Progress 01/29/03 to 01/28/08
Outputs
Progress Report Objectives (from AD-416) To improve, expand and refine the program to efficiently and effectively conserve germplasm of Medicago, Trifolium (perennial) and Lotus for the National Plant Germplasm system. To support, where relevant and feasible, germplasm evaluation projects. To provide comprehensive and accurate information as well as in order to promote the use of these collection. Approach (from AD-416) Eliminate accession duplication using established procedures and classify the remaining accessions in order to quantify germplasm sample quality. Develop guidelines for the future expansion of the collections by assessing the existing collection using novel scientific approaches. Fill gaps in the genetic base of the collections through plant collection or exchange activities. Regeneration of the forage legume species will be accomplished under controled pollination at Prosser, WA. Facilitate the use of the collection by linking information sources, improving the quality and quantity of information and providing guidance in accession selection by users. REPLACING 5348-21000-015-00D(01/03). Significant Activities that Support Special Target Populations This progress report addresses the work conducted by the National Temperate Forage Legume Germplasm Resources Unit in Prosser, WA. The project focuses on the problem of effectively maintaining and charactering a large germplasm collection in order to enhance the use and conservation of these important genetic resources and aligns with National Program 301 Action Plan Component 1: Plant and Microbial Genetic Resource Management. Problem Statement 1A: Efficiently and Effectively Manage Plant and Microbial Genetic Resources. This project is being terminated and was replaced by 5348-21000-022-00D in January, 2008. Please refer to the report for 5348-21000-022-00D for more information. Technology Transfer Number of Other Technology Transfer: 1
Impacts
(N/A)
Publications
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Progress 10/01/06 to 09/30/07
Outputs
Progress Report Objectives (from AD-416) To improve, expand and refine the program to efficiently and effectively conserve germplasm of Medicago, Trifolium (perennial) and Lotus for the National Plant Germplasm system. To support, where relevant and feasible, germplasm evaluation projects. To provide comprehensive and accurate information as well as in order to promote the use of these collection. Approach (from AD-416) Eliminate accession duplication using established procedures and classify the remaining accessions in order to quantify germplasm sample quality. Develop guidelines for the future expansion of the collections by assessing the existing collection using novel scientific approaches. Fill gaps in the genetic base of the collections through plant collection or exchange activities. Regeneration of the forage legume species will be accomplished under controled pollination at Prosser, WA. Facilitate the use of the collection by linking information sources, improving the quality and quantity of information and providing guidance in accession selection by users. REPLACING 5348-21000-015-00D(01/03). Accomplishments Maintained, characterized and distributed forage legume germplasm. Increased 350 accessions and completed germination tests on 400 accessions. Obtained digital images and uploaded into GRIN, 1100 images of seeds and flowers regenerated over the past 3 years. Distributed 228 seed packets of Lotus, representing 34 different species; 2236 seed packets of requests for Medicago, representing 30 different species, and 461requests for Trifolium, representing 26 different species. This accomplishment addresses the problem of effectively maintaining and charactering a large germplasm collection in order to enhance the use and conservation of these important genetic resources. This work was done at the National Temperate Forage Legume Germplasm Resources Unit in Prosser, WA. This accomplishment aligns with NP 301 Action Plan Component 1: Plant and Microbial Genetic Resource Management. Problem Statement 1A: Efficiently and Effectively Manage Plant and Microbial Genetic Resources.
Impacts
(N/A)
Publications
- Nair, R., Hughes, S.J., Ellwood, S., Oliver, R., Greene, S.L., Delalande, M., Wen, J., Oldroyd, G. 2006. Medicago truncatula Stock Center. In: Medicago Truncatula Handbook. Available: http://www.noble. org/MedicagoHandbook
- Greene, S.L., Hughes, S.J., Nair, R., Huguet, T., Aouani, M.E., Prosperi, J.M., Delalande, M. 2006. Wild Accessions and Populations. In: Medicago Truncatula Handbook. Available: http://www.noble.org/MedicagoHandbook
- Greene, S.L., Minoura, T., Steiner, J.J., Pentecost, C.G. 2007. Webgrms: prototype software for web-based mapping of biological collections. Biodiversity and Conservation Journal 16:2611-2625.
