Abstracts 27th ISTA Congress Seed Symposium Budapest, Hungary May 17th – 19th, 2004

Compiled by: The International Seed Testing Association (ISTA) P.O.BOX 308 8303 Bassersdorf, CH-Switzerland

ISBN: 3-906549-44-6 Edition 2004: 500 copies

Copyright©2004 by the International Seed Testing Association (ISTA)

All rights reserved. No part of this publication may be reproduced, stored in retrieval system or transmitted in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, without the prior permission in writing of ISTA.

Budapest, Hungary, May 17th – 19th, 2004

Preface

Dear Seed Symposium Participants

Welcome to Budapest at the Seed Symposium of the 27th Congress of the International Seed Testing Association (ISTA).

On behalf of the ISTA, I have the honor to present you the Programme and the Abstracts of the ISTA Seed Symposium.

The ISTA Seed Symposium is probably ‘the largest international gathering of applied seed scientists and is an unique forum for the interchange of ideas between seed scientists and technicians’.

‘Towards the Future in Seed Production, Evaluation and Improvement’, is the theme of the Seed Symposium. Scientists from 45 countries worldwide submitted over 290 high quality papers on various interesting topics and for the seven sessions of the Seed Symposium.

Six of these papers are presented as oral presentations in each of the sessions. Three key- note speeches will complete the programme. A detailed programme and all abstracts of papers submitted for the Seed Symposium of the 27th ISTA Congress, held in Budapest, May 17th – 19th, 2004, can be found in this abstract booklet.

The abstracts are thematically arranged according to session and listed within each session alphabetically according to the authors names. Readers desiring a follow-up should contact the authors directly.

I hope all of you will find the Seed Symposium informative and interesting. I sincerely believe that it will provide an useful forum for the exchange of ideas and the latest results in seed testing topics, as well as an opportunity to meet experts from all over the world.

Please enjoy the ISTA Seed Symposium.

Dr Alison Powell Symposium Convenor

1 27th ISTA Congress – Seed Symposium

Contents

Programme 3

Opening Ceremony 6

Oral Presentations 6 Keynote byZoltan Syposs 6 Session 1: Application of Advanced Technologies 6 Session 2: Organic and Conventional Seed Production 10 Keynote by Murry Hill 13 Session 3. Viability and Vigour: Evaluation and Impact 13 Session 4: Seed Systems in Emerging and Developing Economics 16 Session 5: Seed Lot Hygiene 19 Keynote by Gerry Saddler 23 Session 6: Seed Improvement 23 Session 7: Physiological Basis of Seed Quality 26

Poster Sessions 29 Session 1: Application of Advanced Technologies 29 Session 2: Organic and Conventional Seed Production 45 Session 3: Viability and Vigour: Evaluation and Impact 58 Session 4: Seed Systems in Emerging and Developing Economics 96 Session 5: Seed Lot Hygiene 102 Session 6: Seed Improvement 119 Session 7: Physiological Basis of Seed Quality 136

2 Budapest, Hungary, May 17th – 19th, 2004

ISTA Seed Symposium 2004

‘Towards the Future in Seed Production, Evaluation and Improvement’

Programme

Time Monday, May 17, 2004

OPENING CEREMONY - Official Address by the Representative of the Hungarian Minister of Agriculture & Regional Development. 08:30 – 10:00 - Opening by the ISTA President - Welcoming Address by the ISTA Secretary General - Welcoming Address by the Organisers - Greetings from other International Organisation

10:00 – 10:30 COFFEE BREAK

KEYNOTE – Future developments in the seed industry in Eastern Europe, by 10:30 – 11:00 Zoltan Syposs, Hungary

SESSION 1 – Application of Advanced Technologies Chaired by Enrico Noli, Italy 11:00 – 11:20 - ‘A computerised key for seed identification’ by M.L. Gupta, D.L. George & B.B. Basnet, Australia 11:20 – 11:40 - ‘Use of near infrared reflectance spectroscopy to identify seeds of noxious weeds, forage legume seeds and contamination’ by W. Hugo & P. Dominguez, Uruguay 11:40 – 12:00 - ‘Biochemical characterization of white onion ecotypes (Allium cepa L.) through HPLC analysis of seed storage proteins’ by G. Mennella, V. Onofaro Sanaja’, A. D’Alessandro & M. Milone, Italy

12:00 – 13:00 LUNCH

SESSION 1 – Application of Advanced Technologies (cont.) 13:00 – 13:20 - ‘Commercial sunflower germplasm identification and characterization using SSR’ by A. Vicario, A. Loray, N. Paniego & E. Hopp, Argentina 13:20 – 13:40 - ‘Development of microsatellite markers for the identification of Brazilian Coffe Arabica Cultivars’ by E.S.N. Vieira, E.V.R. von Pinho, D.G. Esselink, M.G.G.C. Vieira & B. Vosma, Brazil & The Netherlands 13:40 – 14:00 - ‘Testing for adventitious presence of transgenic material in conventional seed lots using quantitative laboratory methods: a new statistical approach and its implementation’ by J.-L. Laffont, K.M. Remund, D.L. Wright & R.D. Simpson, France & USA

SESSION 2 – Organic & Conventional Seed Production Chaired by José de Barros França Neto, Brazil 14:00 – 14:20 - ‘Organic seed treatment to control common bunt (Tilletia tritici) in wheat’ by A. Borgen, Denmark 14:20 – 14:40 - ‘A comparative study of the germination characteristics of wild flower seeds in commercial seed production’ by M. Khajeh-Hosseini, A.A. Powell & G.K. Laverack, United Kingdom 14:40 – 15:00 - ‘Varietal characterization and genetic purity assessment of castor (Ricinus communis L.) genotypes’ by R. Ankaiah, N. Manohar Reddy K. Keshavulu & P. Sambasiva Rao & B. Muralimohan Reddy, India

15:00 – 15:30 COFFEE BREAK

SESSION 2 – Organic & Conventional Seed Production (cont.) 15:30 – 15:50 - ‘The effect of glyphosate treatment on the germination potential of barley seed’ by G. McLaren & R. Don, United Kingdom 15:50 – 16:10 - ‘Challenges for obtaining high quality organic seeds’ by S.P.C. Groot & W.J. van der Burg, The Netherlands 16:10 – 16:30 - ‘Effect of desiccation on some quality characteristics’ by M. Rajic, B. Marinković, V. Miklič & L. Panković, Serbia and Montenegro

3 27th ISTA Congress – Seed Symposium

SESSION 3 – Viability and Vigour: Evaluation and Impact Chaired by Joël Léchappé, France 16:30 – 16:50 - ‘Relationship between standard and cold germination tests in supersweet sweetcorn’ by D.L. George, M.L. Gupta & I.G.M.A. Parwata, Australia 16:50 – 17:10 - ‘Assessment of reliability of germination papers in environmental stress research’ by M. Khajeh-Hosseini, A.A. Powell & I. Bingham, United Kingdom 17:10 – 17:30 - ‘Assessment of viability of bread wheat (Triticum aestivum L.) and oats (Avena sativa L.) germplasm samples stored over 30 years in cold store’ by L. Holly, R. Bócsó, A. Juhász & I. Már, Hungary

Tuesday, May 18, 2004

KEYNOTE – Development of education and training in seed science and 08:30 – 09:00 technology, by Murray Hill, New Zealand

SESSION 3 – Viability and Vigour: Evaluation and Impact (cont.) 09:00 – 09:20 - ‘Vigour testing: towards an extended use of the conductivity test’ by M.-H. Wagner, A. Preveaux, E. Moizan, M. Beaulaton & S. Ducournau, France 09:20 – 09:40 - ‘Vigour tests for predicting seedling emergence of aubergine (Solanum melongena L.) seed lots’ by I. Demir, S.Ermis, G. Okcu & S. Matthews, Turkey & United Kingdom 09:40 – 10:00 - ‘Performance of osmoprimed seed of germplasm of desi and kabuli chickpeas under laboratory and field conditions’ by S.J. Singh & K. Singh, India

10:00 – 10:30 COFFEE BREAK

SESSION 4 – Seed Systems in Emerging and Developing Economies Chaired by Grete Tarp, Denmark 10:30 – 10:50 - ‘High quality rice seed production - Santa Catarina model’ by R. Knoblauch, Brazil 10:50 – 11:10 - ‘Applied tree seed technology in Brazilian Atlantic forest’ by Fatima C.M. Piña-Rodrigues & E.S. Nogueira, Brazil 11:10 – 11:30 - ‘Seed sector development and seed policies implemented in Turkey’ by B. Bozkurt, K. Yalvec & O.F. Bal, Turkey

11:30 – 12:30 POSTER SESSION 1

12:30 – 13:30 LUNCH

SESSION 4 – Seed Systems in Emerging and Developing Economies (cont.) 13:30 – 13:50 - ‘Potential of on-farm practices for improving rice seed quality, seed health and crop production’ by S.B. Mathur, C.N. Mortensen, M.H. Talukder & R.B. Mabagala, Denmark, Bangladesh & Tanzania 13:50 – 14:10 - ‘Seed programme development in a transition economy – the experience of Vietnam’ by T.D.N. Dung & M. Turner, Vietnam 14:10 – 14:30 - ‘Seed sector development in South Western Nigeria: farmers' utilization of improved seed for crop production' by I.O. Daniel & J.A. Adetumbi, Nigeria

SESSION 5 – Seed Lot Hygiene Chaired by Ákos Mesterhazy, Hungary 14:30 – 14:50 - ‘Black Rot eradication treatments on Brassica: efficacy and seed quality investigations’ by A.G. Taylor, J.D. Klein & R.H. Morrison, United States 14:50 – 15:10 - ‘Factors affecting the occurrence of Fusarium spp. in cereal seeds in Norway’ by G. Brodal & O. Elen, Norway 15:10 – 15:30 - ‘Seed borne barley stripe mosaic virus in Egypt: incidence, effect of virus and seed-transmisiblity’ by S. Zein & A. Aboul-Ata, Egypt

15:30 – 16.00 COFFEE BREAK

SESSION 5 – Seed Lot Hygiene (cont.) 16:00 – 16:20 - ‘Mycological aspects of onion seed priming’ by H. Dorna, K. Tylkowska, R. Marcinek, D. Szopinska & W. Yahong, Poland & China 16:20 – 16:40 - ‘Health status of sweet corn seeds in China and its impact on seedling emergence’ by Z. Guangwu, Z. Guozhen & W. Jianhua, China 16:40 – 17:00 - ‘Using seed health testing results to make balanced seed management decisions’ by V. Cockerell & M. McNeil, United Kingdom 17:00 – 17:20 - ‘Disease free seed production of wheat in Punjab (India): achievements and constraints’ by R.C. Sharma, I. Sharma & J.S. Samra, India

4 Budapest, Hungary, May 17th – 19th, 2004

Wednesday, May 19, 2004

08:30 – 09:00 KEYNOTE – Molecular methods and the future seed testing, by G. Saddler, United Kingdom

SESSION 6 – Seed Improvement Chaired by Hugh W Pritchard, United Kingdom 09:00 – 09:20 - ‘Seed testing and the effect of insecticidal active ingredients on emergence of hybrid maize seed’ by A. Jonitz & N. Leist, Germany 09:20 – 09:40 - ‘Effects of sand priming on germination, physiological changes and field performance in direct-sown rice (Oryza sativa L.)’ by J. Hu, Z.Y. Zhu, W.J. 09:40 – 10:00 Song, J.C. Wang & R. Naganagouda, China - ‘Effect of salt priming on seedling emergence and growth of watermelon seeds during development’ by I. Demir & K. Mavi, Turkey

10:00 – 10:30 COFFEE BREAK

SESSION 6 – Seed Improvement (cont.) 10:30 – 10:50 - ‘Sulphuric acid scarification effects on Brachiaria brizantha, B. humidicola and Panicum maximum seed dormancy releasing’ by R. Usberti & L. Martins, Brazil 10:50 – 11:10 - ‘Effects of different polymer coating materials and applications on the storage life and ageing of onion (Allium cepa L.cv. Aki) seeds: I. Assessment of appropriate polymers and dozes’ by S. Kavak & B. Eser, Turkey 11:10 –11:30 - ‘The use of rapid ageing and controlled deterioration to evaluate iodine vapour treatments to improve seed storage potential’ by N. De Atrip, S. Matthews & A.A. Powell, United Kingdom

11:30 – 12:30 POSTER SESSION 2

12:30 – 13:30 LUNCH

SESSION 7 – Physiological Basis of Seed Quality Chaired by Françoise Corbineau, France 13:30 – 13:50 - ‘Effect of high temperature stress during soybean seed development on germination and vigor’ by D.M. TeKrony, D.B. Egli & J. Spears, USA 13:50 – 14:10 - 'Genetic dissection of maize response to a defoliation treatment during maturation inducing tolerance to cold at germination' E. Frascaroli, E. Casarini & S. Conti, Italy 14:10 – 14:30 - ‘Lipid peroxidation and activity of superoxide dismutase associated with natural aging of oil maize seed’ by S. Balešević-Tubić, M. Tatić, J. Miladinović, Đ. Malenčić,, Serbia and Montenegro 14:30 – 14:50 - ‘Seed Longevity Chart and Modeling to Predict viability during open storage’ by C. Andreoli, Brazil

14:50 – 15:20 COFFEE BREAK

SESSION 7 – Physiological Basis of Seed Quality (cont.) 15:20 – 15:40 - ‘The effect of different post-harvest drying methods on seed quality from green (immature) and red (mature) berries of woody nightshade (Solanum dulcamara L.)’ by J. Coneybeer, J. Adams & R.J. Probert, United Kingdom 15:40 – 16:00 - ‘100-seed test for desiccation tolerance and germination: a case study on eight tropical palm species’ by C.B. Wood, S. Hodges, H.J. Vautier & H.W. Pritchard, United Kingdom

16:00 – 17:00 SYMPOSIUM CONCLUSION

5 27th ISTA Congress – Seed Symposium

Opening Ceremony

Keynote – Future Development in Seed Industry in Eastern Europe By Zoltan Syposs, Hungary

1. Seed industry in CEE after 1989 • Reduction of Eastern export • Disappearance of production and processing of some crops • Reduction of the state-owned seed processing plants • Focus on the single cross hybrids • Re-orientation of the exports • Emphasis made on the quality • Privatization processes finished 2. Re-orientation of seed production • Large farm structure with irrigation possibilities • Human competence • Cost competitiveness • Internationally recognized certification work by OMMI 3. Changes and new demands • A new business environment –Food Processing Industry and supermarkets 4. Increasing importance of foreign ownership in food industry • Increasing activity of dedicated multinational seed companies 5. Syngenta investments in Hungary A vegetable breeding and testing station in Ócsa A maize seed plant in Mezőtúr 6. EU Accession – a great step forward • Access to all varieties of EU Catalogue • New materials introduced earlier to the new markets • Export opportunities improving for growers 7. Future of Seed companies in the EU • Globalization: an opportunity for the market drivers and a threat for the small players • Expanding breeding and screening activities towards East Expansion profiting from the present infrastructure • Increased co-operation of national seed associations under the umbrella of ESA • Vegetable production follows the trends of Western Europe • Moving production areas 8. Objectives and Challenges • Keep the focus of agriculture within the EU • Strengthen relationship between major agricultural countries

SESSION 1 – Application of Advanced Technologies Chaired by Enrico Noli, Italy

A COMPUTERISED KEY FOR SEED IDENTIFICATION GUPTA, M.L., GEORGE, D.L., & BASNET, B.B.

School of Agronomy and Horticulture, University of Queensland, Gatton, Qld, 4343 Australia, [email protected]

Australian Quarantine and Inspection Service (AQIS) regulates the quarantine requirements regarding seed imports in Australia. There are a number of seed testing laboratories in Australia which are currently servicing AQIS needs to prevent the entry of prohibited and 6 Budapest, Hungary, May 17th – 19th, 2004 restricted seeds. Current practices of identifying seeds by comparing an unknown seed with samples of known seeds or photographs of seeds are time consuming, costly and inefficient. A Seed Identification Key using a computerised database has been developed to identify prohibited and restricted seeds. There are currently 78 prohibited and 47 restricted seeds in the database. Lucid software was used to develop the Key because of its versatility in handling both text and image data. A total of 21 externally visible seed characters were identified as most suitable for development of the Key. Explanatory images and notes are attached to the character states to assist the user in correct selection of the state. The Key will be helpful to quarantine officers as well as seed analysts working in seed testing laboratories throughout Australia. It can also be used as an educational tool by agricultural scientists, students and others interested in seeds.

USE OF NEAR INFRARED REFLECTANCE SPECTROSCOPY TO IDENTIFY SEEDS OF NOXIOUS WEEDS, FORAGE LEGUME SEEDS AND CONTAMINATION HUGO1, W. & DOMINGUEZ2, P.

1National Agricultural Research Institute (INIA), Head of Seed Unit and Gene Bank, La Estanzuela. Ruta 50 Km 11. Colonia – Uruguay, [email protected] 2Advanced student (thesis), Department of Botany, Facultry of Agronomy, Montevideo, Uruguay, Avda. Eugenio Garzón 780. Montevideo-Uruguay, [email protected]

Near infrared reflectance spectroscopy (NIRS) was used to identify seeds of forage legumes species and noxious weeds species. It was also used to measure contamination of two noxious weed seeds: Rumex conglomeratus and Silene gallica in Trifolium repens. Seeds of weed species were collected by hand and threshed simulating the mechanical damage that they receive in a commercial seed lot. Seed of forage legume were obtained from basic seed lots. All samples were purified by hand to 99.9% purity. A total of 15 species of weeds and 5 forage legumes were analysed using a NIRSystems monochromator model 6500, Silver Spring, MD, USA. Samples were scanned from 400 to 2500nm in reflectance mode. Data was stored to build a data base of spectroscopies, then cross validated. Finally the data base was externally validated by asking the reflectance reader to classify new unkown (to the apparatus) samples. A good identification by Nirs was found. Some species with similar seeds (i.e. Ammi viznaga – Ammi majus) were separately identified. Some species presented greater difficulty and higher standard error of prediction (Rumex pulcher, Medicago sativa, Plantago myosurus). To measure concentration of weeds, seeds of Trifolium repens were obtained from a basic seed lot with 99.7% purity and then contaminated with different concentrations of Rumex conglomeratus and Silene gallica (0- 0,3- 0,5- 1- 1,5- 2- 3 and 5%). Four replications were used for building the database of reflectances, plus one “unknown” sample for validation. Two predictions equations using MPLS regression technique were developed. Calibration for contamination of R. conglomeratus showed a standard error of calibration (SEC) of 0,596, standard error of prediction 1,28 and coefficient of multiple determination (R2) of 0,686. For S. Gallica the numbers were: standard error of calibration (SEC) of 0,047, standard error of prediction 0,49 and coefficient of multiple determination (R2) 0,924 These results showed a different ability by Nirs reader to identify weed seed in lots of Trifolium repens. In Silene gallica, a reasonable prediction of contamination was found.

7 27th ISTA Congress – Seed Symposium

TESTING FOR ADVENTITIOUS PRESENCE OF TRANSGENIC MATERIAL IN CONVENTIONAL SEED LOTS USING QUANTITATIVE LABORATORY METHODS: A NEW STATISTICAL APPROACH AND ITS IMPLEMENTATION LAFFONT1,*, JEAN-LOUIS, REMUND2, KIRK M., WRIGHT3, DEANNE L. & SIMPSON2, ROBERT, D.

1Pioneer Génétique, Chemin de l’Enseigure, 31840 Aussonne, France 2Monsanto Company, 800 North Lindbergh Blvd., St. Louis, Missouri, 63167, USA 3Pioneer Hi-Bred International Inc., 7300 N.W. 62nd Avenue, Johnston, Iowa, 50131-1004 USA *[email protected]

The statistical methodologies used in testing for Adventitious Presence of transgenic material in conventional seed lots are well defined when the laboratory methods employed are qualitative (the response is the presence or absence of a particular trait). These methodologies are implemented in numerous applications, among which we can mention the freely available Microsoft Excel® spreadsheet application Seedcalc. When the response from the method used by the laboratory is quantitative (e.g. percent DNA of the transgenic material), the statistical methodologies developed for qualitative laboratory methods are not fully appropriate; although we can use the generally well known concepts from the area in Statistics called Statistical Quality Control in both cases, the nature of the errors and the underlying distributions involved are not the same for the two types of laboratory methods.In this paper, we present the details of a new approach specific to quantitative laboratory methods. In particular we will consider: - the assessment of quantitative laboratory method errors using linear modeling; - the decision making process, including details of the probability calculations needed to develop Operating Characteristic Curves and estimate consumer and producer risks for a given Lower Quality Limit (LQL) and Acceptable Quality Limit (AQL) depending on the testing plan (number of pools and number of seeds per pool, laboratory errors, acceptance threshold). We will also describe their implementation in the newest version of Seedcalc, Seedcalc6 which has capabilities for assessing quantitative laboratory errors and for finding optimal testing plans.

BIOCHEMICAL CHARACTERIZATION OF WHITE ONION ECOTYPES (ALLIUM CEPA L.) THROUGH HPLC ANALYSIS OF SEED STORAGE PROTEINS MENNELLA, G., ONOFARO SANAJA’, V., D’ALESSANDRO, A. & MILONE, M.

Istituto Sperimentale per l’Orticoltura – MiPAF, Via Cavalleggeri 25, 84098 Pontecagnano (SA), Italy, [email protected]; [email protected]

Water-, salt-, alcohol– and alkali- soluble seed storage proteins, extracted from 21 “Cipolla bianca di Pompei” onion ecotypes (Allium cepa L.), were analysed by anionic exchange-high performance liquid chromatography (AE-HPLC), sodium dodecil sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and isoelectrofocusing (IEF). AE-HPLC elution profiles (time range 0 to 40 mins) at 280 nm of water–soluble proteins evidenced the presence of 21 peaks, which allowed all the ecotypes studied to be distinguished from each other. The differences detected were both qualitative (presence/absence of one or more peaks) and quantitative; the water–soluble proteins were useful in differentiating ecotypes and cultivars while the other seed protein fractions only showed a weak polymorphism. The dendrogram analysis, based on HPLC data, showed that the ecotypes clustered with a genetic similarity degree of about 69%. The possibility of discriminating between closely related onion ecotypes during the course of breeding programmes could allow the identification of biochemical markers linked to useful agronomical traits. SDS–PAGE and IEF showed a minor degree of resolution and did not allow to identify unequivocally all the ecotypes studied. As observed by chromatographic analysis, the globulin composition of onion water–soluble seed protein appears to be independent of environmental growth conditions.

8 Budapest, Hungary, May 17th – 19th, 2004

The biochemical characterization of the available typical onion germplasm may contribute to obtain a community recognition and denomination, such as Denomination of Protected Origin (D.O.P.), Indication of Protected Origin (I.G.P.), Specificity Attestation (A.S.). The biochemical method here developed resulted of high resolution, cost-effective and time- saving for characterization and genetic purity assessment of the ecotypes studied.

COMMERCIAL SUNFLOWER GERMPLASM IDENTIFICATION AND CHARACTERIZATION USING SSR. VICARIO, A.1, LORAY, A.1, PANIEGO, N.2 & HOPP, E.2

1Laboratorio de Marcadores Moleculares. Dirección de Calidad. Instituto Nacional de Semillas. Secretaría de Agricultura, Ganadería, Pesca y Alimentos. Av. Paseo Colón 922, 4to piso. Ciudad de Buenos Aires. Argentina. 2Instituto de Biotecnología CICVyA-CNIA-INTA Argentina. CC 25, 1712 Castelar, Argentina

World oilseed production has risen up to 327.80 million ton in the last few years. For sunflower this world annual production is 24.02 million tons. Argentina is the fourth oilseed producer and the first sunflower producer in the world. Sunflower is the second oilseed produced in Argentine and the forth commodity produced after soybean, corn and wheat with an expected production of 4.2 million ton on 2.3 million of hectares for the 2003/2004 campaign. In spite of its importance, molecular characterization and identification of commercial germplasm using modern DNA marker technology is still scarce. In this work we selected a group of 10 hypervariable SSRs to assess their ability for identification, characterization and quantification of the genetic diversity on 27 representative sunflower hybrids from Argentina. PCR amplification products were resolved by standard sequencing electrophoresis on denaturing polyacrylamide gels and revealed using a silver nitrate staining kit. We observed a mean of 5.7 alleles per locus and the presence of at least one heterocygote locus in all hybrids analyzed. The total diversity was 0.54. All genotypes could be differentiated using 10 SSR. More studies applying these new markers will be carried out.

DEVELOPMENT OF MICROSATELLITE MARKERS FOR THE IDENTIFICATION OF BRAZILIAN COFFEA ARABICA CULTIVARS. VIEIRA, ELISA S.N.1, VON PINHO, ÉDILA V.R.1, ESSELINK, DANNY G.2, VIEIRA, MARIA G.G.C.1 & VOSMAN, BEN2

1Federal University of Lavras, Lavras, Minas Gerais, Brazil, [email protected] 2Plant Research International, Wageningen, The Netherlands

Microsatellite markers or SSR (Simple Sequence Repeats) have proved to be an excellent tool for cultivar identification and genetic relationship studies in plants. A set of 140 SSR markers were used to analyze the genetic similarity among twenty five Coffea arabica cultivars composed of nineteen brazilian cultivars of commercial importance, and six indian hybrids of Coffea arabica, Coffea canephora and Coffea liberica. Of the total number of SSR markers tested 127 DNA nuclear markers were developed using enriched small insert libraries and the coffee SSR sequences available in the NCBI database, and thirteen universal chloroplast DNA markers were also tested. Among the twenty-two polimorphic loci 2-7 alleles were detected for each locus with an average of 3.5 alleles per locus. The locus CAga001 was the most discriminating for brazilian cultivars with 4 allelic phenotypes and 1.9 effective alleles. The mayority of the SSR loci contained di-nucleoditde (GT) repeats and the polymorphism was positively correlated with the number of repeats. Based on the band patterns generated by the polymorphic SSR loci, the set of twenty-five coffee cultivars were clustered in two main groups. One group composed of the most part of the brazilian cultivars and a second one composed of the indian hybrids. Many of the color mutants were not separated. The clustering was in accordance with the background of the cultivars and showed the high level of similarity of the brazilian material, indicating the necessity of new introductions or crosses between cultivars with different background in future breeding programms.

9 27th ISTA Congress – Seed Symposium

SESSION 2 – Organic and Conventional Seed Production Chaired by Jose França Neto, Brazil

VARIETAL CHARACTERIZATION AND GENETIC PURITY ASSESSMENT OF CASTOR (RICINUS COMMUNIS L.) GENOTYPES ANKAIAH, R., MANOHAR REDDY, N., KESHAVULU, K., SAMBASIVA RAO, P. & MURALIMOHAN REDDY, B.

National Seed Project, ANG Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, India, [email protected]

The investigation on varietal and genetic purity assessment of castor genotypes was conducted to develop field and laboratory techniques to distinguish genotypes. The experimental material comprised of nine castor hybrids (GAUCH-1, GCH-2, GCH-4, PCH-1, TMVCH-1, DCH-32, GCH-5, DCH-177and GCH-6), their parents (VP-1, VI-9, JI-35,48-1, PCS- 136, LRES-17, TMV-5, DCS-5, GEETHA, SH-72, DPC-9, DCS-9, JP-65 and JI-96) and four varieties (Aruna,AKC-1,GC-2 and Kranti) for study .during 2001 and 2002. Various chemical tests (biochemical techniques electrophoretic technique of total soluble seed proteins) were used to characterize the hybrids and their parents at laboratory level. Distinct, uniform and stable morphological characters were identified for nine hybrids, their parents and four varieties with the help of 38 morphological characters which were least influenced by growing conditions. Based on morphological characters, flow chart which is useful for field functionaries involved in seed production/certification for easy identification of castor genotypes was developed. Grouping of castor genotypes was possible based on differential growth response of seedlings to GA3 and seedling sensitivity to 2,4-D. The electrophoretic banding pattern of each genotype was unique and distinct between hybrids, their parents, and varieties for total soluble seed proteins. These differences were useful in identification of individual castor genotypes. The genetic purity tests by field plot technique (GOT) based on distinctness, uniformity and stability of morphological characters was found to be a foolproof method for estimating genetic purity.

ORGANIC SEED TREATMENT TO CONTROL COMMON BUNT (TILLETIA TRITICI) IN WHEAT BORGEN, ANDERS

Houvej 51, 9550 Mariager, Denmark

Common bunt caused by the fungus Tilletia tritici (syn. T.caries) is one of the most devastating plant diseases in wheat. In conventional agriculture the disease is controlled exclusively by fungicide seed treatment, but in organic farming these fungicides are not accepted. Previous studies in India have shown that seed treatment with plant extracts of Canabis sativa, Eucalyptus globulus, Thuja sinensis and Datura stramonium was fully effective against the disease under field conditions. Later, in vitro studies have shown that also germination of spores of the Karnal bunt pathogen (Neovossia indica) could be prevented by these plant extracts. The experiment was repeated in Denmark with extracts from the same species grown in Denmark, which has climate conditions very different from India. In this experiment the same seed treatments had no or very limited effect on the frequency of the disease. The treatments were compared with indigenous methods from Europe including salty brine, Thuja leaves and lime. These methods had a significant but insufficient effect on disease suppression.

10 Budapest, Hungary, May 17th – 19th, 2004

CHALLENGES FOR OBTAINING HIGH QUALITY ORGANIC SEEDS GROOT, STEVEN P.C. & VAN DER BURG, W. JOOST

Wageningen Seed Centre, Wageningen University and Research Centre, P.O. Box 16, NL 6700AA Wageningen, The Netherlands, www.seedcentre.nl

High quality seed is the basis of crop production. Seed companies perform checks and treatments during seed production, and after harvest, to guarantee they provide their customers with good, germinating, healthy seeds. Many countries have established seed testing stations for certification and control of seed moving in trade. Quality of seeds has many aspects, these include authenticity of the variety, purity of the seed batch, its germination potential and seed health. For organic farming additional aspects of seed quality are important, especially the production of seeds under organic farming conditions and the restriction in methods that can be used for the treatment of the seeds. Rules for organic crop production are made by international bodies such as the International Federation of Organic Movements (IFOAM) and the EEC (Regulation 2092/91). Certification for ecological production is performed by appointed national organisations, distinct from organisations that certify seeds on the quality aspects mentioned above. Production and treatment of seeds under organic conditions puts additional challenges for obtaining high quality seeds. Because of the limitations in the use of chemicals, organic production bears a greater the risk of contamination with weed seeds or seed contaminated with pathogens. Moreover, sowing of seeds in soils with organic fertilisers that have slower mineralisation in spring, and weed competition may ask for seedlings with a faster growing root system: seeds with additional quality traits such as seed vigour. Presently it is debated whether organic seeds would need lower thresholds for contamination with GMO seeds than non-organic seeds. These challenges urge for finding solutions, for a large part through research. Adequate methods for the detection of pathogens during seed production and analysis of critical control points will provide the basis for disease monitoring activities and treatments. Alternative seed sanitation treatments need to be developed as alternatives for the present use of fungicides. These treatments need not only be effective in elimination of the pathogen, but should also maintain the viability of the seeds and be acceptable for both the standards for organic farming and (inter)national regulations regarding the use of components for crop protection. Novel seed sorting techniques may be of use in sorting out diseased seeds. For those crops where at present no economically feasible alternatives are available the effect of increasing the disease threshold for organic seeds is under discussion. Examples will be provided of seed research in response to these challenges for obtaining high quality organic seeds.

A COMPARATIVE STUDY OF THE GERMINATION CHARACTERISTICS OF WILD FLOWER SEEDS IN COMMERCIAL SEED PRODUCTION KHAJEH-HOSSEINI1, MOHAMMAD, POWELL1, ALISON A. & LAVERACK2, GILES K.

1Department of Agriculture & Forestry, University of Aberdeen, Hilton Campus, Hilton Place, Aberdeen, AB24 4FA, Scotland, U.K 2Scotia Seeds, Mavisbank, Farnell, Brechin, Angus, DD9 6TR, Scotland, U.K

The demand for, and production of, wild flower seeds has increased in recent years, but there has been little attempt to test germination and breaking dormancy of cultivated seeds of these species. Therefore, the germination of seeds from sixty species of cultivated wild flowers produced in Scotland was examined for two years of seed production (2001 and 2002). The seed moisture content (MC) and laboratory germination were determined and four methods of breaking dormancy: prechilling, scarification, scarification with high temperature and gibberellic acid (GA3) were used depending on the species. The seed MCs ranged from 4 to 12% and were higher in most species in 2002 when seed had been stored at a higher relative humidity (45% compared with 35% in 2001). The total germination and percentage normal seedlings of species ranged between 0-100% (2001) and 0-91% (2002)

11 27th ISTA Congress – Seed Symposium although overall the number of normal seedlings was higher in 2002. Scarification increased germination in some Fabaceae species, whilst GA3 increased germination in other species. Seed surface sterilisation, to eliminate pathogens from the seed surface, and prehydration treatments, to eliminate imbibition damage during germination, also improved the germination in some cases. Individual species differed in their germination and dormancy in the two years of production. These differences are discussed in relation to differing weather conditions, methods and timing of harvesting, methods of seed cleaning and seed storage conditions.

THE EFFECT OF GLYPHOSATE TREATMENT ON THE GERMINATION POTENTIAL OF BARLEY SEED MCLAREN, GILLIAN & DON, RONALD

Official Seed Testing Station for Scotland, Scottish Agricultural Science Agency, East Craigs, Craigs Road, Edinburgh, EH12 8NJ, United Kingdom, [email protected]

Glyphosate applied to crops at ear moisture content greater than 35% had a detrimental effect on seed viability. Below 35%, germination in paper towelling was lower than in compost due to a higher number of glyphosate-induced abnormal seedlings. The number of glyphosate-induced abnormalities was not related to ear moisture content, but to the level of rainfall experienced in the period immediately after spraying.

EFFECT OF DESICCATION ON SOME QUALITY CHARACTERISTICS OF SUGARBEET SEED RAJIC, MILORAD, MARINKOVIĆ, BRANKO, MIKLIČ, VLADA & PANKOVIĆ, LAZAR

Institute of Field and Vegetable Crops, Novi Sad, Serbia and Montenegro, Faculty of Agriculture, Novi Sad, Serbia and Montenegro

A three-year field trial has been established in a system of random blocks in order to study the effect of desiccation on the 1000-seed mass and germinability of sugarbeet seed under the local climatic conditions. Desiccations were performed at the beginning and end of wax maturity. As the timing of cutting and threshing operations is important in the harvest of seed sugarbeet, desiccation simplifies the overall harvest. The application of desiccation under the local agroecological conditions has caused significant changes in the studied quality characteristics. The 1000-seed mass was significantly larger in the control than in the desiccation variants. The later application resulted in higher seed mass compared with the earlier application. Significant differences were also registered in seed germinability. It was therefore clear that the interruption of growing season negatively affected the major quality characteristics of sugarbeet seed. In view of the significant differences among the study years, the obtained results showed that seed sugarbeet responds uniformly to different climatic conditions.

12 Budapest, Hungary, May 17th – 19th, 2004

Keynote – Development of Education and Training in Seed Science and Technology By Murray Hill, New Zealand

DEVELOPMENT OF EDUCATION AND TRAINING IN SEED SCIENCE AND TECHNOLOGY HILL1, MURRAY J. & COOLBEAR2, PETER

1Professor of Seed and Crop Science and Director, New Zealand Seed Technology Institute, P O Box 84, Lincoln University, Canterbury, New Zealand, [email protected] 2Deputy Chief Executive, Manukau Institute of Technology, Private Bag 94006, Auckland, New Zealand, [email protected]

In this paper, we provide an overview of the factors that need to be considered in developing cost-effective, successful seed technology education and training programmes. While the benefits to the individual of quality programmes are self-evident, the key to maximising the impact of such education and training is in matching relevant training options with carefully selected trainees who are subsequently supported to maintain a hands-on leadership role within the industry. Our own survey data and subsequent anecdotal evidence emphasises the fact that providing the necessary managerial and infrastructural support is critical to the sustained application and transfer of new skills and knowledge. The international education and training effort in seed science and technology has lost much of its impetus since the early 1990s when aid funding declined in this area. We argue that it is timely for ISTA to utilise its combined expertise and take a much more proactive role in lobbying for resources to support effective international programmes.

SESSION 3 – Viability and Vigour: Evaluation and Impact Chaired by Joël Léchappé

VIGOUR TESTS FOR PREDICTING SEEDLING EMERGENCE OF AUBERGINE (SOLANUM MELONGENA L.) SEED LOTS DEMIR1, I., ERMIS1, S., OKÇU1, G. & MATTHEWS2, S.

1Department of Horticulture, Faculty of Agriculture, University of Ankara, 06110 Diskapi/Ankara, Turkey, Fax: 09 90 312 3170550/1316 2Department of Agriculture, University of Aberdeen, Aberdeen, Scotland, United Kingdom

The seed vigour of 5 aubergine (Solanum melongena L.) seed lots was assessed by cold, conductivity and controlled deterioration tests and compared with seedling emergence in an unheated glasshouse. Germination of aubergine seeds after controlled deterioration (CD) (range 6 to 69 %) for 24, 48 (2 to 83 %) and 72 hours (2 to 81 %) at 45°C at 20 % moisture were positively and significantly related to seedling emergence, the lots being ranked consistently the same for all durations of CD and glasshouse emergence (42 to 93 %). CD germination was more closely related to glasshouse emergence after 24 (R2 =0.972, P<0.01), 48 (R2 =0.970, P<0.01) and 72 hours (R2 =0.0.968, P<0.01) compared to the standard germination (range 72 to 94 %), (R2 =0.921, P<0.01). The cold and conductivity results were not significantly related emergence. The CD test is suggested as method to predict emergence in aubergine which should be developed further.

13 27th ISTA Congress – Seed Symposium

RELATIONSHIP BETWEEN STANDARD AND COLD GERMINATION TESTS IN SUPERSWEET SWEETCORN GEORGE, D.L., GUPTA M.L. & PARWATA, I.G.M.A.

School of Agronomy and Horticulture, University of Queensland, Gatton, Qld, 4343 Australia, [email protected]

Seed viability of sweetcorn is usually measured by the standard germination test with the germinator set at 20 - 30oC. The cold test, so named because it is performed at 10oC, is used to assess seed vigour through temperature stress. It has been found most suitable for seed with high standard germination (>85%). Both tests were performed on 504 samples of seed of the supersweet sweetcorn cultivar, Pacific H5 of which 162 samples had a standard germination higher than 85%. The crop was planted in spring 1998 in the Lockyer Valley, Queensland and cobs harvested at two seed moisture ranges (20-30% and 40-50%) were dried under different combinations of drying temperatures (30°C, 40°C and 50°C) and air velocities (1.25 m/s, 2.75 m/s and 4.30 m/s). Cobs were then shelled and cleaned manually after which the seed was stored at 30oC. Germination tests were conducted at two monthly intervals from June 1999 to April 2000. The mean and range for cold germination (Gc) was 81% and 72 - 90% while for standard germination (Gs) it was 90% and 85 - 98% for 162 samples. The relationship between Gc and Gs (>85%) was best explained by the equation Gc 2 = -21.0 + 1.14 Gs (R = 0.62).

ASSESSMENT OF VIABILITY OF BREAD WHEAT (TRITICUM AESTIVUM L.) AND OATS (AVENA SATIVA L.) GERMPLASM SAMPLES SORED OVER 30 YEARS IN COLD STORE HOLLY, LÁSZLÓ, BÓCSÓ, RENÁTA, JUHÁSZ, ATTILA & MÁR, ISTVÁN

Institute for Agrobotany, Tápiószele, Hungary, [email protected]

Medium- and longterm storage of seed samples is a key element of the conservation of crop genetic resources collections. Preservation of seeds in cold stores at low moisture content/relative air humidity can extend the storage life of samples and reduce the frequency of regenerations required. It contributes, therefore, to the maintenance of the original genetic structure of populations in PGRFA collections. The first two cold storage rooms for genetic resources conservation at the Institute for Agrobotany were established in 1973. At the beginning, mainly cereal (wheat, barley and oats) seed samples were placed into the cold stores. In accordance with the international standards for germplasm conservation, seed samples were dried at room temperature under low (15-30 %) relative air humidity and airtight containers (glass jars) were used for medium-term storage. Viability of seed samples was assessed before storage and monitored at regular intervals by standard germination tests. Seed samples have been regenerated whenever their viability dropped below 70%, or the quantity of seed decreased to a minimum leve1. The original seed samples were, however, not discarded hut kept for further studies until the complete lass of their viability. After 30 years of cold storage, 141 wheat and 132 oats samples still survived and 30 of them were selected for further studies. The storage behaviours of the two species show same distinct differences. The variation among seed samples in storability was more pronounced in wheat, while the oats samples perform more uniformly during storage. The analysis of the influence of the initial gen11ination percentage in wheat on the longevity in cold store revealed strong correlation between the initial germination and the values obtained in the tests conducted after 14 and 28 years of storage, r= 0.5506 (n=141) and r= 0.468 (n= 141), respectively. The same correlation coefficients for oats were r= 0.208 and r= 0.106 (n=132). Some seed samples of wheat (18) and oats (32) had an initial germination percentage lower than 85 %. In these sample groups, correlation was not found between the initial and the last germination test results (r= 0.067 and r= 0.071). It is assumed that in these cases the initial viability was underestimated probably due to incomplete after ripening and unresolved partial dormancy of seeds. This assumption is supported by the slight increase of germination percentage observed in the second control germination test in certain 14 Budapest, Hungary, May 17th – 19th, 2004 accessions. It is concluded that under the storage conditions applied (<8% seed moisture content, 0 C° storage temperature), the first control germination test should only be scheduled for after 15 years of storage, but the subsequent tests must be done in five years intervals in the two species studied. Considerable amount of seed could be saved by introducing this viability monitoring system. The increase in standard deviation of germination percentages for both species indicates that the genotype and perhaps genotype x harvest year interaction influence the longevity of stored seeds. A set of 15 seed samples was selected from both species for further studies in order to test germination speed and vigour and field emergence after 30 years of' storage. An analysis of changes in the genetic structure of stored samples is also underway.

ASSESSMENT OF RELIABILITY OF GERMINATION PAPERS IN ENVIRONMENTAL STRESS RESEARCH KHAJEH-HOSSEINI1, MOHAMMAD, POWELL1, ALISON A. & BINGHAM2, IAN

1Department of Agriculture & Forestry, University of Aberdeen, Hilton Campus, Hilton Place, Aberdeen, AB24 4FA, Scotland, United Kingdom 2SAC, Department of Agronomy, Craibstone Estate, Aberdeen, AB21 9YA, Scotland, United Kingdom

The substrates used for seed germination in experiments investigating the effect of stresses, such as salinity, on germination and early seedling growth, are usually assumed to have no effect on the results of the experiments. Our concern is that in some cases the germination substrate, such as paper towels, may influence the results. The effect of salinity on the early seedling growth of soybean (cv. Williams) was therefore studied using a range of NaCl concentrations (0-161 mM NaCl) using two methods, paper towels and a hydroponic system. Seeds were germinated in paper towel at different salinities (0-161 mM NaCl) then transferred to either a new paper towel or the hydroponic system at the same salinity. Seedlings were harvested 10d after transfer and both fresh and dry weight assessed. The concentrations of Na+, K+ and Ca2+ in the seedling and in the solution extracted from paper towels were determined by Flame Photometry and Atomic Absorption Spectrophotometry. Seedling growth was reduced with increased salinity, but the effects were markedly less when experiments were conducted in paper towels. Seedlings were able to grow in paper towel at an external NaCl concentration of 161mM NaCl, when a Na+ concentration 2.3mg g-1 FW was recorded in the roots. However, in the hydroponic system, seedlings failed to grow even at an external NaCl concentration as low as 12 mM, when the Na+ concentration in the root was 1.6mg g-1 FW. Analysis of the solutions soaking the paper towels revealed that 4.25mM Ca2+ was available to the seeds in the paper towel system in saline conditions. Enhanced growth was observed in the hydroponic system at 30mM NaCl following addition of 4.25 mM Ca2+. This suggests that paper towels may influence the response of seedlings to NaCl due to the release of other ions from the paper towels into the solution soaking the towels. The role of Ca2+ in reducing Na+ uptake or protecting the seedling against the toxic effects of Na+ is discussed.

PERFORMANCE OF OSMOPRIMED SEED OF GERMPLASM OF DESI AND KABULI CHICKPEAS UNDER LABORATORY AND FIELD CONDITIONS SINGH, S.J.& SINGH, KARAN

Rajasthan Agricultural University, Agriculture Research Station, Durgapura-Jaipur-302018, India

Investigation on laboratory performance of osmoprimed seeds of K-850, GNG-469, Phule G- 12 and RSG-143.1 genotypes of desi and ICCV-2, L-55, KAK-2 and Pusa-1003 genotypes of Kabuli chickpeas (Cicer arietinum L.) were conducted. Seeds were subjected to osmoprimal invigoration treatment by using polyethlene glycol ‘6000’ osmotica of different osmotic potential for various duration at 25 ± 1°C befxore storage and redrying. These seeds were evaluated for germination percentage, germination relative index, seedling length, seedling vigour index, seedling emergence index and electrical conductivity of seed leachates (as an index of seed vigour). It was observed that Osmo primal invigoration treatment (with PEG 15 27th ISTA Congress – Seed Symposium

6000) given to seeds for 18 hours (germplasm of desi chickpea) and 12 hours (germ plasm of kabuli chickpea) at 25 ± 1°C was effective in improving the storability and laboratory performance of seeds. However osmotic potential (negatively) higher than 7.5 bars (desi genotypes) and –5.0 bars (kabuli gram genotypes) caused deleterious effects on various seed technological parameters an initial testing (7 days after treatment and redrying) and on testing after 18 months of insect free ambinent storage. Treatments given to seeds for longer hours (exceeding 18 hours in desi and 12 hours in kabuli) adversely affected the seed and seedling vigour in chickpeas. Chickpea genotypes belonging to desi (Microsperma) and kabuli (Macrosperma) showed variability with respect to their responses to the treatment. Use of commercial grades of carbowox (PEG 6000) for osmoprimal in vigoration of seeds of chickpea has been advocated for improving the seed quality and storability as evaluated under laboratory conditions and also under field conditions (reported in a separate research paper).

VIGOUR TESTING: TOWARDS AN EXTENDED USE OF THE CONDUCTIVITY TEST WAGNER, M. HELENE, PREVEAUX, ANNE, MOIZAN, ELISE, BEAULATON, MATTHIEU & DUCOURNAU, SYLVIE

GEVES-SNES, rue Georges Morel, B.P. 24, 49 071 Beaucouzé Cedex, France

Bulk conductivity test on garden pea was introduced in ISTA Rules in January 2002. To get a larger useful test, we try to extend it to other species among the most cultivated in the French production: wheat, maize, rapeseed, sunflower, and lucerne. Five species were chosen because of their different storage tissues: oil crops, starchy ones and legume and because they represent different kind of botanical seeds: akene, caryopsis and single seed. Five samples were used by specie and analysed with a conductivity meter fitted with a dip cell having a cell constant 1 cm-1. The temperature of the conductivity test was fixed at 20°C like for pea but the other parameters of the test were adjusted to the size of samples and to the kind of seed storage tissues. Finally, three methodologies can be suggested for rapeseed, wheat and maize, which are accurate and repeatable and which can classify seed lots more tightly than germination test. They should now be validated with comparative tests between different laboratories and with field emergence results. Some work is still to be done on sunflower because of the electrolytes leakage from the pericarp more or less important belong to the variety. For lucerne seeds, so much variability is met with hardseededness and to solve this problem, recovery and weighing of hard seeds is not enough.

SESSION 4 – Seed Systems in Emerging and Development Economics Chaired by Grethe Tarp

SEED SECTOR DEVELOPMENT AND SEED POLICIES IMPLEMENTED IN TURKEY BOZKURT BAHATTIN , YALVAC KENAN* & BAL O. FARUK

*Tarim Ve Köyişleri Bakanliği, Milli Müdafaa Caddesi No 20 Kat 8 O, Kizilay, Ankara, Turkey [email protected]

The improvement and development of the seed sector is affected by the policies identified and implemented by the Ministry of Agriculture in Turkey. Recent developments in science and technology have been largely put into practice in two main sectors of agriculture; plant breeding and seed sectors. The seed policy of the Ministry aims at developing a modern seed industry led by the private sector with its institutions allowing technology transfer and putting more emphasis on the seed production in Turkey.

16 Budapest, Hungary, May 17th – 19th, 2004

SEED SECTOR DEVELOPMENT IN SOUTH WESTERN NIGERIA: FARMERS’ UTILIZATION OF IMPROVED SEED FOR CROP PRODUCTION DANIEL, I.O.1 & ADETUMBI, J.A.

1Department of Plant Breeding & Seed Technology, University of Agriculture, PMB 2240, Abeokuta, Nigeria

Farmers’ acquisition and utilization of seeds of improved varieties for crop production in South Western Nigeria was investigated to characterise the local seed system and assess the potentials of the seed sector. Using structured questionnaires, a seed survey was conducted involving 94 farmers randomly sampled from 5 farm settlement locations covering 3 states in the region. The data collected were subjected to descriptive analysis of simple proportions and percentages. Four of the most propagated crops of the region were maize, cassava, cowpea, and different kinds of vegetables. For all the predominant crops that were cultivated at the survey sites, percentage of total land area cultivated with the seeds of improved varieties was over 90% throughout the 3 years surveyed. Over 50% of farmers saved seeds of improved varieties for subsequent cropping from previous harvests, indicating high adoption rates but low replacement rates. Moreover, most farmers would purchase seeds of improved varieties if it would lead to increased yields and meet consumer preferences while a negligible number of farmers would purchase seed because of low costs, meaning that farmers are ready to pay for seeds of improved varieties, if the seeds are true to their labels. The implications of the results on seed industry development and suggestions for strengthening of the commercial seed sector are discussed.

SEED PROGRAMME DEVELOPMENT IN A TRANSITION ECONOMY – THE EXPERIENCE OF VIETNAM THANH DING NHAT DUNG & TURNER, MICHAEL

Danida-ASPS, Seed Component, A6, Ministry of Agriculture and Rural Development, 2 Ngoc Ha St., Ha Noi, Viet Nam, [email protected]

For the past 30 years seed projects have been implemented in many ‘developing countries’ with external support from donors and with the aim of establishing the basic elements of a formal seed supply system. Within the past decade, there have been similar initiatives in the ‘transition economies’, mostly in the countries of Eastern Europe and the Former Soviet Union. Here, the context is very different because a seed system already existed but it was part of a planned economy and was driven by production targets, not by market demand. Vietnam is also in transition to a market economy, but its agricultural sector is more typical of a developing country and, like much of South-east Asia, it is now experiencing rapid economic growth. It therefore presents an interesting case study in terms of seed sector development, for which support is being provided by Danida This paper explains the technical and economic factors which influence agricultural development in Vietnam, resulting mostly the predominance of rice as the staple crop. Against this background, some key issues are discussed in relation to the changing role of the State from a major seed producer, to being the regulator for a more diverse seed sector. The transition process has particular implications for quality control/assurance as new seed suppliers seek to establish a reputation for quality and need access to independent seed testing facilities. Finally, the paper notes some of the policy issues which arise during the transition process and the need to have a mechanism for resolving those issues in an equitable and consistent way.

17 27th ISTA Congress – Seed Symposium

APPLIED TREE SEED TECHNOLOGY IN BRAZILIAN ATLANTIC FOREST FATIMA C.M. PIÑA-RODRIGUES & E.S. NOGUEIRA

Universidade Federal Rural do Rio de Janeiro- BR 465 km 7- Seropedica- RJ- Brazil- CEP: 23851970, [email protected]

During the last 15 years brazilian research on seed science technology concentrated in exotic species as Eucalyptus and Pinus. The Brazilian Rules for Seed Analysis present recommendations for many exotic species, with the insertion of few brazilian ones. In order to analyse the status of tropical seed research in Brazil, the objective of this work was to carry out a survey of the state of art of the research, production and technology of forest seeds. Survey was developed using bibliographical consultations, web tools searches; information and documentations rescue and technical visits and questionnaires. A great number of exotic species are included in the most produced. Although there is an equilibrium between the number of species researched (n= 331) and produced (n= 364), only 7.6% (n= 53) of cited species are being studied and produced at the same time. The most produced species are used for restauration of degraded areas and homogeneous plantations (38%). Arborization and ornamental objectives were dominant in relation to the others uses (62%). Maturation, pathology, phenology and harvest presented the lowest number of researches references (< 1.3%) while storage was the most searched. From 212 published scientifical works, only 19.6% studied methodologies for forest seed analysis, involving less than 10% of the more produced species. Only 23.9% of published works approach the subject dormancy. Methodologies to standardization of germination assays are necessary to inclusion of the species in the Brazilian Rules for Seed Analysis because of the lack of researches with this specific finality.

HIGH QUALITY RICE SEED PRODUCTION - SANTA CATARINA MODEL KNOBLAUCH, RONALDIR

Agricultural Research and Extension Service Agency for Santa Catarina State: EPAGRI/Itajaí Experiment Station, Rodovia Antônio Heill, km 06 - CEP: 88301-970 - ITAJAÍ, SC. Brazil, [email protected]

Rice production in Brazil averages 10.5 million tons/year. The state of Santa Catarina located in Southern Brazil has an area of 134 thousand hectares of irrigated rice. In 1976, rice productivity was 2.3 tons/ha; last season, 2002/03 the mean yield, reached 7.5 ton/ha, the highest in the country. This yield increase is attributed to several factors, but it is mainly due to the release of high yielding cultivars put out by Epagri, the Research and Extension Service for Santa Catarina and also due to implementation of a high quality seed program in 1986. This program has the objective to reduce the number of red rice in the classified seeds as a mean to control red rice in the fields. Until 1970 it was allowed 20 seeds of red rice per 500 grams sample. The step by step reduction in the number of red rice in the seed lots, came to a zero seed/500 gram sample for all the classes of seeds, started in the year 2000. In 1986 only 20% of the state rice farmers had grown rice from certified seeds; at the present this number came to over 80%. The state seed growers are now exporting seeds to different countries in South America and to most rice growing areas of Brazil.

18 Budapest, Hungary, May 17th – 19th, 2004

POTENTIAL OF ON-FARM PRACTICES FOR IMPROVING RICE SEED QUALITY, SEED HEALTH AND CROP PRODUCTION S.B. MATHUR1,S. B. C.N. MORTENSEN1, C. N., M.H. TALUKDER2, R.B. MABAGALA3

1Danish Government Institute of Seed Pathology for Developing Countries, Thorvaldsensvej 57, DK- 1871, Frederiksberg C, Denmark, [email protected] 2Department of Agriculture Extension, Ministry of Agriculture, Government of Bangladesh, Khamarbari, Farmgate, Dhaka-1215, Bangladesh 3Department of Crop Science and Production, Sokoine University of Agriculture, P.O. Box 3005, Morogoro, Tanzania, [email protected]

Investigations conducted in Bangladesh and Tanzania showed that rice seed lots often contain shrivelled, spotted and discoloured seed. Seed with such physical deformities are often lighter and infected by several important fungal and bacterial pathogens. Removal of these types of affected seed has led to increased seed germination, production of healthier seedlings, resulting in higher yields. Removal of seed of poor quality was conducted by manual sorting or by the flotation technique using a 15% sodium chloride solution (brine solution). In separate trials (3-years of data) conducted in the field in Bangladesh the average germination of rice seed increased from 73% to 85% and resulted in a 12.8% yield increase. In Tanzania, the germination was higher in sunken seed (94%) than in floating seed (39.4%). A higher incidence of fungal pathogens was observed in seed that floated in 15% sodium chloride solution than in seed that sunk; the use of sunken seed for planting gave yield increases of up to 34.6% compared to the untreated control. Additionally, the seedling emergence in seedbeds under farmers conditions revealed that the germination of seed separated by the sodium chloride solution was of more than 90% while only 85% germination was recorded when the seed was sorted manually. The percentage of germination in floating seeds was 0.5% and 30% in shrivelled, spotted and discoloured seed. The germination of the untreated control was 80%. Approximately 20 hours were required for four persons to remove discoloured and spotted seed by hand from 50 kg, which is the amount of seed required for sowing one hectare in Bangladesh. Sorting of seed by the 15% salt technique took less time than manual sorting. The time taken by individual farmers to separate 500 g of seed by the two technologies was evaluated in Tanzania; farmers’ treatment of seed with the brine solution required 5 min, while the sorting of the seed by hand took approximately 45 minutes. These simple technologies appear to be promising for farmers to separate good seed from poor quality seed and to select rice seed lots for planting. These technologies have been introduced to farmers in Bangladesh and Tanzania.

SESSION 5 – Seed Lot Hygiene Chaired by Akos Mesterhazy

FACTORS AFFECTING THE OCCURRENCE OF FUSARIUM SPP. IN CEREAL SEEDS IN NORWAY BRODAL, GURO1 & ELEN, OLEIF2

1Norwegian Agricultural Inspection Service, Seed Testing Laboratory, Box 3, N-1431 Ås, Norway, [email protected] 2The Norwegian Crop Research Institute, Plant Protection Centre, Høgskolevn 7, N-1432 Ås, Norway

The occurrence of Fusarium (represented by Fusarium spp. and Microdochium nivale) has been recorded on barley, oats and wheat seed in Norway since the 1970-ies as part of the seed quality assessment. Annually a large number of samples representing both certified and farm saved seed from all cereal growing areas, was tested (100 or 200 seeds from each sample) at the Seed Testing Laboratory using the freezing blotter, the Doyer filter paper, and the agar plate (PDA) methods. The Fusarium frequencies were recorded in each sample, a mean of all samples was calculated for each year for each cereal species, and multiple

19 27th ISTA Congress – Seed Symposium regressions with weather data and also the Fusarium incidences the previous year, were carried out. 69% of the variation of the incidence of Fusarium in barley was explained by 1) precipitation in July, 2) the incidence of Fusarium the previous year (significant positive effects), and 3) precipitation in June, 4) temperature in July (significant negative effects). In oats precipitation in July and incidence of Fusarium the previous year explained 59% of the variation. In wheat, as for barley and oats, precipitation in July and the incidence of Fusarium the previous year contributed positively, whereas precipitation in May and temperature in August had a negative effect. These four factors contributed significantly by 58% to the variation of Fusarium in wheat. It is concluded that the incidence of Fusarium the previous year and precipitation in July were the main factors affecting the incidence of Fusarium in cereal seeds in Norway.

USING SEED HEALTH TESTING RESULTS TO MAKE BALANCED SEED MANAGEMENT DECISIONS COCKERELL, V. & MCNEIL, M.

Scottish Agricultural Science Agency, East Craigs, Edinburgh, EH12 8NJ, United Kingdom, [email protected]

To be able to interpret seed health testing results it is important to understand the accuracy and limitations of different test methods. The relationship between traditional and Real-time PCR seed health testing methods for Microdochium seedling blight and bunt are described. Real-time PCR methods tend to indicate higher infection levels than traditional and could increase the chance of results being reported above recommended thresholds. However, they can provide growers with quick information that can be used to manage seed treatment decisions where time is limited. Traditional methods are best used where growers are not restricted by time or require a more precise indication of infection/contamination levels.

MYCOLOGICAL ASPECTS OF ONION SEED PRIMING DORNA1, H., TYLKOWSKA1, K., MARCINEK1, R., SZOPIŃSKA1, D. & WEI YAHONG2

1Department of Horticultural Seed Science, Technology and Nursery, the August Cieszkowski Agricultural University, Poznań, Poland 2Northwest Sci-Tech University of Agriculture and Forestry, Yangling, China

The purpose of the research was to study the changes in germination and the incidence of fungi occurring on onion seeds after their priming. Two samples of onion ‘Wolska’ seeds, differing in their initial quality, were used in the study. They were primed in polyethylene glycol (PEG 8000) at osmotic potential -1.5 MPa for seven days in darkness at 15ºC. In order to control the growth of fungi priming was combined with treating seeds with fungicides: Penncozeb 80 WP (a.i. 80% mancozeb) and Apron 35 SD (a.i. 35% metalaxyl). In combined treatments, the fungicides were applied before, during or after priming. Germination and mycological tests were performed at 10 and 20ºC. The seed samples differed in their response to priming both in terms of germination and presence of fungi. The treatments did not affect percentage of germinating seeds in most cases. Germination capacity was lower than percentage of germinating seeds. Priming and combined treatments increased germination rate. Seeds of a better germinating sample were infested with some pathogenic fungi, mainly with Botrytis aclada. From among saprophytic fungi, Penicillium spp. were the most common. Seeds of a worse germinating sample were colonized almost only with Penicillium spp. Species of Penicillium were dominating after priming. In general, combined treatments reduced their incidence. The lowest percentage of seeds infested with B. aclada was observed when fungicides were applied before priming.

20 Budapest, Hungary, May 17th – 19th, 2004

HEALTH STATUS OF SWEET CORN SEEDS IN CHINA AND ITS IMPACT ON SEEDLING EMERGENCE GUANGWU, ZHAO, GUOZHEN, ZHANG & JIANHUA, WANG*

College of Agronomy and Biotechnology, China Agricultural University, Beijing 100094, China, [email protected])

The health status of 18 sweet corn hybrids, collected from five locations in China, was examined and factors influencing seed health of sweet corn and relationships between seed health and seedling emergence were studied. Seven fungal genera on seed surfaces and 17 inside seeds were isolated. The number of fungi on seed surfaces and percentage of internally infected seeds were much lower in “Xixingtian1”, “Lutian9”, “Shanxitian” and “Xinjiangtian” than in “Jingketian115” and “Green superman”. Research showed that significant and consistent differences in the number of fungi on seed surfaces, in percentage of internally infected seeds and in fungal taxa inside seeds existed among three types. Sugar corn seeds were least infected while super sweet corn seeds most. Significant and consistent differences either in the number of fungi on seed surfaces or in percentage of internally infected seeds existed among five locations. Samples from Northwest China were least infected while those from Central China most infected. Further research showed that Fusarium was the most frequently isolated fungus and had the most drastic impact on seedling emergence (r=-0.555). Percentage of internally infected seeds had a significant negative correlation with percentage of seedling emergence (r=-0.489).

