CONSERVATION, CHARACTERIZATION, AND EVALUATION OF CROP GENETIC RESOURCES AND ASSOCIATED INFORMATION

Goals / Objectives
Strategically expand the genetic diversity in genebank collections and improve associated information for priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources. Conserve and regenerate priority vegetable, sorghum, peanut, subtropical/tropical legume, new crop, and warm-season grass genetic resources efficiently and effectively, and distribute pathogen-tested samples and associated information worldwide. Strategically characterize (¿genotype¿) and evaluate (¿phenotype¿) priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources for molecular markers, morphological descriptors, and key agronomic or horticultural traits such as biochemical content and product quality. Conserve, regenerate, and distribute germplasm of specialty crops, current or potential bioenergy crops (e.g., sweet sorghum, switch grass, and Miscanthus), and new genetic stocks generated by genomic research (e.g. , assocition mapping projects) with sorghum and other crops.

Project Methods
Acquire samples of native warm-season grasses, bioenergy crops, subtropical legumes, Ipomoea species, chile pepper, and annual clovers to fill current gaps in NPGS collections. Survey existing holdings of sorghum genetic stocks, identify material that would fill gaps in NPGS collections, and begin acquiring and characterizing them. Conserve and distribute genomic research genetic stocks including association mapping populations of sorghum and other crops. Regenerate, conserve, and distribute more than 88,000 accessions of specialty crops, bioenergy crops, and other priority genetic resources and associated information. Increase the number of sweetpotato and warm-season grass clonal accessions maintained in tissue culture. Increase to 95 percent the proportion of the collection backed up at second sites. Develop superior regeneration methods for seed and clonally-propagated crops. Assay stored genetic resources for vigor, viability, and health. Distribute on request accessions and information that meet the specific needs of researchers and breeders. Develop and apply new genetic markers for phylogenetic and genetic diversity analyses of priority crops. Update and apply phenotypic descriptors for vegetables, peanuts, warm-season grasses, and subtropical/tropical legumes. Develop, enhance, and/or apply high performance liquid chromatography (HPLC) procedures for analyzing variation in flavonoids, antioxidants, capsaicin, and other key phytochemicals in accessions. Incorporate characterization, phenotypic, and biochemical data into GRIN and/or other databases.

Progress 04/22/08 to 03/18/13

Outputs
Progress Report Objectives (from AD-416): Strategically expand the genetic diversity in genebank collections and improve associated information for priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources. Conserve and regenerate priority vegetable, sorghum, peanut, subtropical/ tropical legume, new crop, and warm-season grass genetic resources efficiently and effectively, and distribute pathogen-tested samples and associated information worldwide. Strategically characterize (�genotype�) and evaluate (�phenotype�) priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources for molecular markers, morphological descriptors, and key agronomic or horticultural traits such as biochemical content and product quality. Conserve, regenerate, and distribute germplasm of specialty crops, current or potential bioenergy crops (e.g., sweet sorghum, switch grass, and Miscanthus), and new genetic stocks generated by genomic research (e. g., assocition mapping projects) with sorghum and other crops. Approach (from AD-416): Acquire samples of native warm-season grasses, bioenergy crops, subtropical legumes, Ipomoea species, chile pepper, and annual clovers to fill current gaps in NPGS collections. Survey existing holdings of sorghum genetic stocks, identify material that would fill gaps in NPGS collections, and begin acquiring and characterizing them. Conserve and distribute genomic research genetic stocks including association mapping populations of sorghum and other crops. Regenerate, conserve, and distribute more than 88,000 accessions of specialty crops, bioenergy crops, and other priority genetic resources and associated information. Increase the number of sweetpotato and warm-season grass clonal accessions maintained in tissue culture. Increase to 95 percent the proportion of the collection backed up at second sites. Develop superior regeneration methods for seed and clonally-propagated crops. Assay stored genetic resources for vigor, viability, and health. Distribute on request accessions and information that meet the specific needs of researchers and breeders. Develop and apply new genetic markers for phylogenetic and genetic diversity analyses of priority crops. Update and apply phenotypic descriptors for vegetables, peanuts, warm-season grasses, and subtropical/ tropical legumes. Develop, enhance, and/or apply high performance liquid chromatography (HPLC) procedures for analyzing variation in flavonoids, antioxidants, capsaicin, and other key phytochemicals in accessions. Incorporate characterization, phenotypic, and biochemical data into GRIN and/or other databases. This is the final report for Project Number 6607-21000-010-00D, entitled �Conservation, Characterization, and Evaluation of Crop Genetic Resources and Associated Information�; replaced by project 6607-21000-011-00D. During the five years of this project, the Griffin plant genetic resources collection increased from 88,270 accessions and 1,490 plant species to 92,180 accessions and 1,553 plant species. Availability improved slightly (87.3 to 87.9%) and security back up at a second location also improved (94.3 to 96.6%). Bulk seed samples of accessions maintained for long-term storage in -18 C improved (65% to 75%). Storing most seed in -18 C rather than 4 C improves long term viability and permits better retention of genetic variability for users. A total of 50, 930 accessions were distributed to users in 74 foreign countries and 121, 687 accessions were distributed to users in all 50 states. Four plant collection trips were conducted in the southern U.S. acquiring native switchgrass, naturalized Sorghum halepense, and other native warm-season grasses for the collection. Germination testing of accessions, first initiated at Griffin in 2002, improved from 51.4% of accessions tested to 84.4% of accessions tested. The number of images in GRIN of Griffin accessions increased from 9,800 to 18,600 and over 7.2 million data points were added to GRIN during this project. Seed regenerations and characterizations were conducted on over 6,300 peanut, cowpea, warm- season grass, pepper, watermelon, legume, new, and misc. crop, annual clover, and other vegetable accessions as well as several thousand sorghum accessions regenerated by cooperators in Puerto Rico and St. Croix. Long-term maintenance of several hundred wild peanut and warm- season grass clonal accessions was continued in the greenhouse. Several bermudagrass and over 720 sweetpotato accessions were maintained in tissue culture. A new method was developed to detect high oleic acid peanuts from seed or leaf tissue and was utilized to detect specific genotypes in peanut breeding. Genetic diversity was detected for flavonoids in Hibiscus species; capsaicin and capsiate in chile peppers; glucose and cellulosic traits in sunn hemp; salt tolerance and ploidy level in seashore paspalum; and salt tolerance in zoysia. The sweetpotato collection was screened for sweetpotato leaf curl virus and distribution of infected clones was restricted. Several or all accessions of the peanut mini-core, okra, sesame, castor bean, watermelon, lablab, pumpkin species, hibiscus, desmodium, and chile pepper collections were evaluated for oil content, fatty acid composition, and/or antioxidant compounds and data was added to GRIN. Alternate regeneration techniques were conducted on photoperiod-sensitive accessions using greenhouse, hydroponic, and aeroponic techniques. In cooperation with industry, differential sets of four vegetable crops were distributed to researchers for identification of disease races.

