correspondingly chosen parent forms. One of the mechanisms of genetic control of lucerne cross-pollination is the use of self-incompatibility (1). When selecting lucerne on basis of self-incompatible genotypes, one of the most difficult moments is the process of selection from the initial populations which demands much checking work over several years. It is especially important to estimate self-incompatibility taking into account different types of geitonogamy pollination. As a result of researchers (2002-2007) the nature of lucerne self-incompatibility manifestation as a result of selection of the best genotypes for further selection of synthetic varieties has been studied. It is determined that variability of self-incompatibility in investigated populations has more than 20% variation. Distribution in population has exponential character with modal class in the interval of 90-100%, which makes up from a third to a half of the whole population. Plants with stable manifestation of this trait make up 3.9% (3). Correlation dependence of self-incompatibility level and pollen fertility level within -0.434 up to -0.481 is proved. Substantial impact of pollination type on the nature of correlation dependence of beans’ number per hundred of flowers depending on the level of self-incompatibility is revealed. In self-pollination within a flower connections were close (R2>0.49). In geitonogamy pollination connections had medium power (0.09< R2<0.49), and in cross-pollination connections were weak (0.09< R2 or R2≈0.09), negative in direction. It is determined that self-pollination mainly affects seed reductions per bean in comparison to cross-pollination. It is shown that with the rise self-incompatibility level abortion of seed germs of lucerne ovary falls down. Besides abortions of absolutely self-incompatible genotypes were less by 1.5-3 than in plants with self-incompatibility level of 60.0-79.9% (2). According to generalized results of researches effective method of selection of self-incompatible plants has been developed. It enables to identify genotypes with the stable manifestation of self-incompatibility level. The main point of this method is in the use of self-pollination – fusion of gametes produced by one flower. Thus, selected self-incompatible plants are to be checked additionally by means of geitonogamy pollination which enables to reveal stably self-incompatible plants during the year of trial (4). Genotypes with the increased level of self-incompatibility (70-100%) were singled out using this method in order to get synthetic varieties among which 79 are self-incompatible.

(1) Campbell T. A. and He Y. (1997) Canad. J. Plant Sc., 77, 1, 69-73. (2) Bugayov V. D., Maksimov А. M. (2005) Kormy i kormovyrobnytsvo, 55, 9-15. (3) Maksimov А. M. (2007) PhD Thesis. Vinnytsia, Ukraine, 113. (4) Pat. №17756 Ukraine, MPK А01N 1/04. Sposib vydilennya samonesumisnyh lyutserny posivnoj; Zayav. U 2006 03646; Opub.16.10.2006, Byul., 10, 4.

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Genetic and phenotypic correlations among morphological

and agronomic traits of lucerne genotypes

D. Milić, S. Katić, A. Mikić and S. Vasiljević

Institute of Field and Vegetable Crops, Novi Sad, Serbia [email protected] During 2001-2003, 30 lucerne genotypes of different geographic origin were studied at Rimski Šančevi over seven cuts. The genotypes were broadcast-planted into hills 0.8 x 0.5 m in size in 2001. Ten agronomically and morphologically important traits were studied: yields of green forage and dry matter, plant height, number of shoots (stems) per plant, internode number and length, length and width of the middle leaf blade, proportion of leaves in the yield, and stem diameter. Genetic and phenotypic correlations among the morphological and agronomic traits of the genotypes were calculated. Genetic correlations had higher values than the phenotypic ones. The coefficients of correlation between the yields of green forage and dry matter were highly significant (rg=0.98), as the level of functional dependency was nearly achieved. Significant positive correlations were found between green forage yield and shoot number (rg = 0.83) and dry matter yield and shoot number (rg =0.76). These traits can be used in tandem selection. Also, highly significant positive coefficients of genetic correlations were found between plant height and internode length (rg = 0.87), indicating that genotype had a significant influence on the expression of these traits. Highly significant negative correlations were found between leaf contribution to yield and green forage yield (rg =-0.81), dry matter yield (rg =-0.93), and plant height (rg = -0.91), which is in agreement with Julier et al., (2000). Breeding for a higher proportion of leaves in yield will result in a higher quality of green forage and dry matter, but the yields will be lower. Phenotypic correlations among morphological and agronomic traits showed significant positive interdependence between the yields of green forage and dry matter (rf = 0.97), shoot number and dry matter yield (rf= 0.76), and plant height and dry matter yield (rf = 0.66). On the other hand, negative correlations were found between leaf proportion in yield and forage yield, plant height, and internode length. In Katić (2001), the values of phenotypic correlations were higher than those of genetic ones, which may have been due to the greater influence of environmental factors resulting from the fact that the plants were grown in a dense stand as opposed to being broadcast-planted individually, as was the case in our study. Julier et al. (2000) reported obtaining genetic and phenotypic correlations similar to ours when broadcast sowing was used. Studying the effects of genetic and environmental factors on yield and yield components is of great importance in lucerne breeding, as is the knowledge of genetic and phenotypic correlations among lucerne traits and their practical application (especially in tandem selection). (1) Julier B., Huyghe C. and Ecalle C. (2000) Crop Science, 40, 365-369. (2) Katić S. (2001): PhD dissertation, University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia.

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Grain yield components in afila (af) lines of field pea (Pisum

sativum L.)

V. Mihailović, A. Mikić, S. Katić, Đ. Karagić and I. Pataki

Institute of Field and Vegetable Crops, Novi Sad, Serbia [email protected] The discovery of the recessive gene af controlling the transformation of all leaflets into tendrils (afila) in field pea is often considered the most important event in the field pea breeding (1). Such cultivars are characterized by a significantly improved standing ability, non-decreased photosynthetic activity and often higher grain yields in comparison to the cultivars with normal leaf type (2). A small plot trial has been carried out at the Experiment Field of the Institute of Field and Vegetable Crops at Rimski Šančevi between 2006 and 2008, including 12 afila lines of field pea. The study included plant height (cm), number of nodes per plant, number of fertile nodes per plant, number of pods per plant, number of grains per plant, thousand grains mass (g) and grain yield per plant. The average grain yield per plant varied from 2.15 g in the line AF09 to 7.78 g in the line AF02. Grain yield per plant was in the highest positive simple correlation with number of pods per plant (r = 0.951). Thousand grains mass was the only grain yield component that was in negative simple correlations with all other grain yield components, most notably with number of nodes per plant (r = -0.668). (1) Duparque M. (1996) Grain Legumes, 12, 18. (2) Mihailović V. and Mikić A. (2004) Genetika, 36, 1, 31-38.

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Challenges in breeding vetches (Vicia spp.) for diverse

