Brief Summary of Minutes of Annual Meeting

The USDA-ARS National Seed Storage Lab and Colorado State University, Fort Collins, Colorado hosted the Year 2001 Annual Regional Technical Advisory Committee (RTAC) meeting of NE-009 on June 26, 2001. Warren Lamboy, in Kim Stoner‘s absence, chaired the business portion of the meeting. The PGRU report to the committee included discussion of preservation and research highlights, personnel changes, improvements to facilities, and future directions and plans. Guest presenters included Peter Bretting, National Program Leader from the USDA-ARS National Plant Germplasm System, Ann Marie Thro of USDA-CSREES, and David Tay and Susan Stieve of the new Ornamental Plant Germplasm Center, in Columbus, Ohio. A new budget was proposed and passed. The RTAC suggested that Warren Lamboy and Kim Stoner solicit committee members for non-Geneva locations for the 2002 RTAC meeting.

PROGRESS OF THE WORK AND PRINCIPAL ACCOMPLISHMENTS:

In the past year approximately 12200 accessions of seed propagated crops (tomato, onion, squash, radish, cabbage, cauliflower, broccoli, other cole crops, celery, ground cherry, asparagus, and buckwheat) were maintained. There were 450 plots grown for seed production and 185 planted to produce first year bulbs and plants of biennial crops that were stored for seed production in the upcoming season. 2057 seed samples of 1501 germplasm accessions were distributed to 93 qualified researchers and bona fide users worldwide. More than 90% of the samples were for onion, Brassica, and tomato. There were approximately 125 onion accessions characterized for the Crop Germplasm Committee minimal descriptor list. Digital images were produced for both foliage and bulbs for uploading to the national germplasm database, GRIN. We obtained funding for a specific cooperative agreement (SCA) for the regeneration of the short-day onions in New Mexico and will regenerate 50 accessions of short day onions that are in danger of being lost due to poor viability and low seed numbers. Duplicate accessions of tomato were identified and research has been initiated with molecular markers to determine the degree of duplication that can be eliminated from the collection. A preliminary core collection of tomato has been identified through interaction with the tomato CGC. Separate passport and inventory files for individual vegetable crops have been combined into only one passport and inventory file to facilitate the transfer to GRIN. Programs have been developed in SAS to allow automatic calculation of the status and need for regeneration of crops.

For our clonally propagated crops, 3744 accessions of apple, 1203 accessions of grape, and 103 accessions of tart cherry were maintained during the last year. In 2001, 119 orders for 1861 accessions of apple, 5 orders for 43 accessions of tart cherry, and 70 orders for 784 accessions of grape were distributed. Repropagation of the apple collection onto fire blight tolerant EMLA 7 rootstock continues and is 50% complete. It should be completed within the next 3 years. This will dramatically reduce the loss of trees from fire blight and will result in fewer CRIS resources being expended in regenerating and replacing lost trees. Evaluation by screening seedlings for disease resistance from the collection of wild apple from Turkey has begun, funded by an SCA with H. Aldwinckle. 400 of these seedlings will be field planted for horticultural evaluation. 1200, 400 and 300 seedlings, from Central Asia, China, and the Russian Caucasus, which previously screened positive for disease resistance, are in field plantings with horticultural evaluation in progress. Preliminary results indicate that both fire blight and apple scab resistance is present in the genotypes field-tested.

Nearly the entire collections of apples and tart cherries are now backed up in cryogenic storage at the USDA, ARS, National Seed Storage Lab, Fort Collins, CO. The project should be completed by 2004. This provides safe, off-site, backup storage for this germplasm. Volume 29 of "Horticultural Reviews" has been reviewed and edited and went to the publisher in Dec. 2001. It is entitled "Wild Apple and Fruit Trees of Central Asia", and comprises two books: "The Wild Apple Tree of Kazakhstan" and "Wild Fruit Plants of Kazakhstan", both translated from Russian to English as part of a PGRU SCA with Kazakhstan. Volume 29 will begin with the article "Collection, Maintenance, Characterization and Utilization of the Wild Apple of Central Asia". This is a detailed report describing the four PGRU-lead expeditions to collect wild apple in Central Asia during the years 1989-1996 and the follow-up work to evaluate and promote utilization of the genetic material.