DISEASE FREE SEED PRODUCTION OF WHEAT IN PUNJAB (INDIA): ACHIEVEMENTS AND CONSTRAINTS SHARMA1, R.C., SHARMA2, INDU &. SAMRA1, J.S

1Seed Technology Center, 2Dept of Plant Breeding, PAU, Ludhiana – 141 004, INDIA [email protected]

Punjab Agricultural University, Ludhiana produces and supply about 3,000 tonnes of wheat seed (nucleus/breeder/foundation/certified), annually, to various public and private seed production agencies including farmers. Loose smut (Ustilago segetum var.tritici), Karnal bunt (Tilletia indica) and Ear cockle nematode (Anguina tritici) are designated as objectionable under the Indian Seed Act and minimum tolerance limits have been fixed for foundation and certified seed. The present studies were conducted to produce disease free seed by integrating existing and generating new and alternative management strategies. Nearly complete control of loose smut and cockle nematode has been achieved at seed production farms and farmers fields through chemical and mechanical seed treatments and farmer’s participatory campaigns. Carboxiin, carbendazim and recently the triazole compounds have proved highly effective against loose smut. The nematode galls are easily separated by the seed processing machines or by floating on brine solution. However, Karnal bunt has been an elusive problem where complete control could not be achieved. Even disease free areas could not be identified when extensive surveys were conducted. The disease prediction models have not worked satisfactorily in some of the seasons due to overall weather changes (long spells of fog during December-January) recorded in the recent years thus influencing greatly the disease epidemiology probably due to pre-ponement of teliospore germination in the soil The studies on teliospore germination, sporidial viability and multiplication carried out under field conditions have validated the above assumption. Nevertheless, a single spray of propiconazole/tebuconazole/hexaconazole at heading stage provided a considerable control in seed plots. In addition, resistant stocks to karnal bunt have been identified and are being used for incorporating resistance in high yielding wheats.

21 27th ISTA Congress – Seed Symposium

BLACK ROT ERADICATION TREATMENTS ON BRASSICA: EFFICACY AND SEED QUALITY INVESTIGATIONS TAYLOR1, A.G., KLEIN1,2 J.D., & MORRISON3 R.H.

1Department of Horticultural Sciences, NYSAES, Cornell University, Geneva, NY, USA 2Current address: Dept. of Field Crops, Volcani Center, Bet Dagan, Israel 3Sakata Seed America, Salinas, CA, USA

Black rot is caused by the pathogen Xanthomonas campestris var. campestris (Xcc) and is a devastating seed-borne disease on cabbage and other Brassica species. The objective of this research was to examine the efficacy of selected physical and chemical methods to eradicate the pathogen from an infested seed lots. Moreover, the effect of the eradication treatments were examined on the germination and seed quality. A traditional method to eradicate Xcc from Brassica seed lots is to employ a hot water treatment (HWT). Seeds are immersed in hot water (50°C) for a period of time ranging from 20 to 30 minutes to eradicate the pathogen. The effect of hot water (50°C for 20 minutes) was tested on a high and medium quality cabbage (B. oleracea var. capitata) seed lot. The germination percentage was not significantly reduced by the HWT in the high quality seed lot, while the HWT of the low quality lot resulted in a four percentage point reduction in germination. Further studies revealed that the HWT aged seeds to a similar extent as aging seeds at 70% RH (8.6% moisture content, fr. weight basis) and 40°C for a period of 7 days. Studies were conducted on selected seed treatments to eradicate Xcc from infested seed lots. In year one, a seed lot with external Xcc infestation and a seed lot with both external and internal infection received the following treatments: HWT treatments for 25 or 30 minutes (Reed’s Seeds, Cortland, NY, USA), a 1.0% Oxidate (hydrogen peroxide + peroxyacetic acid, BioSafe Systems, Glastonbury, CT, USA) soak for 4 hours at 25°C, and two proprietary methods – eXccit (Incotec, Salinas, CA, USA) and Pre-Prep (Seed Dynamics, Salinas, CA, USA). Pathological testing was performed in the third author’s laboratory, and testing procedures separated internal from external infection. All treatments eradicated Xcc from the seed lot with external infection, except for the Oxidate soak. The seed lot with both external and internal infestation was more problematic to eradicate. Both Oxidate soak and HWT for 25 minutes were not able to eradicate the external infection; however, the HWT for 30 minutes and all other treatments were effective. In contrast, only the Pre-Prep eradicated internal infection, but reduced laboratory germination. Other treatments reduced the Xcc levels, as measured by CFU/gram of seed. Continued studies were conducted in year two without the Oxidate treatment, and a proprietary modified hot water treatment was included (Seminis, Oxnard, CA, USA). Four new Xcc infested seed lots were tested: cabbage with low level of external infection, cabbage with low level of both internal and external infection, cauliflower (B. oleracea var. botrytis) with low level of internal infection and kohlrabi (B. oleracea var. gongylodes) with high level of both internal and external infection. All seed treatments eradicated Xcc from the three lots with either low internal, external or internal + external infection. The greatest challenge was the highly infected kohlrabi lot. Only the eXccit treatment was able to eradicate the external infection, while the HWT for 30 minutes, Pre-Prep and eXccit treatments eradicated the internal infection. It must be noted that several lots used in these studies may not pass suitability tests conducted by the seed enhancement companies.

SEED-BORN BARLEY STRIPE MOSAIC VIRUS IN EGYPT: INCIDENCE, EFFECT OF VIRUS AND SEED-TRANSMISIBILITY ZEIN, SALWA & ABOUL-ATA, ABOUL-ATA

Plant virus and Mycoplasma Res. Sec., Plant Pathol 30 D, El-Karama St., Alf Maskan, PO Box 11123, Cairo, Egypt, [email protected]

Generally, BSMV has low incidence in Egypt. It has highest incidence at Minia (7%) and Menufia (4.9%) in 2002-2003 growing season where certified seeds are not fully adopted. It had also low incidence throughout 1996 and 97 growing seasons. BSMV sometimes can cause plant death after having severe symptom appearance on few plants. Most plants can stay alive with severe virus effect. Germination in BSMV-diseased seeds has decreased by 9% to 43.8%. It causes decreasing in number of tillers and decreasing in plant height 22 Budapest, Hungary, May 17th – 19th, 2004

(16.5% - 55.3%) as well. It stops grain filling and sometimes spikes formation. It can stop seed formation i.e. Sids 7 wheat local variety. It decreases number of seeds / plant (28.2% - 90.1%) and weight of seeds / plant (20% - 94.1%) according to degree of severity of infection of which has high positive correlation with those previous factors. BSMV incidence is high in virus-diseased seeds using TBIA (27.8% - 70.0%). Certified seeds usage has to be adopted by the farmer to prevent BSMV distribution.

Keynote – Molecular Methods and the Future of Seed Testing By Gerry Saddler, United Kingdom

MOLECULAR METHODS AND THE FUTURE OF SEED TESTING MCEWAN, M., COCKERELL, V., MULHOLLAND, V., SOUTHWORTH, C., KERR, E., REID, A. & SADDLER, G.S.

Scottish Agricultural Science Agency (SASA), East Craigs, Edinburgh, EH12 8NJ, United Kingdom, [email protected]

A range of molecular methods have been, and are being developed at SASA for use in seed testing. Much of our work is focused on the development of seed health tests, however increasingly efforts are being directed towards methods appropriate for the detection of GM and varietal identification. Examples will be drawn from recent research into the development of a quantitative PCR assay for the fungal pathogens of wheat; Microdochium nivale and Tilletia tritici. In the case of the latter, quantification of these pathogens in seed lots has been established using both competitive PCR assays and fluorescent real-time PCR. Recent research into the development of genotyping methods for a range of crops will also be presented. In the majority of cases the use of fingerprinting methods based on microsatellites is the method of choice, however data will be presented that show that this approach is not without its problems in certain crops. The approaches adopted in establishing databases for the recognition of barley and pea varieties will be highlighted and the future direction of this work discussed. In addition, results will be presented on recent work into the detection and characterisation of GM contaminating events in seed lots, the type of sampling regimes and forensic approach employed.

SESSION 6 – Seed Improvement Chaired by Hugh Prichard, United Kingdom

THE USE OF RAPID AGEING AND CONTROLLED DETERIORATION TO EVALUATE IODINE VAPOUR TREATMENTS TO IMPROVE SEED STORAGE POTENTIAL. DE ATRIP, N., MATTHEWS,* S. & POWELL, ALISON A.

Department of Agriculture and Forestry, Hilton Campus, Block C, University of Aberdeen, Aberdeen, UK *Author for correspondence

Work on several crops (wheat, rice, mustard, mungbean, eggplant and radish) has shown that halogens such as iodine and bromine can reduce the rate of deterioration of seed stored for several months. The aim of the research reported was to investigate more rapid methods (days rather than months) for the evaluation of treatments to slow down deterioration, with a view to developing a potential screening technique to evaluate potential seed treatments. Oilseed rape seeds (cv Apex), harvested in 2000, were used. Both high vigour, unaged seeds and low vigour seeds (seeds aged for 12h at 20% moisture content and 45°C, followed by drying back) were exposed to iodine vapour (0, 8, 18 or 24h) in a transparent plastic desiccator placed in a glass fronted incubator at 25°C. The seeds were subsequently subjected to rapid ageing at 20% moisture content (mc) and 45°C for up to 36h, dried back and germinated at 20 ± 2°C in the dark for 14 days. All iodine treatments consistently 23 27th ISTA Congress – Seed Symposium improved the laboratory germination after rapid ageing. Initially unaged, high vigour seeds showed the greatest response and 18h was the most effective exposure time, maintaining 100% germination after 36h rapid ageing, compared with 62% for untreated seeds. The rate of germination after rapid ageing was consistently increased by the iodine treatment, most markedly so for the unaged seed after 36h rapid ageing. Assessment of the extent of deterioration in germinable seeds, using the CD vigour test (24h at 20% mc and 45°C), revealed that all iodine treatments improved germination after CD, especially for the unaged seeds. The clearest evidence of a reduction in the rate of deterioration was seen following the 18h iodine treatment and 24h rapid ageing, when the treated seed gave a CD germination of 83% compared with 13% in the untreated control. The role of halogens in retarding deterioration and the potential for further reducing the duration of the screening method will be discussed.

EFFECT OF SALT PRIMING ON SEEDLING EMERGENCE AND GROWTH OF WATERMELON SEEDS DURING DEVELOPMENT DEMIR, I. & MAVI, K.

Ankara University, Agriculture Faculty, Horticulture Department, 06110 Ankara/Turkey

The effect of salt-priming (2% KNO3, 20°C; 6 d) on emergence percentage, mean emergence time, seedling fresh weight and cotyledone length of watermelon (Citrillus lanatus (Thunb.) Matsum & Nakai) seed lots harvested 15, 20, 25, 35 and 45 daa (days after anthesis) from the plants grown in warmer and cooler regions in 2002 were investigated. Neither treated nor untreated watermelon seeds of 15 days grown in cooler region were able to emergence; however the same batch grown in warmer region had 25, 20% emergence in control and treated seeds, respectively. The major effect of the treatment was seen in earlier harvests (15, 20 daa) than later ones. Treated seeds showed 20% higher emergence, 3-4 days of shorter emergence time, 68 and 82 mg higher seedling fresh weight in seeds harvested 20 daa in both regions. In more mature lots of 25, 35 and 45 daa, treatment was less effective on seedling growth. While, mean emergence time of all seed lots harvested in both regions reduced significantly and being maximum of 4 days at 20 daa. The benefit gradually reduced, as seeds get more mature. Percentage of seeds germinated but not able to emerge on top of the soil varied between 0-6.7%, in treated but it was up to the 22.7% in control seeds which was varying among the lots. It can be concluded that salt priming can be useful for improving uniformity, seedling emergence and growth of heterogeneously matured watermelon lots particularly in early spring sowings.

EFFECTS OF SAND PRIMING ON GERMINATION, PHYSIOLOGICAL CHANGES AND FIELD PERFORMANCE IN DIRECT-SOWN RICE (ORYZA SATIVA L.) HU*, J., ZHU, Z.Y., SONG, W.J., WANG, J.C. & NAGANAGOUDA R.

Department of Agronomy, Zhejiang University, Hangzhou 310029, China ∗Fax: +86-571-86049815, [email protected]

A new priming method called sand priming was developed using sand as a priming solid matrix in present experiment. Seeds were mixed with sands that contained 3.8% (v/w) water and sealed in plastic box, and then were primed at 18 for 72 h. Four varieties of rice were used to investigate the effect of sand priming on germination and physiological changes of direct-sown rice (Oryza sativa L.) in the laboratory. The results showed that the energy of germination, germination percentage, germination index and vigor index were improved in four varieties. Meanwhile, seedling height, root length, number of root and root dry weight were significantly higher than the nonprimed controls. The activity of peroxidase (POD), catalase (CAT), α- and β-amylase were increased in four varieties after sand priming. Besides, the content of chlorophyll in leaves, soluble sugar and free proline in shoots all increased in sand primed seeds. Field experiments showed that the seed establishment and yield in sand primed seeds were significantly increased by 19.8% ~ 22.9% and yield by 9.8% ~ 31.2%, respectively as compared to soaked seeds without priming. It is indicated 24 Budapest, Hungary, May 17th – 19th, 2004 that sand priming method may help to improve seedling establishment in direct-sown rice and possible to be used in the field crop production.

SEED TESTING AND THE EFFECT OF INSECTICIDAL ACTIVE INGREDIENTS ON EMERGENCE OF HYBRID MAIZE SEED JONITZ, A. & LEIST, N.

LUFA Augustenberg, Neßlerstr. 23, 76227 Karlsruhe, Germany

Seed is the basic means of agricultural production and its quality is therefore of decisive importance for a farm’s success. Increasingly, seed is being treated with insecticidal active ingredients, which give seedlings effective protection against pathogenic organisms. Good phytotolerability is essential for seed treatments, as seedlings can be particularly sensitive. The insecticidal agents clothianidin, imidacloprid, and thiamethoxam, which belong to the new, neonicotinoid group of active ingredients, have different physicochemical properties and therefore differ in regard to their uptake by the plant. Their phytotolerability was tested under different environmental conditions in maize. The effects on emergence behaviour were investigated in relation to soil type and soil moisture content, in experiments carried out with five types of soil. Comparison of the three active ingredients in sandy loam at 40 % WHCmax showed differences in emergence depending on the active ingredients´ solubility in water. On the 4th day after sowing, the picture seen with clothianidin, the active ingredient with the lowest aqueous solubility, was the same as that observed in the control system. Treatment with imidacloprid delayed emergence only slightly, whereas treatment with thiamethoxam, the agent with the highest aqueous solubility, delayed emergence to marked extent. The active ingredients had greatest effects in the light soils. With increasing soil moisture content, the maize seedlings generally emerged more quickly and showed less effects than in dry soils. The observed effects on growth were clearly discernible, but within a few days the differences had been made up. Final emergence was thus not discernibly impaired.

EFFECTS OF DIFFERENT POLYMER COATING MATERIALS AND APPLICATIONS ON THE STORAGE LIFE AND AGEING OF ONION (ALLIUM CEPA L.CV. AKI) SEEDS: I. ASSESSMENT OF APPROPRIATE POLYMERS AND DOZES KAVAK, SULEYMAN & ESER, BENIAN

Ege Universty Faculty of Agriculture, Department of Horticulture, 35100 Bornova, Izmir, Turkey, [email protected], [email protected], [email protected]

Viability of seeds and storability are controlled by temperature and relative humidity during storage. A reduced rate of water uptake and water vapour movement into the seeds during storage could control by coating the seeds with hydrophobic polymers. Therefore, polymeric coatings allow the storage of the seeds required from harvest to sowing or planting. The objective of this study was to evaluate the effects of different hydrophilic and hydrophobic polymers coatings on the storage life and ageing of onion seeds. For this purpose Discoshine L-88 blue, Carboxymethylcellulose, Daran SL112, Daran 8600 C, Linseed oil, polyethylene and vinamul 3240 polymers were used. According to the experimental results, hydrophobic polymer coatings did not retard water vapour at high RH conditions for onion seeds as expected. Some of these polymers negatively affect germination as compare to uncoated seeds. These polymers possibly form a hard film that could negatively affect germination and could be a toxic effect on the seeds. After 10 weeks of storage period, moisture contents of coated seeds were not different from uncoated seeds and polymer coating of onion seeds caused a physical barrier to the radicle growth.

25 27th ISTA Congress – Seed Symposium

SULPHURIC ACID SCARIFICATION EFFECTS ON BRACHIARIA BRIZANTHA, B. HUMIDICOLA AND PANICUM MAXIMUM SEED DORMANCY RELEASING USBERTI, ROBERTO & MARTINS, LEILA

Plant Protection Agency, Caixa Postal 960, CEP 13073-001, Campinas, Brazil, [email protected]

Forage grasses present seed dormancy, which reduce percentages in germination test as compare to viability results obtained by tetrazolium test. The objective of this work was to evaluate the effect of H2SO4 scarification on seed dormancy releasing in 630, 94 and 82 samples of B. brizantha, B. humidicola and P. maximum, respectively, tested in Central Seed Testing Laboratory, Campinas, Brazil, from 1991 to 1999. Germination tests used two 4x100 intact and scarified seeds (15’, 10’, 5’, respectively). Mean germination (T50) and variability among germination replicates were also analysed. Statistical analysis was performed by t- test paired samples for means. Scarification promoted reductions in T50, while variability among replicates was reduced in B. brizantha. Scarification increased germination in B. brizantha and P. maximum, but was deleterious in B. humidicola.

SESSION 7 – Physiological Basis of Seed Quality Chaired by Françoise Corineau, France

SEED LONGEVITY CHART AND MODELING TO PREDICT VIABILITY DURING OPEN STORAGE ANDREOLI, CLAUDINEI

Embrapa Soja, Brazil, de Andrade, Ramiro Vilela, Embrapa Milho e Sorgo, Brazil, [email protected]

The simplified equation Vt = Vi – tgβ.p has been incorporated into a seed viability chart for corn (Zea mays L.) in order to predicting the percentage of viability of any corn seed lot after any period of time under uncontrolled storage conditions. The objective of this work was to set a practical seed longevity chart to predict viability of corn seed lot during storage. From data of five corn seed lots stored at Sete Lagoas, MG, the value of the storage condition (σ) was calculated. This value (σ = 120 days) corresponded exactly to the time taken in days for the germination of corn seed lots stored at Embrapa, Sete Lagoas, State of Minas Gerais to fall to a certain value observed in the accelerated aging test. Once the storage condition is estimated, the viability chart can be constructed, which illustrates the relationship between initial germination, seed deterioration rate and storage condition over time. The chart can be used by any seed producer once the value of storage condition (σ) in situ is determined. Applications of the seed viability chart to short-term seed storage seem to be promising for seed growers. This predictive model is the first attempt of incorporating the effect of seed quality, actual temperature and relative humidity into a seed longevity chart of an open storage.

LIPID PEROXIDATION AND ACTIVITY OF SUPEROXIDE DISMUTASE ASSOCIATED WITH NATURAL AGING OF OIL MAIZE SEED BALEŠEVIĆ-TUBIĆ, SVETLANA, TATIĆ, M., MILADINOVIĆ1, JEGOR & MALENČIĆ2, Đ.

1Institute of Field and Vegetable Crops, Novi Sad, Maksima Gorkog 30, 21000 Novi Sad, Serbia and Montenegro 2Faculty of Agriculture, Novi Sad, Trg D. Obradovića 8, 21000 Novi Sad, Serbia and Montenegro

Biochemical changes that occur in the seed during natural aging are very significant for seed quality and longevity. Changes in lipid content and composition, which mostly occur due to lipid peroxidation, can have a significant negative effect on the properties of stored seed. This study was conducted to determine effect of natural aging on changes in the fatty acid content, germinability, lipid peroxidation and activity of superoxide dismutase (SOD) in 26 Budapest, Hungary, May 17th – 19th, 2004 seeds of two hybrids and three lines of oil maize. The results indicate that natural aging decreased seed germination and caused changes in the fatty acid content. The increase of lipid peroxidation occurred with increased duration of aging. During process of natural aging some decrease in SOD activity were noticed. These data suggest that the peroxidative changes in the lipids may be linked to the reduced germination and deterioration of oil maize seed.

THE EFFECT OF DIFFERENT POST-HARVEST DRYING METHODS ON SEED QUALITY FROM GREEN (IMMATURE) AND RED (MATURE) BERRIES OF WOODY NIGHTSHADE (SOLANUM DULCAMARA L.) CONEYBEER, JULIA, ADAMS, JOHN & PROBERT, ROBIN J.

Seed Conservation Department, Royal Botanic Gardens, Kew, Wakehurst Place, Ardingly, West Sussex, RH17 6TN, UK, [email protected]

Green and red berries of woody nightshade were harvested on a single day. Berries were then either held under ventilated ambient conditions for 21 d prior to seed extraction or, seeds were extracted on the day of harvest and divided into three different drying methods. 1) seeds held under ambient conditions for 21 d as above. 2) seeds dried immediately under standard seed bank conditions for seeds of fleshy fruits (open drying 15°C and 15% RH). 3) As 2 except seeds held in a ventilated box designed to slow down the rate of drying. Immediate, standard drying of seeds extracted from green berries results in low seed viability (~40%) and poor storage potential. Seed quality was improved if seeds were dried more slowly in a ventilated box but highest seed quality was obtained when green berries were allowed to mature for 21 d under ambient conditions prior to seed extraction and drying. Seed from mature red berries behaved differently. Seeds from berries held under ambient conditions for 21 d were of poorer quality compared with seeds extracted on the day of harvest and dried immediately. However, slow drying of seeds at 15°C in a ventilated box still resulted in higher seed quality compared to seed dried under standard seed bank conditions.

GENETIC DISSECTION OF MAIZE RESPONSE TO A DEFOLIATION TREATMENT DURING MATURATION INDUCING TOLERANCE TO COLD AT GERMINATION. FRASCAROLI, E. CASARINI, E. & CONTI, S.

University of Bologna, Dept. of Agroecological Sciences and Technology, Viale Fanin 40, 40127 Bologna, Italy, [email protected]

It has been proved that defoliation of mother plant during kernel maturation can improve cold tolerance of germinating seeds and of embryos rescued few days after defoliation. Objectives of this study were to evaluate a mapping population segregating for tolerance to cold during seed germination, and for reaction to defoliation, to detect and characterize quantitative trait loci (QTLs) controlling cold tolerance and its acquisition during defoliation. One-hundred-seventy-one families of the population derived from the cross Lo1016 x Lo964, previously genotyped at 161 marker loci, were selfed and seeds were harvested from defoliated (D) or non-defoliated (ND) mother plants. Composite interval mapping was used to identify QTLs for germination at 25°C (G25) and 9°C (G9), and for reaction to defoliation. In the analysis on ND plants, five QTLs were detected for G25 and eight for G9, explaining respectively 31 and 41 % of the variation. Four QTLs were in common, while four were involved in G9 only. Four QTLs were detected in defoliated plants analysis for G25 and five for G9, explaining 17 and 26 % of the variation, respectively. Only one was in common. QTLs controlling germination can be used for marker-assisted selection to improve this trait. The presence of genomic regions possibly involved in reaction to defoliation and its relation with cold tolerance is also discussed. Results reported in this study indicated that reaction to defoliation could be used as a model for better understanding genetic control and physiological mechanisms at the basis of maize cold tolerance at germination.

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EFFECT OF HIGH TEMPERATURE STRESS DURING SOYBEAN SEED DEVELOPMENT ON GERMINATION AND VIGOR TEKRONY, DENNIS M., EGLI, DENNIS B. & SPEARS, JAN

Dept. of Agronomy, University of Kentucky, 429 Plant Science Bldg, Lexington, KY 40546-0312, United States, [email protected]

Several environmental factors occurring during soybean seed production have been reported to influence seed quality. This investigation evalulated the effect of high temperature during seed development and maturation on the germination and vigor of seed produced in the phytotron (1998, 2000) and the field (2000 to 2002). Cultivars of variable maturity (‘Hutcheson’, ‘McCall’) were grown in the phytotron at temperatures of 27/22 (control), 33/22 and 38/27) and in the field (‘Hutcheson’, and ‘DP4690RR’) at four locations (Kentucky, Mississippi, Arkansas and Texas)in the USA. Seeds were hand harvested at maturity (brown pods), hand threshed and all shriveled and abnormal seeds were removed before testing for germination and vigor (accelerated-aging germination, conductivity). Maximum daily phytotron temperatures of 33 or 38 °C during seed development reduced seed size, increased shriveled seeds and lowered seed quality. Standard germination of normal seeds (no visual abnormalities) developing at 33°C was 98%, while seed developing at 38°C were 3 to 72 %. Seed vigor was significantly (P > 0.05) reduced at 33 °C and unacceptable at 38°C. Average maximum field temperatures during seed filling (growth stage R5 to R7) ranged from 24.0 (Kentucky) to 37.6ºC(Texas). When all seed lots infected with Phomopsis longicolla (Hobbs) were removed from the analysis there was a significant decrease in standard germination and accelerated-aging germination as mean maximum temperatures during seed filling increased for Hutcheson, but not for DP4690RR. Standard germination declined linearly (r2 = 0.49) from near 100% at 24ºC to 85% at 36ºC, while the decrease in accelerated-aging germination was curvilinear (R2 = 0.88) and reached 9% at 36ºC. Seed vigor (accelerated-aging germination) was much more sensitive to high temperature stress than standard germination. There were also cultivar differences with seeds of Hutcheson more sensitive to high temperatures than DP4690RR or McCall. Our findings in the phytotron and field demonstrate that high temperatures during seed filling, in the absence of seed infection with Phomopsis longicolla (Hobbs) and physical injury, reduced soybean seed germination and vigor.

100-SEED TEST FOR DESICCATION TOLERANCE AND GERMINATION: A CASE STUDY ON EIGHT TROPICAL PALM SPECIES WOOD, CHRISTOPHER B., HODGES, SIMON, VAUTIER, HELEN J. & PRITCHARD, HUGH W.

Seed Conservation Department, Royal Botanic Gardens Kew, Wakehurst Place, Ardingly, West Sussex RH17 6TN, UK, [email protected]

A test was devised for seed desiccation tolerance, the crucial first step in the ex situ preservation of germplasm and its sustainable use. Its unique attribute is the maximum characterisation of seed lots using the minimum number of seeds. Data recorded includes seed dimensions, fresh and dry weight, moisture content, relative humidity, germination total and rate in one environment before and after desiccation with silica gel. Seeds of eight tropical palms from the genera Phoenix and Syagrus were studied. Six were found to be desiccation tolerant, a trait that closely matched the species natural habitat; two species, Phoenix roebelenii and Syagrus schizophylla, are provisionally classified as having desiccation sensitive, Type III seeds. In general, germination was observed to be rapid, being complete in one to two months. Two species exhibited an increase in germination during short-term (weeks) moist storage, probably indicative of a progression in embryo development. The results show that the 100-seed test can be used as an efficient and effective first screen for potential storage of seeds from diverse species and can support the rapid production of seed biology checklists for species conservation and sustainable use.

28 Budapest, Hungary, May 17th – 19th, 2004

Poster Sessions Chaired by Zita Ripka, Hungary

1. Application of Advanced Technologies Topics: Identification of GM; varietal identification; diagnostics in plant pathology

1 - COMPARING TECHNIQUES TO DETECT SOYBEAN SEEDS TOLERANT TO GLYPHOSATE ANDRE TILLMANN¹, MARIA ANGELA, DOS SANTOS MADRUGA CUNHA2, CRISTINA CLAUDETE, FUNGUETTO2, IZABEL, BICCA DODE3, LUCIANNA & AMARAL VILLELA¹, FRANCISCO

¹bolsista CNPq, UFPel/FAEM, Caixa Postal 354, CEP 96001-970, Pelotas-RS, [email protected] 2UFPel/FAEM, Pelotas-RS 3UCPEL, Pelotas-RS

There is a worldwide effort to establish methods for the detection of GMOs, bearing in mind the importance of international commercialization. Recently, bioassays have been carried out to evaluate plant phenotype, such as ELISA tests and kits to detect specific proteins expressed from transgenic DNA. Using PCR it is also possible to detect specific DNA sequences. The aim of this research was to compare the efficiency of the methods used to detect Roundup Ready® (RR) soybean and to characterize the seedlings response to the herbicide. Soybean seeds tolerant to glyphosate were submitted to various bioassays (pre- imbibed, imbibed, and immersed in glyphosate solution), seedlings sprayed with glyphosate, kit Trait Test and the PCR method. The results indicated that the methods are efficient in detecting the RR seeds; the bioassays were the most efficient, considering the cost benefit ratio. The non-GMO seedlings presented with some abnormalities: areas of thickness, longitudinal striations, yellowing of the hypocotil, inhibition of primary root development, secondary root absence and reduced length. The radicular system was the most affected, compared to the aerial growth.

2 - EXAMPLES OF THE STATISTICS COMMITTEE’S RECENT WORK BARABÁS1, JULIA & GREGOIRE2, SYLVAIN

1Budapest University of Economic Science and Public Administration, Hungary, 1118. Villányi út 35, Mathematics and Infomatics Dep., Hungary, [email protected] 2GEVES, France

The ISTA Statistical Committee helps to solve different seed testing problem of the difference committee of ISTA. The main goals are to give both practical and methodology advises to the colleagues. This poster illustrates through examples these cooperation from the past 3 years and shows that the demands were coming from users. The Statistical Committee tried to provide solid and appropriate mathematical background. The examples are: - GMO's sampling plan, proficiency testing, error rates - purity of seed mixtures: new methodes for composition check and chack the labelled value -worked out of Statistical analysis of Plant Disease Committee method validation programs -Give information and free tools on the ISTA web-site

29 27th ISTA Congress – Seed Symposium

3 - EVALUATION OF IMAGE ANALYSIS IN DETERMINING THE RELATIONSHIP BETWEEN MECHANICAL DAMAGE AND SEED VIGOR IN CORN CICERO, SILVIO MOURE

São Paulo University, Banzatto Junior, Hodair Luiz, São Paulo University, [email protected]

Image analysis is a very promising technique to determine mechanical damage in seeds. This precise method examines seeds individually using enlarged images in which damaged areas as well as their exact location and extension can be found and examined in detail. Since this is a non-destructive method, analyzed seeds can be submitted to physiological tests to establish the relationship between mechanical damage and quality loss. The objective of the present paper was to study the effects of mechanical damage on corn seed vigor using image analysis. Fifty seeds from CO 32, AG 6690 and Attack cultivars were visually selected to form a sample of whole seeds with varying degrees of mechanical damage. Seeds were X-rayed, photographed (ventral and dorsal sides) and submitted to a cold test. Photographs were repeated after the cold test. Images were transferred to a computer so that pre and post cold test images could be examined simultaneously on the monitor to determine possible relationship between cause and effect. Results indicate that the method under study permits association of mechanical damage with eventual losses caused to corn seed vigor.

4 - APPLICATION OF IMAGE ANALYSIS TO STUDY SEED IMBIBITION UNDER SEVERAL ENVIRONMENTAL CONDITIONS DELL’AQUILA, ANTONIO

Insitute of Plant Genetics, CNR, 70126 Bari, Italy

Most information on seed germination and physical seed swelling during imbibition is based on the subjective evaluation of ‘visible germination’ and gravimetric assessment, which are generally known to be sources of error. Computer-aided image analysis represents a new approach to monitor the imbibition process based on the automation and the measurement of seed size parameters. The technique has been applied to study: (a), the changes of cabbage (Brassica oleracea L.) seed size parameters, such as area, perimeter, width and length, during the first and second phase of water uptake; (b), the swelling of deteriorated cabbage seeds exposed to 0.44M NaCl for brief period of time before radicle emergence, by measuring variations of the seed area during salt imbibition and after stress removal; (c), the relationships between area and roundness factor, as parameter of seed shape change, measurements and early radicle growth in cabbage, lentil (Lens culinaris Medick.), pepper (Capsicum annuum L.) and tomato ( Lycoperiscon esculentum L.) seeds during the third phase of imbibition curve; (d), a more accurate assessment of the germination parameters in cabbage and radish (Raphanus sativus L.) seeds imbibed under different temperature conditions. The results show that the seed area may be taken as a good marker of imbibition process during the Phase I and II of water uptake, while the timing of germination onset and radicle elongation may be better determined by the roundness factor. In addition, the rapid image processing by computer has allowed the creation of a database of germinating seed images for several species, available on the web site: http://germimaging.ba.cnr.it .

30 Budapest, Hungary, May 17th – 19th, 2004

5 - USING ARTIFICIAL VISION TO MONITOR GERMINATION TIME COURSE OF SUNFLOWER (HELIANTHUS ANNUUS L.) SEEDS DUCOURNAU1, S., FEUTRY2, A., PLAINCHAULT3, P., REVOLLON4, P., VIGOUROUX5, B. &. WAGNER1, M.H

1Station Nationale d’Essais de Semences (GEVES-SNES), rue G. Morel, B.P. 24, 49 071 BEAUCOUZE CEDEX, France, [email protected], [email protected] 2Laboratoire de Technologie des Semences, VILMORIN, 49250, LA MENITRE, France, [email protected] 3C.E.R., E.S.E.O., 6 rue Merlet de la Boulaye, B.P. 30926, 49009 ANGERS CEDEX, France, [email protected] 4Institut National d’Horticulture, 2 rue Lenôtre, 49045 ANGERS CEDEX 01, France, [email protected] 5Laboratoire d’Ingénierie des Systèmes Automatisés (LISA, CNRS FRE 2656 ), Institut Universitaire de Technologie, BP 42018, 49016 ANGERS CEDEX, France, [email protected]

A computer-controlled system was used to monitor the germination time course of sunflower seeds. The system integrates a Jacobsen table, a controlled lighting environment, a video camera and a computer. Software was developed to control image recording and image analysis. The algorithm retained for detecting and counting the germinated seeds was validated in two ways: 1. First, human operators read images independently and the variability between readers was compared to the computer reading. 2. Then, computer counts were compared to counts of germination carried out directly on the Jacobsen table. The whole equipment (automatic shooting and algorithm connected to the germination system) was tested to plot germination time courses of three sunflower seed lots at 20°C. Detailed germination curves were obtained allowing a perfect fitting in a probit model and a comparison of the seed lots. These results confirm the high potential of artificial vision in quality evaluation of seeds.

6 - MORPHOLOGICAL AND CHEMICAL IDENTIFICATION OF NEW VARIETIES OF SOME PEANUT (ARACHIS HYPOGAEA L.) EL-BAGOURY, O.H.1; EL-EMERY2; M.I; EL HABBAL, M.S.1; AND NAGUIB, N.A.2

1Agron. Dept., Faculty of Agriculture, Ain Shams University. Cairo, Egypt. 2Seed Tech. Dept., Field Crops Res. Inst. Agricultural Res. Center, Giza, Egypt.

This study was carried out with the objectives of determining the actual differences between various peanut genotypes under test at different growth stages. The necessity for such information was to assist in the quality control and certification tests. Three peanut (Arachis hypogaea L.) genotypes (Giza 4, Giza 5 and Hybrid 8), were selected and certain selected quantitative and qualitative characters were studied. Certain quantitative and qualitative morphological characters were investigated on seed, seedling and adult plants. The results revealed great differences in certain morphological characters and chemical composition including crude protein, total carbohydrates, crude oil and fatty acids were tested in seeds. Moreover, the patterns of seed protein by using SDS–PAGE. It was found that each genotype was characterized by proteins with specific molecular weight. Therefore, electrophoretic analysis is an important tool for the identification of cultivars.

31 27th ISTA Congress – Seed Symposium

7 - MOLECULAR DETECTION AND IDENTIFICATION OF BACTERIAL DISEASES IN SEED POTATOES FEKETOVA, MIROSLAVA & HORVATH, LUBOMIR

Department of Molecular Biology, Central Control and Testing Institute for Agriculture, Hanulova 9/A, 841 01 Bratislava, Slovak Republic, tel./fax: +421 2 64462089, [email protected]

Bacteria Ralstonia solanacearum (RS) and Clavibacter michigenensis subsp. sepedonicus (CMS), which cause very dangerous bacterial diseases “potato brown rot” and “potato ring rot”, belong to the quarantine organisms in Slovakia, because their occurence have not been detected in domestic production. In the frame of state research project (1999-2002) carried out development and standardisation of DNA-based testing procedures for detection and identification bacteria RS and CMS in seed potatoes and ware potatoes by using of polymerase chain reaction (PCR). Ralstonia solanacearum was detected by using simple PCR according to EC directive 98/57/EC with specific primers OLI-1 and Y-2, that give 288 bp amplicon and with additional primers derived from DNA encoded 16SrRNA. Positive PCR fragments are confirmed by RFLP analysis with AvaII. Clavibacter michigenensis subsp. sepedonicus was detected by using simple and preferably multiplex PCR with specific primer pairs based on region 16S-23SrRNA genes. It was indicated as very useful a cultivation of bacteria in the liquid or solid growth media before testing especially in cases of weak bacterial infections. Standardized PCR tests are performed in the official phytosanitary diagnostics for verification of positive immunofluorescence and biological tests and for autonomous monitoring of presence of bacteria to prevent of undesirable spreading of mentioned quarantine bacterial diseases.

8 - STANDARDISATION OF DNA-BASED TECHNIQUES FOR DETECTION OF GMOS IN OFFICIAL TESTING HORVATH, LUBOMIR & FEKETOVA, MIROSLAVA

Department of Molecular Biology, Central Control and Testing Institute for Agriculture, Hanulova 9/A, 841 01 Bratislava, Slovak Republic, tel./fax: +421 2 64462089, [email protected]

The area of molecular detection and identification of genetically modified plants (GMOs) based on the detection of DNAs of non-plant origin, is highly actual in the sphere of seed testing, variety testing, GMOs labeling and coexistence of GMO crops with conventional and organic farming. In the frame of the state research project (1999-2002) carried out development, verification and standardisation of testing procedures for detection of GMOs in plant varieties, seeds and mercantile by using polymerase chain reaction (PCR). PCR tests were performed for various products for the presence of glyphosate, Bt and glufosinate tolerant GMOs in soybean, maize, rapeseed, sugar beet, sunflower and other species, that are based in detection of 35S CaMV promotor, NOS terminator and specific DNA sequences of transgenic plants. In order to create compatibility with EU, screening and specific simple and multiplex PCR tests, including the methods of JRC EC Ispra, German Food Act, Swiss method and their modification with different primers, types of DNA extraction and purification were compared. For determination of relative contents of GMOs in the grain samples, semiquantitative test based on subsampling and sequence tests was successfully employed. On the ground of obtained results standard procedures for qualitative and semiquantitative detection of GMOs in agricultural testing in Slovakia were established. Optimal sensitivity of the methods was specified to 0,1% of GMOs contents. Positive results are compared to GMO standards (IRMM/Fluka), the specificity of PCR test is checked by set of positive and negative controls and positive PCR fragments are confirmed by the RLFP analysis. The contamination of samples by CaMV virus, that causes false positive results through detection of CaMV 35S promotor, is performed by the PCR detection of reverse transcriptase gene of this virus.

32 Budapest, Hungary, May 17th – 19th, 2004

9 - GENOTYPIC RESPONSE OF CHILLI FOR SEED YIELD AND QUALITY UNDER DIVERSED ENVIRONMENTAL CONDITION HUNJE, RAVI., SHEKHARGOUDA, M. & VYAKARANAHAL, B.S

Seed Research Laboratory, National Seed Project, University of Agricultural Sciences, Dharwad, Karnataka-India

Field and laboratory experiments were conducted over two year under three agroclimatic zones of Karnataka State-India on two premier local varieties viz. Byadagi kaddi and Dyavanur local chilli. Byadagi kaddi variety recorded significantly higher growth and yield parameters compared to Dyavanur local. Byadagi kaddi variety produced higher fruit length (15.86 cm), fruits per plant (109.50), higher fruit yield per plant (81.57 g) and per ha (1318 kg) and seed yield (498.42 kg/ha) and also recorded higher root length (9.81 cm), shoot length (9.03 cm) and vigour index (1694) compared to Dyavanur local. Dharwad location recorded higher number of fruits per plant (109.63), fruit weight (1.33 g), fruit yield per plant (86.33 g) and per ha (1468 kg) and seed yield (512.42 kg/ha) over Hanumanmatti and Annigeri location and seed quality parameters also showed similar trend. Second and first picking recorded higher fruit length (13.70 and 12.52 cm) and fruit per plant (28.94 and 35.63), fruit yield (428.56 and 380.40 kg/ha), seed yield (159.61 and 140.85 kg/ha) and also higher seed quality parameters viz., germination (88.44-80.06) field emergence (83.04-80.69), vigour index (1311-1146) and lower in electrical conductivity (1.47-1.56 dSm-1) of seed leachates respectively. Byadagi kaddi variety at Dharwad location performed better for seed yield and quality parameters.

10 - LONG TERM PRESERVATION OF CHILLI SEEDS USING CHEMICALS & BOTANICALS HUNJE, RAVI., SHEKHARGOUDA, M. & VYAKARANAHAL, B.S

Seed Research Laboratory, National Seed Project, University of Agricultural Sciences, Dharwad, Karnataka-India

Laboratory experiments were conducted to study the influence of seed treatments with chemicals and bio-products on two chilli varieties viz., Byadagi kaddi and Dyavanur local at the Seed Research Laboratory, National Seed project, University of Agricultural Sciences, Dharwad, Karnataka, India for 20 months period. The chilli seeds deteriorate at a rate sufficiently rapid to make them a poor planting material leading to poor germination, slow emergence, weak growth, inadequate field stand etc, all orienting towards reduced yield. By dressing with fungicides, plant products and seed invigouration result in improved seed quality for two-three planting reasons. The chilli seed germination can be maintained above minimum seed certification standard (60%) for a period of 12 months under ambient condition. Among the varieties Byadagi kaddi retained better seed quality parameters compared to Dyavanur local. The seeds treated with potassium iodide (10-3M), calcium oxy-chloride (4 g/kg) and Pongamia leaf powder (40 g/kg) recorded significantly higher seed quality parameters and maintained the seed germination above minimum seed certification standard (60%) over a period of 20 months under ambient conditions of storage.

11 - STUDY FOR GM SOYBEAN ANALYSIS USING ELISA JUNG, JIN-KEE

Experiment Research Institute of National Agricultural products Quality Management Service(NAQS), 560, 3-Ga, Dangsan-Dong, Youngdeungpo-Gu, Seoul, Republic of Korea, ([email protected])

The study for GMO analysis is progressing actively according to development of GM corps in worldwide. There are two methods of PCR and immunoassay for GMO analysis. For the

33 27th ISTA Congress – Seed Symposium purpose of effective GMO analysis we made comparative experiment by ELISA analysis according to content of proteins in each nation's soybeans, levels of grinding, methods of sample preparation, sorts of standard materials and percentage of GMO contents using ELISA analysis. Mixtures of RR soybean and non GM Korea cultivar soybean were used in this study. Protein contents in non GM soybean of Korea, China and America and RR soybean were compared. And also protein contents of standard materials grinded as 100mesh and sample grinded as 40mesh are compared. We made comparative experiment of GM soybean of 3% which is Korea's GMO threshold using SDI ELISA kit according to methods of preparation. Difference of GMO contents between IRMM and SDI standard materials was investigated. 0.5, 1, 3, 5, 10, 20, 40, 60, 80, 100% GM soybean prepared in our laboratory were analysed using SDI ELISA kit. As the result, content of proteins were various according to origin and varieties as Korea soybean is 14.2 ug/ul, China soybean is 16.3ug/ul, America soybean is 10.2 ug/ul and RRS is 11.4 ug/ul. Protein contents of GM soybean grinded as 100mesh and 40 mesh US screen were 17.2 ug/ul and 16.2 ug/ul respectively in average so it makes no difference in ELISA analysis. GMO analysis of 3% GM soybeans made by powder mixed after passed through 40 mesh US screen showed more stable results than powder passed through 40 mesh US screen after mixed as karnels as the former is 2.9-3.0% and the later is 3.1-3.9%. ELISA analysis results between 3% GM prepared with IRMM and SDI standard materials showed similar results of 2.9-3.8% and 3.0-3.9% respectively. ELISA analysis of each percentage of GM soybean (0.5, 1, 2, 3, 5, 10, 20, 40, 60, 80, 100%) made by RR soybean and non GM Korea cultivar soybean showed higher coefficient of variation (CV) in lower ratio of GMO than higher ratio of GMO as CV of 0.5%, 1% GMO is 12.7, 7.9% and CV of 100%, 80% GMO is 7.4%, 3.3% respectively. It showed ELISA analysis of higher ratio of GMO contents is more stable results than lower ratio.

12 - IDENTIFICATION OF THE STEM LUCERNE NEMATODE DITYLENCHUS DIPSACI BY PCR KERKOUD1, M., GUIMIER1, C., GUENARD1, M., LECHAPPE1, J & ESQUIBET2, M.

1GEVES-SNES, rue Georges Morel, 49071 Beaucouzé Cedex 2INRA de Rennes, BP 35327, 35653 Le Rheu Cedex

Ditylenchus dipsaci is a quarantine phytoparasite composed of two races (normal and giant) which are difficult to distinguish from other Ditylenchus species. Using healthy seed constitutes the best way to avoid the dispersion of that nematode. Faced to the complexity of identification, the molecular tool can be one alternative to characterize and identify such a parasite. A technique based on polymerase chain reaction (PCR) was developed for the identification of D. dipsaci using nucleotide sequence information of the ribosomal DNA region. The internal transcribed spacer region ITS1 and ITS2, the gene 5.8S and partial genes 18S and 26S of two populations of the stem and bulb nematode D. dipsaci and phylogenetically related species were amplified with two universal rADN1 and rADN2 primers, sequenced, and digested with five restriction enzymes. The alignment of nucleotide sequences and analyses of digestion patterns indicated constant polymorphisms between D. dipsaci and related species which overlapped a TspEI restriction site. Two oligonucleotide primers set were designed for identifying this variable region. The first primer set (DdpS1 and rADN2) was specific to D. dipsaci when it was checked against nematode genomic DNAs of related nematodes. The second primer set (DdpS2 and rADN2) was specific to normal race of D. dipsaci. The first primer set is a good candidate for a species-specific reagent in a procedure for detection and identification of D. dipsaci by PCR. The results of this study will allow the development of a fast PCR test for routine detection of D. dipsaci.

34 Budapest, Hungary, May 17th – 19th, 2004

13 - IDENTIFICATION OF GROUNDNUT [ARACHIS HYPOGEA (L.)] CULTIVARS USING MORPHOLOGICAL TRAITS, CHEMICAL TESTS AND ELECTROPHORESIS KESHAVULU1, K., FARZANA2, JABEEN SAMBASIVA RAO1, P., RADHIKA1, K., MURALIMOHAN, REDDY B. & TONAPI, V.A.

1National Seed Project and Department of Seed Science and Technology, Acharya N G Ranga Agricultural University, Rajendranagar, Hyderabad, Andhra Pradesh 500 030, India 2Department of Genetics and Plant Breeding, Acharya N G Ranga Agricultural University, Rajendranagar, Hyderabad, Andhra Pradesh 500 030, India [email protected], [email protected]

Groundnut (Arachis hypogea L.) is one of the major oilseed crops in India, being grown across all the seasons. Multiplicity of genotypes and non availability of documented information on diagnostic characteristics of the groundnut varieties has complicated the issue of maintenance of genetic and varietal purity. To aid the process of distinctiveness, uniformity and stability, 87 cultivars actively grown for seed across the country were characterized and documented both morphologically and through chemical tests including electrophoresis at National Seed Project at Hyderabad from 1998 to 2002. The stable diagnostics characteristics through seed, pod and plant morphological characters from sowing to maturity, standardized chemical tests and SDS-PAGE electrophoresis of soluble seed proteins were used for cultivar identification. Most of the varieties showed variation in morphological characteristics, but few varieties did not exhibit morphological variability, and it was not possible to strictly assign them to a specific group. Different groupings for majority of the cultivars were possible by chemical tests. The banding pattern of soluble proteins by PAGE was significantly different in most of the varieties and specific groupings for cultivar identification were evident. Thus, the study clearly revealed the clear possibility of identifying and distinguishing groundnut cultivars based on combination of morphological characters, seed and seedling reaction to KOH, NaOH, 2,4-D and GA3 tests, and electrophoresis. This characterization is expected to help the Indian seed program in certification, quality control and grow-out testing of groundnut.

14 - IDENTIFICATION OF RICE CULTIVAR USING MORPHOLOGICAL, CHEMICAL TESTS AND ELCTROPHORESIS KESHAVULU1, K., SAMBASIVA RAO1, P., FARZANA2, JABEEN, & MANOHAR REDDY, N.

1National Seed Project and Department of Seed Science and Technology, Acharya N G Ranga Agricultural University, Rajendranagar, Hyderabad, Andhra Pradesh 500 030, India, [email protected] 2Department of Genetics and Plant Breeding, Acharya N G Ranga Agricultural University, Rajendranagar, Hyderabad, Andhra Pradesh 500 030, India

Rice (Oryza sativa L.) is one of the most important staple food of the world. It has been estimated that rice production in India must be doubled by the year 2025 to meet the requirements of population. Varietal identification has attained critical importance world wide in view of plant variety protection and maintenance of genetic purity in seed programmes. Due to proliferation of varieties, identification and maintenance of genetic purity has become complicated. One hundred and twenty four rice cultivars including varieties, parental lines and hybrids grown in Southern India were studied from 1999-2002. With a view of developing a scheme for variety identification using 58 morphological characters, seed and seedling reactions to chemical tests such as phenol, modified phenol, KOH, NaOH, 2, 4-D and GA3 including PAGE of soluble seed proteins. These rice cultivars showed variations for most of the characters from coleoptile emergence to seed maturity encompassing plant, leaf, stem, inflorescence, panicle and seed as qualitative and quantitative characteristics of each. The coleoptile colour and shape of legule, stem anthocyanin coloration of nodes, color colour and anthocyanin coloration of keel did not exhibit variability among cultivers. With the help of morphological characters, it was not possible to differentiate all the cultivars especially parental lines of hybrids. Different groupings were possible by chemical tests, however, complete identification of all the cultivars was not possible. The soluble seed protein profile

35 27th ISTA Congress – Seed Symposium by PAGE was quiet different for the parental lines and hybrids. Therefore, combined utility of morpho, chemical and electrophoretic techniques could help differentiating all the cultivars in an integrated manner to aid in maining better genetic purity and cultivar identity.

15 - NONDESTRUCTIVE CLASSIFICATION OF VIABLE AND NONVIABLE RADISH (RAPHANUS SATIVUS L.) SEEDS USING SINGLE SEED NEAR INFRARED SPECTROSCOPY MIN, TAI-GI & KANG, WOO-SIK

Daegu University, Korea, College of Natural Resources, Daegu University, Jinryang-eub, Kyungsan-si, 712-714 Republic of Korea, [email protected]

Near infrared (NIR) spectroscopy was used to classify viable and nonviable radish (Raphanus sativus L.) seeds. The spectra collected from single seed were perfectly distinguished from viable and nonviable seeds by principle component analysis (PCA). To discriminate viable and nonviable seeds, multivariate classification model was developed with a discriminant partial least squares (PLS) 2 method. The calibration model derived by PLS 2 method based on the first derivative NIR reflectance spectra in the range of 1100-2500 nm resulted in a 100% classification rate. The calibration model with the first derivative spectra predicted 94 % viable and 95% nonviable seeds from the unknown samples. The result shows the possibility of using NIR spectroscopy in a seed separating process in the future provided that appropriate sorting devices are developed.

16 - EFFECT OF SOYBEAN CULTIVAR AND SEED QUALITY ON CARBOHYDRATE AND ORGANIC ACID PHYTHIUM DISEASE DEVELOPMENT NANAYAKKARA1, R., ROTHROCK2, C.S. & RUPE2, J.C.

1Seed Testing Laboratory, Aluttarama, D.E. Longer, Department of Crop, Soil, & Environmental Sciences, Fayetteville, Arkansas 72701, Sri Lanka 2Department of Plant Pathology, Fayetteville, Arkansas 72701, Sri Lanka

Compounds released by germinating seeds are an important source of nutrients for soil microorganisms and are critical for soilborne pathogens to detect a host. Few studies have focused on the composition of seed exudates in relation to cultivar and seed quality and the role of these factors in resistance to infection by Pythium spp. and disease development. The objective of this research was to quantify and identify compounds in seed exudates from soybean seeds at three seed quality levels for two cultivars and to determine the Pythium disease development by four Pythium spp. in three seed quality levels for two cultivars. Experiments consisted of two soybean cultivars (Archer: flood tolerant and Hutcheson: a widely grown cultivar), three quality levels (high: above 80, medium: 60-79, and low: below 60 percent germination) and five Pythium treatments: P. aphenidermatum, P. vexans, P. irregulare, P. ultimum and none. These treatment combinations were tested at 20°C in a factorial arrangement. Seed exudates collected from the three seed quality levels of the two cultivars at 20°C were analyzed for organic acids and sugars. Seed germination/seedling emergence were evaluated and root discoloration were rated on 0-7 scale. The study concluded that the exudation of sugars and organic acids was less in Archer compared to Hutcheson and less in high quality seeds and increased with decreasing seed quality in both cultivars. Also, nutrient exudation was highly correlated to Pythium disease development.

36 Budapest, Hungary, May 17th – 19th, 2004

17 - APPLICATION OF ADVANCED TECHNOLOGIES IN MAIZE (ZEA MAYS L.) PÁL, PEPÓ

University of Debrecen, Centre of Agricultural Sciences, Faculty of Agricultural Sciences, Department of Genetics and Plant Breeding, Debrecen

Currently, the most important crop of the Hungarian production is maize. Therefore, it is very important to concern with the problems of production, and use of maize and at last but not at least the difficulties in breeding and the future tasks of maize production. The proper choice of hybrids predetermines the success of maize production, the yield and the stocks for sale which gives the results for farmers. Mutation breeding is a great help for widen the genotypes suitable in maize selection. Application of induced mutants increases the gene reserves which has a great importance because of the reduction in variation of populations. With the help of mutation we increased the variability and selected lines favorable for plant production. We produced new hybrid combinations to satisfy the production requirements. We completed diversification of the basic breeding material with neutron radiation. Use of neutron sources in maize breeding programs in a larger scale is well based because of its great genetic affinity. After the seed treatment with fast neutron source we self-pollinated the segregated population, created genetically homozygote lines and selected the stable inbred lines with favorable agronomic characters. We have concluded that the cyclotron can be successfully applied in widening the genetic variability. We have produced a number of inbred lines with wide genetic variability using mutation breeding: with application of diallele analysis we have selected the lines with the best general and specific combining ability. Using these lines we can create F1 hybrids which have the most valuable economic features. In order to complete these tasks we have to examine diallele experiments with many replications in a number of vegetation periods and investigate many factors. After summarising results we can get conclusions to further selections. We concluded that only those lines and their hybrids can form the base of further breeding programs which give prominent results and fulfill the requirements. It is an important task to increase the productivity of the maize and improve the economical characters of the crops. Our results suggest that we have to take into account not only the yield and its elements, but their physiological processes. At the end we have to select on the optimum of the factors mentioned above.

18 - RELEVANCE OF THE GENETIC BACKGROUND OF THE SEED TO THE APPLICATION OF REAL TIME PCR STRATEGY FOR GMO QUANTIFICATION IN MAIZE SEED SAMPLES PAPAZOVA1,2, N., DEGRIECK2, E., VAN BOCKSTAELE2,3, E., JOOS4, H. & DE LOOSE2 M.

1AgroBioInstitute, blvd. “Dragan Tzankov” 8, Sofia 1000, Bulgaria, [email protected] 2Department of Plant Genetics and Breeding, Center for Agricultural Research, Caritasstraat 21, 9090 Melle, Belgium, [email protected] 3Department for Plant Production, Ghent University, Faculty of Agricultural and Applied Biological Sciences, Coupure Links 653, 9000 Gent, Belgium 4Bayer BioScience N.V., Josef Plateaustraat 22, 9000 Gent, Belgium

Real Time PCR nowadays is a method of choice for quantitative determination of GMO impurities in plants and plant products. The output of this analysis is number of haploid transgenic genomes related to the total number of genomes which has to be converted to number of transgenic seeds to the total number of seeds as it is accepted to express the GMO impurities in seed samples. The existence of the structures with different genetic background (diploid embryo and triploid endosperm), that contain different DNA quantity in the maize seed is a factor that can influence quantitative Real Time PCR based analysis. The influence of the genetic background was estimated using two approaches:

37 27th ISTA Congress – Seed Symposium

• Theoretical – non-transgenic variety was used for determination of the ratio between the endosperm and embryo DNA in the maize seed. Based on the obtained data the minimal requirements for heterozygous seed to be GMO are calculated. The use of lower working threshold for Real Time PCR for maize corresponding to the threshold for seeds (0.5%) is proposed. • Practical – determination of the GMO content in heterozygous seed of transgenic maize line T25. Seeds with different inheritance of the transgene were obtained after reciprocal crossing of T25 with non-transgenic variety. The data are in compliance with the model defined by the theoretical approach.

19 - ENCAPSULATION OF ANDROGENIC EMBRYOS AND PVRO-EMBRYOS FOR PRODUCTION OF SYNTHETIC SEEDS IN ELITE INDICA RICE VAR. IR72 ROY, BIDHAN & MANDAL, ASIT B.

Biotechnology Section, Central Agricultural Research Institute, Port Blair 744 101, India, [email protected], [email protected]

Embryos and pro-embryos were produced in large quantity from androgenic callus developed on MS medium for production of synthetic seeds in indica rice (Oryza sativa L.) var. IR72. High concentration of BAP (4-6 mg L-1) was found to yield a large number of dormant embryos and pro-embryos in about one and half months time. Thus these embryos and pro- embryos are the rapid and recurrent source for production of synthetic seeds. Those individual embryos, pro-embryos and embryo like structures were encapsulated in sodium alginate (2.5% w/v). In vitro germination and plantlet regeneration of the encapsulated seeds were assessed by germinating them on MS containing different combinations and concentrations of cytokinin (BAP, Kinetin) and auxin (NAA). High germination (87.5-55.0%) was recorded when MS was supplemented with BAP and lower concentration of NAA. To contrary, addition of Kinetin in MS drastically reduced the germination percentage. The germination of unbeaded pro-embryos was 92.5% on MS basal medium. Reduced germination of artificial seeds might be due to the damage incurred while separating the embryos from cluster and/or owing to adverse effect of chemicals towards germinability used for encapsulation. Moderate germination (40.0%) was observed on sterile sand in in vivo. Such synthetic seeds may be used for in vitro propagation as well as genetic transformation experiments with ease as an alternative of initiation of fresh tissue culture for induction of callus and propagules for genetic modulation experiments.

20 - COMBINING ABILITY AND HETEROTIC STUDIES OF ELITE INBREDS WITH DIVERSE CMS SOURCES IN SUNFLOWER (HELIANTHUS ANNUUS L.)* GOURI SHANKAR1, V., GANESH2, M. & RANGANATHA3, A.R.G.

1M.Sc (Ag) student, ANGRAU. India 2Principal Scientist, National Seed Project, Hyderabad, India 3Senior Scientist, Directorate of Oilseeds Research, Hyderabad, A.P., India.

Majority of the presently grown commercial sunflower hybrids in the world possess cytoplasm of Helianthus petiolaris (PET 1). Dependency on a single source of cytoplasm, may lead to genetic vulnerability of hybrids, if the cytoplasm becomes susceptible to a new strain of disease or insect pest. It is evident from the earlier studies that the dependency on single source of cytoplasm posed a threat to the cultivation of maize and bajra, as they became susceptible to the leaf blight and downy mildew, respectively. As the area under sunflower substantially increased that too depending on single source of cytoplasm i.e. PET 1, is posing problem for most of the abiotic and biotic stresses. In this direction, sunflower breeders of Directorate of Oilseeds Research, Hyderabad, accelerated their effort to diversify CMS sources and to identify effective restorers. In the present study, eight diverse CMS lines (representing three sources viz. ‘PET 1’, ‘CMS I’ and ‘CMS PEF’) were crossed with 18 elite inbreds (developed through population improvement programme). The resulting 144 hybrids were studied for Maintainer/Restorer reaction. Of these, 103 hybrids showed fertility and rest were behaved as sterile. The inbreds, which behaved as maintainers can be converted into

38 Budapest, Hungary, May 17th – 19th, 2004 new CMS lines in future breeding programmes. Based on the fertility restoration, a set of 77 F1s (7 CMS lines x 11 inbreds) were selected which fits into Line x Tester design to know the combining ability. It was observed that the lines ARM 243B and ARM 238B and the testers R- 856 and R-297 were good general combiners while PF-400A x P-356, PF-400A x R-273 and IMS-852A x R-265 cross combinations with diverse CMS sources were showed significant sca effects for yield and most of the yield components. Preponderance of non-additive gene action was noticed for all the characters. The crosses IMS- 852A x R-17 (366.50), ARM 245A x R-17 (295.52) and 302A x RHA-6DIR (263.24) per cent of heterosis was noticed for seed yield. These crosses can be further evaluated for G x E interaction before they release as hybrids. Besides ‘PET 1’, other sources viz., ‘CMS I’ and ‘CMS PEF’ sources used in the present investigation were equally efficient in expressing their fullest potential of yield and yield contributing characters. Thus breeder can safely use these sources to broaden the genetic base, thereby sunflower can be safeguarded from any eventuality due to biotic and abiotic threats in future. * It is a part of M.Sc. (Ag) thesis work on sunflower.

21 - DIFFUSION TECHNOLOGY AS A TOOL FOR CHARACTERIZING AND ENHANCING GERMINATION OF MAIZE (ZEA MAYS. L) GENOTYPES SOOD1, M.L., ZELEKE2, H. & MESGANAW, M.

Department of Chemistry1 and Plant Science2, P.O. Box 298, Alemaya University, Ethiopia.