purposes

R. Matić1, S. Nagel1, G. Kirby1, I. Young2 and K. Smith3 1South Australian Research and Development Institute (SARDI), Adelaide, Australia 2Westminster School, Adelaide, Australia 3University of Adelaide, Adelaide, Australia [email protected] Among the economically most important species of the genus Vicia L. are faba bean (V. faba L.), common (V. sativa L.), hairy (V. villosa Roth), Hungarian (V. panonnica Crantz), bitter (V. ervilia (L.) Willd.) and Narbonne vetches (V. narbonensis L.). All these species may be cultivated as field crops in diverse forms for animal feeding, such as green forage, hay, silage, haylage, grain and straw (2). At the same time, all these species are one of the most valuable crops for green manure and thus have a significant role in organic farming and sustainable agriculture (1). The modern breeding programmes on common, Hungarian and hairy vetches are aimed at the improvement of forage, grain or biomass yields, the improvement of resistance to major diseases such as rust and ascochyta, the facilitation of the adaptation to low-rainfall areas and the screening for possession of soft seeds, non-shattering pods and high grain and hay quality (3). Every yield is a trait of low heritability, conditioned by a large number of genes. The standard breeding methods are used for intra-species gene recombination and include pedigree and back crossing. These methods are often combined with recombined-delivered family (RDF) methods, that speed up elimination of off-type plants in F2 and F3 generations and that are used to identify superior progeny for further back crossing, testing and selection. The advantage of the RDF methods is that low-yielding individuals and families are rapidly eliminated from the testing programme, while, on the other hand, these methods reduces diversity from gene recombination: the method works well for plant height and maturity in the generations from F2 to F4, but does not provide satisfactory expression of disease resistance or yield stability. Since Hungarian vetch is a species without any known toxin in its grain, it may be used for reciprocal crossings with common vetch. However, these crossings do not produce viable seeds. If common vetch is used as a female parent, pods develop and abort 1-3 weeks after the crossingl if it is used as a male parent, pods develop with viable seeds that reach about 1/4 of their full size but finally abort; further attempts should involve embryo rescue methods. Among the most important achievements of the contemporary common vetch breeding are potential for a hay crude protein content up to 260 g kg-1, a dry grain crude protein content of about 300 g kg-1, less than 0.60 % of grain toxins and an increase of total nitrogen up to 200 kg ha-1. (1) Ćupina B., Erić P., Mihailović V. and Mikić A. (2004) A Periodical of Scientific Research on Field and Vegetable Crops, 40, 419-430. (2) Matić R., Nagel S., Robertson S., Young I., Mihailović V., Mikić A. and Kirby G. (2005) Biotechnology in Animal Husbandry, 21, 5-6, 2, 203-207. (3) Matić R., Stuart N., Kirby G., Young I. and Smith K. (2007) A Periodical of Scientific Research on Field and Vegetable Crops, 40, II, 55-63.

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In search of a low toxin feed vetch

M. E. Tate, D. Chowdhury and H. Firincioglu

School of Agriculture, Food and Wine, University of Adelaide, Adelaide, Australia [email protected] Aim: To ultimately breed lines of Vicia sativa (common vetch) with less than 0.4% g-Glu-b-CN-Ala + b-CN-Ala for poultry-feed. Conclusions: 1. Best selection to date is a dark seeded Love 3 line (0.45 +/- 0.07%).

2. Use of white testa for distinguishing low toxin lines. 3. Selection for Low toxin (LO-VETTM) Vicia sativa cultivars is facilitated by Single seed analyses of the first two seeds in a pod, using SAS JMP Self Organising Map clustering and Discriminant Analysis software.

102

Love.3 (S-H) W

Love.2 (S-H) W

Morava (Field) D

Rasina (Field) D

J.White (Field) W

IR28 (S-H) D

Morava (S-H) D

Languedoc (S-H) D

J.White (S-H) W

Languedoc (Field) D

Bl. fleur (Field) D

Love.3 (Field) D

Love.1(Field) D

Au

str

alia

n V

.sa

tiv

a C

ult

iva

rs

.0 .2 .4 .6 .8.9 1.1 1.3 1.5 1.7 1.9 2.1

Mean(Quadratic Corrd % Toxin)

Chart


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103


Bulk segregation analysis and improvement of seed quality

in soybean

V. Đorđević, J. Miladinović, S. Balešević-Tubić, V. Đukić and A. Mikić Institute of Field and Vegetable Crops, Novi Sad, Serbia [email protected] One of the goals in soybean breeding program is increasing protein content. On LG I was located major QTL for protein content, originate from Glycine soja (1). This protein QTL was detected in some Glycine max genotypes, mostly non-adapted germplasm which are inadequate for commercial breeding programs. Goal of this work is to find suitable source for marker assisted selection of soybean to increase seed protein content. A total of 155 F2 plants of soybean, were developed from a cross of Senka x Novosadjanka used in this study. Base on a previous work, can suppose variety Senka is donor parent of protein QTL from LG I (2). Bulk segregation analysis was conducted base on protein content and tested for differences in allele frequency between the population bulks whit microsatellite marker Satt239. This marker is closely linked whit protein QTL from Glycine soja (3). Each bulk contains 12 high protein and 12 low protein F2 soybean lines. There is observed highly-significant differences in allele frequencies between two bulks and strong derogation from expected ration of 1:2:1 for unlinked markers. Alleles segregating in a ration of 1:1 between two bulks and most frequent allele in high protein bulk originate from donor variety Senka. Bulk segregation analysis implies thin linkage between Satt239 and QTL for protein content. Average difference of protein content between two bulks was 3.8%. There is also observed well documented negative correlation between protein and oil content. Oil content was 2.3% higher in low protein bulk. QTL from LG I also have influence on other seed traits, such 1000 seed mass (3). Seed size was also significantly different between two bulks. These results show potential of molecular markers in soybean breeding programs. Future research will show reaction of this QTL in different genetic backgrounds and environments and usefulness in soybean breeding. (1) Chung J., Babka H., Graef G., Staswick P., Lee D., Cregan P., Shoemaker R. and Specht J. (2003) Crop Science 43, 3, 1053. (2) Đorđević V. (2006) MSc Thesis. University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia. (3) Nichols D., Glover K., Carlson S., Specht J. and Diers B. (2006) Crop Science, 46, 2, 834.

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Evaluation of yield potential, genetic variances and correlation for nine cultivars of alfalfa under the New

Valley environment

M. M. Abdel-Galil and N. M. Hamed

Forage Crops Res. Dept., Field Crops Res. Inst. ARC, Giza, Egypt [email protected] In the period of 2004-2006, a field trail was conducted at the New Valley Res. Station to evaluate the yield potential and genetic variances among alfalfa cultivars; a new Synthetic (Wady Syn.), four promising populations (Serw1, Serw2, Nitrogen fixation and Salt tolerant), three commercial varieties (Siwa, Ismailia1 and Ismailia94) and a local cultivar (Wady). Twenty cuts were obtained during 2005 and 2006. The combined analysis of variance over two years indicated that Wady Syn. Population ranked first for fresh and dry yields ( 72.3 t fed-1 and 18.9 t fed-1) and other studied traits were significantly different from other tested cultivars. The commercial variety Ismailia94 ranked second (66.55 and 17.2 t fed-1 ). Regarding to plant height, tillers and leaf to stem ratio (LSR), Wady Syn. recorded the highest values (48.2 cm, 416.7/m2 and 47.6 %) significant from Ismailia94 which recorded 45.6 cm, 362.3 m2 and 43.3 %, respectively. Significant positive correlation among either fresh forage yield or dry forage yield and other traits were found. The values of genotypic coefficient of variation for fresh and dry forage yields revealed relative variations among the tested cultivars which were less influenced by environment. The environmental variation ranged from 4.4% to 33.3% and the genetic advance ranged from 3.9% to 14.5%.

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Developing a synthetic population through selection in Egyptian clover genotypes (Trifolium alexandrinum L.)

M. M. Abdel-Galil, A. A. Helmy and N. M. Hamed

Forage Crops Res. Dept., Field Crops Res. Inst., ARC, Giza, Egypt [email protected] Selection and random cross pollination for fresh and dry forage yields and protein content within seven local Egyptian clover genotypes (Trifolium alexandrinum L.) was applied for two generations to develop a synthetic population. Syn1 and Syn2 were tested against the base populations and two imported Italian varieties (Nilodi and Sacromente). Enormous improvement was achieved where Syn2 had higher values in all studied traits than the best parent (Gemmeza-1), Syn1 and the imported varieties. The realized gains from selection ranged from 13.2- 34.4% for fresh forage yield, 11.4 to 37.7% for dry forage yield and from 2.8 to 8.9% for protein percentage. Heritability in broad sense were high for seasonal fresh and dry yields (88.7 and 88.2%) and 65.0% for protein percentage. The environmental variations were 11.3, 11.7 and 35.4% for fresh and dry yields and protein percentage. The expected genetic advance for fresh, dry yields and protein percentage were 8.1, 2.3% and 0.8% respectively.