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Progress 10/01/05 to 09/30/06
Outputs
Progress Report 1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? Why does it matter? This project is aligned with National Program 301. A survey conducted by the NTFLGRU among 60 ex situ collections of Medicago and Trifolium, indicated that forage legume scientists/curators around the world consider that genetic erosion is occurring in these genera. This reflects the serious problem of conserving forage legume genetic resources before they disappear. Of concern, wild relatives of our important forage legume crops are poorly represented in global ex situ collections. A broad and diverse genetic base, especially of species closely related to cultivated forages is critical for the development of new cultivars and the improvement of existing cultivars of forage crop species. Ex situ conservation and ready availability of diverse collections of genes is an efficient way to
ensure the conservation and promote the use of broad- based forage legume genetic resources and reflects the primary goals of this project. Specifically we are addressing the problems of conserving and using genetic resources by i) acquiring and conserving Medicago, Trifolium, and Lotus genetic resources and associated information; ii) characterizing selected germplasm, using morphological and molecular marker technology and incorporating the resulting data into publically accessible databases; iii) assessing whether selected wild forage legume genetic resources are more effectively conserved in situ or ex situ; iv) transferring technology in the form of forage legume genetic resources and associated information to researchers and breeders worldwide. The results not only provide genetic resources and information to end users, but also assist decision-making between various management goals that optimize the conservation of genetic resources. The interdependence of project objectives
for meeting conservation and end user goals, reflect the complexity and multi-phase nature of genetic resource management. 2. List by year the currently approved milestones (indicators of research progress) Year 1 (FY04) Germplasm collecting (native species) Germplasm collecting (Medicago truncatula) Ex situ collection regeneration, regeneration of 400 accessions Characterization during regeneration 400 accessions, completed taxonomy verification Alfalfa duplication study, completed field evaluation- morphological and agronomic; Medicago truncatula evaluation, completed field evaluation; completed preliminary data analysis, reported results at NAAIC conference. Completion of collection evaluation Ex situ, continued development of ex situ seed increase protocols In situ, completed species inventory and recollected seed (June, July 2004). Developed GIS database Year 2 (FY05) Germplasm collecting (native species): enter data into GRIN Germplasm collecting (Medicago truncatula): Prepare
exploration proposal Ex situ collection regeneration: Completed regeneration of 400 accessions Characterization during regeneration: Completed taxonomy verification on 400 accessions; photo document 250 accessions, obtained plant habit and flower color data Alfalfa duplication study: Analyze, summarize and interpret data Medicago truncatula evaluation: Complete data analysis, upload data into GRIN. Continuation of collection evaluation. Ex situ conservation of wild species: continued development of ex situ seed increase protocols. In situ- conservation wild species: Distribute GIS database on T. thompsonii. Locate in situ seed collection sites for other wild western US species. Year 3 (FY06) Complete regeneration of 400 accessions in ex situ collection Regenerate native species collected in 2004, newly acquired germplasm acquired in 2005 Obtain M. truncatula from other world collections Photo document 250 accessions Collaborate with R.C. Johnson to finish alfalfa duplication project
Complete upload into GRIN of M. truncatula data collected in 2003 Carry out germplasm evaluations in line with available resources and collaborators Population analysis of T. thompsonii using microsatellites Continue development of ex situ seed increase protocols Identify in situ collecting sites for T. plumosum Year 4 (FY07) Complete regeneration of 400 accessions in ex situ collection Regenerate new accessions acquired in 2006 Reassess need to collect M. truncatula and write proposal if warranted. Photo document 250 accessions Collaborate with R.C. Johnson to complete manuscript Distribute core subset for M. truncatula Carry out germplasm evaluations in line with available resources and collaborators. Population analysis of T. thompsonii using microsatellites Continue development of ex situ seed increase protocols Identify in situ collecting sites for western North American wild Lotus species. Year 5 (FY08) Complete regeneration of 400 accessions in ex situ collection Regenerate
newly acquired accessions collected in 2007 Collect M. truncatula if trip is warranted and funded. Photo document 250 accessions Complete alfalfa duplication project Complete upload into GRIN of any evaluation data collected in 2006. Carry out germplasm evaluations in line with available resources and collaborators. Complete study of T. thompsonii Continue development of ex situ seed increase protocols Identify in situ collecting sites for western species of Trifolium and Lotus 4a List the single most significant research accomplishment during FY 2006. Maintained, characterized and distributed forage legume germplasm. Increased 420 accessions; completed germination tests and assigned PI numbers for 42 G numbers and 308 W6 numbers that had been increased in the past 4 years. Photographed 350 accessions to document flowers and seeds. Distributed 10 requests for Lotus, which totaled 83 packets, 69 requests for Medicago, which totaled 2653 packets, and 32 requests for Trifolium, which
totaled 435 packets. Effective maintenance and characterization enhance the use and conservation of these important genetic resources, as evidenced by our distribution levels. This accomplishment aligns with NP 301 Action Plan Component 1: Plant and Microbial Genetic Resource Management. Problem Statement 1A: Efficiently and Effectively Manage Plant and Microbial Genetic Resources. 4b List other significant research accomplishment(s), if any. Forage legume germplasm bioinformation published on GRIN. Stephanie Greene uploaded the upgraded Lotus passport data into GRIN, as well as made numerous corrections to GRIN to improve documentation. All G numbers have had extensive documentation added to GRIN. We uploaded passport data for explorations carried out in Georgia, and the Western US in 2004 (for all species collected). Have uploaded evaluation data generated on the M. truncatula collection in a 2003 morphological evaluation, and an aluminum tolerance evaluation carried out by the
Noble Foundation. This accomplishment aligns with NP 301 Action Plan Component 1: Plant and Microbial Genetic Resource Management; Problem Statement 1A: Efficiently and Effectively Manage Plant and Microbial Genetic Resources. Improved pollination of clover germplasm. Stephanie Greene conducted an experiment to compare pollinators on 4 species of clover, (repens, pratense, fragiferum and hybridum). We found that leaf cutters set seed on 3 out of 4 species. This finding will have a significant impact on our pollinator budget, saving us at least $ 60/accession. Since on average we increase between 75-100 accessions of clovers, replacing honey bees with leaf cutters could save $ 6000 annually. Improved efficiency and use of resources resulted in improved regeneration of these forage clovers. This accomplishment aligns with NP 301 Action Plan Component 1: Plant and Microbial Genetic Resource Management; Problem Statement 1A: Efficiently and Effectively Manage Plant and Microbial
Genetic Resources. Published forage legume germplasm bioinformation on GRIN. Uploaded upgraded Lotus passport data into GRIN, as well as made numerous corrections to GRIN to improve documentation. We uploaded passport data for explorations carried out in Georgia, and the Western US in 2004 (for all species collected). Have uploaded evaluation data generated on the M. truncatula collection in a 2003 morphological evaluation, and an aluminum tolerance evaluation carried out by the Noble Foundation. Making collection information easily obtainable on the public GRIN web site will ensure the collections are effectively used. This accomplishment aligns with NP 301 Action Plan Component 1: Plant and Microbial Genetic Resource Management; Problem Statement 1A: Efficiently and Effectively Manage Plant and Microbial Genetic Resources. Reduced pollination costs in clover germplasm. Conducted an experiment to compare pollinators on 4 species of clover, (repens, pratense, fragiferum and hybridum).