Diffusion technology employed earlier for characterizing seed genotypes and the theory of water diffusivity recently developed has now been further extended and tested for enhancing germination of two promising maize (Zea mays.L) genotypes viz. BH-660 and Pop-902x903. The values of the seed constants obtained clearly showed that the two maize genotypes are genetically different not only from one another but also from the four Indian maize genotypes viz. Paras, ParKash, Parbhat and Kesri. The water diffusivities at 25oC were found to be 0.0651x10-5 cm2 s-1 and 0.0556x10-5 cm2 s-1- respectively indicating that the diffusion rate of water in BH-660 is 1.2 times greater than Pop- 902x903 and it absorbs nearly 10.20% more water than the latter genotype. Field trials results have shown an increase of 8% and 2% in seed germination when the seeds of the two genotypes were sown after these were subjected to water diffusion at 25oC and steady- state conditions obtained in the laboratory. These studies have clearly demonstrated that the critical moisture content and physiological level attained at the steady-state conditions during water diffusion bears a direct relation with seed vigor and germination and the technology so developed can be fruitfully exploited with seeds of other maize genotypes and crops.

22 - DEVELOPMENT OF DNA PROTOCOLS FOR NATIVE TREE SPECIES FROM THE BRAZILIAN ATLANTIC FOREST SOUSA, V.A., SANTOS, E. C., WENDT, S. N., MEDEIROS, A. C. S. & SANTOS, A. F.

Embrapa Florestas – National Forest Research Center of EMBRAPA, Cx. P. 319, 83411-000, Colombo, PR – Brazil, [email protected]

The area of Brazilian Atlantic Forest has been drastically reduced as result of indiscriminate exploitation. Species such as “aroeira” (Schinus terebinthifolius - Anacardiaceae), “maricá” (Mimosa bimucronata - Mimosaceae) and “pata-de-vaca” (Bauhinia forficata - Caesalpinaceae) are threatened of extinction. Therefore measures of conservation are urgently need involving the remaining populations. Embrapa Florestas established in Colombo, State of Paraná, Brazil, a native tree seed bank, named BASEMFLOR, in order to provide seeds to be used in forest restorations, genetic conservation and research programs. Basic studies on pattern of genetic variation between and within populations to guide the seed collecting are necessary. Also, to support different programs in maintaining a high genetic diversity that represent the success of most forest programs. Actually many molecular techniques are applied to detect the genetic variability at DNA level. The aim of this work was to develop extraction DNA protocols to support genetic analysis of different species of Brazilian Atlantic Forest. The DNA extraction protocols for “aroeira”, ”maricá” and 39 27th ISTA Congress – Seed Symposium

“pata- de-vaca” were successfully established. The screening of the primers are now been carried out and the genetic analysis of populations with RAPD technique will be conducted soon.

23 - TESTING GENETICALLY MODIFIED SEEDS OF AGRICULTURAL PLANTS IN SLOVENIA ŠUŠTAR-VOZLIČ, J. & MEGLIČ, V.

Agricultural Institute of Slovenia, Hacquetova 17, Ljubljana, Slovenia, [email protected]

Recent advances in plant biotechnology have produced many new genetically modified (GM) crop varieties with advantageous characteristics, which are now being grown in many countries worldwide. In some European countries, the cultivation and marketing of GM plants is done together with the conventional non-GM varieties. The adventitious presence of GM seeds in non GM seed lots has increasingly become a problem for the international seed trade. Requirements for the quality of seed material and seed trading are prescribed and regulated through international laws which are already in force in Europe or are being in preparation (e.g. the new Directive and Regulation for seeds). In Slovenia this field is since 2002 regulated by a National Law on management of genetically modified organisms and the Law on agricultural seeds and propagating material. At the moment there are no GM varieties of any agricultural crop species allowed to be grown in Slovenia and there are also no field trials established. The Seed Testing Laboratory at Agricultural Institute of Slovenia (AIS) is the only ISTA accredited laboratory in Slovenia, which is appointed by the Ministry of Agriculture, Food and Forestry for the analysis of imported and exported seeds. To follow the international and Slovenian legislation and to enable a continuous domestic and international seed trade, the Seed testing laboratory at the AIS has started with establishing the protocol for testing the presence of GM seeds in conventional seed lots of maize and soybean. Among various analytical methods, that have been developed to reliably determine the presence of genetically modified seeds of agricultural plants, qualitative PCR based methods were applied. Detection of genetic elements commonly used in the GMO constructs (e.g. 35 S promoter or nos terminator) is used as a screening method. The protocols for detection of all four in Europe permitted maize lines (MON810, Bt-11, Bt-176, T25) and soybean line Roundup Ready were established using event specific primers. The quantification of the GMO level in positive samples is done by means of semi-quantitative tests, therefore various sub- sampling schemes are being tested.

24 - AFLP ANALYSIS OF GENETIC DIVERSITY AMONG ITALIAN POPULATIONS OF TRITICUM DICOCCUM TALAME’, V., BALLARDINI, M., NOLI, E., D’ANTUONO, F. & TUBEROS, A R.

Department of Agroenvironmental Science and Technology, ‘Alma Mater Studiorum’ Università di Bologna, Viale G. Fanin 40, I-40127 Bologna, Italia

Triticum dicoccum (emmer wheat) is one of the three domesticated species of hulled-wheat (T. monococcum L., T. dicoccum Schubler and T. spelta L.) which is still traditionally grown in limited and marginal areas of central Italy in poor fertility soils. This species has recently been the object of agronomical and genetic studies due to the growing interest towards the typical dishes of the regional cuisine. The cultivars grown in Italy are genetically heterogeneous landraces. In this study, seven plants sampled from each of ten Italian populations of emmer (six winter- and four spring-type), collected from their traditional cultivation areas, were analysed by amplified fragment length polymorphisms (AFLPs) markers in order to characterize their level of genetic variability. Eight AFLP primer combinations were used for selective amplification yielding 146 polymorphic AFLPs. On an average, each assay revealed 19.1% polymorphic bands. For each AFLP assay, the Index of Polymorphic Content and the Marker Index indicated a good discrimination among plants.

40 Budapest, Hungary, May 17th – 19th, 2004

The average genetic similarity (GS) among the 70 plants, estimated according to the Dice coefficient, was 0.64, with higher values among plants with spring- than with winter-habit (GS = 0.83 and 0.79, respectively). Cluster and principal component analysis showed a clear separation of the plants among populations and between growth habits. The correlation values between similarity estimates computed for each AFLP assay and for the whole set of primers allowed for the identification of a number of highly informative AFLP assays that will be useful to further characterise T. dicoccum populations.

25 - DIGITAL IMAGES ANALYSIS IN CORN SEEDLING EVALUATION TEIXEIRA EVERTON, FELIX, CICERO, SILVIO MOURE & DOURADO, NETO DURVAL

São Paulo University, Av. Pádua Dias, 11, C.P. 09, 13418-900 Piracicaba – SP, Brazil, [email protected]

The image analysis has high potential use in seed vigor determination. Associated to the seedlings growing test, this technique is fast, precise and makes possible the dimensional image analysis. The result is the total extension of each seedling quantifying the length of their constituent parts. The objective of the present paper was to study the vigor of corn seed lots through the seedlings growing test, using the digital images analysis. Corn seedlings by AG 122 cultivar were retired from the germination chamber at fourth day of development and ordered over a sheet made with a transparent polyester film on a scanner surface to the images caption. A routine was developed to process digital images of seedlings into the “Scil-Image” software. There was a computational procedure to measure the total length, with the usual sum of coleoptiles to the length of the larger seedling root and also, not usually, to all root system. The seedlings were measured manually, seeking comparison with the method in study. The results showed that the digital technique makes possible association of the data obtained in processing to eventual vigor differences existing in corn seed lots, in a similar way to the other methods appointed to seed vigor evaluation by referred species.

26 - DEVELOPMENT OF MICROSATELLITE MARKERS FOR THE IDENTIFICATION OF BRAZILIAN COFFEA ARABICA CULTIVARS. VIEIRA1, ELISA S.N, VON PINHO1, ÉDILA V.R., ESSELINK2, DANNY G.; VIEIRA, MARIA G.G.C.1; VOSMAN, BEN2

1Federal University of Lavras, Lavras, Minas Gerais, Brazil, [email protected] 2Plant Research International, Wageningen, The Netherlands

Microsatellite markers or SSR (Simple Sequence Repeats) have proved to be an excellent tool for cultivar identification and genetic relationship studies in plants. A set of 140 SSR markers were used to analyze the genetic similarity among twenty five Coffea arabica cultivars composed of nineteen brazilian cultivars of commercial importance, and six indian hybrids of Coffea arabica, Coffea canephora and Coffea liberica. Of the total number of SSR markers tested 127 DNA nuclear markers were developed using enriched small insert libraries and the coffee SSR sequences available in the NCBI database, and thirteen universal chloroplast DNA markers were also tested. Among the twenty-two polimorphic loci 2-7 alleles were detected for each locus with an average of 3.5 alleles per locus. The locus CAga001 was the most discriminating for brazilian cultivars with 4 allelic phenotypes and 1.9 effective alleles. The mayority of the SSR loci contained di-nucleoditde (GT) repeats and the polymorphism was positively correlated with the number of repeats. Based on the band patterns generated by the polymorphic SSR loci, the set of twenty-five coffee cultivars were clustered in two main groups. One group composed of the most part of the brazilian cultivars and a second one composed of the indian hybrids. Many of the color mutants were not separated. The clustering was in accordance with the background of the cultivars and showed the high level of similarity of the brazilian material, indicating the necessity of new introductions or crosses between cultivars with different background in future breeding programms.

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27 - VARIETAL IDENTIFICATION IN BRINJAL (SOLANUM MELANOGEN L.), PALAK (SPINACIA OLERACEA L.) & PEA (PISUM SATIVUM L.) BY GEL ELECTROPHORESIS OF SOLUBLE SEED PROTEINS VIG, ADARSH PAL & KAUR, RABINDER

Guru Nanak Dev University Amritsar, Department of Botanical and Environmental Sciences, Amritsar – 143005, India, ([email protected])

Varietal identification is one of the most important aspect of seed industry and seed trade. With the increase in number of varieties in each crop it has become difficult to identify and characterize these varieties on the basis of morphological characters alone. Thus it has led to the exploration of new stable characters including genetic make-up to be used as markers for varietal identification. The present investigation includes identification of 4 varieties of Brinjal, 2 varieties of Palak and 7 varieties of Pea. These varieties were analyzed on the basis of their protein profile. Proteins were extracted from dry seed samples of different varieties by using sodium phosphate buffer. The extracted protein samples were run at 120-200 volts for 5 hours using Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) at 12% concentration. After staining, banding pattern was obtained. Data was represented in the form of electrophoregrams and analysed on the basis of presence or absence of bands and staining intensities. In case of Brinjal the variety BH2 Hybrid was showing a clear distinction from other varieties by the presence of band at position 4 which was not present in any other variety. Moreover presence of two dark bands in this variety also differentiate this variety from the rest of the three varieties which had only one dark band each. In Sada Bahar presence of two faint bands made this variety different from others as no variety was having bands of faint intensity. In case of Palak it was observed that both the varieties were similar in their banding pattern and intensity. In case of Pea, presence of all the 11 bands in Bonneville differentiate this variety from rest of the varieties as in no other variety all the 11 bands were present. Presence of band at position 4 in Mithiphali and at position 3 in SM-1 differentiate these varieties from other varieties as no band at these positions were observed in any variety except Bonneville. Presence of light band at position 2 in SM-1 variety differentiate this variety from other as no light band was present in any other variety at this position. Results of this study clearly showed that varieties could be easily distinguished using SDS – PAGE .

28 - GENETIC PURITY TESTING OF TWO-LINE HYBRID RICE SEEDS BY GROW-OUT OF EMBRYO-CONTAINING HALF-SEEDS AND ULTRATHIN-LAYER ISOELECTRIC FOCUSING OF PROTEINS FROM CORRESPONDING EMBRYOLESS HALF-SEEDS WANG, XIAOFENG, ZHAO, TING, YAN, MIN & LU, YANPENG

South China Agricultural University, China, [email protected]

To test for inbreds among F1 seeds of two-line hybrid rice (Oryza sativa L.) in the laboratory, ultrathin-layer isoelectric focusing (UTLIEF) of seed proteins was used. One female marker band in the photo-thermo-sensitive genic male sterile rice (cv. Peiai 64S) and one common male marker band in the four restorer lines (cv. G67, Peifu, I9-1 and Minkezhan) were found in the pH range of 5-8. Embryoless half-seeds contained the same male marker proteins as whole seeds and could be used for UTLIEF to test inbreds in the laboratory. The corresponding embryo-containing half-seeds could germinate to form seedlings on vermiculite and seedlings be transplanted into the field for grow-out tests. The genetic purity determined by UTLIEF of proteins from embryoless half-seeds was well and truly confirmed by grow-out tests of the corresponding embryo-containing half-seeds. This work shows that UTLIEF of seed proteins could replace grow-out tests to determine F1 genetic purity of two- line hybrid rice. Using half-seeds, the comparison between laboratory and field tests was more objective and accurate than using whole seeds.

42 Budapest, Hungary, May 17th – 19th, 2004

29 - A STUDY ON MATCHING AND MARKING TECHNIQUES OF SEED DYNAMICAL IMAGES BASED ON MACHINE VISION LI WIE1, LIN JIA-CHUN2, ZHAN BIN2 & TAN YU-ZHI2

1College of Engineering China Agricultural University Beijing 100083, China 2Department of Machine Engineering, China Agriculture University, Beijing 100083, China [email protected], [email protected]

This paper discusses a new multi-seeder dynamic testing technology that makes it possible to accurately and reliably measure seed space intervals using image processing and analysis techniques. A dynamic test of a precision seeder at step-less speed (0.5m/s-3m/s) was developed based on machine vision. We also discuss the development of a universal test-bed designed to test the performance of precision seeders and grain drills along with the simultaneous capture of seed distributions from the three seeding monomers. In this paper, according to the demand of real time system, segmentation technique of seed images based on automatic threshold selection is studied. The images are segmented by using a combination of static threshold segmentation and adaptive threshold selection based on iterative algorithm. The seeds are recognized by labeling different areas with different labels according to the pixels grey level comparability in the area, and at the same time, the system marks and seeds are recognized and positional parameters are calculated. Finally, many methods was made based on a pattern recognition technique. By analyzing influence diathesis of image joining precision, image joining algorithm and setting of overlapped area are proposed. A universal test-bed system is expounded that can be used to test the performance of precision seeders, grain drills and seeding monomer. This test bed can measure the seeds’ interval correctly and reliably and operate automatically. Experiments show that theory and method are practical, speedy and reliable, the demanding of real time is satisfied, with an error in seed interval of ±2 mm.

30 - CHARACTERIZATION OF RAPESEED, MUSTARD AND TARAMIRA CULTIVARS USING MATURE PLANT CHARACTERS YADAV1, SHIV KUMAR, YADAV2, T.P. & RAM3, C.

1Division of seed science and Technology, IARI, New Delhi-110012, India 2Oil Seed Section, Department of Plant Breeding, CCS Haryana Agricultural University, Hisar – 125004, India 3Department of Seed Science and Technology, CCS Haryana Agricultural University, Hisar – 125004, India

Eighteen mustard (Raya) i.e. Brassica juncea (L), four rapeseed (toria) i.e. Brassica campestris (L). and one cultivar each of Brassica campestris var. Yellow Sarson, Brassica campestris var. Brown Sarson, Brassica napus (L). and Eruca sativa (L). (Taramira) were studied for 14 various quantitative and qualitative characters for two years. Mature plant characters like growth habit, mature leaf shape, mature leaf color, intensity of mature leaf color, stem pubescence, mature plant height, number of primary branches, number of secondary branches, main shoot length, morphotype uniformity and basal side shoot development were used to classify the varieties, which provided a sound classification system as these characters exhibit distinct varietal patterns. The mature leaf color, morphotype uniformity and plant height were the plant characters that showed continuous differences during both the seasons. Hence, all the other characters studied were found useful for the purpose of cultivar identification. A list of key characters useful in variety identification was also prepared. It was possible to distinguish all the cultivars individually using the seed key.

43 27th ISTA Congress – Seed Symposium

31 - ESTABLISHMENT OF LOW-COST AND HIGH-THROUGHPUT SSR ANALYSIS CONDITIONS FOR VARIETAL IDENTIFICATION IN MAIZE. ZHANG, D. BAZENTE, F. & LALLEMAND, J.

GEVES, Laboratoire BioGEVES. Le Magneraud. BP52. 17700 SURGERES, France, [email protected]

Our presentation will describe an efficient SSR analysis system using a LI-COR automated DNA analysis system. This system consists of (1) multiplexing PCR, (2) tailed one of the two primers for each SSR primer pair, (3) pooling two independent PCR runs each using a different dye for tail labelling before gel loading, and (4) reloading at least three times a same gel. Secondly, a set of 50 SSR markers, well distributed on the maize genome, was established for routine genotyping study in maize, after their characterization on a set of 45 public maize inbred lines. The cost of routine genotyping using our SSR analysis condition was estimated from sample preparation, DNA extraction, PCR and to data analysis. It varied according to the number of SSR markers to be used, from 1 euro per data point if only 6 SSR markers are used to 0.5 euro per data point if 30 SSR markers are used.

44 Budapest, Hungary, May 17th – 19th, 2004

2. Organic and Conventional Seed Production Topics: Seed crop management; harvest; processing; post-harvest technology; seed certification

32 - THE EFFECTS OF ENVIRONMENTAL CONDITIONS DURING FILLING OF BOLL ON SUBSEQUENT GERMINATION AND SEEDLING GROWTH OF THE COTTON AKRAM-GHADERI1, FARSHID, SOLTANI2, AFSHIN & REZAEI1, JAVAD

1Cotton Research Institute, Gorgan, Iran, ([email protected]) 2Department of Agronomy, Gorgan University of Agricultural Sciences, Gorgan, Iran

Three cotton cultivars of Sahel, Siokra324 and Deltapine25 were sowed in different dates, and their seeds were harvested at the stage of the opening of the boll. Then germination test was conducted on these seeds. The percent of germination, the length of root and shoot and the dry weight of the root and shoot and the total seedling dry weight were measured. Results showed that the planting date did not have any significant effect on the germination percent and the length of the root and shoot. But with delay in planting date, the dry weight of root and shoot and the total seedling dry weight in Deltapine25 decreased significantly. However, two other cultivars were not affected by planting date. These results show that under the condition of this experiment, the seeds of Deltapine25 cultivar that were produced from the earlier planting dates have more seedling growth rate compared with produced seeds from the later planting dates. But the seedling growth rate was similar for other two cultivars obtained from different planting dates.

33 - NEW METHODS TO DETECT SOYBEAN SEEDS TOLERANT TO GLYPHOSATE TILLMANN¹, MARIA ANGELA ANDRE, MEDIANEIRA BERTAGNOLLI2, CARLA, MECCA DE MENEZES2, SABRINA, BICCA DODE3, LUCIANA & AMARAL VILLELA¹, FRANCISCO

¹bolsista CNPq,UFPel/FAEM, Caixa Postal 354, CEP 96001-970, Pelotas-RS, [email protected] 2UFPel/FAEM, Pelotas-RS; 3UCPEL, Pelotas-RS)

With the objective of differentiating soybean cultivars, with respect to resistance to the herbicide glyphosate, two assays were carried out using a parental soybean (non-GMO) and the Roundup Ready® (RR) genotype (GMO, resistant to glyphosate). In the first assay, peroxidase electrophoretic patterns were analyzed. The seeds were pre-imbibed in water or in glyphosate solution containing 0.6% active ingredient (N-phosphonomethyl glycine, 360 gL-¹ formulated product) 16 h. and incubated for 7 days at 25ºC in the germinator. Peroxidase activity of extracts was determined by colorimetric reaction and by electrophoretic patterns. In the second assay, a protocol was established to detect RR seeds in hydroponic cultures. Seeds from the two genotypes were laid out on a specially designed mesh located on the lids of trays containing glyphosate at different concentrations (0.0, 0.12, 0.24, 0.36 and 0.48%). The trays were placed in illuminated tanks containing water at 25ºC for 4 hours. The trays were transferred to tanks containing a nutrient solution and aerated for 6 days. The seedlings were evaluated for root length, the hypocotil and the total number of seedlings and secondary roots. A differential peroxidase activity, assayed by electrophoretic patterns, between the resistant (RR) cultivars and the parental cultivars was observed. The hydroponics assay allows the detection of seeds resistant to glyphosate (0.12% glyphosate, 4 hours assayed on day 6). Both assays were found to be efficient in the identification of the RR soybean genotype.

45 27th ISTA Congress – Seed Symposium

34 - EFFECT OF AGE OF SEEDLINGS ON FLOWERING BEHAVIOUR OF PARENTAL LINES OF RICE HYBRID BIRADARPATIL, N.K. & SHEKHARGOUD, A.M.

University of Agricultural Sciences, Dharwad-580 005, India

Transplanting of rice seedlings at the right age assumes special significance in obtaining higher yields in commercial cultivation as well as in hybrid seed production. Earlier reports indicate that the age of the seedlings influences the flowering, and this information is useful in hybrid seed production for achieving the flowering synchrony. Hence, a study was carried out to know the influence of age of seedling on flowering behaviour of parental lines of KRH- 2 rice hybrid with seedlings of different ages (15, 20, 25, 30, 35, 40, 45 and 50 days). The results indicated that the increase in the age of the seedlings increased the days to flowering of CMS, maintainer and restorer lines. On an average, there was about eleven day’s difference in days to 50 percent flowering between 15 days and 50 days old seedling. The CMS line (IR 58025A), the maintainer line (IR 58025B) and the restorer line (KMR-3R) took 96.7, 91.3, 104.3 days, respectively, for 50 per cent flowering with 15 days old seedling, whereas, with 50 days old seedlings they took 107.7, 103.7 and 117.3 days, respectively. The difference in flowering between the restorer and male sterile line increased with the increase in the age of the seedlings.

35 - QUALITY OF HARD WHEAT SEEDS AFTER DRYING AND STORAGE PROCESSES COLPETTI, ANDRÉ, BIAG, I. JOÃO & CARNEIRO, LUCIANA

1Campinas State University – FEAGRI, Campinas, Brazil, [email protected], 2FEAGRI/UNICAMP, [email protected] 3FEAGRI/UNICAMP, [email protected]

A wide range of products can be obtained by wheat grains and seeds. There are some hard wheat that do not present dormancy; they may germinate on ear between the maturation and harvest processes if pluvial precipitations occur. The objectives of this work were to evaluate the effects of moisture content and storage on the quality of hard wheat seeds. The experimental work was installed at Campinas Agronomy Institute. The seeds of IAC-1003 (Triticum durum L.) were harvested with three different moisture contents: 19.9%, 16.9% and 10.2%. The seeds of 1st and 2nd harvests were dried with heated air at 40, 60 and 80ºC. After the drying process, the seeds were stored in polyethylene packets for a period of 0, 2, 4 and 6 months at 20±2ºC. During the storage, tests of quality, water activity, protein and carbohydrate percentages were carried out in the Post-harvest Technology Laboratory, FEAGRI/UNICAMP. The drying temperature of 80ºC reduced the germination and vigour of the seeds, however the drying temperatures of 40 and 60ºC did not affect the seeds. The results of the protein and carbohydrates percentages of IAC-1003 seeds did not show significant difference during the storage period.

36 – EARLY HARVEST DRYING AND STORAGE AND THE QUALITY OF SEEDS OF COMMON AND HARD WHEAT CARNEIRO, LUCIANA1, BIAG, I. JOÃO1

1Campinas State University – FEAGRI, Campinas, Brazil, [email protected], [email protected]

Seeds of wheat that do not present dormancy may germinate in the spike, making impracticable the use of the seeds. The objective of the work was to verify the influence of time of harvest, drying and storage in the quality of wheat seeds. The seeds used in this study were from genotypes of common wheat (Triticum aestivum L.), one with dormancy (IAC-24), harvested with 30, 21.2 and 12.2% of moisture content and one without dormancy

46 Budapest, Hungary, May 17th – 19th, 2004

(IAC-289), harvested with 35, 23.4 and 12.5% of moisture content; 1 genotype of hard wheat (Triticum durum L.) without dormancy (IAC-1003), harvested with 31.6, 22.2 and 11.7% of moisture content. The seeds of 1st and 2nd harvest were dried at 40, 60 and 80ºC until they reached 13.0% of moisture content. Dried seeds were then stored in polyethylene packets for a period of 0, 2, 4, 6 and 8 months at 20±2ºC, and were evaluated using the moisture content tests, 1000-seed weight, germination and vigour. The drying to 80ºC, reduced the germination and the vigour of the seeds, mainly with the storage. The early harvest did not affect the quality of seeds of the IAC 24 and IAC 289 cultivars, however for the IAC 1003 seeds, harvested with 31.6% moisture content the germination and vigour were affected.

37 - AN X-RAY EQUIPMENT FOR SEED QUALITY ANALYSIS CRAVIOTTO1, R.M., ARANGO1, M.R., SALINAS2, A.R., GIBBONS3, R., BERGMANN3, R. & MONTERO. M.S.

1E.E.A. INTA Oliveros, Ruta Nacional Nº11 Km 353 (2206) Oliveros; Santa Fe, Argentina, [email protected] 2Facultad de Ciencias Agrarias (Faculty of Agricultural Sciences) - UNR (National University of Rosario, [email protected]) 3Industrias Texel.

The well-known X-ray technique is commonly used in Medicine, Biology and a variety of industries. The International Rules for Seed Testing issued by ISTA include a chapter devoted to this technique application, and the Association of Official Seed Analysts (AOSA) published an X-ray handbook on the use of this technique for different species. The main objective of this study was to develop a prototype of an X-ray equipment with applications specific to seed quality control. The equipment consists of: A cabinet made of a casing and a metallic structure as support, a sample holder structure, a power source and electric circuit, an X-ray transmitter (tube), a computer, a Visualix system, which consists of: X-ray sensor (CCD), auxiliary electronic interface, electronic card (Frame Grabber), and computer imaging software (VixWin). With this equipment, images can be quickly obtained, maintained, reproduced and scanned at any time. The equipment developed, called X-ray Seed Analyzer (ARX), uses a non-destructive method, which complies with the security standards required by Sanitary Radiophysics. It is a useful tool for the analysis of seminal structures, for the differentiation of seeds and/or grains full, useless, physically damaged, or with eggs or living insects inside, allowing a frequent and rapid monitoring with a high level of reliability in the diagnosis.

38 - ACTIVE SEED BANK OF NATIVE TREE SPECIES FROM THE BRAZILIAN ATLANIC FOREST - BASEMFLOR ®. DE S. MEDEIROS, A. C., DOS SANTOS, A. F. & DE SOUSA, V.

Embrapa Florestas – National Forest Research Center of EMBRAPA, Cx. P. 319, 83411-000, Colombo, PR – Brazil, [email protected] The present status of the Brazilian Atlantic Forest is the result of nearly five centuries of exploitation since the colonial times and land use for the expansion of agriculture as well as for the development of urban areas. It is estimated that the remnants cover only 4% of the original forest area. According to the Brazilian Report for the United Nations Conference on the Environment and Development, this forest stretches through nearly one million square kilometers in almost all coastal areas of the country. Effective actions for the restoration of forest cover have been extremely shy. The main bottleneck has been the inexistence of an active seed bank for the species of native forests. It is of utmost importance that such a seed bank be part of a federal level research institution in order to provide seeds for uses such as reforestation programs, reclamation of degraded ecosystems, establishment and restoration of riparian forests and forest covers on water catchment areas, botanical gardens, local and regional seed banks, and for research projects. As a contribution to solve part of the problem, Embrapa Florestas set up an active gene bank in the form of seeds, established in Colombo, State of Paraná. 16 species from 11 different families (Anacadiaceae, 47 27th ISTA Congress – Seed Symposium

Aquifoliaceae, Leguminosae-Fabaceae, Melastomataceae, Meliaceae, Mimosaceae, Myrsinaceae, Sapindaceae, Saxifragaceae, Tiliaceae and Winterceae) were collected and stored (at 5°C) in 2002-2003 for distribution. This BASEMFLOR® involves research on seed physiology, seed pathology, entomology, genetic characterization, training, and the establishment of a data bank to convey all information through the computer network.

39 - THE INFLUENCE OF STAGE OF FRUIT MATURATION ON SEED QUALITY IN TOMATO (LYCOPERSICON ESCULENTUM MILL.). DIAS; D.C.F.S., RIBEIRO; F.P., DIAS; L.A.S. & SILVA, D.J.H.

Department of Fitotecnia, Universidade Federal de Viçosa – 36571-000, Viçosa, MG, Brazil, [email protected]

The effects of stage of fruit maturation at harvest and post-harvest period on seed quality of tomato were investigated. Fruits of Santa Clara cultivar were harvested in three different maturation stages: S1 – fruits with the outside still green, but with initial red colour on the top; S2 – fruits completely red (fully ripe); and S3 – fruits harvested at S1 and kept on storage under packinghouse conditions until S2 stage. The physiological quality of seeds was evaluated by standard germination (normal seedlings after 14 days), germination at first count (normal seedlings after 7 days), controlled deterioration (seeds with 24% moisture content in water bath at 45°C for 24 h), electrical conductivity (four replications of 50 seeds imbibed in 50 mL distilled water at 25°C for 24 h) and speed of seedling emergence. The germination percentage of the seeds obtained from fruits harvested on S3, S1 and S2 stages was 88, 83 and 82%, respectively. For the same treatments were obtained 72, 47 and 51% of normal seedlings at first count and 78, 57 e 57% on controlled deterioration test, respectively. For all characteristics evaluated, post-harvest storage of the fruits improved seed vigour. Therefore, tomato seeds should be extracted from fruits harvested at S1 stage and submitted to storage until completely ripe.

40 - METHODS FOR ORGANIC SEED TREATMENT ERTSEY, A. & RADICS, L.

1BUEPA, Faculty of Horticultural Sciences, Department of Ecological and Sustainable Production Systems, H-1118, Budapest, Villányi út 29-43, [email protected], [email protected]

The importance of ecological farming in Hungary is increasing. In the last twelve years size of organically cultivated area in our country has reached the 110000 ha. The utilisation of organic seed is regulated by national law (82/2002. (IX. 4.) FVM-KvVM Regulation) According to this regulation the use of organic seed is obligatory in organic plant production from 01.01.2004. This regulation makes it important to find alternative methods for seed treatment instead of chemical treatment which is not allowed in organic farming. The situation is difficult because of the low amount of non-chemical seed treatment methods and additionally most of them are not effective enough. The goal of our research is to find methods, which keep good quality of organic seeds , assure healthy seed and safety field emergence. Experiment has two parts. In the first part the effect of the chosen materials were measured on the germination ability of the seeds. In the second part the effect of the chosen materials were observed on the seed-borne fungi of two vegetable plants. The two plants are green pea (Pisum sativum L.) and maize (Zea mays L.) The chosen materials are warm water, Biokál (permitted in ecological farming as plant conditioning substantial), Alginit and Vetozán (permitted in ecological farming as fertiliser) and oil of thyme. Ascochyta sp. on green pea and Fusarium moniliforme var. subglutinans on maize was observed as seed- borne fungi. Effects on germination ability were observed in the laboratory and in the field as well. Phytopathological tests were made in vitro after international rules (International Rules for Seed Testing 2003). According to our present results Biokál enhanced germination and thymol gave the best pathological effect. Experiment will be continued, data analysis is on. Technology will be available at the end of the experiment. Experiment is supported by OTKA T43072.

48 Budapest, Hungary, May 17th – 19th, 2004

41 - ALTERNATIVE CROSSES AND CRIS CROSS FOR MAIZE HYBRIDS AND SEED PRODUCTION IN NORMAL AND QUALITY PROTEIN MAIZE (QPM) ESPINOSA-CALDERÓN1, A., TADEO-ROBLEDO 2, M., SIERRA M3., M., SANDOVAL3, A., GÓMEZ M.3, N., BETANZOS M.3, E., COUTIÑO E3., B., CABALLERO H3., F., LÓPEZ-PEREIRA 4, M. & PIÑA D.V.5, A.

1Seed production and 3Maize Program, National Researches Institute of Forestry, Agriculture and Livestock (INIFAP, México), [email protected] 2, National University Autonomous of México (UNAM, México), [email protected] 4 Mexico Economy Program, CIMMYT until 1993. 5 Plant Breeder of CERES Seed Enterprises.

In the Plant breeding process the plant breeders define the conformation of the hybrids. However in some cases it is necessary to change the sequence of the conformation. The factors for interchange the lines order are good performance like female and male: 1) seed productivity, 2) seed quality, 3) flowering synchrony from female to male, 4) seed disponibility. In the single crosses the changes are not complicated because there are only two progenitors, but in Mexico this kind of hybrid is not usual. The three way hybrids now are the more usual kind of hybrid in Mexico for the National Researches Institute of Forestry, Agriculture and Livestock (INIFAP). The double crosses were very popular for three decades from 1950 until 1987. Many Quality Protein Maize hybrids, three way and single cross hybrids, were evaluated from 1997 until 2001. Some of the three way hybrids had highest yield that the tester but they have some problems to obtain certificated seed. We have evidences that will be better when the conformation of the progenitors is changed. In the case of the single crosses it is necessary to develop technology and seed production for increasing the yield of the progenitors, the less seed yield must be 2.5 tons per hectare. In this paper are showed some results with the criss cross method in normal and Quality Protein Maize (QPM) hybrids. We confirm that some hybrids need to change after to be released the progenitors order for improved the seed production process.

42 - LMPROVEMENT OF SUNNHEMP (CROTALARIA JUNCEA L.) SEED BY GRADING GHOSH, N., DAS, N.R. & GHOSH, M.

RC Agricultural University, B-Q/110, Kalyani Nadia – 741235, West Bengal. India

Sunnhemp (Crotalaria juncea L) is a leguminous bast fibre crop mainly cultivated in lndia, Bangladesh and Brazil. It is an ideal crop for uncultivated fallow or freshly reclaimed areas since it builds the soil fertility by fixing atmospheric nitrogen by rhizobium activity in root nodules. It is a short day plant, long day favour vegetative growth and reduce seed set. Seeds of 'Bidhan Shan’ (a self compatible variety of sunnhemp developed by the authors) along with parental line (self incompatible) were graded as large and small from ungraded seed lot. They were treated with Artificial Ageing (AA) by placing at 100% Relative humidity at about 28°C (Room temperature) Rate of decrease of root length was slow in large seeds after AA Moreover, large seeds had better storability and could be stored better with seed moisture content about 11% The vigour of large seeds with higher initial capital was expressed not only in vegetative and root growth but also in the reproductive parts of number of pods and seed yield. Bidhan Shan performed better than the parental line. Thus increase in the seed size at planting was beneficial in terms of plant growth and components of seed yield and seed quality including storability.

49 27th ISTA Congress – Seed Symposium

43 - SOWING AND HARVESTING TIME ON SEED QUALITY AND LEAF YIELD OF KALMEGH (ANDROGRAPHIS PANICULATA NEES.) GHOSH, N. & GHOSH, M.

B.C. Agricultural University, B-9/110, Kalyani, Nadia - 741235, West Bengal, India.

Kalmegh (Andrographis paniculata Nees.) is a medicinal herb. It is thermo and photo sensitive for flowering. The flowering starts when day length comes down below 11.5 hours and maximum temperature drops down to about 30° C. Pods have shattering habit and seeds have some dormancy. A full grown plant yields about 240 capsules, each containing about 10 seeds with 85% germination. Seeds are small, light in weight about 1.6 gm for 1000 seeds and yellowish in colour. Leaves have maximum medicinal value, but bitter in taste due to andrographolide. Plants are harvested 110-120 Days After Sowing. Seeds attained Physiological maturity at 28 Days After Anthesis (DAA); however, maximum vigour was with 30 DAA. Seed dormancy could be removed by treating with 0.5% potassium nitrate. Four times of leaf pluckings could be made if sown early in April, after 60 DAS at one month's interval. Leaf harvesting in younger stages of plant would promote more leaves to be developed. Leaf and seed both could be harvested from the same plant, without hampering seed quality even after four times defoliation of leaves.

44 - COLORING ALTERATION IN WHEAT FLOUR ASSOCIATED WITH TRICHOTHECIUM ROSEUM FUNGI. JACCOUD-FILHO, DAVID S., FERRARI, ROSELI A., CARVALHO, G. & ZAMPIER, IGOR

Universidade Estadual de Ponta Grossa, PR, Brazil, [email protected]

One of the main quality criterias for wheat flours used in the production of alimentary pastery is its coloring, which influences directly the coloring of the end products, such as bread, biscuits etc. During the processing of a sample of wheat flour in the Campos Gerais region, in the state of Paraná, it was observed (via Peckar’s test) that the sample’s coloring presented a dark appearance, resulting in its commercial depretiation and limited usage for the industry. Aditional tests carried out with a Minolta Spectrophotometer, which expresses the flour’s color, also indicated an alteration in color when compared to a flour considered normal. The wheat grains which originated the darkened flour sample and those which originated the flour sample with normal color were analysed after an incubation period of seven days via the Blotter Test Method (12h light / 12h dark, 22ºC). In the grains which originated the darkened flour sample, the presence of the Trichothecium roseum fungi was detected at the incidence of 37.5%, and in the grains which originated the flour sample with normal color this fungi was not detected.

45 - DRYING MAIZE SEED USING AMBIENT TEMPERATURE AT LOW RELATIVE HUMIDITY. KRZYZANOWSKI1, F.C., WEST2, S.H. & FRANÇA-NETO1, J.B.

1Embrapa - National Soybean Research Center, Caixa Postal 231, 96001-970, Londrina, PR., Brazil. ([email protected]. 2University of Florida - Agronomy Seed Laboratory, P.O. Box 110770, 32611, Gainesville, FL, USA. [email protected]

Drying is a prerequisite to maintaining maize (Zea mays L.) seed quality. New technologies are available and a dryer prototype has been developed that has the unique feature of taking the moisture out of the air stream before it is heated and passed through the seed. The heat that is generated when the water is removed is reintroduced into and raises the temperature of the air stream. In contrast to the 45°C temperature typical of conventional dryers, the 50 Budapest, Hungary, May 17th – 19th, 2004 temperature of the air stream in the new method does not exceed 35°C. No detrimental effects were observed on various parameters of mayze seed physiological quality due to the drying conditions provided by the heat pipe technology. The time required for drying was reduced significantly compared to the traditional system of heated ambient air. The heat pipe system is a promising technology to be used for drying shelled seed.

46 - DRYING PEANUT SEED USING AMBIENT TEMPERATURE AT LOW RELATIVE HUMIDITY. KRZYZANOWSKI1, F.C., WEST2, S.H. & FRANÇA-NETO1, J.B.

1Embrapa - National Soybean Research Center, Caixa Postal 231, 96001-970, Londrina, PR., Brazil. [email protected]. 2University of Florida - Agronomy Seed Laboratory, P.O. Box 110770, 32611, Gainesville, FL, USA. [email protected]

The moisture content of peanut kernel (Arachis hypogea L.) at digging ranges from 30 to 50% on a wet basis (w.b.). The seed moisture content must be reduced to 10.5% or below before they can be graded and marketed. After digging, peanuts are cured in the windrow for two to five days then mechanically separated from the vine. Heated air is used to further dry the peanuts from approximately 18 to 10% moisture content w.b. Drying is required to maintaining peanut seed and grain quality. Traditional dryers pass a high temperature and high humidity air stream through the seed mass. The drying time is long because the system is inefficient and the high temperature increases the risk of thermal damage to the kernels. New technology identified as, heat pipe technology (HPT) is available and has the unique feature of removing the moisture from the air stream before it is heated and passed through the seed. A study was conducted to evaluate the performance of the HPT system in drying peanut seed. The seeds inside the shells were dried from 17.4% to 7.3% in 14 hours and 11 minutes, with a rate of moisture removal of 0.71% mc per hour. This drying process caused no reduction in seed quality as measured by the standard germination, accelerated ageing and field emergence tests. It was concluded that the HPT system is a promising technology for drying peanut seed when efficiency and maintenance of physiological quality are desired.

47 - DRYING SOYBEAN SEED USING AMBIENT TEMPERATURE AT LOW RELATIVE HUMIDITY. KRZYZANOWSKI1, F.C., WEST2, S.H. & FRANÇA-NETO1, J.B.

1Embrapa - National Soybean Research Center, Caixa Postal 231, 96001-970, Londrina, PR., Brazil. ([email protected]. 2University of Florida - Agronomy Seed Laboratory, P.O. Box 110770, 32611, Gainesville, FL, USA. [email protected]

Under subtropical and tropical environments soybean seed (Glycine max (L.) Merrill) are harvested early to avoid deterioration from weathering. Careful after-harvest drying is required and is an important step in maintaining the physiological quality of the seed. Soybean seed should be harvested when their moisture content is in a range of 16-20%. Traditional drying utilizes a high temperature and high humidity air stream passed through the seed mass. The drying time is long because the system is inefficient and the high temperature increases the risk of thermal damage to the seed. New technology identified as heat pipe technology (HPT) is available and has the unique feature of removing the moisture out of the air stream before it is passed through the seed mass at the same environmental temperature. Two studies were conducted to evaluate the performance of HPT to dry soybean seed. In the first trial the seeds were dried from 17.5% to 11.1% in 2 hours and 29 minutes and in the second trial the seeds were dried from 22.56% to 11.88% in 16 hours and 32 minutes. This drying process caused no reduction in seed quality as measured by the standard germination, tetrazolium-viability, accelerated aging and seedling vigour classification tests. The only parameter that indicated a slight seed quality reduction was tetrazolium vigour in the second trial. It was concluded that the HPT system is a promising

51 27th ISTA Congress – Seed Symposium technology for drying soybean seed when efficiency and maintenance of physiological quality is desired.

48 - STUDIES ON SEASONAL EFFECT ON FORAGING BEHAVIOR OF HONEYBEE ON HYBRID SEED PRODUCTION IN BOTTLEGOURD PUSA HYBRID-3 KUMAR, SATISH, SINHA, S.N. & TOMAR, B.S.

Division of Seed Science Technology, Indian Agricultural Research Institute New Delhi-110 012, INDIA.

The hybrid seed production of bottlegourd (Lagenaria Siceraria M.)variety Pusa Hybrid-3 was taken up during Kharif and spring season of 2001 and 2002 at seed production unit farm, Indian Agricultural Research Institute, New Delhi. The planting of male and female rows were made in ratio of 1 : 4. During the Kharif season, honey bee begun their flight activity around 8 A.M. with highest flight activity at 12 noon and thereafter decline trends was observed in their flight activity (i.e at 2 pm, 4 pm and 6 pm). Among the honeybee species, Apis Millifera was observed to be dominated pollinating agent, where as Apis cerana was least observed as pollinating agent. During Kharif season A. Millifera spent more time (42.33 sec), followed by A. dorsata (28 sec), A. florea (12.33 sec). A. cerana (11.67 sec) and other pollinating insect (10.67 sec) at 12.00 noon. Whereas in summer season also the honeybee start their flight activity around 8 AM, attained peak at 12.00 noon and subsequently declined at 2 pm, but further gain their flight activity around 4 pm. The declined in flight activity at around 2 pm due to high temperature and hot dry winds. The time spent by the various pollinating agent viz., A. millifera (27.00 sec), A. dorsata (21.67 sec), A. florea (13.33 sec) and A. cerana (12.00 sec) other pollinating insect (10.67 sec.) at 12.00 noon was recorded in summer season. In Kharif season at 2 pm A. millifera spent (40.67 sec), A. dorsata (26.33), A. florea (13.67), A. cerana (15.67) other pollinating insect (7.67 sec) which was significantly higher than that of time spent by A. millifera (12 sec), A. dorsata (8.33 sec), A. florea (6.33 sec), A. cerana (8.6 sec) and other pollinating insect (8.33 sec) time at 2 pm in summer season. In summer season A. millifera spent (20.67 sec), A. dorsata (16 sec), A. florea (6.7 sec), A. cerana (10 sec) and other pollinating insect (8 sec) at 4 pm where as during Kharif season A. millifera spent (28.67 sec), A. dorsata (17.00), A. florea (11 sec). A. cerana (8.33 sec.), other pollinating insect (6.33 sec) time at 4 pm. It was revealed from this investigation that pollinating insect spent more time at 12.00 noon and 2.00 pm. in kharif season which synchronize with maximum stigme receptivity and pollen production , where as in summer season pollinating insect spent more time at 12.00 noon and at 4.00 pm. During Kharif season fruit yield per plant was 15.57 kg with no. of fruit, per plant, 7.61 which was higher than that of the fruit yield per plant 7.50 kg with no. of fruits per plant 5.33 in summer season. The seed yield per fruit 68.60 gm and 105.43 gm in summer and kharif season was recorded respectively.

49 - THE PERFORMANCE OF GLYPHOSATE AFFECTED BARLEY SEED MCLAREN, GILLIAN & RONALD DON

Official Seed Testing Station for Scotland, Scottish Agricultural Science Agency, East Craigs, Craigs Road, Edinburgh, EH12 8NJ, United Kingdom, [email protected]

The laboratory germination of barley seed from crops sprayed pre-harvest with glyphosate improved when tested in compost rather than paper towelling. However, field emergence and establishment were more closely related to the results of paper towelling germination tests than those from germination tests in compost. The effects of glyphosate are limited to the mother crop and once this is established they not transmitted to the following generation.

52 Budapest, Hungary, May 17th – 19th, 2004

50 - EFFECT OF FOLIAR FUNGICIDES APPLICATION ON SOYBEAN SEED QUALITY MOLINA, J. 1Y2 ; CARMONA, M.2.; BABBITT, S. 1Y2; GALLY, M.2, SUGIA, V.2 & GRIJALBA, P.2

1.Dirección de Calidad, ex INASE. Secretaría de Agricultura, Ganadería, Pesca y Alimentación. [email protected] 2.Facultad de Agronomía, Universidad de Buenos Aires.

Foliar fungicide application to control late season diseases of soybean (Glycine max) has been recently adopted in Argentina. However, the information about seed quality harvested from treated fields is poor. The aim of this study was to evaluate the effect of foliar fungicide application in reproductive stages of soybean, on seed weight and quality. Several diseases affect soybean crop in Pampeana Region and had been causing high damages lately, specially late season diseases that reduce yields and seed quality. Six trials were conducted in 2001-2002 growing season in south Santa Fe province in no-tilled monoculture commercial soybean fields. Treatments were arranged in a complete random block design, and consisted of one application of triazoles, benzimidazoles or strobirulins in reproductive soybean stages R3 or R5. The following variables were measured from harvested seeds: 1000 grains weight and germination in sand, by means of evaluation of normal seedlings, following the International rules for seed testing currently used (ISTA). Results indicated that 1000 grains weight was significantly higher in treated plots in all trials, with no differences between fungicides or time of application. Percentage of germination was related to seed infection level (Phomopsis spp, Cercospora kikuchii and Fusarium spp), and was significantly higher in seeds harvested from plots treated in R5 only in two trials

51 - PERFORMANCE OF DURA FAMILIES IN RELATION TO HYBRID SEED PRODUCTION IN OIL PALM P.MURUGESAN, P. & RETHINAM*, P.

National Research Centre for Oilpalm, Pedavegi-534450, Andhra Pradesh, India, [email protected]

Four dura families viz., 271D, 65 D, 120 D and 139 D were evaluated based on weight of fresh fruit bunch (FFB) and number of bunches. Effect of seasonal variation on these traits and height increment also were combined for evaluation. Seed garden raised during 1990 consisting selfed families (271D, 65D, 120D and 139D) were subjected for evaluation based on fresh fruit bunch yield, number of bunches, their seasonal variation in production and height increment. Based on initial five years yield, fourty four palms out of a total of four hundred palms crossed the standard of 150 kg of FFB /year/palm. The progeny of 65D and 120D had very less height increment, which registered an average height increment of 57.6 cm/year followed by 271D with 60.8 cm. They also had more number of bunches and yield. A peak and a lean period of bunch production were observed during the period May-October and November-April, respectively.

52 - EVALUATION OF SEED MATURITY OF CUCURBITS BY TETRAZOLIUM TEST NASCIMENTO1; W.W., PEREIRA1; R.S., COSTA1; C.J. & DIAS2, D.C.F.S.

1Embrapa Hortaliças, C.P> 218, 70359-970, Brasília, DF, Brazil; 2Department of Fitotecnia, Universidade Federal de Viçosa, 36571-000, Viçosa, MG, Brazil. [email protected]

The physical and physiological changes during cucumber and hybrid squash seed development under tropical conditions were investigated. This study was carried out in Brasilia, DF. Brazil, during the years of 2001 and 2002, using a cucumber cv. Eldorado 300 and a F1-squash hybride (Cucurbita maxima x C. Moschata) cv. Jabras. Fruits were harvested at 30, 40 and 50 days (cucumber) and at 20, 30, 40, 50 and 60 days (squash) after anthesis. Seeds were extracted from fruits after 0 and 10 days (cucumber) and after 0, 53 27th ISTA Congress – Seed Symposium

15 and 30 days (squash) of storage under packinghouse conditions. Dry seed weight, standard germination, first count germination, field emergence, and tetrazolium test (vigor and viability) were evaluated. For tetrazolium test, the seeds were imbibed in water at 40oC for 30 minutes for tegument removal, followed by 30 minutes for the internal membrane removal. Afterwards, they were exposed to a 0.075% tetrazolium solution for 60 minutes at 40oC. Five classes of viability and vigor were established, and each seed evaluated was allotted to one class. In cucumber, physiological maturity of seeds probably occurred at 50 days after anthesis; in general, the effect of fruit storage on physiological seed quality occurred in fruits harvested at 30 days after anthesis. For squash, seed physiological maturity occurred in fruits harvested between 40 and 50 days after anthesis and stored for 30 and 15 days, respectively. Seeds from these fruits had high vigor, germination and dry weight. Tetrazolium test was efficient to monitor seed quality changes of cucurbits during maturation. The major physical and physiological changes occurring during seed development and the importance of fruit storage post-harvesting are discussed.

53 - EFFECT OF IRRIGATION ON YIELD AND QUALITY OF SUGARBEET (BETA VULGARIS L.) SEED NOLI E., MONTANARI, M., ROSSI PISA, P., URSO, G., GRASSI, E. & LOVATO, A.

Department of Agroenvironmental Science and Technology, ‘Alma Mater Studiorum’ Università di Bologna,Viale G. Fanin 40, I-40127 Bologna, Italia

Sugar-beet seed production is one of the most remunerative seed production activities in northern Italy. Currently most Italian seed companies recommend irrigation in the period from flowering to seed filling, although published research on this topic is scarce. The objective of this work was to evaluate the effect of the scheduling of irrigations on sugar- beet seed yield and quality. Field trials were carried out in 1996, 1997 and 1998; transplants were performed at the end of February. Seven sprinkle irrigation treatments were tested, namely a at onset, b at mid, c at the end of flowering and their combinations a+b, a+c, b+c e a+b+c, and compared to a non-irrigated control. Each application depth ranged from 25 to 45 mm. Soil water content for each treatment was determined on samples taken from the first 40 cm of the soil profile before and after each irrigation. Moreover, in 1997 and 1998 samples were taken weekly, from the first irrigation until the harvest, at various depths. Irrigations affected water content in soil mainly in the first 20 cm which are those mostly explored by the root system and exposed to evapotraspiration. All irrigation treatments, carried at mid and at the end of flowering, either individually or in combinations, significantly increased seed yield in each year as compared to the non- irrigated control, whereas 1,000-kernel weight and germinability were unaffected. The percentage of seedballs of size greater than 4 mm responded to irrigation scheduling. Simple computations indicate that the extra costs for irrigation could be largely compensated by the higher yield obtained with respect to the non-irrigated crop.

54 - FIXING SEEDING INTERVAL IN RICE HYBRIDS PATIL, VEERANAGOUDA, BIRADARPATIL, N.K., SHEKHARGOUDA, M. & BIRADAR, D.P.

University of Agricultural Sciences, Dharwad -580 005, India

Synchronization in flowering between the parents in hybrid seed production and CMS multiplication is necessary to enhance the out crossing rate. The sowing dates of the parents should be adjusted to get simultaneous flowering. The growth duration of the parental lines varies with locations and seasons. The leaf number and the effective accumulated temperature requirements have to be worked out for each parental line. Hence, the study was taken up at two locations in Northern Karnataka region to find out the seeding interval for hybrid seed production of KRH-1, KRH-2 and DRRH-1 hybrids and CMS multiplication of IR 58025A and IR62829A. The growth duration and effective accumulated temperature requirement of the parental lines varied with location, where as the number of leaves produced by the parental lines was 54 Budapest, Hungary, May 17th – 19th, 2004 relatively stable. The number of leaves produced up to 50 per cent flowering ranged from 9.9 (IR9761-19-01R) to 15.7 (KMR-3). For taking up of seed production of KRH-2 and DRRH-1, the male sterile line (IR58025A) of these hybrids should be sown after their respective restorer lines (KMR-3 and IR40750R) attains the leaf number of 1.6 and 1.1, respectively. Where as, for KRH-1 seed production, the restorer line (IR9761-19-01R) should be planted after the male sterile line (IR58025A) attains the leaf number of 4.2. The parental lines, KMR-3 and IR9761-19-01R took higher and lower EAT of 1445.3oC, respectively for attaining 50 per cent flowering compared to other parental lines. The restorer lines of KRH-2 and DRRH-1 required higher EAT compared to their respective male sterile lines, where as the restorer line of KRH-1 required lesser EAT than the CMS line. The maintainer lines recorded lesser EAT compared to the male sterile lines

55 - EFFECT OF DESICCATION ON SOME QUALITY CHARACTERISTICS OF SUGARBEET SEED RAJIC, MILORAD, MARINKOVIĆ, BRANKO, MIKLIČ, VLADA & PANKOVIĆ, LAZAR

Institute of Field and Vegetable Crops, Novi Sad, Serbia and Montenegro, Faculty of Agriculture, Novi Sad, Serbia and Montenegro

A three-year field trial has been established in a system of random blocks in order to study the effect of desiccation on the 1000-seed mass and germinability of sugarbeet seed under the local climatic conditions. Desiccations were performed at the beginning and end of wax maturity. As the timing of cutting and threshing operations is important in the harvest of seed sugarbeet, desiccation simplifies the overall harvest. The application of desiccation under the local agroecological conditions has caused significant changes in the studied quality characteristics. The 1000-seed mass was significantly larger in the control than in the desiccation variants. The later application resulted in higher seed mass compared with the earlier application. Significant differences were also registered in seed germinability. It was therefore clear that the interruption of growing season negatively affected the major quality characteristics of sugarbeet seed. In view of the significant differences among the study years, the obtained results showed that seed sugarbeet responds uniformly to different climatic conditions.

56 - SEED QUALITY VARIATION WITHIN CEREAL SEED LOTS AND ITS DETERMINANTS SCHMOHL, SVEN & KRUSE, MICHAEL

Universität Hohenheim (350D), Institute of Plant Breeding, Seed Science and Population Genetics, Fruwirthstr. 21, 70593 Stuttgart, DE-Germany

Heterogeneity of seed lots is permanently under discussion. The present ISTA maximum seed lot sizes as well as the sampling schemes are directly linked to this basic quality trait of seed lots. But experimental data about variation in seed lots are quite rare. Computer simulations were made for identifying optimum sampling schemes but are based on assumptions about the variation within seed lots without being directly confirmed by practical data. The aim of the present project is to quantify in detail the variation of the important seed quality traits purity, other seed count, germination, thousand seed mass and seed size within commercial seed lots of wheat, barley, rye and oats. Between 50 and 60 primary samples were taken from the raw material by an automatic seed sampler at the delivery point of a seed plant and also after each step during seed processing up to the bagging device. In addition, primary samples were taken with a cargo sampler from 37 positions in the seed lots during each intermediate storage in a silo. All primary samples were analysed separately. The presentation of the results will focus on the absolute level of variation as well as on the changes of the level and type of variation during seed processing. It will highlight the importance of seed cleaning for the decrease of variation and the separation effects during filling and emptying a silo.

55 27th ISTA Congress – Seed Symposium

These detailed results shall be used to define an efficient sampling protocol for estimating the variation in seed lots in other seed plants and in other species to establish a representative data base on the variation within seed lots. This data base may then be used for re-calculating the sampling schemes and to re-assess the maximum lot size as a measure for avoiding excessive heterogeneity.

57 - IDENTIFICATION OF DIAGNOSTIC CHARACTERISTICS IN SOYBEAN VARIETIES SHAILENDRA SINGH, M.P.

State Seed Certification Agency, Office Complex, Gautam Nagar, BHOPAL PIN- 462023, MADHYA PRADESH ( INDIA )

Studies were carried out in Soybean with varieties (JS-81-335, JS-90-41, PK-1024 and JS- 93-05) to verify Identification characters at seed and plant levels, in different locations in Madhya Pradesh (India). The Varieties were verified based on distinguishing morphological characters at seed level; the varietal characters observed at seed level such as seed coat colour, seed shape, hilum shape and seed lusture were most stable and uniform. These identified seeds were planted in the field and were compared with the authentic varietal characters. It was observed that the varieties identified at seed level were true to the type in the field also. At field level; the varietal characters like foliage colour, flower colour, growth habit, pigmentation and hairiness were observed the most stable, uniform distinguishing characters for verification of soybean varieties. Although some other characters viz; days to flowering, days to maturity and plant height were the uniform characters but has poor stability due to interaction with genotype with environment for expression. The off type plants were identified on the basis of these characters. Some other important points which also need to be taken care of are mentioned below: Off type and other uprooted plants should be properly removed. Seed Crop should be inspected timely and critically.

58 - STORAGE OF THE COFFEE SEEDS (Coffea arabica L.): DRYING METHODS AND ENVIRONMENTS VIEIRA1; A. R. GUIMARÃES2; R.M. OLIVEIRA2; J.A. PEREIRA2; C.E. & CARVALHO2. F.E.

1EPAMIG, CP 176, CEP 37200-000, Lavras-MG, [email protected]; 2UFLA, CP 37, CEP 37200-000, Lavras-MG, [email protected]

The loss of seed viability is one of the greatest problems faced by coffee seed producers during seed storage. Thus, the performance of coffee seeds (Coffea arabica L. cv. Rubi) during storage harvested in 2001/2002, at Federal University of Lavras-Brazil (UFLA), was investigated. After cleaning seeds were kept at 47,5% of water content (WC), fast (forced air chamber at 35oC) and slow (open environment with 20oC and 60% of relative humidity (RH)) dried until reaching 12% WC. The seeds were packed in hermetic bags and stored for 9 months at two environments: regular, with monitored temperature and RH, and in dry cold chamber at 10oC and 50% of RH. Every three months seed quality was evaluated by the germination test, seedling emergence, emergence speed index, tetrazolium test, phenols determination and biochemical analyses by using electrophoresis of the enzymes alcohol dehydrogenase, catalase, endo-β-mannanase, malate dehydrogenase, peroxidase, poliphenoloxidase and superoxide desmutase. Fast drying decreases the vigor of seeds stored at dry and cold chamber environment, but viability is only affected after six months of storage. In a regular warehouse environment the vigor is affected but viability starts decreasing after the third month of storage. Such results are confirmed by the changes detected at the level of phenols and enzymes profiles studied.

56 Budapest, Hungary, May 17th – 19th, 2004

59 - A NEW OLD SPECIES: TRITICUM DICOCCUM ZECCHINELLI, RITA, FERRARI, FABIO, BARBIERI, GIOVANNA, FUSARI, M. LAURA, MALLOZZA, ELISABETTA, MAZZOLA, PAOLA & SPELTA, ROSSELLA

ENSE – Laboratorio Analisi Sementi, Via Emilia km 307, 26838 Tavazzano LO, Italy TEL +39 0371 761919 FAX +39 0371 760812, [email protected]

Triticum dicoccum Schrank ex Schubl. is a traditional crop of some Italian regions and in particular of Garfagnana, in Tuscany. In the last years, this old wheat has been rediscovered, in the framework of the development of organic farming and the “fashion” of “natural food”. Consequently, seed samples of this new old species – not included in the current ISTA Rules – reach today the Italian Seed Testing Stations, asking investigations about the laboratory methods to verify the quality of the seed lots to be marketed. Triticum dicoccum seems very similar to another species of the same genus, Triticum spelta and both have hulled seeds, while important differences are to be considered and first of all the number of chromosomes (T. dicoccum is tetraploid, T. spelta is hexaploid). The morphological similarity of these two species is a difficulty to be faced by purity analysts, but the attention at some particular characteristics of the caryopsides helps their identification. Germination tests can be carried out using the methods prescribed for the other Triticum spp, but – as in T.spelta – the analysts have to take into account the multiple units that frequently occur. The poster with pictures and drawings wants to present Triticum dicoccum, its seeds and seedlings and to compare it with Triticum spelta in the aim to introduce this species to the colleagues, as the introduction in the ISTA Rules of Triticum dicoccum is in progress. The proposed testing methods are also presented.

57 27th ISTA Congress – Seed Symposium

3. Viability and Vigour: Evaluation and Impact Topics: Methods of evaluation; influence of seed quality on seed performance and/or emergence, storage potential, responses to stress

60 - GENETIC VARIATION FOR CARDINAL TEMPERATURES OF GERMINATION IN WHEAT FROM THE NORTH OF IRAN AKRAM GHADERI*., FARSHID, SOLTANI , AFSHIN, SADATI, JAFAR & ZEINALI, ABRAHIM

Department of Agronomy, Gorgan University of Agricultural Sciences, Gorgan, Iran *[email protected]

This study was conducted to determine genetic variation, broad-sense heritability and expected genetic advance for cardinal temperatures of germination and tolerance ranges to temperature in genotypes of wheat from the north of Iran. Base, optimum and maximum temperatures for germination, responsibility to temperatures lower than optimum and responsibility to temperature higher than optimum and tolerance range in temperature lower than optimum, higher than optimum and total tolerance range in 12 genotypes of wheat from the north of Iran were analyzed. The genotypes did not exhibit significant differences for all the traits, except for responsibility to temperatures lower than optimum. Broad- sense heritability ranged from 81 for responsibility to temperatures lower than optimum to 11 % for tolerance range in temperatures lower than optimum. Also, expected genetic advance ranged between 7.7 for responsibility to temperatures higher than optimum to 0.01% for maximum temperature. With respect to low estimates of genetic variation and expected genetic advance for traits evaluated (expect for responsibility to temperatures lower than optimum), for breeding wheat cultivars for the traits it may be necessary to introduce genotypes from other locations or cross programs may be used.