Characterization of the grape collection for 14 fruit characteristics (weight, cluster size, etc.) is complete except for several accessions that did not fruit this past season. Leaf and flower characteristics have also been determined for 26-79% of the accessions (the % depending on the character). Tests of grape seed in long-term cold storage continued. Baseline tests on seed germination of 8 different species of fresh seed (no time spent in cold storage) resulted in 0 - 96% (ave. 52%) germination, depending on species. If cold storage is successful, the genes contained in grape accessions will be preserved in seed form as security backup. Over 400 computer images of grape clusters were taken and were put into the GRIN database for public access. Probability models predicting the number of seeds needed to preserve all the genes in a clonal crop have been developed and refinements and computer simulations are in progress.

The unit has successfully developed linkages with organic farming groups and seed savers organizations. Larry Robertson, Dave Sharman, and Phil Forsline made four presentations to the Northeast Organic Farmer-s Organization-New York annual conference in Syracuse, NY this year. PGRU will take the lead in organizing training workshops for small-scale vegetable seed production and participatory on-farm demonstrations of new publicly bred vegetable varieties with small seed companies and organic farmers as part of a successful three year project funded by the Initiative for Future Agriculture and Food Systems (IFAFS). Funding is provided for a horticulturist and the creation of a mobile seed-processing unit. PGRU will also have a major role in developing participatory plant breeding with these non-traditional stakeholders. Through this project, modern publicly bred cultivars will be identified for inclusion in the germplasm collections.

A scientist was hired to direct the activities in the PGRU Molecular Biology Lab and new equipment was purchased for carrying out medium- to high-throughput genotyping. This equipment allows the simultaneous processing of 192 plant samples, from the stage of grinding leaf tissue to final genotyping analyses. Development of a molecular marker database to serve the lab is also underway. A web site containing lab protocols and project descriptions has been developed to aid in transfer of knowledge within the PGRU lab and in order to freely share protocols with interested outside parties.

One Hobart and William Smith Colleges summer intern and two Research Apprentice Program (RAP) students were trained in the PGRU lab as Biological Science Aides in summer 2001. They assisted in optimizing molecular genetic markers in tomato and broccoli, and collecting data from pilot projects involving DNA fingerprinting of tomato and broccoli accessions. They also spent time assisting with various activities in the field and in the greenhouse.

We are carrying out molecular genotyping of broccoli accessions for a locus implicated in flower morphology. Genotype will be correlated with phenotypic measures of degree of branching in the flower head. All of the phenotypic data have been collected in the field. The molecular genotypic data is 90% complete. Two tomato DNA fingerprinting projects are also being carried out. One involves comparing genetic variation in American heirloom accessions to that found in modern day commercial hybrid varieties that are commonly planted in gardens. The other project involves comparing sets of duplicate (same or similarly named) accessions. More than 50% of the molecular data and all of the phenotypic data have been collected for the tomato projects.

USEFULNESS OF FINDINGS:

Valuable crop germplasm of clonal crops and seed crops is being maintained and distributed in accordance with international germplasm standards for viability, genetic integrity, and accurate cultivar and species identity. Training of NGO participants in characterization and regeneration techniques will lead to complementary on-farm conservation of germplasm. These activities will provide benefits in the form of useful heirloom germplasm for non-traditional stakeholders who are involved in alternative agriculture and by providing a mosaic of traditional heirlooms for public consumption.

Increased characterization of unit germplasm and the addition of digital images will increase the efficiency of use of the germplasm collections. Improvements in apple maintenance procedures using a rootstock that induces lower vigor and is resistant to fire blight will increase long-term survival of apple accessions and decrease need for regeneration. Virtually all of the apple and tart cherry collections have been backed up in cryogenic storage at Fort Collins, CO. For 50 apple accessions, this has been critical as we lost them due to heavy fire blight in the field collection but were able to rescue them from our cryogenic-storage inventory. Secure backup of grape genes should be possible if long-term grape seed storage experiments are successful.