We found that leaf cutters set seed on 3 out of 4 species. This finding will have a significant impact on our pollinator budget, saving us at least $60/accession; resulting in a savings of $ 6000 annually. Improved efficiency resulted in more cost effective regeneration of forage clovers. This accomplishment aligns with NP 301 Action Plan Component 1: Plant and Microbial Genetic Resource Management; Problem Statement 1A: Efficiently and Effectively Manage Plant and Microbial Genetic Resources. 5. Describe the major accomplishments to date and their predicted or actual impact. Since the project was initiated in 01/29/2003, we have regenerated 1281 accessions using state of the art seed increase procedures, distributed 11,115 packets, and entered characterization data into GRIN (4,905). We have also acquired new germplasm from the western United States, and have identified two sites for in situ seed collection. A major evaluation of the NPGS M. truncatula collection, a model species
for legume genomics, was completed. In 2004 and 2005 we began distributing this data and loaded the data into GRIN. We have also done substantial work on understanding the population dynamics of Trifolium thompsonii. Cost- effective pollination of clover germplasm was accomplished through the change to leaf cutters. Our activities have focused on ensuring the USDA germplasm collections of Medicago trifolium and Lotus continue to meet the needs of a broad array of users and contribute to the global conservation of these important genetic resources. This projects accomplishments to date address National Program 301 Component 1, Problem 1A: Efficiently and effectively manage plant genetic resources and Component 2, Problem 2A Genome database stewardship. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to
the adoption and durability of the technology products? Seed from the Lotus, Medicago and Trifolium germplasm collections was distributed to numerous users, both domestic and internationally. Compared to 2004, requests increased by 33%.
Impacts
(N/A)
Publications
- Greene, S.L., Hannan, R.M., Afonin, A., Dzyubenko, N.I., Khusainov, A. 2005. Collecting wild crop relatives in the northwestern steppes of kazakhstan. Plant Genetic Resources Newsletter. 141:1-6.
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Progress 10/01/04 to 09/30/05
Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? A survey conducted by the NTFLGRU among 60 ex situ collections of Medicago and Trifolium, indicated that forage legume scientists/curators around the world consider that genetic erosion is occurring in these genera. This reflects the serious problem of conserving forage legume genetic resources before they disappear. Of concern, wild relatives of our important forage legume crops are poorly represented in global ex situ collections. A broad and diverse genetic base, especially of species closely related to cultivated forages is critical for the development of new cultivars and the improvement of existing cultivars of forage crop species. Ex situ conservation and ready availability of diverse collections of genes is an efficient way to ensure the conservation and promote the use of broad-based forage
legume genetic resources and reflects the primary goals of this project. Specifically we are addressing the problems of conserving and using genetic resources by i) acquiring and conserving Medicago, Trifolium, and Lotus genetic resources and associated information; ii) characterizing selected germplasm, using morphological and molecular marker technology and incorporating the resulting data into publically accessible databases; iii) assessing whether selected wild forage legume genetic resources are more effectively conserved in situ or ex situ; iv) transferring technology in the form of forage legume genetic resources and associated information to researchers and breeders worldwide. The results not only provide genetic resources and information to end users, but also assist decision-making between various management goals that optimize the conservation of genetic resources. The interdependence of project objectives for meeting conservation and end user goals, reflect the complexity
and multi-phase nature of genetic resource management. 2. List the milestones (indicators of progress) from your Project Plan. Objective 1 Acquire and conserve Medicago, Trifolium, and Lotus genetic resources and associated information. Milestones: Germplasm collecting (native species)- 18 mo: write proposal; 36 mo: Carry out collection trip, 48 mo: Seed increase, passport data in GRIN, 60 mo: Seed available for distribution Germplasm collecting (Medicago truncatula)- 18 mo- Assess gaps in world collections, identify collaborators; 36 mo: Write proposal 48 mo: Carry out collection trip; 60 mo: Seed increase, passport data in GRIN Ex situ collection regeneration- 18 mo-400 accessions (350 routine field, 50 seed rescue); 36 mo: 400 accessions (350 routine field, 50 seed rescue) 48 mo: 400 accessions (350 routine field, 50 seed rescue); 60 mo: 400 accessions (350 routine field, 50 seed rescue) Objective 2. Characterize selected germplasm, using morphological and molecular marker