61 - EVALUATION OF PERCENTAGE OF GERMINATION, TOTAL SPEED OF GERMINATION AND VIGOUR INDEX OF 17 MEDICINAL PLANTS SPECIES TO AGEING TEST ALIZADEH, MOHAMMAD ALI

Scientific Board member of Research Institute of Forest and Rangeland, P.O. Box 13185-116, Tehran, Iran, [email protected]

The objective of this research experiment is to determination and evaluation of percent of germination, total speed of germination and vigor index of the seed of 17 species of medicinal plants. (According to the International Seed Test Association, 1985) different tests including (Standard germination test) with normal condition and accelerated ageing test condition were used for determination of percent and total speed of germination of 17 species of medicinal plants. The vigor index was calculated by following equation: percent of germination * the total mean of length of root and shoot and its result was followed with the results of percent of germination and total speed of germination. Among those species, some species were resist to the accelerated ageing test and some species were not, nevertheless the percent and speed of germination, of those seed samples which is resist to accelerated ageing, are meaningful, comparing with non-aged seeds.

58 Budapest, Hungary, May 17th – 19th, 2004

62 - PHYSIOLOGICAL QUALITY OF SCHINOPSIS BALANSAE ENGL. (QUEBRACHO COLORADO) SEEDS ALZUGARAY, C.1, CARNEVALE N. J.2, SALINAS, A. R. 2 & PIOLI, R. 1

1Facultad de Ciencias Agrarias. Universidad Nacional de Rosario (UNR). Campo Experimental Villarino. C.C.14. S2125ZAA. Zavalla. Santa Fe Argentina. 2 Consejo de Investigaciones de la UNR (CIUNR). Facultad de Ciencias Agrarias. Universidad Nacional de Rosario (UNR). Campo Experimental Villarino. C.C.14. S2125ZAA. Zavalla. Santa Fe Argentina. [email protected] and [email protected]

S. balansae constitutes the dominant species in the forests that occupy the North of Santa Fe Province, Argentina. The goal of this work was to evaluate the physiological quality of S. balansae seeds. Trials were done with seeds harvested in 2000 and 2001. The seeds were evaluated at the 3rd, 12th, and 18th months of the harvest of the seeds. It was evaluated the viability by the Tetrazolium test and the Standard Germination test; and the vigour by the germination velocity index (GVI). It was evaluated the incidence of pathogenous in the samaras and in the seeds. At the 3 months of harvesting, the seeds showed 77% of germination in 2000, that diminished to 29% at the 18 months of harvest. Seeds harvested in 2001, with 28% of initial germination, practically lost their viability at the 18 months. The GVI showed the greatest vigour of the seeds on the 3rd month of the harvest in 2000. Pathogenous associated to the lowering of the germination were found. The viability and the vigour tests were good indicators of the quality of the seeds of S. balansae. The low viability of the seeds in 2001 seemed to be caused by adverse environmental conditions.

63 - ASSESSMENT OF GENETIC PURITY, IDENTITY AND SEED VIGOUR IN F1 HYBRIDS AND F2 POPULATION IN COTTON ANKAIAH, R., RAMANADHAM, R., REDDY, N.M., KESHAVULU, K. & REDDY, B.M.

National Seed Project, ANG Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, India, [email protected]

Experiments were conducted to characterize cotton genotypes and to correlate electrophoretic banding pattern of total soluble seed protein with genetic purity by field grow-out test (GOT). Key morphological characters, which were least influenced by growing conditions were identified for hybrids and their parents and these characters were found distinct, uniform and stable (DUS characters) throughout the growing season. Further, key characters for recombinants in F2 segregating population were identified and these recombinant plants were considered for identification of F2 seed lots. Based on morphological characters, flow chart was developed which is useful for field functionaries involved in seed production/certification for easy identification of off types. Grouping of cotton genotypes could be possible based on differential growth response of seedlings to added chemicals. The electrophoretic banding patterns of total soluble seed proteins and globulins, each genotype was unique and distinct between hybrids and their parents. These differences were useful in identification of individual cotton genotypes. The genetic purity results obtained by field plot technique (GOT) and electrophoretic technique were not correlated. Hence, field GOT is found to be foolproof method for genetic purity testing. The study on yield and yield attributes revealed significant reduction of yield in F2s over F1s due to reduction of boll number, boll weight and number of seeds per boll and 100 seed weight. Use of F1 seed is always beneficial to farmers to reap 15 to 20 per cent higher yield over parents and 35.8 to 38.5 per cent over F2 seed.

59 27th ISTA Congress – Seed Symposium

64 - EFFECT OF GENETOYPES AND CONTAINERS ON THE STORABILITY OF SOYBEAN SEED ANKAIAH, R., MANOHAR REDDY, N. & MURALIMOHAN REDDY, B.

National Seed Project, ANG Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, India, [email protected]

Soybean seed cv. MSCS 58, MSCS 13, HARDEE, KBSH 2 having an initial germination of 80% and 7.5% moisture were packed in polythene bag, poly-coated bag and cloth bags and stored under ambient conditions of Rajendranagar, Hyderabad, India. Observations were taken on germination and vigour index at bi-monthly intervals. The germination percentage and seedling vigour index were high in seed stored in polythene bag (700 guage) and poly- coated bags in all genotypes. The storability of soybean seed could be extended by 6 months in cloth bags and 14 months in polythene bag provided the initial moisture content at the time of storage is less than 8%.

65 - EFFECT OF FUMIGANT ON RICE SEED VIABILITY, VIGOUR AND NATURAL INSECT INFESTATION ANKAIAH, R., MANOHAR REDDY, N. & MURALIMOHAN REDDY, B.

National Seed Project, ANG Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, India, [email protected]

Rice seed of Samba Mahsuri var. BPT 5204 having 92% germination, 9.4% moisture and 1.5% natural insect infestation initially was stored in four metal bins (24 guage) separately which was subjected to fumigation with celphos (aluminium phosphide) once, twice and thrice and kept for storability under ambient condition of Rajendranagar. Storability was recorded in months as the period up to which seeds had shown above 80% germination (certification standard). The natural infestation of insects and total fungal colonies were less in fumigated seed as compared to un-fumigated seed (control). Control seed had 14 months storability only. Fumigated seed for once, twice and thrice had storability with more than 80% germination up to 12, 16, 14 and 14 months respectively. Fumigated seed twice and thrice recorded lesser seedling vigour index during storage period. Thus fumigation of rice seed with less than 10% seed moisture once and stored in metal bins of 24 guage was found effective for controlling insect damage and improving shelf life of seed up to 16 months.

66 - EFFECT OF VAPOUR PROOF CONTAINERS ON SEED STORABILITY IN ONION ANKAIAH, R., MANOHAR REDDY, N. & MURALIMOHAN REDDY, B.

National Seed Project, ANG Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, India, [email protected]

Onion seed of N 53 cv. having 7.2% moisture and 80% initial germination was dried to 4.98% seed moisture in silica gel desiccator and stored in cloth bag, paper bag (moisture pervious containers), polythene bag (700 guage), aluminium foil pouch, cikatainer and 150 gauge poly pouch (vapour proof containers) under ambient conditions of Rajendranagar, Hyderabad, India. Observations on germination, seedling vigour and moisture content were made at bi-monthly intervals. The germination percentage and seedling vigour index were high in seed stored in aluminium foil pouch and polythene bag throughout the storage period when compared with other containers. Seed stored in cloth bag, paper bag and poly pouch had only 4 months storability with more than 70% germination. Seed stored in polythene bag and aluminium foil pouch exhibited 75% germination up to 18 and 10 months respectively. Among the vapour proof containers, aluminium foil pouch was found very effective for extending the shelf life of seed from 4 to 18 months.

60 Budapest, Hungary, May 17th – 19th, 2004

67 - A METHODOLOGY TO DEVELOP SEED VIGOUR TESTS ARTOLA¹, A. & CARRILLO-CASTAÑEDA², G.

¹INIA. Programa de Semillas. La Estanzuela. Colonia. Uruguay. C.C. 39173. [email protected]).²Colegio de Postgraduados. IREGEP-Genética. Montecillo. Estado de México. México. CP 56180. [email protected]

High quality seed lots produce a rapid and uniform establishment of plant stand that maximizes yield potential under a wide range of soil conditions. Seed vigour testing is now a common practice for quality control involved in seed production, conditioning, storage and marketing processes. A methodology was developed with the purpose to develop seed vigour tests, which comprises three steps. Calibration: Two seed lots from a cultivar with similar germination percentage but with different seed vigour are used. A stressing condition is selected and the precise experimental condition under which the two lots express the greatest difference in seed germination is determined. Preliminary evaluation: Several seed lots, characterized to have sufficient variability in physiological quality, are subjected to the test and the laboratory data are compared with data of seedling emergence in soil. Validation: It is according to ISTA method validation programme for seed testing. Two seed vigour tests were developed in Lotus corniculatus L. by performing seed germination under low temperature (5ºC) and vacuum stress conditions. In the calibration, the biggest discriminatory power to differentiate seed lots for its vigour condition was achieved at 7 days in the vigour test at 5ºC; and at 72 h and 8.7 psi of pressure in the vacuum test. The preliminary evaluation, sampling 10 seed lots allowed verifying their discriminatory capacities to classify seed lots according to its potential planting value because both seed vigour tests showed a high correlation (r = 0.95 - 0.96**) with seedling emergence in soil. This potential methodology deserves further investigation.

68 - STUDIES ON INFLUENCE OF SEED VIGOUR ON INITIAL SEED QUALITY AND YIELD OF OKRA (ABELMOSCHUS ESCULENTUS) ARUN KUMAR, M, KESHAVULU, KUNUSOTH, MANOHAR REDDY, NALLA, ANKAIAH, RAGULAKOLLU & MURALIMOHAN REDDY, BHOOMIREDDY

Dept. of Seed Science & Technology, National Seed Project, Acharya N.G. Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, A.P., India, [email protected]

The studies on influence of seed vigour on initial seed quality and yield of okra (Abelmoschus esculentus L.) was carried out during post rainy (Rabi) season of 2002-03. Freshly harvested seeds of okra hybrids (Sun 008 and Uphar) were subjected to accelerated ageing at 40 ± 1°C and 85 ± 5% relative humidity for 4, 8, 12 and 16 days for inducing different seed vigour levels measured in terms of seed germinability. The resultant seed lots had 84, 76, 65 and 55 per cent germination in the hybrids Sun 008 and 82, 75, 66 and 57 per cent germination in the hybrid Uphar. The two hybrids with four vigour levels constituted the treatment aspect of this investigation. The resultant effect of lower vigour levels on initial seed quality decreased seed germination and seedling length (22.57 – 14.25 cm and 23.12 – 14.00 cm in Sun 008 and Uphar respectively), dry weight of the seedling (2.14 – 1.40g and 2.20 –1.44 g in Sun 008 and Uphar respectively), seedling vigour index (2138.7 – 770.2 units and 2098.5 – 794.52 units in Sun 008 and Uphar respectively) and exhaustive vigour tests (43.75 – 26.25 units and 39.00 – 22.25 units in Sun 008 and Uphar respectively). Plants established from low vigour lots exhibited lesser field emergence index (80.49 – 44.02 units and 79.36 – 47.45 units in Sun 008 and Uphar respectively), shorter plant height (85.29 – 53.27 cm and 87.42 – 56.83 cm in Sun 008 and Uphar respectively), low dry matter production per plant (50.41 – 30.85 gm and 50.55 – 31.42 g in Sun 008 and Uphar respectively), smaller leaf area per plant (346.52 – 241.99 cm2 and 353.22 – 245.24 cm2 in Sun 008 and Uphar respectively), decreased number of green fruits (31.00 – 20.00 and 33.00 – 22.00 in Sun 008 and Uphar respectively) and seeds per fruit (46 – 33 and 47 – 34 in Sun 008 and Uphar respectively) and seed yield (33.66 – 15.70 g and 33.92 – 13.94 g in Sun 008 and Uphar respectively). Growth parameters, yield components and seed yield had positive significant correlation with vigour levels. The regression analysis indicated that for every one per cent decrease in germination percentage, there was a reduction of 267 g and 281 g of green fruit yield per plot (20 sq.m) in Sun 008 and Uphar respectively. 61 27th ISTA Congress – Seed Symposium

69 - EFFECTS OF COTTON SEED CHARACTERS ON GERMINATION AND EMERGENCIES IN LABORATORY AND FIELD CONDITION AZAD DISFANI, F. & ZANGI, M.R.

Board of scientific, Cotton Research Institute of IRAN, [email protected]

One of the primary objectives of cottonseed production programs is to provide farmers with seeds having a high potential for germination and establishment of a good, uniform stand of seedlings on field condition. 8 cotton genotypes were evaluated. Genotypes were 43259, Siokra 324, Sahel , Sindose, Varamin, Backtegan, Gockrova and 818-312.Different seed characters were evaluated. Characters were seed length, seed width, seed weight, seed volume, cotyledon weight, germination rate, and normal seedling number and emergences rate. Results showed that relationship between germination rate and all of traits (excepted seed volume) was non significant. Correlation between seed volume and germination was significant(r=0.39, p>0.05). Relationships between The Different seed characters and emergences rate was non significant.

70 - EVALUATION OF SEEDLING GROWTH, GERMINATION AND VIGOR TRAITS IN COTTON BARZALI1, MOHAMMAD, TAHMASEBI1, Z., HAJIREZAEI2, M.R. & BOERNER2, A.

1Tarbiat modarres university, POX 14155-7863, Tehran, I. R. Iran. 2Institute for Plant Genetics and Crop Plant Research (IPK), Gatersleben. Germany, [email protected]

In order to evaluate early seed vigour and germination traits in cotton, 26 various varieties were tested in the field for different parameters. Four varieties (Delta pine 50, Red leaf okra, Shirpan 603 and Oltan) with the most seedling dry matter were selected for a detailed analysis under in-vitro conditions. Seed reserve utilization efficiency, germination capacity, germination velocity, germination rate index, bulk density, coleorrhiza length, coleoptile length, seedling length, total dry matter, coleoptile dry matter and colerrhiza dry matter were used for analysis after 7 and 12 days in germinator and incubator, respectively. The results showed that there were significant differences among seed reserve utilization efficiency, germination capacity, germination rate index, coleorrhiza length, coleoptile length, seedling length, total fresh weight, coleorrhiza dry weight and coleoptile dry matter (P<%5). In comparison to other three varieties, red leaf okra produced highest seedling dry matter at twelve days after cultivation in an incubator and contained the most seed reserve utilization efficiency, germination capacity and germination velocity (P<%5). Furthermore, coleoptile length was reduced in red leaf okra compared to Delta pine 50. There was a significant correlation between dry matter production and coleoptile’s dry matter as well as total fresh weight. Possible explanations for differences in seed vigour and germination traits are presented and discussed.

71 - SEED QUALITY ANALYSIS IN HYBRID RICE DURING STORAGE AND FIELD EMERGENCE BASNAL, GOPAL SINGH

Indo-American Hybrid seeds (I) Pvt. Ltd. Bangalore, India

Seeds of two rice hybrids IR-58025AX Pusa-150-9-3-3R and IR-58025AX Suweon 318R and their parents: cytoplasmic male sterile (CMS) line IR-58025A, its maintainer IR-58025B and restorer lines Pusa-150-9-3-3R and Suweon-318R with checks, Pant Dhan-4 and Pant Dhan- 10 were stored in cloth bags under ambient conditions of Pantnagar, India for 8 months. Germination, seedling length and seedling vigour index increased during early part of the storage. Later on, a gradual decrease in these characters was observed. The maximum

62 Budapest, Hungary, May 17th – 19th, 2004 decrease was observed in hybrids followed by CMS line, its maintainer and restorer lines and lowest in check varieties. The maximum decrease in protein content was observed in maintainer line (0.6%) and lowest in restorer Suweon–318R (0.1%). The highest decrease in starch content was observed in CMS line (1.8%) and the lowest in restorer lines (0.6%). On the other hand the highest increase in electrical conductivity of seed leachate and leaching of water-soluble sugars was found in CMS line followed by hybrids, maintainer, restorer lines and check varieties. The highest decline in α-amylase activity was observed in CMS line and the lowest in restorer lines. It was positively correlated with percent field emergence (r =0.929), field emergence index (r =0.928) and seedling fresh weight (r =0.778). Field emergence and its emergence index were higher in check varieties and restorer lines than in hybrids and other parents. However, seedling height, seedling fresh and dry weight were higher in hybrids and followed by checks and restorer lines than CMS line and its maintainer.

72 - STUDIES ON SEED SIZE IN INFLUENCING GERMINATION, YIELD AND QUALITY OF SOYBEAN, WHEAT AND MUNGBEAN BASNAL, GOPAL SINGH

Indo-American Hybrid seeds (I) Pvt. Ltd. Bangalore, India

Seeds of all the three crops divided into four group according to the size and sown in the field. Field emergence was highest with medium sized seed in soybean however, in case of wheat and mungbean field emergence increased with an increasing seed size. Lowest emergence was observed with small size seed in all the crops tested. Whereas seed yield per plant is positively correlated with seed size in all the crops tested. In case of soybean highest yield per hectare was observed with medium sized seed and lowest with smallest size seeds. In case of wheat and mungbean highest seed yield observed with boldest sized seed. and lowest with smallest size seed. Same trend was observed for processed seed yield also. Test weight of final processed seed increased with increasing seed size in all the test crops. It means sowing of boldest seed produced heaviest seed. Germination test was also done in next generation, the germination % was observed with medium sized seed in soybean but highest with boldest size seed in mungbean and wheat. Whereas all other quality parameters such as germination index, shoot length, root length, seedling dry weight, seedling vigour index, were higher in boldest size seed and lowest in smallest sized seed. Thus it can be summarized that Medium sized seed (5.0-5.5mm dia.) in soybean and bold sized seed (>3.0mm dia.) in Wheat and (>3.35mm dia.) in Mungbean performed best for germination and plant stand in the field, which ultimately resulted in highest seed yield of good quality. Vigour parameters such as germination index, shoot length, root length and seedling dry weight were found best with boldest size seed in all the test crops. It is suggested that medium size seed or comparatively smaller sized seed in soybean and boldest size seed in wheat and Mungbean may be basic in achieving higher yield levels of quality seed.

73 - CORRELATIONS BETWEEN THE SIZE AND SHAPE OF HYBRID MAIZE (ZEA MAYS L.) SEEDS AND YIELD COMPONENTS BERZY1, TAMAS, HEGYI1, ZSUZSANNA, ZABORSZKY2, SANDOR & PINTER1, JANOS

1Agricultural Research Institute of the Hungarian Academy of Sciences, Martonvásár, [email protected] 2 Georgikon Faculty of Agriculture, Veszprém University, Keszthely

Introduction Much research has been published on the importance of the genetic hereditary material, in the present case hybrid maize seed. The production of hybrid maize seed plays an especially

63 27th ISTA Congress – Seed Symposium important role in agriculture in Hungary, quite apart from the breeding institutes and the hybrids they develop. In addition to satisfactory cultivation techniques, plant nutrition and pest control, the most important component in achieving high yields is the use of seed with excellent biological value. The biological value of the seed can be characterised by adaptation to unfavourable early environmental conditions, early vigour, and the development of a satisfactory plant stand under field conditions. It has long been known that there are differences in quality between seed fractions of various shapes and sizes (Barla-Szabó 1984, Berzy 1982, Eisele 1981, Fiala 1977, Germ 1966, Kietreiber 1972, Thielebein 1958), in favour of the flat fractions. Earlier research in Hungary (Pásztor 1962, Heiczmann 1964), however, found no difference in the biological value of the seed from the point of view of yield and recommended reducing the number of fractions. Seed production may be endangered by a number of stress factors. Under such conditions different responses may be given by seed of various shapes and sizes (Krishnaveni and Vanangamdi 1989, Scotti and Godoy 1978, Kushibiki and Kummahata 1980, Martinelli and Carvalho 1998), which can be attributed to differences in the biological value of the seed. Mention should also be made of the pests and pathogens which threaten the initial development of the seedlings (Roundhawa et al. 1990). The present studies were carried out on state registered maize hybrids, the seed of which was divided into size fractions in order to examine the biological value of the seed under laboratory and field conditions, based primarily on the yield components.

74 - GERMINATION, VIGOUR AND RESPONSE TO SIMULATED WATER DEFICIT AT GERMINATION OF HULLED AND HULLESS SPRING BARLEY. BOROS, LECH & KOLASIŃSKA, K.

Department of Seed Science and Technology, Plant Breeding and Acclimatisation Institute at Radzików, 05-870 Blonie, Poland, [email protected]

Plant emergence and establishment is a crucial issue in farming system. Poor establishment can be a problem in covered barley but is a frequent and more serious problem in hulless barley. Conditions during seed development and maturation, mechanical damage during threshing and subsequent handling (aggressive seed coat removal, embryo bruising and cracking and even embryo removal from the seed) of the harvested seed are considered to be a very important cause of poor seed and seedling vigour. The objective of this study was to recognise the factors causing lower seed quality of hulless in comparison to hulled barley after harvest and processing as well as to compare the response of contrastive barley genotypes to simulated water deficit at germination phase. The basic seed of Rataj (hulled) and Rastik (hulless) cultivars from 2002 harvest were used in these studies. Seed quality assessment was done after pre-processing and in spring after processing. Seeds of both cultivars harvested with plot combine and adjusted to moisture content of 10 and 15% were also tested for germination at reduced water potential using PEG 6000 solution. Seed germination of hulled cultivar was 88% with 9% of abnormal seedlings, whereas hulless 85% with 7% abnormal seedlings. The accelerated aging (AA) test proved comparable high seed vigour of both hulless and covered barley. The processing and storage conditions caused the significant decrease of sowing value of hulless seeds. Averaged across initial seed moisture content and water stress, the hulless cultivar Rastik had greater germination energy (GE), percentage of abnormal seedlings (PAS), shorter mean germination time (MGT) and higher germination index (GI) although germination percentage (GP) of controls of the two cultivars was similar. Initial seeds moisture significantly affected energy of germination and percentage of germination. All osmotic potential treatments markedly reduced EG and GP as compared with control. Mean germination time increased along with reduced water potential, while the opposite relationship was noticed for germination index. The hulless Rastik produced seedlings with longer plumule and shorter roots comparing with covered one. Shoots growth was more sensitive to reduced water potential than roots independently of initial seed moisture content. Increasing water deficit progressively decreased plumule and root length with higher reduction observed for hulless barley. The results have confirmed that hulless barley is more sensitive to mechanical damage during threshing and handling is more sensitive to storage conditions and to water

64 Budapest, Hungary, May 17th – 19th, 2004 deficit during germination. There are other agronomic factors like fungicide treatment, deep seeding that may further aggravate germination problem.

75 - THOUSAND SEED WEIGHT OF BARLEY SEED AND ITS RELATIONSHIP TO GERMINATION CADGER, C. & HALL, J.

Scottish Agricultural Science Agency, East Craigs, Edinburgh, EH12 8NJ, United Kingdom, [email protected]

Thousand seed weight of barley is a varietal characteristic but it can vary from year to year depending on climatic conditions. In practice in any given year there are significant differences between varieties and within varieties, the thousand seed weights of individual seed lots can vary by more than 100%. There is no relationship between thousand seed weight and germination, and growers should use germination and thousand seed weight information in determining the weight of seed required to achieve their target seedling population.

76 - A SEED VIGOR TEST FOR Lotus corniculatus L. BASED ON VACUUM STRESS CARRILLO-CASTAÑEDA¹, G., ARTOLA², A. & DE LOS SANTOS¹, G. GARCÍA

¹Colegio de Postgraduados. IREGEP. Montecillo, Estado de México. México. CP 56230. [email protected] ²INIA. Programa de Semillas. La Estanzuela. Colonia. Uruguay, C.C. 39173. [email protected]

Lotus corniculatus L. displays deficient crop establishment due to its low seed vigour; therefore, it is recommended to carry out seeding with optimum vigour seed. The development of a practical method, with the discriminating capacity to rank seed lots in terms of seed vigour is presented. Two seed lots of birdsfoot trefoil cultivar San Gabriel with similar germination percentages, one harvested in 1999 (SG-A) and the other in 1993 (SG- B), were exposed to vacuum conditions during germination process. The greatest difference (P ≤ 0.05) in germination between the two seed lots was observed when the seed was exposed for 72 h to 8.7 psi negative pressure (vacuum) at 20 C. The germination of seed, exposed to these conditions, was highly significantly correlated (P ≤ 0.01) with seedling emergence in soil (r=0.92-0.99**). The discriminatory capacity of the vacuum method was subsequently demonstrated using 10 seed lots. The vacuum test differentiated between seed lots according to their vigour condition, with germination ranking from 67 to 7%, which reflected their potential planting value. This test requires only 3 days for seed vigor evaluation, and the method is more precise than the standard germination test for determining physiological quality among seed lots. It also complies with the basic requirements for its standardization. The method described here has been named the vacuum test (VT).

77 - THE GENETIC DIVERSITY IN WILD POPULATIONS OF PROSOPIS LAEVIGATA IN THE HIDALGO STATE, MÉXICO. PRESERVATION OF THIS NATURAL RESOURCE CARRILLO-CASTAÑEDA1, GUILLERMO & JUÁREZ MUÑOZ2, JUANA

1Colegio de Postgraduados. IREGEP. Montecillo, Méx. 56230. México, [email protected] 2Universidad Autónoma del Estado de Hidalgo. Centro de Investigaciones Forestales. Tulancingo, Hgo. 43600. México.

Nearly 70% of the Mexican territory, classified as arid, is known to support nine species of mesquite (Prosopis spp.) which are an important natural resource in this country. P. laevigata is used for building materials, fuel, fodder and honey. Taxonomic classification of 65 27th ISTA Congress – Seed Symposium

Prosopis species has been hindered by morphological similarities. In an attempt to address this problem, we used random amplified polymorphic DNA (RAPD) markers to compare the genetic diversity of natural populations of P. laevigata from the Mexican State of Hidalgo. Two groups of phenotypes related by similarity coefficients of 0.92 were identified. Four and five subgroups per group were identified and a total of 9 different phenotypes identified. These results give evidence for genetic diversity within and across the two populations studied. A selection of seed from the 9 different phenotypes identified are been studied from the point of view of seed germination and by their potential planting value. This study is required to generate in the glasshouse the plants of the selected phenotypes in order to forest or re-forest wide arid areas. In simple terms, it is possible to affirm that seed vigour is the process opposite to deterioration. The greatest seed vigour is obtained at physiological maturity and from this moment seed deterioration starts. One of the manifestations of seed deterioration is the loss of cell membrane integrity, which reduces the ability of the seed to retain solutes. We have developed important techniques to improve the physiological quality of seed and to differentiate seed by their potential planting value across a wide range of soil types and environmental conditions. A priming approach is a strategy to restore the germination capacity of seeds. This treatment significantly reduced the time to 50% of seed germination. Similarly, the time to 30% emergence is reduced and in general, the uniformity of both germination and seedling emergence are also considerably improve and seed vigor is increased. The vacuum test differentiated seeds according to their vigour condition, with different germination ranking, which reflected their potential planting value. This test requires only 3 days for seed vigor evaluation, and the method is more precise than the standard germination test for determining physiological quality in seed lots. It also complies with the basic requirements for its standardization. Results of the performance of P. laevigata seeds from the 9 different phenotypes identified shall be presented.

78 - EVOLUTION OF SEED QUALITY DURING THE FRUIT DEVELOPMENT ON SUGAR BEET MOTHER PLANT CASALS1, MARIE-LAURE, LADONNE1, FABIENNE & NARDI2, LOUIS

1FNAMS, Impasse Le verger - 49800 Brain/Authion – France 2FNAMS, La haille, Route de Nérac – 32100 Condom - France

To get homogeneous field emergence and growth plant, sugar beet producers have very strict requirements regarding to seed germination. A number of physical factors have been found to influence the germination and establishment of sugar beet seed for example low temperatures, restriction of oxygen uptake, moisture of substrate. Nevertheless, low germination may also be related to the maturity of fruit. Over two years, the French Seed Grower Union (FNAMS) has conducted field trials to examine sugar beet fruit and seed development during maturation and their relationships germination ability. The plants used, in the experiments, consisted of a monogerm pollinators and a monogerm cytoplasmic malesteriles. 60 plants were tagged, at one day intervals, on the same branches (using twists of coloured plastic). All these stem sections presented flowers with the same age (flowers open out the same day). At six dates after flowering (from 14 to 60 days after anthesis), stem sections were harvested, dried in controlled conditions and stored at low temperature until the germination test. The results show that germination occurred only on seeds harvested between 15 – 20 days after anthesis (DAA) and reached the maximum on seeds harvested after more than 40 DAA. Seed and fruit weight increased until about 40 DAA. Different hypothesis to explain the evolution of the seed germination are proposed.

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79 - STUDY OF RELATIONSHIP BETWEEN LABORATORY SEED VIGOUR TEST AND FIELD PERFORMANCE IN DIFFERENT GENOTYPES OF GREEN GRAM (VIGNA RADIATA L. WILEZEK) CHAKRABARTI, S.K.

Seed Science and Technology Department, Faculty of Agriculture, Bidhan Chandra Krishi Viswavidyalaya (Mohanpur), Pin-741252 (W.B.), INDIA

Eight genotypes of green gram (Vigna radiata L. Wilczek) were studied during the year 2002 and 2003 to find out relationship of laboratory vigour test and field performance parameters and to specify one or two laboratory vigour test to predict field performance. The field experiment was conducted at the Instructional Farm during pre-kharlf season (February to May) in randomized block design. The experimental data indicated particularly two laboratory vigour test viz. electrical conductivity and accelerated aging tests are dependable for the above purpose. The experimental results showed a negative and significant relationship between electrical conductivity and field stand at maturity. Average yield per plot was negatively and significantly associated with electrical conductivity. Accelerated aging bad positive and significant association with field stand at maturity and plot yield. The correlation study indicated that by going through the results of electrical conductivity and accelerated aging, it is possible to predict field stand at maturity and also the seeds which are stored can be evaluated for quick selling or further storage. This investigation result can help farmers as well as seed testing laboratories and seed marketing agencies for quality ranking of their stored seeds.

80 - EVALUATION OF GERMINATION CAPACITY IN DIANTHUS GLACIALIS, AN ENDANGERED ALPINE SPECIES COLOMBO1, A., CASTIGLIONI1, A., TOSCA1, A. & BONOMI.2, C

1Fondazione Minoprio - Centro Regionale per la Tutela della Flora Autoctona, Viale Raimondi, 54 - 22070 Vertemate con Minoprio - Como – Italy, [email protected] 2Museo Tridentino di Scienze Naturali, Via Calepina, 14 - CP393 - 38100 Trento – Italy

Experiments described are part of the research project “Ex situ conservation: Seed Banking Trentino Endangered Species”. To maintain a genetic diversity representative of the population sampled, a method to achieve complete germination of all viable seeds is needed. In the case of rare plant from which only small seed samples could be collected, this goal is even more necessary. Therefore seed viability and germinability have been tested on wild seed lots of Dianthus glacialis. Tetrazolium analysis has been set up. Germination data were recorded every second day till the 54th day after the test begun. Percentage of seed germination, germination index and mean time to germination have been calculated for priming treatments with gibberellic acid A3 at different concentrations from 0 to 0.72 mM. Seeds primed for 48 h with Gibberellic acid at 0.58 mM reached 31% of germination as the non treated test. Water treated test and lower concentrations of gibberellic acid gave poorer results. Germination index was higher for non treated test and the concentration of 0.72 mM of gibberellic acid. Mean times to germination were from about 6 to 9 day depending upon the treatment, but no statistical difference were assessed for this parameter. It is supposed the Dianthus glacialis does not tolerate priming for 48 h and it could benefit of shorter time of priming treatment.

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81 - EVALUATION OF THE PHYSIOLOGICAL QUALITY IN COFFEE SEEDS (COFFEA ARABICA L.), CULTIVARS ACAIÁ, BY ELECTRICAL CONDUCTIVITY TEST. COSTA, P.S.C. & CARVALHO, M.L.M.

Universidade Federal de Lavras, DAG, CP 37 - CEP 37200-000 Lavras/ MG – Brasil, [email protected]

The conductivity test is used to measure the leaches from the seeds, which indicate the membrane system integrity. The test is rapid, inexpensive and it is of simple operation. However, the methodology needs to be tested for each individual species in order to reach a better precision and accuracy. This work was conducted to verify the physiological quality of the coffee seeds, with and without parchment, through the electrical conductivity test using the individual and the bulk methods. The second goal of this work was to determine the imbibition time and the partition point in that test. Ten seed lots of C. arabica, cultivars Acaiá were used in this study. The imbibition curve for each lot was determined and the seeds were submitted to the germination and tetrazolium tests. For the evaluation of the partition point seeds were imbibed for 96 hours and submitted to the germination test correlating the conductivity values observed with the physiological quality. The individual and bulk methods proved to be efficient in distinguishing coffee seed lots from different levels of physiological quality. The imbibition’s period of 96h for individual method and seeds without parchment were the most appropriate conditions for the development of the conductivity test. The partition point is 120,5mS/cm, considering 70% of germination.

82 - EVALUATION OF THE PHYSIOLOGICAL POTENCIAL OF BEET SEEDS DA SILVA, JOSUÉ BISPO & VIEIRA, ROBERVAL DAITON

UNESP, Jaboticabal, Depto. De Produção Vegetal, UNESP/FCAV, 14884-900 JABOTICABAL, SP, Brazil, [email protected], [email protected]

The use of an adequate vigour test is a very important tool to evaluate the physiological potential of produced and commercialised beet seeds. This research was carried out in order to study the procedures of the accelerated ageing and controlled deterioration tests as vigour test for beet seeds as well as to relate its results to seedling field emergence. Seven beet seed lots of “Top Tall Early Wonder” cultivar were submitted to standard germination, accelerated ageing (42oC per 24, 48 and 72 hours) and controlled deterioration (41 and 45oC; 22, 24 and 26% of seed moisture content; 12, 24 and 36 hours of ageing period). Based on the results it can be concluded that the accelerated ageing test might be carried out at 42oC per 72 hours while the combination of 45oC/24%/24h might be used as a procedure for controlled deterioration test. Then, the vigour of beet seeds can be evaluated using the accelerated ageing and controlled deterioration tests, with the mentioned combinations.

83 - REFINING THE TETRAZOLIUM TEST AND ITS ANALYSIS FOR SEED CONSERVATION COLLECTIONS OF GRASSES AND OTHER WILD PLANT SPECIES. DAVIES, HANNAH, E., ALI, NATASHA, S., HOYLE, GEMMA & PROBERT, ROBIN J.

Seed Conservation Department, Royal Botanic Gardens, Kew, Wakehurst Place, Ardingly, West Sussex, RH17 6TN, UK, [email protected]

Tetrazolium tests (TZ) on 108 seed collections representing 80 species across 12 tribes of wild grasses held at the Millennium Seed Bank were compared with the best estimate of viability from germination tests. A simplified approach to the analysis of TZ was employed. Seeds were allocated to one of three possible staining categories: 1) embryos uniformly well stained = viable. 2) embryos unstained = non viable. 3) embryos patchy or faintly stained =

68 Budapest, Hungary, May 17th – 19th, 2004 uncertain. Approximately 50% of the tests had seeds in the latter category. These were interpreted to be low vigour individuals with accumulated damage. In the absence of specific guidelines for the interpretation of staining patterns an arbitrary assumption was made that 50% of such individuals would be capable of germinating; accepting that this will be an underestimate in some cases and an overestimate in others. Using this approach, estimates of viability were not significantly different for 81 collections. In 26 collections TZ gave a significantly higher estimate of viability suggesting the presence of dormant individuals whereas in only 1 case did TZ under estimate viability. ISTA recommends that grass caryopses should be either dissected transversely below the embryo or dissected longitudinally through the embryo in preparation for TZ staining. The two methods were compared directly for 34 seed collections, representing 32 species. On average the transverse dissection gave a better match to the germination estimate of viability. Estimates were not significantly different for 24 collections when the transverse dissection was applied compared to only 15 for the longitudinal dissection. Advantages and disadvantages of amended TZ methods and analysis for a wider range of families are discussed.

84 - APPLICABILITY OF THE TOPOGRAPHICAL TETRAZOLIUM TEST FOR SEED VIABILITY TESTING OF WILD PLANT SPECIES DAWS, MATTHEW I., TERRY, JANET, MILES, STEPHANIE, DAVIES, HANNAH & WOOD, CHRISTOPHER B.

Seed Conservation Department, Royal Botanic Gardens, Kew, Wakehurst Place, Ardingly, West Sussex RH17 6TN, UK, [email protected]

The topographic tetrazolium test (TZ test) is widely used for rapidly assessing seed viability in a wide range of species, particularly those that have been recently harvested, are dormant or are slow to germinate. However, there are only International Guidelines for the use of TZ for 87 mainly agricultural, horticultural and forestry species. For these species, the test has been validated and embryo staining patterns interpreted. However, for other species there are concerns about the applicability of the test; results can be complex and subjective. TZ is widely used for seed viability testing with species for which there are no International guidelines on methodology and the interpretation of staining patterns. For example, the TZ is routinely used within the Millennium Seed Bank Project for viability testing of wild plant species. Here we examine the wider applicability of the test by presenting data comparing viability estimates (TZ test) with germination levels for >200 wild plant species. TZ provided an accurate assessment of viability for many species. In particular, it was useful for species with dormancy i.e. low recorded germination but high estimated viability. Thus for many species the TZ test may enable a differentiation between dormancy and seed in- viability. However, for some species, particularly in the Solanaceae and Aizoaceae, germination vales were consistently higher than viability estimates. Thus, for these species the test as currently implemented does not work and points to the need for further research to optimise either staining times, or embryo dissection prior to incubation in TZ.

85 - RECOMMEDATIONS FOR GERMINATION TEST PROCEDURES FOR NATIVE TREE SPECIES FROM THE BRAZILIAN ATLANTIC FOREST. DE S. MEDEIROS, A.C., DE A. ABREU, D.A., DOS SANTOS, A.F. & SOUSA, V.S.

Embrapa Florestas – National Forest Research Center of EMBRAPA, Cx. P. 319, 83411-000, Colombo, PR – BRAZIL, [email protected]

Embrapa Florestas set up a native tree seed bank - BASEMFLOR established in Colombo, State of Paraná, Brazil, in order to provide seeds for uses such as reforestation programs, reclamation of degraded ecosystems, establishment and restoration of riparian forests and forest covers on water catchment areas, botanical gardens, local and regional seed banks, and for research projects. One of this research projects involves development of seed technology including studies for breaking dormancy and germinating seeds. Seed morphology was studied, as well, when it was considered to help in devising appropriate treatments to promote germination. Recommendations for germination test for “vacum” 69 27th ISTA Congress – Seed Symposium

(Allophylus edulis – Sapindaceae), “cataia” (Drimys brasiliensis – Winterceae), “tamanqueiro” (Aegiphila sellowiana – Verbenaceae), ”angico-pururuca” (Anadenanthera falcata - Mimosaceae), “guaçatonga” (Caesaria sylvestris – Flacourtiaceae), “pixiricão” (Miconia cabucu – Melastomataceae), “angico-branco” (Anadenanthera colubrina – Mimosaceae), “canudo-de-pito” (Escallonia montevidensis - Saxifragaceae); “peroba-rosa” (Aspidosperma polyneuron - Apocynaceae), “vassoura-vermelha” (Dodonea Viscosa – Sapindaceae), “erva-mate” (Ilex paraguariensis – Aquifoliaceae); “bugreiro” (Litharaea brasiliensis – Anacadiaceae) and “baguaçu” (Talauna ovata – Magnoliaceae) were defined.

86 - FITTING TEMPERATURES PROFILES TO CALIBRATE THERMIC REGULATION PROBES FROM CLIMATIC ROOMS USED IN SEED TESTING DEMILLY, DIDIER, BILLY, BÉATRICE, HERBERT, OLIVIER, WAGNER, MARIE HÉLÈNE & LECHAPPE, JOËL

GEVES SNES Rue Georges Morel 49071 Beaucouzé Cedex, France, [email protected]

Several seed tests as germination or seed health must be performed in climate rooms or incubators. Temperature conditions and associated tolerances are described in the ISTA methods e.g. 20°C+/-2°C for the germination tests. To fulfil these requirements it is necessary to characterize thermostatic chambers (temperature profiles) and to calibrate the probes used to check the temperature. Laboratories which are applying for accreditation against the ISO standard 17025:1999 are frequently required by their national accreditation bodies to determine and use uncertainty of measurement for the probes and the temperature profiles. In our laboratory, germination rooms are constantly monitored with temperature probes in one point in the room. A method has been developed to ascertain the temperature profile of equipment, to calibrate simultaneously the probe according to the profile and to determine the uncertainty. This work was based on the national norm AFNOR XPX 15 140 may 1999 “Measurement of air moisture – Climatic and thermostatic chambers – Characterization and verification”. A calibrated temperature monitoring equipment was used with 9 probes. For each room temperature has been recorded in 9 points at least. Several parameters as mean, variability, uncertainty and conformity to the expected temperature have been calculated. They allow to check if the temperature regulation is well adapted, and to specify correction and acceptable limits to be applied to the probe used for routine check. Acceptable limits can be determined taking account of the whole uncertainty from both temperature profile and check probe.

87 - STORAGE POTENTIAL OF COTTON SEEDS PREDICTED BY VIGOUR TESTS AND BIOCHEMICAL ASSAYS. DIAS, D. C. F.S., FREITAS, R.A., DIAS, L.A.S. & OLIVEIRA, M.G.A.

Department of Fitotecnia / BIOAGRO, Universidade Federal de Viçosa - UFV, 36571-000, Viçosa, MG, Brazil, [email protected]

This study was done to determine the ability of biochemical assays and vigour tests to predict the storage potential of cotton seeds. Cotton seeds of IAC-20 RR and Fabrika cultivars were stored from May 2001, for 12 months under ambient environmental conditions and in a cool room (13°C) in Viçosa, MG, Brazil. Seed quality was assessed before storage by standard germination, accelerated aging, cool germination and electrical conductivity tests and biochemical assays (lipoxygenase and acid phosphatase activity, trypsin inhibitor, lipid and acid content). The generalized Mahalanobis distances procedure was used to assess the relationship between vigour tests and biochemical assays carried out before storage and the values obtained in the standard germination test performed after 2, 4, 6, 8, 10 and 12 months storage. There was the greatest similarity or smaller and no significative Mahalanobis distances among standard germination, accelerated aging and cool germination results. It was observed that accelerated aging and cool germination test performed before storage estimated seed viability after 8 and 10 months storage under environmental conditions, respectively. Electrical conductivity test and biochemical assays were not efficient to predict storability of cotton seeds. 70 Budapest, Hungary, May 17th – 19th, 2004

88 - INCIDENCE OF THE SUBSTRATE ON THE GERMINATION OF SUNFLOWER SEEDS DUCOURNAU1, S., WIESNER2, L., GREGOIRE3, S. & LECHAPPE1, J.

1GEVES – SNES, Rue Georges Morel, BP 24, 49071 Beaucouzé Cedex, France, [email protected] 2USDA-NCGRP, 1111 South Mason Street, Fort Collins, CO 80521-4500, USA 3GEVES, La Minière, 78285 Guyancourt Cedex, France

Use of compost is not allowed for germination test as primary substrate in ISTA and AOSA Rules. Several seed testing laboratories performed duplicate germination tests for Sunflower, including a test on compost, due to better germination of sunflower with this type of substrate. In order to decide if compost can be used as primary substrate for sunflower germination, a comparative test involving 15 ISTA and AOSA laboratories has been organised to compare germination of sunflower with compost, sand and paper. Germination tests have been performed on 5 samples of Sunflower, at the temperature of 20°C, with the 3 types of substrates. The ISTA method validation program has been used from the planning of the elaboration of the protocol. Statistical analysis shows that results of normal seedlings are higher with the use of compost for germination. This is due to a decrease of abnormal seedlings (principally) and of non germinated seeds (to a less extent). Repeatibility and reproducability of the results are also better with compost. Compost gives higher or equal results with the method used in routine in each laboratory, even if the laboratory is not experienced with compost. On the contrary, sand or paper can lead to lower results if the laboratory is not used to it. Results obtained in this experiment allow the recommendation of compost as primary substrate for Sunflower germination. A precise definition of compost will be proposed to be included in ISTA Rules.

89 - ACCELERATED AGEING AS A VIGOR TEST FOR SQUASH AND PUMPKIN SEEDS DUTRA, ALEK SANDRO & VIEIRA, ROBERVAL DAITON

UNESP, Jaboticabal, Depto. De Produção Vegetal, UNESP/FCAV, 14884-900 JABOTICABAL, SP, Brazil, [email protected], [email protected]

The accelerated ageing (AA) test is widely used as vigour test for many species among those are the vegetables crop ones. This research was conducted in order to study the adequate procedures to run the accelerated ageing as vigour test for squash and pumpkin seeds. Five seed lots of squash (“Menina Brasileira” cultivar) and five of pumpkin (Barbara Hybrid), with commercially accepted quality (germination) were used. To reach the objectives the standard germination, first counting and accelerated ageing ( traditional) and with salt concentrated solution) tests were run in laboratory. For AA test the combination of temperatures (38 and 41oC) and periods of ageing (48, 72 and 96 hours) were used. For each combination (temperature and ageing period) the seeds were place in a germination box, with 40 ml of deionized water, over a screen using a single layer of seeds. The salt concentrated solution was prepared using 40 g of NaCl and 100 ml of water. The results of AA test, both procedures, allowed identifying seed lots with high and low vigour. The ageing period of 41oC provided higher level of deterioration than 38oC, especially when was used 96 hours as ageing period. Based on the results it can initially be concluded that the combination of 41oC and 96 hours of ageing period should be used as a vigour test for squash and pumpkin seeds, however further studies have to be run in order to investigate others combinations and to confirm this results.

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90 - EVALUATION OF SEED VIGOR IN IRANIAN ELYMUS HIPIDUS ACCESSIONS ESMAILI, SHARIF MASOUD & KHAYAM, NEKOUEI MOJTABA

Isfahan Research Center of Agriculture and Natural Resources, Department of Seed Technology, Isfahan Iran, [email protected]

In order to improve and recover the pastures, growing of perennial grasses such as Elymus hispidus (opiz) Melderis is very important. However environmental stresses and lack of seedling vigor frequently prevent plants from emergence and growing in pasture. In this research, important components which effect on seedling emergence were investigated in laboratory, field and pasture. Fifteen accessions used in this study including, 10 accessions of E. hispidus seed from rangelands and five accessions during five stage from the farm were collected. Seed vigor tests were consisted of: Thousand Seed Weight (TSW), Electrical Conductivity (EC), Tetrazolium test (TZ), Standard Germination test (SG), Cold Soil test (CS), Cool Germination test (CG), Accelerated Aging test (AA), Seedling Growth Rate test (SGR) and Drought Stress test (DS). Seed accessions were planted in Fozveh Research Station (FRS) under irrigated condition and Chadegan Research Station (CRS) under dryland condition. The results indicated significant differences among seed accessions in terms of seedling emergence. Effect of planting date on seedling emergence percentage was significant. Data analysis showed all tests except EC and TZ detected vigor differences among seed accessions. Emergence percentage on CRS was significantly at 0.05 level with r- value of 0.73 and 0.71 correlated with germination percentage on 1st day from SG and CS tests respectively. Data analysis of TSW, SG, AA, CG, CS, SGR and DS tests were correlated with percent of seedling emergence on FRS. CS test had the highest correlation coefficient with seedling emergence under both conditions.

91 - GERMINATION OF ERVA-BALEEIRA SEEDS (CORDIA CURASSAVICA (JACQ.) ROEM. & SCHULT. - BORAGINACEAE). FANTINATTI, J.B., AGUIAR, R.H., CARNEIRO, L.M.T.A. & GROTH, D.

FEAGRI/Campinas State University (UNICAMP), Box 6011, Postal Code 13083-970, Campinas-SP, Brazil

Almost all the forest medicinal natives species of Brazil, are still found in wild state and growing naturally. It is necessary to know the physiological and morphologic aspects of seed growing in order to commercialy grown the plants. Cordia curassavica (Jacq.) Roem. & Schult. (Boraginaceae), known as erva-baleeira, it is a Brazilian native specie of commercial and medicinal importance. The main goal of this research was to carry germination tests of Cordia curassavica, that can be used in seed testing laboratory and reforestation programs. For the evaluation of seeds just-harvested and with 68,1% of moisture content and seeds dried at 40oC until 10,8% of moisture content, germination tests were conducted in towel paper and growed in constant temperature of 25°C and alternate of 20-30°C, with fotoperiod of 8 hours light and 16 darkness. It was evaluated the germination rate, index of germination speed (IVG) and the average time of germination (T50). The germination temperature and moisture contents, did not affected the percentage of germination, but it was shown a reduction in the index of germination speed and the average time of germination at 25oC.

92 - GERMINATION AND DORMANCY ELIMINATION IN SOME MEDICINAL PLANTS FARHOODI1, ROOZBEH, SHARIFZADEH, F., MAKIZADEH, M. & ESMAEILZADEH, H.

University of Tehran, [email protected]

According to increasing importance of medicinal plants cultivation, this research was conducted. These plants have some inhibiting factors of germination which don’t germinate easily infield and in order to crop those plants we need some mechanisms to stimulate them

72 Budapest, Hungary, May 17th – 19th, 2004 to germinate. The main objective of this study was to determine the best treatment for germination and dormancy elimination in laboratory on following plants: Calendula officinalis, Lavandula angustifolia, Echinacea purpurea, Chrysanthemum cinerrariae folium and Nepta caturia. This research was conducted base on CRD with 4 replications in seed laboratory using following treatments GA3, KNO3, stratification, physical and chemical scratch. The results showed that there are significant differences between treatments, some of those plants had good germination under GA3 treatments and some of them had good germination under KNO3 treatment and some of them had germination under stratification treatment and finally using suitable treatment has led dormancy elimination also promote germination and finally guarantee the growth of seedling in field.

93 - THE USE OF THE ACCELERATED AGING TEST FOR ESTIMATING THE POTENTIAL OF SEEDLING EMERGENCE IN THE FIELD OF SOYBEAN SEED LOTS FRANÇA-NETO, J.B.1, KRZYZANOWSKI, F.C1, PÁDUA, G.P.2, HENNING, A.A.1 & COSTA, N.P.1

1Embrapa Soybean, Caixa Postal 231, CEP 86001-970, Londrina, PR, Brazil; [email protected]; 2EMBRAPA/EPAMIG, Caixa Postal 351, CEP 38001-970 Uberaba, MG, Brazil

The accelerated aging test (AAT) is used for estimating a vigour index of seed lots and also for estimating their storage potential. For soybean seeds an exposure period of 72 h at a constant temperature of 41ºC and 100% RH is recommended by ISTA. However very few studies which relate the results of AAT with soybean seedling emergence in the field (SEF) are available in the literature. Preliminary results obtained by Embrapa Soybean indicated that the AAT could be used for estimating SEF, if the test is applied to soybean seeds just before planting time, and the stress period is reduced to 24 h. The objective of this study was to verify the possibility of using the data of vigour provided by the AAT (41ºC/100%RH/24h) for predicting the potential of soybean SEF. Regression analyses were obtained with the results of vigour provided by the AAT with the ones of SEF during the period of five years, from 1998 to 2002. A total of 879 seed samples of different cultivars were evaluated in the study. In each of the study year the AAT with an exposure period of 24 h was performed in October and in November (normal planting period for soybeans in Brazil) the evaluations o SEF were performed. All seed samples were treated with fungicide before planting in the field. The evaluations of SEF were done on the experimental field of Embrapa Soybean, in Londrina, Brazil. The regression analyses provided adjusted equations between the results of SEF and AAT. These analyses were individually evaluated for each study year and in conjunction for the five-year period. Comparing the individual equations obtained each year, it was observed that they were similar to each other. The conjunction analysis for the five-year period provided a regression equation between SEF and AAT (SEF = 0,6316AAT + 28,922; r2 = 0,83 ***), which was also similar to the individual equations obtained each year. This equation was applied to all AAT results of 100 seed samples evaluated in 2002. The values of SEF estimated by this equation were correlated with the actual recorded data of SEF, resulting in a coefficient of regression of 0.80, which was highly significant (P < 0.001). This equation was considered reliable for estimating SEF of soybean seed samples in soils of Londrina, with soil temperature and moisture levels near to ideal conditions.

94 - THE USE OF THE TETRAZOLIUM TEST FOR PREDICTING THE POTENTIAL OF SEEDLING EMERGENCE OF SOYBEAN SEED LOTS FRANÇA-NETO, J.B.1, KRZYZANOWSKI, F.C1, PÁDUA, G.P.2, HENNING, A.A.1 & COSTA, N.P.1

1Embrapa Soybean, Caixa Postal 231, CEP 86001-970, Londrina, PR, Brazil; [email protected]; 2EMBRAPA/EPAMIG, Caixa Postal 351, CEP 38001-970 Uberaba, MG, Brazil

The tetrazolium test (TTZ) provides a number of seed quality parameters, such as vigour (TZ-Vig), viability (TZ-Viab), and indexes of mechanical damage, field weathering, and stinkbug damage. The soybean seed-producing sector has demanded research studies that relate the results obtained by this test with the potential seedling emergence in the field (SEF). This type of information will greatly contribute for increasing the precision of soybean 73 27th ISTA Congress – Seed Symposium seed plantability. Additionally it will improve seed quality evaluation just before they are commercialised. The objective of this study was to verify the possibility of using the data of viability and vigour provided by the TTZ for predicting the potential of soybean SEF. Regression analyses were obtained with the results of vigour and viability provided by the TTZ with the ones of SEF during the period of six years, from 1997 to 2002. A total of 1,117 seed samples of different cultivars were evaluated in the study. In each of the study year the TTZ was performed in October and in November (normal planting period for soybeans in Brazil) the evaluations o SEF were performed. All seed samples were treated with fungicide before planting in the field. The evaluations of SEF were done on the experimental field of Embrapa Soybean, in Londrina, Brazil. The regression analyses provided adjusted equations between the results of SEF and TZ-Vig and TZ-Viab. The regression analyses were individually evaluated for each study year and in conjunction for the six-year period. Comparing the individual equations obtained each year, it was observed that the ones relating SEF and TZ-Vig were similar to each other. However this trend was not observed for the data relating SEF and TZ-Viab. This fact means that the TZ-Vig data is more reliable than those from TZ-Viab for estimating of SEF. The conjunction analysis for the six-year period provided regression equations between SEF and TZ-Vig and also TZ-Viab. Due to the fact that the individual equations for each year were more coherent and repeatable for TZ-Vig than for TZ-Viab, it was concluded that SEF is more precisely estimated using TZ-Vig information as reference. The obtained equation for the six-year period involving SEF and TZ-Vig (SEF = 0,6165TZ-Vig + 35,716; r2 = 0,73 ***) was applied to all TZ-Vig of 100 seed samples evaluated in 2002. The values of SEF estimated by this equation were correlated with the actual recorded data of SEF, resulting in a coefficient of regression of 0.79, which was highly significant (P < 0.001). This equation was considered reliable for estimating SEF of soybean seed samples in soils of Londrina, with soil temperature and moisture levels near to ideal conditions.

95 - EFFECTS OF HYDRATION AND DEHYDRATION PERIODS LENGTH ON THE COLEOPTILES AND ROOT GROWTH AND SEEDLING SURVIVAL OF 3 WHEAT CULTIVARS AFTER REHYDRATION GHORBANI, M.H., ZEINALI, E. & SOLTANI., A.

Dept. of Agron., Gorgan Univ. of Agric. Sci., Gorgan., Iran, Email: [email protected]

Wheat (Triticum aestivum L.) seedlings may meet dehydration between seed imbibition and emergence, particular in dryland farming. This laboratory experiment was conducted to study the effects of hydration and dehydration periods length on the coleoptile and root growth, seedling survival, and their ability to resume growth (after rehydration) of 3 wheat cultivars. Seeds of Golestan. Khazar-1 and Tajan cultivars were germinated for 4, 6 and 8 days in suitable moisture conditions. Then, seedlings were dehydrated for 2, 4 and 6 days. Number of normal seedling, lengths of coleoptiles and root, and dry weights of seed residues were measured after 7 days. Results showed that when seedlings were grown under suitable moisture conditions for 4 days, dehydration periods had no effect on the number of normal seedlings after rehydration, but an increase in the time of dehydration resulted to decrease the coleoptile and root length of 3 cultivars. Dehydration reduced normal seedlings and root lengths, significantly, when seedlings were placed under normal conditions before dehydration for 6 days or more. Reduction in coleoptile lengths as affected by dehydration was significant only for the treatment 8 days growth under suitable moisture conditions before dehydration. Dehydration period length had no effect on dry weights of seed residues in treatment 4 days growth under suitable moisture conditions before dehydration, while when seedlings were grown under suitable conditions for 6 days, increasing dehydration time followed by significant increase in dry weights of seed residues. In 8 days growth under suitable conditions before dehydration, increasing dehydration period length caused significant increase in seed residues only in Golestan cultivar. These results demonstrate that dehydration tolerance reduces as wheat seedlings develops. On the other hand, the length of dehydration can be effective on the ability of regrowth after rehydration. So this ability decreases as the length dehydration period increases

74 Budapest, Hungary, May 17th – 19th, 2004

96 - STORABILITY OF TOSSA JUTE (CORCLLORUS OLITORIOUS L.) SEEDS GHOSH, N., DAS, N.R. & GOSH, M.

Faculty of Agriculture, B.c.Agril. University, B-9/110, Kalyani, Nadia-741235, West Bengal, India.

Tossa jute (Corchorus olitorious L.) is a short day plant and commercially cultivated for bast fibre during long day. Seeds are small in size and orthodox in nature, without any dormancy. The experiment was conducted with five varieties of tossa jute using Artificial Aging test to determine inheritance pattern of storability. The result indicated that it was controlled mainly by Dominance, Additive and Epistatic components. Cytoplasmic genes also influenced the storability of tossa jute seeds, large seeds have better storability than small sized ones. Thus there is scope for improving storability of tossa jute seeds by selection. Storing in low temperature with low moisture contents of seeds at low Relative Humid Condition gave better result.

97 - EFFECT OF DIFFERENT GROWING CONDITIONS, PLANT DENSITIES AND YEARS ON THE QUALITY OF MAIZE (ZEA MAYS L.) SEEDS HEGYI, Z., KIZMUS, L. PINTÉR, J., BERZY, T. & MARTON, L.C.

Agricultural Research Institute of the Hungarian Academy of Sciences, Martonvásár

The aim of the experiments was to investigate the effect of different locations, stand densities, years and cytoplasm on the germination ability of maize seed. It is important for hybrid maize seed breeders to be able to estimate field emergence on the basis of laboratory germination tests (Gáspár 1968, Woodstock 1968). The germination percentage achieved in the laboratory should be seen as the theoretical maximum value of field emergence. The relatively few papers that have been published on the effect of the growing area on seed germination ability (Russell and Teich 1967, Szundy 1978, Duraes et al. 1995, Waes 1995) suggest that the size of the growing area did not significantly influence the seed germination ability, but had an effect on the grain yield. Twelve maize hybrids and their reciprocals were grown at three stand densities (51,000, 68,880, 94,390 plants/ha) at five locations (Keszthely, Gönc, Gyöngyös, Sopronhorpács, Martonvásár) for two years. After harvesting and drying, the seed were germinated according to the standard (MSZ 6354/3-82) in a Conviron climatic chamber for seven days at 25°C and 75 % RH. 1. The location caused a significant difference in the germination ability of the hybrid seeds. Averaged over the two years, samples from Gyöngyös had the poorest germination (89.1 %). 2. The size of the growing area had no significant effect on seed germination ability. 3. The year caused no significant difference in the germination ability of either the parental lines or of the hybrids. 4. In most cases the differences between the laboratory germination ability of seed of direct and reciprocal hybrids were statistically significant. In both years the same direct or reciprocal variant germinated better for each hybrid. 5. In both years the germination of the parental lines was compared with that of the direct and reciprocal hybrids. Averaged over the years, the parental lines had an average germinating ability of 90.3 %, while this figure was 95.3 % and 95.6 % for the direct and reciprocal hybrids. The degree of heterosis ranged from 102–114 %, being highest in Gyöngyös (114 %) and much the same (102–105 %) at the other four locations. The results showed that the climatic conditions at the growing sites affected seed germination ability, while, as found by other authors, the size of the growing area had no significant effect on this property. In both years there was atmospheric drought in Gyöngyös, causing a substantial reduction in the biological value of the seed. In several cases the cytoplasm had a significant effect on the germination ability of the direct and reciprocal hybrids.

75 27th ISTA Congress – Seed Symposium

98 - LASER IRRADIATION EFFECTS ON MAIZE SEED VIGOUR HERNÁNDEZ AGUILAR¹, C., CARBALLO, C.², A., ARTOLA², A. & MICHTCHENKO¹, A.

¹Instituto Politécnico Nacional, Sepi-Esime, Zacatenco. Unidad Profesional “Adolfo López Mateo”. Col. Lindavista. México D.F., México. CP 07738, [email protected] ²Colegio de Postgraduados. IREGEP. Programa de Semillas. Montecillo. Edo. de México. México. CP 56180

High seed quality is absolutely necessary in the context of modern crop production systems, and it is a specific requirement because affects crop establishment, growth and yield. Seed vigour is an important seed quality component related with a rapid and uniform seedling emergence and development. Considering that a chronic problem in the seed industry is the production of seeds with low seed vigour; therefore, the development of a technique to enhance it is very important. For this proposes, the effects of low intensity laser irradiation on Zea mays L. seed vigour was studied. A factorial of three intensities and six times of AsAIGa laser radiation, with an output power of 30 mW and wavelength of 660 nm, was applied in pre-sowing to seeds from a single cross hybrid. The laser seed irradiation significantly increased seedling emergence rate, seedling dry weight and emergence in soil. Due that laser irradiation originated positive and negative bio-stimulations in seed vigour, it was very important to estimate the specific combination of parameters. In this study, the optimum combination was an intensity of 20 mW/cm2 during one minute; this treatment originated highly significant (P ≤ 0.01) increases in seedling emergence rate (42.89%), in dry weight (63.24%) and emergence (48.07%) with relation to the control (without irradiation). From these results the laser irradiation seed enhancement technology deserves further investigation in maize.