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New insights in legume breeding: plant genetics of beneficial plant-microbe systems, evolution and

applications in sustainable agriculture

A. Borisov1, E. Ovchinnikova1, T. Nemankin1, O. Grishina1, O. Shtark1, G. Akhtemova1, A. Krasheninnikova1, A. Moloshonok2, V. Zhukov1, A. Kazakov1, T. Naumkina2, A.

Vasilchikov2, V. Chebotar1, V. Gianinazzi-Pearson3 and I. Tikhonovich1 1All-Russia Research Institute for Agricultural Microbiology, Podbelsky chaussee 3, Pushkin 8, St. Petersburg, 196608, Russia 2Institute of Grain Legumes and Groats Crops, Orel, p/b Streletskoe, 303112, Russia 3UMR INRA 1088/CNRS 5184/Université de Bourgogne PME, BP 86510 - 21065 Dijon Cedex - France [email protected] Existence of common legume plant genes implicated in interactions with both arbuscular mycorrhizal fungi and beneficial rhizosphere bacteria (including nodule bacteria) opens a new hypothetical view to evolution of beneficial legume-microbe systems. Acceptance of such a view point creates a theoretical basis for exploitation of such a plant-microbe system in sustainable agriculture. Great genetic variability by effectiveness of such a system was demonstrated for pea (Pisum sativum L.), and therefore, possibility and necessity of doing breeding to improve symbiotic potential of legume crops were clearly shown. This, in its turn, poses a question of development of new types of complex inoculants to select highly symbiotically effective plants during breeding process. The field trials (performed during years 2000-07) demonstrated high beneficial effect of such a kind of complex inoculation on plant biomass production and protein content in the seeds. Exploitation of such systems in agriculture will allow to decrease application of mineral fertilizers and chemical means for plant protection and will improve quality of agricultural produce.

This work was supported by the grants of RFBR (07-04-01171, 07-04-01558, 06-04-89000, 07-04-13566), NWO 047.018.001, Burgundy administration 07.9201 AA040 S3623, Grant to support leading Russian science school 5399.2008.4, Governmental contracts for research of Russian Ministry of Science and Education (02.512.11.2182, 02.512.11.2254).

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Effect of bean inoculation with Rhizobium leguminosarum bv. phaseoli on pod and grain number and grain mass

J. Marinković1, M. Vasić1 and M. Jarak2

1Institute of Field and Vegetable Crops, Novi Sad, Serbia 2Faculty of Agriculture, Novi Sad, Serbia [email protected]

The use of microbial fertilizer free provides cost-free plant nutrition with atmospheric nitrogen, increases yields and grain protein quality due to the increased presence of essential amino acids, and enriches the soil with nitrogen to be utilized by the subsequent crops. Bean is a crop grown for its highly nutritious grain, which is one of the richest sources of vegetable protein in the human diet and one of the most widely used food items overall. For this reason, it is eseential to work on improving bean grain quality, most importantly grain protein content. The objectives of this study were to examine four strains of Rhizobium leguminosarum bv. phaseoli (5b, 3b/2, B2, P4) on two bean cultivars on the basis of pod and grain number and grain mass. Field trials were established on a chernozem soil at the experiment field of the Institute of Field and Vegetable Crops. Two bean cultivars were tested (Belko and Sremac), developed at the Institute of Field and Vegetable Crops in Novi Sad. Immediately before planting, bean seeds were inoculated with four Rhizobium leguminosarum bv. phaseoli strains. The stady included 5 variants (4 variants – seeds inoculated with Rhizobium leguminosarum bv. phaseoli strains and non-inoculated bean seeds). Plant samples were taken at the end of bean growing season. The obtained results were statistically processed by the analysis of variance and tested by the LSD test. The results showed that the inoculation significantly affected the symbiotic association parameters studied. Number of pods demonstrates the success of pollination of a genotype and thus determines yield. Number of grains per plant is a highly inherited trait and depends on environment to the same extent as on the genotype itself. The cultivar Belko had significantly higher values of numbers of pods and grains per plant, as well as grain mass per plant, in all four inoculated variants in comparison to the control variant. The greatest number of pods was in the variant inoculated with the strain Rhizobium leguminosarum bv. phaseoli 5b (12.20 pods), while the greatest number of grains (40.50 grains), as well as the largest grain mass (10.73 g) per plant as in the inoculation with the strain Rhizobium leguminosarum bv. phaseoli P4. The cultivar Sremac had significantly greater number of pods in the variants inoculated with the strains Rhizobium leguminosarum bv. phaseoli B2 and P4, while the greatest number of pods was in the variant inoculated with the strain P4 (12.07 pods). The inoculation with the strains 5b, B2 and P4 significantly increased number of grains per plant, while grain mass was significantly increased by the inoculation with the strains 5b and B2. The greatest number of grains (36.08 grains) and grain mass (14.31 g) per plant were recorded in the variant inoculated with the strain B2. These results emphasize the necessity of further study of applicability of Rhizobium leguminosarum bv. phaseoli strains for seed inoculation because these microorganisms not only reduce the use of mineral nitrogen fertilizers but also increase soil quality, soil fertility, by provoking an increased production of plant hormones and enzymes.

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Genetic resistance to Phoma medicaginis in pea

K. McPhee1 and X. Wang2

1USDA-ARS, P.O. Box 646434, Pullman, WA 99164-6434, USA 2Crop and Soil Sciences Department, 201 Johnson Hall, Washington State University, Pullman, WA 99164, USA [email protected] The Ascochyta blight complex affecting pea is comprised of three pathogens, Ascochtya pisi, Mycosphaerella pinodes and Phoma medicaginis var. pinodella (1). P. medicaginis var. pinodella causes a blackening on stems near the soil surface and has been referred to as black stem rot or Phoma foot rot. Genetic resistance to P. medicaginis is available in Pisum germplasm; however, resistance has been incorporated into few modern cultivars. A detached leaf assay was used in our laboratory to identify variation for resistance to P. medicaginis var. pinodella. Thirty-five registered cultivars, and breeding lines were evaluated for disease development. Significant differences in lesion expansion were observed and ranged from 2.6 to 173.1 mm2 9 days after inoculation. These results indicated that genetic resistance is present in available germplasm. Parents of several recombinant inbred line mapping populations were included in the germplasm screen and Shawnee and Bohatyr, the parents of PRIL-12 developed susceptible and resistant reactions, respectively. Bohatyr had a mean lesion size of 4.3 mm2 while Shawnee had a mean lesion size of 170.6 mm2. One hundred eighty-seven RILs from PRIL12 were screened in replicate using the detached leaf assay and lesion size ranged from 0.06 to 27.48 mm2 10 days after inoculation. Bohatyr showed a resistant reaction with a mean lesion size of 0.41 mm2 while Shawnee maintained a susceptible reaction with a mean lesion size of 14.11 mm2. Area under the disease progress curve (AUDPC) was also calculated and used for QTL analysis. A genetic map of PRIL12 was developed and comprises 8 linkage groups (LG) and aligns with the consensus Pisum map (2). The eighth LG is comprised of markers on the lower portion of LGIII containing Le and is separated from the remainder of LGII due to lack of sufficient marker density. QTL analysis was conducted based on lesion size 10 d after inoculation and AUDPC values for 178 RILS using QTL Cartographer v. 2.5 (3). One QTL on LGVI was detected based on data for lesion size 10 d post inoculation and AUDPC with a LOD scores of 9.0 and 7.8, respectively. Two smaller QTL each with a LOD score of 2.2 were detected on LGIII. Two additional minor QTL were detected on LGVII with LOD scores of 1.7 and 2.1. Though they appear to be minor QTL, they may contribute toward resistance. Results from this QTL analysis require cross validation in additional mapping populations; however, the presence of a single strong QTL indicates that resistance should be highly heritable and genetic gain from selection is possible. (1) Kraft J. M. and Pfleger F. L. (2001) Compendium of Pea Diseases and Pests, American Phytopathological Society, St. Paul, USA, 67. (2) Loridon K., McPhee K., Morin J., Dubreuil P., Pilet-Nayel M. L., Aubert G., Rameau C., Baranger A., Coyne C., Lejeune I. and Burstin J. (2005) Theoretical and Applied Genetics, 111, 1022-1031. (3) Wang S., Basten C. J. and Zeng Z.-B. (2001-2003) Windows QTL Cartographer 2.5., Department of Statistics, North Carolina State University, Raleigh, USA (http://statgen.ncsu.edu/qtlcart/WQTLCart.htm)

