Duration: 10/01/2009 to 09/30/2014

Administrative Advisor(s):

NIFA Reps:

Non-Technical Summary

Statement of Issues and Justification

Plant genetic resources (germplasm) are vital for safeguarding the future of U.S. and global agriculture. Germplasm is the raw material that underpins the maintenance and development of crops for food, feed, and fiber production. Always needed for traditional plant breeding, these resources are rapidly becoming increasingly important for biofuel production, plant biotechnology, and for the development of industrial products. The USDA-ARS, National Plant Germplasm System (NPGS) is responsible for maintaining diverse germplasm collections in the U.S. for researchers and breeders. A major NPGS germplasm repository is the Western Regional Plant Introduction Station (WRPIS) at Washington State University, Pullman, Washington, which has long been part of the W-6 Regional Research Project. This project, a cooperative endeavor between the USDA-ARS and the Western State Agricultural Experiment Stations (SAESs), provides access to a wide range of plant genetic resources for scientists and stakeholders in the U.S., with emphasis on scientists and stakeholders in the 13 western states and land-grant universities. The W-6 Project (Plant Genetic Resource Management, Preservation, Characterization and Utilization) funds one of four Plant Introduction (PI) Stations in the NPGS, and like the other three PI projects, the W-6 project focuses on germplasm acquisition, conservation, regeneration, evaluation, documentation, and distribution. The renewal of this project is critical to supplying the genetic resources needed to maintain and develop food and fiber crops important to the Western Region and the entire U.S. The project will extend the availability of native species needed for revegetation, enhance the application of molecular marker technologies in key collections such as lettuce, and support disease and pest management in threatened collections such as garlic and chickpea.

Most agricultural crops are not native to the U.S. Thus, a continued supply of new, exotic germplasm is needed for ongoing genetic improvement of crops to support U.S. agriculture. Germplasm is required to provide the necessary genetic traits for U.S. food security, agricultural profitability, and competiveness (Duncan, 1995; Qualset and Shands, 2005; Day-Rubenstein and Heisey, 2006). The WRPIS holds approximately 80,000 germplasm accessions representing diverse crop species and their wild relatives. These species can be roughly divided into ten groups: 1) forage and turf grasses, 2) cool season food legumes (pea, lentil, chickpea, fava bean, lupine, etc.), 3) forage legume crops, 4) beans, 5) lettuce, 6) safflower, 7) garlic and onion relatives, 8) sugar beet, 9) selected ornamentals, and 10) medicinal plant species. The WRPIS holdings account for 15.5% of 509,000 accessions in the NPGS, which comprises 25 seed and clonal repositories including the related Regional Research Projects (North Central [NC-7] at Ames, IA; Northeastern [NE-9] at Geneva, NY; and Southern [S-9] at Griffin, GA). Most WRPIS accessions are maintained as seed, with a small proportion (garlic and relatives and some ornamentals) vegetatively-propagated.

Scientists affiliated with western U.S. Agricultural Experiment Stations (SAESs) and land-grant universities conduct research and extension projects that involve most of the crop species at the WRPIS. For example, forage and turf grass accessions are utilized for improving pasture and turf and for restoration of public and private lands in the western U.S. Lettuce is the leading vegetable crop in the U.S. with an annual value totaling more than $2 billion in 2004, and two western states, California and Arizona, account for more than 90 percent of U.S. lettuce production. Cool season food legumes (chickpea, pea, and lentil) are major crops in North Dakota, Washington and Idaho, with additional substantial acreage in California, Montana and elsewhere. In addition to the Western States, the W6 project fills germplasm needs nationwide, such as the lentil industry in North Dakota, beans in Michigan, and forage and turf grasses throughout the Midwest and Atlantic states. The continuation of the W-6 project will ensure that needed crop genetic resources are maintained as high quality, healthy and viable plant germplasm for researchers and breeders in the western U.S. and elsewhere.

Scientists and curators at the WRPIS utilize current methods and information to accomplish the major functions of acquiring, preserving, regenerating, evaluating, documenting and distributing germplasm accessions In addition, the two CRIS projects at the WRPIS (Plant Germplasm Introduction and Testing Research Unit, Pullman, Washington; National Temperate Forage Legume Genetic Resources Unit, Prosser, Washington) provide locations for re-generating germplasm under vastly different environmental conditions to serve the needs of crop species maintained by the WRPIS. Moreover, accomplishing this work requires that WRPIS scientists and curators cooperate with faculty and extension personnel in agronomy, horticulture, plant pathology, entomology, genetics, and plant physiology at Washington State University (WSU) and other western land-grant universities. For example, the WRPIS entomologist cooperates with scientists in the Plant, Soil and Entomological Sciences at the University of Idaho in evaluating cool season food legume germplasm for insect resistance, and the WRPIS agronomist collaborates with scientists of BLM (Bureau of Land Management) and in identifying suitable genotypes of native plant species for public land revegetation in the Great Basin. The WRPIS plant pathologist cooperates with WSU and other ARS scientists on biological and chemical control of Ascochyta blight of chickpea, and on identification and taxonomy of powdery mildew species on cool season legumes. Similarly, the WRPIS agronomy curator has a long history of cooperating with stakeholders at several forage grass seed companies in Oregon and elsewhere.