99 - AN AUTOMATED SYSTEM FOR VIGOR TESTING THREE-DAY-OLD SOYBEAN SEEDLINGS HOFFMASTER1, A.L., FUJIMURA1, K., MCDONALD*2, M.B. & BENNETT2, M.A.

1Department of Computer and Information Science, The Ohio State University, 2015 Neil Ave., Columbus, Ohio 43210-1210 USA, [email protected]) 2Department of Horticulture and Crop Science, 2021 Coffey Rd., The Ohio State University, Columbus, Ohio 43210-1086 USA

An image processing computer application to automatically assess the vigor of three-day-old soybean (Glycine max [L.] Merrill.) seedlings was developed. The software operates on acquired digital images of soybean seed lots placed on a paper towel. Soybean seedlings were extracted away from the paper towel and converted into various digital representations. These representations were used to analyze the seedlings and segment them into normal and abnormal categories. The normal seedlings were further processed so that a one-pixel-wide summary structure of the shape of the seedling was produced. From this summary structure, the software classified the seedlings into six type categories based on their shape. Each normal seedling was processed to remove the cotyledon portion of the summary structure based on the type category it fell into. The remaining summary structure, with the cotyledon removed, was then used to compute the length of each seedling in pixels. From these length measurements, speed of growth and uniformity of growth values were computed. These two values were normalized and combined into a zero to 1,000 vigor index for the seed lot. Combined with the post-processing corrective features, this computer software was able to achieve highly accurate and standardized measurements of each soybean seedling, providing an alternative to the current method of manually measuring soybean seedlings for speed and uniformity of growth when performing a vigor test.

76 Budapest, Hungary, May 17th – 19th, 2004

100 - EFFECTS OF FERTILISATION DATES AND NITROGEN FORMS ON SEED WEIGHT AND GERMINATION CAPABILITY OF MAIZE INBRED LINES HOJKA1, ZDRAVKO, BOGDANOVIĆ2, DARINKA, MIRIĆ1, MLADEN, SREDOJEVIĆ1, SLOBODANKA DRAGIĆEVIĆ1, VESNA & CVIJANOVIĆ1, GORICA

1Maize Research Institute, Zemun Polje, Belgrade-Zemun, Serbia and Montenegro 2Faculty of Agriculture, University of Novi Sad, Novi Sad, Serbia and Montenegro

The present study was aimed at the determination of effects of different fertilisation dates and nitrogen forms on seed weight and germination capability of maize inbred lines. Two maize inbred lines (PL 142 and PL 167) were fertilised with the following fertilisation -1 -1 combinations: (Nmin) - PK: 60 kg P2O5 ha and 60 kg K2O ha applied in autumn (const.) + fertilisation on the basis of the N-min method, with the addition up to 100 kgNha-1 (in -1 spring); Nautumn - PK (const.) + N - 100 kgha (applied in autumn); (Nspring) - PK (const.) + N -1 -1 - 100 kgha (applied in spring); (N1/2) - PK (const.) + N - 100 kgha (1/2 applied in spring -1 and the other 1/2 in autumn); (N1/3) - PK (const.) + N - 100 kgha (1/3 each applied in autumn, spring and side dressing), in the forms of amide, ammonium-nitrate and ammonia, as well as, control with no fertilisers. The 4-replicate trial was set up according to the randomised complete-block design in the experimental filed of the Maize Research Institute, Zemun Polje, in the vicinity of Belgrade. Seed quality was analysed after harvest of maize inbred lines in accordance with ISTA Rules. A 1000-seed weight and germination capability significantly varied in studied inbred lines in dependence on fertilisation dates and nitrogen forms. The highest seed germination capability of maize inbred lines was obtained with the nitrogen fertiliser applied in autumn. Furthermore, germination capability was significantly higher with the application of ammonium-nitrate in the form of KAN fertiliser than with the application of other nitrogen forms. The general conclusion is that the N1/3 variant with KAN as well as Nautumn with any other nitrogen form favours the inbred PL 142 (during humid years), while Nmin and Nautumn variants are suitable for the inbred PL 167 regardless of certain differences in humidity over years.

101 - EVALUATION OF BARLEY SEED VIGOUR AND ITS RELATIONSHIP WITH FUNGI INFECTION HRSTKOVÁ, P. & CHLOUPEK, O.

Mendel University of Agriculture and Forestry Brno, Czech Republic

The quality of spring malting barley caryopses (Hordeum vulgare L.) was examined using several parameters. Thirty six seed lots obtained from variety trials harvested in 2002 were evaluated. Vigour estimated as germination under stress conditions, i.e. low temperature and water stress, showed large differences among seed lots, which ranged from 20,5 to 100% of germinated seeds. However the influence of location (i.e. environmental conditions) on vigour was much higher than that of cultivar, statistically significant differences among cultivars were found. Another criterion was a fungi infection characterised by the presence and quantity of ergosterol. Significant negative correlation between vigour and ergosterol content suggests that the ergosterol analysis may contributed to the description of seed quality.

Acknowledgements: This research was supported by the Grant Agency of the Czech Republic (No. 521/02/153). We are grateful to Prof. Dr.Dr.h.c.W. Opitz von Boberfeld for ergosterol analyses.

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102 - AN ALTERNATIVE VIGOR TEST FOR COLD TEST IN MAIZE H. ILBI1, H., KAVAK1, S., DUMAN1, I., ESER2, B., ILKER3, E. & GOKCOL3, A.

1Ege University, Fac. of Agriculture, Dept. of Horticulture, Bornova, Izmir, TURKEY, [email protected] 2Ege University, Seed Technolgy Center, Bornova, Izmir, TURKEY 3Ege University, Fac. of Agriculture, Dept. of Field Crops, Bornova, Izmir, TURKEY

This research was conducted to investigate an alternative test for cold test and field emergence test to evaluate maize seed vigor. Standard germination test, cool germination tests at different temperatures (12, 15 and 18°C), cold test and field emergence test were performed in twelve maize seed lots. The cool germination at 15°C had the highest correlation with field emergence and percentage of the high vigorous seedling in cold (r=0,881 and r=0,908, respectively). Standard germination test was found to have less correlation with field emergence than the cool germination test at 15°C. The cool germination test can be an alternative test for cold test to evaluate vigour differences between lots.

103 - PROMISING VIGOR TESTS FOR AGED ONION SEEDS ILBI, HULYA & ESER, BENIAN

Ege University, Faculty of Agriculture, Department of Horticulture, Bornova, Izmir, Turkey [email protected], [email protected]

Twenty-eight seed lots of two-onion cultivar (Diana F1 and Aki) were created by aging seeds naturally to investigate the efficiency of the vigour tests related to aging. All seed lots were subjected to the following tests: germination percentage and abnormal seedling rate in standard germination test, cool germination percentage and abnormal seedling rate, electrical conductivity (E.C.), sugar and amino acid leakage from seed. As a consequence of seed deterioration by ageing, the standard germination percentage declined in low vigour lots. E.C., sugar and amino acid contents in seed leachate increased related to loss of seed vigour. The sugar leakage from seed lots was found to be highly correlated with performance in stress condition e.g. cool germination test (r=-0,927), hence sugar leakage test appears to be suitable for evaluating seed quality in onion seeds and could be further developed as a rapid vigour test.

104 - INFLUENCE OF ACCELERATED AGEING TECHNIQUE ON SEED QUALITY EVALUATION JAGADISH, G.V.

Indo American Hybrid Seeds(I) Pvt.Ltd., Bangalore, India

In high, medium and low vigour seeds of tomato, capsicum, onion, French bean and Radish; Physiological and bio-chemical parameters were drastically reduced due to the degree of accelerated ageing (AA). The germination, seedling vigour and emergence were affected in seed lots subjected to harsh AA of 98% RH and 42°C for 48 hrs (72 hrs in Radish). The growth and yield were also reduced significantly compared to mild AA of 80% RH and 42°C for 48 hrs (72hrs in Radish) and non-aged seed lots. The correlation between physiological and biochemical attributes amply revealed that the seedling growth rate parameters were positively and significantly correlated to germination and field emergence potential. Seed quality was found to differ significantly among the high, medium and low vigour levels. Germination, seedling growth rate, seedling vigour, hypocotyls length, first counts germination And field emergence were reduced with the decrease in vigour levels. In the field conditions also the manifestation of vigour levels was extended; there was a reduction in the growth rate, fruit Weight and number of fruits per plant and fruit yield.

78 Budapest, Hungary, May 17th – 19th, 2004

105 - THE CONDUCTIVITY TEST FOR RICE (ORYZA SATIVA) MIN-HEE JEONG & SANG-KEUN LEE

Central Seed Testing Laboratory, National Agricultural Products Quality Management Services (NAQS), 560, 3-Ga, Dangsan-Dong, Youngdeungpo-Gu, Seoul, Republic of Korea

This study has carried out by the conductivity test, the most commonly used and evaluated as reproducibility method which included ISTA rules, to predict field emergence more exactly and to reduce the period for germination test at the certificate stage of the rice in our country. The purpose of this experiment is to set up measurement conditions and evaluation standard of the rice. To examine the relationship between conductivity and standard germination values it was used samples of germination group like below 85%, 85-90% and above 90%. The hulled rice was used to be leak effectively. Initial seed moisture contents of all samples were between 12-14%. The conductivity meter was chosen with a dip cell having a cell constant of 0.1 and a range of 0.5-200 µscm-1 because of small quantity of leakage in comparison with Fabaceae. Results of the conductivity for the hulled rice were greater 200 seeds than 400 seeds in 150 ml soak water at 25°C, but did not differ in 200ml soak water at 20°C. The leakage time of 20 hrs was significantly increased conductivity value in 20 hrs and 24 hrs. According to the conditions of results, the conductivity value of the rice was a significant correlation with standard germination rate.

106 - DETERMINATION OF SOYBEAN SEED OSMOTIC POTENTIAL PRIOR TO GERMINATION UNDER STRESS CONDITIONS KHAJEH-HOSSEINI1, MOHAMMAD, POWELL1, ALISON A. & BINGHAM2, IAN

1Department of Agriculture & Forestry, University of Aberdeen, Hilton Campus, Hilton Place, Aberdeen, AB24 4FA, Scotland, U.K 2SAC, Department of Agronomy, Craibstone Estate, Aberdeen, AB21 9YA, Scotland, U.K

Measurement of the osmotic potential of seeds by thermocouple psychrometry is not suitable for use with seeds that have been subjected to stress during germination using PEG and NaCl, due to errors arise from contamination of cell sap by PEG and NaCl in the apoplastic water. The osmotic potential of soybean seeds was therefore assessed by a modified form of a moisture release (MR) curve. Soybean seeds (cv. Williams) were imbibed either in deionised water or 421 mM (-1.9MPa) NaCl solution then dehydrated to a constant relative water content (RWC) by placing them in PEG solutions at a range of water potentials from - 0.5 to -6.5 MPa. Seeds were also imbibed directly in a range of PEG (-0.5 to -6.5 MPa) solutions until they reached an equilibrium moisture content (MC) and the seed RWC calculated on the basis of a MC of 55% at full imbibition. The inverse of external water potential (1/ψ) at MC equilibrium was then plotted against the seed RWC, that is, as a MR curve which was used to determine seed osmotic potentials. At lower external water potentials of PEG solutions the seed osmotic potential was lower (i.e. more negative) in all cases. A lower osmotic potential at full turgor (RWC=100%) (-2.15 MPa) was observed for the seed pre-treated with NaCl compared with seed placed directly in PEG solutions (-1.79 MPa). MR curves give tissue-average values for osmotic potential, while the MC for the components of soybean at full imbibition are different. Therefore, the applicability of the MR technique may depend on the seed structure and the nature of seed water relations in different species particularly in stress conditions.

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107 - EFFECT OF TEMPERATURE AND STORAGE ON SEED VIABILITY OF VALERIAN (VALERIANA SISYMBRIIFOLIA) KHAYYAM NEKOUEI, MOJTABA & BOARDBAR, ZARRINTAJ

Department of Biotechnology, Isfahan Research center of Agriculture and Natural Resources, Iran, Isfahan, [email protected]

Valeriana sisymbriifolia is one of six species of valerian genus, which are distributed in north and center of Iran. Valerian is used as a relaxation of Central Nervous System (CNS), antispasm activity.The relaxant effective of moderate relaxation is from valepoteriates and sequie terpens. Because of problem for seed germination of Valeriana sisymbriifolia is not prevalent. Therefore producing high quality of seed and optimizing of storage condition are very important in this process. In this study the effect of temperature and length of seed storage on two accession of valerian were investigated. Collections of seed were conducted from Khoonsar and Feridonshahr in Isfahan province while at least 50% of mature seed papus were emergence and also the form of inflorescence was villous. The seeds were then sterilized and packaged in a three layer aluminum envelop and were used in two separate experiment under (-10°C, +10°C, +25°C) and (-5°C, +5°C, +25°C) conditions respectively. Germination test was checked during one year in first, third, sixth, ninth and finally twelfth month. The data were analyzed based on random experimental design as a factorial. The results showed that there was no significant difference for temperature storage, although the recommended temperature is between 0°C to 10°C while less than 0°C and more than 25C may effect on seed quality. For the Khoonsar accession, seed storage was significant after sixth month. Viability deterioration after ninth month occurred but there was no significant after ninth and twelfth month (After one year germination percentage decreased from 90% to 68%). For the Fereidonshahr accession also the same results (the germination percentage decreased from 95% to 75% after on year) were obtained

108 - GERMINABILITY OF TREATED SOYBEAN SEED AS AFFECTED BY OXYGEN DIFFUSION RATE UNDER THE DIFFERENT SOIL MOISTURE CONTENT KIM1, SEOK H. & TAYLOR2, ALAN G.

1Department of Agronomy, College of Agriculture and Life Sciences, Gyeongsang National University, Chinju 660-701, KR-Republic of Korea 2 Department of Horticultural Sciences, NYSAES, Cornell University, Geneva, NY 14456, US-United States

Seed germination has been considered largely in relation to seed viability under the conditions of favorable moisture and oxygen supply in soil. We conducted experiments with two seed lot and two Captan treatments including 1) Control, 2) Control + Captan, 3) one week-aged, and 4) one week-aged + Captan. The oxygen diffusion rate (ODR) was assessed under the different soil moisture contents, 17, 18, 19, 20 and 20.5% in the sandy loam soil. The aim of the present study was to determine the ODR that is characterized the optimum oxygen conditions in the soil and plays an important role in the emergence depending on vigour of soybean seeds. We discovered a critical point of soil moisture content that is lower than 20.5%, because the seed emergence was sharply decreased coupling to ODR decrease under 36.4 µg/m2.sec during conducting experiment. As 19 ~ 20 % of soil moisture content levels, the percent emergence and number of normal seedling were greater at the seeds treated with Captan compared to control. However, no difference was observed between with or without Captan treatment at different aged seeds. In the results, the value of 17% soil moisture content might be considered as the optimum condition for soybean emergence, when ODR was reached 101.8 µg/m2.sec. The finding could provide a new insight on the germination of soybean seeds about the relationship between the seed vigour and soil moisture contents.

80 Budapest, Hungary, May 17th – 19th, 2004

109 - PROLINE: USE AS AN INDICATOR OF TEMPERATURE STRESS IN SEEDS MACHADO NETO1, NELSON BARBOSA, CASTILHO CUSTÓDIO1, CECI, GATTI3, ANA BEATRIZ, PRIOLLI3, MARIA REGINA & MENDES CARDOSO2, VICTOR JOSÉ

1Universidade do Oeste Paulista - Curso de Agronomia- UNOESTE - Rodovia Raposo Tavares Km572, Presidente Prudente SP 19067-175, [email protected]; [email protected] 2, 3Universidade Estadual Paulista - Campus de Rio Claro - Av.24A, 1515 Rio Claro SP

Stress due to temperature could lead to several metabolic alterations during the hole plant cycle including germination. To verify the effect of high or low temperatures, constant or in alternate cycles, 10 populations of beans seeds, 8 cultivars and two landraces, were submitted to distinct temperature cycles of 8, 18 or 37ºC during 96 h, or during 48h at 18ºC and after at 8 or 37ºC during more 48h. Among the cultivars, ‘IAC-Carioca-80SH’ and ‘Rosinha G2’, were multiplied in replicates in two distinct fields, one in Rio Claro (SP) and other in Presidente Prudente(SP). Dehulled seeds were grind in 3% sulphosalicylic acid. The resultant extract was double filtered and reacted with acid ninhidrin, being heated at 100ºC during 1h and immediately refrigerated in a cold bath. Proline was quantified at 520nm in a spectrophotometer. There were responses to different environments for each population, and there were also different answers between production fields, e.g., seeds produced in Presidente Prudente did not exhibited variations to alternate temperature during germination. Proline could be used as an indicator of stress temperature.

110 - ASSESSMENT OF SEED VIGOUR AND STORABILITY OF HYBRID AND PARENTAL LINES OF SORGHUM MANOHAR REDDY, N., ANKAIAH, R. & MURALIMOHAN REDDY, B.

National Seed Project, ANG Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, India, [email protected]

Seeds of SPH-468, SPH-504, CSH-9, CSH-6 and CSH-5 sorghum hybrids and their parental lines viz., MS 2077 A, CS 3541, 296A having above 80% germination and 8.5% initial seed moisture were subjected to accelerated ageing by keeping the seed in environmental chamber at 40±1°C and 85±2% RH for 4, 8, 12, 16 and 20 days. The germination percentage, seedling length and vigour index gradually decreased with the increase in period of accelerated ageing as a result of seed deterioration in hybrids as well as parents in all genotypes. Such decline in germination, seedling length and vigour index was high in 296 A, CSH-5, SPH-504 and CSH-6 and these were considered as poor stores, SPH-468, CSH-9, CS- 3541 were considered as good storers due to higher values of germination and vigour.

111 - IMBIBITION DAMAGE IN DRIED BEAN SEEDS MARTÍN, ISAURA, DE LA ROSA, LUCÍA & DE LA CUADRA, CELIA

Centro de Recursos Fitogenéticos (CRF). INIA. Apartado 1045, 28800 Alcalá de Henares, Madrid, Spain.

The accurate estimation of seed viability is a crucial aspect for controlling genetic erosion in genebanks and therefore germination protocols should be set up carefully in each species. At the CRF genebank the seed viability of every accession is estimated before storage and every ten years, through germination tests conducted under ISTA rules, with the necessary adaptations in each case. Incidence of imbibition damage has been detected in the routine CRF germination tests of some large-seeded legumes, such as beans. Imbibition injury is greater in very dry seeds and seed hydration pre-treatments are required when samples are stored at low moisture contents. In this work the effect of different seed moisture contents on seed imbibition damage was studied in six bean Spanish landraces. Three of them (V1,V2,V3) had white seed coats and the other three (V4,V5,V6) were dark coloured. Seeds of each variety were progressively desiccated, first in dehydration chamber (20ºC, 20% R.H.) and then in silica gel. Hydration treatments at 100% R.H. were also performed in 81 27th ISTA Congress – Seed Symposium order to obtain seeds with high moisture levels. Ten moisture contents were established, ranging from 5% to 25% m.c, and then seed viability and vigour were evaluated through germination and electric conductivity tests. The electric conductivity measures showed a narrow relation with the seed moisture content. A clear conductivity increase was observed when seed moisture decreased, except for one dark-coloured variety, V4, in which conductivity values remained constant. The strongest desiccation treatments induced some degree of hardseededness, specially in the variety, V5, this fact modifying the conductivity pattern. As the conductivity indicates the cellular membrane stability, these results demonstrate the inverse relation between the seed moisture degree and the injuries provoked by a rapid imbibition in all landraces, except for V4, that showed no susceptibility to this type of damages. Same conclusions were obtained from germination tests although in that case the differences obtained were less pronounced. In other studies, white seeds were generally more susceptible to imbibition damages than dark seeds, but in our case the dark seed varieties V5 and V6 showed similar injuries than those with white seeds.

112 - EMERGENCE OF MAIZE IN COLD SOIL MARTON1, L.C., KIZMUS1, L., HEGYI1, ZS. & ZÁBORSZKY2, S.

1Agricultural Research Institute of the Hungarian Academy of Sciences, Martonvásár 2Georgikon Faculty, Veszprém University, Keszthely

In Hungary the maximum yields are obtained from maize sown between 10–20 April. It is generally said that sowing can be begun when the soil temperature at the sowing depth no longer drops below 10°C. However, even when sowing is carried out at the optimum time, it may be followed by relatively long periods of cold weather leading to soil temperatures of less than 10°C. Investigations were thus made on the effect of cold spells of various length on the emergence of maize. The effect of sowing date (April, 3, 10 and 17), seed dressing agent (Captan, Carboxin + TMTD and untreated control) and soil (heat-sterilised maize soil, sand, maize soil infected with Fusarium) on emergence was studied in a pot experiment, averaged over 5 maize hybrids. The results indicated that a long period of cold did not in itself reduce the emergence percentage. The pathogen, however, killed almost 100% of the undressed seeds. The dressing agents provided considerable protection, since there was 87% emergence even in the artificially infected treatment. A complete stand can thus be obtained even from very early sowing at the beginning of April if seed dressing is carried out efficiently. The number of days from sowing to emergence depended decisively on the sowing date. At the earliest sowing date the emergence time was twice that observed at the optimum sowing date. The time to emergence was only shortened by about half a day by dressing, averaged over the treatments. No significant difference was observed between the different sowing dates for the accumulated heat sum required for emergence. The emergence time is thus determined to the greatest extent by the temperature. As the result of the earliest sowing date the stand obtained with dressed seeds was more vigorous and their dry shoot mass was greater, when measured on the same date, than those emerging from later sown seed. The dry mass of plants emerging from undressed seeds sown in infected soil at the earliest sowing date was only 12% of that in the dressed treatment. Even at the 2nd and 3rd sowing dates plants originating from undressed seeds were only able to develop 70–72% of the dry shoot mass of plants from dressed seeds in infected soil. Soil-borne pathogens are thus capable not only of substantially reducing the emergence percentage, but also of retarding the growth and development of the emerged plants.

82 Budapest, Hungary, May 17th – 19th, 2004

113 - EFFECT OF FUNGICIDES ON STORABILITY OF RICE HYBRIDS AND THEIR PARENTAL LINES MEENA KUMARI, K.V.S., SAVITRI, H. & MURALIMOHAN, REDDY B.

National Seed Project, ANG Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, India, [email protected]

Hybrid rice seeds of DRRH-1 (IR 58025A x IR 40750R), APHR-2 (IR 62829A x MTU 9992), ADTRH-1 (IR 58025A x IR 66R) and their parents having an initial germination of 85-90%, 9-11% seed moisture and 4-12% total fungal colonies (Curvularia lunata, Drechslera oryzae and Fusarium moniliforme) were utilized for storability studies during 2000. The seed samples treated with thiram and captan @ 0.25% along with untreated seed were stored in two containers viz., cloth bag and polythene bag (700 gauge) under ambient storage conditions at National Seed Project, Rajendranagar, Hyderabad, India. The observations on germination percentage, seed moisture content, seedling vigour and total fungal colonies at bimonthly intervals upto 18 months were recorded. Irrespective of seed treatments and containers used in the presentstudy, restorer lines (male parents) recorded higher germination (80%), seedling vigour (1625) with lesser fungal colonies (5-25%) during the entire storage period. Among the genotypes, IR 66R had a maximum storability of 16 months, while DRRH-1 had 10 months with a germination percentage (80%) and fungal colonies (10%), respectively. The present investigation revealed that hybrids had less storability (10-12 months) over their female parents (12 months) and restorer lines (16 months). Irrespective of genotypes and treatments employed, seeds stored in polythene bag exhibited above 80% germination upto 14 months as against 8 months in cloth bag. Seed treated with thiram and captan @ 0.25% had profound influence on genotypes particularly in polythene bag in extending the seed longevity as compared to control in both the containers. The improvement in storability of treated seed in polythene bag was due to less fungal colonies. Captan treated seed stored in polythene bag was very effective in improving the shelf life of restorer lines (17 months) and female parents (14 months). Thus, the precious seed of hybrids and parental material can be stored safely for 1 to 1 ½ years with captan treatment and stored in polythene bags, respectively.

114 - ACCELERATED AGEING TEST FOR WHEAT (TRITICUM AESTIVUM): REPRODUCIBILITY OF TWO AGEING METHODS MERIAUX1, BENOIT, LADONNE2, FABIENNE & FOUGEREUX1, JEAN-ALBERT

1FNAMS, Le Verger, 49800 Brain sur l’Authion – France 2LABOSEM, Le Verger, 49800 Brain sur l’Authion – France [email protected]

In order to help French seeds companies for storage management of wheat seeds, we had to develop a test for a routine use. Accelerated ageing has been shown to be a predicting test for viability after storage but different methods can be used. The aim of the present study was to compare the reproducibility of two ageing methods. Seeds were aged at 45 C during 3 days upon water (~ 100% Relative Humidity) or during 7 days upon a saturated solution of sodium chloride (~ 75% Relative Humidity). After ageing, percentage of normal seedlings was determined with a germination test. Each method was applied 4 times on 4 seed lots without or with fungicide seed treatment. At 100% RH, differences of germination values between rounds were very important (up to 45%) and greater than tolerances for ageing test in ISTA' Rules. At 75% RH, differences between rounds were reduced and always lower than the ISTA’s tolerances for germination test. Upon water, fungi development during ageing and / or germination test was observed. But the poor reproducibility of ageing method upon water was probably not only due to fungi if we consider that reproducibility is not improved by seed treatment. These results indicate that accelerated ageing test upon saturated solution of sodium chloride is more reproducible than accelerated ageing upon water.

83 27th ISTA Congress – Seed Symposium

115 - DETERMINATION OF STORAGE POTENTIAL OF WHEAT SEEDS: RELEVANCE OF AN ACCELERATED AGEING TEST USING NACL MERIAUX1, BENOIT, FOUGEREUX1, JEAN-ALBERT & LADONNE2, FABIENNE

1FNAMS, Le Verger, 49800 Brain sur l’Authion – France 2LABOSEM, Le Verger, 49800 Brain sur l’Authion – France [email protected]

Temperature, moisture content and storage potential of seeds affect their viability during storage. We have developed an accelerated ageing test using a saturated solution of sodium chloride (75% RH) to determine the storage potential of wheat seed lots. The aim of the study was to determine the relevance of this test applied to commercial wheat seed lots. From 2000 to 2002, 46 wheat seed lots were collected at harvest from seeds companies. For each lot, a sample of seeds was untreated and another was treated with fungicide. In november, accelerated ageing test (7 days at 45°C upon NaCl saturated solution) was applied to all the seed lots. At the same time, unaged seeds were stored in ambient condition or in controlled unfavourable condition (30°C and 15.5% seed moisture content). Seed quality was evaluated during storage a germination test. In ambient condition, germination kept a high level after one year. In unfavourable storage condition, ageing is faster and, after 75 days ageing we observed large disparities between seed lots. It appeared that seed treatment slightly increased ageing. Germination after storage in unfavourable condition and results of accelerated ageing test were close correlated (R² from 0.588 to 0.845) for both untreated and treated seeds. For untreated seeds, correlations were slightly lower when fungi development was observed during storage. Our results show that the accelerated ageing test using saturated solution of sodium chloride is a relevant test to determine the storage potential of wheat seed lots.

116 - PREDICTION OF GERMINATION IN BRASSICA SPP USING THE BULK CONDUCTIVITY TEST MIRDAD1, ZOHAIR, POWELL2 ALISON A. & MATTHEWS2, STAN

1Department of Arid Land Agriculture, King Abdul Aziz University, P.O.Box 80208, Jeddah, 21589 Saudi Arabia, [email protected] 2Department of Agriculture and Forestry, University of Aberdeen, Scotland AB24 5UA, UK

This research aimed to determine whether measurements of solute leakage from seeds of cauliflower and cabbage by the bulk electrical conductivity test could be used to predict germination. Two approaches were used. Firstly, samples differing in germination were created by combining different proportions of living and dead seed within the test sample from 100% living to 10% living. This resulted in a clear correlation (R2=0.99) between conductivity and actual germination. The second approach was to produce samples having different germination as a result of ageing under different conditions. Seeds were therefore aged at two moisture contents (15, 20%), for 0-120 h at 45°C. Germination decreased and conductivity increased with increased ageing time in both species. Where there was a wide range in the germination of aged samples (1- 100%), there was again a correlation between germination and leachate conductivity (R2 > 0.92). When germination was high (90-100%) conductivity was closely correlated with the germination of cauliflower (R2 > 0.70) although less so for cabbage (R2 > 0.46). Conductivity and germination values from both ageing treatments combined to give a single correlation of 0.92 (normal germination) and 0.87 (total germination) for cauliflower and 0.80 (normal germination) and 0.75 (total germination) for cabbage. Thus the leakage / germination relationship was apparently independent of the rate of ageing as influenced by seed moisture content.

84 Budapest, Hungary, May 17th – 19th, 2004

117 - MAIZE SEEDS SHAPE AND POSITION IN THE EAR: EFFECTS ON THE QUALITY EVALUATED BY IMAGE ANALYSIS MONDO VITOR, HENRIQUE VAZ & CICERO, SILVIO MOURE

São Paulo University, Av. Pádua Dias, 11, C.P. 09, 13418-900 Piracicaba – SP, Brazil, [email protected]

The possibility of using the image analysis technique for assessing the quality of seeds is very promissing; it is a precision method that enables one to examine, in detail, the damaged or altered region, its location and extent. As it is a non-destructive method, the seeds being analyzed can be submitted to physiologic tests and thus, allow the relations of cause and effect to be established. Thus, the objective of this research was to study the effects of the positions of maize seeds on ears, on the quality, using the image analysis technique. For this purpose, ears of the cultivar DKB 212 were shelled in such a way as to be able to isolate the seeds from the proximal, intermediate and distal positions. The seeds coming from the positions referred to were physically characterized and assessed with regard to vigor and sanitary condition. Simultaneously, the seeds were radiographed, photographed (ventral and dorsal faces) and destined for germination test; the normal, abnormal seedlings and dead seeds were also photographed. All the images obtained were made available on the computer in such a way that they could be examined simultaneously and thus make a complete diagnosis for each seed. Results indicate that the seeds from the proximal and intermediate positions show a similar quality and are superior to those from the distal position.

118 - EVALUATION OF VIABILITY OF OIL PALM (ELAEIS GUINEENSIS. JACQ.) SEEDS BY TETRAZOLIUM TEST AND COMPARISON WITH GERMINATION AND IN VITRO CULTURE RESULTS MURUGESAN, P., VANANGAMUDI, K. & UMARANI, R.

Department of Seed Science and Technology, Tamil Nadu Agricultural University, Coimbatore – 641 003, Tamil Nadu., [email protected]

Oil palm (Elaeis guineensis Jacq.) requires about two months of high temperature (40°C) treatment to overcome its dormancy and 30 days for completion of germination. To evaluate its viability within a short period of time different concentrations of Triphenyl Tetrazolium Chloride (TTC) test was tried to standardize the duration which gives satisfactory embryo staining in order to distinguish viable seed with that of non viable. The excised embryo from Dura (255D) was tested with TTC and compared with results obtained from in vitro culture of embryo and standard germination test. The TTC concentrations of 0.5 to 0.75% was appeared to gave satisfactory staining when treated embryos were kept under incubation at 40°C for four hours which was in close agreement with other tests.

119 - STUDY OF HEAT TOLERANCE DURING GERMINATION IN GRAIN LEGUMES NEMESKÉRI, ESZTER

Agrona Ltd. Debrecen, Domokos M. Kert 72, H-4002 Debrecen Hungary

In this work the effect of high germination temperature has been investigated on seed quality of soybean varieties and bean breeding lines grown under irrigated and nonirrigated, dry conditions. The seeds were germinated at 24°C and at high temperature (28°C) in automatic thermostat. After four and eight days we expressed the degree of drought tolerance by the rate of abnormal and diseased seedlings. The bean seeds were very sensitive to high germination temperature, which doubled the rate of diseased seedlings compared with controls. The high temperature significantly increased the number of abnormal seedlings, which come from seeds of both species grown under nonirrigated, dry 85 27th ISTA Congress – Seed Symposium conditions. At the early stage of germination the selection to improve heat tolerance of beans based on abnormal seeds could be achieved at high temperature (28°C). The difference in drought tolerance among soybean varieties can not be expressed by the rate of increasing abnormal seedlings. At 28°C germination temperature the root development of bean seedlings was hampered; the average primary root length of bean varieties was 31.35 percentage of the control (24°C). The decrease was small (70.43%) in soybeans, the development of root seedling was hardly hampered under 28°C temperature. It appears that large-seeded bean lines with stronger roots were less susceptible to high temperature than the small-seeded lines

120 - INVESTIGATION ON GERMINATION ABILITY OF SOLIDAGO SPECIES (SOLIDAGO VIRGA-AUREA L., SOLIDAGO CANADENSIS L., SOLIDAGO GIGANTEA AIT.) NOVÁK I.1, DRAHOS ZS.1, TÓTH E.2 & JANIK, G.3

1 BKÁ University, Faculty of Horticulture, Department of Medicinal and Aromatic Plants, Budapest 2 Ökoherba Ltd., Herencsény 3 Pannon Flóra Ltd., Budaörs

Solidago virga-aurea L. has been introduced into the production in the recent years, however, the germination biology of this species has hardly been investigated. The two adventive species (Solidago canadensis L. and Solidago gigantea Ait.) are expanding intesively in Hungary and hardly any scientific data has been published on their germination capacity yet. Therefore the aim of our study was to investigate the germination features of this three Solidago species. After one year of storage period at +4ºC, the germination capacity of S. virga-aurea decreased significantly (from 62.5% to 24%), while in case of S. gigantea an after-ripening process was observed, in harmony with former findings. The germination capacity of this species increased significantly from 35% to 73%. The germination rate of the achenes of S. canadensis has slightly increased: from 82.5% to 86% under the same storage circumstances. This high germination capacity and after-ripening process may contribute to the spread of the two adventive species in Hungary. The achenes of S. virga-aurea originated from 11 different places from Hungary germinated from 48% to 76%. The differences in the germination capacity remained accession dependent even if they were grown on the same experimental field, and the achenes were collected from the same circumstances.

121 - TETRAZOLIUM TEST FOR DETERMINING VIABILITY OF BRACHIARIA BRIZANTHA SEEDS NOVEMBRE1, ANA DIONISIA L.C., CAMMA1, HELENA M.C.P. & GOMES2, ROSANGELE B.R.

1USP/ESALQ/LPV, CP 09, 13418-900, Piracicaba/SP, Brazil, [email protected] 2SFFV-DDR/DFA/SP, Rua 13 de maio, 1558, 01327-2002 - São Paulo/SP., Brasil.

The objective of this research was to evaluate the viability of Brachiaria brizantha seeds by tetrazolium test. Eight seed lots preconditioned between paper during two, four and six hours at 35°C and 40°C or six and 16 hours at 30°C. Seeds were then bisected longitudinally through the embryo and immersed in a 0,075% tetrazolium chloride solution (staining at 40°C for two hours). After those preliminary tests, seeds imbibed for six hours at 30°C were staining during one, two and four hours at 35°C, 40°C and 45°C or six hours at 30°C. Seed water content (oven method, 105±3°C/24h) and germination (TP, 20-35°C, count at 7, 14 and 21 days) were determined. The tetrazolium test is an efficient option to estimate the viability of Brachiaria brizantha seeds. Consistent information resulted from the combination of preconditioning at 30°C for six hours (seed water content of 25%) followed by a two hours staining at 40°C. This represents a considerable reduction of time needed to perform the tetrazolium test for Brachiaria brizantha.

86 Budapest, Hungary, May 17th – 19th, 2004

122 - STORAGE STUDIES IN RICE HYBRID PATIL N.K.BIRADAR & SHEKHARGOUDA, M.

University of Agricultural Sciences, Dharwad -580 005.

The success of hybrid rice technology not only depends on the production of quality seeds in large quantity, but also on maintaining the seed viability and vigour till the next sowing season. Hence, the knowledge on seed storability of rice hybrids and their parents is very essential to avoid huge financial losses. The results of the present investigation reveal significant influence of genotypes, production location, seed treatments, packaging material and storage period on seed viability and vigour. Irrespective of other factors, the seed quality parameters decreased steadily with the progress in storage period. The seed produced at Dharwad location record significantly higher seed quality parameters compared to Sirsi. The seed viability and vigour varied among the genotypes throughout the storage period. The hybrid seed and the male parent were found to be better store, whereas maintainer line exhibited poor storability. The seeds stored in polythene bag were superior over cloth bag for all quality parameters during the storage. The untreated seeds lost viability and vigour at a faster rate during storage compared to Thiram treated seeds.

123 - COMPARISON OF THE THOUSAND KERNEL MASS AND GERMINATION ABILITY OF MAIZE (ZEA MAYS L.) LINES GROWN IN HUNGARY AND CHILE PINTÉR, J., HEGYI, Z., BERZY, T. & MARTON, L.C.

Agricultural Research Institute of the Hungarian Academy of Sciences, Martonvásár, Hungary

For almost ten years now the Martonvásár Institute has maintained a winter generation programme in Chile, in the southern hemisphere, where the climatic and ecological conditions are approximately the same as those in Hungary, in order to be able to raise two generations of maize a year. The UV-B level has been proved to be 25% higher in Chile on average, resulting in physiological changes, generally negative, in some inbred lines compared with those grown in Hungary. These include asynchronisation of flowering, barrenness, greater pollen mortality, etc. Since the situation is likely to worsen in Hungary, too, investigations were made to determine whether higher UV-B radiation had any negative effect on the seed quality of Martonvásár inbred lines. The data obtained would also allow a selection programme to be set up for the development of inbred lines with better tolerance of higher UV-B radiation. The seed yield of 52 inbred lines sown in Hungary in 1999 and in Chile in the winter generation programme in 1999/2000, all from the same source, was used for the measurement of thousand kernel mass and germination ability (according to the standard MSZ 6354/3-82). Germination took place in the phytotron, in a Conviron climatic chamber, for 7 days at 25°C with 75 % RH. The results obtained in the first year indicated that the germination ability of the Martonvásár lines was significantly better (92 %) than that of lines grown in Chile (90%). For some genotypes the Chilean data were far worse than the average. The determination of thousand kernel mass also gave better results in Martonvásár. The thousand kernel mass of the same lines was significantly higher in Martonvásár (267.6 g) than in Chile (246.1 g). The MQ values obtained from two-way analysis of variance indicated that the location had a greater effect on this parameter than the variety.

87 27th ISTA Congress – Seed Symposium

124 - THEORETICAL EFFECTS OF ALTERED BIOLOGICAL AND CHEMICAL PROPERTIES ON SALINITY TOLERANCE OF ACACIA SEEDS REHMAN, S1., KIM, J.K1., RHA, E.S2., ASHRAF, M.3 & HARRIS, P.J.C.4

1Faculty of Biological Resources Science, College of Agriculture, Chonbuk National University, Chonju 561-756, Republic of Korea. 2Life Resources Science, Plant Genetics & Breeding, Sunchon National University, Sunchon, 540-742, Republic of Korea. 3Botany Department, University of Agriculture, Faisalabad, Pakistan 4Biosciences, School of Science and Environment, Coventry University, Coventry, CV1 5FB, UK

Multiple regression equations (Rehman et al., 2000) have been developed to predict the salinity tolerance of Acacia seeds, expressed as the I50 (the concentration of NaCl required to reduce final germination to 50% of the control value in DW). Accurate predictions can be made using one or more chemical and biological seed parameters. In this study the theoretical effect of varying final germination percentage in distilled water, germination rate in distilled water (Rate), Ca2+ or K+ contents and their ratios, as independent factors or related factors, on the predicted salinity tolerance (I50) of Acacia species was investigated. Simulation of the effects of changing final germination, rate, calcium and potassium suggest the possibility of practical application of these results to modify the salinity tolerance of seeds. The predicted I50 increased with increasing final germination. Similarly, the higher the rate of germination, the higher the predicted salt tolerance of Acacia species. The Ca2+ content of seeds was found to be positively correlated with I50. Species with higher Ca2+ contents had a higher I50. This suggests that that I50 might be increased by increasing the Ca2+ contents of seeds by pretreatment with calcium salts or by supplying these to the mother plants.

125 - THE USE OF X-RAY FOR PREDICTION OF SEED VIABILITY IN CONES RØSOK BYE, HEIDI & MELAND EDVARDSEN, ØYVIND

The Norwegian Forest Seed Station, P.Box 118, 2301 Hamar, Norway, [email protected]

Introduction The Norwegian Forest Seed Station collects cones, produce and sell forest seeds to nurseries. Decisions on seed collections often have to be taken on a few days notice. X-raying is a tool for quickly estimation of seed viability in cone samples and therefore crucial for obtaining quality seed. However, the method can under certain conditions diverge from the standard germination test. The X-raying method is discussed in the light of a closer examination of a large number of cone samples and gained experience through many years of operation.

The X-ray analysis of germination capacity X-raying were performed on seeds from 105 Picea abies and Pinus sylvestris cone samples, collected in the period 1999–2003. A representative selection of 2 x 100 seeds from each cone sample, were X-rayed. After processing, seeds were classified on the film and germination capacity calculated, in accordance with a classification system and formula prepared by Milan Simak. A corresponding 21-day germination test, were used as comparison.

Results and experience gained Germination capacity between the two methods of analysis, varied with more than 10% in 29 samples with a maximum of 31%. Conditions that may have influenced the X-raying precision in these samples, can be explained both with technical and biological as well as human related factors, such as: • Use of coarse-grained film, old processing liquid and film frames not suitable for X- ray penetration • Large proportions of empty seeds in samples and the statistical model it self • Physiological immature seeds and seed damages invisible to X-rays • The experience of the laboratory assistant

88 Budapest, Hungary, May 17th – 19th, 2004

126 - THE EFFECT OF STORAGE ON GERMINATION OF ALFALFA (MEDICAGO SATIVA L.) RUTAR1, ROMANA & STJEPANOVIĆ2, MIRKO

1Agricultural Insitute of Slovenia, Hacquetova 17, 1000 Ljubljana, Slovenia, e-mail: [email protected] 2Faculty of Agriculture, 31000 Osijek, Croatia

The germination is the parameter of the seed quality and the storage conditions has the influence on it. Three different varieties of alfalfa were checked for germination capacity during thirteen years. The seed samples of three varieties: Osječka 10 = OS-10, Osječanka 88 = OS-88 and Slavonka were stored at +4°C and 70% relative moisture for thirteen years. Each sample was checked for germination capacity each year during the whole period of storage. The germination was determined according to the ISTA method: constant temperature of 20°C with the prechilling. Thirteen-year seed storage has shown that the percentage of normal seedlings was the lowest in the first year: 53% at Slavonka, 59% at OS-10 and 74% at OS- 88. The highest value was reached in two or three years: 79% at Slavonka, 82% at Os-10 and 85% at Os-88. Afterwards a continuous fall was perceived and after thirteen years the results are nearly similar as at the beginning: 51% at Slavonka, 59% at OS-10 and 75 % at OS-88. The percentage of dead seeds was the lowest in the first year: 1% at OS-10, 2 % at OS-88 and 5% at Slavonka, increased during the years and after thirteen years the results are: 37% at Slavonka, 26% at OS-10 and 14% at OS-88. The % of hard seeds was the highest in the first year: 17% at OS-88, 28% at Slavonka and 30% at OS-10. After two years only one third of hard seeds were left and than they were reduced to their half next year, in the following years no essential changes were noticed. The results after 13 years are: 3% of hard seeds were found at OS-10 and Os-88 and 5% at Slavonka.

127 - DRYING METHODS EFFECTS ON GERMINATION AND DORMANCY OF COMMON VETCH (VICIA SATIVA) HARVESTED AT DIFFERENT MATURITY STAGES NEZAR H. SAMARAH

Department of Crop Production, Jordan University of Science and Technology, Irbid P.O. Box 3030, 22110, Jordan, [email protected].

Common vetch seeds acquire the germination capability at specific developmental stage. Seed maturity and drying methods may affect seed desiccation tolerance, and seed germination and dormancy. Field experiment was conducted to study the effect of pod maturity and drying methods on seed germination and dormancy of common vetch. Common vetch plants were harvested at five pod developmental stages: beginning seed pod (BS), full-seed pod (FS), greenish-yellow pod (GY), yellow pod (Y), or brown pod (B) stage. Seeds of the harvested plants were dried at ambient condition (28 ± 2) in three methods: seeds dried without pods, seeds dried within pods detached from plants, or seeds dried within pods attached to the plants. Seed moisture content was recorded during the drying treatments. Seed dry weight was measured before and after applying the drying treatments. Standard germination test and speed of germination (as estimated by germination rate index) were measured for harvested seeds. Seeds harvested at BS and FS stage and dried without pods had significantly lower germination (< 55%) and GRI, and higher dormancy than seeds harvested at GY, Y, and B stages, indicating that seeds dried without pods acquired the maximum germination capability when seeds were harvested at or after GY stage (slightly prior to seed physiological maturity). Seeds drying within pods detached/or attached with plants improved the percentage of seed germination (> 92%), GRI, and reduced the percentage of dormant and dead seeds and abnormal seedlings when seeds were harvested at the BS and FS pod stages, indicating that “after ripening” of seed within pods reduced seed dormancy and improved seed desiccation tolerance, germination, and GRI. Seeds harvested at or after GY stage attained the maximum germination (> 95%) with no significant differences among the drying treatments. In conclusion, common vetch seeds dried without pod attained maximum germination when seeds were harvested at or after GY stage. Seed drying within pods detached/ or attached with plants reduced the rate of seed

89 27th ISTA Congress – Seed Symposium moisture loss, increased seed dry weight, reduced seed dormancy, resulting in improving seed germination and speed of germination when seeds were harvested at early developmental stages.

128 - ESTIMATION OF TOTAL PHENOLICS IN SOYBEAN (GLYCINE MAX L.) EXUDATES AND SEED QUALITY DURING ACCELERATED AGEING SIMIĆ1, A., SREDOJEVIĆ2, S., TODOROVIĆ1, M., ÐUKANOVIĆ2, L., DAMJANOVIĆ3, M. & SPASIĆ, M.1

1Faculty of Chemistry, University of Belgrade, Studentski trg 14-16, 11000 Belgrade, Serbia and Montenegro ([email protected]) 2Maize Research Institute, Zemun Polje, Slobodana Bajica1, 11080 Belgrade-Zemun, Serbia and Montenegro (E-mail: [email protected]) 3Galenika Fitofarmacija a.d., Batajnički drum bb, 11080 Belgrade-Zemun, Serbia and Montenegro (E- mail: [email protected])

Three soybean (Glycine max L.) cultivars were tested for response of viability to accelerated ageing. The freshly harvested seeds were high quality (the percentage of germination were higher than 95%). Samples aged at 44 ºC and nearly 100% relative humidity (RH) did not deteriorated within two to three days. After that period percentage of normal seedlings started to decline whereas percentage of abnormal seedlings started to rice up. The curves of abnormal seedlings changes attained maximum between 8th and 9th days of treatment when a slow decrease occurred. The amount of total phenolics in seed exudates was estimated by Prussian blue and Folin Denis methods. Results obtained with both methods were in mutual agreement although change in phenolic contents during accelerated ageing were clearer and absolute numeric values were higher in determination by Prussian blue assay. The high correlation was obtained among the content of total phenolics in exudates and percentage of abnormal seedlings in the course of accelerated ageing. The present studies indicate that measurement of total phenolics in seed exudates may be suitable for estimation of seed quality.

129 - EVALUATION OF THE VIABILITY AND VIGOUR OF TOMATO SEEDS BY TETRAZOLIUM TEST. SANTOS, MAGALI A.O., MARCOS-FILHO1, JULIO & NOVEMBRE2, ANA DIONISIA L.C.

USP/ESALQ/LPV, CP 09, 13418-900 - Piracicaba/SP, Brasil. [email protected] [email protected]

Procedures for the tetrazolium test to identify viability and vigour of tomato seeds (Lycopersicon lycopersicum L.) were studied in this research. First, seed viability was determined on four lots from cultivar Bruna. Seeds were imbibed between paper for two, three and four hours at 40°C or 45°C and then bisected longitudinally leaving seed intact at distal end. Staining was performed during one, two and three hours in a 0,075% tetrazolium chloride solution at 40°C. Tetrazolium seed viability was compared to results of germination and seedling emergence tests. The tetrazolium test to evaluate seed vigour was performed using cultivars Debora Max and Acclaim each represented by three lots. Interpretation considered three categories (high vigour, low vigour and non-viable seeds); the results were compared to those of germination, seedling emergence, accelerated aging and controlled deterioration tests. The best results were obtained with the combination of three hours imbibition at 45°C (seed water content from 44,8% to 46,4%) and staining for three hours at 40°C. The tetrazolium test was considered as a consistent option for the evaluation of viability and vigour of tomato seeds.

90 Budapest, Hungary, May 17th – 19th, 2004

130 - EFFECT OF DROUGHT CAUSED STRESS ON THE QUALITY AND YIELD OF SOYBEAN SEED TATIĆ, MLADEN, BALEŠEVIĆ-TUBIĆ, SVETLANA, MILADINOVIĆ, JEGOR & ĐORĐEVIĆ, VUK

Institute of Field and Vegetable Crops, Novi Sad, Maksima Gorkog 30, 21000 Novi Sad, Serbia and Montenegro

Stress caused by unfavorable environmental conditions during soybean seed (Glycine max (L.) Merr.) production can later influence the quality of the seed. Seed germination and seed vigor are significant traits of quality of the seed which depend on agroecological factors. Three varieties: Afrodita, Balkan, and Vojvodjanka belonging to different maturity groups were used in this study. The seed was produced at Rimski Sancevi (with no irrigation), and Srbobran (with irrigation). The study was conducted in three years differing in climatic conditions: favorable year, year with extreme drought and high temperatures, and year close to drought one. Obtained results showed that stress caused by drought, and high temperatures significantly decreased quality of soybean seed (energy of germination, germination, mass of 1000 seeds, and purity). Severe drought during soybean production exerted greater influence on decreasing the yield of soybean than the quality of seed. Irrigation increased the yield, and the quality of soybean seed, which pointed out the possibility of using irrigation for lowering the effect of stress caused by long-term drought.

131 - PHYSIOLOGICAL POTENTIAL EVALUATION IN MELON SEEDS (CUCUMIS MELO L.) TORRES, S.B.1 & MARCOS-FILHO2, J.

1EMPARN, P.O. Box 188, 59020-390, Natal, RN, Brazil ([email protected]) 2USP/ESALQ, P.O. Box 9, 13418-900, Piracicaba, SP, Brazil ([email protected])

This research was carried out to study procedures for controlled deterioration, accelerated aging, electrical conductivity, and potassium leakage tests, to establish their efficiency for identification of different levels of vigour in melon seed lots (Cucumis melo L.). The hybrids “AF-646” and “AF-682” each represented by five seed lots were submitted to tests of germination, seedling emergence, and seed moisture content determination. Results indicated that the controlled deterioration and the accelerated aging tests, both traditional and saturated salt accelerated aging, provided consistent identification according to their physiological potential, producing results comparable to those of germination and seedling emergence tests, thus demonstrating their sensitivity for vigour evaluation of melon seeds. On the other hand, the electrical conductivity test did not show to be an alternative for melon seed vigour evaluation, while the potassium leachate test needs further studies to adjust the methodology, including considerations on possible genotype effects.

132 - SEED VIABILITY, VIGOR AND STORAGE POTENTIAL OF GARDEN CRESS (LEPIDIUM SATIVUM L.). VERMA, S.S., KUMAR, R., VERMA, P.K. & DAHIYA, B.S.

Department of Seed Science & Technology, CCS Haryana Agricultural University, Hisar- 125 004; INDIA

The Garden cress (Lepidium sativum L.), also known as water cress or chandrasur is a cruciferous (Rabi), errect herbaceous glabrous annual plant of great medicinal value. The seeds, beside the propagating material, also useful in curing a number of deadly diseases like leprosy, scury, dysentry, diarrhoea, dyspepsia, leucorrhoea, asthma, cough, skin diseases, seminal weakness etc. Seeds can also be administered to cause abortion. Therefore, the crop is of immense importance to grow at commercial scale and hence the present study was undertaken to determine seed viability, vigor and storage potential. Results predicted high degree of aliveness by the dehydrogen-enzyme activity (viability test). The standard germination was tested in Between the paper (BP), Top of the paper (TP) and Quartz sand (S) and at three different temperatures (20°C, 25°C and 30°C). High

91 27th ISTA Congress – Seed Symposium germination per cent was recorded in BP (97.33%) followed by S (89.33%) at 20°C temperature for seven days. Minimum germination percent was recorded in TP (87.33%) method. The seedling vigor index was also found maximum in BP and S substrata, respectively. Results also showed high germination in TP (99.33%) method after giving the substrata treatment of 0.2% KNO3. It showed that the treatment of 0.2% KNO3 enhanced the vigourness of the seedlings. The stress tolerance capacity of the seed was determined by placing the seed in accelerated ageing chamber at high temperature (40±1°C) and high relative humidity (100%). Less germination was recorded in BP (30.00%) and TP (38.00%) substarta even after 24 h of accelerated ageing period. The germination potential of seed declined drastically and it confirms that the Garden cress seed cannot withstand the adverse conditions like other cruciferous crops, and hence, it is not recommended for long storage period.

133 - STUDIES ON VIABILITY AND VIGOR IN FENUGREEK SEEDS STORED UNDER AMBIENT CONDITIONS VERMA, SHER SINGH, KUMAR, SANDEEP & KUMAR, RAMESH

Department of Seed Science & Technology, Chaudhary Charan Singh Haryana Agricultural University , Hisar-125004, Haryana-INDIA, [email protected]

42 seed lots of fenugreek (Trigonella foenum-graecum L.) comprised of three seed lots of 14 genotypes were analyzed for standard germination (%), seedling length (cm), seedling dry weight (mg), seedling vigor index-I, accelerated ageing test, tetrazolium test (%), dehydrogenase activity test, pH exudates test, electrical conductivity test (µ mhos/cm/seed) and emergence rate index. Results revealed that the germination percent of all the 14 genotypes remained above the Minimum Seed Certification Standard (70%) limit in six months and 18 months old seeds. Significant decline in germination, seedling length, seedling dry weight, seedling vigor index-I, tetrazolium test, dehydrogenase activity test, accelerated ageing test, pH of seed leachates and emergence rate index was observed as the age of seeds increased, however this decline was more rapid in 2½ year old seeds stored under ambient conditions. Electrical conductivity increased significantly with increase in the age of the seeds. Germination percent, seedling length, seedling vigor index-I, seedling dry weight, dehydrogenase activity test, tetrazolium test, accelerated ageing test, pH of seed leachates were positively and significantly correlated with emergence rate index. Results also revealed that the germination percent, seedling dry weight and dehydrogenase activity tests were found reliable predictors of emergence rate index. So, these parameters can be used as reliable predictors of field performance.

134 - SEED MATURITY AND HETEROCARPY ON SEED QUALITY IN ZINNIA VIJAYAN, R., NATESAN, P. & MURUGESAN, P.

Tamil Nadu Agricultural University, Seed Science and Technology Laboratory, Laley Road, Coimbatore- 641003, India, [email protected]

Zinnia elegans L. is mostly grown in the controlled condition in major part of the world. In India, it is possible to grow commercially in the field condition. But seeds show slow and protracted germination. Hence, stages of full maturity, heterocarpy of achene (size, shape and colour) were investigated to know the correct harvest time and germination improvement, respectively. Results indicated that the physiological maturity occurred 40 days after anthesis. Seeds retained in British Sieve Size (BSS) 6 x 6 was found to give superior seed quality characters. BSS 8 x 8 was found optimum with maximum seed recovery and satisfactory seed quality. Achenes of Z. elegans can be separated into four categories as oblong light brown, oblong dark brown, linear dark brown and linear light brown based on pericarp colour and shape. Significant seed quality differences are observed among above categories.

92 Budapest, Hungary, May 17th – 19th, 2004

135 - COMPARISON OF THREE METHODS OF MOISTURE CONTENT ADJUSTMENT: THEIR IMPACT ON GERMINATION AND VIGOUR TESTING WAGNER, M. HELENE, PREVEAUX, ANNE, BEAULATON, MATTHIEU & DUCOURNAU, SYLVIE.

GEVES-SNES, rue Georges Morel, B.P. 24, 49 071 Beaucouzé Cedex, France [email protected]

The three methods for raising a required moisture content (MC) recommanded by the ISTA Vigour Handbook were compared at 3 MC levels and on 4 species : bean, pea, soyabean and rapeseed. These methods differ in the speed of hydration for seeds : the quickest one (few minuts) is the water added (WA) method using a micropipette to add water missing in the seeds to reach the required MC, followed by the filter paper (FP) method (few hours) where seeds are imbibed on a moist paper till they reach the required MC. The last one is the high relative humidity (HRH) method which is the most equilibrate method for seeds but consuming time (few days or weeks). It consists in placing seeds in an incubator with high relative humidity and low temperature till they reach the required MC. The 3 methods were firstly compared in their accuracy to give the required MC: the FP method gave the best results for any moisture content level. When required MC increased, the accuracy of the three methods decreased especially for HRH method. Then, methods of adjustment were compared upon the results they provided on different quality tests: germination, conductivity and controlled deterioration (accelerated ageing for soyabean) tests. Germination test was independant from the MC adjustment method for any specie, except for some bean seed lots. The three methods were equivalent on pea seeds for any quality testing. For the other species, WA was the most penalizing method and affected vigour results.

136 - CONDUCTIVITY TEST ON DIFFERENT SPECIES AMONG THE MOST IMPORTANT CROPS IN FRENCH AGRICULTURE. WAGNER, M. HELENE, PREVEAUX, ANNE, MOIZAN, ELISE & DUCOURNAU, SYLVIE

GEVES-SNES, rue Georges Morel, B.P. 24, 49 071 Beaucouzé Cedex, France [email protected]

Bulk conductivity test on garden pea was introduced in ISTA Rules in January 2002. To get a larger useful test, we try to extend it to other species among the most cultivated in the French production: wheat, maize, rapeseed, sunflower, and lucerne. Five species were chosen because of their different storage tissues: oil crops, starchy ones and legume and because they represent different kind of botanical seeds: akene, caryopsis and single seed. Five samples were used by specie and analysed with a conductivity meter fitted with a dip cell having a cell constant 1 cm-1. The temperature of the conductivity test was fixed at 20°C like for pea but the other parameters of the test were adjusted to the size of samples and to the kind of seed storage tissues. Finally, three methodologies can be suggested for rapeseed, wheat and maize, which are accurate and repeatable and which can classify seed lots more tightly than germination test. They should now be validated with comparative tests between different laboratories and with field emergence results. Some work is still to be done on sunflower because of the electrolytes leakage from the pericarp more or less important belong to the variety. For lucerne seeds, so much variability is met with hardseededness and to solve this problem, recovery and weighing of hard seeds is not enough.

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137 - AN ACCURATE GERMINATION TEST FOR FIELD EMERGENCE PREDICTION OF SOYBEAN SEED WONGVARODOM, V. SANTIPRACHA, W. & SANTIPRACHA, Q.

Department of Plant Science, Faculty of Natural Resources, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand, [email protected]

An accurate germination test was studied to evaluate the field emergence of soybean seed. Seven varieties of soybean namely CM. 2, CM. 60, NS. 1, RM. 1, SJ. 4, SJ. 5, and ST. 2 seed of various qualities were used. They were subjected to tests of standard germination and field emergence. An accurate germination test was done under room conditions by planting the seed in 1,000 g of soil in plastic baskets size 19x26x6 cm at 1 cm depth and watering at 70% of plant available water every 2 days. The germination evaluations were done at 3 days. Three soil types: sandy loam, silt loam, and clay were used in the test. Germination percentage, speed of germination, seedling growth rate as shoot length and shoot dry weight were measured. The accurate germination test had a high correlation (r=0.914-0.976) with field emergence of all varieties of all soybean seed qualities in the three soil types.

138 - FDA ANALYSIS OF KENYAN AND UK ORCHID SPECIES WOOD1, CHRISTOPHER B., MUGAMBI2, GEORGE K. & PRITCHARD1, HUGH W.

1Seed Conservation Department, Royal Botanic Gardens Kew, Wakehurst Place, Ardingly, West Sussex, RH17 6TN, UK. Email: [email protected] 2National Museums of Kenya, East African Herbarium, P.O. Box 40658, Nairobi, Kenya

The vital stain fluorescein diacetate (FDA) was used to assess the viability of a range of orchid species. For 10 UK orchid species, in vitro asymbiotic germination was compared with FDA viability assessments. In total, 23 seed lots were tested, and demonstrated a linear relationship between germination and FDA viability, with a slope close to unity. In addition, the pattern of seed viability loss during artificial ageing at 60°C was monitored in one species, Dactylorhiza fuchsii, and found to be similar, though not statistically identical, when measured by germination and FDA staining. The test was also used to assess the viability of an additional 26 UK species and nine Kenyan species. In contrast to the UK species, FDA substantially under-estimated the viability of Kenyan species tested, due to problems with stain permeability and embryo pigmentation. Thus, whilst the FDA test can provide a dependable and accurate measurement of viability in UK orchids, further work is needed before it can be adopted for seed viability testing throughout the Orchidaceae.

139 - EVALUATION OF COTTON SEEDLING VIGOUR IN OSMOTIC STRESS ZANGI, M.R.

Board of scientific, Cotton Research Institute of IRAN, [email protected]

Environmental stress decreased crop production about 71 percent. Drought stress is the most limitation factor for agriculture production in the arid and semiarid regions. For this reason, investigation on various aspects of drought on species and drought resistance improvement are much important. This study was conducted to evaluate 12 cotton genotypes resistance to osmotic stress. Osmotic treatments were 0, -2, -4, -6, -8 bar. The experiment design was conducted as a factorial experiments in completely randomized design . The first level was osmotic stress and the second levels were 12 genotypes. Variables were seedling dry mater, seedling fresh weight, normal seedling number, radicle and hypocotyl length and germination rate. An increase in osmotic stress up to –8 bar caused a increasing in the germination rate. -8 bar of osmotic stress had the highest germination rate and normal seedling number. Genotypes

94 Budapest, Hungary, May 17th – 19th, 2004 had different level of seedling vigour in osmotic stress. Sahel genotype had the highest germination rate.