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The expression profiles of selected genes in leaves of different beans species (Phaseolus spp.) under drought

stress

V. Meglič1, T. Kavar1, M. Maras1, M. Kidrič2 and J. Šuštar-Vozlič1

1Agricultural Institute of Slovenia, Hacquetova 17, SI-1000 Ljubljana, Slovenia 2Jožef Stefan Institute, Jamova 39, SI-1000 Ljubljana, Slovenia [email protected] Phaseolus species were known to differentiate for drought tolerance. In order to compare the response to drought stress in leaves of P. coccineus, P. lunatus and P. acutifolius with previously studied P. vulgaris, relative gene expression analysis using qPCR have been carried out for one putative DREB gene and 15 dehydration-responsive transcripts, which had been confirmed as up- or down-regulated in leaves of P. vulgaris previously. Ten transcripts and the reference gene (actin) were successfully amplified in all three species by primers previously developed for P. vulgaris; one transcript (DD5) was amplified only in P. coccineus and P. acutifolius; and five transcripts (CG10, CA1, CG20, 25CA145 and DD8) did not amplify in any species. Relevant homologs of transcripts CA1 and 25CA145 were amplified using novel primers, designed according to P. coccineus sequences. For these 13 transcripts, expression profiles of Phaseolus spp. analysed, were congruent with the P. vulgaris profiles. Seven transcripts were thus up-regulated under drought stress, i.e. for two transcription factors from AP2/EREBP family (DREB and CA1), group III LEA protein (DD5), abscisic stress ripening-like protein (CG18), aldehyde dehydrogenase (CA7), hydrolase, hydrolyzing O-glycosyl compounds (CC3) and ankyrin-kinase (DD19). Six transcripts were down-regulated, i.e. for germin-like protein (Sch-frg), conserved hypothetical protein (DD7), two chlorophyll a/b-binding proteins (25CA145 and NV) and small and large subunit of ribulose 1,5-bisphosphate carboxylase/oxygenase (CG03 and CG05). Similar transcripts were reported in other plant species under conditions of drought stress, thus it is not surprising that the same response was found in all Phaseolus spp. analysed.

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Systemic and local regulation of legume nodulation

P. M. Gresshoff, A. Miyahara, S. Nontachaiyapoom, A. Indrasumunar, Y.-H. Lin, D. Reid, M. Kinkema, B. Biswas, Q. Jiang, M. Miyagi, D. Li, M.-H. Lin, B. Carroll,

P. K. Chan, T. Hirani, A. Kereszt and B. Ferguson Australian Research Council Centre of Excellence for Integrative Legume Research The University of Queensland, St. Lucia, Brisbane QLD 4072, CILR, ANU, Canberra, ACT 2600, Australia [email protected]

Nodule formation in legumes (3) is minimally controlled at two levels: a) the locally-acting perception of the mitogenic signal from the Rhizobium bacterium and b) the systemically-acting Autoregulation of Nodulation (AON), which involves closely related CLAVATA1-like LRR receptor kinases (GmNARK/LjHAR1/MtSUNN) expressed in phloem of most legume tissues; yet its biological activity in nodulation is almost exclusively within the leaf (1,5). To dissect AON, the soybean NARK promoter was analysed to reveal tissue expression domains and a putative phloem specifying promoter domain. Purified kinase domain of GmNARK was able to autophosphorylate and transphosphorylate itself as well as the newly discovered GmKAPP (kinase associated protein phosphatase (2)). A bioassay for the AON shoot-derived inhibitor was developed to indicate that the inhibiting principle was extractable, Bradyrhizobium-induced, NARK-dependent, RNAase A and Proteinase K resistant, and of small size. Down-stream molecular events from GmNARK revealed coordinate expression of genes leading to the synthesis of jasmonic acid. Mutations in LjHAR1 also lead to inhibited main root growth, which was reversed by ethylene insensitivity (ETR1 controlled) in shoots, suggesting a dual role in nodulation and root growth. Increased nodulation, nitrogen gain and ability to nodulate effectively at sub-optimal Bradyrhizobium titres were achieved after overexpression of the GmNFR1α gene. Cloning of GmNFR1 and GmNFR5, complementation of mutants (4), and nodulation efficiency analysis suggest that the AON circuit acts by perception of the NF signalling cascade and subsequently targeting its efficiency. (1) Kinkema M., Scott P. and Gresshoff P. M. (2006) Funct. Plant Biol., 33, 770-785. (2) Miyahara A., Hirani T. A., Oakes M., Kereszt A., Kobe B., Djordjevic M. A. and Gresshoff P. M. (2008) J. Biol. Chem., 283, 25381-25391. (3) Beveridge C. A., Mathesius U., Rose R. and Gresshoff, P.M. (2007) Curr. Opin. Plant Biology, 10, 44-51. (4) Kereszt A., Dong-Xue L., Indrasumunar A., Nyugen C., Nontachaiyapoom S. and Gresshoff P. M (2007) Nature Protocols, 2, 948-952. (5) Nontachaiyapoom S., Kinkema M., Scott P. T., Schenk P. M., Men A. E. and Gresshoff P. M. (2007) Mol. Plant Microbe Interact., 20, 769-790.

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Nitrogen fixation efficiency of several rhizobial strains in symbiosis with Vigna angularis (L.) Willd., Vigna radiata (L.) Wilczek and Vigna mungo (L.) Hepper under field

conditions

D. Delić, O. Stajković, Đ. Kuzmanović, N. Rasulić, S. Maksimović and B. Miličić Institute of Soil Science, 11000 Belgrade, Serbia [email protected] Vigna angularis (L.) Willd., Vigna radiata (L.) Wilczek and Vigna mungo (L.) Hepper are a significant annual leguminous species in tropical and sub-tropical regions. They are used as cheap protein-rich nourishment, primarily as human food, but also as forage and green manure. It is predominantly the grain that is consumed, but green mass and immature pods as well. They have high content of crude proteins (23-28%), carbohydrates (>50%) and mineral matter, as well as low content of fats (<1,5%) in the grain. In symbiosis with specific nodule bacteria Bradyrhizobium spp. they fix molecular nitrogen from the atmosphere. Seed inoculation with Bradyrhizobium spp. bacteria prior to sowing allows a reduction in N mineral fertilization and decreases susceptibility to environmental stress. The characteristics of these leguminous plants indicate a need to introduce them into agricultural production in Serbia. The aim of our research was to investigate the possibility of cultivation of these Vigna species as newly introduced crops in Serbia, by applying seed inoculation with highly effective rhizobial strains in the form of N microbial fertilizer. The effects of seed inoculation with four active rhizobial strains both individually and in combination with mineral N were estimated on V. angularis, V. radiata and V. mungo. Field experiment organized in a randomized complete block split–plot with three replications was performed in two different types of soils. The effect of seed inoculation on grain yield, grain crude protein (CP) content and grain CP yield was done. Inoculation with some rhizobial strains resulted in a significant increase in grain yield for all Vigna species tested. The average increase due to inoculation was 15-40% over the uninoculated control. Inoculation resulted in higher nitrogen uptake. The strain 542 was highly effective with all investigated species, while other strains differed in their effects depending on plant species. The highest grain yield of V. mungo and V. radiata was achieved by inoculation with some strains in combination with 40kg/ha. The results indicate that effective strains may be applied as N microbiological fertilizer to soils in Serbia.