technology to augment conservation management, and increase utilization of germplasm collections. Incorporate the resulting data into publically accessible databases. Milestones: Characterization during regeneration- 18 mo: 400 accessions; 36 mo: 400 accessions; 48 mo: 400 accessions; 60 mo: 400 accessions Alfalfa duplication study- 18 mo: Field evaluation- morphological and agronomic; 36 mo: Analyze, summarize and interpret data; 48 mo: Apply results to other suspect duplicates in Temperate Forage Legume collections; 60 mo: Apply results to other suspect duplicates in Temperate Forage Legume collections Medicago truncatula evaluation-18 mo: Field evaluation- morphological and agronomic; 36 mo: Complete molecular marker work; 48 mo: Analyze, summarize and interpret data; 60 mo: Prepare manuscript Completion of collection evaluation- 18 mo: Identify potential collaborators, secure germplasm evaluation funding; 36 mo: Coordinate evaluations. As completed place data into GRIN; 48 mo:
Coordinate evaluations. As completed place data into GRIN; 60 mo: Coordinate evaluation. As completed place data into GRIN Objective 3 Assess whether selected wild forage legume genetic resources are most effectively conserved in situ. Milestones: Population analysis-18 mo: Produce primers, work out DNA isolation procedures; 36 mo: Complete analysis of 1995 populations; 48 mo: Complete analysis on 2004 populations; 60 mo: Complete analysis on ex situ increased populations ex situ-36 mo Complete development of ex situ seed increase protocols:; 48 mo:; 60 mo: Complete comparison of germination and storage ability of ex situ produced seed with wild collected seed in situ- 18 mo: Carry out species inventory and recollect seed (2004); 36 mo: Complete second year survey; 48 mo: Complete GIS database:; 60 mo: Complete assessment of species vulnerability using GIS analysis. Publish on status of species 3a List the milestones that were scheduled to be addressed in FY 2005. For each milestone,
indicate the status: fully met, substantially met, or not met. If not met, why. 1. Germplasm collecting (native species): enter data into GRIN Milestone Fully Met 2. Germplasm collecting (Medicago truncatula): Prepare exploration proposal Milestone Not Met Progress slowed by resource limitation (human,fiscal,equipment, etc. 3. Ex situ collection regeneration: Completed regeneration of 400 accessions Milestone Fully Met 4. Characterization during regeneration: Completed taxonomy verification on 400 accessions; photo document 250 accessions, obtained plant habit and flower color data Milestone Fully Met 5. Alfalfa duplication study: Analyze, summarize and interpret data Milestone Not Met Progress slowed by resource limitation (human,fiscal,equipment, etc. 6. Medicago truncatula evaluation: Complete data analysis, upload data into GRIN. Milestone Substantially Met 7. Continuation of collection evaluation. Milestone Not Met Progress slowed by resource limitation
(human,fiscal,equipment, etc. 8. Ex situ conservation of wild species: continued development of ex situ seed increase protocols. Milestone Fully Met 9. In situ- conservation wild species: Distribute GIS database on T. thompsonii. Locate in situ seed collection sites for other wild western US species. Milestone Substantially Met 3b List the milestones that you expect to address over the next 3 years (FY 2006, 2007, and 2008). What do you expect to accomplish, year by year, over the next 3 years under each milestone? B. Milestones expect to addressed in: FY06 Objective 1. Acquire and conserve Medicago, Trifolium, and Lotus genetic resources and associated information. Milestone: 1. Complete regeneration of 400 accessions in ex situ collection 2. Regenerate native species collected in 2004, newly acquired germplasm acquired in 2005 3. Obtain M. truncatula from other world collections Objective 2. Characterize selected germplasm, using morphological and molecular marker technology to
augment conservation management, and increase utilization of germplasm collections. Incorporate the resulting data into publically accessible databases. Milestone: 1. Photo document 250 accessions 2. Collaborate with R.C. Johnson to finish alfalfa duplication project 3. Complete upload into GRIN of M. truncatula data collected in 2003. 4. Carry out germplasm evaluations in line with available resources and collaborators. Objective 3 Assess whether selected wild forage legume genetic resources are most effectively conserved in situ. Milestone: 1. Population analysis of T. thompsonii using microsatellites 2. Continue development of ex situ seed increase protocols 3. Identify in situ collecting sites for T. plumosum FY07 Objective 1. Acquire and conserve Medicago, Trifolium, and Lotus genetic resources and associated information. Milestone: 1. Complete regeneration of 400 accessions in ex situ collection 2. Regenerate new accessions acquired in 2006 3. Reassess need to
collect M. truncatula and write proposal if warranted. Objective 2. Characterize selected germplasm, using morphological and molecular marker technology to augment conservation management, and increase utilization of germplasm collections. Incorporate the resulting data into publically accessible databases. Milestone: 1. Photo document 250 accessions 2. Collaborate with R.C. Johnson to complete manuscript 3. Distribute core subset for M. truncatula 4. Carry out germplasm evaluations in line with available resources and collaborators. Objective 3 Assess whether selected wild forage legume genetic resources are most effectively conserved in situ. Milestone: 4. Population analysis of T. thompsonii using microsatellites 5. Continue development of ex situ seed increase protocols 6. Identify in situ collecting sites for western North American wild Lotus species. FY08 Objective 1. Acquire and conserve Medicago, Trifolium, and Lotus genetic resources and associated information.