140 - STUDY OF IRANIAN LANDRACE OF COTTON GENOTYPES RESISTANCE TO SALINITY IN GERMINATION STAGE ZANGI, M.R.

Board of scientific, Cotton Research Institute of IRAN, [email protected]

About 40 percentage of 220000 ha of cotton farms challenge germination in Iran. Yield decreased 20-30 percent in salinity soil. Improvement of saline soil has a lot of cost and time. The most approach is crop breeding for salt resistance. This study was conducted to evaluate 12 cotton genotypes resistance to salinity in the germination stage. Salinity treatments were 0, 5, 10, 15 and 20 ds/m). The experiment design was conducted as a factorial experiments in completely randomized design. The first level was salinity stress and the second levels were 12 genotypes. Variables were seedling dry mater, seedling fresh weight, normal seedling number, radicle and hypocotyl length and germination rate. An increase in salinity up to 20 ds/m caused a reduction in the seedling fresh weight, radicle length, hypocotyl length and germination rate, but to increase seedling dry matter. Genotypes had different level of resistance to salinity stress. Garmsar landrace genotype had the highest seedling dry matter and radicle length and hypocotyl length. Varzaneh Jozagh landrace genotype had the highest germination rate.

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4. Seed Systems in Emerging and Developing Economics Topics: Development and status of seed systems; seed quality management; evaluation of seed quality; role of informal and formal seed sector

141 - PROBIT MODELLING OF SEED PHYSIOLOGICAL DETERIORATION IN HUMID TROPICAL SEED STORES DANIEL, I. O.

Department of Plant Breeding & Seed Technology, University of Agriculture, PMB 2240, Abeokuta, Nigeria, [email protected], [email protected]

In a preliminary trial to model seed deterioration in tropical seed warehouses, two survival data sets of maize and soybean seeds stored under the prevailing ambient room conditions of the humid tropics were subjected to PROBIT analysis. The analysis of individual seed survival data for all seed lots showed non-significant χ2 (p>0.05), suggesting that seed deterioration under the humid tropical seed stores will follow the normal distribution pattern as earlier described by the Ellis and Roberts (1980) seed viability model. Estimates of seed longevity parameters like the slope of the seed survival model were negative showing that seeds were systematically deteriorating over the three months storage period. The tolerance distribution of the year-2001 data estimated from the probit procedure gave µ of 114.7 days and σ of 92.1 days for maize seeds, µ of 43.8 days and σ of 60.8 days for soybean seeds respectively, the µ representing time to 50% seed viability loss while σ representing time to lose 1% seed viability under storage. The results showed significant differences in seed deterioration rates and ultimate longevity between the two different species. Initial seed quality before storage also resulted in differences in rates of seed deterioration, high initial seed quality positively influenced seed deterioration and ultimate longevity in storage. Model parameters for predicting seed longevity under tropical seed warehouse conditions were estimated for seed lots with initial germination within those of the seed lots used in the trial.

142 - SEED PRODUCTION AND ANDRO-STERILITY IN NORMAL AND QUALITY PROTEIN MAIZE (QPM) IN MEXICO ESPINOSA-CALDERÓN1, ALEJANDRO, TADEO-ROBLEDO2, MARGARITA, GÓMEZ-MONTIEL3, NOEL, SIERRA-MACÍAS3, MAURO, REYES M.3, CESAR, CABALLERO H.3, F., PALAFOX C.3, A., BETANZOS M3., E. & COUTIÑO E3., B.

1Seed Production Program, National Research Institute of Forestry, Agriculture and Livestock (INIFAP, Mexico). [email protected] 2National Autonomous University of Mexico (UNAM, Mexico). [email protected] 3Maize Program, National Research Institute of Forestry, Agriculture and Livestock (INIFAP), Mexico.

Mexico has the potential to produce some 270,000 hectares with quality protein maize. Since 1998, a significant number of researchers from the National Research Institute of Forestry, Agriculture and Livestock (INIFAP), have been working with CIMMYT researchers to obtain some QPM varieties and hybrids. As a result of this effort, some high-yielding materials with good agronomic characteristics have been developed. But it has proven to be much more difficult to convince farmers to use the seed of the new materials. In 1999, INIFAP released 26 hybrids and varieties, but only a few of these are used. One important problem is that the National Seed Producing Company (PRONASE) is bankrupt. One possibility to increase the seed adoption rate of QPM hybrids is to use andro-sterility. INIFAP researchers have been working for many years with the progenitors of the hybrids of normal quality. Three new hybrids with the andro-sterile scheme have been released recently, and mixes (H-48 AE, H- 50 AE, H-153 AE). The new andro-sterile version of those hybrids has better yield (15%) again the initial version. It is necessary to have the complete andro-sterile scheme. Work is currently under way to achieve the complete andro-sterile scheme, as well as to identify and develop inbred lines that have the fertility restoring capacity. In the case of QPM hybrids,

96 Budapest, Hungary, May 17th – 19th, 2004 work started in 1999, by introducing andro-sterility to the basic lines, and by trying to identify restoring lines. Progress has been achieved, and now the program has second and third inbreds towards achieving quality protein in elite lines. Restoring lines have also been identified, and these are used as restoring donors to the QPM lines.

143 - A CONTRIBUTION TO THE DEFINITION OF BATCH SIZE – CONTROL UNITS AND MINIMUM AVERAGE SAMPLE FOR ANALYSIS OF THE SEEDS OF SOME FOREST TREE SPECIES IN CROATIA GRADEČKI, M. & POŠTENJAK, K.

In Croatia forest seed husbandry has a long tradition which is interwoven in written documents and legislation and conditions inherited – raised pine cultures severalcenturies old. The area of Croatia is extremely diverse, and although it only covers 0,59 % of the area of Europe, 4 500 ( 36,93 %) of the plant species in Europe (totalling 12184) can be found here. However, today we are witnessing the drastic consequences of disruption of forest ecosystems, which can be seen in forest seed husbandry, by the increasing need for greater amounts of seeds which are necessary for supplementing natural reforestation. In order to satisfy as far as possible the demand for seeds and at the same time to protect the disrupted forest ecosystemis rational managment of seeds is necessary, during which it is particularly important to preserve the biodiversity of paricular species. The law in Croatia differentiates selected seed and normal seed from acknowledged seed stands. Acknowledged seed stands are phenotypically the best parts of the forest for certain species which are separted within a certain seed unit in order to satisfy the need for seed of certain species in the area of a particular seed unit. In Croatia there are three districts with 79 ecological – management types, which are a good guideline for definition on of seed units and consequently their area (the average area ranges from 250 to 69,250 ha). The size of the area of acknowledged seed stands in particular seed units depends on several parameters, among which the following are important: mixsture ratio (according to the number of trees) of seed species, relation of normal and acual number of trees by ha, average age of stands, average degree of crown defoliation, periodic full seed crop, and success of natural reforestation (average optimal area in a certain seed unit ranges from 0,003 % to 20.0 % of the area of the seed unit, i. e. 5 to 210 ha, and the average annual production of forest seed in a certain seed unit ranges from 13,75 kg for Spruce to 10 086 kg for Peduncled Oak). The batch of forest seed should be gathered from the area of one seed unit, i. e. there may be more, but on the whole it should be a guarantee of protection of the biodiversity and preservation of the particular ecotype of individual tree species. Croatian legislation and standars in forest seed husbandry, with regard to batches and minimal samples, differ significantly from ISTA standards. This difference is conditioned by several specific parameters which have determined these differences (differences are from 0 to 2 000 %) and which function as protection of the biodiversity of a certain species and rational use of seed.

144 - AN ALTERNATIVE APPROACH TO GERMINATION TESTING BASED ON BAYES THEOREM JACKISCH, W.

Sächsische Landesanstalt für Landwirtschaft, Waldheimer Str. 219, DE 01683 Nossen, Germany, [email protected]

Bayesian methods have gained increasing popularity in different fields as an alternative to classical statistical methods in the last few decades. In this paper the hypothesis is tested whether Bayesian estimation methods could be used to reduce the sample size in the traditional 400-seed germination test. The basic idea for this approach is the incorporation of Monte Carlo estimates into a reduced seed germination test. In a simulated germination study with 10.000 runs an identical frequency distribution of the germination results from each replication was observed. By using Bayes theorem the probability distribution for larger

97 27th ISTA Congress – Seed Symposium sample sizes could be calculated from one or two replication distributions. In the case of germination assessment, when prior probabilities are unknown, the prior could be estimated from an initial sample. In the studies an initial sample size of 2 x 100 tested seeds has been chosen. The germination results of the initial samples were augmented by two binomial generated Monte Carlo estimates as substitute for the missing values of the traditional sample size. All results of the alternative procedure indicated a smaller distribution than the expected germination distribution of the standard method. Data sets from commercial seed lots have been used to check the findings. This proposed alternative procedure for germination assessment confirmed identical results in both the simulation study and the collected data sets of commercial seed lots. All observed distributions are computable by the Bayes theorem. The posterior distributions from the reduced 200-seed germination test augmented by two generated Monte Carlo estimates showed deviations similar to 800-seed tests caused by the conditional probability of the initial sampling. Thus, the alternative approach seems to have a great potential for the required point estimation in germination testing.

145 - EESNET – AN EASTERN EUROPEAN BRIDGE TO THE GLOBAL SEED SECTOR KŘEHLÍK, LUDĚK

CMSSA - Czech Seed Trade Association, Antala Staska 30, 140 02 Praha 4, Czech Republic phone +420 222 135 295, fax +420 222 135 294, [email protected]

Disintegration in the region of central and Eastern Europe has been a notable feature not avoiding the seed sector. Regardless the direct economical needs of the countries, the Czech Seed Trade Association (CMSSA) developed an effort to assist the countries of the region in collection of the information from the global seed sector development and development of contacts with the rest of the world. That effort resulted in establishment of the Eastern European Seed Network (EESNET) in 2000. EESNET has been recognised by all international seed organisations as a useful tool to share the information in the region and they participate via their supreme representatives each year in the Annual Meetings organised by EESNET. Those meetings are attended well by people from private sector as well as governmental institutions. EESNET is now a group of 9 countries and after 4 of them will join EU in 2004, it will probably continue its activity in a more extended framework, depending on a decision of the countries those will be active within the group. The priority of the future EESNET mission should be to extend the collaboration with the international seed organisations, especially in European level, in order to establish the trustful environment in the region.

146 - PROPOSAL FOR THE REHABILITATION OF THE SOUTH SEED LABORATORY BY WESTERN OR EUROPEAN (NORTH) SEED LABORATORY “IN CASE SENASEM SEED LABORATORY IN BUKAVU SOUTH – KIVU PROVINCE IN DEMOCRATIC REPUBLIC OF CONGO”. MAZAMBI BYAKOMBE, JONATHAN

SENASEM’s Provincial Coordinator and their Technicians

(no summary)

98 Budapest, Hungary, May 17th – 19th, 2004

147 - SEED TESTING OF UNSHELLED SEED LOTS OF GROUNDNUT (ARACHIS HYPOGEA L.) MITI, FRANCISCO

SCCI, Box 350199, Chilanga, ZAMBIA. Fax. 260-1-278170, E-mail: [email protected]

Groundnut (Arachis hypogea L) is one of the major food and cash crops for small-scale farmers in Zambia. However, increased production of the crop is hindered by lack of good seeds. Certified seed are less common for the crop due to high seed loses. The international rules for seed testing of the International Seed Testing Association (ISTA) provides that a shelled groundnut seed lot is tested and therefore marketing is done on shelled seed lot. Shelled seed lots experience a lot of seed damage and lose viability quickly thereby discouraging seed business. Such difficulties are delaying the growth of the groundnut industry as introduction of better seeds from other areas is frustrated by huge loses incurred in the transportation. In the tropics, high temperatures too accelerate the deterioration of the groundnut seed viability particularly when shelled. This paper proposes an alternative procedure for the testing of unshelled groundnut seed lot. The unshelled groundnut seed lot is sampled and purity analysis done on the unshelled sample. The pure seed fraction of the sample is then shelled and the shelling percentage determined. The shelled nuts become a working sample for the germination test following the current ISTA rules. The tested unshelled seed lot can thus be certified for sale under respective national regulations.

148 - THE DEVELOPMENT OF THE COTTAGE SEED INDUSTRY IN ZAMBIA MITI, FRANCISCO

SCCI, Box 350199, Chilanga, ZAMBIA. Fax. 260-1-278170, E-mail: [email protected]

The majority of Zambia’s farming community is small-scale and is locate in rural areas where seed availability is poor due to poor infrastructure. They also lack cash to buy seed and depend on local and unimproved seed which yields poorly. To promote agricultural production the government of Zambia and its cooperating partners initiated the development of the cottage (rural) seed industry in about mid 1990s for a sustainable seed delivery system in rural areas. Seed quality control has been decentralized and the private sector participates in it by way of licensing while the Seed Control and Certification Institute (SCCI) monitors the licensing scheme. Licensees inspect and sample seed lots that are tested mainly by satellite laboratories that have since been established. Foundation seeds are obtained from public research while the government extension service guides farmers in seed production. In all provinces, smallholder seed growers multiply and market improved seeds within their communities. During the 2000/2001 season, seed enough for about 10% smallholder farmers was produced by smallholder seed growers. The seeds are packaged in factory labelled plastic bags and are mainly sold to the local faming community. Some seeds change hands by the barter system. Seed provision has become a major source of income for seed growers. Other regular activities include seed promotions, training, awareness on gender and HIV/AIDS.

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149 - EVALUATION OF MECHANICAL DAMAGES IN SOYBEAN SEEDS BY IMAGE ANALYSIS1 OBANDO FLOR2, EBERT PEPE, CICERO3, SILVIO MOURE, DE BARROS FRANÇA NETO4, JOSÉ & KRZYZANOWSKI4, FRANCISCO CARLOS

1Part of the thesis presented for the first author for attainmment of the heading of Doctor in Agronomy 2Eng. Agr., PhD; Departament of Seeds – National Institute of Agricultural Research (INIA-Lima - Perú) [email protected] 3Eng. Agr., PhD; Departament of Crop Science, ESALQ/USP, CP 09, 13418-900 Piracicaba-SP-Brazil 4Eng. Agr., PhD., EMBRAPA- (CNPSo). CP 1061, CEP 86001 Londrina, PR -Brazil

The present work had the objective of developing a non-conventional method for determination of mechanical damage in soybean seeds, and also to verify its relation to the physiological quality, using the technique of image analysis. Unlike to the traditional tests, this technique provides the analysis of seeds without destroying them, and together with germination test, it can be observed that the damages detected in the image analysis is really important because of their influence in the germinative process. The research was carried out with three cultivars of soybean (FT-2, FT-10 and IAC-2), possessing different contents of lignin in the seed coat (different behaviors to the resistance for mechanical damages). The mechanical damages had been caused by means of the pendulum test in six different regions of the seeds. The image analysis identified external and internal mechanical damages was made by means of image analysis and by means of tetazolium test.The image analysis identified external and internal mechanical damages in soybean seeds, and also, was efficient to detect weathering and stink-bug damages. The image analysis presented a good correlation will the tetrazolium test when it was considered the evaluation of the mechanical damage and the viability of the seeds.

150 - EVOLUTION OF SEED QUALITY IN WHEAT AND CORN SOWN IN PAMPA REGION OF ARGENTINA, DURING THE AGRICULTURAL PERIOD 1993-2002 . *PERETTI, A., CINALLI,C., BERRUET, H. & BARIFFI, H.

Unidad Integrada Facultad de Ciencias Agrarias-Estación Experimental Agropecuaria INTA Balcarce C.C. 276. (7620) Balcarce. Argentina, [email protected], tel-fax : 0054 2266 439141

Argentina is the fifth world producer of wheat and corn, the fourth world exporter of wheat and the second of corn. The surface sown with wheat is 6.000.000 ha and 3.000.000 ha with maize. The southeast of Buenos Aires province, in the pampas region, contributes with 30% to wheat national production (14.000.000 tons), and 5% to maize (15.000.000 tons) The Seed Analysis Laboratory of Unidad Integrada Balcarce receives samples from an area of 4 million hectares, and seed quality diagnosis reflects the trend of seed market. In this study the evolution of wheat and maize seed quality sown in this area during the period 1993 – 2002 was estimated. The samples were processed by standard germination, accelerated ageing, cold test (ISTA), and by methanol at 20%. The samples of wheat evaluated, mainly Triticum aestivum L.,were 2355. Mean standard germination of each crop season ranged from 86% in 1995 to 93% in 1999 and 2001 .The maximum of samples below the official level of tolerance (germination 85%) was registered in 2002 (26%), the minimum (4%) in 1999. Only 20% of sown wheat were certified seeds.Samples of Zea mays L. were 1684. The highest annual mean germination reached 97% in 1998 and 2001, the lowest 88% in 1993.The percentage of samples below the tolerance (90%) ranged from 35% to 3%. Vigour tests showed the increasing quality of seed, with high vigour in more than half of the samples analyzed. Certified seeds were sown in almost 100% of sowing surface.

100 Budapest, Hungary, May 17th – 19th, 2004

151 - Maize seed production at the Agricultural Engineering Department of the National Autonomous University of Mexico (UNAM) TADEO-ROBLEDO1, MARGARITA, ESPINOSA-CALDERÓN2, ALEJANDRO, MARTÍNEZ- MENDOZA1, RAFAEL & PIÑA DEL VALLE3, ANGEL

1National Autonomous University of Mexico (UNAM), Mexico, [email protected] 2Seed production Program, National Institute of Forestry, Agriculture and Livestock Research (INIFAP, Mexico) [email protected], 3Ceres de Mexico, Seed Production Company.

Since 1984, the Agricultural Engineering Department of the Cuautitlan Superior Studies Institute – National Autonomous University of Mexico (UNAM), with the support of CIMMYT and other institutions, has developed a strong maize plant breeding, seed production and seed technology program, oriented towards the development of highland (2200-2600 masl) and transition (1800-2200 masl) environments. For these environments, nine materials were released in the period 1995-2003, with high yielding and other good agronomic characteristics. Initially, the materials were reproduced by the National Seed Production Company (PRONASE); however, this scheme failed as PRONASE underwent several restructuring phases and is currently bankrupt and in the process disappearing. Under these circumstances, a new scheme for seed production is needed, and one alternative is small- scale, private seed companies and seed producers’ associations. With the assistance of students from the Seed Production and Technology class, and from those doing social service, field study, or thesis prior to graduation, new inbred lines and the best combinations of three-way hybrids and OPVs are being evaluated. The main objective is to obtain good maize materials as favorable alternatives for highland and transition zones in México, and to help increase the use of improved seed in these environments, currently at very low levels of 6% y 37% respectively. In both regions, UNAM materials compete favorably with materials from private seed companies, and we expect that when more and better materials are available, improved seed can be put in farmers’ hands at fair prices and quality seed.

152 - CHARACTERIZATION OF RAPESEED, MUSTARD AND TARAMIRA CULTIVARS USING MATURE PLANT CHARACTERS YADAV1, SHIV KUMAR, YADAV2, T.P. & RAM3, C.

1Division of seed science and Technology, IARI, New Delhi-110012, India 2Oil Seed Section, Department of Plant Breeding, CCS Haryana Agricultural University, Hisar – 125004, India 3Department of Seed Science and Technology, CCS Haryana Agricultural University, Hisar – 125004, India

Eighteen mustard (Raya) i.e. Brassica juncea (L), four rapeseed (toria) i.e. Brassica campestris (L). and one cultivar each of Brassica campestris var. Yellow Sarson, Brassica campestris var. Brown Sarson, Brassica napus (L). and Eruca sativa (L). (Taramira) were studied for 14 various quantitative and qualitative characters for two years. Mature plant characters like growth habit, mature leaf shape, mature leaf color, intensity of mature leaf color, stem pubescence, mature plant height, number of primary branches, number of secondary branches, main shoot length, morphotype uniformity and basal side shoot development were used to classify the varieties, which provided a sound classification system as these characters exhibit distinct varietal patterns. The mature leaf color, morphotype uniformity and plant height were the plant characters that showed continuous differences during both the seasons. Hence, all the other characters studied were found useful for the purpose of cultivar identification. A list of key characters useful in variety identification was also prepared. It was possible to distinguish all the cultivars individually using the seed key.

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5. Seed Lot Hygiene Topics: weed contamination; seed-borne pathogens, pests; endophytes, phytosanitary standards

153 - BIO-DETERIORATION OF FENNEL (FOENICULUM VULGARE MILL.) SEEDS DUE TO SEED BORNE FUNGI AGRAWAL, MUKTA & SAXENA, SHUBHRA

Department of Home Science, University of Rajasthan, Jaipur 302 004, India

Fennel (Foeniculum vulgare Mill.) is an important seed spice grown in India. Rajasthan is one of the leading States in its production. It is commonly used as mouth freshener, flavourant and has rich medicinal properties. To study the biodeterioration of fennel seeds due to micro- organisms, 122 seed samples were collected from the various Agro-climatic zones of Rajasthan State. Dry seed examination revealed besides the asymptomatic seeds, the presence of moderately and heavily discoloured seeds affecting the seed quality adversely. A total of 92 fungal species of 39 genera were recorded in seed samples with varied incidence in moistened blotter test. Species of Alternaria, Aspergillus, Chaetomium, Curvularia, Fusarium, Phoma and Trichothecium were dominant. Biochemical studies of seed samples of fennel naturally infected with species of Alternaria, Aspergillus, Cercospora, Curvularia, Drechslera, Fusarium, Phoma and Trichothecium individually or in various combinations caused a significant change in total carbohydrate, protein, lipids, fibre, phenol and volatile oil contents of seeds as compared to the healthy seeds.

154 - EFFECT OF SEED- BORNE INFECTION OF FUSARIUM OXYSPORUM SCHLECHT. EX FR. ON SEED QUALITY OF FENNEL SEED AND ITS MANAGEMENT AGRAWAL, KAILASH & DWIVEDI, MANISHA

Botany Research Lab., P.G. Department of Botany, Agrawal P.G. College, Jaipur 302 003, IN- INDIA [email protected]

A study of 127 seed samples of fennel belonging to 21 districts of Rajasthan State, India revealed F. oxysporum in 58 samples from 16 districts with 1-51% incidence. Histopathology of naturally infected seeds revealed the pathogen localized to carpophore, pericarp and seed coat only in asymptomatic seeds whereas in carpophore, pericarp, seed coat, endosperm in moderately and heavily discoloured seeds and in embryo also in heavily discoloured seeds. The mycelium was inter-as well as intracellular .The pathogen caused pre-and post emergence losses and produced seedling symptoms. In general , a significant decrease (P<0.05 and P<0.01) in moisture , crude fat , total soluble sugars , crude protein and volatile oil and increase in crude fibre and total phenols was recorded in samples naturally infected with F. oxysporum. Microwave treatment at 40 and 60 power levels for 5 min were found significantly effective in controlling the pathogen and also improved seed germination. Among the three fungal antagonists viz. Trichoderma harzianum, T. viride and Gliocladium virens used to control the pathogen, Trichoderma harzianum caused maximum inhibition but it also caused severe loss in seed germination, hence, G. virens was recommended to be the most suitable one.

102 Budapest, Hungary, May 17th – 19th, 2004

155 - COMBINED EFFECTS OF SEED TREATMENT WITH CARBOXIN-THIRAM AND INSECTICIDE ON GERMINATION AND SEEDLING DAMPING -OFF DISEASE IN COTTON AZADDISFANI, F. & NARAGHI, L.

Board of scientific, Cotton research Institute of IRAN, [email protected]

The cotton is one of the most important crop plants. Different control methods are recommended for Pathogens and pests (especially Trips) in seedling stage. Commonly the most control method is seed Treatment with fungicide and insecticide combined application. On regarding to reports about unfavorable effects on seed germination, it is needed that these effects are studied on Sahel seed Treatment with Carboxin –Thiram (common fungicide in Iran) and insecticides (Larvin and Gaucho) in laboratory and greenhouse with infected soil (field soil) and sterile soil. These experiments were conducted indivisual and factorial. The first factor was dose levels (maximum, medium, minimum and without Carboxin-Thiram,) and second factor was insecticides (Without insecticide, Gaucho and Larvin). Experiments were conducted in complete randomized design in laboratory and greenhouse. Studied characters were included of germination rate I laboratory, main root length, number of lateral root, height, dry weight, root dry weight (after 40 days), seedling damping-off, emergences rate (after 12 days) in greenhouse. Laboratory study results indicated that germination rate was non significant. In sterile soil, all of traits were significant (except seedling damping -off). Application of medium dose wit gaucho increased total number of lateral root. On the other hand, larvin without fungicide reduced this character. In infected soil , main root length, seedling damping-off and emergences rate were significant. Application of minimum dose (Carboxin-Thiram) without insecticide controlled seedling damping-off disease. This treatment had the most of emergences rate. So application of minimum dose was caused inciting and increase of emergences rate.

156 - GREENHOUSE AND FIELD EVALUATION OF SEED TREATMENT CHEMICAL IN RELATION TO ROOT CHARACTERS IN COTTON AZADDISFANI, F. & NARAGHI, L.

Board of scientific, Cotton research Institute of IRAN, [email protected]

In this research, influence Sahel seed treatment with Baytan and insecticides is studied on root parameters that included of main root length, lateral root number and lateral root length in first, second, third and forth section of main root, root dry weight in field (Gorgan) and greenhouse. Greenhouse experiments were included of infected soil (field soil) and sterile soil. These experiment were conducted individual and factorial. The first factor was dose levels of (maximum, medium, minimum and without Carbendazim) and second factor was insecticides (Larvin, Gaucho and without insecticide). Experiment was conducted in complete randomized design in greenhouse but experiment was conducted in randomized complete block design in field. Field results were showed that lateral root number, lateral root length in first, second and forth root section were significant. Application maximum dose of Baytan without insecticides caused increase of lateral root number (14.45) and medium dose combined with Gaucho was caused reduce of lateral root number. In greenhouse with infected soil, characters such as main root length , lateral root length in first and forth root section were significant. Application Gaucho without Baytan had the most of main root length (6.25 cm) and application medium dose of Baytan without insecticide had the least of main root length. In sterile soil, seed treatment with Baytan and insecticides was caused difference except lateral root length in second root section. Application medium dose of Baytan with Larvin increased lateral root number. Gaucho treatment without Baytan had the most of root dry weight and Larvin treatment with minimum had the least of root dry weight.

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157 - INFLUENCE OF CARBENDAZIM AND INSECTICIDE ON ROOT PARAMETERS OF COTTON SEEDLING AZADDISFANI, F.

Board of scientific, Cotton research Institute of IRAN, [email protected]

Stand establishment of cotton seedlings is one of the most critical stages in cotton production. Regarding to reports about unfavorable effects of seed treatment with fungicides and insecticides combined application on seed germination, in this survey effect of Sahel seed treatment with Carbendazim and insecticides were studied on main root length, lateral root number and lateral root length in first , second, third and forth section of main root, root dry weight in field (Gorgan) and Greenhouse. Greenhouse experiments were included of infected soil (field soil) and sterile soil. These experiment were conducted individual and factorial. The first factor was dose levels of (maximum, medium, minimum and without Carbendazim) and second factor was insecticides (Larvin, Gaucho and without insecticide). Experiment were conducted in complete randomized design in greenhouse but experiments were conducted in randomized complete block design in field. Results were indicated in field that combined application different doses of Carbendazim and insecticide were caused changes on lateral root number, lateral root length in first and second section. Application of maximum dose with Gaucho had the most of lateral root number and maximum dose of Carbendazim treatment with Larvin had the least of lateral root number. In infected soil (greenhouse), all of the characters were significant except lateral root number, lateral root length in root third section and root weight. However in sterile soil, every characters were significant. Control treatment had the most of lateral root number and the least of weight root, actually this treatment had weak roots, on the other hands maximum dose of Carbendazim (without insecticide) had the least of weight root. So all of the treatments were caused decreasing of lateral root number except Gaucho treatment.

158 - RELATIONSHIP BETWEEN SEEDLING VIGOUR AND DAMPING-OFF DISEASE DIFFERENT SPECIES OF COTTON AZADDISFANI, F. & ZANGI, M.R.

Board of scientific, Cotton research Institute of IRAN, [email protected]

Seedling diseases cause a decreasing yield and are usually the major disease problems in cotton production in the world. Several soil-borne fungicide are responsible; however, cultural and environmental factors that delay seed germination seedling growth make the problem more severe. Since that causal agent of the disease is a soil borne fungus, therefore, chemical control is not always effective. In addition to this, wide spread use of chemicals results in pollution of the environment and appearance of resistant races of the pathogen, need for the search of alternative methods to control this disease is clearly obvious. Genotypes of Damping-off resistance could be promising alternative method. In this study the possibility of use of seed-seedling vigour was investigated for selection damping-off resistance. genotypes were 4 different species cotton. Genotypes were Barbadence 5539, Siokra324, Sahel, Arboreum, Oltan, Giza, T-14, red leaf okra, Arya and Gokrova. Studied variables were germination rate, normal seedling number, radicle and hypocotyl length, radicle and hypocotyl fresh weight, radicle and hypocotyl dry weight, and damping- off of 10, 20, 30 days. Damping off was calculated to minus laboratory germination and field emergences. Phenotypic correlation between all of traits and damping- off made significant correlation (excepted to radicle fresh weight and radicle dry weight). Genotypic correlation between all of traits and damping-off made significant correlation (excepted to radicle dry weight). The genotypic correlation between hypocotyl dry weight and damping-off (after 10 days) was significant (r=-0.87, p>0.01) and between germination rate and damping-off (after 10 days) was significant (r=0.85, p>0.01). Genotypic variation of normal seedling number was 78, and genotypic variation of 10, 20, 30 days damping-off were 213, 224 and 187.

104 Budapest, Hungary, May 17th – 19th, 2004

159 - RESIDUE EFFECT OF SEED TREATMENT WITH CARBOXIN-THIRAM IN RELATION TO SOME SEED-SEEDLING PARAMETERS IN COTTON AZADDISFANI, F. & ZANGI, M.R.

Board of scientific, Cotton research Institute of IRAN, [email protected]

Seedling damping-off disease is important cotton disease in the world. Commonly, seed treatment (especially Carboxin-Thiram) control disease. Because unused seed treated, unfavorable effects of seed treatment with Carboxin-Thiram are important. In this research, Sahel cultivar seed was treated with Carboxin-Thiram. After 120 days, different characters were studied. Traits were first germination count, second germination count, third germination count, germination rate, normal seedling number, hypocotyl length, radicle length, dry and fresh weight of radicle and hypocotyl in laboratory. Results showed that CarboxinThiram decreased first germination, second germination, third germination, normal seedling number, germination rate, hypocotyl length.

160 - STUDY OF SAFFLOWER SEED MYCOFLORA IN IRAN

BAHADORANI, H. & RHIAHI, H.

Mycology Lab, Department of Biology, Shahid Beheshti University, Evin,Tehran, IRAN [email protected]

Fungal species associated with seed of 20 varieties of safllower seeds (Carthamus tinctorius) from different part of IRAN were studied. For Isolation of fungi, different methods such as Agar plate , Blotter and Freezing methods were used. For each test four hundred seeds of each varieties were tested using four replication.In total, 411 fungal strain were isolated from seed coats and emberyo .The number of fungal genera and species was 17 and 34 respectively. The maximum association was recorded for Alternaria carthami Chowdhury (35.4%), A. alternata Keissler(12%), Acromonium strictum W.Gams (1.25%), Aspergillus niger Van tiegham (11%), A. fumigatus fresen (4%), Fusarium moliniforme J.sheld (3.75%), Penicillium chrysogenum Thom (7.3%) and Ulocladium atrum Preuss (4%). In addition, species such as Acromonium strictum W.Gams, Chaetomium globosum kunze, Cladosporium herbarum preuss, C.macrocarpum preuss, C. spaerospermum Penz, Mucor racemosus Fresen, Stemphylium botrysum Wallr, Ulocaldium botrytis preuss and U.charactarum (preuss) Simmons were reported from safflower seed for the first time.Among the all varieties, maximum and minimum incidence of fungi were observed in Mahali Esfehan (10.43%) and Arac 295 (1.27%) respectively. This study attempt was made to identify mycoflora of safflower to provide information on seed health program.

161 - SEED-BORNE FUNGUS IN A TRITICUM SEED COLLECTION OF A GERMPLASM BANK BLANCO1, R., DE LA CUADRA2, C., BIELZA3, P. & TELLO1, J.C.

1Departamento de Producción Vegetal, ETSI Agrónomos, Universidad de Almería. La Cañada de San Urbano s/n, E-04120 Almería, Spain 2Centro de Recursos Fitogenéticos, INIA. Apartado 1045, 28800 Alcalá de Henares, Madrid, Spain 3Departamento de Producción Agraria. Escuela Técnica Superior de Ingeniería. Agronómica. Universidad Politécnica de Cartagena. Paseo Alfonso XIII, s/n 30203 Cartagena

The Centro de Recursos Fitogenéticos is dedicated to the conservation of plant genetic resources for food and agriculture, it is the documentation Center and manage de seed base bank of the national network in this subject and posses large active collections of cereals and legumes. The Triticum aestivum genetic resources collection in the genebank of CRF is formed by 1759 accessions with a high percentage of Spanish landraces. The multiplication and characterization are well done and the seeds have not presented any problem after a period of conservation (desiccated and cold stored) of ten years.

105 27th ISTA Congress – Seed Symposium

It is very important to have in account the seeds health of the collection, as recommended by FAO/IPGRI, for good conservation practices and to avoid pathogens extension through germplasm exchange. But in the last year a new reason reinforce this importance, because it seems that fungus exudations present in the flour for food can affect human health. For all these reasons was established a cooperation between the curator of CRF and the pathologists of the Plant Production Department at Almeria University with this subject: Screening of the accessions from the point of view of its sanitary state: detection of seed borne pathogens fungi specially dangerous for Spanish crop, i.e. Fusarium, and evaluation of fungus exudation as a sources of human problems through the flour for food. 290 accessions of Triticum aestivum and 420 of Triticum turgidum Spanish landraces were analyzed to detect presence of seed-borne pathogen fungi. One hundred seeds (ten Petri dishes with ten seeds each) were incubated under agar plate test methodology, using PDA substrate (and also KOMADA for Fusarium species) during 2-6 days at 17-23ºC. Fungi identification under microscope. As we can observe the fungi species that have a significant presence in this Triticum genetic resources collection are: Alternaria and Fusarium moniliforme in T. aestivum and Alternaria, Cladosporimu, Fusarium moniliforme and Penicillium in T. turgidum. These fungi species have been selected to compared the number of infected seeds with the percentage of germination, hundred seed weight, origin and multiplication year of the accessions, in order to look for a correlation between them. The data will be subjected to analysis of variance and correlation coefficient.

162 - ERADICATION AND EPIDEMIOLOGY OF SEED-BORNE DRECHSLERA TERES ON BARLEY CARMONA 1, M.A., BARRETO 1,2, D.E., MOSCHINI3, R. & REIS 4, E.M.

1Fitopatología. Facultad de Agronomía. Universidad de Buenos Aires. Av. San Martín 4453 (1417), Capital Federal, Argentina, [email protected]) 1y2 IMYZA, INTA, Castelar, Buenos Aires, Argentina 3Instituto de Clima y Agua , INTA, CC Nº 25 (1712) Castelar, Argentina. 4Fitopatología. Faculdade de Agronomía e Medicina Veterinaria Universidade de Passo Fundo. C. P. 566, 99001-970, Passo Fundo R. S. Brasil.

Net blotch, caused by Drechslera teres (Sacc.) Shoem., is an important disease of barley in Argentina. Seed is an efficient mean for both short and long distance dissemination. To prevent the introduction of the pathogen into the field, seed treatment has been recommended. However, the efficiency of fungicides is about 50 to 70%. More adequate fungicides need to be tested. The objectives of this research were to evaluate fungicides for eradicating D. teres in the seed and to study disease epidemiology in naturally infected and eradicated seeds. In vitro, three fungicides were evaluated: iminoctadine, guazatine and thiram + iprodione at 150, 200, 250 and 300 g or ml /100 kg of seed. Seeds of Q. Pampa with 68.75% of D. teres incidence was used in the tests. The epidemiology study, under field conditions, including two plots of 80 m2, separated by 20 m. One plot was sown with eradicated seed (iprodione + thiram, 300 g / 100 kg) and the second plot was the untreated check. Crop growth and disease observations were recorded at 7 days intervals. The incidence of diseased plumules and leaves, severity, airborne spores, yield components and seed re-infection, were assessed. In vitro test indicated that the three fungicides eliminated D. teres at the dosage of 300 g or ml. Under field conditions, the eradication was not obtained, but the disease was significantly less severe in the treated plot and the disease severity and did not reach the alarm threshold for foliar chemical control.

106 Budapest, Hungary, May 17th – 19th, 2004

163 - HARVEST AND STORAGE OF THE COMMON AND HARD WHEAT SEEDS HEALTHINESS. CARNEIRO1, . L.M.T.A., BIAGI2, J.D., FREITAS3, J.G., CARNEIRO4, M.C. & BOLPETTI5, A.N.

1UNICAMP, Campinas State University, Brazil, [email protected] 2UNICAMP, [email protected] 3Campinas Agronomy Institute, Brazil 4,5UNICAMP

The contamination with molds during the storage, is a factor that, interacting with temperature and humidity, may increase the deterioration of stored seeds. The objective of the work was to evaluate the influence of the time of harvest and the storage in the seed healthiness of common and hard wheat. The experiments were installed with 2 genotypes of common wheat (Triticum aestivum L), IAC-24, harvested with 30.0, 21.2 and 12.2% of moisture content and IAC-289, harvested with 35.0, 23.4 and 12.5% of moisture content; e 1 genotype of hard wheat (Triticum durum L.) IAC-1003, harvested with 31.6, 22.2 and 11.7% of moisture content. The seeds of 1st and 2sd harvest were dried at 40, 60 and 80°C until they reached 13.0% of moisture content. Dried seeds were then stored in polyethylene packets for a period of 0, 2, 4, 6 and 8 months at 20±2°C, and the seed healthiness were evaluated using the blotter test at IAC’s Plant Pathology Laboratory and the water activity was determined during the storage. With the early harvest, it had a reduction of the incidence of some molds of field, as Alternaria alternata, Bipolaris sorokiniane and Fusarium spp, and a higher incidence of molds storage, mainly the Penicillium spp. The seeds harvested with higher moisture content showed a higher water activity.

164 - POTENTIALITY OF THE WATER RESTRICTION TECHNIQUE IN HEALTH TESTING AND IN STUDIES ON THE INTERACTION OF BIPOLARIS SOROKINIANA AND WHEAT SEEDS CELANO, M.M1; MACHADO, J.C1.; JACCOUD FILHO, D.S2 & GUIMARÃES, R.M1

1Universidade Federal de Lavras,MG, Brazil, CEP 37200-000, [email protected] 2Universidade Estadual de Ponta Grossa, PR, CEP 84010-330

Results of previous investigation indicated that water restriction of blotter substrate produced by mannitol and NaCl at potential of -0.8 to -1.1 MPa proved to be efficient to reduce seed germination of wheat seeds during incubation period of 7 days without affecting fungal development. From a comparison between both freezing and water restriction techniques by examining sixty seed samples it was shown that mean incidence of Bipolaris sorokiniana was similar in both cases and incidence of Pyricularia grisea, Aspergillus ochraceus, Cladosporium cladosporioides and Epicoccum purpuracens was higher in water restriction condition. Mean incidence of Fusarium graminearum and Drechslera tritici repentis was lower at higher osmotic potential of the blotter substrate. In the study on the interaction of Bipolaris sorokiniana and wheat seeds, the use of water restriction technique was of extreme help in grouping seeds with different inoculum potential of that fungus. From the sowing of those different groups of seeds it was possible to evaluate the effects of that fungus on the performance of the seeds by means of germination and vigour tests and measurement of plant size and weight. Seeds with higher inoculum potential of B.sorokiniana presented the lowest values of all variables considered in the evaluation of this trial.

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165 - PLANTS WITH PROSPECTS TO BE USED IN AGRICULTURAL CROPS PROTECTION AGAINST DISEASES CHAKAEVA1, A.SH., KOTOVA2, V.V. & CHERMENSKAYA2, T.D.

1Kyrgyz Research Institute for Livestock, Veterinary and Pastures, [email protected] 2All-Russian Institute of Plant Protection, [email protected])

Search for new biologically active compounds of plant origin is a promising direction of the development of ecologically safe means of plant protection from a complex of hazardous organisms. The study and application of biologically active plant substances in phytopathogen control take on special significance nowadays, what is connected with the ecological importance of plant protection and the tasks of environmental protection. But many aspects of this problem remain insufficiently studied. We have carried out the search and study of new active compounds of plant origin having fungicide and growth-stimulating activity. Over the period of 2002-2003, we have carried out the screening of more than 50 plant extracts and estimated the effect that the biologically active substances contained in them had on phytopathogenic fungi. The laboratory screening of showed that some preparations of this range displayed good results in restricting phytopathogenic fungi colonies growth on nutrient mediums, suppressing their spore genesis, changing the colonies texture and colour. Plants of the family Ranunculaceae (Akonit soongaricum Stapf. and Adonis parviflora Fisch.) deserve special attention. The interest to these plants is rooted in ancient times. For example, the Greek name was given to aconite by Dioscorides, “akoniton” meaning “poison”, that is the plant of which a poisonous substance was obtained to kill beasts of prey. Later it was found out that this plant contained the alkaloid aconitine and its derivatives (Gorelova, 1986). It is also known about the application of aconite in plant protection. According to V.F. Kovtunenko e.a. (1972), the extract of aconite leaves was successfully used to control the tick Tetranychys urticae Koch, the green apple louse, the raspberry weevil and some leaf- eating insects. It has also been determined that the extracts have fungicidal effect on the powdery mildew of apple-trees and other crops. According to literature data (Poshkurlat, 2000; Komissarenko e.a., 1975) adonis contains a number of biologically active substances belonging to different polyphenol compound (flavonoids, glycosides, coumarins, etc.), as well as water-soluble polysacharides, including adonitoxin. All these substances can act as biogenic elicitors – substances that can induce defense reactions in plants and increase their resistance to diseases (Tyuterev, 2002). In the series of our experiments in vitro 1% solutions of ethanolic extracts of these plants can inhibit the growth of phytopathogenic fungi: Fusarium oxysporum (alfalfa isolate), Verticillium albo-atrum (alfalfa isolate), Fusarium nivale, F. solani (isolates of fodder plants) and Colletotrichum lupini when they are cultivated on nutrient mediums. So, when the aconite extract was introduced in the nutrient medium (KDA), the colony diameter of the fungi decreased after 9 days of their cultivation on nutrient mediums as follows: V. albo- atrum - 3 times, C. lupini – 3 times, F. oxysporum и F. nivale. 2 times in comparison with the control variant (Fig. 1). The aconite extract suppressed the spore genesis of the fungus V. albo-atrum и C. gloeosporioides sharply. The productivity of spore formation (the number of the fungus conidia) on the nutrient mediums treated with the aconite extract reduced 9 times in V. albo-atrum and 5 times in C. lupini in comparison with the control test. The microscopic analysis revealed different morphological changes, anomalies in the character of fungi mycelium growth, the development of mainly substrate weakly branching mycelium, its vacuolization; in some hyphas the aggregation of the cytoplasm and the destruction of hyphas was observed. The data show the high fungicide activity of aconite extracts. Some regularities of phytopathogenic fungi inhibiting in vitro were also established when adonis was used. It was recorded that the growth of the fungus strains tested was reduced by 8-30%. In vegetation experiments against a severe infection background with the introduction of the infectious material of F. oxysporum (10 g per kg of soil) and sowing alfalfa seeds treated with 1% solution of the ethanolic extract of adonis (the seeds had been soaked for 15 minutes) a protective effect and an increase in the resistance to fusarial root rot was determined. The infection of sprouts and shoots by F. oxysporum and, consequently, the

108 Budapest, Hungary, May 17th – 19th, 2004 destruction of plants in the variants with treated seeds declined from 55% (in the control variant) to 12.5 % (Fig. 2). The high stimulating effect of the adonis extract was established also after the treatment of sainfoin and spring wheat seeds. Thus, an increase in the laboratory germination capacity of sainfoin from 60 to 80 % was recorded, the sprouts mass grew 1.8 – 2 times; the seed germination energy of spring wheat rose by 30%. In field conditions, the treatment of roses, apple-trees and potatoes with the extracts of adonis and aconite performed twice (in June and July), under the conditions of the epiphytous disease development, promoted the reduction of the development of hazardous diseases on these crops. The data presented on the diagrams (Figures 3 and 4) evidence the high protective effect of the preparations against Sphaerotheca pannosa Fr. Zev. Var. Rosae Woron. and Venturia inaequalis (CKe) Wint. The biological effectiveness of the adonis extract was 60% against Sphaerotheca pannosa Fr. Zev. Var. Rosae Woron., and 61% against Venturia inaequalis (CKe) Wint. The biological effectiveness of the aconite extract was 85.6% against Sphaerotheca pannosa Fr. Zev. Var. Rosae Woron. and 71.7% against Venturia inaequalis (CKe) Wint. The effectiveness of the extracts in the control of potato phytophthora rot was insignificant and did not exceed 37.5%. So, Adonis and aconite may be regarded as promising plants to be used in the development of ecologically clean means of protecting plants from diseases.

Bibliography

1. A.P. Gorelova. Introduction of aconites in the subarctic zone.- The Academy of Sciences of the USSR, the Polar-Alpine Botanical Garden, Apatity, 1986.- p. 111-116. 2. V.F. Kovtunenko, T.D. Budarina, T.I. Plekhanova, D.A. Muravyova. On the incecticide and fungicide properties of Aconitum. In: Plant Protection from Pests and Diseases. Moscow, the GBG of the Academy of Sciences of the USSR, 1972. – p. 148-150. 3. N.F. Komissarenko, V.Ya. Yatsuk. The chemical compounds of the overground part of Adonis – Rastitelnye Resursy, 1975.- Vol. 11.- Issue 4.- p. 515-517. 4. A.P. Poshkurlat. The genus Adonis L.- Moscow: Nauka – Interperiodika, 200, 1982.- p. 171-193.

166 - NATIVE TREE SEEDBORNE FUNGI FROM THE BRAZILIAN ATLANTIC FOREST. DOS SANTOS, A.F., DE S. MEDEIROS, A.C. & DE SOUSA, V.

Embrapa Florestas – National Forest Research Center of EMBRAPA, Cx. P. 319, 83411-000, Colombo, PR – BRAZIL, [email protected]

The purpose of the present work was to investigate and identify fungi on seeds of five native trees. Two methods (standard blotter and agar plate test) were used to evaluate the incidence of fungi on seeds. Twenty three fungi genera were identified: Aspergillus sp., Penicillium sp, Pestalotia sp., Helminthosporium sp., Trichoderma sp., Chaetomium sp., Monilia sp., Alternaria sp., Ulocladium sp., associated with “pau-cigarra” (Senna multijuga, Leguminosae-Fabaceae) seeds; Fusarium sp., Alternaria sp., Pestalotia sp., Penicillium sp., Aspergillus sp., Trichoderma sp., Chaetomium sp.,Nigrospora sp., Geotrichum sp. and Mucor sp. associated with “aroeira-vermelha” (Schinus terebinthifolius, Anacardiaceae) seeds; Fusarium sp., Phomopsis sp., Colletotrichum sp., Cladosporium sp., Alternaria sp., Aspergillus sp., Pestalotia sp., Monilia sp.,Trichoderma sp., Penicillium sp. and Geotrichum sp associated with “angico” (Piptadenia paniculata, Leguminosae-Fabaceae) seeds; Fusarium sp., Cladosporium sp., Alternaria sp., Septoria sp., Botryodiplodia sp. and Colletotrichum sp. associated with “vacum” (Allophylus edulis, Sapindaceae) seeds; and Phomopsis sp., Phoma sp., Cladosporium sp., Fusarium sp., Curvularia sp., Peronospora sp., Pestalotia sp., Penicillium sp., Aspergillus sp., Trichoderma sp., Rhizopus sp., Chaetomium sp., Ascochyta sp. and Stilbum sp. associated with “cedro” (Cedrela fissilis Vell., Meliaceae). It was concluded that the best management is need in order to reduce seed contamination of these native species.

109 27th ISTA Congress – Seed Symposium

167 - CONTROL OF SOYBEAN SEED-BORNE PATHOGENS BY FOLIAR FUNGICIDE APPLICATIONS IN ARGENTINA GALLY, M.1, CARMONA, M.1, GRIJALBA, P.1, BARRETO, D.1Y2, SUGIA, V.1 & ARRUABARRENA, N.1

1Facultad de Agronomía, Universidad de Buenos Aires, Argentina. [email protected] 2IMYZA, INTA, Castelar, Buenos Aires, Argentina

Pampeana Central Region is the main important soybean (Glycine max) producing area of Argentina. Combination of no tillage and soybean monoculture has caused great increment of damages caused by diseases last years. The prevalent late season diseases of soybean in the region are: brown spot (Septoria glycines), Cercospora blight (C. kikuchii), anthracnose (Colletotrichum truncatum, Glomerella glycines) and pod and stem blight (Diaporthe phaseolorum var. sojae). Some of these pathogens are often found infecting harvested seeds. The objectives of this study were to evaluate the incidence of seed-borne pathogens and the efficiency of foliar fungicides to reduce seed infection. Trials were conducted through 2001 to 2003 growing seasons in no-tilled monoculture commercial soybean crops. Treatments were arranged in a complete random block design. Triazoles, benzimidazoles and strobirulins fungicides, pure or mixed, were applied once in reproductive soybean stages R3 or R5. Seed health testing was conducted using the blotter paper test. Most frequent pathogens in seeds were: C. kikuchii, Phomopsis sojae., and different species of Fusarium. All fungicide treatments significantly controlled only C. kikuchii. Yield was significantly higher in treated plots. No difference was detected between the time of fungicide application. It was concluded that one foliar application of fungicide in R3 or R5 did not reduce seed infection by some important seed-borne pathogens under the environment conditions of the Pampeana Region.

168 - EFFECTS OF SEED INFECTION LEVEL BY ASCOCHYTA PINODES ON BLIGHT DEVELOPMENT AND SEED YIELD OF FIELD PEA UNDER FIELD CONDITIONS GORFU1, DEREJE & SANGCHOTE2, SOMSIRI

1Holetta Agricultural Research Center, P.O.Box 2003, Addis Ababa, Ethiopia., [email protected] 2Kasetsart University, Department of Plant Pathology, P.O.Box 1090, Bangkok 10903, Thailand., [email protected]

The development of Ascochyta blight [Ascochyta pinodes (teleomorph = Mycosphaerella pinodes)] and seed yield of field pea (Pisum sativum) were studied using 0, 10 and 20% seed infection levels of three varieties under field conditions at Denbi and Holetta, Ethiopia, in 2002. Seed infection significantly influenced the emergence percentage (EP) being 81.7, 79.6 and 73.9% at Denbi and 87.7, 83.0 and 81.5% at Holetta for 0, 10 and 20% seed infection respectively. Ascochyta infection started as foot rot, minute flecks on stems and rarely on leaves at about five weeks after sowing (WAS) and the incidence accorded seed infection up to about seven WAS. Then a sharp increase of incidence to over 95% within a week at different times at both locations subverted the effect of seed infection. After establishment, the blight progress was not so different for seed infection levels and varieties. A steady increase of blight was recorded until mid September and then faster only at Denbi. Seed yield was significantly influenced by seed infection with a mean of 1.03, 0.80 and 0.71 t/ha at Denbi and 2.23, 2.16 and 1.98 t/ha at Holetta for 0, 10 and 20% seed infection respectively. Seed infection by A. pinodes up to 10% showed no significant yield reduction in the subsequent crop.

110 Budapest, Hungary, May 17th – 19th, 2004

169 - ROLE OF BIOCONTROL AGENTS IN THE SUPPRESSION OF SEED BORNE PATHOGENIC MYCOFLORA AND THEIR EFFECT ON SEED QUALITY IN SORGHUM [SORGHUM BICOLOR (L.) MOENCH] INDIRA, S., MUTHUSUBRAMANIAN, V., TONAPI, V.A., & SEETHARAMA, N.

National Research Centre for Sorghum, Rajendranagar, Hyderabad, Andhra Pradesh 500 030, India, [email protected]

The scope of biocontrol agents in the control of sorghum grain molds was investigated both invitro as well invivo. Their utility in enhancing the germination and seedling vigour of mold infected seeds were assessed. The biocontrol agents, Trichoderma viride, T. harzianum, T. hamatum, T. koeningii and Pseudomonas fluorescens (Isolates collected from the state of Tamil Nadu, India) were efficient in checking the growth of mold pathogens when grown in Potato Dextrose Agar (PDA) as dual cultures. The bacterial bioagent P. fluorescens was most efficient in checking the growth of all the test pathogens viz., Fusarium moniliforme, Curvularia lunata and Alternaria alternata (66.8, 68.5 and 64.9 % inhibition of colony growth respectively) followed by other bioagents for their antagonistic effect. The in vivo effectiveness of the bioagents got enhanced when combined with other components like host-resistance, cultural and chemical control strategies, paving way for a possible Integrated Disease Management. The percent incidence and severity of molds were lowest (19.44 and 13.5 respectively) with the treatment T6 (seeds harvested at physiological maturity + propiconazole @ 0.02 % + P. fluorescens @ 1× 108 cfu ml-1) that recorded maximum 100 seed weight (2.905 g), followed by 2.871g in in T2 (seeds harvested at physiological maturity + P. fluorescens). The effect of bioagents in enhancing the germination and seedling vigour was most promising. Of all the treatments, P. fluorescens resulted in maximum germination (88 %) and seedling vigour (2635.60 vigour index), followed by treatment with T. viride (86.6 % and 2478.47 respectively).

170 - HEALTH QUALITY OF SEEDS IMPORTED BY THE STATE OF PARANÁ JACCOUD-FILHO1, D.S., HIAR, C.P., COLTURATO, A.B., BOBATO, E., PASSINI, F.B. ZAMPIER, I. & MELLO, R.P.

Universidade Estadual de Ponta Grossa, UEPG. Av. Carlos Cavalcanti 4748, Campus Uvaranas, Bloco F, CEP 84030-900, Ponta Grossa-PR, BRASIL, [email protected]

The state of Paraná stands out nationaly for its developed and highly technical agriculture. However, due to the limited production of seeds in Brasil, especially that of vegetables, flowers and spices, Paraná needs to import a significant amount of seeds from many countries. The objective of this project was to identify and measure the level of incidence of fungi present in the imported seeds, as well as to evaluate the potential risk of contamination of such seeds for the agriculture of the state of Paraná. The method used was the Blotter Test Method, with the seeds being incubated for a period of 12h day light / 12h dark, at a temperature of 22ºC for seven days. Afterwards, the seeds were observed in steroscopic and commom optical microscopes for the identification of the fungi present. 251 batches of seeds were analysed, of which 69% were of vegetables, 31% were flowers, spices and medicinal plants, from EUA (32%), Denmark (31%), Italy (18%), Chile (6%), France (5%), Holland (5%), Japan (2%) and Germany (1%). The fungi observed with more frequency were: Alternaria alternata, Fusarium spp., Alternaria brassicicola, Curvularia sp., Phoma sp., Drechslera sp., Bipolaris sp., Lepthosphaerulina sp. and Alternaria zinniae. Among the countries that export seed to Paraná, the higher frequencies of fungi incidence were observed in seed batches from France (71%), Italy (71%), Holland (71%), Chile (65%), Denmark (54%), EUA (46%) and Japan (14%) respectively. In general, the seed batches presented a satisfactory health level, probably because the majority of the batches had been previously treated with fungicides. The elaboration of a catalogue with the fungi identified in the seeds of the different species is in its final phase.

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171 - DEVELOPMENT AND USE OF ALTERNATIVE METHODS OF SEED TREATMENT AGAINST SEED-BORNE PATHOGENS IN ORGANIC FARMING JAHN, M.1 & KOCH, E.2

Federal Biological Research Centre for Agriculture and Forestry 1Institute for Integrated Plant Protection, Stahnsdorfer Damm 81, D-14532 Kleinmachnow, [email protected] 2Institute for Biological Control, Heinrichstraße 243, D-64287 Darmstadt

In the future, conventionally produced seed will not be allowed for organic farming. A deadline for deviations is set at 1 January 2004. Seed-borne diseases tend to increase and develop into a problem without adequate control. According to the principles of organic farming to cut out synthetic plant protection products effective non-chemical seed treatment methods are necessary for the control of seed-borne diseases. Physical as well as biological measures can be taken into account. Among physical methods, the „classical“ methods of thermotherapy, partly combined with moisture (hot water, hot steam, hot air), but also modern methods like electron or microwave treatment seem to be possible. Among biological methods, applications of plant strengthening products (e.g. botanicals, microbial products) and biological plant protection products (e.g. botanicals, antagonists) are a possibility. In the last decade, a lot of work has been done to develop several measures or to adapt such methods to different crops. Hot water treatment, hot air treatment, electron treatment, treatments with plant extracts, and treatments with microbial products have been examined for the use in cereals and vegetables. The aim is to introduce this methods in agricultural and horticultural practice. Important cereal and vegetable crops (wheat, rye, barley, carrot, cabbage, celery, parsley, lamb´s lettuce) with their most important seed-borne pathogens (Tilletia caries, Ustilago spp., Fusarium spp., Alternaria spp., Phoma spp., Septoria spp., Peronospora valerianellae, Xanthomonas campestris) are included. It can be established that the control of most seed-borne pathogens on the plants is possible by one or more of the mentioned methods. Altogether, it is now the aim to provide a rank of alternative measures to select the most suitable method for each special situation.

172 - MYCOFLORA AND POLYPHENOLS OF BAMBARA GROUNDNUT KOLA, M.E.1, AVELING, T.A.S.1, REGNIER, T.J.C.1 & HALL, A.N.2

1Department of Microbiology and Plant Pathology, Forestry and Agricultural Biotechnology Institute (FABI) 2Laboratory for Microscopy and Microanalysis, University of Pretoria, Pretoria 0002, South Africa

Bambara groundnut (Vigna subterranea (L.) Verdc.) is an important legume crop which is rich in protein content and is grown and consumed by small-scale farmers. In this study the percentage mycoflora infecting seeds of variously coloured Bambara groundnut varieties was determined. Seeds of six different coloured varieties were surface sterilised, plated on agar media, incubated for seven days and the fungi were identified and percentage infection determined. Dark coloured seeds were less infected by seed mycoflora than light coloured seeds. To determine whether seed coat surface topography played a role in infection, seeds were dissected in two, placed on stubs, coated with gold and examined with the scanning electron microscope (SEM). The concentration of phenylpropanoids in the seed and the localisation of the main phenolic compounds were also determined. Seed coat sections (10 µm thickness) were cut with a Cryostat microtome and mounted in glycerine or vanillin-HCl and stained with Neu reagent. The sections were viewed under normal and/or UV (360 nm) and blue (420 nm) light illumination using an epifluorescence microscope. SEM indicated that there were no distinct morphological differences among the topography of the different coloured seed coats. Dark coloured seed coats stained with vanillin-HCl were darkly pigmented red showing the presence of proanthocyanidins in the epidermis and parenchymatous layer. Dark coloured seeds emitted a strong blue fluorescence under UV illumination indicating the presence of large quantities of polyphenolic compounds, which may play a role in suppressing the infection of these seeds by mycoflora.

112 Budapest, Hungary, May 17th – 19th, 2004

173 - ULTRASTRUCTURAL DAMAGE AND GERMINATION REDUCTION IN COWPEA SEEDS DUE TO FUMONISIN B1 KRITZINGER, Q. & AVELING, T.A.S.

Department of Microbiology and Plant Pathology, Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria 0002, South Africa

Cowpea [Vigna unguiculata (L.) Walp] seed plays an important role in the nutrition of many subsistence farmers and rural communities. The seeds are prone to fungal infestation and mycotoxin contamination during sub-optimal storage conditions. Fumonisin B1 (FB1), primarily produced by Fusarium verticillioides and F. proliferatum, has been detected in cowpea seeds. Surface-disinfected seeds were imbibed for 10 h in 50 ml sterile distilled water amended with FB1 to yield final concentrations of 10, 25, 50, 100 ppm. Slow imbibed seeds (placed in moist paper towels) were incubated at 25˚C for 10 h (positive control). Seeds placed in sterile distilled water for the same period of time served as the negative control. Each treatment was replicated three times. Percentage germination was determined according to the International Seed Testing Association (ISTA) rules. Radicle and plumule length was measured after 8 days. Parts of the embryonic axes and cotyledon tissues were removed and prepared for transmission electron microscopy. All the toxin concentrations significantly decreased seed germination. Radicle elongation was inhibited by 10, 25 and 50 ppm concentrations whereas no significant reduction in plumule length was noted. FB1 treated embryonic tissues indicated compaction of the protoplasm and separation of the plasmalemma from the cell wall. Lipid bodies accumulated, which seemed to be lining the cell wall. This was also found in treated cotyledon tissues. It appeared that the lipid bodies were structurally disrupted. This is the first study to demonstrate the phytotoxic effects of FB1 on cowpea seeds.

174 - INTEGRATED RESEARCH PROGRAMS ON SEED PATHOLOGY AND RELATED FIELDS AT THE FEDERAL UNIVERSITY OF LAVRAS-MG, BRAZIL: A SUCCESSFUL EXPERIENCE MACHADO, J.C., VIEIRA, M.G.G.C., GUIMARÃES, R.M., VON PINHO, E.V.R, CARVALHO, M.L.M OLIVEIRA, J.A. & VIEIRA, A.R.

Federal University of Lavras (UFLA), MG, Brazil,- CEP 37200-000, [email protected]

The integrated approach of seed pathology, seed production and technology is a goal that has been pursued at the Federal University of Lavras for many years in the attempt to tackle problems on a global view looking at maximization of resources. Faculty members of the University involved in the seed production chain come together in projects with participation of students of different levels. Graduate dissertations and thesis are included in those projects and developed under the supervision of multi-discipline Committees. The definition of topics for investigation is based on demands collected to seed producers and users. Successful examples of integrated programs are the studies focusing on application of water restriction/ osmo-priming technology in seed health testing and in the process of infecting seeds with pathogens. Effects of microorganisms in seed physiology testing - tetrazolium, electrical conductivity and aging have been also investigated and the results show the need to pay more attention to such interactions. Interference of microorganisms in results of genetic purity testing has been also demonstrated in some cases. Studies on interaction between seed and microorganisms during storage reveal that, for many cases, inoculum viability of field fungi declines during storage, then leading seeds to express germination at their potential. In studies on chemical seed treatment performance of the treatment is largely influenced by the physiological conditions of the seeds.