Impacts
(N/A)

Publications

Morris, J.B., Antonious, G. 2013. Glucose, stem dry weight variation, principal component and cluster analysis for some agronomic traits among 16 regenerated Crotalaria juncea accessions for potential cellulosic ethanol. Journal of Environmental Science and Health. 48(3):214-218.
Jarret, R.L., Levy, I.J., Potter, T.L., Cermak, S.C. 2013. Seed oil and fatty acid composition in Capsicum spp. Journal of Food Composition and Analysis. 30(2):102-108.
Morris, J.B., Wang, M.L., Tonnis, B.D. 2013. Variability of phenotype, anthocyanin indexes, and flavonoids in accessions from a close relative of soybean, Neontonia wightii (Wright & Arn. J.A. Lackey) in the U.S. germplasm collection for potential use as a health forage. In: El-Shemy, Hany A., editor.Soybean Bio-Active Compounds. Rijeka,Croatia:Intech. p.375- 386.
Jarret, R.L., Levy, I.J., Potter, T.L., Cermak, S.C., Merrick, L.C. 2013. Seed oil content and fatty acid composition in a genebank collection of Cucurbita moschata Duchesne and C. argyrosperma C. Huber. Plant Genetic Resources: Characterization and Utilization. 11:149-157.
Wang, J., Wang, M.L., Spiertz, J., Liu, Z., Han, L., Xie, G. 2013. Genetic variation in yield and chemical composition of wide range of sorghum accessions grown in north-west China. Research on Crops (an International Journal). 14(1):95-105.
Wang, M.L., Chen, C.Y., Tonnis, B.D., Barkley, N.L., Pinnow, D.L., Pittman, R.N., Davis, J., Holbrook Jr, C.C., Stalker, H., Pederson, G.A. 2013. Oil, fatty acid, flavonoid, and resveratrol content variability in FAD2A functional SNP genotypes in the U.S. peanut mini-core collection. Journal of Agricultural and Food Chemistry. 61:2875-2882.
Morris, J.B., Wang, M.L., Grusak, M.A., Tonnis, B.D. 2013. Fatty acid, flavonol, and mineral composition variability among seven macrotyloma uniflorum (Lam.) verdc. accessions. Agriculture 2013. 3(1):157-169.
Liu, J., Wang, M.L., Tonnis, B.D., Habteselassie, M., Liao, X., Huang, Q. 2012. Fungal pretreatment of switchgrass for improved saccharification and simultaneous enzyme production. Bioresource Technology. 135:39-45.
Morris, J.B., Grusak, M.A., Wang, M.L., Tonnis, B.D. 2013. Mineral, flavonoid, and fatty acid concentrations in ten diverse Lablab purpureus (L.) sweet accessions. In: Kuang, H., editor. Phytochemicals: Occurence in Nature, Health Effects and Antioxidant Properties. New York, NY, Nova Publishers. p. 219-224.
Xin, Z., Wang, M.L., Chopra, S., Wang, P. 2013. Gene Mutagensis Systems and Resources for the Saccharinae. In: Patterson, A.H., editor. Plant Genetics and Genomics: Crops and Models. New York, NY:Springer Publishing Company. 11:p. 169-185.
Morris, J.B. 2012. Showy partridge pea [Chamaecrista fasciculate (Michx.) Greene] with potential for cultivation as a multi-functional species in the United States. Genetic Resources and Crop Evolution. 59:1577-1581.
Barthe, S., Gugerli, F., Barkley, N.L., Maggia, L., Cardi, C., Scotti, I. 2012. Always look on both sides: Phylogenetic information conveyed by simple sequence repeat allele sequences. PLoS One. 7(7):e40699. doi:10. 1371/journal.pone.0040699.
Wang, M.L., Xin, Z., Tonnis, B.D., Farrell, G., Pinnow, D.L., Chen, Z., Davis, J., Yu, J., Hung, Y., Pederson, G.A. 2012. Evaluation of sweet sorghum as a feedstock by multiple harvests for sustainable bioenergy production. Journal of Sustainable Bioenergy Systems (JSBS). 2:122-137.