Milestone: 4. Complete regeneration of 400 accessions in ex situ collection 5. Regenerate newly acquired accessions collected in 2007 6. Collect M. truncatula if trip is warranted and funded. Objective 2. Characterize selected germplasm, using morphological and molecular marker technology to augment conservation management, and increase utilization of germplasm collections. Incorporate the resulting data into publically accessible databases. Milestone: 5. Photo document 250 accessions 6. Complete alfalfa duplication project 7. Complete upload into GRIN of any evaluation data collected in 2006. 8. Carry out germplasm evaluations in line with available resources and collaborators. Objective 3 Assess whether selected wild forage legume genetic resources are most effectively conserved in situ. Milestone: 7. Complete study of T. thompsonii 8. Continue development of ex situ seed increase protocols 9. Identify in situ collecting sites for western species of Trifolium and Lotus
4a What was the single most significant accomplishment this past year? The 12,000 accessions in the NPGS temperate forage legume collections were managed, maintained and distributed. This is important to ensure the germplasm collections continue to meet the needs of a broad array of users and contribute to the global conservation of these important genetic resources. This was done by staff at the NTFLGRU in Prosser, WA, who carried out state-of-the-art seed regeneration, producing seed on 400 accessions, including wild species and vulnerable seedlots and distributed more than 2000 seed packets. Collection management activities in 2005 will facilitate crop improvement among forage legume crops and help in conserving valuable genetic resources. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. Since the project was initiated in 01/29/2003, we have regenerated 1200 accessions using state of the art seed increase procedures. We
have also acquired new germplasm from the Western United Stated, and have identified two sites for in situ seed collection. We have accomplished a major evaluation of the M. truncatula collection. We did this research because Medicago truncatula is being used by the genomics research community as a model species for legumes. accession. In 2004 and 2005 we began distributing this data. We have also done substantial work on understanding the population dynamics of Trifolium thompsonii. Our activities have focused on ensuring theUSDA germplasm collections of Medicago, Trifolium and Lotus continue to meet the needs of a broad array of users and contribute to the global conservation of these important genetic resources. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the
technology products? Seed from the germplasm collections was distributed to numerous users, both domestic and internationally. Over 50% of the Medicago accessions requested, were for Medicago truncatula. We also provided information on the accessions to numerous users.
Impacts
(N/A)
Publications
- Kisha, T.J., Johnson, R.C., Skinner, D.Z., Greene, S.L. Variances of genetic distance measurements among alfalfa accessions compared from several molecular marker types. Crop Science Society of America. 2002. Abstract p. 175620.
- Greene, S.L. 2004. Morphological evaluation of the USDA, NPGS Medicago truncatula collection. In Proceedings of the 40th North American Alfalfa Improvement Conference, July 18-21. Quebec City, Quebec, Canada. [http://www.naaic.org/Meetings/National/2004NAAIC&TC/2004abstracts/sgreene- 2.pdf]
- Greene, S.L, T. Minoura, J. Steiner. 2004. WebGRMS Mapping software: Something new for the USDA Medicago and Trifolium collections. In Proceedings of the 40th North American Alfalfa Improvement Conference, July 18-21. Quebec City, Quebec, Canada, [Online at: http://www.naaic. org/Meetings/National/2004NAAIC&TC/2004abstracts/sgreene.pdf]
- Greene, S.L. 2003. Dealing with diversity: Seed production strategies in a forage legume germplasm collection. In Redlands Park (ed). Herbage Seeds in the New Millennium-new markets, new products, new opportunities. Proceedings of the 5th International Herbage Seed Conference. pp. 88-92. Nov. 23-26, Gatton, Australia.
- Kisha, T.J., Johnson, R.C., Skinner, D.Z., Bauchan, G.R., Greene, S.L. 2004. Variance of molecular genetic distances among alfalfa synthetic populations by marker type. Agronomy Abstracts. #5781.
- Afonin, A.N., Greene, S.L., Frolov, A.N., Dzyubenko, N.I., Levitin, M.M., Grichanov, I.Ya., Luneva, N.N., Saulich, M.I. 2005. A GIS-based interactive agricultural atlas of the Former Soviet Union with special reference to ranges of agricultural pest organisms. In: Crop Protection Conference - Management aspects of crop protection and sustainable agriculture: Research, development and information systems, p. 5-7. St. Petersburg - Pushkin, May 31-June 3, 2005, St.Petersburg - Pushkin
- Greene S.L., Afonin A.N., Frolov A.N., Dzyubenko N.I., Levitin M.M., Grichanov I.Ya., Luneva N.N, Saulich M.I, 2005. In Virtual collection of agricultural crops, their wild-growing relatives and pest organisms within the Former Soviet Union. The 5th International Symposium-Electronic biological collections: theory, standards, and perspectives of usage, Zoological Institute & Botanical Institute RAS. September, 11-18, 2005.
- Greene S.L., 2005. Collecting Legumes In The Largest Remote Region Remaining In The Lower 48 States. Aridus 17(1):1-5.
- Greene S.L., 2005. U.S. Germplasm Collection of Lotus: Activities Over the Last Decade. Lotus Newsletter, 35:109-111.