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175 - EVALUATION ON THE INDUCTION OF PATHOGENESIS-RELATED PROTEINS BY METHYLOBACTERIUM SPP PPFMS-OS-07 AND ITS EFFECT ON RICE CHALLENGED WITH RHIZOCTONIA SOLANI MADHAIYAN1, MUNUSAMY, SENTHILKUMAR2, MURUGAIYAN, SESHADRI1, SUNDARAM, SUNDARAM2, SUBBIAH & SA1, TONGMIN

1Dept. of Agricultural Chemistry, Chungbuk National University, Cheongju, Chungbuk, 361-763, Korea, Tel.: 82-43-261-2561; Fax.: 82-43-271-5921, [email protected] 2Dept of Agricultural Microbiology, Tamilnadu Agricultural University, Coimbatore 641 003, Tamilnadu, India

Pink-pigmented facultative methylotrophic (PPFM) bacteria belonging to Methylobacterium spp are persistence colonizers of all plant leaf surfaces and transmit through seeds. They utilize methanol as preferred carbon source and can exert slow growth on multicarbon compounds. They have been shown to stimulate seed germination and plant development. In the present study, we have studied the impact of an Indian isolate Methylobacterium sp. (PPFMs-Os-07) on rice (var. Co47) seed germination and plant growth. Significant differences were observed when paddy seeds were treated PPFM and on foliar sprays. Significant control of sheath blight was also observed when plants were inoculated with Rhizoctonia solani (TNAU-01) under pot culture conditions. Rice plants sprayed with Methylobacterium spp strain PPFMs-Os-07 showed increased activity of all enzymes and phenolic content on day one after application. Maximum activity of phenylalanine ammonia lyase and peroxidase was observed on fourth day and β-1,3-glucanase and chitinase on fifth day after treatment.

176 - INTRODUCTION OF SEED BORNE ENDOPHYTIC FUNGI FOR ENHANCEMENT OF STRESS TOLERANCE IN TALL FESCUE NEKOUEI 1, M. KHAYYAM, MIRLOHI 2, AGHA.FAKHR & SHAHAB3, M. NADERI

1Department of Biotechnology, Isfahan Research Center of Agriculture and Natural Resources, Iran, Isfahan, P. O. Box: 81785-199 2Department of Agronomy and Plant Breeding, College of Agriculture, Isfahan University of Technology, Iran 3Agriculture Biotechnology Research Institute of Iran, Iran, Karaj, P. O. Box: 31535-1897

Tall fescue is a popular pasture grass grown in many countries. A systematic endophytic fungus, Acremonium coenophialum, lives in a symbiotic association within tall fescue and may impart superior competitiveness to the plant through increased resistance to pests, tolerance to drought and improvements in other agronomic traits. The assessment of the infection status and viability of endophytic fungi would open the possibility of identifying potentially novel endophyte strains for enhancement of stress tolerance in grasses. In this study several accessions of tall fescue were collected from various regions of Iran, identified and evaluated for the presence of endophyte. The accessions were mainly distributed in the northern and western part of the country with relatively more precipitation. Detection of endophytic fungi in tall fescue seeds showed that 84.2% of the accessions were infected with endophyte at infection rates of 20 to 95%. The results of the endophytic fungi detection in greenhouse-grown and field-grown tall fescue seedlings indicated that viable fungal endophyte occurred in 73.3% of total tall fescue accessions evaluated. The in vitro isolation and culture of endophyte confirmed the result obtained from greenhouse and field experiments.

114 Budapest, Hungary, May 17th – 19th, 2004

177 - EVALUATION OF FIVE ESSENTIAL OILS FROM AROMATIC PLANTS OF CAMEROON FOR CONTROLLING SEED-BORNE BACTERIA OF RICE (ORYZA SATIVA L.) NGUEFACK1,3,*, J., SOMDA1,2, I., MORTENSEN1, C.N. & AMVAM ZOLLO3, P.H.

1Danish Government Institute of Seed Pathology for Developing Countries (DGISP), Thorvaldsensvej 57, DK-1871 Frederiksberg C., Denmark. 2Rural Development Institute, Polytechnic University of Bobo-Dioulasso, Burkina Faso. 3Department of Biochemistry, University of Yaounde-I, Box 812 Yaounde Cameroon. *Corresponding author address Department of Biochemistry, University of Yaoundé-I, Box 812 Yaoundé Cameroon. E-mail: [email protected] or [email protected]; Tel.: +237 2312243

Five essential oils (EO) extracted from Cymbopogon citratus, Monodora myristica, Ocimum gratissimum, Thymus vulgaris and Zingiber officinale were investigated for their inhibitory effect against five seed-borne bacteria, Acidovorax avenae subsp. avenae, Burkloderia glumae, Burkholderia plantarii, Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. orizicola. Different degrees of bacterial growth inhibition by the EO at 1:10 dilution (v/v) were recorded in term of inhibition zone using the agar diffusion method. The EO from O. gratissimum and T. vulgaris were the most effective giving against the five bacteria tested mean inhibition zones of 14 mm and 10 mm diameter, respectively. Moderate activity was observed for the EO from C. citratus and Z. officinale with respectively, mean inhibition zones of 7 mm and 2 mm diameter, while the EO from M. myristica was less inhibitory. X. oryzae pv. oryzicola and X. oryzae pv. oryzae were the most and the least sensitive strains regardless the EO. These effects against seed-borne bacteria indicated the possible ability of each essential oil as a seed protectant. The potential use of the two most effective EO extracted from O. gratissimum and T. vulgaris, was investigated for their ability to control seed to seedling transmission of A. avenae subsp. avenae and the seed-borne infection of X. oryzae in naturally infected seeds using the cassette holder method and liquid assay, respectively. Four rice cultivars were tested and the EO applied as slurry significantly reduced the seed transmission of A. avenae subsp. avenae by 60% to 80% from seeds with infection of 32%. The vigour of rice seedlings raised from the treated seeds was better compared to that of seedlings from the untreated seeds. Although we failed to detect Xanthomonads among the yellow bacteria on mXOS medium, the treatment with EO also reduced the total cells counts of bacteria at a range of 92%-97%. Both EO also increased the germination capacity of the treated seeds with 13% to 23%. On-farm evaluation need to be carry out.

178 - SEED HEALTH STATUS OF DIFFERENT COTTON VARIETIES RAJESWARI, B., MEENA KUMARI, K.V.S. & MURALIMOHAN REDDY, B.

National Seed Project, ANG Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, India, [email protected]

Seeds of twenty five varieties of cotton were subjected to seed health and germination by blotter method and germination paper towel method, respectively. Fuzzy seeds recorded more percentage of fungal flora (6.4%) as compared to delinted seeds (0.1%). The predominant seed borne mycoflora in fuzzy seeds (Aspergillus flavus, Alternaria macrospora and Fusarium moniliforme) and in delinted seeds (Aspergillus flavus and Fusarium sp.) were recorded under laboratory conditions. Fuzzy seeds recorded low per cent germination (45- 60%) as compared to delinted seeds (60-80%). Farmers are advised to adopt use of delinted cotton seeds to reduce mycoflora and increase germination.

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179 - SEED TREATMENTS FOR ORGANIC VEGETABLE PRODUCTION SCHMITT, ANNEGRET & KOCH, ECKHARD

Federal Biological Research Centre for Agriculture and Forestry (BBA), Institute for Biological Control, Heinrichstr. 243, 64287 Darmstadt, Germany, [email protected], fax: +49 6151 407290

Due to the difficulty in organic farming of producing pathogen free seeds, and the lack of simple, effective non-chemical methods for seed sanitation, a substantial part of the seed used by European organic vegetable growers is still derived from conventional production. This will be strongly restricted after the year 2003 (EU Council regulation 2092/91). In March 2003, an EU-project "Seed Treatments for Organic Vegetable Production" (QLRT- 2002-02239; STOVE) was initiated. The project is aimed at improving currently available, non-chemical methods for control of seed-borne vegetable pathogens and to develop new methods which are acceptable to organic farming.

The participants are: - Federal Biological Research Centre for Agriculture and Forestry (BBA), Germany (M. Jahn, E. Koch, C. Kromphardt, A. Schmitt) - Plant Research International (PRI), Wageningen, Netherlands (S. Groot, J. v.d. Wolf) - Swedish University of Agricultural Sciences (SLU), Uppsala, Schweden (G. Forsberg, B. Gerhardson) - University of Gothenburg, Sweden (T. Amein, S. Wright) - Findus R & D AB, Bjuv, Sweden (R. Stegmark, M. Wikström) - Nunhems Zaden BV, Haelen, Netherlands (J. v.d. Berg) - Nunhems Zaden (Hild), Marbach, Germany (M. Mistele, S. Werner) - University of Turin, (Agrinnova) Italy (M. Gullino, F. Tinivella) - Horticulture Research International (HRI), Wellesbourne, Great Britain (S. Roberts)

Together with three physical methods (hot water, hot air and electron treatment), micro- organisms and other agents of natural origin acceptable to organic farming will be included in the project. Initially, the three physical methods are being adapted for different vegetable species (e.g. carrot, parsley, cabbage, lamb`s lettuce, basil, bean) and their respective seed- borne pathogens (e.g. A. dauci, S. petroselini, X. campestris, P. valerianellae, Fusarium spp., C. lindemuthianum). In parallel, potential alternative seed treatments (micro-organisms, plant extracts, inducers of resistance) are also being tested. The efficacy of the methods will be compared in glasshouse and field trials, and selected combinations will be evaluated. Special regard will be placed on physiological factors determining the sensitivity of seeds towards the physical methods. The results of the first experiments will be presented. (See http://www.stove-project.net/ for further information).

180 - SEEDBORNE AND QUARANTINE SIGNIFICANT FUNGAL PATHOGENS OF SORGHUM, PEARL MILLET, CHICKPEA, PIGEONPEA AND GROUNDNUT THAKUR, R.P. & GIRISH, A.G.

International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru 502 324, Andhra Pradesh, India; [email protected]

The ICRISAT gene bank holds 104600 germplasm accessions of its five research mandate crops (sorghum, pearl millet, chickpea, pigeonpea, and groundnut) for their utilization in genetic improvement of these crops. The Plant Quarantine Laboratory at ICRISAT facilitates the exchange of seed material of the germplasm accessions and breeding lines following the standard seed health testing procedures and strictly adhering to the plant quarantine regulations of Government of India. During 1989-2003, seed health testing of 411760 million seed samples for export to 156 countries, 39 seedborne fungal species were recorded: 16 on sorghum; 9 on pearl millet, 6 on chickpea, 10 on pigeonpea and 8 on groundnut. The highest infected samples were of pigeonpea (6.2%) and the lowest of pearl millet (1.2%). Of these, 24 fungal species belonging to 18 genera, including Fusarium (wilt),

116 Budapest, Hungary, May 17th – 19th, 2004

Peronosclersopora and Sclerospora (downy mildew), Sporisorium (smut), Claviceps (ergot), Ascochyta (blight), Colletotrichum (anthracnose), and Macrophomina (root rot) were of plant quarantine importance. Some of the infected seed samples were treated with systemic (benomyl) and non-systemic (thiram) fungicides to eradicate the seedborne infection to facilitate safe export of seed material, while others for which chemical treatments are not known were detained and destroyed. The entire process of seed health testing and export has been highly successful without even a single omission so far. However, the pathogen diagnostic protocols needs upgrading to include DNA-based technologies and new fungicides tried for eradicating other pathogens.

181 - EFFECTS OF INCUBATION TEMPERATURE AND RELATIVE HUMIDITY ON FUNGAL SPORULATION, MOLD SEVERITY AND SEED QUALITY IN SORGHUM [SORGHUM BICOLOR (L.) MOENCH]

1A 1B 1C 1 1 TONAPI , V.A., RACHNA , M.R., NAVI , S.S., THAKUR , R.P., REDDY , R.K., BANDYOPADHYAY1D, R. & SEETHARAMA1A, N.

1. International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, 502 324, AP, India, [email protected] 1a. National Research Center for Sorghum (NRCS), Rajendranagar, Hyderabad 500 030, AP, India 1b. Department of Botany, University of Pune, 1c. Current address: Plant Pathology Department, College of Agriculture, Iowa State University, Ames, Iowa 50011–1020, USA, 1d. International Institute of Tropical Agriculture, PMB 5320, Ibadan, Nigeria

Grain mold, induced by a number of non-specific fungi, causes substantial loss to seed/grain yield and quality in sorghum [Sorghum bicolor (L.) Moench]. Fungal sporulation and grain mold severity are greatly influenced by temperature and relative humidity (RH) levels. We studied the effects of varying incubation temperatures (25, 27 and 28ºC) and RH levels (85, 90, 95, 96, 97, 98, 99, and 100%) on sporulation of and grain mold severity by three major mold fungi (Curvularia lunata, Fusarium moniliforme, and Bipolaris australienssis) on selected four resistant, four moderately resistant and four susceptible sorghum genotypes. Results indicated both fungal sporulation and grain mold severity increased on most sorghum genotypes with increasing incubation temperature from 25 to 28ºC and increasing RH levels, particularly from 95 to 98%. A linear relationship was obtained between RH levels and grain mold severity and fungal sporulation. The highest sporulation of all the three fungi occurred at 28ºC and 98% RH after 5 days of incubation. Among the three fungi, C. lunata grew and sporulated faster than B. australienssis and F. moniliforme, in that order. Among the sorghum genotypes, IS 25017 supported the least sporulation and had the lowest mold severity, followed by IS 8545 and PVK 801. Seed quality parameters, such as seed germination, seedling vigor index, field emergence potential, Dehydrogenase and λ-amylase activities declined significantly with increasing temperature and RH levels that supported heavy sporulation and grain colonization.

182 - SEED-BORNE FUNGI OF FLOWER PLANTS AND THEIR PATHOGENICITY WU, WEN-SHI, LI, YUEH-LING & WU, HUI-CHEN

Taiwan Hospitality and Tourism College, Hualien County, Taiwan 974

Seeds of 19 different species in 16 genera of flowering crops were health-tested by blotter method in this study. Thirty- four different species of fungi in 20 genera were found among these seeds. Among them, Curvularia brachyspora on sunflower, Alternaria cosmosa on yellow cosmos and Alternaria patula on French marigold were new-recorded seed-borne pathogens. Except ageratum, baby’s breath, salvia, statice, stokesia and pansy, new- recorded seed-borne fungi existed among seeds of the rest species of tested flowering crops.

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183 - DRECHSLERA VERTICILLATA IN BROMUS PICTUS SEEDS, A PATAGONIAN NATIVE POACEA: PATHOGENICITY AND ITS TRANSMISSION FROM THE INFECTED PLANT DEBRIS ZAPATA, RAUL, GAETAN, SILVIA, MADIA, MARTA & MANGIONE, JOSE

Catedra de Fitopatologia.Facultad de Agronomia Universidad de Buenos Aires, Avenida San Martin 4453 (1417). CAPITA FEDERAL, Moldes 1572 . 2 "12" .(1426) Capital Federal, Nuñez 2091. (1429) Capital Federal, AR-ARGENTINA, [email protected]

Bromus pictus (Hook) is a native Poacea that plays a key role in sheep husbandry in the argentine Patagonia. Its natural resowing is affected by a fungus which colonizes the seeds and prevents germination. From naturally infected B. pictus seeds -bearing stromatic structures with conidia- collected in Rio Mayo (45º 25º S, 70º 20º W), Chubut province, Argentina was identified Drechslera verticillata (O’ Hara) Shoem. Its pathogenicity in B. pictus seeds and its transmission from the infected plant debris were verified through several tests. The fungus, cultured in V8 juice and carrot agar plates, developed black olivaceus colonies with stromatic structures. Brown, straight with rounded extremes, 3-7 septated and clear apical cells conidia were observed. They ranged from 60.5µ -16.03 µ to 83 µ -14.8 µ. B. pictus seeds -immersed in sterile water with fungal conidia (104 conidia/ml) and placed in trays with sterilized sand and between filter papers in slides carriers. The experiments were carried out in bioclimatic chamber at 24ºC and artificial light. Inoculated seeds did not germinate while 80% of the controls did. Black stromata with conidia and single conidia on conidiophores were recovered from inoculated seeds. Seeds located close to plant debris bearing fungal stromata had 70% of failures in germination.This is the first report of D. verticillata as pathogen of B. pictus seeds in Argentina and the most austral occurrence of this fungal microorganism.

118 Budapest, Hungary, May 17th – 19th, 2004

6. Seed Improvement Topics: Dormancy breaking; seed sorting; priming; chemical and biological controls; pre- storage treatments; pelleting

184 - THE PROSPECTS FOR SMOKE AND OTHER FIRE-SIMULATION TREATMENTS FOR THE GERMINATION TESTING OF SEED CONSERVATION COLLECTIONS ALI, NATASHA, S. & PROBERT, ROBIN J.

Seed Conservation Department, Royal Botanic Gardens, Kew, Wakehurst Place, Ardingly, West Sussex, RH17 6TN, UK. [email protected]

Seed germination in numerous species has been shown to be stimulated by smoke and its role in the natural regeneration process of plants from fire-prone habitats is unquestioned. However, despite the fact that smoke, applied as an aerosol or as an aqueous solution, is now widely used in habitat restoration projects for stimulating the germination of buried seeds, neither smoke on its own or in combination with other fire-related treatments are widely used in routine seed germination testing in seed banks. Thus a variety of smoke products applied factorially with surgical treatment, potassium nitrate, dry after-ripening and dry heat were tested on seed conservation collections with known dormancy problems. Forty three species from 24 families were tested. Only five collections responded positively to smoke treatment alone, and a further 15 responded to smoke when additional dormancy breaking factors were also present. Twenty one collections did not respond to smoke and in four collections germination was reduced by smoke. The study has shown that smoke treatment could be used as a germination cue in routine viability tests for some species especially when combined with other dormancy breaking factors.

185 - MATRICONDITIONING INTEGRATED WITH CHEMICAL AND BIOLOGICAL SEED TREATMENTS TO IMPROVE STAND ESTABLISHMENT AND YIELD OF CORN ANDREOLI, C. & DE ANDRADE, R.V.

Embrapa Soja, Cx. Postal 231, Londrina, PR 860061-970, Brazil, [email protected]

One of the main reasons of low yield of corn (Zea mays L.) is seed quality, which affects initial stand and plant density. The main objective of this work was to study the integration of matriconditioning with chemical (Captan) and biological seed treatments (Bacillus) on stand establishment and yield of corn, under tropical conditions of Sete Lagoas, MG and under cool conditions of Ponta Grossa, PR., as well as to see if the biopriming could improve the vigour of seed lots with low quality. In order to reach such objectives, two trials were carried out at Embrapa Milho and Sorgo, Sete Lagoas, MG, in 1996 and 1998 and a third one was conducted at Embrapa SNT in Ponta Grossa, PR, in 1997. Seeds of hybrid corn were -1 conditioned in the presence of Bacillus (600 mg.kg of seed), Bacillus+Captan and GA4+7 (100 µM). In the third trial, two seed lots with initial germination of 95% and 85% were conditioned with MC and with MC+Bacillus at rate of 20: 5: 20, at 22ºC for 48 hours. Data were recorded for seedling emergence at 10 and 20 days, final stand, emergence rate, ear number.ha-1, ear and grain yield per hectare. The results showed that the seed treatment with Captan and Bacillus improved stand establishment and yield of 23,4 and 15,8%, respectively, in relation to non-treated seeds. Grain yield of corn did not respond to matriconditioning treatment with GA4+7 at Sete Lagoas, MG. Under cold conditions, the treatment with Captan and MC+Bacillus improved stands but not increased yield of corn hybrids. In the third trial, the initial seed quality affected stand establishment and yield of corn and the integration of MC with chemical and biological seed treatments was effective in ameliorating seed quality and yield of corn, especially under stress conditions. Biopriming with Bacillus seed treatment was effective in improving the vigour of seed lots with low quality.

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186 - EFFECT OF SEED TREATMENTS AND CONTAINERS ON STORABILITY OF SUNFLOWER SEEDS ANKAIAH, R., MANOHAR REDDY, N. & MURALIMOHAN REDDY, B.

National Seed Project, ANG Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, India, [email protected]

Seed of sunflower hybrids APSH-11, LDMRSH-1 and LDMRSH-3 and their parents having above 80% germination and initial seed moisture of 6.5% seed moisture were treated with Metalaxyl @ 6g/kg seed and stored in cloth bag, cloth bag + polythene bag (350 guage) and polythene bag (700 guage) along with control (untreated seed) under ambient conditions of Rajendranagar, Hyderabad, India. Irrespective of genotype and seed treatment, seed stored in cloth bag + polythene bag recorded more than certification standard germination (70%) even after 15 months of storage. Treated seeds stored in polythene bag showed more than 70% germination even after 15 months in all genotypes. Untreated seeds stored in polythene bags had 70% germination upto 6 months in MRHA-1, 207-A, MRHA-2, 7-1B, upto 9 months in MRSH-1, 338A, LDMRSH-3, RHA-271; and 12 months in APSH-11. Treated seed stored in cloth bag showed 70% germination upto 12 months in 338A, MRHA-1, LDMRSH-1, APSH-11 and 7-1A. The seed stored in polythene bag and kept in cloth bag after treatment resulted in higher vigour in all genotypes when compared to seeds stored in remaining treatments.

187 - CHEMICAL-INDUCED SEED POTENTIATION AND FIELD PERFORMANCE OF GRASS PEA AND BLACK GRAM CULTIVARS BHATTACHARJEE*, A., PATI, C.K. & MISHRA, V.K.

Plant Physiology and Biochemistry Laboratory, Department of Botany and Forestry, Vidyasagar University, Midnapore – 721 102, West Bengal, India

Field performance of grass pea and black gram with respect to their growth, metabolic status and yield attributes was analysed from the plants, raised from seeds which experienced pretreatment with two growth retardants (chlorocholine chloride, CCC and Na-dikegulac) and an antioxidant (ascorbic acid). Pretreatment of grass pea and black gram seeds with the chemicals for 6+6 hours (in two instalments) followed by accelerated ageing treatment (99.1% RH , 32 ± 2°C) for 30 days slowed down the ageing-induced rapid loss of germination. The chemical-pretreated seed samples showed T50 (time required for 50% seed germination) values whereas control samples failed to show 50% germination even in extended germination period. The levels of chlorophyll, protein, insoluble carbohydrate, DNA and RNA as well as activity of the enzyme catalase in leaves of the plants (30 days old) developed from the chemical pretreated seeds were found to be significantly higher than control samples. Ageing-induced augmentation of soluble carbohydrate level and activity of IAA oxidase enzyme in leaves of the plants was lower in the chemical pretreated samples than control ones. Plant performance, measured in terms of time (days) required to onset of flowering, fruiting and senescence, was better when they were raised from chemical pretreated seeds. Again, the chemical-pretreated plant samples showed higher yield attributes than control ones and these were recorded in terms of total number of pods per plant, total number of seeds per pod as well as 1000 seed weight. The promising effect of the chemicals on storage potentiation of seeds and subsequent enhancement of plant vigour and crop yield is apparent in this investigation.

120 Budapest, Hungary, May 17th – 19th, 2004

188 - EFFECT OF SODIUM-DIKEGULAC, ASCORBIC ACID AND EUCALYPTUS OIL ON VIGOUR AND VIABILITY OF SUNFLOWER SEEDS UNDER STRESS STORAGE ENVIRONMENT BHATTACHARJEE*, A., CHAKRABARTI, D., KANP U.K. & DAS, R.K.

Plant Physiology and Biochemistry Laboratory, Department of Botany & Forestry, Vidyasagar University, Midnapur-721102, West Bengal, India, [email protected]

An experiment was conducted to evaluate the viability and vigour status of chemical treated seeds of a sunflower cultivar stored under stressful storage environment called accelerated ageing treatment. Sunflower (Helianthus annuus L. cv. MRSF-1051) seeds lost viability at a rapid pace under accelerated ageing condition (99.5% RH, 32±2°C temperature). Pretreatment of the seeds with aqueous solutions of sodium-dikegulac (Na-dikegulac; 2,3: 4- 6-di-O-isopropylidine-∝-L-xylo-2 hexalofuranosate) and ascorbic acid for 8 hours as well as vapour treatment of the seeds with Eucalyptus oil for 60 days slowed down accelerated ageing-induced reduction of seed germinability and TTC (2,3,5-triphenyl tetrazolium chloride) stainability. Reduction of insoluble carbohydrate and DNA levels as well as activities of catalase, peroxidase and dehydrogenase in seed kernels during the forced ageing period was ameliorated to a significant extent in the chemical pretreated seed lots of sunflower. On the other hand, ageing-induced progressive increase of protease activity and levels of amino acids and soluble carbohydrates were remarkably arrested in seed lots pretreated with all the chemicals. Considering the changes of all the vigour indicative physiological and biochemical parameters, the treatments were found to be effective for enhancing vigour and viability of seeds of the sunflower cultivar stored under artificially induced adverse storage situation.

189 - HERBAL MANIPULATIVE METHOD FOR STORAGE POTENTIATION OF MUNG BEAN AND SUNFLOWER SEEDS BHATTACHARJEE*, A. CHAKRABARTI, D., DAS, R.K. & KANP, U.K.

Plant Physiology and Biochemistry Laboratory, Department of Botany & Forestry, Vidyasagar University, Midnapur-721102, West Bengal, India, E-mail: [email protected]

Herbal manipulation of potential status of mung bean and sunflower seeds was critically analysed under artificial stress storage condition. Accelerated ageing treatment (99.5% RH, 32 ± 2°C) on the seeds for 60 days resulted in progressive fall of seed germination with the increase of ageing duration. Seed pretreatment with aqueous plant extracts of neem (Azadirachta indica), kalmegh (Andrographis paniculata) and asoke (Saraca asoka) ameliorated such ageing-induced deleterious effect on seed germinability in both the seed species. Ageing-induced enhancement of seed moisture levels was significant particularly at later observation periods (40 and 60 days after ageing). Changes of germinability and moisture contents of the seeds were associated with the proportional fall of protein, insoluble carbohydrate, DNA and RNA levels in seed kernels with the advancement of ageing duration regardless of the control and treated samples. Seed pretreatment with the plant extracts efficiently retarded the ageing-induced reduction of the biochemical parameters. Conversely, ageing-induced enhancement of the activity of IAA-oxidase was arrested by the seed pretreating agents. Promising effect of the selected Indian herbal agents on storage potentiation of mung bean and sunflower seeds is discussed.

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190 - COMPARISON OF PRIMING AND SUBSEQUENT DRYING TECHNIQUES FOR ONION SEED LOTS: EFFECTS ON GERMINATION AND VIGOUR CASEIRO, ROSELI F., BENNETT, MARK A. & MARCOS-FILHO, JULIO

Universidade de Sao Paulo, ESALQ, Dept. of Producao Vegetal/USP, Av. Padua Dias, 11, P.O. Box 09, 13418-900 Piracicaba, SP, BR-Brazil

The effects of osmopriming (aerated PEG-8000 solution), hydropriming and drum priming on percentage and speed of germination were compared on six lots of onion (Allium cepa) seeds. Osmopriming was performed at -0.5 MPa and -1.0 MPa for 24 h and 48 h. In the hydropriming procedure, seeds were moistened between 2, 4 or 6 layers of paper towel for 48 h or 96 h. The optimal amount of water added and treatment period for the drum priming varied among seed lots. Response to priming varied among seed lots and, in general, less vigorous onion seed lots did not respond well to treatments. The drum priming was efficient for seed hydration but the results showed reduced percentage and speed of germination in most of the seed lots. Hydropriming improved seed physiological potential and was selected for the subsequent study of drying procedures after seed conditioning. Experimental drying procedures included: fast drying at 35°C – 40°C and 40% R.H. for 48h; slow drying (20°C and 50-55% R.H.) for 72 h; seed incubation in PEG-8000 solution or in water bath at 40°C for 1, 3 or 5h or by seed exposure to 35°C for 1 or 2 days. After the slow drying and all incubation procedures, seeds were submitted to fast drying until they reached 6.0% to 8.0% water content. Effects of drying on primed seeds were evaluated by germination, accelerated ageing and electrical conductivity tests. Results identified the fast drying method as the most efficient to preserve the beneficial effects of onion seed priming although only the seed incubation in PEG-8000 solution drastically reduced onion seed germination and vigour.

191 - EVALUATION OF GERMINATION CAPACITY AND PRIMING TECHNIQUES IN SESLERIA VARIA, A PLANT FOR RESTORATION OF CALCAREOUS GRASSLANDS CASTIGLIONI, A., COLOMBO A. & TOSCA, A.

Fondazione Minoprio - Centro Regionale per la Tutela della Flora Autoctona, Viale Raimondi, 54 - 22070 Vertemate con Minoprio - Como – Italy, [email protected]

The purpose of this work is to evaluate priming techniques in Sesleria varia, a promising plant for restoration of calcareous grasslands. Beside its geotechnical proprieties, it helps to recreate highly diverse floristic association. Due to its low germinability and seed production, currently the use of Sesleria varia cannot be applied efficiently in seed mixtures for restoration. Viability and germination capacity of seeds produced ex situ by Fondazione Minoprio, were evaluated. Tetrazolium test was set up and it indicated that seed viability was poor. Gibberellic acid A3 was tested at different concentrations but no statistically significant differences were observed at 48 hours priming. Gibberellic acid priming at 0.29 mM was tested versus the water only or the potassium nitrate solution. Gibberellic acid performed the best germination percentages (over 25%) when priming lasted 24 hours. Even water priming showed a decrease in the germinability with longer times. Potassium nitrate showed the worse percentage of germination, half of those of gibberellic acid at all the durations tested. Mean time to germination decreased with longer times for all the treatments. In comparision with 7 and 14 days, seeds stored for 28 days after priming with gibberellic acid decreased slightly their germination rate and prolonged the time to germination.

122 Budapest, Hungary, May 17th – 19th, 2004

192 - EFFECTS OF SEED OSMOPRIMING ON GERMINATION OF PEPPER SEED AT LOW TEMPERATURES. CAVALLARO, V., IERNA, A. & MAUROMICALE, G.

Consiglio Nazionale delle Ricerche (C.N.R.) Istituto per i Siustemi Agricoli e Forestali del Mediterraneo (I.S.A.Fo.M.), Sezione Colture Erbacee Strategiche di Catania, Via Valdisavoia 5, Catania, Italy, [email protected]

Pepper (Capsicum annum L.) is classified as a warm-season plant requiring relatively high temperatures for successful seed germination. Since the particular climatic conditions of the Mediterranean region permit early productions for pepper, in cold greenhouse, plastic tunnels or seedbeds, sowings are frequently anticipate to late winter or early spring. In these periods, temperatures are equal or lower than minimum for the germination, so seeds take longer to germinate and are subjected during emergence to biotic and abiotic stresses. As a consequence, delayed and erratic plantlet emergence and establishment may result. Seed priming has generally been successful in increasing rate, uniformity, and percentage of germination of many species. Pre-treatment of pepper seed in osmotic solution moreover, hastening the germination and emergence, especially at suboptimal temperatures, would be of significant economic value in the production of greenhouse raised plants. For this reason, our Section carried out a research aiming at studying the effects on pepper germination charateristics of different seed priming treatments with organic and inorganic salts (PEG, KNO3 and KNO3 +K2HPO4) under a wide range of temperatures (from 10 to 25 °C). Two varieties were used. At temperature of 10°C, next to the lower threshold of germination temperature for this species, untreated seed (control) does not germinate, whereas treated seed with organic and inorganic salts showed germination percentage ranging from 20 to 99 %. Even at higher temperatures (13 and 16 °C), where treated and untreated seeds showed a good germination percentage, osmoconditioning with organic and inorganic salts drastically reduced mean germination time.

193 - THE EFFECTS OF POLYETHYLENE GLYCOL ON THE MOISTURE CONTENT AND GERMINATION OF SHELLESS RUBBER SEEDS (HEVEA BRASILIENSIS MUELL-ARG) DURING SIXTEEN DAYS’ STORAGE. CHARLOQ & CHAIRUNNISA HARIS, T.

Fac. of Agriculture, Univ. of North Sumatera, Indonesia

The usual method for transportation of rubber seeds from the Rubber Seed Center to farmers’ plantation is in the form of intact rubber seeds, with the shells, which often raise problems in grading their freshness, and due to bad road conditions, cause the seeds to arrive after more than one week at their destiny, and loose much of their viability. Therefore the Rubber Seeds Center suggested to send shelless rubber seeds, in order to be able to show seed freshness, with much reduction in transportation costs. Since shelless seeds are very prone to drying and damage, a study was conducted with the aim to maintain seed viability during 16 days’ storage. The airdried shelless rubber seeds were immersed for 10 minutes in solutions of PEG-6000 at the concentrations of 0% (control), 15%, 30% and 45%, followed by airdrying, and then the seeds were stored in plastic bags for 4, 8, 12 and 16 days. Seed moisture content after storage, seed germination during storage and seed germination after storage were observed. Results showed that PEG solutions were effective in maintaining the seeds moisture content during storage; after 16 days’ storage, PEG solution at the concentration of 45% showed highest germination (81%) and lowest germination during storage (4.28%), which were even much better after more shorter storage periods, i.e. more than 90% germination and less than 1% germination in storage.

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194 - ANTIFUNGAL ACTIVITY OF 3’, 4’, 7 - TRIHYDROXY FLAVONE ISOLATED FROM BAUHINIA SCANDENS BY CONTROLLING THE IRREGULAR ACTIVITIES OF SOME ENZYMES IN FUNGAL INFECTED SEEDS OF PISUM SATIVUM, PHASEOLUS AUREUS PHASEOLUS MUNGO AND CICER AERIETINUM. CHATTERJEE, PADMA & BARDHAN, ARUNEEMA

Department of BOT ANY, Plant Physiology and Biochemistry Laboratory, University of Kalyani, Kalyani, District, Nadia, Pin Code – 741235, West Bengal. India

The aim of this work focuses an approach towards a non-chemical as well as a non- hazardous control of Fugal infection in seeds of Pisum sativium, Phaseolus aureus, Phaseolus mungo and Cicer aerietinum. The 50% aqueous ethanolic extract of Bauhinia scandens was partitioned over Petroleum ether, diethyl ether and chloroform. Antifungal activity was located in the diethyl ether fraction by spore germination and agar cup methods. An antifungal compound purified by preparative thin layer chromatography was isolated from the diethyl ether fraction and was chemically identified as 3’, 4’, 7 - trihydroxy flavone by uv spectral analysis. The compound (m.p. 255°C) showed absorbance in Methanol at ~max nm 235, 250sh, 309 and 343 and exhibited the characteristic shifts on addition of chemical reagents (NaOAc, NaOAc/H3Bo3, AlCh, AICl3/HCI and NaoMe). Seeds of Pisum sativum, Phaseolus aureus Phaseolus mungo and Cicer aerietinum infected with colony of Fusarium sp and Alternaria sp were pretreated with different concentrations of the above compound. The activities of dehydrogenase, catalase, protease and amylase were studied in non-infected, infected and pretreated seeds. Results indicated that non-infected seeds had low activities of dehydrogenase, catalase, but in the infected seeds there was a sharp rise in their activities. Interestingly enough the pretreated seeds showed low enzyme activities like those of the non-infected Olles. Protease and amylase activities were high in noninfected seeds and low in the infected ones. Here also pretreatment of the infected seeds with the isolated flavone raised the activities of protease and amylase like those in the healthy ones.

195 - SOME EXAMPLES OF BENEFITS BROUGHT BY SEED TREATMENT TECHNOLOGIES FOR DIFFERENT SPECIES GERMINATION DENISE, SOPHIE & BAZIN, MICHELLE

Bayer CropScience Cérès Seed Technology - Méréville France.

A part from well-known benefits of seed technologies such as sowability improvement, delivery of seeds protection products in accurate doses, some enhancement of germination can be obtained by priming, pelleting, film-coating or any specific process. • Priming allows a better germination speed and improves growth homogeneity Pelleting components can be well adapted to a specific species by : - the choice of relevant components, - the size of pellets, - the position of active ingredient(s) Film-coating can improve emergence speed by covering the possible natural cracks of seed coat and/or avoiding some contaminations Process of treatment can guarantee a good storage of the treated seeds by the way of adapted drying.

124 Budapest, Hungary, May 17th – 19th, 2004

196 - TREATMENTS FOR BREAKING DORMANCY OF CEREAL SEEDS DUCOURNAU1, S., GARREAU1, P., FOUGEREUX2, J.A., MERIAUX2, B., AUDE3, L., MALLET1, L. PREVEAUX1, A. & LECHAPPE1, J.

1 GEVES-SNES, rue Georges Morel, BP 24, 49 071 Beaucouzé Cedex, France. [email protected] 2 FNAMS, Le Verger, 49 800 Brain sur l’Authion, France. 3 LABOSEM, Le Verger, 49800 Brain sur l’Authion, France.

Prechilling, preheating and gibberellic acid are methods prescribed by I.S.T.A rules for breaking dormancy of cereal seeds. These prescriptions are not very precise especially for the duration of the prechilling period. Then we have tried to find the shorter duration of a prechilling treatment to break efficiently the dormancy of cereal at the temperature of °C. We also compared the efficiency of the three methods for breaking dormancy. For wheat and barley seed lots tested freshly after harvest, a two days prechilling is sufficient to break dormancy. Results have been reproduced two different years with seed lots of various dormancy levels. The three methods for breaking dormancy were compared on freshly harvested seeds of Triticum durum. On Triticum aestivum, only two methods (prechilling and GA3) were tested. Gibberellic acid at 0.05% is as efficient as prechilling at 5°C during 7 days for Triticum aestivum. For Triticum durum, preheating (seven days at 3°C) is as efficient as gibberellic acid. When seeds are contaminated with Microdochium nivale, poorer germination is observed after prechilling. Theses results show that prechilling at °C duration seems to be reduced at 2 days. The three different methods are both efficient for breaking dormancy, but a 7 days prechilling reduces germination of Triticum durum when applied on seeds contaminated with Microdochium nivale.

197 - MORPHOLOGY AND GERMINATION OF BREU-BRANCO A DORMANT NON-ORTHODOX TREE SEED FROM THE AMAZON. FERRAZ1, I.D.K., GROTH2, D. & COSTA3, M.M.

1National Institute for Amazon Research (INPA), Manaus, AM, Brazil, [email protected]; 2FEAGRI/Campinas State University (UNICAMP), Campinas, SP, Brazil, [email protected]; 3INPA

The genus Protium, from the Burseraceae family, is well known for its aromatic resins occurring in all parts of the trees. Its species are getting recently special attention from the Brazilian cosmetic industry as for example P. heptaphyllum. The fruit (1,8 x 1,5 x 1,3 cm) is a dehiscent red drupe. At maturation the white fleshy mesocarp is exposed, attracts birds and monkeys and completely covers (1,5–3,5 mm thick) the stone. Thus the seed is botanically a pyrene, measuring 1,3 x 0,8 x 0,7 cm with a characteristic semicolon shape and an extremely hard endocarp, which contributes to >60% of the seed dry weight. TSW of fresh seeds directly after extraction from the fruits was 390g and MC 46,7%. The two cotyledons that contain the seed reserves are connected with long petioles to the minute embryonic axis and probably both meristems are rudimentary. Germination of fresh seeds was low 9% and a physical resistance of the endocarp was assumed. The emergence occurs at the sharp end of the seed. Clipping off this end, germination was not increased (9%), as damage of the embryo was probably not avoidable. In a further treatment the seeds were dried on the lab bench for 24 hours with the aim to retract the embryo from the protrusion zone before clipping, thus germination could be increased significantly to 32%. Further drying with or without clipping did not result in any germination, indicating non-orthodox seed storage behavior. Seedling morphology was hypogeal and cryptocotylar. Leave arrangement and leave type was variable, and some seedlings had cataphylls. The first leaves were normally opposite, further leaves spirally arranged. Leaves of Burseraceae family are composed but all seedlings of this species had at least 2 to 6 simple leaves.

125 27th ISTA Congress – Seed Symposium

198 - EFFECTS OF HYDRATION AND DEHYDRATION PERIOD LENGTHS ON THE COLEOPTILES AND ROOT GROWTH AND SEEDLING SURVIVAL OF 3 WHEAT CULTIVARS AFTER REHYDRATION GHORBANI, M.H. ZEINALI, E. & SOLTANI, A.

Dept. of Agron., Gorgan Univ. of Agric. Sci., Gorgan , Iran, [email protected]

Wheat (Triticum aestivum L.) seedlings may meet dehydration between seed imbibitions and emergence, particular in dryland farming. This laboratory experiment was conducted to study the effects of hydration and dehydration period lengths on the coleoptiles and root growth, seedling survival, and their ability to resume growth (after rehydration) of 3 wheat cultivars. Golestan, Khazar-1 and Tajan seed cultivars were germinated for 4, 6 and 8 days in without drought stress condition. Then, seedlings were dehydrated for 2, 4 and 6 days and rehydrated. Number of normal seedling, lengths of coleoptiles and root, and dry weights of seed residues were measured after 7 days. Results showed that when seedlings were grown under suitable moisture conditions for 4 days or less (unpublished data) before dehydration, the length of dehydration periods had no effect on the number of normal seedlings after rehydration, but an increase in the time of dehydration resulted to decries the coleoptiles and root length of 3 cultivars. Dehydration reduced normal seedlings and root lengths, significantly, when seedlings were placed under normal conditions before dehydration for 6 days or more. Reduction in coleoptiles lengths as affected by dehydration was significant only for the treatment 8 days growth under suitable moisture conditions before dehydration. Dehydration period length had no effect on dry weights of seed residues in treatment 4 days growth under suitable moisture conditions before dehydration, while when seedlings were grown under suitable conditions for 6 days, increasing dehydration time followed by significant increase in dry weights of seed residues. In 8 days growth under suitable conditions before dehydration, increasing dehydration period length caused significant increase in seed residues only in Golestan cultivar. These results demonstrate that dehydration tolerance reduces as wheat seedlings develop. On the other hand, the length of dehydration can be effective on the ability of regrowth after rehydration. So this ability decreases as the length dehydration period increases.

199 - MORPHOLOGICAL CHARACTERIZATION OF DISPERSAL UNIT AND GERMINATION STAGE DEVELOPMENT OF CORDIA CURASSAVICA (JACQ.) ROEM. & SCHULT. – BORAGINACEAE. 1GROTH, D., AGUIAR2, R.H., CARNEIRO3, L.M.T.A. & FANTINATTI4, J.B.

FEAGRI/Campinas State University (UNICAMP), Box 6011, Postal Code 13083-970, Campinas-SP, Brazil; [email protected]; [email protected]; [email protected]; [email protected]

Cordia curassavica (Jacq.) Roem. & Schult., known as erva-baleeira, is a native specie from Brazil and has commercial and medicinal importance. The leaves are aromatics and people used it as antiphlogistic. The majority of the native and pioneer species need studies about the propagation methods, morphology, germination, longevity, physiology, conservation and seed health. The objective of these work was make drawings and descriptions of the external and internal characteristics, of the dispersal unit of Cordia curassavica, such as shape, type, size, texture and color of the pericarp and the embryo type, shape, size, texture and position in relation to the storage tissue (endosperm). It was also studied the morphology of the post-seminal development stages. Morphological characterization of the dispersal unit is important to help the identification of a botanical specie, to evaluate and separate species that compose soil seed banks, in the manipulation and conservation of the wild fauna, in study of food supply for herbivorous, in succession and regeneration in forestry ecosystems and natural regeneration of degraded areas. The knowledge of the post-seminal development stages and the morphological aspects of seedlings allows the identification of families, genus and species; are also important in the identification of the plants of a certain region; in studies of the ecology of the specie and of forestry inventory in a region; to knowledge the factors that are responsible for the competition intra and inter specific within a vegetal community and to make easy the interpretation of a germination test in a seed testing laboratory.

126 Budapest, Hungary, May 17th – 19th, 2004

200 - MORPHOLOGICAL CHARACTERIZATION OF SIX SEEDS OF ARBOREOUS SPECIES WITH ORNAMENTAL POTENCIAL. GROTH, D.

FEAGRI/Campinas State University (UNICAMP), Box 6011, Postal Code 13083-970, Campinas-SP, Brazil; [email protected]

The morphological characterization of seeds is an important task to help the identification of a botanical specie in a seed testing laboratory, in soil seed bank and in studies of succession and regeneration in forestry ecosystems. There is a deficiency of specialized bibliography which described seeds of tropical arboreous species. Seeds, like most reproductive structures, are relatively stable under varying environmental conditions and their basic internal organization varies only slightly among related species and genera and the differences that can exist may be regarded as significant phylogenetically and can be used in the taxonomical identification. The objective of this work was make drawings and descriptions of the external and internal characteristics, such as shape, type, size, texture and color of the seed coat and the embryo type, shape, size, texture and position in relation to the endosperm. The studied seeds are: Schinus molle L. (Anacardiaceae R.Br.; occurs from Minas Gerais State to Rio Grande do Sul State); Aspidosperma riedlii M.Arg. (Apocinaceae Juss.; occurs from São Paulo State to Minas Gerais State); Patagonula americana L. (Boraginaceae Juss.; occurs from São Paulo State to the Northwest of Rio Grande do Sul State); Jacaratia spinosa (Aubl.) A. DC. (Caricaceae Juss.; occurs from South of Bahia State to the States of Rio Grande do Sul, Minas Gerais and Mato Grosso do Sul); Lafoensia pacari St. Hil. (Lythraceae Lindl.; occurs in the States of Minas Gerais, São Paulo and Mato Grosso do Sul to Santa Catarina State); Coutarea hexandra Schum. (Rubiaceae Juss.; occurs from Amazonas State to São Paulo State).

201 - EFFECTS OF ELECTROSTATIC FIELD TREATMENT ON PINE SEED GERMINATION GUI, ZHI-BIN & QIAO, LI-MIN

Microelectronics Institute, Xidian University, Xi an, 710071, China, [email protected]

Tree seed of pine, Pinus tabulaeformis Carr., as well as other species, such as tree seeds with shallow dormancy are widely used in mountain and desert areas for reforestation and grassland with airplane sowing seeds in northern China, were treated by electrostatic fields to enhance germination, quantities and quality of young seedling to explore theoretical relationships between electrostatic field treatment and changes in seed vigour. We found that treatment results depended on the form of energy used, treatment dosage, process and index of early stage of seeds. Results showed that the optimal dosage was 500 kV/m for 10 min for treatment of dry or wet seeds for improvement of both germination and root development during initial germination and middle and later stages of seedling development.

202 - EFFECT OF STRATIFICATION IN POLYETHYLENE GLYCOL SOLUTIONS ON GERMINATION OF THREE NORTH AMERICAN SHRUB SPECIES. HARRINGTON, JOHN T. & ROSNER, LEE, S.

New Mexico State University Mora Research Center, Mora, NM 87732, [email protected]

Cercocarpus montanus, Amelanchier alnifolia, and Shepherdia canadensis are North American shrub species with seeds exhibiting physiological dormancy overcome by stratification (moist prechill). Germination of these species occurs during stratification, reducing the number of healthy non-dormant seeds available for sowing at the end of treatment. The effectiveness of incubating seeds in polyethylene glycol (PEG 8000, molecular

127 27th ISTA Congress – Seed Symposium weight 8000u) solutions to reduce germination during stratification was examined for each species using a factorial arrangement of PEG concentrations and 3ºC stratification durations. PEG treatments reduced both moisture content and germination during stratification in a dose-dependent manner for each species. Incubation in PEG at concentrations necessary to suppress germination during stratification had differential effects on the total germination of each species, ranging from no reduction in total germination of A. alnifolia to a large reduction in the total germination of S. canadensis. A discussion of reduced oxygen availability in PEG stratification solutions and its effect on metabolic processes leading to dormancy release or secondary dormancy induction is presented. Species variability in response to stratification in PEG is discussed in terms of this effect.

203 - SEED QUALITY EVALUATION OF VARIOUS DENSITY FRACTIONS OF POLYMER COATED COTTONSEED HOPPER1,2, N.W., OLIVIER1, D.B. & WEDEGAERTNER3, T.C.

1Texas Tech University, 2Texas Agricultural Experiment Station, and 3Cotton Incorporated

Currently cottonseed is prepared for planting by the process of delinting (removal of lint and linters). This is accomplished by exposing the “fuzzy” seed to acid. Following hydrolysis and removal of the fibers, the seeds are passed over a gravity table to remove the low density (quality) seed, treated with various chemicals, and bagged for transport. Delinting using acid poses certain hazards to the seed, workers, machinery, and the environment. Studies are currently being conducted where the longer fibers are removed by mechanical delinting (leaving the short linter fibers) followed by coating the seed and linters with a polymer to allow for seed flowability. To be a viable system the polymer coating must not impede the gravity separation (removal) of the low quality seed and must not slow or impede germination and emergence. Four polymer treatments were imposed on mechanically delinted seed of three cotton cultivars. Subsequently the seed were density separated into three fractions (light, medium, and heavy) using an air separation system. Viability and vigor tests performed on the seed indicated that polymer coated seed could be effectively separated such that an inferior low density fraction could be separated from the higher quality medium and heavy fractions. The data further suggest that polymer coating of the seed up to 2% of the seed weight did not reduce the rate or total germination and emergence.

204 - EFFECTS OF PRE-SOWING TREATMENTS ON EMERGENCE CHARACTERISTICS OF TWELVE TRUE POTATO SEED (TPS) GENOTYPES. IERNA 1, A. & TENORIO 2, J.

1Consiglio Nazionale delle Ricerche (C.N.R.) Istituto per i Sistemi Agricoli e Forestali del Mediterraneo (I.S.A.Fo.M.), Sezione Colture Erbacee Strategiche di Catania, Via Valdisavoia 5, Catania, Italy, e-mail: [email protected] 2International Potato Center (C.I.P.), A.P. 1558, Lima 12, Peru

Potato propagation, carried out traditionally by seed-tubers, produces some disadvantages due to difficulty and high trasport costs of the seed-tubers and also to their short storage time and unreliable phitosanitary status. These disadvantages are particularly important in developing countries where they limit production and consumption of potatoes. True potato seed (TPS) is considered an alternative propagation method able to limit or eliminate some disadvantages linked to seed-tuber utilisation. TPS may show, however, poor percentage and uniformity of germination and low seedling vigour due to dormancy. The effects of seed treatments with some substances, such as gibberellic acid and potassium salts have been investigated, but a large variability of effectiveness has resulted in relation to various causes: genotype, stage of seed development, air temperature during emergence, seed age etc. For this reason it seemed useful to study the effects of presowing treatments with gibberellic acid - GA3 (1500 ppm) and KNO3+K3PO4 (–1.0 MPa) solutions on emergence characteristics and seedling growth of twelve TPS genotypes originated by

128 Budapest, Hungary, May 17th – 19th, 2004 potato crosses in two Peruvian localities: La Molina and San Ramon, characterised by different climatic conditions. Both pre-sowing treatments increased percentage, rate and uniformity of emergence and dry weight of seedlings. This positive effect was more evident in La Molina than in San Ramon. KNO3+K3PO4 treatment determined higher velocity and uniformity of emergence, and consequently greater seedling dry weight too, as compared to the GA3 treatment. The results showed the very important role of genotype and female parent in determining the different response to pre-sowing treatment.

205 - EFFECT OF PRE-SOWING TREATMENT (PST) ON SEED GERMINATION, ESTABLISHMENT AND YIELD. JAGADISH, G.V.

Indo American Hybrid Seeds(I)_Pvt.Ltd.,Bangalore,India

Pre-sowing treatment (PST) on seed lots of Tomato (Cv. Pusa Ruby), Capsicum (Hybrid Bharath) and Onion (Cv. Pusa Red) with GA3 200 ppm and 100 ppm (24 hrs), Kinetin 50 ppm (24 hrs); K2HPO4 and Na2HPO4 at 150 mM (48 hrs), KN03 200 mM (48 hrs) and PEG-1.2 MPa (7 days) and Mannitol –1.24 Mpa (6 days) and hydration – dehydration (48 hrs) at 20°C caused for an increased rate of germination, hypocotyl length and emergence in the field compared to untreated seeds. In Tomato, PST with GA3 (200 ppm), KNO3 (200 mM), K2HPO4 & Na2HPO4 (150 mM), Mannitol (–1.24 Mpa) significantly increased establishment, plant growth and number of leaves at all stages of growth. Kinetin (50 ppm) and hydration and dehydration treatment recorded significant increase in plant height and foliage. Further, significant increase in number of fruits and fruit weight per plant and total fruit yield were obtained with these PSTs. In Capsicum Hybrid Bharath, significant increase in establishment (with GA3 200 ppm, KNO3 200 mM and Na2KPO4 150 mM); growth and number of leaves (with GA3 200 ppm, Kinetin 50 ppm, KNO3 200 mM, Mannitol –1.24 Mpa; hydration-dehydration, K2HPO4 and Na2HPO4 at 150 mM,) were found. On the contrary, significant yield increase was recorded in PST with GA3 (200 ppm), KNO3 (200 mM) and Na2KPO4 (150 Mm), PEG -1.2 MPa .In case of Onion Cv. Pusa Red; plant stand, growth and number of bulb leaves were significantly greater using PST with GA3 200 ppm, KNO3 200 mM , K2HPO4 and Na2HPO4 at 150 mM, Mannitol at –1.24 MPa and PEG – 1.2 MPa. Hydration-dehydration did show significant difference in plant height and bulb leaves only. Significant bulb yield was recorded using PSTs GA3 at 200 ppm, Kinetin 50 ppm, KNO3 at 200 mM, K2HPO4 and Na2HPO4 at 150 mM, Mannitol at –1.24 MPa and PEG –1.2 MPa. The increase in fruit yield was possibly due to the initial advantage of higher percent increase in establishment, growth and foliage.

206 - THE INFLUENCE OF AERATED HYDRATION SEED TREATMENT ON TOMATO SEED INVIGORATION KHAJEH-HOSSEINI1, MOHAMMAD & POWELL1, ALISON A.

1Department of Agriculture & Forestry, University of Aberdeen, Hilton Campus, Hilton Place, Aberdeen, AB24 4FA, Scotland, U.K

Single seed lots from two varieties of tomato, Falat C-H and Early Urbana Y were identified as having low and high vigour, based on the initial assessment of laboratory germination (LG) and controlled deterioration (CD) vigour test (20% mc and 45oC for 24 h). Seeds of each lot were also aged at 20% mc and 45oC for 24h. Aerated hydration (AH) treatment of unaged and aged seeds was carried out for 6 to 48 h in perspex columns with deionised water, placed in a growth cabinet at 20oC and aerated by an aquarium pump. Both unaged and aged seeds of Falat (low vigour) seeds had 93% germination with 40% and 16% normal seedlings respectively. Unaged Early Urban Y (high vigour) had 99% germination and 96% normal seedlings, while aged seeds had 95% germination and 90% normal seedlings. AH treatment improved the number of normal seedlings in Falat C-H from 40% to 93% and from 16% to 91% in unaged and aged seeds respectively. In contrast AH treatment had no 129 27th ISTA Congress – Seed Symposium clear effect on Early Urban Y seeds improvement. Therefore, AH treatment clearly improved the vigour of low vigour seeds (cv. Falat C-H) in terms of the number of normal seedlings. This observation could be explained by the repair of the deterioration that had led to the low vigour and high proportion of abnormal seedlings in Falat C-H seeds.

207 - EFFECT OF CHILLING TEMPERATURE, CHILLING DURATION AND INCUBATION TEMPERATURE ON SEED GERMINATION CHARACTERISTICS OF CEDRUS DEODARA [(G.Don) D.Don)] MUGHAL1, A.H. & THAPLIYAL2, R.C.

1. Division of Forestry SKUAST (K), Shalimar, Srinagar, Jammu & Kashmir, India, 190011. [email protected] ; [email protected] 2. Division of Silviculture, FTSL, F R I, Dehradun, Uttranchal, India, 248006.

The present study on effect of chilling and incubation temperature on various seed germination parameters of Cedrus deodara made it clear that the seeds responded to chilling and as a result of dormancy break due to chilling all the parameters viz. germination, germination value and mean germination time excelled. Seed sources revealed significant differences with maximum germination percentage and germination value after pretreatments being recorded at Srinagar. Minimum germination was recorded at Panchalthan in South Kashmir. Mean germination time also recorded the same pattern. From the point of view of chilling temperature 3°C proved to be significantly better compared to chilling at 9°C. However, northern and eastern aspect seed sources responded well to chilling at 9°C also, while as southern and western aspect seed sources responded well to chilling at 3°C only probably due to the fact that under natural conditions if higher chilling temperature will be required then probably the seeds will germinate after rains in winter on warmer slopes, which will not be in the interests of the seedlings as they are liable to get damaged due to frost in winter months. Besides with the increase in chilling duration all the parameters excelled. Incubation temperatures also recorded significant differences on the germination percentage, mean germination time and germination value of seeds. 30°C incubation temperature proved significantly better followed by 30/20°C. All the two-way interactions studied revealed significant differences. Interaction between incubation temperature and chilling temperatures revealed that 30°C incubation temperature and a chilling temperature of 3°C is significantly better for all the recorded parameters. Interaction between chilling duration and chilling temperature recorded better values at 3°C chilling with duration of 4 weeks. Interaction further reveals that if chilling is to be prolonged then either of the two chilling temperatures i.e. 3°C and 9°C are effective in breaking dormancy. Similarly, interaction between incubation temperature and chilling duration revealed that with the increase in length of stratification seed develops a capacity for germination at lower temperature, which explains the reason for the seeds dispersed in autumn to germinate in spring when temperatures are low.

208 - CHARACTERIZATION OF WATER STATUS IN PRIMED SEEDS OF TOMATO (LYCOPERSICON ESCULENTUM MILL.) BY SORPTION PROPERTIES AND NMR RELAXATION TIMES. NAGARAJAN1, SHANTHA, PANDITA2, VINOD K. & MODI2, B.S.

1Nuclear Research Laboratory, [email protected] 2Regional station, Indian Agricultural Research Institute, New Delhi-110 012, India

The enhanced laboratory and field emergence characteristics of osmo and halo primed tomato seeds (cv. Pusa Ruby) were related to changes in hydration-dehydration kinetics, modified sorption properties and NMR relaxation behaviour of humidified seeds. The hydration-dehydration rates were lower in primed seeds thereby suggesting the protection of seed coat membrane integrity. Water sorption isotherms were constructed for primed and unprimed seeds by equilibrating over different water activities (aw) at 25°C. The analysis of the isotherms by D’Arcy-watt equation revealed that priming has reduced the number of

130 Budapest, Hungary, May 17th – 19th, 2004 strong binding sites and the associated water content significantly and increased slightly the number of weak binding sites and the associated water content. This re-distribution of water that increased the molecular mobility and availability of seed water may be the reason for the higher speed of germination of primed seeds. The changes in transverse relaxation time T2 of seed water and its components measured in vivo using nuclear magnetic resonance spectroscopy showed interesting difference between primed and unprimed seeds. With increase in humidification time, the T2 of primed seeds could be resolved into three components with varying mobilities and the control seeds had only two components till 10 days of humidification. The third fraction with very low molecular mobility that appeared only in primed seeds and accounted for about 50% of proton population was assigned to embryonic tissue water. The relaxation times of other two components of seed water identified as cytoplasmic bulk water and extra-cellular free water also showed higher values in primed seeds. Hence we propose that better performance of primed seeds may be attributed to the modifications to seed water binding properties and re-organization of seed water so as to hydrate the embryonic tissue and to increase molecular mobility of cytoplasmic bulk water required for various metabolic activities during germination process.

209 - THE SHORT-TIME STORABILITY OF VEGETABLE SEEDS AFTER HYDRATION TREATMENT PAZDERA, J.

Czech University of Agriculture in Prague, Department of Plant Production, Kamýcká 957, 165 21 Prague, Czech Republic, [email protected])

Storability of vegetable seeds after hydration treatment was evaluated in this experiment. Two seed lots of radish, onion, carrot and spinach were treated by prehydration and by osmotic priming in PEG 6000 solution. The seed lots were dried after treatment and stored in plastic boxes in 20°C and -18°C for 0, 2 and 6 month. After this periods seed germination, MTG and germination energy were tested. The differences between treated samples of all four crops and between their untreated controls after six month were only small and non – significant. The significant differences between seed lots stored in different temperatures after six month were not registered. Generally is possible to say that short storage of these seed lots after hydration treatment had not significant influence on seed parameters after storage.

210 - ENHANCING SEED GERMINATION RATE OF FOUR TURFGRASS GENERA BY ACID TREATMENTS SALEHI, H. & KHOSH-KHUI, M.

Department of Horticultural Science, College of Agriculture, Shiraz University, Shiraz, Iran, [email protected]

Turfgrasses are commonly propagated by seeds. In some genera of turfgrasses, presence of low germination percentages or rates are the main problems in seed propagation. The present investigation was undertaken to determine the best concentration and duration of sulfuric acid treatment for seeds of following turfgrass genera: Cynodon dactylon [L.] Pers.; Festuca rubra L. var. rubra ‘Shadow’; Lolium perenne L. ‘Barball’ and Poa pratensis L. ‘Merion’. In the first experiment, sulfuric acid was used with concentrations of 0 (control), 25, 50, 75 and 100% for 10, 15, 20, 25 and 30 min. In the second experiment, effects of light/dark (16/8) or dark conditions on germination rates of different mentioned genera were investigated using the best specific acid treatments. Each treatment was replicated 4 times with 100 seeds per replicate kept in plastic petri dishes. Results indicated that in Cynodon, using 50% acid for 20-25 min had highest germination percentage (GP) and mean daily germination (MDG) in light/dark; in Festuca, using 25% acid for 15 min had highest GP and MDG in light/dark; in Lolium using 50% acid for 10 min had highest MDG in dark and in Poa, using 50% acid for 15-25 min had highest MDG in dark. Both Cynodon and Poa had the highest and Festuca had the lowest GPs. Cynodon and Festuca had the highest and lowest

131 27th ISTA Congress – Seed Symposium

MDGs, respectively. This simple procedure, with considerable results, may be replaced to time-consuming and expensive methods such as seed priming.