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Legumes seed inoculation advance technology for

biological nitrogen fixation improvement

A. Korakhashvili Georgia National Academy of Sciences, Tbilisi, Georgia [email protected] One of the most important roles in the Food Security Program of Georgia is attributed to the provision of the population with proteins of plant origin. The source of such proteins in our country, from times immemorial was grain legumes such as chickpea, faba beans, lentils, and in the latest time – haricot and soybeans. These crops had been one of the principal foods for Georgians and other peoples residing on our country. The heightening of efficiency of these crops is based on the original method of inoculation of legumes crop seed, elaborated and patented by us, one of the rings of agro-technology. This method enables us to get maximum harvest of grain, as well as straw, at the least expenditures. At the application of this method the seed prior to its sowing does not require chemical treatment, germination rate increases, high expenditures for mineral and organic fertilizers, for Rhizobium bacteria stems, inoculums, energy carriers etc. decrease. Our technology was used in inoculation of legumes plant seed by the method of formation of pill. By our patented technologies (1) seed pills are made as follows (example): 100 kg conditional seed of lentil is covered by 12,9 liter water, it is kept in a closed vessel for one hour up to complete consumption of that water; then seed is taken from the vessel (seed surface must be dry), is put into rotating drum, is added 0,941 l of 30% polyvinyl alcohol water solution and is mixed for 15 minutes, for equal wetting, it is added by boric acid – 0,0134 kg, zinc sulfate– 0,0087 kg, ammonium molibdenate – 0,024 kg, cement –4,3068 kg and Rhizotorphine –0,3529 kg and is mixed for final fixation of component mixture to the seed, for what 10-15 minutes are sufficient. The obtained pill is dried through warm air flood (30-35oC) up to drying the seed surface. Then the seed is rinsed in chloroform, 3% solution of polycarbonate and is dried anew in a flush of warm air (30-35oC) for 10-15 minutes up to complete evaporation of the solvent and the appearance of a film on the seed pills. Especially well were the matters in farm economies distributed in the arid zone of East Georgia, in last 2008 year in Mtskheta, Gurgaani and Dedoplitskaro districts. In these regions, on the land plots of farmers' households, at about 3,8 t/ha chickpea, 2,7 t/ha lentils, and 4,2 t/ha faba bean was obtained on the small trials, while in West Georgia, in the humid zone (Lanchkhuti district) soybeans reached 2,5 ton per ha. The fact is to be emphasized that advantages of elaborated technologies were so apparent and reliable that they found great popularity among farmers. Heightening of efficiency of the results of research works is a demand of contemporary and it must be supported by creation of necessary terms for strengthening of private farm economies of EU countries. This will become a foundation for successful implementation of the Food Security and Safety Programs of our continent. (1) Korakhashvili A. (1996) Georgia State Patent # 1180, Tbilisi, Georgia, 5.

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Progress in pea breeding for disease resistance at Córdoba,

Spain

D. Rubiales1, M. Fernández-Aparicio1, A. Moral1, A. Pérez-de-Luque1, E. Barilli1, J. C. Sillero2, A. M. Torres2, M. Curto3, M. A. Castillejo1, E. Prats1 and S. Fondevilla3

1Institute for Sustainable Agriculture, CSIC, Apdo. 4084, 14080 Córdoba, Spain 2IFAPA, Centro Alameda del Obispo, 14080 Córdoba, Spain 3University of Córdoba, Spain [email protected] Dry pea (Pisum sativum) is the most widely grown grain legume in Europe and second-most in the world. However, diseases are a strong limitation to obtain reasonable and stables yields in pea. At Córdoba we started pea research 10 years ago, have centred our activities mainly in resistance against broomrape (Orobanche crenata) as this was considered the major constraint. After a huge search we found that resistance is very scarce, being able to identify only some levels of incomplete resistance in a few accessions of P. sativum and in wild Pisum that were successfully crossed with pea cultivars and introduced into a breeding programme that is underway. Resistance is of complex inheritance, so looked QTLs and approached the characterisation of the mechanisms of resistance involved. We identified also resistance to Aschochyta blight (Mycosphaerella pinodes) and to powdery mildew (Erysiphe pisi). Studies on the inheritance and identification of QTLs for M. pinodes resistance have been performed. A breeding program including these resistant accessions is also in progress. We have identified a new gene for resistance to E. pisi, called Er3. This gene from P. fulvum has been successfully introduced into pea varieties and we have identified SCARs markers linked to this gene that will facilitate the early selection of individuals carrying the gene in breeding programs. At present we are approaching the study of the molecular and biochemical mechanisms of response to M. pinodes and E. pisi in pea by micro-arrays, transcription factors and proteomics analysis. Even, when our major goal is to understand and exploit genetic resistance in breeding, we assume that existing levels of resistance might still be insufficient for an effective crop protection. Thus, we also try to complement resistance with other strategies of integrated control, including studies on activation of systemic acquired resistance and on the effect of fertilisation and of intercropping on disease.

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Incidence of viruses in the most important bean

varieties in Vojvodina

D. Petrović1, 1M. Ignjatov1, F. Bagi2, M. Milošević3, M. Vasić1 and M. Vujaković1

1Institute of Field and Vegetable Crops, Novi Sad, Serbia 2University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia 3Ministry of Agriculture, Forestry and Water Management, Belgrade, Serbia [email protected]

Legume production is becoming increasingly important in the province of Vojvodina. The bean varieties grown in Serbia are mostly of domestic origin and must be checked for the presence of harmful pathogens, including viruses. Pathogens causing viral diseases have not been sufficiently studied in the country to date. The present study tested eight bean varieties (Sremac, Dvadesetica, Belko, Zlatko, Ribničanin, Slavonski Zeleni, Tisa, and Domaća Populacija) for infection by the Bean common mosaic virus, Bean common mosaic necrosis virus, Bean yellow mosaic virus, Cucumber mosaic virus, and Alfalfa mosaic virus at several locations in the Vojvodina province. The said viruses are capable of causing great damage in bean production, which is compounded by the high incidence of their vectors (leaf aphids) in bean plants. The harmfulness of viral diseases varies and is dependent on the susceptibility of the cultivar, the stage of plant development at which the infection occurs, and environmental conditions affecting the activity of the vectors. In our study, virus identification in the cultivars was carried out by the DAS ELISA test using a commercial set of antiserums for BCMV, BCMNV, BYMV, CMV, and AMV according to the procedure recommended by the manufacturer, LOEWE Biochemica GmbH, Germany. The results of the ELISA test were read on a Multiskan Ascent microplate reader at a wavelength of 405 nm. An extinction coefficient value that was found to be double that of the negative control was considered a positive result. The results of the serological analysis showed that the largest number of infected plants was present in the cultivar Dvadesetica. As many as 72% of the plant samples of this variety were found to be infected by the viruses concerned. By virus, the infection percentages in Dvadesetica were as follows: BCMV – 32%, AMV – 28%, CMV – 8%, and a mixture of BCMV and AMV – 4%. The second and third most infected varieties were Zlatko (71%) and Domaća Populacija (70%), respectively. The smallest number of infected plants was recorded in the variety Sremac (16%). In the rest of the cultivars, the infection percentage ranged between 30 and 57%. The most commonly found virus was BCMV, while BYMV was not found in any of the cultivars. Continuous pathogen identification and crop control are needed in order to develop varieties tolerant or resistant to pathogens.