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Progress 10/01/03 to 09/30/04
Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? A survey conducted by the NTFLGRU among 60 ex situ collections of Medicago and Trifolium, indicated that forage legume scientists/curators around the world consider that genetic erosion is occurring in these genera. This reflects the serious problem of conserving forage legume genetic resources before they disappear. Of concern, wild relatives of our important forage legume crops are poorly represented in global ex situ collections. A broad and diverse genetic base, especially of species closely related to cultivated forages is critical for the development of new cultivars and the improvement of existing cultivars of forage crop species. Ex situ conservation and ready availability of diverse collections of genes is an efficient way to ensure the conservation and promote the use of broad-based forage
legume genetic resources and reflects the primary goals of this project. Specifically we are addressing the problems of conserving and using genetic resources by i) acquiring and conserving Medicago, Trifolium, and Lotus genetic resources and associated information; ii) characterizing selected germplasm, using morphological and molecular marker technology and incorporating the resulting data into publically accessible databases; iii) assessing whether selected wild forage legume genetic resources are more effectively conserved in situ or ex situ; iv) transferring technology in the form of forage legume genetic resources and associated information to researchers and breeders worldwide. The results not only provide genetic resources and information to end users, but also assist decision-making between various management goals that optimize the conservation of genetic resources. The interdependence of project objectives for meeting conservation and end user goals, reflect the complexity
and multi-phase nature of genetic resource management. 2. List the milestones (indicators of progress) from your Project Plan. Objective 1 Acquire and conserve Medicago, Trifolium, and Lotus genetic resources and associated information. Activities: Germplasm collecting (native species)- 18 mo: write proposal; 36 mo: Carry out collection trip, 48 mo: Seed increase, passport data in GRIN, 60 mo: Seed available for distribution Germplasm collecting (Medicago truncatula)- 18 mo- Assess gaps in world collections, identify collaborators; 36 mo: Write proposal 48 mo: Carry out collection trip; 60 mo: Seed increase, passport data in GRIN Ex situ collection regeneration- 18 mo-400 accessions (350 routine field, 50 seed rescue); 36 mo: 400 accessions (350 routine field, 50 seed rescue) 48 mo: 400 accessions (350 routine field, 50 seed rescue); 60 mo: 400 accessions (350 routine field, 50 seed rescue) Objective 2. Characterize selected germplasm, using morphological and molecular marker
technology to augment conservation management, and increase utilization of germplasm collections. Incorporate the resulting data into publically accessible databases. Activities: Characterization during regeneration- 18 mo: 400 accessions; 36 mo: 400 accessions; 48 mo: 400 accessions; 60 mo: 400 accessions Alfalfa duplication study- 18 mo: Field evaluation- morphological and agronomic; 36 mo: Analyze, summarize and interpret data; 48 mo: Apply results to other suspect duplicates in Temperate Forage Legume collections; 60 mo: Apply results to other suspect duplicates in Temperate Forage Legume collections Medicago truncatula evaluation-18 mo: Field evaluation- morphological and agronomic; 36 mo: Complete molecular marker work; 48 mo: Analyze, summarize and interpret data; 60 mo: Prepare manuscript Completion of collection evaluation- 18 mo: Identify potential collaborators, secure germplasm evaluation funding; 36 mo: Coordinate evaluations. As completed place data into GRIN; 48 mo:
Coordinate evaluations. As completed place data into GRIN; 60 mo: Coordinate evaluation. As completed place data into GRIN Objective 3 Assess whether selected wild forage legume genetic resources are most effectively conserved in situ. Activity Population analysis-18 mo: Produce primers, work out DNA isolation procedures; 36 mo: Complete analysis of 1995 populations; 48 mo: Complete analysis on 2004 populations; 60 mo: Complete analysis on ex situ increased populations ex situ-36 mo Complete development of ex situ seed increase protocols:; 48 mo:; 60 mo: Complete comparison of germination and storage ability of ex situ produced seed with wild collected seed in situ- 18 mo: Carry out species inventory and recollect seed (2004); 36 mo: Complete second year survey; 48 mo: Complete GIS database:; 60 mo: Complete assessment of species vulnerability using GIS analysis. Publish on status of species. 3. Milestones: A. Milestones scheduled for FY04 Objective 1. Acquire and conserve Medicago,
Trifolium, and Lotus genetic resources and associated information. -Milestones: Germplasm collecting (native species)- 18 mo: Completed proposal which was funded for 2004 collecting trip; 36 mo: Completed collection of wild legumes June, 21-24, Southeastern Oregon. -Milestones: Germplasm collecting (Medicago truncatula)- 18 mo- Completed gap assessment in world collections, identified collaborators -Milestones: Ex situ collection regeneration- 18 mo- Completed regeneration of 400 accessions Objective 2. Characterize selected germplasm, using morphological and molecular marker technology to augment conservation management, and increase utilization of germplasm collections. Incorporate the resulting data into publically accessible databases. -Milestones: Characterization during regeneration- 18 mo: 400 accessions- Completed taxonomy verification -Milestones: Alfalfa duplication study- 18 mo: Completed field evaluation- morphological and agronomic; -Milestones: Medicago truncatula
evaluation-18 mo: Completed field evaluation; Completed preliminary data analysis, reported results at NAAIC conference. -Milestone: Completion of collection evaluation- 18 mo: Led discussion to secure germplasm evaluation funding at Clover CGC (11/1/03; Led discussion to secure germplasm evaluation funding at Alfalfa CGC (7/19/04) Objective 3 Assess whether selected wild forage legume genetic resources are most effectively conserved in situ. -Milestone: ex situ-18- 36 mo Continued development of ex situ seed increase protocols -Milestone: in situ- Completed species inventory and recollected seed (June, July 2004). Developed GIS database B. List the milestones that you expect to address over the next three years. Objective 1. Acquire and conserve Medicago, Trifolium, and Lotus genetic resources and associated information. Milestones: Germplasm collecting (native species). FY05-06: enter data into GRIN, FY07-08: regenerate and make available Milestones FY05, Germplasm collecting