Collaborative projects also exist with scientists at international centers (e.g., ICRISAT- India; Biodiversity International, Rome, Italy; Centraalbureau voor Schimmelcultures, Utrecht, Netherlands), foreign institutes (John Innes, England), foreign national programs and foreign companies (AgResearch, New Zealand). The WRPIS Cool Season Food Legumes curator, working with scientists in ICARDA has acquired 1368 pea accessions of the Vavilov Institute Pisum collection. WRPIS accession holdings have increased at a rate of approximately 2,000 per year over the past five years (Attachment Figure 1: Growth in WRPIS Germplasm Collections). These collaborative activities highlight the importance of WRPIS in promoting the acquisition and utilization of new and existing germplasm.

The plant researchers in both public and private sectors frequently request germplasm from NPGS. Over the past seven years scientists from the Western Region, requested and received a total of 266,627 packets of seed samples from NPGS, and 46,843 packets were distributed from WRPIS. Most of these requests were from scientist at SAES Universities. Each year WRPIS sent out more than 20,000 seed packets to requesters worldwide, approximately 64% went to US users and 36% went to foreign countries. This germplasm is highly utilized. For example, over a five year period, 11% of germplasm distributed internationally had already been incorporated into breeding programs. This is well above the success rate typical for most "bioprospecting" endeavors.

Seed stocks at the WRPIS that are low in viability or supply must be regenerated. Advice from Crop Germplasm Committees, stakeholders, as well as the knowledge and experience of curators guide decisions concerning the timing and methodology of regenerations for specific seed collections. For this activity, priority is given to accessions that have not been regenerated, those with few seeds and/or low seed germination, and those that have not been duplicated at a back-up site (Clark et al., 1997; Jarret, 2006). Low quantity and quality seed arriving at the WRPIS and other genebanks must be regenerated before distributions to stakeholders can occur.

The biggest challenge for a regeneration program is to maintain the genetic integrity of the original collection. The first requirement for high quality regeneration is to understand the breeding systems of diverse taxa and design appropriate protocols that safeguard against physical and genetic mixing of accessions and natural selection and genetic drift (Frankel et al., 1995). From the limited literature on specific germplasm conservation (Pincker et al., 1987, Roath 1989, Rowe 1986), some general rules can be deduced and applied. In this context, WRPIS scientists and curators have developed seed-regeneration programs to safeguard the genetic integrity of specific collections. For insect pollinated taxa, the choices for insect pollinators and controlled pollination systems are critical (Clement et al., 2006; Clement et al., 2007a, 2007d). Indeed, there is a requirement for continued research on the pollination systems of many of the 2700 taxa in WRPIS collections because these taxa exhibit a wide diversity of breeding systems in the plant kingdom. Pollen isolation, effective population size, genetic drift, and unintended selection are important considerations for self-incompatible, outcrossing, entomophilous, and wind pollinated accessions (Clement et al., 2007; Johnson et al., 1996; Johnson et al., 2004). Based on genetic and statistical analyses, Marshall and Brown (1975) proposed a theoretical population size (sample size stored in collections and for sample size distributed) that would conserve at least one copy of each allele occurring at a frequency of >0.05 at the 0.95 probability level. However, limited budgets and human resources, as well as low numbers of seeds, can lead to low numbers of plants for some accessions in regeneration nurseries. This practice could potentially result in a loss of alleles that exist at very low frequencies. Also, there are few published guidelines on the regeneration of the wild relatives of several crop species (Kaiser et al., 1997 Muehlbauer et al., 1994), and none for many of the "new crop" species. For example, Russian dandelion germplasm is needed in breeding programs to fulfill a Federal mandate to develop a domestic source of natural rubber. The WRPIS horticulture curator collected from seed and root samples from Kazakhstan in 2008. However, no information on the reproductive biology is available for this species. These 'regeneration challenges' illustrate the requirement for continuous development of new regeneration protocols for the many germplasm holdings in the WRPIS (Johnson et al., 2004).