Progress 10/01/11 to 09/30/12

Outputs
Progress Report Objectives (from AD-416): Strategically expand the genetic diversity in genebank collections and improve associated information for priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources. Conserve and regenerate priority vegetable, sorghum, peanut, subtropical/tropical legume, new crop, and warm-season grass genetic resources efficiently and effectively, and distribute pathogen-tested samples and associated information worldwide. Strategically characterize (�genotype�) and evaluate (�phenotype�) priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources for molecular markers, morphological descriptors, and key agronomic or horticultural traits such as biochemical content and product quality. Conserve, regenerate, and distribute germplasm of specialty crops, current or potential bioenergy crops (e.g., sweet sorghum, switch grass, and Miscanthus), and new genetic stocks generated by genomic research (e.g., assocition mapping projects) with sorghum and other crops. Approach (from AD-416): Acquire samples of native warm-season grasses, bioenergy crops, subtropical legumes, Ipomoea species, chile pepper, and annual clovers to fill current gaps in NPGS collections. Survey existing holdings of sorghum genetic stocks, identify material that would fill gaps in NPGS collections, and begin acquiring and characterizing them. Conserve and distribute genomic research genetic stocks including association mapping populations of sorghum and other crops. Regenerate, conserve, and distribute more than 88,000 accessions of specialty crops, bioenergy crops, and other priority genetic resources and associated information. Increase the number of sweetpotato and warm-season grass clonal accessions maintained in tissue culture. Increase to 95 percent the proportion of the collection backed up at second sites. Develop superior regeneration methods for seed and clonally-propagated crops. Assay stored genetic resources for vigor, viability, and health. Distribute on request accessions and information that meet the specific needs of researchers and breeders. Develop and apply new genetic markers for phylogenetic and genetic diversity analyses of priority crops. Update and apply phenotypic descriptors for vegetables, peanuts, warm-season grasses, and subtropical/tropical legumes. Develop, enhance, and/or apply high performance liquid chromatography (HPLC) procedures for analyzing variation in flavonoids, antioxidants, capsaicin, and other key phytochemicals in accessions. Incorporate characterization, phenotypic, and biochemical data into GRIN and/or other databases. A total of 91,259 accessions of 1,548 plant species were maintained in the Griffin plant genetic resources collection. Over 87.8% of accessions were available for distribution to users and over 97.2% were backed up for security at a second location. Bulk seed samples for 67,241 accessions were maintained at -18 C for long-term storage with seed of the remaining accessions stored at 4 C. A total of 32,512 seed and clonal accessions in 946 separate orders were distributed upon request to scientists and educators in 47 U.S. states and 45 foreign countries. Acquisitions made to the collection included 176 sorghum, 230 pepper, 90 warm-season grass, 32 cowpea, 12 peanut, 13 vegetable, and 10 other accessions. A plant collection trip in Alabama, Florida, Georgia, and South Carolina added 39 naturalized Sorghum halepense, 14 switchgrass, and 3 indiangrass accessions to the collection. Seed regenerations and characterization were conducted on 815 peanut, 77 cowpea, 113 warm-season grass, 150 pepper, 301 legume, new, and misc. crop, 127 annual clover, and 32 other vegetable accessions. Peanut accessions were regenerated with cooperators in Georgia, Florida, North Carolina, Oklahoma, and New Mexico. Over 200 pepper accessions were grown in California for characterization and recording of digital images. Digital images of sorghum, cucurbit, cowpea and warm-season grass accessions, and seed oil and fatty acid content of okra and watermelon, peanut core fatty acid content, and sorghum 100-seed weight data were added to the Germplasm Resources Information Network (GRIN). Long-term maintenance of 242 wild peanut and 411 warm-season grass clonal accessions was continued in the greenhouse with an additional 47 napiergrass accessions maintained in the field. A total of 24 bermudagrass and 725 sweetpotato accessions were maintained in tissue culture. Germination testing has been conducted on 73,274 accessions (over 81% of collection) since 2002. In cooperation with industry, differential sets of four vegetable crops were distributed to researchers for identification of disease races. Photoperiod-sensitive Teramnus and annual clover accessions were regenerated in the winter greenhouse. Seed oil content and fatty acid composition was determined for the entire U.S. collections of kenaf, roselle, okra (oil content only) , and two pumpkin species. Morphological descriptor, oil content, fatty acid composition, and genetic variability data are being used to develop a core subset from the U.S. castor bean collection. With cooperators in Kansas, a total of 300 sorghum accessions were selected for biomass evaluation in the field from 1,000 accessions genotyped by genotyping-by- sequencing (GBS) analysis. Variation for phytochemicals including flavonol content, oil content, fatty acid composition, and anthocyanin index were determined in lablab, desmodium, roselle, and/or kenaf accessions. Variation in glucose content was found in sunn hemp accessions. Six clonal little bluestem lines were evaluated in the field for ornamental cultivar development. Salt tolerance screening was conducted on the U.S. zoysia collection. Significant Activities that Support Special Target Populations: Scientists mentored students at Griffin High School, a predominately minority school, giving advice on science fair projects and cleaned a nature trail at the school during the USDA Day of Service. High school and middle school students, including minority students, were mentored online on classroom germination experiments at schools in Illinois, Florida, and Ohio through the PlantingScience.org program. In conjunction with the University of Georgia Young Scholars program, ARS employees at Griffin mentored minority high school students during the summer to promote science, technology, engineering, and mathematics to students typically underrepresented in these career areas. Presentations on ARS seed conservation were made to diverse audiences during Farm City Days at Griffin and two Peachtree City Library programs at Peachtree City. Accomplishments 01 Improved Storage of Plant Genetic Resources. Most plant genetic resourc are maintained under short-term (4C) rather than long-term (-18C) storag temperatures, which increases the need for frequent seed regeneration th can reduce genetic variability. ARS researchers at Griffin, GA, split se samples of over 67,000 accessions (75% of the collection) with the bulk each accession placed in -18C long-term storage. New storage facilities will enable most seed of the entire collection at Griffin to be stored i long-term storage. These plant genetic resources will remain viable long with reduced need for regeneration and better retention of genetic variability of the original sample for users. 02 Oil and Fatty Acid Content of Two Pumpkin Species. Pumpkin species are widely cultivated for seed that produces a high quality oil, however, little information is available on species variability for seed oil content or composition. ARS researchers at Griffin, GA, determined that average oil content was similar in two pumpkin species, but one subspeci (Cucurbita argyrosperma subsp. argyrosperma var. callicarpa) had greater seed oil content than others evaluated. Linoleic acid was identified as the predominant fatty acid in all samples analyzed of the two pumpkin species. These accessions will be used by researchers interested in developing pumpkin cultivars as an oil seed crop. 03 High Oleic Acid Detection in Peanut. The high oleic acid trait in peanu is an important seed quality trait of great interest in peanut cultivar development. ARS researchers at Griffin, GA, compared three different methods used by researchers to detect the high oleic acid trait. The genotyping method and capillary electrophoresis method were the most compatible methods in detecting high oleic acid peanuts, while the near infrared method was not as effective. These data provide information on the accuracy of high oleic acid detection for breeders who employ different platforms for detection. 04 Oil and Fatty Acid Content of Nine Chili Pepper Species. Chili peppers are cultivated primarily for their fresh fruit; however, a market exists for the oil extracted from pepper seed. Little information is available variability among the various types and species of chili pepper for the amount of oil present in seed or the composition of the oil. ARS researchers at Griffin, GA, found that seed oil content varied greatly from 11 to 36% among 250 accessions of nine chili pepper species, with Capsicum annum having higher average seed oil content than other species Linoleic acid was the predominant fatty acid present in seed of all species, though differences in seed fatty acid composition among species were observed. These results will facilitate research exploring the potential of chili pepper as an oil seed crop. 05 Acquisition of Native Warm-Season Grass Accessions. Native warm-season grasses are currently of interest to users for habitat restoration, biofuel use, or ornamental use. ARS researchers in Griffin, GA, acquired 90 native switchgrass, deertongue, Florida paspalum, indiangrass, and se oats accessions for the U.S. warm-season grass collection. These grasses will provide users with greater genetic variaibilty needed for their breeding and research program. 06 Seed Oil and Fatty Acid Content of U.S. Collection of Hibiscus. A numbe of oil seed crops are under evaluation for potential biofuel or bioproducts use, however little information is available on the genetic variability for oil content among plant genetic resources of these crops ARS researchers at Griffin, GA, identified two accessions from 329 Hibiscus accessions with higher oil content of over 22%. Also, one Hibiscus accession was identified with much greater vernolic acid conten which may be used for industrial oil production. These accessions have potential for breeding Hibiscus with greater, high quality oil productio for use in developing new bioproducts.

Impacts
(N/A)

Publications

Barkley, N.L., Pinnow, D.L., Wang, M.L., Ling, K., Jarret, R.L. 2011. Detection and classification of SPLCV isolates in the U.S. sweetpotato germplasm collection via a real-time PCR assay and phylogenetic analysis. Plant Disease. 95(11):1385-1391.
Jenkins, T.M., Wang, M.L., Barkley, N.L. 2012. Microsatellite markers in plants and insects part II: Databases and in silico tools for microsatellite mining and analyzing population genetic stratification. Genes, Genomes, and Genomics. 6(1):60-75.
Wang, M.L., Chen, C.Y., Pinnow, D.L., Barkley, N.L., Pittman, R.N., Lamb, M.C., Pederson, G.A. 2012. Seed dormancy variability in the U.S. peanut mini-core collection. Research Journal of Seed Science. 5:84-95.
Wang, M.L., Raymer, P., Chinnan, M., Pittman, R.N. 2012. Screening of the US peanut germplasm for oil content and fatty acid composition. Biomass and Bioenergy. 39:336-343.
Jarret, R.L., Levy, I. 2012. Oil and fatty acid content in seed of Citrullus lanatus Schrad. Journal of Agricultural and Food Chemistry. 60(20):5199-5204.
Wu, Y., Li, X., Xiang, W., Zhu, C., Lin, Z., Wu, Y., Li, J., Pandravada, S. , Ridder, D., Bai, G., Wang, M.L., Trick, H., Bean, S., Tuinstra, M., Tesso, T., Yu, J. 2012. Presence of tannins in sorghum grains is conditioned by different natural allels of Tan1. Proceedings of the National Academy of Sciences. doi:10.1073/pnas.1201700109/-/DCSupplemental.
Wang, M.L., Morris, J.B., Tonnis, B.D., Davis, J., Pederson, G.A. 2012. Assessment of oil content and fatty acid composition variability in two economically important Hibiscus species. Journal of Agricultural and Food Chemistry. 60:6620-6626.
Fountain, J.C., Qin, H., Chen, C.Y., Dang, P.M., Wang, M.L., Guo, B. 2011. A note on development of a low-cost and high throughput SSR-based genotyping method in peanut (Arachis hypoghea L.). Peanut Science. 38:122- 127.
Lin, Z., Li, X., Shannon, L.M., Yeh, C., Wang, M.L., Bai, G., Peng, Z., Li, J., Trick, H.N., Clemente, T.E., Doebley, J., Schnable, P.S., Tuinstra, M. R., Tesso, T.T., White, F., Yu, J. 2012. Parallel domestication of the Shattering1 genes in cereals. Nature Genetics. 44:720-724.
Xin, Z., Wang, M.L. 2011. Sorghum as a versatile feedstock for bioenergy production. Biofuels. 2(5):577-588.
Morris, J.B., Hellier, B.C., Connett, J.F. 2011. Medicinal properties of legumes. In: Singh, R., editor. Genetic Resources, Chromosome Engineering, and Crop Improvement Medicinal Plants. Vol.6. Urbana,IL:CRC Press. p.297- 396.
Morris, J.B. 2011. Christmas-candle Senna:An ornamental and pharmaceutical plant. In: Singh R., editor. Genetic Resources, Chromosome Engineering, and Crop Improvement Medicinal Plants. Vol.6. Urbana,IL:CRC Press. p.793
Morris, J.B., Wang, M.L., Thomas, T. 2012. Quercetin, kaempferol, myricetin, and fatty acid content among several Hibiscus sabdariffa accession calyces based on maturity in a greenhouse. In: Chikamatsu, T., Hida, Y., editors. Quercetin: Dietary sources, functions and health benefits. Hauppauge,NY:Nova Science Publishers. p.269-282.
Mosjidis, J.A., Wang, M.L. 2011. Crotalaria. In: Chittaranjan, K.,editor. Wild Crop Relatives:Genomic and Breeding Resources Industrial Crops. 1st edition. New York,NY:Springer. p.63-69.