(Medicago truncatula). FY06: Projected Prepare exploration proposal- target exploration date: 2006; FY07: Carry out collection trip; FY08: Seed increase, passport data in GRIN Milestones: Ex situ collection regeneration. FY05-06: Complete regeneration of 400 accessions, FY07: Complete regeneration of 400 accessions,FY08: Complete regeneration of 400 accessions, Objective 2. Characterize selected germplasm, using morphological and molecular marker technology to augment conservation management, and increase utilization of germplasm collections. Incorporate the resulting data into publically accessible databases. Milestones: Characterization during regeneration- FY05-06: for 400 accessions complete plant growth habit, flower color, taxonomy verification, FY07 for 400 accessions complete plant growth habit, flower color, taxonomy verification; FY08 for 400 accessions complete plant growth habit, flower color, taxonomy verification Milestones: Alfalfa duplication; FY05: Analyze,
summarize and interpret data; FY06 - manuscript Milestones: Medicago truncatula evaluation-FY05-06: Complete data analysis, upload data into GRIN, develop of a core subset, manuscript Milestones: FY05-06: Facilitate germplasm evaluation program if funding is forth coming. Objective 3 Assess whether selected wild forage legume genetic resources are most effectively conserved in situ. Milestones: Population analysis. FY05: isolate DNA, FY06- analysis data, FY07 manuscript Milestone: ex situ-FY05-07. Continue development of ex situ seed increase protocols Milestones: in situ: FY05-07. Distribute GIS database 4. What were the most significant accomplishments this past year? A. Conservation of genetic resources is vital to U.S. agriculture. Scientists in the Plant Germplasm Introduction and Testing Research Unit in Pullman, WA, used Thompson clover as a model to determine the relative value of adopting an ex situ-only strategy versus supporting complementary in situ/ex situ strategies
for wild species. Research was done by mapping current populations of T. thompsonii, collecting seed from 4 representative populations and developing and testing an extensive GIS database and found that changes in populations occurred in last 10 years. Findings and the GIS database are available for use by the Forest Service, Washington Department of Natural Resources, and BLM, which will provide basic information needed to support the conservation of Thompson clover. B. It is important to ensure the germplasm collections continue to meet the needs of a broad array of users and contribute to the global conservation of these important genetic resources. Scientists in the Plant Germplasm Introduction and Testing Research Unit in Pullman, WA, conducted the curation of the temperate forage legume collections.The personnel at the NTFLGRU in Prosser, WA, carried out state-of-the-art seed regeneration, producing seed on 400 accessions, including wild species and vulnerable seedlots and
distributed more than 2000 seed packets. Collection management activities in 2004 will facilitate crop improvement among forage legume crops and help in conserving valuable genetic resources. Preserving the diversity of rare species and making diversity available to support research on ecological restoration using common wild species is important. Scientists in the Plant Germplasm Introduction and Testing Research Unit in Pullman, WA, collected wild and rare legume species in Southeastern Oregon.Seeds were collected at the appropriate time in collaboration with local botanists, in Oregon, Idaho and Washington. Seeds and other information are now available to scientists and restoration ecologists to conduct research. C. None D. None 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. Since the project was initiated in 01/29/2003, we have accomplished a major evaluation of the M. truncatula collection. We did this research
because Medicago truncatula is being used by the genomics research community as a model species for legumes. We planted out the entire collection of M. truncatula (319 accessions) in a replicated field trial at Prosser, WA in 2003. W collected data on 40 morphological and agronomic traits. Bulk DNA was isolated from 10 plants of 250 accessions. Results from this research will provide basic information needed to support research on functional genomics in the model species, M. truncatula. These data will not only encourage the use of the collections, but help us develop collections that broadly represent biological diversity with a manageable number of accession. In 2003 we also regenerated 400 accessions. We did this to ensure USDA germplasm collections of Medicago, Trifolium and Lotus continue to meet the needs of a broad array of users and contribute to the global conservation of these important genetic resources. 6. What science and/or technologies have been transferred and to whom?
When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Seed from the germplasm collections was distributed to numerous users, both domestic and internationally. Over 50% of the Medicago accessions requested, were for Medicago truncatula. We also provided information on the accessions to numerous users. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. Greene, S.L. Dealing with diversity: seed production strategies in a forage legum germplasm collection. International Herbage Seed Conference. 2003. Greene, S.L. Morphological evaluation of the USDA, NPGS medicago truncatula collection Alfalfa Improvement Conference Proceedings. 2003. Greene, S.L., T. Minoura, J. Steiner. Webgrms mapping software: something new for the USDA medicago and
trifolium collections. Alfalfa Improvement Conference Proceedings. 2004.
Impacts
(N/A)
Publications
- OLMSTEAD, M.A., WAMPLE, R.L., GREENE, S.L., TARARA, J.M. NON-DESTRUCTIVE MEASUREMENT OF VEGETATIVE COVER USING DIGITAL IMAGE ANALYSIS. HORTSCIENCE. 2004. V. 39(1). p. 55-59.