Biotic stresses (i.e. insect and disease) that adversely affect WRPIS germplasm maintenance efforts must be controlled to ensure the production of high quality seed for distribution to stakeholders. At WRPIS, research to assess etiologies of diseases and positive or negative interactions between arthropods and host plants have been an important part of the W-6 evaluation program for many years (Clement et al., 1990, 1991, 1993, 1997, 1999, 2002, 2005, 2007, Coyne et al., 2008, Dugan 2007, Dugan and Peever 2002, Dugan and Glawe 2006, 2007, Dugan and Newcombe 2007, Dugan et al., 2003, 2005, 2007, 2008). This type of research will continue as new problems develop, exemplified by recently discovered disease infections and mite infestations involving the WRPIS garlic collection. Additionally, knowledge of the relationships between insect vectors and plant pathogens (viral, bacterial, fungal), and plant- beneficial microbe associations (e.g., Clement et al., 2008), is essential for optimal seed regeneration and preservation for accomplishing the overall mission of WRPIS. Positive (symbiotic) associations between WRPIS germplasm (chiefly grasses) and various endophytes, some of which have proven commercially valuable, are also researched and documented (Dugan et al., 2002, Tadych et al., 2007).

The advent of high throughput DNA sequencing technology has generated tremendous amount of DNA sequence information. These include the draft whole genome sequences of two model plant species (Arabidopsis and Brachypodium) and several important crop plants (rice, soybean, sorghum, poplar) and massive numbers of ESTs (expressed sequence tags) of many plant species. This important resource is potentially useful for gene discovery in germplasm since genomics analyses with bioinformatics tools revealed substantial conservation across distantly related species at the DNA sequence level. Technologies are available to assess DNA sequence variation in large numbers of plant accessions to identify different forms (alleles) of a gene. These methods are increasingly robust and speedy, generating data from tiny amounts of plant tissue at any developmental stage unaffected by environmental conditions. DNA-based markers are becoming a key part of plant germplasm management. However, as pointed out by Bretting and Widrlechner (1995), molecular markers should be considered a complement to, not a replacement of, managerial experience with plant germplasm. WRPIS has built a substantial capacity of assessing genetic diversity of germplasm collections with a large number of different molecular techniques that are presently available such as Random Amplified Polymorphic DNA or RAPD (Williams et al., 1990), Amplified Fragment Length Polymorphism or AFLP (Vos et al., 1995), Simple Sequence Repeat or SSR (Diwen et al., 1997) and Target Region Amplification Polymorphism or TRAP (Hu and Vick, 2003). Several projects have been completed in analyzing genetic diversity and relationships among the accessions maintained at WRPIS (Johnson et al., 2002, 2007), in mapping quantitative trait loci (Loridon et al., 2005), in fingerprint the core subsets of pea (Coyne et al., 2005) and in developing markers tightly linked to important economic trait such as disease resistance (Okubara et al., 2002, Coyne et al., 2008). This information will be very useful to geneticists and breeders for genetic improvement of crops.

Morphological characterization of accessions and subsequent documentation also substantially increases the value of germplasm collections. Since the information will be entered into the GRIN database that can be accessed by scientists worldwide, it is necessary to develop and utilize adequate plant character descriptors that reflect national and international standards.

Global agriculture will need to be more productive to meet the nutrition requirements of a growing world population. State-of-the-art technologies that define plant biotechnology will help meet this need. But factors such as climate change, urbanization and environmental degradation are making both genetic resources and arable land increasingly limited. The availability of conserved plant germplasm for research and breeding will be the foundation for sustaining agricultural production into the future. Therefore, the WRPIS with its 80,000 conserved accessions of major American and global crops will assume an even greater role in sustaining 21st century agriculture in the Western Region and other regions of the world.

Related, Current and Previous Work

Ten scientists (four research scientists, one supporting scientist and five curators) are working diligently and collaboratively towards accomplishing the mission of the WRPIS at Pullman, WA. The WRPIS germplasm collection is divided into crop groups and assigned to individual curators. The research scientists conduct mission-related research in agronomy, plant pathology, entomology and molecular genetics to help the curators effectively and efficiently manage the assigned germplasm resources. Curators and the research scientists collaborate on research to solve practical and imminent problems to improve plant germplasm maintenance and management. Since the National Temperate Forage Legume Genetic Resources Unit (NTFLGRU) stationed at Prosser, WA is part of the WRPIS management unit, collaborative efforts are routine between the two sites. WRPIS has frequent interactions with the crop specific NPGS repositories in the Western Region (Aberdeen, Corvallis, Davis, Hilo, Palmer and Riverside, none of which is recipient of W6 funds) through the W6 annual meetings.