Progress 10/01/10 to 09/30/11

Outputs
Progress Report Objectives (from AD-416) Strategically expand the genetic diversity in genebank collections and improve associated information for priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources. Conserve and regenerate priority vegetable, sorghum, peanut, subtropical/tropical legume, new crop, and warm-season grass genetic resources efficiently and effectively, and distribute pathogen-tested samples and associated information worldwide. Strategically characterize (�genotype�) and evaluate (�phenotype�) priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources for molecular markers, morphological descriptors, and key agronomic or horticultural traits such as biochemical content and product quality. Conserve, regenerate, and distribute germplasm of specialty crops, current or potential bioenergy crops (e.g., sweet sorghum, switch grass, and Miscanthus), and new genetic stocks generated by genomic research (e.g., assocition mapping projects) with sorghum and other crops. Approach (from AD-416) Acquire samples of native warm-season grasses, bioenergy crops, subtropical legumes, Ipomoea species, chile pepper, and annual clovers to fill current gaps in NPGS collections. Survey existing holdings of sorghum genetic stocks, identify material that would fill gaps in NPGS collections, and begin acquiring and characterizing them. Conserve and distribute genomic research genetic stocks including association mapping populations of sorghum and other crops. Regenerate, conserve, and distribute more than 88,000 accessions of specialty crops, bioenergy crops, and other priority genetic resources and associated information. Increase the number of sweetpotato and warm-season grass clonal accessions maintained in tissue culture. Increase to 95 percent the proportion of the collection backed up at second sites. Develop superior regeneration methods for seed and clonally-propagated crops. Assay stored genetic resources for vigor, viability, and health. Distribute on request accessions and information that meet the specific needs of researchers and breeders. Develop and apply new genetic markers for phylogenetic and genetic diversity analyses of priority crops. Update and apply phenotypic descriptors for vegetables, peanuts, warm-season grasses, and subtropical/tropical legumes. Develop, enhance, and/or apply high performance liquid chromatography (HPLC) procedures for analyzing variation in flavonoids, antioxidants, capsaicin, and other key phytochemicals in accessions. Incorporate characterization, phenotypic, and biochemical data into GRIN and/or other databases. A total of 90,942 accessions of 1,534 plant species were maintained in the Griffin plant genetic resources collection. Over 87.7% of accessions were available for distribution to users and over 97.1% were backed up for security at a second location. Bulk seed samples for 66,995 accessions were maintained at -18 C for long-term storage with seed of the remaining accessions stored at 4 C. A total of 28,308 seed and clonal accessions in 925 separate orders were distributed upon request to scientists and educators in 49 U.S. states and 47 foreign countries. Acquisitions made to the collection included 143 sorghum, 144 warm-season grass, 19 vegetable and 2 other accessions. A plant collection trip was conducted in Alabama, Florida, Georgia, and South Carolina for naturalized Sorghum halepense germplasm that is in demand for sorghum gene flow studies. Seed regenerations and characterization were conducted on 299 peanut, 131 cowpea, 46 warm-season grass, 60 pepper, 361 legume, new, and misc. crop, 91 annual clover, and 30 other vegetable accessions. Over 300 pepper accessions were grown in the field for regeneration, characterization and recording of digital images. Digital images of cowpea, sorghum, cucurbit, and watermelon accessions, and pepper capsaicin content, peanut core selection, sorghum, and sweet sorghum data were added to the Germplasm Resources Information Network (GRIN). Long-term maintenance of 241 wild peanut and 458 warm-season grass accessions was continued in the greenhouse. Over 30 warm-season grass accessions and 712 sweetpotato accessions were maintained in tissue culture with eight replications of each sweetpotato clone. Germination testing has been completed for 69,556 accessions (over 77% of collection) since 2002. In cooperation with vegetable industry plant pathologists, differential sets of four vegetable crops are being established and distributed to researchers for identification of disease races. Fatty acid content was determined for the entire U.S. castor bean (over 1,000 accessions) and sesame (over 1,200 accessions) collections and 98 okra accessions. Over 660 castor accessions were genotyped with 15 Simple Sequence Repeats (SSR) markers for further genetic classification. A population of over 1,900 mutant watermelon seeds were developed for a mutant Targeting Induced Local Lesions in Genomes (TILLING) study with ARS cooperators in Charleston, SC. Ploidy level was determined for the entire St. Augustine and seashore paspalum grass collections. Photoperiod-sensitive Neonotonia, Teramnus, and annual clover accessions were successfully regenerated in the greenhouse. A total of 92 peanut accessions were successfully evaluated under quarantine and disease-free seed was produced. In association with Kansas State University scientists, 1,000 biomass sorghum accessions will be evaluated for plant morphology, biochemical composition, and genotype. Accomplishments 01 Fatty acid content of the U.S. sesame and castor bean collections. Plan accessions need to have not only high oil content, but proper fatty acid content to be utilized for biodiesel production. Oil content had previously been determined for the entire U.S. sesame and castor bean collections. ARS researchers at Griffin, GA, identified a sesame accession with both high oil and high oleic acid content, and a castor bean accession with high oil content and desirable fatty acid compositio These accessions will be useful for researchers developing crop cultiva suitable for biodiesel production from these plant species. 02 Phenotyping and genotyping peanut genetic resources. The high oleic tra in peanut is an important seed quality trait caused by two key mutations in the ahFAD2 gene. The nine genotypes that can be produced by crossing normal oleic to high oleic peanuts have never been characterized because there was not a method developed to track all possible genotypes and lin them to each phenotype. Therefore, a method was developed to detect all genotypes and the resulting phenotype (fatty acid composition) of each possible genotype was quantified in 500 individuals. This study demonstrated that the genetics of the trait were more complex than previously thought. Further, being able to detect all possible genotype can now expedite the breeding process by allowing unwanted characteristi to be purged from the population at early stages and only maintaining an evaluating the plants with the desired traits. This data provided information on the genetics controlling the trait, a method to detect al possible genotypes, and linked a phenotype with each respective genotype 03 Oil content in seed of watermelon species. Watermelon (Citrullus spp.) cultivated primarily in Africa for its use as an oil seed crop with oil having industrial and nutritional value. ARS researchers at Griffin, GA evaluated more than 1,000 accessions of various watermelon species for total oil content using nuclear magnetic resonance (NMR). Data indicate that total oil content varied from approximately 14% to >40%. Seed of egusi-type melons were notably higher in oil content than other types. This information will enable researchers to select watermelon accessions with potential for use as an oil seed crop. 04 Seashore paspalum salt tolerance and ploidy level. Plant breeders need know the number of chromosome sets or ploidy level of accessions in orde to produce fertile or sterile progeny. The ploidy level and salt tolerance of the entire U.S. seashore paspalum collection was determined The ploidy level and salt tolerance data will enable plant breeders to efficiently use the proper accessions within their breeding program. 05 Capsaicin and Capsiate in Capsicum species. The sensory attributes (chemical composition) of vegetables can be an important factor in determining consumer acceptance and demand. The genetic basis for the preferential synthesis of capsiate rather than capsaicin was investigate in a mutant line of chile pepper (Capsicum annuum). Capsiate is a non- pungent analog of capsaicin with documented medicinal properties. Data indicated that the mutation is inherited as a single recessive gene. Portions of the gene were sequenced and the mutate allele characterized. This information will be of benefit to those developing pepper lines or varieties enhanced for either of these compounds. 06 Alternate utilization of non-viable peanut seeds. The USDA maintains a germplasm collection of Arachis species which contains cultivated and wi peanuts. However, due to the high oil content in these seeds, the germination rate is known to drastically decline after 10-15 years or mo of cold storage. Low or zero viability (non-viable) seeds are often deemed useless in germplasm repositories. ARS scientists at Griffin, GA demonstrated that high quality Deoxyribo Nucleic Acid (DNA) can be obtained from non-viable seeds in wild and cultivated peanuts. The extracted DNA can be successfully used to reveal differences among individuals. This study shows that non-viable seed can be successfully used in molecular research.