For the molecular work on the vegetable crops, genotyping of broccoli and cauliflower may lead to marker that can be used to predict flower morphology in accessions and for marker-assisted selection. For the tomato work, if patterns of genetic variation are different between modern-day versus heirloom types, the argument for characterizing and using heirlooms is made stronger because there may be valuable, underutilized genetic variation that has been unintentionally eliminated from the modern cultivars. Identification of true duplicate accessions in tomato based on phenotypic and genotypic data will aid the curator in making decisions about eliminating them, freeing up resources currently used to maintain them


WORK PLANNED FOR NEXT YEAR

Regenerations of cabbage, cauliflower, and broccoli will continue, and 25 accessions of Cucurbita are scheduled for regeneration. Fifty accessions of cabbage grown last year will be planted for seed production and another 60 will be grown for first year plants for regeneration of seed in this and each subsequent year for the next nine years. Regenerations will continue for the other Brassica oleracea and Brassica rapa varieties that have low seed supply. Bulbs from approximately 200 accessions of long-day onion will be planted for seed production at Geneva and by private seed company collaborators. First-year bulbs will be produced for the remaining long-day onion accessions that need regeneration and have seed available. We are attempting to identify funding for an SCA for the regeneration of the short-day onions in New Mexico. SunSeeds will regenerate 12 short-day onion accessions in southern California. Through collaboration with New Mexico State University regeneration of 50 short day onions in critical need for rejuvenation will be initiated with the production of first year bulbs. Five hundred tomato accessions that do not have data for the minimum descriptor list will be planted for characterization. The switch to exclusive use of GRIN for our vegetable seed collections should be completed and the local databases will be !%locked!&. As part of the IFAFS collaborative project with Cornell University on-farm conservation and participatory plant breeding workshops will be developed and conducted in collaboration with the other partners of the project (NOFA-NY and the Farmers Cooperative Genome Project). Additionally, on-farm demonstrations of new publicly bred vegetable varieties will be initiated through this project and a database of public vegetable breeders and varieties will be developed.

The sour cherry collection with the initial field planting in spring 1998 is fruiting for the first time in summer 2001. By July of 2002, we can begin characterizing for minimal descriptors those accessions for which we lack this information. This data will be added to data already in GRIN that was obtained in 1999 from Dr. Amy Iezonni of Michigan State University. Forty accessions from her field plantings were characterized in a study done in East Lansing, MI.

More than eight races of apple scab (Venturia inaequalis) have been identified and breeders have been unable to develop durable sources of apple scab resistant cultivars. In collaborative work with New Zealand scientists and with H. Aldwinckle at Cornell University we have identified sources of resistance in the Central Asian germplasm that has potential as new parental material in a breeding program. In spring 2002, PGRU will make crosses of scab resistant selections of Malus sieversii and a susceptible cultivar, ‘Gala‘. Seeds from these crosses will be screened at Cornell and in New Zealand. Using genetic markers, these populations and a broad spectrum of Malus sieversii will be characterized for the different scab resistant genes for which markers have been developed. This work will be expanded to include the laboratory of J. Luby at the University of Minnesota.

Repropagation of the apple collection onto fireblight resistant rootstocks will continue. Characterization of the grape collection for leaf descriptors will continue, and apple characterization will continue to include digital imagining of fruit and tree morphology. Over 3000 seedlings of Malus species are also being evaluated for horticultural traits after being screened for resistance to three apple diseases. Additional work in grape genomics or taxonomy is expected for 2002, but specific activities have not yet been decided upon.

The lab work on the vegetable crops will continue to focus on species that are a high priority with respect to characterization (tomato and Brassicas). Optimized molecular markers will be applied to these species, and molecular marker data combined with phenotypic data will be analyzed and reported. We will continue to adopt and optimize new marker systems for tomato and Brassicas, continue to develop a molecular marker database for the lab, and update and maintain the lab web site.