Several reports have adequately documented the need for a plant germplasm acquisition and preservation system for the United States (National Plant Genetic Resources Board, 1984, Council for Agricultural Science and Technology, 1985, Janick, 1989, National Research Council, 1991, Qualset and Shands, 2005). These papers describe the components of the U.S. system, including the WRPIS. The only duplication of collections is for security back-up at the National Center for Genetic Resource Preservation (NRGRP), Ft. Collins, Colorado. The NRGRP stores most the NPGS accessions at either -18 C° or under liquid nitrogen (LN, vapor phase). At the WRPIS, we store the original seed samples at -18 C° and the distribution samples, or the active collection, at 4 C° and 28% relative humidity.

Activities WRPIS collections benefit U. S. agriculture by addressing issues of genetic erosion and genetic vulnerability in assigned crops. Accessions in each germplasm collection are documented in GRIN with variable amount of associated data, and the germplasm is freely available for scientific research and evaluation worldwide. There is a wide and diverse customer base for WRPIS germplasm . The user community primarily includes universities and colleges, major commercial seed companies (i.e. Monsanto, Pioneer Hybrid, Syngenta Seeds, Novartis, etc.) plant and animal scientists at international research centers (i.e. CIAT, ICRISAT, ICARDA, AVRDC, IPGRI, etc.), other national programs in many countries, production specialists, farmers, and occasionally home gardeners. The most direct customers are public and private crop breeders that require new genetic traits to improve or develop improved crop cultivars, and university researchers conducting basic research.

There is a wide range of research activity utilizing NPGS plant germplasm within the Western Region. For basic genomics studies, utilization of false brome grass, Brachypodium distachyon, a temperate wild grass species assigned to the WRPIS Agronomy Curator, has increased dramatically. Before 2001, little was known about this grass except variation in chromosome numbers within the genus (Robertson, 1981). Because of its small genome size, rapid growth, self-fertilization, and simple growth requirements, this grass was proposed as a new model for grass functional genomics (Draper et al., 2001). The small genome of 365 mbp makes gene discovery from Brachypodium relatively easier than from grain crops like wheat or barley. Since Brachypodium evolutionarily diverged just prior to the radiation of cereal crops, forage and turf grasses, genes isolated will accelerate fundamental research in food and biofuel crops (Garvin et al., 2008). In the last seven years WRPIS has provided over 1000 seed packets to 95 interested parties in 14 countries working to develop genetic and genomic research resources. An International Brachypodium Initiative (IBI) was formed in 2005 to foster communications among researchers. This collective effort has established Brachypodium as a full-fledged model plant system with a whole range of available genomic resources including i) an efficient transformation system for transgenic study (Vogel and Hill, 2008); ii) an approximate 4X genome coverage draft nuclear genome sequence completed by the U.S. Department of Energys Joint Genome Institute (DOE-JGI) located in Walnut Creek, CA; iii) over 200.000 expressed sequence tags or ESTs (Vogel et al., 2006) and iv) several large insert DNA bacterial artificial chromosome (BAC) libraries ((Foote et al., 2004, Hasterok et al., 2006, Huo et al., 2006). The draft whole genome sequenced line, Bd-21, is derived from PI 254867 from WRPIS. This same line was used for EST and chloroplast genome sequencing (Bortiri et al., 2008).

As an example of germplasm utilization in breeding, the UC Davis lettuce research program developed and released four advanced breeding lines that provide superior resistance to downy mildew in a Salinas horticultural type. This was done by backcrossing wild resistant accessions from the WRPIS collection (PI 491226, PI 491206, PI 491108 and PI 491208) with cv. Salinas as the public domain recurrent parent. The resulting lines expressed resistance to all Californian isolates. These sources of downy mildew resistance have not been used as parents for existing cultivars grown in California and therefore provide new downy mildew resistant genes. In related work, Hayes and McCreight (2004) identified two leaf lettuce accessions, PI 289042C (cv. Ausztraliai Sarga) and PI 289035C (cv. MayKing) showing resistance to lettuce big vein, a serious virus disease in both California and Arizona. Dr. Phil Bruckner, Dept. of Plant Sciences and Plant Pathology, MSU-Bozeman, received six Triticum aestivum accessions. PI377884, Gatcher, was acquired because it is reported to be susceptible to root lesion nematodes. It was used in basic research on this pest in Montana. Three accessions of Mara (PIs 244854, 259890, and 292756) were acquired as a potential source of reduced height gene Rht8. The gene was incorporated into Montana germplasm from an alternative source. Two accessions of Vernopolis (PIs 214401, 297008) were acquired as potential sources of resistance to wheat leaf blotch, a foliar disease caused by Septoria tritici. Several other germplasm sources obtained from NPGS are being used as sources for resistance to Russian wheat Aphid in current breeding efforts. Heather Cook of Butte, MT evaluated numerous vegetable germplasm of tomato, pepper, bean, garlic and cucumber accessions provided by NPGS for local adaptation and is hoping to breed locally adapted varieties to encourage local production of vegetables for local consumption and sale. Dr. Martin at Montana State University has evaluated the agronomic performance of Camelina germplasm obtained from the National Plant Germplasm collection (NC7) for their potential as a new oil seed crop in Montana and other states in the Northern Great Plains. Dr. Ray at New Mexico State University and Dr. Groose at Wyoming State University used many alfalfa germplasm accessions from WRPIS to develop better forage crops with high water efficiency and improved profitability for farmers growing alfalfa forage. Scientists at Utah State University utilized plant materials from WRPIS in the development of released cultivars of meadow brome, Altai wildrye and Russian wildyre.