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Race identification of Pseudomonas syringae pv. glycinea

on commercial soybean varieties in Serbia

M. Ignjatov1, M. Vidić1, M. Milošević2, J. Balaž3, M. Vujaković1, D. Petrović1

1Institute of Field and Vegetable Crops, Novi Sad, Serbia 2Ministry of Agriculture, Forestry and Water Management, Belgrade, Serbia 3University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia [email protected] Bacterial blight, caused by the bacterium Pseudomonas syringae pv. glycinea (Coerper) Young et al., is one of the most economically important diseases of soybean. It has long been known that many soybean genotypes possess different levels of susceptability or resistance to this pathogen. The growing of susceptible soybean varieties is the main reason behind the high frequency and severity of bacterial blight outbreaks around the world, Serbia included. Data on soybean cultivars resistant to the disease are relatively scarce. The reason for this may lie in the fact that a number of different races of the pathogen have been identified over the last few decades, making it much more difficult to develop cultivars resistant to all the known races. So far, 12 physiological races of P. s. pv. glycinea have been detected in total (Gao Jie et al., 1998). In the present study, the pathogen was isolated from the leaves of the diseased plants, with the leaf samples having been collected from different soybean varieties in several locations in the province of Vojvodina. Pathogenicity of the obtained isolates including referent strain (National Collection of Plant Pathogenic Bacteria, United Kingdom-NCPPB 3318) was proved by inoculation soybean plants at cotyledon stage (Balkan variety) and by hypersensitive reaction (HR) on tobacco leaves. The racial identity of the isolates was determined using a differential set of soybean varieties (Acme, Chippewa, Flambeau, Harosoy, Lindarin, Merit i Norchief) according to Cross et al. (1966). Ten days after emergence, young soybean plants were inoculated with a bacterial suspension containing all the isolates tested. Inoculation was done by pressurized spraying using a hand-held atomizer. Inoculation was performed during the early development of the first trifoliate leaf, with the leaflets having reached ½ of their normal size (Phenophase V2). A bacterial suspension concentration of 108 cfu/ml was used for inoculation. The photoperiod in the greenhouse was 12 h light, 12 h dark. The first reading was made 72 h after inoculation, while the final evaluation was performed after 14 days. The reaction of the differential set of soybean varieties indicated a clear presence of susceptibility in all the cultivars, and all the strains used were determined to belong to race 4 of the pathogen. In their study of P. s. pv. glycinea isolates from soybean in Vojvodina, Balaž et al. (1990) also established the presence of race 4 in the country. According to Sinclair (1999), there are four dominant genes controlling resistance to P. s. pv. glycinea (Rpg1, Rpg2, Rpg3 and Rpg4). The resistance of soybean to this pathogen is also affected by a host of other factors within the plants, one being an increased production of ethylene that reduces the virulence of the bacterium. Based on this findings, it can be concluded that the bacterial blight of soybean must be continuosly monitored and studied in the future and that cultivars with reduced susceptability to P. s. pv. glycinea should be developed in order to improve soybean production in this regard. Gao Jie (1998): Journal of Jiling Agricultural University, 20, 10-12. Cross J. E., Kennedy B. W., Lambert J. W. and Cooper R. L. (1966) Plant Dis. Rep. 50:557-560. Sinclair J. B. (1999) In: Hartman G. L., Sinclair J. B., Rupe J. C. (ed) Compedium of soybean diseases. APS Press. USA, 5-10. Balaž J., Arsenijević M. and Vidić M. (1990) Zaštita bilja, 41, 4, 194, 423-429.

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Pod growth temperature affects the granular structure of

starch from pea (Pisum sativum L.) seeds

N. A. Roder, T. Y. Bogracheva, D. A. Jones and C. L. Hedley

John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, United Kingdom [email protected] Pods from three pea (Pisum sativum L.) mutants (r-f, rug5-a and lam-c) with seeds that differ in starch content and composition, plus pods from the non-mutant wild type (WT), were grown to maturity at 10ºC and 30ºC, while maintaining the plants and ‘control’ pods at 16/12ºC (day/night) conditions. After starch extraction, the effect of the growing environment on starch structure was determined at a molecular level, by measuring the proportion of amylose/amylopectin and the distribution of amylopectin chains with different degrees of polymerisation (DP) and at a supramolecular level, by determining the proportion of double helices and crystallinity and the polymorph type and content. It was found that the proportion of amylose decreased with increasing growth temperature for the WT, r-f, and rug5-a lines, but increased in the lam-c line. There was no significant effect of temperature for any of the lines on the amylopectin chain-length distributions. The major effect for all the lines was a large increase in the A-polymorph content of starches from seeds grown at 30ºC, which was linked to a similar increases in the gelatinisation peak temperatures. A small, but significant, increase was found in starch crystallinity with increased growth temperature, but there was no significant effect on the proportion of double helices. It is proposed that the molecular structure of starch is mainly determined by the genotype. However, the supramolecular structure, in particular the type and proportion of polymorphs present, was affected by the thermodynamic environment within which the starch develops.

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Grain yield components in winter genotypes of faba bean

(Vicia faba L.)

M. Vasić1, A. Mikić1, V. Mihailović1, B. Ćupina2, Đ. Krstić2 and G. Duc3

1Institute of Field and Vegetable Crops, Novi Sad, Serbia 2University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia 3Institut National de la Recherche Agronomique, Unité Mixte de Recherche en Génétique et Ecophysiologie des Légumineuses à Graines, Dijon, France [email protected] Faba bean (Vicia faba L.) is usually grown as spring crop in Serbia with its continental climatic conditions (3). Recently the first Serbian breeding programme on the development of winter lines of both food and feed faba bean has been initiated, being directed towards sufficient winter hardiness and yield of grain at a level of widely distributed spring cultivars (2). The development of winter cultivars of faba bean is based upon the breeding for freezing tolerance and is aimed towards enlarging the cultivation zone of faba bean (1). A small-plot trial was carried out from the autumn of 2006 to the spring 2008 at the Experimental Field of the Institute of Field and Vegetable Crops at Rimski Šančevi on a carbonated chernozem soil. It included 11 faba bean genotypes, namely three Serbian food faba bean local landraces (Ashtar, Crveni Kamnica and Višnjićevo), three Serbian feed faba bean local landraces (PP 1, PP 2 and PP5), two Serbian feed faba bean cultivars (Gema and Šarac), two French faba bean cultivars (Diva and Irena) and one German faba bean population (Göttingen Winter Bean Population). All cultivars were sown in both years in early October, with an average crop density of 45 viable seeds m-2. There were monitored plant height (cm), first fertile node height (cm), number of nodes (plant-1), ordinal number of the first fertile node, number of fertile nodes (plant-1), number of pods (plant-1), number of grains (plant-1), thousand grains mass (g) and grain yield (g plant-1), as well as the ratio between the number of plants before and after the winter. The highest grain yield was in Šarac (16.49 g plant-1), while Ashtar had the highest ratio between the number of plants before and after the winter (38 m-2). (1) Duc G. (1997) Field Crops Research, 53, 99-109. (2) Mihailović V., Mikić A., Ćupina B., Krstić Đ., Erić P., Milić D. and Vasić M. (2006) International Workshop on Faba Bean Breeding and Agronomy, Córdoba, Spain, 25-27 October 2006, 93-95. (3) Mihailović V., Mikić A., Ćupina B., Vasić M. and Erić P. (2007) A Periodical of Scientific Research on Field and Vegetable Crops, 43, 255-261.