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Progress 10/01/02 to 09/30/03
Outputs
1. What major problem or issue is being resolved and how are you resolving it? A broad genetic base is critical for U. S. agriculture in the development of new cultivars or the improvement of existing ones in the forage crop species grown in the U. S. Ready availability of the most diverse collections of these genes is best maintained through the existing infrastructure of the USDA, ARS, National Plant Germplasm System. The fundamental problem addressed is to counteract genetic erosion in the temperate forage legume gene pool, and promote the development of genetically broad-base forage legume cultivars in the United States. These goals can only be achieved by developing efficient and effective approaches and procedures. Research is needed to test new operating procedures, develop new protocols, implement new processes and generate new data. The results not only provide genetic resources and information to end users, but also assist decision-making between various
management goals to optimize the conservation of genetic resources. The interdependence of project objectives for meeting conservation and end user goals, reflect the complexity and multi-phase nature of genetic resource management. 2. How serious is the problem? Why does it matter? The results of a 1998 survey that the NTFLGRU conducted on 60 ex situ collections of Medicago and Trifolium, indicated that forage legume scientists/curators around the world consider that genetic erosion is occurring in these genera. Importantly, the survey indicated that wild relatives of our important forage legume crops are poorly represented in global ex situ collections. The erosion of biodiversity is attributed to many forces, but importantly habitat change, which is occurring at an unprecedented rate. It is imperative to conserve forage legume genetic resources before they disappear to ensure the continued development of cultivars that support sustainable and profitable farming practices in the
United States, but also support sustainable agricultural development around the world. 3. How does it relate to the National Program(s) and National Program Component(s) to which it has been assigned? Project objectives are to safeguard and utilize a wide array of plant genetic resources for grower, processor and consumer stakeholders. In addition to directly addressing the five problem areas identified in Component I of the NP301 Action Plan, the research objectives will impact allied CRIS research projects conducted in National Programs 205, 302, 303, and 306. This project, under the MU CRIS 5348-21000-017-00D, is a vital component of the USDA, NPGS. It is responsible for the major forage legume species utilized in U.S. agriculture. Project research, information documentation and germplasm conservation relate directly to all aspects of the NP Action Plan by supporting crop development, helping to sustain small farm agriculture, preserving endangered species, repatriating crops to
centers of diversity, developing new U.S. crops, and encouraging international trading diplomacy through free exchange of plant germplasm. 4. What were the most significant accomplishments this past year? A. Single most significant. We did this research because Medicago truncatula is being used by the genomics research community as a model species for legumes. We planted out the entire collection of M. truncatula (319 accessions) in a replicated field trial at Prosser, WA,where a bulk DNA was isolated from 10 plants of 250 accessions and a second set of leaf tissue was collected from 10 different plants for all accessions for DNA extraction and analysis for SSR markers. We have collected data on 40 morphological and agronomic traits, as well as come close to completing an SSR analysis on the Medicago truncatula collection. Results from this research will provide basic information needed to support research on functional genomics in the model species, M. truncatula, and these data
will not only encourage the use of the collections, but help us develop collections that broadly represent biological diversity with a manageable number of accession. A. Other significant accomplishments. We did this research to ensure USDA germplasm collections of Medicago, Trifolium and Lotus continue to meet the needs of a broad array of users and contribute to the global conservation of these important genetic resources. The NTFLGRU in Prosser, WA, carried out state-of-the-art seed regeneration, computer documentation, and research on regeneration and acquisition methods in cooperation with Washington State University, Prosser, WA, and in collaboration with the Western Regional Plant Introduction Station, Pullman, WA. NTFLGRU produced seed on 400 accessions, including wild species and vulnerable seedlots, and distributed more than 2520 (from 10/01/02 to 6/30/03) seed packets. Collection management activities in 2003 will facilitate crop improvement among forage legume crops and
help in conserving valuable genetic resources. C. Significant activities. None D. None 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. This project was initiated in 01/29/03, so accomplishments to date are covered in Question 4. Replaces 5348-21000-015-00D which terminated 01/28/03. 6. What do you expect to accomplish, year by year, over the next 3 years? 2004: We plan to regenerate 400 accessions using routine field procedures and in vitro rescue techniques. We will collect basic characterization data on increased germplasm. Morphological characterization of M truncatula will be completed, as will molecular marker analysis. Data will be analyzed and core subset developed. We will also develop a collecting proposal that focuses on M. truncatula. Seed production protocols will be developed for Trifolium thompsonii. A seed- collecting trip will be made to sample the extant populations of Trifolium thompsonii. A base line
population inventory using GPS will also be made of Thompson clover. We will also collect other wild legumes if our FY03 proposal is funded. 2005: We plan to regenerate 400 accessions per year using routine field procedures and in vitro rescue techniques. We will collect basic characterization data on increased germplasm . We will make a collection of M. truncatula, if the FY04 proposal is funded. We will carry out molecular marker analysis on 1995 and 2004 origignal seed of Thompson clover. We will regenerate Thompson clover using our standard ex situ protocols. 2006: We expect to regenerate 400 accessions per year using routine field procedures and in vitro rescue techniques. This will include germplasm collected in 2005. We will collect basic characterization data on increased germplasm . We will carry out molecular marker analysis of increased populations of Thompson clover (provided we have been able to obtain seed off of 1-year-old plants. Data on 1994, 2004 and 2005
populations will be analyzed. 7. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? GPS and GIS technology was transferred to the Institute of Agricultural Genetics, in Hanoi, Vietnam in January 2003, in support of the the UNDP/GEF-VIE/01/G35 project entitled, "In situ conservation of Native Landraces and their wild relatives in Vietnam". This was done by teaching a short course on using GPS and GIS to support in situ conservation and providing technical support and advise for the project.
Impacts
(N/A)
Publications
- Bauchan, G.R., Greene, S.L. Status of the Medicago germplasm collection in the United States. Plant Genetic Resources Newsletter. 2002. v. 129. p. 1-8.
- Greene, S.L. Improving the quality of germplasm data to enhance germplasm use and management. Plant Genetic Resources Newsletter. 2001. v. 125. p. 1-8.
- Greene, S.L., Morris, J.B. The case for multiple-use plant germplasm collections and a strategy for implementation. 2001. Crop Science 41:886- 892.
- Morris, J.B., Greene, S.L. Defining a Multiple-use Germplasm Collection for the Genus Trifolium. 2001. Crop Science. v. 41. p. 893-901.
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