Impacts
(N/A)

Publications

Gremillion, S.K., Culbreath, A.K., Gorbet, D.W., Mullinix Jr, B.G., Pittman, R.N., Stevenson, K.L., Todd, J.W., Escobar, R.E., Condori, M.M. 2011. Field evaluations of leaf spot resistance and yield in peanut genotypes in the United States and Bolivia. Plant Disease. 95(3):263-268.
Barkley, N.L., Wang, M.L., Pittman, R.N. 2011. A real-time PCR genotyping assay to detect FAD2A SNPs in peanuts (Arachis hypogaea L.). Electronic Journal of Biotechnology. 14(1). Available:
Jarret, R.L., Wang, M.L., Levy, I.J. 2011. Seed oil and Fatty acid content in okra (Abelmoschus esculentus) and related species. Journal of Agricultural and Food Chemistry. 59(8):4019-4024.
Chen, X., Wang, M.L., Holbrook Jr, C.C., Culbreath, A., Liang, X., Brennenman, T., Guo, B. 2010. Identification and characterization of a multigene family encoding germin-like proteins in cultivated peanut (Arachis hypogaea L.). Plant Molecular Biology Reporter. 29:389-403.
Antonious, G.F., Snyder, J.C., Burke, T., Jarret, R.L. 2010. Screening Capsicum chinense fruits for heavy metals bioaccumulation. Journal of Environmental Health. 45:562-571.
Wang, M.L., Morris, J.B., Pinnow, D.L., Davis, J., Raymer, P., Pederson, G. A. 2010. A survey of the castor oil content, seed weight and seed-coat colour on the United States Department of Agriculture germplasm collection. Plant Genetic Resources: Characterization and Utilization. 8:229-231.
Wang, M.L., Barkley, N.L., Chinnan, M., Stalker, T., Pittman, R.N. 2010. Oil content and fatty acid composition variability in wild peanut species. Plant Genetic Resources. 8:232-234.
Morris, J.B., Wang, M.L. 2011. Anthocyanin indexes, quercetin, kaempferol, and myricetin concentration in leaves and fruit of Abutilon theophrasti Medik. genetic resources. Plant Genetic Resources: Characterization and Utilization. 1:1-3.
Wang, M.L., Barkley, N.L., Chen, Z., Pittman, R.N. 2011. FAD2 Gene Mutations Significantly Alter Fatty Acid Profiles in Cultivated Peanuts (Arachis hypogaea). Biochemical Genetics. DOI: 10.1007/s10528-011-9447-3.
Wang, M.L., Sukumaran, S., Barkley, N.L., Chen, Z., Chen, C.Y., Guo, B., Pittman, R.N., Stalker, H., Holbrook Jr, C.C., Pederson, G.A., Yu, J. 2011. Population structure and marker-trait association analysis of the U.S. Peanut (Arachis hypogaea L.) mini-core collection. Journal of Theoretical and Applied Genetics. DOI: 10.1007/s00122-011-1668-7.
Morris, J.B., Wang, M.L., Morse, S.A. 2011. Ricinus. Wild Crop Relatives: Genomic and Breeding Resources. 3:251-260.
Morris, J.B., Wang, M.L. 2011. Evaluation for morphological, reproductive, anthocyanin index, and flavonol traits in ornamental and nutraceutical producing Hibiscus species. Ornamental Plants: Types, Cultivation and Nutrition. Hauppauge, NY: Nova Publishers. p.111-127.
Morris, J.B. 2011. Morphological, phenological, and peproductive trait analysis for the pasture species, Siratro [Macroptilium atropurpureum (DC.) Urb.]. Tropical Grasslands. 44:266-273.