211 - QUICK RELEASE OF GOLDENRAIN-TREE (KOELREUTERIA PANICULATA LAXM.) SEED DORMANCY SHI RHA1, EUI, REHMAN2, SHAFIQ, PARK3, IN-HWAN &. KIM2, JIN-KEY

1Department of Life Resources Science Plant Genetics & Breeding, Sunchon National University, Sunchon, 540-742, Republic of Korea 2Faculty of Bioresources Science, College of Agriculture, Chonbuk National University, Chonju 561-756, Republic of Korea. 3Department of Landscape Architecture, College of Agriculture, Kyungpook National University (KNU), Taegu 702-701, Republic of Korea.

Goldenrain-tree (Koelreuteria paniculata Laxm.) is wide spread as an ornamental tree and is mostly propagated from seeds but like other temperate trees, it has pronounced seed dormancy. K. paniculata seeds exhibit both seed coat and embryo dormancy (Rehman and Park, 2000) which changes with maturity of seeds (Park and Rehman, 1999). It was found that unscarified seeds failed to germinate after any pre-treatments. Similarly, seeds soaked or soaked and redried without chilling were unable to germinate. However, seeds that were scarified, without soaking or soaking-redrying, gave 44% germination after 60 days of moist chilling. The germination was increased to more than 50% when seeds were soaked or soaked-redried in DW or GA for 5 h and moist chilled for 60 days (DW) and 30 days (GA). On the other hand, the germination of seeds soaked for 24 h and moist chilled was very low, but was higher if the seeds were redried after soaking. Dry chilling after soaking or soaking- redrying for 24 h also promoted germination, and maximum germination (>50%) was achieved after 15 days of dry chilling. In general, higher germination was associated with higher germination rate, i.e., faster germination, except dry chilled seeds that were slow to germinate despite their higher germination. These results clearly indicated that K. paniculata seeds soaking and soaking-redrying prior chilling increased the germination and reduced the time of dormancy breaking compared to conventional chilling treatment.

References Park, I.-H. and Rehman, S. (1999). Studies on seed dormancy: Seeds maturation in relation to dormancy in goldenrain-tree (Koelreuteria paniculata Laxm.). Acta Horticulturae (ISHA) 504: 199-208. Rehman, S. and Park, I.-H. (2000). Effect of GA and chilling on the dormancy of goldenrain- tree (Koelreuteria paniculata Laxm.) seeds. Scientia Horticulturae 85: 319-324.

212 - STUDY OF SEED DORMANCY, AFTER-RIPENING, AND POLYPEPTIDE PATTERNS OF DORMANT AND NON-DORMANT SEEDS IN IRANIAN WHEAT (TRITICUM AESTIVUM) TAVAKKOL-AFSHARI, REZA & JETEZADI-JAMAVAD

Department of Agronomy and Plant Breeding, College of Agriculture, University of Tehran, Karaj, Iran. [email protected]

Pre-harvest sprouting seriously reduces durum grain quality and is considered as an important grading criterion in all market classes of wheat (Triticum spp.). In most cases the major factor being utilized in breeding programs is seed dormancy. To study seed dormancy and after-ripening, 30 bread wheat cultivars were grown under field conditions in a randomized complete block design (RCBD) with three replications. This study was conducted at the University of Tehran’s Research Farm (Karaj) in 2001 and 2002. Days to 50% heading, 50% anthesis, and Zadok's growth stage 92 (ZGS92) were recorded for all cultivars. At ZGS92 spikes were harvested and the grain moisture content was measured for all cultivars. Seed dormancy test was conducted at two temperatures (10°C, 20°C) and seed dormancy index was calculated. To determine the length of seed dormancy, after-ripening tests conducted for six weeks. For measuring α-amylase activity, the falling number test was

132 Budapest, Hungary, May 17th – 19th, 2004 used. Results showed that there were significant genetic variations among Iranian bread wheat for all aforementioned traits. These variations can be used in breeding programs for improvement of pre-harvest sprouting resistance in wheat. To test the hypothesis that differential protein synthesis is responsible for maintaining seed dormancy in wheat, we compared the polypeptide patterns of dormant and non-dormant embryos of wheats. The results demonstrate that steady-state levels of specific proteins differ between dormant and non-dormant embryos during early imbibitions, and these differences may be associated with maintaining and/or breaking seed dormancy.

213 - THE ROLE OF OSMOPRIMING ON GERMINATION OF SUGARBEET SEEDS UNDER DROUGHT STRESS TAVAKKOL-AFSHARI, REZA & JALILIAN, ALI

Department of Agronomy and Plant Breeding, College of Agriculture, University of Tehran, Karaj, Iran. [email protected]

Seed priming is a technique to improve seed and seedling performance. The purpose of priming is to reduce germination time and improve stand and percentage of germination. This research was counducted under laboratory conditions at University of Tehran in 2002. In this study sugar beet seeds of two geneticaly monogerm varieties (Rasol and 276) were selected and primed with polyethylene glycol (PEG) over various times. After determining the optimum potential and time for priming, the primed and control (unprimed) seeds tested under simulated drought stress using PEG solutions (0, -2, -4 and –6 bars). Germinated seeds were counted every 24 h for 10 days. Also the number of normal seedlings were detected. The result showed that the best osmotic potential for priming of sugar beet seeds was -1.2 Mpa for 5 days, which increased germination up to 8% and 14% for 276 and Rasol varieties, respectively. Drought stress decreased germination percentage for both primed and non primed seed, however, this reduction was lower for primed seeds compared to non primed seeds. Mean germination time (MTG) was affected by drought stress, but primed seeds reached their 50% germinationin 0.7 to 1.4 days sooner.

214 - STUDIES ON SEED COLORING IN CEREALS, PULSES, OIL SEEDS AND TOMATO TONAPI1, VILAS A., BABU P.2, HARINATH ANSARI2, N.A. & SEETHARAMA1, N.

1National Research Centre for Sorghum, Rajendranagar, Hyderabad, Andhra Pradesh 500 030, India; [email protected] 2Department of Genetics and Plant Breeding & Seed Science and Technology, Acharya N G Ranga Agricultural University, Rajendranagar, Hyderabad, Andhra Pradesh 500 030, India

The investigations on seed colouring were conducted with Paddy, Sorghum , Maize, Bajra , Redgram, Blackgram, Bengalgram , Castor , Sunflower , Safflower , Soybean and Tomato encompassing 25 dyes namely Rhodomine-B, Cotton blue, Fuchsine, Neutral Red, Gentian Violet. Methylene blue. Crystal violet, Congored, Fast green, Bromocresol Purple, Phenol red, Nigrosine, Erichro black T, Ammonium purpureate (mureoxide), Bromocresol green, Malachite green, Methyl red, Methyl orange, Tital yellow, Indigo carmine along with commercially available natural dyes in the market namely Kumkum, Yellow, Pink, Blue and Brick red to develop color standards. The dye solutions were prepared at 0.75% concentration by dissolving 0.25 g of dye in 16.5 ml water and 15.0 ml ethylene glycol. The various dyes had significant promoting as well as inhibitory impact on mean seed germination (95-21%), vigor index (3349 – 271), field emergence potential (94 – 17.6%) and speed of germination (38.0 – 0.33) and their performance under stress tests across crops and cultivars under investigation. Electrical conductivity of seed leachate was in the range of 128 to 431 µmhos/cm and α-amylase (7.0 – 5.16 mm) and Dehydrogenase activity (0.4 to 0.32) were also affected. The seed germination in exhaustion test significant declined (81 – 20.6%) significantly. Rhodamine-B, Neutral red, Crystal violet. Fast green, Fuchsine, Mureoxide, Congro red, Phenol red and Gentian violet are the best suited dyes for seed coloring, since these dyes

133 27th ISTA Congress – Seed Symposium enhanced seed and seedling characters, enzyme activity. The most deleterious dyes were natural dyes Brick red, Pink, Blue and Kumkum. Other dyes were in mid range.

215 - HIGH STORAGE TEMPERATURE EFFECTS ON BRACHIARIA BRIZANTHA SEED DORMANCY RELEASING USBERTI, R.

Plant Protection Agency, Caixa Postal 960, CEP 13.073-001, Campinas, Brazil. [email protected]

The genus Brachiaria presents a double mechanism of seed dormancy, which could be partially overcame either by H2SO4 scarification or after an open-storage period. This mechanism affects germination percentage results, which are most of the times lower than those obtained by the tetrazolium test. Brachiaria brizantha (palisade grass) seeds show a wide application to set pasture fields, due to a good adaptation and a high forage production. The main aim of this research was to check the effect of high storage temperatures on seed dormancy releasing. Four 1996 B. brizantha seed lots were analysed under 8 constant moisture content (varying from 3% to 18%) and 3 temperatures (40, 50 and 65°C). Twelve subsamples for each seed lot / moisture content / temperature were sealed in aluminium foil packets (250 seeds) and stored in incubators maintained at 40, 50 and 65°C (±0.5°C) during variable periods, from nine minutes to 200 days. The germination tests were performed using 4x50 intact and scarified seed replicates. The hazardous effect of those high storage temperatures seemed to be compensated by higher seed dormancy releasing, increasing germination percentage until a specific point, depending of the temperature and storage period; however, from that point on, the effect of high temperature was deleterious to seed germination. The best seed dormancy releasing results were achieved at 50°C, not taking into account seed moisture contents.

216 - EFFECTS OF SOLID MATRIX PRIMING ON FREE AMINO ACID ACCUMULATION AND WATER STATUS IN LOBLOLLY PINE (PINUS TAEDA L.) SEEDS WU1,2, LUGUANG, HALLGREN1, STEPHEN W., MORT4, ANDREW HUANG1, YINGHUA CONWAY3, KENNETH E. & PAN1,2, YUNRONG

1Department of Forestry, Oklahoma State University, Stillwater, OK 74078, USA, [email protected] (2present address: Botany Department, The University of Queensland, St. Lucia, Qld 4072 Australia); 3Department of Entomology and Plant Pathology, Oklahoma State University, Stillwater, OK, 74078, USA; [email protected] 4Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK 74078, USA

Experiments were conducted to approach solid matrix priming (SMP) effects on free amino acid accumulation and on water status in seed tissues of loblolly pine (Pinus taeda L.). Concentrations of most free amino acids increased after two d SMP and remained high till the late stage of SMP in embryo. Proline, abundant in non-primed seed tissues, increased 2.5- fold at four d SMP, which contributed more than 50% of the osmotic adjustment generated by total amino acids. Consistently, water potential and osmotic potential decreased in embryo during SMP. In addition, all free amino acids content increased after four d germination in both embryo and megagametophytes. Pre-priming treatment of seeds with thiol protease inhibitors (E64, CuCl2, or HgCl2) diminished the SMP effects on free amino acid accumulation, osmotic adjustment, and enhancement of thiol protease activity. The results suggest that thiol protease may be induced by SMP and involved in protein degradation that generates free amino acids for osmotic adjustment and seed invigoration.

134 Budapest, Hungary, May 17th – 19th, 2004

217 - SOLID MATRIX PRIMING OF LOBLOLLY PINE (PINUS TAEDA) SEEDS REVERSES AGING EFFECTS AND OVERCOMES SEED COAT RESTRAINT TO GERMINATION

WU1,2, LUGUANG, HALLGREN1, STEPHEN W., YINGHUA HUANG1, FERRIS1, DAVID M. & CONWAY3 KENNETH E.

1Department of Forestry, Oklahoma State University, Stillwater, OK 74078, USA (2present address: Botany Department, The University of Queensland, St. Lucia, Qld 4072 Australia); 3Department of Entomology and Plant Pathology, Oklahoma State University, Stillwater, OK 74078, USA

The sensitivity of radicle emergence of loblolly pine (Pinus taeda L.) seeds to reduced water potential (ψ) was studied to determine the effects of solid matrix priming (SMP), the seed coat restraint, and artificial aging on germination rapidity (hydrotime, θΗ,), synchrony (σψb) and tolerance of low ψ. Final germination data was determined for a range of ψ from -1.8 to 0.0 MPa and analyzed by a germination rate model (Gummerson 1986) using probit analysis that was developed for herbaceous plants. This model provided a good fit for the pine seed data. SMP improved final germination at all levels of ψ and improved all germination parameters. Artificial aging reduced the levels of all germination parameters but the effect was reversed by SMP. The seed coat was a major restraint to radicle emergence that could be partially reduced by SMP. The employed model demonstrated the family with heavier seed coat showed greater seed coat restraint. The seed coat restraint appeared to act as a barrier to radicle emergence that could be overcome by the development of embryo turgor beyond a threshold.

135 27th ISTA Congress – Seed Symposium

7. Physiological Basis of Seed Quality Topics: Seed development; desiccation; storage; molecular, cellular and biochemical aspects of germination

218 λ-AMYLASE IN SEED AND SEEDLINGS OF CORN AT DIFFERENT DETERIORATION LEVELS APARECIDA FESSEL1, SIMONE, SADER2 , RUBENS, SERRA NEGRA VIEIRA3, ELISA & DE RESENDE VON PINHO4, ÉDILA VILELA

1Agronomist, Doctorate Graduate Student, Seed Production and Technology FCAV/UNESP University – Campus of Jaboticabal, São Paulo State, (SP),Brasil . Zip Code: 14884-900. [email protected] 2Full Professor Crop Science Dept. – College of Agrarian Sciences. Campus of Jaboticabal UNESP UNIVERSITY. Zip Code: 14884-900. Jaboticabal, (SP), Brazil. [email protected] 3Agronomist, Doctorate Graduate Student, Lavras Federal University (UFLA), Agriculture Dept., Minas Gerais State (MG), Brazil 4Associated Professor, UFLA University, Lavras, Agriculture Dept., (MG), Brazil. [email protected]

This research was carried out with the objective of verifying the λ-amylase activity in corn seeds and seedlings at different deterioration levels. Were used hybrid corn seeds lot 9569 from the Dow AgroSciences Seed Company with a moisture content of 15% and aged for 24, 48, 72, 96, 120 and, 144 hours. For the enzyme extraction were chosen at random 400 seeds and placed to germinate for 70 hours at 25ºC. After this period half of the sample had the seedling removed and macerated at liquid nitrogen and in the other part related to the seedlings was discarded and the seeds ground in a cold mill at 4ºC. Was verified a low λ- amylase in the seedlings and a higher activity in the seeds. Smaller enzymatic activity was observed in aged seeds for a period of 144 hours, indicating a low activity in deteriorated seeds. According to the obtained results was concluded that to detect the λ amylase activity is suggested to use pre-germinated seeds excluding the seedlings, so can be correlated the loss of viability and the decaying of the enzyme activity.

219 - WATER ACTIVITY AS A BETTER APPROACH THAN MOISTURE CONTENT FOR FOREST SEEDS MOISTURE MANAGEMENT. BALDET1, PATRICK, SCHEVIN2, PATRICK, VALLANCE3, MICHEL & VERGER3. MICHEL

1Cemagref, unité Ressources génétiques et Plants forestiers, Domaine des Barres, F-45290 Nogent sur Vernisson. 2Institut des Sciences de l'Evolution, UMR CNRS 5554, Université Montpellier II, place Eugène Bataillon, F- 34095 Montpellier cedex 05. 3INRA, UE 995, Avenue de la Pomme de Pin - Ardon BP 20619, F- 45166 Olivet cedex

Seed moisture is a key factor influencing seed quality and an continual challenge for forest seed industry. Moisture content (MC) is the most common procedure used to assess moisture of forest seeds; for a given species, the range of optimal water content for storage has been defined both by know-how and conducting tests. However, this approach is not always reliable because it does not take into account the chemical properties of water remaining in dried materials. On the contrary, the concept of activity of water (Aw) quantifies the vapour pressure generated by the moisture present in hygroscopic products like organic substances. It is known as the chemical potential of water. Aw has been widely used for decades by food industry. In this discipline it is very well-known that the reactivity and depreciating ability of hydrolytic reactions, enzymes activity, lipid oxidation or biotic agents growth are dependent on Aw rather than on MC. Consequently, Aw is more consistent to define stability moisture for organic materials preservation. The relationship between Aw and MC is pictured by the sorption/desorption isotherm. This connection is rather stable for standardized organic materials like sugars or flours where it is possible to use equally MC or Aw for moisture assessment. Forest seeds are picked from natural stands or seed orchards where a high genetic diversity is sought after.

136 Budapest, Hungary, May 17th – 19th, 2004

The combination of genetic and environmental factors results in unpredictable seed characteristics. The R&D project we carry out in cooperation with French forest seed companies Vilmorin and ONF aims to: a) the experimental construction of sorption isotherms of different seed lots to demonstrate the consequent variability among different seed lots for a given species, b) calculate the water activity of greater stability on the basis of desorption isotherms and in accordance with bibliographic references, c) demonstrate the effectiveness of water activity assessment versus moisture content evaluation d) finally, to transfer the Aw technique to ONF and Vilmorin for seed moisture control. Aw applied to forest seeds moisture management is now regularly being used by Vilmorin and on the way to be by ONF.

220 - MAPPING QUANTITATIVE TRAIT LOCI CONTROLLING SEED LONGEVITY IN MAIZE. CASARINI, E. FRASCAROLI, E.GAMBERINI, G. & CONTI, S.

Seed longevity is a very crucial trait for trade and for conservation of genetic resources in seed banks. It has been proved that defoliation of mother plant during kernel maturation can improve cold tolerance of germinating seeds. This study was aimed to analyse the effect of defoliation on seed longevity and to detect quantitative trait loci (QTLs) involved. For this purpose, a mapping population of 171 F2:3 families was chosen, generated from two inbred lines (Lo1016 and Lo964), differing for cold tolerance at germination and for reaction to defoliation. Plants were selfed in 1998 and seeds were harvested from defoliated (D) or non- defoliated (ND) individuals. Seeds were evaluated for germination in 1999 and then kept at about 12% of moisture, sealed in plastic boxes at room temperature (17°C-30°C). After five years, germination at 25°C was measured. Leaf removal caused a noticeable increase of seed longevity, with different effects on families. Composite interval mapping was used to identify QTL for reaction to defoliation. From a preliminary analysis, five putative QTL were detected in D and five in ND population, explaining about 21% and 33% of variability, respectively. Two of them were in common, and both parents contributed alleles increasing seed germination. Mapping of QTL will allows the detection, localisation and characterisation of genetic factors contributing to seed vigour and longevity. The presence of genomic regions possibly involved in reaction to defoliation and its relation with seed longevity and tolerance to cold is also discussed.

221 - THE EFFECT OF DIFFERENT POST-HARVEST DRYING METHODS ON SEED QUALITY FROM CAPSULES OF FOXGLOVE (DIGITALIS PURPUREA L.) HARVESTED AT THREE STAGES OF MATURITY. CONEYBEER, JULIA, ADAMS, JOHN & PROBERT, ROBIN J.

Seed Conservation Department, Royal Botanic Gardens, Kew, Wakehurst Place, Ardingly, West Sussex, RH17 6TN, UK. [email protected]

Capsules of foxglove representing three stages of maturity were harvested on a single day. For each maturity stage, capsules were divided into four drying treatments. 1) capsules held for 10 d under ventilated ambient conditions prior to seed extraction and drying. 2) capsules open dried in a seed bank dry room (15°C and 15% RH). 3) capsules dried in a cloth bag in the dry room (standard conditions for seeds from dry dehiscent fruits). 4) capsules dried in a ventilated box in the dry room. Overall, the differences in seed quality related to capsule maturity were comparatively small. As expected highest seed quality was recorded in seed extracted from the most mature capsules that had began to dehisce at the time of harvest. Seeds quality from these capsules was not influenced by subsequent drying method. However, seed quality was affected by drying method in less mature capsules where

137 27th ISTA Congress – Seed Symposium relatively slow drying led to highest seed quality. For the least mature capsules, seeds were longest lived in ageing tests when the capsules were held under ambient conditions for 10 d. For capsules of intermediate maturity, seed viability was highest when seeds were dried in a ventilated box.

222 - PHYSIOLOGICAL AND BIOCHEMICAL CHANGES IN COTTON SEEDS DURING DETERIORATION. DIAS, D.C.F.S., FREITAS, R.A., DIAS, L.A.S. & OLIVEIRA, M.G.A.

Department of Fitotecnia / BIOAGRO, Universidade Federal de Viçosa, 36571-000, Viçosa -MG, Brazil, [email protected]

This work was carried out aiming to evaluate the physiological and biochemical changes in cotton seeds during natural and artificial aging. Seeds from two varieties (Fabrika and IAC- 20 RR) were stored during 12 months under environmental condition and in a cool room (13°C). In order to evaluate alterations during artificial aging, the seeds were aged at 42°C and 100% RH for 0, 24, 48, 72, 96 and 120 h. Physiological quality (standard germination, accelerated aging and cool germination tests) and biochemical assays (lypoxigenases, acid phosphatase and tripsin inhibitor activities, and lipids content) were performed at 0, 2, 4, 6, 8, 10 and 12 months storage and after each artificial aging period. The seeds stored under environmental conditions and in cool room were also evaluated by electrical conductivity test and seedling emergence in sand. Deteriorated seeds, naturally and artificially aged, had decreasing on germination, vigor and lypoxigenase, acid phosphatase, tripisin inhibitor activities and lipids content. However, it was not possible to determine correspondence between physiological and biochemical changes in both naturally and artificially aged seeds.

223 - CONSERVATION OF SEEDS FROM PATA-DE-VACA (BAUHINIA FORFICATA). DODE1, LUCIANA BICCA, DA SILVEIRA LAZZAROTTO2, SIMONE ROSA, GONÇALVES3, CIANE XAVIER & ANDRÉ TILLMANN4, MARIA ÂNGELA

1Catholic University of Pelotas, 2BIC/UCPel, 3ProBIC/FAPERGS, 4UFPel, [email protected]

The maintenance of genetic diversity is a determining factor in the propagation of medicinal plants and recovery of [degraded] deforested areas. The arboreal species Bauhinia forficate is frequently used in urban planting to recover these deforested areas; it also has a medicinal value and can be propagated in an efficient form using seeds. Cryopreservation is an alternative method for the long-term conservation of seeds; it drastically reduces the physiological activity of the conserved material at ultralow temperatures, usually liquid nitrogen (-196°C). The aim of this research was to establish a cryopreservation protocol and determine the effect of cryoprotectants during storage. The seeds were submitted to 6 treatments: negative control; cryopreservation in liquid nitrogen; liquid nitrogen and the cryoprotectant glycerol (50%); glycerol (50%) only; liquid nitrogen and osmotic pre- conditioning (glucose 0.4 M); osmotic pre-conditioning (glucose 0.4 M) only. The process of cryopreservation was carried out for 1.5 h in liquid nitrogen and the samples defrosted in a waterbath at 37°C for 10 minutes. The seeds were then tested for the ability to germinate at 28°C in damp paper rollers; the seeds were evaluated at 7, 14, 21 and 28 days. 28 days after planting the highest percentage of germination was observed in those seeds osmotically pre-conditioned and those stored in liquid nitrogen (91% and 89% respectively). The lowest percentage of germination was observed in those samples osmotically pre-conditioned and cryopreserved in liquid nitrogen (73%). The seeds from Pata-de-Vaca can be stored at ultralow temperatures without changing their ability to germinate.

138 Budapest, Hungary, May 17th – 19th, 2004

224 - THE INVESTIGATION OF MAIZE SEEDLINGS GROWTH ORIGINATED FROM AGED OF SEEDS. I RELATIONS OF MASS AND PHOSPHORUS

DRAGIČEVIĆ1, V., SREDOJEVIĆ1, S., VRVIĆ2, M., IVANOVIĆ3, M., ĐUKANOVIĆ1, L. & KOLAR- ANIĆ4, L.J.

1Maize Research Institute "Zemun Polje", Slobodana Bajića 1, 11185 Zemun Polje, Serbia and Montenegro 2Faculty of Chemistry, University of Belgrade, 11001 Belgrade, Studentski trg 16, P.O.Box 158, Serbia and Montenegro 3Institute for, Maksima Gorkog 30, 21000 Novi Sad, Serbia and Montenegro 4Faculty of Physical Chemistry, University of Belgrade, 11001 Belgrade, Studentski trg 16, P.O.Box 158, Serbia and Montenegro

The investigation was set up to study mass and phosphorus (carrier of bounded energy) partitioning in seedlings influenced by accelerated ageing of maize hybrid seeds (dent and sweet maize). The damages originated from seed ageing influenced the drop of germination percentage with reduced growth of root and shoot via mass and phosphorus retaining in seed rest of seedlings. As well, root and shoot had lower concentration in relation to seed rest, with lesser hydrolysis and biosynthesis. The behaviour of seedlings was similar to conditions of low phosphorus nutrition. They could growth by different mass and phosphorus distribution in relation to seedlings originated from non-aged seeds – in direction to root domination. In the hybrid with better mass and phosphorus efficiency (dent maize) growth realizes with phosphorus allocation to root and cell volume increasing, opposite to the hybrid with slighter mass and phosphorus efficiency (sweet maize) realizes with dry mass and phosphorus allocation to root. The higher ability to conserve germination with elevated mass and phosphorus efficiency, as it was found in dent hybrid could be coupled with mechanisms of rational energy utilization in conditions of reduced mass and phosphorus ability. Authors' aim: is exploration of seed ageing and its influence on biochemical reactions in seeds and seedlings originated from aged seeds, including degradation of stored and antioxidative biomolecules. On the other hand lays investigation of stimulative substances on germination and growth processes. Major findings: represents simple methods for screening of seed materials' condition and on this basis classification of genotypes in to groups with “good” and “bad” ability to conserve germination. Then again, there was determined several groups of natural and synthetic substances with hormetic effectiveness on germination and growth process in plants.

225 - THE INVESTIGATION OF MAIZE SEEDLINGS GROWTH ORIGINATED FROM AGED OF SEEDS. II RELATIONS OF WATER AND PHOSPHORUS DRAGIČEVIĆ1, V., SREDOJEVIĆ1, S., VRVIĆ2, M., TODOROVIĆ2, M., ĐUKANOVIĆ1, L. & KOLAR-ANIĆ3, L.J.

1Maize Research Institute "Zemun Polje", Slobodana Bajića 1, 11185 Zemun Polje, Serbia and Montenegro 2Faculty of Chemistry, University of Belgrade, 11001 Belgrade, Studentski trg 16, P.O.Box 158, Serbia and Montenegro 3Faculty of Physical Chemistry, University of Belgrade, 11001 Belgrade, Studentski trg 16, P.O.Box 158, Serbia and Montenegro

The quantification of growth processes from energy point of view in seedlings represents the balance of free energy - water accumulation and bounded energy - utilization of reserve phosphorus from seed. The search was set up to study water and phosphorus partitioning in seedlings influenced by accelerated ageing of maize hybrid seeds (dent and sweet maize). The root and shoot of seedlings originated from aged seeds has elevated water and phosphorus amounts per 1 g of dry mass. It was observed reduced water and enlarged phosphorus share in root but amplified water and declined phosphorus share in shoot. As the initial growth of seedlings dictates increased request for energy amounts, it could be supposed that growth of seedlings originated from aged seeds was realized by support of free energy in shoot and bounded energy utilization in root. The noticed changes were more

139 27th ISTA Congress – Seed Symposium intensive in seedlings of sweet maize hybrid. In thus, the equilibrium between free and bounded energy could represent the key of rational energy use. Authors' aim: is exploration of seed ageing and its influence on biochemical reactions in seeds and seedlings originated from aged seeds, including degradation of stored and antioxidative biomolecules. On the other hand lays investigation of stimulative substances on germination and growth processes. Major findings: represents simple methods for screening of seed materials' condition and on this basis classification of genotypes in to groups with “good” and “bad” ability to conserve germination. Then again, there was determined several groups of natural and synthetic substances with hormetic effectiveness on germination and growth process in plants.

226 - REACTION TO DEFOLIATION AT DIFFERENT TIMES DURING SEED DEVELOPMENT AND TOLERANCE TO COLD AT GERMINATION IN MAIZE. FRASCAROLI, E., CASARINI, E., FELTRAMI, E. & CONTI, S.

A preliminary study made on a few maize inbred lines suggested the presence of genetic variability for the acquisition of tolerance to cold, at germination, of seeds produced by plants subjected to defoliation during seed development. Objective of this study was to ascertain if the genotypic response to defoliation is influenced by the developmental stage in which the treatment is applied. Inbred lines B73, Lo1016 and Lo964 were considered, based on the previous results. One-hundred-and-fifty plants for each inbred were grown in the field, selfed and defoliated at 15, 18, 21, 24, 27, 30, 33, 36 and 39 days after pollination, or non-defoliated (ND). Six hours after defoliation, from D and ND plants, immature seeds were collected for fresh and dry weight measures, and for ABA content analysis. All the traits utilized to describe seed developmental stage pointed out differences among genotypes. In Lo1016, low levels of ABA were detected at all stages, while Lo964 and B73 showed an increase of the hormone content during development. Mature seeds were evaluated for germination at 25°C (G25) and at 9°C (G9). Inbred line Lo1016 showed an improvement of tolerance to cold after defoliation at all times of treatment, both as mean time to germination (MTG at 9°C) and as germinability (G9/G25). The genotypes here investigated could thus represent a suitable material for the genetic dissection of tolerance to cold at germination and for the analysis of the expression of genes involved in the acquisition of the trait.

227 - WATER UPTAKE PATTERN IN VIGNA RADIATA SEEDS DURING INCUBATION UNDER CHILLING TEMPERATURE AND CYCLOHEXIMIDE TREATMENT KAR, RUP KUMAR & CHAKRABORTY, RAJKUMAR

Plant Physiology and Biochemistry Laboratory, Department of Botany, Visva-Bharati University, Santiniketan 731 235, West Bengal, INDIA

Present investigation aimed to study the water uptake by whole seeds (intact or scarified) or by embryonic axes (attached or detached) of mung bean (Vigna radiata) during early incubation and influence of chilling temperature (4ºC) and cycloheximide (10-4 M; CHI) treatment. In case of intact seeds, water uptake by whole seeds started after a lag period of 4 h during incubation at room temperature (30ºC), while no uptake occurred at all at 4º C during early incubation. Scarified seeds showed a faster uptake of water without a lag period at 30ºC and chilling treatment slowed down such uptake. On the other hand, CHI treatment could prevent water uptake by whole seeds very little in case of intact seeds although somewhat prevented in scarified seeds. When intact seeds were preincubated in water at 30ºC for 6 h before embryonic axes (either attached with cotyledons or detached) were monitored for water uptake, incubation at 4ºC totally prevented water uptake by axes. Similarly, water uptake by embryonic axes was prevented also by CHI treatment, although there was an initial rise in fresh weight in case of axes treated with CHI in detached condition. In case of scarified seeds water uptake by embryonic axes (attached and detached) was monitored during incubation at 4º C following a preinubation at 30ºC for 1 h. In control sets water uptake by attached axes was somewhat faster than detached ones.

140 Budapest, Hungary, May 17th – 19th, 2004

Chilling treatment almost prevented water uptake in both cases. Similarly, CHI treatment in case of scarified seeds also prevented water uptake in case of both attached and detached embryonic axes.

228 - RELATIONSHIP BETWEEN ABSCISIC ACID, GIBBERELLINS AND PRECOCIOUS GERMINATION OF MALTING BARLEY KERNELS KLEMŠ, M., HRSTKOVÁ, P., FIŠEROVÁ, H., PROKEŠOVÁ, Z., VLAŠÁNKOVÁ, E. & PROCHÁZKA, S.

Department of Botany and Plant Physiology,Faculty of Agronomy, Mendel University of Agriculture and Forestry Brno, Zemědělská 1, Brno 613 00, Czech Republic

We studied the relationship between preharvest sprouting of kernels in intact ears of barley (cvs. Akcent and Krona) and the activities of the abscisic acid and gibberellins in the stages of wax and full maturity of kernels. Barley ears were incubated for 5 days in a darkened room under 98% relative humidity, after 8-hour treatment with one of the following solutions: 10 µM of abscisic acid (ABA), 20 µM of fluridone, 10 µM of gibberellic acid (GA3) and 30 µM monensin. In the wax stage the kernels of control variants of both cultivars did not grow through the ears. After treatment with gibberellin and fluridone the kernels of the Akcent cultivar grew through (30%). Monensin applications considerably suppressed growing through in ears of both cultivars. In the stage of full maturity growing through was observed in all variants of both cultivars with the exception of those treated with monensin (less than 5%). The dry matter in germinating kernels from ears incubated on monensin in full maturity was higher than from the other variants. Abscisic acid treatment inhibited germination. The level of endogenous ABA decreased in all variants within 5 days of ear incubation in the germinators. Fluridone application did not reduce the ABA content until full maturity compared to the untreated controls. A higher β-glucosidase activity was detected after gibberellin application in the stage of wax maturity. Gibberellin activity was higher after ear incubation on gibberellin, fluridone or monensin in the stage of full maturity of kernels. Acknowledgement: This research was supported by the project of the Czech Ministry of Education (No. MSM 4321 00001).

229 - MICROHARDNESS OF COTTON-SEED HULLS KRAKHMALEV, V.A. & PAIZIEV, A.A.

Institute of an electronics of an Academy of Sciences of Republic of Uzbekistan, Tashkent

Microhardness method is testing tool for difference materials under indentation by a diamond pyramid at small loading (from 1 up to 20 gauss). The impressions, obtained at that, have the microscopical sizes and do not damage the investigated sample. Microhardness of a material is determined by value of a micro volume of an impression on the sample. The maximum efficiency of the microhardness method is reached at the simultaneous microscopical control of an investigated material. In this case it is possible to investigate mechanical properties of separate structural elements, to study and to control physical condition of thin surface layers on the articles, films under different mechanical, chemical and heat treatments. It is rather perspective applying of the microhardness method to agricultural objects. The available information in this respect are rather limited. In particular, in the given work the microhardness method in first is applied to research and testing of mechanical properties of cotton-seed hulls. Till now problems of strength and resiliency of seed cover for difference cotton sorts are studied unsufficiently, and the data about physical-mechanical properties of separate structural layers of seed cover absent in general. On the assumption about an isotropy of properties of a seed cover is theoretically showed, that maximal permissible load, which one can stand the seed without damage of a germ, depends on microhardness of seed cover and main radius of curvatures of a surface in a loading point. According to outcomes of outside epidermis microhardness measurement is computed, that the maximal load for lateral part of the seed surface is 26, for halasal part is 52 and for micropilar part 131 gauss. 141 27th ISTA Congress – Seed Symposium

The analysis has shown, that generally microhardness of cotton-seed hull both in accross - track direction on cotton cover, and along each layer is distributed nonuniformly. The research of an outside epidermal layer, pigmental and columnar layer for microhardness has shown, that 40-45% of cotton seeds have heightened strength of micropilar part, 35-40% have heightened strength for halasal parth and 3-10% of seeds have identical microhardness on the all part of seed cover. And for 10-25% seeds the microhardness of a lateral part is more than microhardness of micropilar and halasal parts simultaneously. It is showed, that in the deep of seed cover there is thin interlayer of a material with anomalously large value of microhardness. Sudden change of microhardness is watched in a thin layer of seed cover, passing through top ends of columnar cells and almost coincident with a so- called light line. The comparison of microhardness for different layers of seed cover with microhardness of some metals demonstrates, that for most mature cotton seeds of a sort 108-Ф an outside epidermis outside pigmental and crystalliferous layers have same hardness, as the annealed aluminum, and columnar layer at a level of a light line have hardness as for annealed brass. As the microhardness now becomes the relevant characteristic of physical-mechanical properties of seed cover and determines number of defects in a fiber at ginning of cotton raw, it is interesting to learn (at identical damp) microhardness of cotton-seed hull of the diversified cotton sorts. Such analysis is made for 22 cultivated sorts and more 10 wild cotton sorts. Such analysis the microhardness of an outside epidermis, selection feature to generate different morphological defects in a fiber, easiness for interruption of microscopical piece of seed cover together with a fiber allows beforehand to forecast clogging up a fiber by seed cover. The analysis demonstrates, that the greatest hardness of an outside epidermis have the sorts 108-Ф, 149-Ф, Аз-512, 9155. the seeds of the majority of the wild cotton have higher microhardness, than cultural. However, for some from wild cotton the hardness of outside epidermis is not heigh . The variety G. harknessii Brandg, G. Trilobum Skovsted and G. Stocksii M. Masi are as example to such cotton sorts. It is remarkable, that the microhardness of a columnar layer is practically constant for seed cover of any sorts and kinds of cotton and has values from 20 up to 22 kg / мм2. From 32 investigated cotton seeds the sort 108-Ф is particular, as its microhardness at the mentioned above level of section is on 3-5 kg / мм2 higher. This fact indicates, that the hardness of cotton-seed hull depends not on its general thickness, and from a ratio of separate thicknesses of columnar layer and its anomalously solid part. The heightened hardness of an abnormal part located near light line, is conditioned by a accumulation degree in a columnar layer of different substances, in particular cutin.

230 - EFFECT OF TEMPERATURE ON THE GERMINATION OF GRAIN AMARANTH LOONAT 1, T.A. VAN DEN HEEVER 2 E. & HAMMES 3,, P.

1National Department of Agriculture, Private Bag X 3917, North End, Port Elizabeth, 6056 South Africa, 2ARC-Roodeplaat, Private Bag X 293, Pretoria, 0001 South Africa and 3Department of Plant Production and Soil Science, University of Pretoria, 0002 South Africa

Amaranthus is produced as a grain or leafy vegetable in many parts of southern Africa and has potential commercial importance. It is commonly known as Thepe (Tswana; Sotho), Imbuya (Xhosa) Utyutu (Zulu) and Vowa (Venda). Several Amaranthus species utilize the C4 photosynthetic pathway and consequently exploit carbon dioxide more efficiently resulting in this crop being more heat and drought tolerant than most agronomic crops. These qualities have created a renewed interest in Amaranthus as an alternative crop. To make optimal use of the growing season, planting under cool soil conditions is often necessary. The objective of this investigation was to establish the effect of temperature (12, 17, 22 and 28°C) on seed germination of A. hypochondriacus (unknown landrace) and A. cruentus (cv. Amar and Anna). Whereas overall germination was advanced by increasing the temperature, only A. hypochondriacus germinated abundantly at temperatures as low as 12°C. A. hypochondriacus had a germination percentage of 70% at 12°C while A. cruentus had a germination percentage of less than 1% at the same temperature.

142 Budapest, Hungary, May 17th – 19th, 2004

231 - STUDIES ON LOCALISATION OF SEED DETERIOATION AND SEED TREATMENTS TO CONTROL SEED DEGENERATION IN SUNFLOWER (HELIANTHUS ANNUS L.) NAGI REDDY, Y., KESHAVULU, K., MANOHAR REDDY, N., ANKAIAH R. & MURALIMOHAN REDDY, B.

National Seed Project, Acharya N.G. Ranga Agricultural University, Rajendranagar, Hyderabad-500 030, India

Seed deterioration causes loss of viability in storage and consequently affects the potential performance under field conditions. The process of seed deterioration starts from physiological maturity onwards and this degenerative process continues until it is planted. Various factors (climate, moisture stress at critical stages, nutrient stress, pest and diseases) affect the initial quality of seed thereby gradually affecting storage potentiality. Four months aged seed lots of APSH 11, sunflower hybrid and its parental lines (7-1A, 7-1B and RHA 271) were treated with thiram @ 2.5 g/kg and invigorated with Na2HPO4 @ 10-3 M and KNO3 @ 0.5% for localisation of seed deterioration studies. Seed deterioration began from the tip of the radicle and extended to hypocotyl and finally to embryonic axis. The most sensitive site prone to seed deterioration in sunflower was radicle, then plumule and cotyledons. The cotyledons were resistant to deterioration. Hydration-dehydration treatments particularly KNO3 @ 0.5% was found effective to prevent release of free radicles, lipid peroxidation and improving the shelf life of the seed.

232 - STORAGE OF SEEDS FROM ALBIZIA LEBBECK (L.) BENTH. UNDER AMBIENT CONDITIONS IN CUBA. NAVARRO*, MARLEN & MESA, A.R.

Seed Production and Technology Research Project. Experimental Station on Grass and Forages “Indio Hatuey”. Matanzas. CP 44280. Cuba. (*[email protected])

With the objective of evaluating the effect of the storage of seeds from Albizia lebbeck (L.) Benth. under ambient conditions in Cuba on viability and germination and thus identifying and characterizing the dormancy period in this species, the performance of these seeds during 11 months in a storehouse was studied, for which a simple classification design was used with four replications of 100 seeds each; and the evaluations were carried out at 0, 2, 5, 7, 9 and 11 months of storage (m.o.s.). The variables measured were: percentage of germination and viability, as well as hard and dormant seeds; variance analyses were performed for comparing the means. It was determined that during the time from harvest to the beginning of storage A. lebbeck seeds are not deteriorated. Or at least the damage is not perceptible; while as physiological age increases, the seeds age progressively but naturally; the decrease of viability followed a sigmoid pattern and an inversely proportional relation was observed between this parameter and the physiological age of seeds; from the 2 m.o.s. there was and induction toward secondary dormancy stages. Key words: Albizia lebbeck, storage, viability, germination, dormancy

143 27th ISTA Congress – Seed Symposium

233 - SEED RESPIRATION IN TWO MACROTHERMAL SPECIES DURING GERMINATION AS AFFECTED BY TEMPERATURE AND OSMOCONDITIONING PATANE’, CRISTINA, D’AGOSTA, GIUSEPPINA, AVOLA, GIOVANNI & IACOPONI ELISA

CNR-Istituto per i Sistemi Agricoli e Forestali del Mediterraneo (ISAFoM) - Sezione Colture Erbacee Strategiche, via Valdisavoia 5, 95123 Catania (Italy) Tel.: +39095234480; Fax: +39095234449. [email protected]

With the aim at studying, in two macrothermal species (sunflower and grain sorghum), the effect of temperature and osmoconditioning upon seed respiration, as an index of its metabolic activity, an experiment was carried out where, starting from initial seed imbibition up to visible radicle appearance, the CO2 released from seeds was monitored. Two temperatures, with 25°C as optimal and 15°C as suboptimal, and two seed treatments, with the first represented by an untreated control and the second one by seeds primed in a 250 gl-1 of Polyethylene glycol 6000 (PEG) osmotic solution, were compared. In both species, osmopriming make faster but not higher the seed germination; at 15°C, the maximum respiration value reached in the two seed treatments was similar but in primed seed it was recorded earlier than control. Moreover, a different sensitivity to temperature and seed osmoconditioning was ascertained in the two species. In sunflower, at 15°C, priming increased the respiration rate (+186%) compared to control just after 5 hours from initial seed imbibition; at this temperature, high respiration levels in the control as well (up -1 to 12 micromoles 10-3 CO2 s 10 g of dry seeds) where recorded. In sorghum, a positive response to osmopriming was observed in terms of seed respiration, but after 5 hours this last was the 90% higher than the control one; also, a maximum value of 6 micromoles at 15°C for unprimed seeds was achieved, showing a higher sensitivity of this species to suboptimal temperature compared to sunflower.

234 - SEED MOISTURE DISTRIBUTION IN A SOYBEAN PLANT PESKE, SILMAR T., HÖFS, ALBERTO & HAMER, ELTON

Federal University of Pelotas, C.P.354, Pelotas RS, Brazil, [email protected]

It is common to see in any soybean plant, that seeds reach maturity at different times. So, the objective of the present study was to determine the magnitude of the seed moisture distribution at different stages of maturation in a soybean plant. The field study was conducted in a tropical region in the state of Mato Grosso – Brazil. The field was established with foundation seeds of MTBR-45 cultivar, and at flowering, 100 plants were marked at same maturity stage. Harvesting began when seeds had still high moisture content where, at each time during 16 days, 100 pods with two seeds, from two plants, were harvested. These seeds from each pod were hand treshed individually and determined the moisture content. The results revealed that there is a great distribution of seed moisture content in a soybean plant, where at field maturity, the magnitude can reach more than 10 percentage points. Also, in order to have all seeds with safe moisture content for storage, an high percentage of seeds will be waiting for more than four days to be harvested. It is recommended that harvesting be accomplished when seed MC content be 15-18% in order to minimize field deterioration and, artificial drying, even when the seeds have low MC, due to their high moisture range.

235 - SEED DEVELOPMENT IN HIMALAYAN MAPLE (ACER CAESIUM) PHARTYAL* S.S. & THAPLIYAL, R.C.

Forest Tree Seed Laboratory, Forest Research Institute, DehraDun (UA) INDIA

Seed development in Acer caesium was studied from anthesis to maturation during the year 1997 and 1998 at a site in the western Himalayas in India. Seeds matured in 210-225DAA (days after anthesis). There were slight differences in fruit and seed constituents in two

144 Budapest, Hungary, May 17th – 19th, 2004 different years of study. With the progress of maturity, pericarp and testa colours of seed turned dark brown. This stage coincided with maximum dry matter, germination per cent, minimum fresh weight and moisture content (MC). MC in all parts of samara showed an abrupt decline during the final phase (210-225DAA) of maturation and stabilized around 12- 14% in mature seed. Changes in both calcium and potassium were not very consistent while the sugar content declined and remained relatively at low concentration during the entire course of seed development. Contrary to this, starch content first decreased upto 75DAA then increased sharply till 195-210DAA. Similar increasing trends are shown by the protein content, which reach peak when seed undergoes a desiccation phase with the maturity. The study further revealed that A. caesium seed underwent a triphasic development pattern after anthesis.

236 - PHYSIOLOGICAL CHARACTERS OF WELSH ONION SEEDS (ALLIUM FISTULOSUM L.) DURING REDUCING OF GERMINATION PERCENTAGE AND ITS HORMONE REGULATION QINGQUAN, S., RONGQI, G. & YANPING, Y.

Seed Science and engineering department, Agronomy Academy, Shandong Agricultural University, Taian, Shandong Province, 271018, P. R. China

Seed germination percentage (GP) is nearly correlative with the content of iPAs (Isopentenyladenosine Group) and ABA in Welsh onion (Allium fistulosum L.) seeds. The content of iPAs of Welsh onion seeds reduces with the reducing of seed GP, but the content of ABA of Welsh onion seeds increases with reducing of seed GP. With the reducing of GP of Welsh onion seeds, damnification degree of cell membrane becomes ingravescence, it increases that the content of amino acid and soluble sugar and K+ thickness and content of amino acid in seeds is, but it decreases obviously that the fresh weight of a seedling and the vigor index and the value of pH. The effect of hormone regulation is different from high to low GP of seeds. Being treated for 14h according to “1mg/l KT+1mg/l GA3”, hormone regulation could have notability effect for the seeds (83.5%>GP>61.5%), most notability effect for the seed (18.5%>GP>11.5%), and no notability effect for the dead seeds (GP=0%).

237 - MONITORING THE DETERIORATION PROCESS IN CORN SEEDS RUBENS SADER 1, SIMONE APARECIDA FESSEL 2 e ÉDILA VILELA DE RESENDE VON PINHO 3

1Crop Science Dept. – College of Agrarian Sciences. Campus of Jaboticabal UNESP UNIVERSITY. Zip Code: 14884-900. Jaboticabal, São Paulo State (SP), Brazil, [email protected] 2Seed Production and Technology FCAV/UNESP – Campus of Jaboticabal, SP, [email protected] 3Federal LavrasUniversity (UFLA), Lavras, Agriculture Dept., Minas Gerais State, Brazil, [email protected]

This research was developed with the objective of monitoring the deterioration process in seeds and embrionary corn tissues. It was used an hybrid corn seed lot 9569 from the Dow AgroSciences Seed Company. The seeds were evaluated in relation to it’s water content, standard germination test and seed vigor (electrical conductivity, accelerated aging and TZ) and field emergence. The enzymatic activity was tested in the enzymes: alcohol dehydrogenase, catalase, transaminase oxalacetate, glutamate dehydrogenase and peroxidase. For the enzymes extraction was selected at random two hundred seeds sectioned and divided in the three parts: coleoptile-plumule; seminal-roots and, the mesocotyl and the third one’s composed by the coleorhiza, radicle, including in all the parts the scutellum and the endosperm. Was observed higher activity of the enzymes in the seminal roots and the mesocotyl and in the radicle- coleorhiza, and was concluded that the corn seeds start to deteriorate firstly in these regions.

145 27th ISTA Congress – Seed Symposium

238 - THE BIOCHEMICAL ASPECT OF MASS, PHOSPHORUS AND WATER CONTENT IN SOYBEAN SEEDLINGS. RELATION TO THEIR CONTENT IN SEED SREDOJEVIĆ S.1, SREBRIĆ M.1, DJUKANOVIĆ L.1, PIPER P.1, VRVIĆ M.2, KOLAR-ANIĆ L.J.3

1Maize Research Institute "Zemun Polje", Slobodana Bajića 1, 11185 Zemun Polje, Serbia and Montenegro 2Faculty of Chemistry, Studentski trg 6, 11000 Belgrade, Serbia and Montenegro 3Faculty of Physical Chemistry, Studentski trg 6, 11000 Belgrade, Serbia and Montenegro

The growth of seedlings and plants of four soy bean sorts depended of seed mass and phosphorus in it, than of water absorption and soil sorts. The water income had diluted the seed substance from 9000 gL-1 down to the 154 gL-1. Dilution was the highest in sort with highest input of water but the highest concentration was finding in the sort with the greatest seed mass. The mass and phosphorus in relation seed: seedling expressed release of substance of about 20 % in examined seven days old seedlings. Opposite to the mass release, of phosphorus was with bright range without any regularity connected to sorts or substrates. Further more the same parameters in the relation seedling: plant was shown that only 1/4 to 1/3 of substance enter to the plant. The grater share of phosphorus was detected in plant grown on substratum compared to the same in control. It could be described to superset P in plant, or to slight influx independently of soil type. According to the ratio of water in seedling plant, both the substance and water were directed to plant in shernozem; water in eutric cambisol and substance in pseudogley.

239 - EFFECTS OF DIFFERENT DRYING RATES ON THE VIABILITY OF SEEDS OF COFFEA CANEPHORA PIERRE. STTELA DELLYZETE VEIGA FRANCO DA ROSA1, DELACYR DA SILVA BRANDÃO JÚNIOR2, ÉDILA VILELA DE R. VON PINHO3, ANDRÉ DELLY VEIGA4, LUIZ HILDEBRANDO DE C. SILVA

1Embrapa Café, Cepecafé, CP 37, Campus Ufla/Dag, Cep 37200-000, Lavras, MG, [email protected]; 2-Pesquisador Dr., FAPEMIG/Ufla/Dag, CP 37, Cep 37.200-000, Lavras, MG; 3Ufla/Dag, CP 37, Cep 37.200-000, Lavras, [email protected]; 4Mestrando do curso de Fitotecnia da Ufla/Dag, Lavras, MG, [email protected]

Desiccation tolerance of seeds only depends upon characteristics inherent to the species, of the developmental stage and of the conditions under which the seeds are dried, particularly upon the velocity which water is removed from the seeds. Slow drying may induce tolerance to desiccation in orthodox seeds but in contrast in recalcitrant seeds results into less tolerance, being that the faster dehydration may be achieved, less is water content in which seeds or embryonic axes may be dried without losing viability. Coffea seeds are regarded as of intermediary behavior for they tolerate a relative dehydration when compared with those orthodox and re sensitive to low temperatures. The objective of this work was to verify the effect of different drying rates on the viability and the storability of Coffea canefora Pierre seeds. The experimental design was completely randomized, a 3 x 3 factorial scheme with three drying rates (fast, intermediary and slow), five final water contents of drying (50, 40, 30, 20 and 15% bu) and two storage temperatures under hermetic conditions (10 and 20ºC). Germination and vigor evaluations by the root protrusion tests at fifteen days of germination of cotiledonary leaves at 45 days of germination, of emergence velocity index, were accomplished soon after drying treatments, at two and four months of storage. It was found that with reducing water content of the seeds occurred the reduction of the values of germination and vigor of seeds, for all the drying rates, the highest reductions in physiological quality occurred when seeds were dried fast and the best results did with the intermediary drying rate. There was an effect of drying and storage temperature on the physiologic quality of seeds, the lowest values of germination and vigor were observed on seeds with the poorest water contents and stored at 20ºC. Coffea canephora seeds were tolerant to desiccation up to 15% bu of water content and may be stored for four months under hermetic conditions and under temperature of 10ºC.

146 Budapest, Hungary, May 17th – 19th, 2004

240 - EXOGEN CAFFEINE INHIBITS THE IN VITRO DEVELOPMENT OF EMBRYOS OF COFFEA ARABICA L. AND COFFEA CANEPHORA PIERRE.

STTELA D. VEIGA FRANCO DA ROSA1, CÍNTIA GUIMARÃES DOS SANTOS2, RENATO PAIVA3; RENATO MENDES GUIMARÃES3, LEONARDO QUEIROZ DE MELO4, ANDRÉ DELLY VEIGA5.

1Embrapa Café, Cepecafé, CP 37, Campus Ufla/Dag, Cep 37200-000, Lavras, MG, [email protected]; 2Doutoranda curso de Fisiologia Ufla/Dbio, CP 37, Cep 37.200-000, Lavras, MG, [email protected] ; 3Ufla/Dag, CP 37, Cep 37.200-000, Lavras; 4Engenheiro Agrônomo, Departamento Técnico da Agrichem do Brasil, [email protected]; 5Mestrando do curso de Fitotecnia da Ufla/Dag, Lavras, MG, [email protected]

The slow germination of coffee seeds remains non-clarified though it is always stressed in studies on physiologic aspects of this species. The presence of the endocarp poor water and O2 uptake, the presence of natural inhibitors or hormonal balance have been suggested. Caffeine, an alcholoid present in coffee seeds amounts to 1 to 2% of the dry weight of the seed or a mean of 40mM and may inhibit seed germination or seedling growth but studies of germination of coffee seeds by action of endogen and/or exogen caffeine are scarce. The present work was intended to investigate the effect of exogen caffeine upon germination and development of embryos of Coffea arabica L. and of Coffea canephora Pierre. The experiment was conducted in the Laboratório de Cultura de Tecidos/Fisiologia Vegetal/DB of the UFLA, by utilizing berries at the red ripe stage of the cultivars Rubi and Apoatã IAC-2258. After desinsfetation of the berries for 30 minutes’ immersion in sodium hypochloride (20% i.a) and washing for three times in distilled and autoclaved water, the embryos were removed and inoculated in an aseptic way on Petri dishes with 50% MS medium added of sucrose and supplemented with different concentrations of caffeine (0.00; 0.5; 0.10; 0.15; 0.20; 0.25; 0.30 and 0.40%). The embryos were kept in growth room at 27+-2°C and light intensity of 13 umol.s-1.m-2 for 23 days, when shoot length, root length and seedling fresh weight were evaluated. Five days after cultivation, the percentage of emission of radicles and cotyledons, computing the embryos with open cotyledons ad with expanded radicles. The experimental design utilized was completely randomized with 6 replicates per treatment, each replicate being mad up of 5 embryos. Germination and development of embryos of Coffea arabica L. and of Coffea canephora Pierre are affected by exogen caffeine, this effect being more drastic in rootlets than in cotyledons In embryos of Coffea arabica L., caffeine concentrations superior to 0.20% drastically affected radicle emission. As opposed, rootlet emission in embryos of Coffea canephora Pierre is not affected at all up to the concentrations of 0.30%, denoting that this species is less sensitive to the effects of exogen caffeine.

241 - FORMATION OF COFFEE SEEDLINGS OF COFFEA ARABICA L.; CV RUBI BY UTILIZING SEEDS AND BERRIES AT DIFFERENT DEVELOPMENTAL STAGES

STTELA DELLYZETE VEIGA FRANCO DA ROSA1, LEONARDO Q. DE MELO2, ANDRÉ DELLY VEIGA3, SIRLEI DE OLIVEIRA3, CARLOS ALBERTO SPAGGIARI DE SOUZA4, VINÍCIUS DE ARAÚJO AGUIAR5.

1Embrapa Café, Cepecafé, CP 37, Campus Ufla/Dag, Cep 37200-000, Lavras, MG, [email protected]; 2Engenheiro Agrônomo, Departamento Técnico da Agrichem do Brasil, [email protected] 3Mestrando do curso de Fitotecnia da Ufla/Dag, Lavras, MG, [email protected]; 4Pesquisador Dr. Ceplac/Cepec/Linhares, ES, [email protected] ; 5Aluno de Agronomia/Ufla).

Coffee seeds present slow and germination and poor storage potential which makes seedling formation in a suitable time and under climatic conditions favorable to crop establishment. The propagation of the coffee tree by means of the seedlings from seeds is still widely performed and the reduction of the time for the obtaining of well developed and vigorous seedlings, aiming at the good establishment of the stand and reduction of the percentage of re-planting. Taking into account that coffee seeds acquire their maximum germination at the verde cana and cereja stages, the present work was performed with a view to testing alternative for obtaining of seedlings, by utilizing the seeds and berries at several developmental stages. The experiment was conducted in the seedling nursery of the Coffee Production Sector of the Universidade Federal de Lavras. The design was in randomized blocks with four replicates and the seedlings were produced in small bags with an earth,

147 27th ISTA Congress – Seed Symposium manure, simple phosphate and potassium chloride substrate (standard substrate). Nine no- tillage treatments were tested: 1) sowing of the berrie at the green stage ; 2) of the berrie at the green stage after 10 days from harvest; 3) of the berrie at the yellow stage;4) of the berrie at the yellow stage after days from harvest; 5) of the berrie at the red ripe stage; 6) of the seed of the red ripe, desmucilada and dried up to 15% of water content; 7) of the seed of red ripe, desmucilada, dried up to 15% without parchment 8) of the seed of red ripe, desmucilado, dried up to 15% of water content and pre-deepened into water for six days and 9) of the seed of the cherry fruit , desmucilado and dried up to 15% without parchment and pre-deepened in water for six days. To verify the effect of the treatments, after 140 day from the start of the experiment, the emergence percentage (E), the emergency velocity index (EVI) and the percentage o seedlings with at least one pair of true leaves (TL) was evaluated and after six months the evaluation of by means of seedlings was proceeded by means of measurements of stem diameter (D), plan height (H), of the dry matter of the root system (DMRS), of the dry matter of the shoot (DMS), of the leaf area (LA) and of the number of pairs of leaves (NPL). The treatments which stood out, according to the statistic analysis, were sowing of green berry, of seeds with and without a parchment and of seeds without a parchment and deepened in water for six days for the characteristics E, D DMRS and NPL. The seedlings from the sowing of seeds from berrie at the red ripe stage and without a parchment were the ones which presented the best performance of growth.

242 - GERMINABILITY AND DESICCATION TOLERANCE IN CORN SEEDS HARVESTED AT DIFFERENT MATURATION STAGES VON PINHO R.G., FARIA, M.A.V.R., VON PINHO, É.V.R., GUIMARÃES, R.M. & FREITAS, F.E.O.

Agronomy Departament / Federal University of Lavras, Brazil, zip: 37200-000, phone: 55 3538291315, [email protected]

Aiming at obtaining additional subsides to decision-making concerning harvest time and drying of corn seeds on ears, biochemical and physiological aspects of seeds harvested at different milk line stages and submitted to artificial drying was studied The experiments were accomplished at Seed Laboratory of Federal University of Lavras, Brazil. Seeds of the hybrids AG-9090, AG-9010, DKB-350 and DKB-747, produced by Monsanto do Brasil S.A in the 2000 winter in the towns of Iraí de Minas – MG, Campo Florido – MG, Miguelópolis – SP and Guaíra – SP were utilized. In each field, four rows were demarcated where 100 ears per milk line stage (namely ML2, ML3, ML4 and ML5, containing , respectively, 25%, 50%, 75% and 100% of hardened endosperm), were harvested. The ears were transported to that lab and were sampled for determination of seed water content. After straw removal, ears were taken to experimental dryers where they were dried at initial temperature of 35oC till they reached 20% of water content, followed by temperature of 42oC up to 12% of water content. Tests of germination, electric conductivity, cold test without soil and accelerated aging, electrophoresis analyses of α-amylase and heat resistant proteins were carried out, utilizing freshly harvested or dried seeds. We concluded that the germinability of corn seeds increases with water loss in the field and is fully obtained by means of a moderate drying of the seeds harvested from milk line stage 3 (ML3), phase in which seeds already achieved desiccation tolerance and present a high physiological quality.

243 - TOLERANCE OF CORN SEEDS TOWARDS HIGH DRYING TEMPERATURE: BIOCHEMICAL AND ANATOMICAL ASPECTS VON PINHO, E.V.R., ROVERI JOSÉ, S.C.B., VON PINHO, R.G. & RAMALHO, M.A.P.

Agronomy Departament / Federal University of Lavras, Brazil, zip: 37200-000, phone: 55 3538291315, [email protected]

Several mechanisms have been involved in the acquisition and maintenance of desiccation tolerance. The objective of this research was to evaluate the biochemical aspects, as the α- amylase’s activity and the electrophorectic patterns of the heat resistant proteins, and also the anatomical aspects of the seed pericarp, that could be associated with the high drying temperature tolerance. Seeds from lines, produced in 1999/2000, classified as tolerant and 148 Budapest, Hungary, May 17th – 19th, 2004 intolerant to high drying temperature, and seeds from hybrids which present significant reciprocal effect, produced in 2000/2001, were used. The seeds were harvested with 35% moisture content and dried at 45oC. Seeds dried under shadow were utilized as control in 1999/2000. The α- amylase enzyme was extracted from germinated seeds from each material in TRIS – HCl 0,2 M buffer, and the heat resistant proteins, of the embrionary axes, in TRIS – HCl 0,05 M buffer. Seeds that presented superior physiological quality, evaluated by the germination test, also presented greater band intensity for the α-amylase enzyme. A higher concentration of heat resistant proteins, of molecular weight between 18 and 44,1 kDa was verified, in seeds from the tolerant lines. The electrophorectic patterns of the heat resistant proteins were similar amid the hybrid seeds which presented significant reciprocal effect. Differences in seed tolerance to high drying temperature were related to the physical characteristics of the pericarp. Seeds from lines and hybrids that presented denser pericarp structure, formed by more compact cells, were more sensitive to high drying temperature.

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