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Challenges in breeding subtropical legumes for temperate

regions

V. Mihailović1, A. Mikić1, B. Ćupina2 and M. Vasić1

1Institute of Field and Vegetable Crops, Novi Sad, Serbia 2University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia [email protected] Pigeonpea (Cajanus cajan (L.) Millsp.), hyacinth bean, (Lablab purpureus (L.) Sweet) and cowpea ( Vigna unguiculata (L.) Walp.) originate from the common African centre of diversity. Being typical tropical and subtropical legumes, they are not familiar in Serbia, placed in the central and northern parts of the Balkan Peninsula and with a temperate climate and geographical position of approximately between 42° and 46° N. It can be considered that there are no written records of any serious attempt on cultivating either pigeonpea or hyacinth bean in Serbia, while cowpea is not completely unknown in the country. A long-term evaluation of forage and grain yields in these three species has shown that all of them have considerable potential for both forage and grain production. The pigeonpea cultivar Quest had the highest yields of green forage and forage dry matter (52.8 t ha-1 and 14.8 t ha-1), the hyacinth bean genotype NI 77R produced 34.1 t ha-1 of green forage and 9.5 t ha-1 of forage dry matter and Xincharo was the most yielding of all cowpea accessions, producing 45.9 t ha-1 of green forage and 12.9 t ha-1 of forage dry matter (2). The average grain yield varied from 719 kg ha-1 in ICPL 88020 to 2053 kg ha-1 in Quest in pigeonpea and from 57 kg ha-1 in Xincharo to 3460 kg ha-1 in NI 147 in cowpea, while the average grain yield in hyacinth bean NI 77R was 630 kg ha-1 (1). The most important characteristic for the breeding programme of these species in Serbia is photoperiod reaction. Due to inadequate day length, several genotypes of all these three species, including others such as Bengal bean (Mucuna pruriens (L.) DC.), remain constantly in a vegetative stage, producing extremely abundant biomass of more than 50 t ha-1, mostly without flowers and absolutely with no pods and seeds, until the end of the whole growing season. By that reason, the selection of the genotypes able to produce seeds is of the highest importance and only through this characteristic it is possible to adjust others that are also desirable, such as earliness and high and quality yields of both forage and grain. Acknowledgements: Ms. Edna Matsuda of Com. Ind. Matsuda Imp. e Exp. Ltda, Álvares Machado, Brasil. (1) Mihailović V., Mikić A., Vasiljević S., Milić D. Vasić M. and Ćupina B. (2005) Proceedings of the 1st International Edible Legume Conference in conjunction with the IVth World Cowpea Congress, Durban, South Africa, 17-21 April 2005, CD Rom. (2) Mihailović V., Mikić A., Vasiljević S., Milić D., Ćupina B., Krstić Ð. and Ilić O. (2006) Grassland Science in Europe, 11, 306-308.

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White lupin (Lupinus albus L.) landraces and the breeding

for tolerance to alkaline soil reaction

M. Vishnyakova1 and A. Mikić2 1N. I. Vavilov Research Institute of Plant Industry (VIR), St. Petersburg, Russia 2Institute of Field and Vegetable Crops, Novi Sad, Serbia [email protected] White lupin (L. albus L.) is generally considered unsuitable for the cultivation on alkaline soils such as chernozem, present in many countries of the eastern part of the Europe. A high soil pH, together with presence of CaCO3, may cause a lack of iron in the soil and lead to both chlorosis and necrosis in white lupin plants (2). A small-plot trial has been carried out in 2007-2008 at the Experimental Field of the Institute for Field and Vegetable Crops at Rimski Šančevi at the carbonated chernozem with pH =7.92. The trial included five white lupin landraces from the VIR collection: K-507 and K-509 from Egypt, K-490 and K-494 from Ethiopia and K-305 from Israel collected in 1926-27 by N.I.Vavilov. Previously they had been evaluated at VIR’s Ukrainian branch for main agronomic and biochemical traits. All accessions had high protein (38.4% - 45.3%) and oil content (8.6-10.2%) in the seeds, two genotypes (K-494 and K-507) revealed high tolerance to fusarium. The sample K-494 had the highest yield 292-346 g/m2 at the density 22 plants/ m-2. Period of maturation differed between the landraces from 116 up to 150 days (3). At Rimski Šančevi, the landraces were sown in the first half of March, 50 viable seeds m-2, a plot size of 5 m-2 and three replicates, harvested at the stage of full maturity of the second-order grains and before the shattering of the first-order pods. Grain yield components with the emphasis upon the distribution by orders and other agronomic characteristics have been evaluated. All accessions had greater number of second-order grains per plant in comparison to the number of first-order grains. In the landraces K-507, K-494 and K-490, the first-order grains were the largest, while the third-order grains were the smallest. On the other hand, in the landrace K-509 the second-order grains were the largest and the third-order grains were the smallest, while in the landrace K-305 the first-order grains were the largest and the second-order grains were the smallest. In average, thousand grains mass varied between 180 g in the third-order grains of the landrace K-490 and 387 g in the first-order grains of the landrace K-507. The landrace K-494 had the highest average yield per area unit (7395 kg ha-1). Unlike the preliminary results in the same conditions (1), three of the five examined landraces had the second order as more productive than the first one, where the landrace K-305 had the highest second-order grain yield proportion (0.58). The obtained results may be regarded as rather promising for the future white lupin breeding programmes for alkaline soil types. (1) Ćupina B., Mikić A., Mihailović V., Krstić Đ. and Erić P. (2007) Book of Abstracts of the 6th European Conference on Grain Legumes, Lisbon, Portugal, 12-16 November 2007, 117. (2) Mihailović V., Ćupina B., Hill G. D., Mikić A., Święcicki W., Jones R. and Eickmeyer F. (2006) Proceedings of the 11th International Lupin Conference, Guadalajara, Mexico, 4-9 May 2005, 99-101. (3) Reference VIR catalogue, № 496. (1989). Lupinus albus L. VIR. St. Petersburg, Russia, 50.

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139

Index of authors

Abdel-Galil M. M. 106, 108 Abdelly C. 78 Adesoye A. I. 76 Adhikari B. N. 64, 90 Akhtemova G. 112 Angelova S. 30 Annicchiarico P. 20 Bagi F. 128 Balaž J. 130 Balešević-Tubić S. 18, 104 Baral B. R. 64 Barilli E. 126 Bastola B. S. 64 Bett K. 70 Bing D. 70 Biswas B. 120 Bogracheva T. Y. 132 Borisov A. 112 Bugayov V. D. 94 Carroll B. 120 Castellejo M. A. 126 Chan P. K. 120 Chebotar V. 112 Čupić T. 58, 88 Ćupina B. 32, 34, 36, 40, 44, 134, 136 Curto M. 126 Darai R. 64, 90 Delgado M.-J. 78 Delić D. 122 Đorđević V. 18, 44, 82, 104 Đukić V. 18, 104 Drezner G. 88 Duc G. 134 Ellis T. H. N. 14 Emese A. 76 Erić P. 36 Fatnassi N.78 Ferguson B. 120 Fernández-Aparicio M. 126 Fondevilla S. 126 Gabrovská D. 38 Gantner R. 58, 88 Ghaouti L. 48 Gharti D. B. 90 Gianinazzi-Pearson V. 112 Gresshoff P. M. 120 Grishina O. 112 Hamed N. M. 106, 108 Hauptvogel P. 32