Aldwinckle, H.S., P.L. Forsline, H.L Gustafson and M.V.B Reddy. 2001. Fire blight resistance Malus species from Sichuan (China), Russian Caucasus, Turkey and Germany. The 9th International Workshop on Fire Blight, New Zealand. Acta Horticulturae (in press).

Benson, L.L., R.H. Zimmerman, and W.F. Lamboy. 2001. Molecular Identification of Malus hupehensis (Tea Crabapple) Accessions Using Simple Sequence Repeats. HortScience 36: 961-966.

B|ttner, R., M. Fischer, P.L. Forsline, M. Geibel and V.V. Ponomarenko. 2001. Genebank work for preservation of the genetic diversity of apple. EUCARPIA XVIth Genetic Resources Section Symposium, !‘Broad Variation and Precise Characterization - Limitation for the Future!(, Poznan, Poland, May 16-20, 2001.

Cantini, C., A.F. Iezzoni, and W.F. Lamboy. 2001. DNA Fingerprinting of Tetraploid Cherry Germplasm Using Simple Sequence Repeats. Journal of the American Society of Horticultural Science 126: 205-209.

Forsline, P.L. and H.S. Aldwinckle. 2001. Natural occurrence of fire blight in USDA Apple collection after 10 years of observation. The 9th International Workshop on Fire Blight, New Zealand. Acta Horticulturae (in press).

Forsline, P.L. H.S. Aldwinckle, E.E. Dickson, J.J. Luby, S.C. Hokanson, G. Mink, L.L. Benson and A.D. Djangaliev. 2001. Wild Malus, Apple collections from Centers of Origin in Central Asia and Beyond. Symposium on: !‘Plant Germplasm Exploration--Adding to our Global Treasures!(. CD-ROM, ASA, CSSA, SSA Annual Meeting Abstracts, October 21-25, Charlotte, NC.

Hannan, R., N. Acikgoz and L.D. Robertson. 2001. Chapter 6: Chickpeas (Cicer L.). p. 115-124 In: N. Maxted and S.J. Bennett, Plant Genetic Resources of the Mediterranean. Kluwer Academic Publishers, Dordrecht, The Netherlands.

Hokanson, S.C., W.F. Lamboy, A.K. Szewc-McFadden, and J.R. McFerson. 2001. Microsatellite (SSR) variation in a collection of Malus species and hybrids. Euphytica.118: 281-294.

Luby, J., P.L. Forsline, H.S. Aldwinckle, V. Bus and M. Geibel. 2001. Silk Road Apples !V Collection, Evaluation, and Utilization of Malus sieversii from Central Asia. Proceedings paper from Workshop 11 !‘Collection, Utilization, and Preservation of Fruit Crop Genetic Resources-Some case studies!(. HortScience 36:225-231.

Pavek, D.S., W.F. Lamboy, and E.J. Garvey. 2001. Selecting In Situ Conservation Sites for Grape Genetic Resources in the U.S.A. Accepted in Genetic Resources and Crop Evolution.

Robertson L.D. and N. Maxted. 2001. Chapter 14: Ex situ conservation of grain legumes. p. 245-262 In: N. Maxted and S.J. Bennett, Plant Genetic Resources of the Mediterranean. Kluwer Academic Publishers, Dordrecht, The Netherlands.

Street, K.A., L.D. Robertson and N. Maxted. 2001. Chapter 18: Utilisation of pasture legume diversity. p. 327-355 In: N. Maxted and S.J. Bennett, Plant Genetic Resources of the Mediterranean. Kluwer Academic Publishers, Dordrecht, The Netherlands.

Wouw, M. van de, D. Enneking, L.D. Robertson and N. Maxted. 2001. Chapter 8: Vetches (Vicia L.). p. 134-158 In: N. Maxted and S.J. Bennett, Plant Genetic Resources of the Mediterranean. Kluwer Academic Publishers, Dordrecht, The Netherlands.