Progress 10/01/09 to 09/30/10

Outputs
Progress Report Objectives (from AD-416) Strategically expand the genetic diversity in genebank collections and improve associated information for priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources. Conserve and regenerate priority vegetable, sorghum, peanut, subtropical/tropical legume, new crop, and warm-season grass genetic resources efficiently and effectively, and distribute pathogen-tested samples and associated information worldwide. Strategically characterize (�genotype�) and evaluate (�phenotype�) priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources for molecular markers, morphological descriptors, and key agronomic or horticultural traits such as biochemical content and product quality. Conserve, regenerate, and distribute germplasm of specialty crops, current or potential bioenergy crops (e.g., sweet sorghum, switch grass, and Miscanthus), and new genetic stocks generated by genomic research (e.g., assocition mapping projects) with sorghum and other crops. Approach (from AD-416) Acquire samples of native warm-season grasses, bioenergy crops, subtropical legumes, Ipomoea species, chile pepper, and annual clovers to fill current gaps in NPGS collections. Survey existing holdings of sorghum genetic stocks, identify material that would fill gaps in NPGS collections, and begin acquiring and characterizing them. Conserve and distribute genomic research genetic stocks including association mapping populations of sorghum and other crops. Regenerate, conserve, and distribute more than 88,000 accessions of specialty crops, bioenergy crops, and other priority genetic resources and associated information. Increase the number of sweetpotato and warm-season grass clonal accessions maintained in tissue culture. Increase to 95 percent the proportion of the collection backed up at second sites. Develop superior regeneration methods for seed and clonally-propagated crops. Assay stored genetic resources for vigor, viability, and health. Distribute on request accessions and information that meet the specific needs of researchers and breeders. Develop and apply new genetic markers for phylogenetic and genetic diversity analyses of priority crops. Update and apply phenotypic descriptors for vegetables, peanuts, warm-season grasses, and subtropical/tropical legumes. Develop, enhance, and/or apply high performance liquid chromatography (HPLC) procedures for analyzing variation in flavonoids, antioxidants, capsaicin, and other key phytochemicals in accessions. Incorporate characterization, phenotypic, and biochemical data into GRIN and/or other databases. A total of 90,668 accessions of 1,546 plant species were maintained in the Griffin plant genetic resources collection. Over 87.9% of accessions were available for distribution to users and over 96.3% were backed up for security at a second location. Bulk seed samples for 62,524 accessions were maintained at -18 C for long-term storage with seed of the remaining accessions stored at 4 C. A total of 40,449 seed and clonal accessions in 899 separate orders were distributed upon request to scientists and educators in 50 U.S. states and 38 foreign countries. Acquisitions made to the collection included 354 sorghum, 125 peanut, 70 annual clover, 68 warm-season grasses, and 22 vegetable accessions. A plant collection trip was conducted in northern Florida for native switchgrass germplasm. Seed regenerations and characterization were conducted on 275 peanut, 103 warm-season grass, 70 pepper, 346 legume, new, and misc. crop, 42 annual clover, and 8 cucurbit accessions. New regeneration techniques were developed for Hibiscus and Bituminaria species. Over 300 pepper accessions were grown in the field for characterization and recording of digital images. Digital images of cowpea, sorghum, pepper, and warm-season grass accessions, and pepper root rot and watermelon root knot nematode data were added to the Germplasm Resources Information Network (GRIN). Long-term maintenance of 543 wild peanut and 410 warm-season grass accessions was continued in the greenhouse. Over 30 warm-season grass accessions and 750 sweetpotato accessions were maintained in tissue culture with eight replications of each sweetpotato clone. Virus screening was completed on the sweetpotato collection for sweetpotato leaf curl virus. Germination testing has been completed for 60,207 accessions (over 67% of collection) since 2002. A genotyping technique was developed to detect high oleic acid peanuts and will be useful in evaluating segregating populations to identify progeny with the desirable high oleic acid trait. Molecular analysis and ploidy level determinations are underway for newly acquired switchgrass germplasm from Florida. Basic descriptors were collected on the entire U. S. bamboo collection. The oil content was determined for the entire U.S. castor bean (over 1,000 accessions) and okra (over 1,200 accessions) collections and the U.S. sesame collection is currently being evaluated for oil content. Fifty castor bean accessions are being grown in the field to verify oil content and determine relationships with fatty acid composition. Okra accessions varying in oil content will be evaluated for fatty acid composition. In collaboration with other ARS scientists in Dawson, GA, the peanut mini-core collection was grown in the field for morphological evaluation, is being biochemically analyzed in the lab, and is being genotyped with 84 genetic markers. In collaboration with Kansas State University scientists, association analysis will be conducted to identify association of genetic markers with useful peanut traits. Accomplishments 01 Oil Content in Seed of Okra species. Okra (Abelmoschus spp.) is an oil seed crop that is known to produce oil yields with chemical properties similar to those of cotton seed. ARS researchers at Griffin, GA evaluate more than 1,200 accessions of various okra species for total oil content using nuclear magnetic resonance (NMR). Data indicated that total oil content varied from approximately 4% to >21% and that it was loosely correlated with seed weight. This information will enable researchers to select okra accessions with potential for use as an oil seed crop. 02 Oil Content of the U.S. Castor Bean Collection. Oil content for biodies production among the castor bean accessions in the U.S. collection is unknown. ARS researchers at Griffin, GA, found significant variation in oil content (ranging from 37 to 61%) in 1,033 castor bean accessions. Selected castor bean accessions with high oil content can now be used in variety development for use as a biodiesel crop. 03 Improvement of the Switchgrass Germplasm Collection. Evaluation of switchgrass accessions is needed in order to provide well characterized, available and viable material to scientists for use in bioenergy feedsto development. Newly collected switchgrass accessions were tested for germination, regenerated, characterized genotypically using simple sequence repeat (SSR) markers, and ploidy levels determined by ARS researchers at Griffin, GA. This effort will provide well characterized lowland switchgrass germplasm to plant breeders for incorporation into bioenergy feedstock development programs furthering the endeavor towards S. energy independence. 04 Capsaicin and Capsiate in Chile Pepper species. The sensory attributes (chemical composition) of vegetables can be an important factor in determining consumer acceptance and demand. ARS researchers at Griffin, investigated the occurrence and concentration of capsaicin and capsiate in Capsicum spp. Capsaicin confers the pungency (heat) associated with hot peppers and capsiate is a related compound. The levels of capsaicin and capsiate ranged from 0 (not present) to >1500 ug and >600 ug/g FW, respectively. Capsiate was present in most of the tested chile pepper accessions. This information will be of benefit to those developing pepp lines or varieties enhanced for either of these compounds. 05 Genotyping Assay Developed for High Oleic Acid Peanuts. ARS researchers in Griffin, GA, developed a genotyping assay using a real-time polymeras chain reaction to detect alleles relating to the high oleic acid trait i peanuts. This assay enables breeders to test seed or leaf tissue in initial crosses without using chemical analysis of ground up seeds. Development of this rapid assay will help identify key genotypes linked important agronomic traits, improve breeding efficiency by eliminating undesirable plants, and expedite the process of developing improved pean cultivars by decreasing the time and effort to characterize all of the generated progeny. 06 Techniques Developed to Regenerate Legume Species. Optimum regeneration techniques are unknown for several legume species. ARS researchers at Griffin, GA developed improved regeneration techniques for two legume species. Twelve photoperiod sensitive Hibiscus sabdariffa accessions wer successfully regenerated in a greenhouse during the offseason (November June). Six perennial Bituminaria bituminosa accessions overwintered and produced quality fruits and seed during the second year of growth. Quali seed of these legumes are now available for scientists to use in their research projects. 07 Fatty Acid Variability in Lablab purpureus. Little information is available regarding fatty acid variability in the U.S. Lablab purpureus collection. Significant variation for several fatty acids was found in seven Lablab purpureus accessions by ARS researchers in Griffin, GA. The accessions can now be used by breeders and scientists in the development of new products containing these fatty acids. 08 Biofuel Potential of Sunn Hemp Accessions. Cellulosic ethanol is expect to play a large role in biofuels, however cellulose and biomass variatio is unknown among the U.S. sunn hemp (Crotalaria juncea) collection. Glucose is a primary constituent in cellulose and was found to vary amon these 16 sunn hemp accessions. ARS researchers at Griffin, GA, also foun significant variation accessions for biomass, apical dominance, branchin open flowers, plant height, earliness, and seed production. Several sunn hemp accessions were identified as parental species for use in breeding programs for cultivar development.