Hauptvogel R. 40 Hedley C. L. 132 Helmy A. A. 108 Hirani T. 120 Horres R. 78 Ignjatov M. 128, 130 Imtiaz M. 24 Indrasumunar A. 120 Ivanović M. 86 Ivanyuk S. V. 62 Jarak M. 114 Jiang Q. 120 Jones D. A. 132 Jovićević D. 34 Kahl G. 78 Karagić Đ. 56, 98 Katić S. 56, 96, 98 Kavar T. 118 Kazakov A. 112 Kereszt A. 120 Khan F. 78 Kidrič M. 118 Kinkema M. 120 Kirby G. 100 Kolisnyk S. I. 62 Korakhashvili A. 124 Kovačević B. 22 Krasheninnikova A. 112 Krstić Đ. 32, 34, 36, 40, 134 Kumar Y. 72 Kuzmanović Đ. 122 Li D. 120 Lin M.-H. 120 Lin Y.-H. 120 Ljuština M. 42, 44 Lugić Z. 54, 86 Maksimov A. M. 94 Maksimović S. 122 Maras M. 118 Marinković J. 114 Matić R. 100 McPhee K. 116 Meglič V. 118 Mihailović V. 32, 34, 36, 40, 44, 56, 98, 134, 136 Mikić A. 32, 34, 36, 40, 42, 44, 56, 96, 98, 104, 134, 136, 138 Miladinović J. 18, 82, 104 Miličić B. 122

Milić D. 34, 56, 96 Milošević M. 128, 130 Mishra S. K. 72 Miyagi M. 120 Miyahara A. 120 Mladenović-Drinić S. 74 Molina C. 78 Moloshonok A. 112 Moral A. 126 Nagel S. 100 Naumkina T. 112 Nemankin T. 112 Neupane R. K. 64 Nontachaiyapoom S. 120 Orlović S. 22 Ouhrabková J. 38 Ovchinnikova E. 112 Pataki I. 98 Paulíčková I. 38 Pérez-de-Luque A. 126 Perić V. 74, 92 Petrović D. 128, 130 Petrychenko V. F. 62 Pilipović A. 22 Pokhrel D. N. 90 Popović S. 58, 88 Prats E. 126 Radović J. 54 Rasulić N. 122 Reid D. 120 Roder N. A. 132 Rubiales 126 Rysová J. 38 Sarker A. 72, 90 Sarukhanyan N. 60 Sass O. 48 Scotti C. 20 Sharma B. 16, 72 Shtark O. 112 Sillero J. C. 126 Smith K. 100 Smýkal P. 28, 80 Sokolović D. 54 Srebrić M. 74, 92 Stajković O. 122 Steinhauer D. 78 Stjepanović M. 58, 88 Stoddard F. L. 50 Štrbanović R. 54 Šurlan-Momirović G. 86 Šuštar-Vozlič J. 118

Tar’an B. 70 Taški-Ajduković K. 82 Tikhonovich I. 112 Torres A. M. 126 Tyagi M. C. 72 Ubayasena L. 70 Vanyan A. 60 Vasić M. 52, 114, 128, 134, 136 Vasić T. 54 Vasilchikov A. 112 Vasiljević S. 44, 56, 86, 96 Vidić M. 82, 130 Vishnyakova M. 138 Vujaković M. 82, 128, 130 Vymyslický T. 38 Wang X. 116 Warkentin T. 70 Winter P. 68, 78 Yadav N. K. 90 Young I. 100 Zadorozhnyi V. S. 94 Zdravković M. 52 Zhukov V. 112

List of participants

ADESOYE Adenubi University of Ibadan Ibadan Nigeria [email protected] ANGELOVA Siyka Institute of Plant Genetic Resources Sadovo Bulgaria [email protected] ANNICCHIARICO Paolo Centro di Ricerca per le Produzioni Foraggere e Lattiero-Casearie Lodi Italy [email protected] BOCHARD Anne-Marie Limagrain Verneuil Holding Riom France [email protected] BORISOV Alexey All-Russia Research Institute for Agricultural Microbiology St. Petersburg Russia [email protected] BUGAYOV Vasyl Feed Research Institute of the Ukrainian Academy of Agrarian Science Vinnica Ukraine [email protected] ĆUPINA Branko Faculty of Agriculture Novi Sad Serbia [email protected]

DARAI Rajendra Nepal Agricultural Research Council Nepalunj Nepal [email protected] DELIĆ Dušica Institute of Soil Science Belgrade Serbia [email protected] ĐORĐEVIĆ Vuk Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] DOSTALOVÁ R. Agritec Plant Research Ltd. Šumperk Czech Republic [email protected] ELLIS T. H. Noel John Innes Centre Norwich UK [email protected] GANTNER Ranko Faculty of Agriculture Osijek Croatia [email protected] GHAOUTI Lamiae Norddeutsche Pflanzenzucht Hans-Georg Lembke KG Holtsee Germany [email protected] HOCHMAN M. Agritec Plant Research Ltd. Šumperk Czech Republic [email protected]

HYBL Miroslav Agritec Plant Research Ltd. Šumperk Czech Republic [email protected] IGNJATOV Maja Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] IVANYUK Sergey Feed Research Institute of the Ukrainian Academy of Agrarian Science Vinnica Ukraine [email protected] KARAGIĆ Đura Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] KATIĆ Slobodan Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] KOLISNYK Sergey Feed Research Institute of the Ukrainian Academy of Agrarian Science Vinnica Ukraine [email protected] KOVAČEVIĆ Branislav Institute for Lowland Forestry and Environment Novi Sad Serbia branek@uns. ns.ac.yu KRSTIĆ Đorđe Faculty of Agriculture Novi Sad Serbia [email protected]

LUGIĆ Zoran Institute for Forage Crops Kruševac Serbia [email protected] LJUŠTINA Marija Faculty of Philosophy Belgrade Serbia [email protected] MARAS Marko Agricultural Institute of Slovenia Ljubljana Slovenia [email protected] MARINKOVIĆ Jelena Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] MARKOVÁ Helena Research Institute for Fodder Crops Troubsko Czech Republic [email protected] McPHEE Kevin North Dakota State University Fargo USA [email protected] MIHAILOVIĆ Vojislav Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] MIKIĆ Aleksandar Institute of Field and Vegetable Crops Novi Sad Serbia [email protected]

MILADINOVIĆ Jegor Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] MILIĆ Dragan Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] PERIĆ Vesna Maize Research Institute Zemun Polje Belgrade-Zemun Serbia [email protected] PETROVIĆ Dragana Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] RADOVIĆ Jasmina Institute for Forage Crops Kruševac Serbia [email protected] RUBIALES Diego Institute for Sustainable Agriculture Córdoba Spain [email protected] SCHNEIDER Anne GL-TTP Paris France [email protected] SHARMA Balram Lentil Research Association New Delhi India [email protected]

SMÝKAL Petr Agritec Plant Research Ltd. Šumperk Czech Republic [email protected] SREBRIĆ Mirjana Maize Research Institute Zemun Polje Belgrade-Zemun Serbia [email protected] STODDARD Fred University of Helsinki Helsinki Finland [email protected] TAŠKI-AJDUKOVIĆ Ksenija Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] VANYAN Armen Green Lane Agricultural Assistance NGO Yerevan Armenia [email protected] VASIĆ Mirjana Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] VASILJEVIĆ Sanja Institute of Field and Vegetable Crops Novi Sad Serbia [email protected] VYMYSLICKÝ Tomáš Research Institute for Fodder Crops Troubsko Czech Republic [email protected]

WARKENTIN Tom University of Saskatchewan Saskatoon Canada [email protected] WINTER Peter GenXPro GmbH Frankfurt-am-Main Germany [email protected]

ZADOROZHNYI Viktor Feed Research Institute of the Ukrainian Academy of Agrarian Science Vinnica Ukraine [email protected]

Documents

Book of Abstracts - Institut za ratarstvo i povrtarstvo, Novi Sad...(GenXPro GmbH, Frankfurt, Germany) Local Organising Committee Branko Ćupina (Faculty of Agriculture, Novi Sad,