Impacts
(N/A)

Publications

Mckinney, J.T., Nay, L.M., Dekoeyer, D., Reed, G.H., Wall, M., Palais, R., Jarret, R.L., Wittner, C. 2010. Mutation Scanning and Genotyping in Plants by High Resolution DNA Melting. Handbook of Plant Mutation Screening. pp 149-163.
Chen, Z., Wang, M.L., Barkley, N.L., Pittman, R.N. 2010. A Simple Allele- Specific PCR Assay for Detecting FAD2 Alleles in Both A and B Genomes of the Cultivated Peanut for High Oleate Trait Selection. Plant Molecular Biology Reporter. 28:542-548.
Barkley, N.L., Chamberlin, K.D., Wang, M.L., Pittman, R.N. 2009. Development of a real-time PCR genotyping assay to identify high oleic acid peanuts (Arachis hypogaea L.). Molecular Breeding. DOI 10. 1007/s11032-009-9338-z 25(3):541-548.
Morris, J.B. 2009. Morphological and reproductive characterization in Hyacinth bean, Lablab purpureus (L) Sweet germplasm with clinically proven nutraceutical and pharmaceutical traits for use as a medicinal food. Journal of Dietary Supplement. DOI: 10.1080/19390210909070830 6(3):263- 279
Wang, M.L., Chen, C.Y., Davis, J., Guo, B., Stalker, T., Pittman, R.N. 2009. Assessment of Oil content and fatty acid composition variability in different peanut subspecies and botanical varieties. Plant Genetic Resources. doi: 10.1017/S1479262109990177. 8:71-73
Morris, J.B. 2010. Morphological and reproductive characterization of guar genetic resources regenerated in Georgia, USA. Genetic Resources and Crop Evolution. 10.1007/s10722-010-9538-8.
Auld, D.L., Zanotto, M.D., Mckeon, T.A., Morris, J.B. 2009. Castor. Handbook of Plant Breeding. 4:317-332
Wang, M., Barkley, N.A., Jenkins, T.M. 2009. Microsatellite Markers in Plants and Insects Part I:Applications of Biotechnology. Genes, Genomes, and Genomics. 3(1):54-67.
Wang, M.L., Zhu, C., Barkley, N.L., Chen, Z., Erpelding, J.E., Murray, S., Tesso, T., Pederson, G.A., Yu, J. 2009. Assessment of Genetic Diversity and Population Structure of Accessions in the US Sweet Sorthum Germplasm Collection. Theoretical and Applied Genetics. Online DOI 10.1007/S00122- 009-1155-6 120:13-23.

Progress 10/01/08 to 09/30/09

Outputs
Progress Report Objectives (from AD-416) Strategically expand the genetic diversity in genebank collections and improve associated information for priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources. Conserve and regenerate priority vegetable, sorghum, peanut, subtropical/tropical legume, new crop, and warm-season grass genetic resources efficiently and effectively, and distribute pathogen-tested samples and associated information worldwide. Strategically characterize (�genotype�) and evaluate (�phenotype�) priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources for molecular markers, morphological descriptors, and key agronomic or horticultural traits such as biochemical content and product quality. Conserve, regenerate, and distribute germplasm of specialty crops, current or potential bioenergy crops (e.g., sweet sorghum, switch grass, and Miscanthus), and new genetic stocks generated by genomic research (e.g., assocition mapping projects) with sorghum and other crops. Approach (from AD-416) Acquire samples of native warm-season grasses, subtropical legumes, Ipomoea species, chile pepper, and annual clovers to fill current gaps in NPGS collections. Survey existing holdings of sorghum genetic stocks, identify material that would fill gaps in NPGS collections, and begin acquiring and characterizing them. Regenerate and conserve more than 86, 000 accessions of priority genetic resources and associated information. Increase the number of sweetpotato and warm-season grass clonal accessions maintained in tissue culture. Increase to 95 percent the proportion of the collection backed up at second sites. Develop superior regeneration methods for seed and clonally-propagated crops. Assay stored genetic resources for vigor, viability, and health. Distribute on request accessions and information that meet the specific needs of researchers and breeders. Develop and apply new genetic markers for phylogenetic and genetic diversity analyses of priority crops. Update and apply phenotypic descriptors for vegetables, peanuts, warm-season grasses, and subtropical/tropical legumes. Develop, enhance, and/or apply high performance liquid chromatography (HPLC) procedures for analyzing variation in flavonoids, antioxidants, capsaicin, and other key phytochemicals in accessions. Incorporate characterization, phenotypic, and biochemical data into GRIN and/or other databases. Significant Activities that Support Special Target Populations A total of 89,451 accessions of 1,529 plant species were maintained in the Griffin plant genetic resources collection. Over 87% of accessions were available for distribution to users and over 96% were backed up for security at a second location. Bulk seed samples for 60,269 accessions were maintained at -18 C for long-term storage with seed of the remaining accessions stored at 4 C. A total of 30,883 seed and clonal accessions in 881 separate orders were distributed upon request to scientists and educators in 47 U.S. states and 45 foreign countries. Acquisitions made to the collection included 94 switchgrass, 230 pearl millet, 67 finger millet, 31 Hibiscus, and 30 vegetable accessions. Seed regenerations and characterization were conducted on 587 peanut, 150 warm-season grass, 323 legume, new, and misc. crop, 106 annual clover, and 10 cucurbit accessions. New regeneration techniques were developed for two Hibiscus, several cucurbits, and one sweetpotato species. Over 600 pepper accessions were grown in the field for characterization and recording of digital images. Digital images of watermelon seed and characterization data for big bluestem were added to the Germplasm Resources Information Network (GRIN). A new tablet PC was used to facilitate collection of peanut descriptor data. Long-term maintenance of 525 wild peanut and 398 warm-season grass accessions was continued in the greenhouse. Over 30 warm-season grass accessions and 750 sweetpotato accessions were maintained in tissue culture with eight replications of each sweetpotato clone. Virus screening was conducted on the warm-season grass clonal collection (for maize dwarf mosaic and johnsongrass mosaic viruses) and sweetpotato collection (for sweetpotato leaf curl virus). Tomato spotted wilt virus was found on peanut and clover species in the greenhouse and accessions were evaluated for virus infection by field, lab, and molecular techniques. Germination testing has been completed for 54,054 accessions (over 60% of collection) since 2002. A genotyping technique was developed to detect high oleic acid peanuts and will be useful in evaluating segregating populations to identify progeny with the desirable high oleic acid trait. In collaboration with other ARS scientists, 800 sorghum mutant lines were evaluated by tilling by ten candidate genes and gene function for brown midrib mutants was identified. In collaboration with university and ARS scientists, 96 sweet sorghum accessions were genotyped with 95 markers to determine their genetic diversity and population structure. These accessions will be used as a panel for association mapping in sweet sorghum. The oil content and fatty acid composition was determined for 48 castor bean and 200 peanut accessions. To determine nutritional value of peanut accessions, over 130 peanut accessions or breeding lines were evaluated for resveratrol and isoflavonoid content, and ratio of oleic/linoelic acid. Genetic variability for mineral, flavonoid, and anthocyanin index were determined for lablab, roselle, and perennial soybean accessions. Genetic relatedness was determined among species of pepper and among species of sweetpotato.

Impacts
(N/A)

Publications

Antonious, G., Berke, T., Jarret, R.L. Pungency in Capsicum Chinense: Variation Among Countries of Origin. 2009 Journal of Environmental Science and Health. 44(B)(2): 179-184.
Morris, J.B., Pederson, G.A., Quesenberry, K., Wang, M.L. 2009. Clover. In R.J.Singh (ed). Genetic Resources, Chromosome Engineering, and Crop Improvement: Forage Crops. p.207-228.
Naeem, M., Khan, M., Morris, J.B. 2009. Agrobotanical attributes, nitrogen- fixation, enzyme activities and nutraceuticals and tyrosinase enzyme of hyacinth bean (Lablab purpureus L.) - a bio-functional medicinal legume.. American Journal of Plant Physiology. 4:58-69.
Jarret, R.L. 2008. DNA Barcoding in a Crop Genebank: Resolving the Capsicum annuum Species Complex.. The Open Biology Journal. 1:35-42.
Wang, M.L., Pinnow, D.L., Barkley, N.L., Pittman, R.N. 2009. Plant Resistance to TSWV and Seed Accumulation of Resveratrol within Peanut Germplasm and Its Wild Relatives in the US Collection. Plant Pathology Journal ISSN 1812-5387 . 8(2):53-61.
Xin, Z., Wang, M.L., Burow, G.B., Burke, J.J. 2009. An induced sorghum mutant population suitable for bioenergy research. BioEnergy Research. 2(1- 2):10-16.
Zhenbang, C., Wang, M.L., Waltz, C., Raymer, P. 2009. Genetic Diversity of Warm-Season Turfgrass: Seashore Paspalum, Bermudagrass, and Zoysiagrass Revealed by AFLPs. FLORICULTURE, ORNAMENTAL AND PLANT BIOTECHNOLOGY. Online ISSN 1749-0308 3:1 p.20-24.
Weimer, P.J., Morris, J.B. 2009. Grasses and Legumes for Bio-Based Products. In: Wedin, W.F., Fales, S.L. editors. Grassland: Quietness and Strength for a New American Agriculture. Madison, WI: American Society for Agronomy/Crop Science Society of America/Soil Science Society of America. p. 221-233.
Wang, M.L., Barkley, N.L., Gillaspie, A.G., Pederson, G.A. 2008. Phylogenetic relationships and genetic diversity of the USDA Vigna germplasm collection revealed by gene-derived markers and sequencing.. Genetical Research. 90(6):467-480.
Barkley, N.L., Pinnow, D.L., Wang, M.L., Pederson, G.A. 2009. First Report of Tomato Spotted Wilt Virus Infecting African Clover (Trifolium tembense) in Georgia. Plant Disease. 93(2):202.
Singh, S., Jarret, R.L., Russo, V.M., Shimkus, J.M., Majetich, G., Bushway, R., Perkins, L.B. 2009. Determination of capsinoids by HPLC-DAD in Capsicum ssp. Part I. Journal of Agricultural and Food Chemistry. DOI.10. 1021/jf8040287
Jarret, R.L., Berke, T., Baldwin, E.A., Antonious, G. 2009. Variability for Free Sugars and Organic Acids in Capsicum Chinense Jacq.. Chemistry and Biodiversity. 6(2):138-145.
Morris, J.B. 2009. Characterization of sesame (Sesamum indicum L.) germplasm regenerated in Georgia, U.S.A.. Genetic Resources and Crop Evolution. DOI 10.1007/S10722-009-9411-9.
Morris, J.B. 2008. Characterization of regenerated butterfly pea (Clitoria ternatea L.) accessions for morphological, phenology, reproductive and potential nutraceutical, pharmaceutical trait utilization.. Genetic Resources and Crop Evolution. 56:421-427

Progress 10/01/07 to 09/30/08

Outputs
Progress Report Objectives (from AD-416) Strategically expand the genetic diversity in genebank collections and improve associated information for priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources. Conserve and regenerate priority vegetable, sorghum, peanut, subtropical/tropical legume, new crop, and warm-season grass genetic resources efficiently and effectively, and distribute pathogen-tested samples and associated information worldwide. Strategically characterize (�genotype�) and evaluate (�phenotype�) priority vegetable, sorghum, peanut, subtropical/tropical legume, and warm-season grass genetic resources for molecular markers, morphological descriptors, and key agronomic or horticultural traits such as biochemical content and product quality. Approach (from AD-416) Acquire samples of native warm-season grasses, subtropical legumes, Ipomoea species, chile pepper, and annual clovers to fill current gaps in NPGS collections. Survey existing holdings of sorghum genetic stocks, identify material that would fill gaps in NPGS collections, and begin acquiring and characterizing them. Regenerate and conserve more than 86, 000 accessions of priority genetic resources and associated information. Increase the number of sweetpotato and warm-season grass clonal accessions maintained in tissue culture. Increase to 95 percent the proportion of the collection backed up at second sites. Develop superior regeneration methods for seed and clonally-propagated crops. Assay stored genetic resources for vigor, viability, and health. Distribute on request accessions and information that meet the specific needs of researchers and breeders. Develop and apply new genetic markers for phylogenetic and genetic diversity analyses of priority crops. Update and apply phenotypic descriptors for vegetables, peanuts, warm-season grasses, and subtropical/tropical legumes. Develop, enhance, and/or apply high performance liquid chromatography (HPLC) procedures for analyzing variation in flavonoids, antioxidants, capsaicin, and other key phytochemicals in accessions. Incorporate characterization, phenotypic, and biochemical data into GRIN and/or other databases. Significant Activities that Support Special Target Populations This project replaced 6607-21000-009-00D in late April 2008. Progress for most of Fiscal Year 2008 is reported in the annual report for project 6607-21000-009-00D and only progress since late April 2008 will be reported for this project. No milestones in this project were developed for any period shorter than 12 months, so progress for the first milestones of this project will be reported in the next annual report. Progress since late April 2008 relating to National Program 301 Plant Genetic Resources, Genomics, and Genetics Improvement, Component 1 Plant and Microbial Genetic Resource Management, Problem Statement 1A Efficiently and Effectively Manage Plant and Microbial Genetic Resources. Since April 2008, a total of 13,308 seed, tissue culture, and clonal accessions were distributed in 454 orders to researchers and educators throughout the United States and world. Curators conducted regeneration of accessions that were unavailable, had low viability, or were in demand by users. Regenerations of subtropical/tropical legume, peanut, pepper and other vegetable, annual clover, cowpea, and warm-season grass accessions are currently underway. Germination tests continue to be conducted on accessions with tests completed on 48,852 accessions since 2002. Progress since late April 2008 relating to National Program 301 Plant Genetic Resources, Genomics, and Genetics Improvement, Component 1 Plant and Microbial Genetic Resource Management, Problem Statement 1B Assess the Systematic Relationships and Genetic Diversity of Crop Genetic Resources. Phenotypic characterization was conducted on all accessions that were regenerated. Additional characterization included 120 pepper and 236 warm-season grass accessions for phenotypic descriptors, 200 pepper accessions for capsaicin and dihydrocapsaicin content, 50 peanut accessions for oil and fatty acid composition, 21 legumes for anthocyanin index, 7 Hibiscus spp. for flavonol composition, and 48 castor bean accessions for oil content. Sweet sorghum accessions have been genotyped with 50 DNA markers and use of additional markers is underway. Sorghum mutant lines obtained from cooperators have been tilled by ten candidate genes to trace genotype mutation to phenotype. Protocols have been developed to detect sweetpotato leaf curl virus using real-time polymerase chain reaction (PCR) and DNA extraction for evaluation of the entire sweetpotato collection for detection of this virus is in progress. Tomato spotted wilt virus was detected in accessions of an African clover, Trifolium tembense, in the greenhouse. Study of the relationship of leaf spot infection with resveratrol content in peanut accessions has been attempted, but leaf spot inoculation of seedlings was unsuccessful.

Impacts
(N/A)

Publications

Dida, M., Wanyera, N., Harrison Dunn, M.L., Bennetzen, J., Devos, K. 2008. Population Structure and Diversity in Finger Millet (Eleusine coracana) Germplasm.. Tropical Plant Biology 1: 131-141.
Wang, M.L., Pittman, R.N. 2008. Resveratrol Content in Seeds of Peanut Germlasm Quantified by HPLC.. Plant Genetic Resources: Characterization and Utilization. 7(1) 80-83 (2008).