WRPIS germplasm is exceptionally useful for specialty crops. For example, the plant breeding programs at OSU are mostly centered on crops like hazelnuts, hops, and meadowfoam, that have little private sector breeding activity. Dr. Shawn Mehlenbacher used the Corylus germplasm extensively for assessing genetic variability, developing DNA markers associated with desirable horticultural traits and breeding new cultivars for Oregon's hazelnut industry by using DNA marker-assisted selection for disease resistance. Other users of NPGS in Oregon include federal researchers as well as private seed company and individual breeders.

University of Arizona used germplasm from the NPGS in numerous research projects that included the genetics of crop plants and the domestication of new crops, especially in research involving the evaluation of the genetic diversity found within the germplasm of new and/or under-utilized crops suitable for cultivation in arid and semiarid environments. For example, Dr. Dennis Ray conducted research to elucidate the basic biology guayule, a potential domestic source of natural rubber, for not only increasing rubber yields, but also studying its rather complicated reproductive biology. In Montana, Dr. Martin and colleagues used the barley core collection from National Small Grains Collection in his research to further the understanding of the hardness locus in barley and its impact on grain quality traits. Several research programs associated with the University of Idaho used numerous NPGS germplasm. Accessions of cereals, beans, rapeseed, mustard and potato have been used in breeding and to develop new cultivars with improved biotic stress resistance, abiotic stress tolerance/resistance and improved or modified end-use quality. Included in this research are Dr. Zemetras efforts to map QTL and develop molecular markers to assist in the transfer of any desired traits identified into adapted genotypes.

Germplasm has been used in programs associated with research in entomology and plant pathology to study the mechanisms of resistance/tolerance to various insects and diseases. The ARS-led National Sclerotinia Initiative aims at neutralizing white mold's economic threat to seven different crops: sunflower, soybean, canola, edible dry beans, chickpeas, lentils and dry peas. The WRPIS curators have been providing accessions to the funded projects for evaluating for resistance (Dr. Weidong Chen, ARS-Pullman working on lentil and chick pea, Dr. Phil Miklas, ARS-Prosser, WA working on common bean and pinto bean); for map of QTL for white mold resistance (Dr. Mark Brick, Colorado State University working on dry bean) and for pyramiding and introgressing white mold resistance (Dr. Shree Singh, University of Idaho and Dr. James R. Myers, Oregon State University, both working on pinto and common bean).

Summary of CRIS search with Objective key words:
Three records for "Conserve & Regenerate" and "Priority Plant Germplasm" were found, one of which is an expired CRIS and one the current CRIS for WRPIS. The third project places emphasis on secondary back-up, but only arid-land native species. A search of "Allium sativum" and "long-term storage" produced 2 records; the current CRIS for the WRPIS, and one for the NCGRP. This work is not duplicated in any other project.

No records on pollination of Glycyrrhiza species were found. One 1988 record for insect-Glycyrrhiza interactions, which involved seed predation by a bruchid beetle, was located, but no active records for U.S.-based research on eriophyiid (Aceria spp.) mite infestations and damage on garlic leaves or bulbs. A search of Neotyphodium grass endophyte records produced a total of 49 records, but no active U.S.-based research on incidence of viable endophytes in grass seed in germplasm repositories and influence of diverse grass  Neotyphodium associations on aphid performance. None of the Neotyphodium records identified active research on the response of Rhopalosiphum padi to different wild grassendophyte associations in the WRPIS seedbank expressing different alkaloid profiles. A search identified 17 active projects on cereal leaf beetle, Oulema melanopus, but no active projects duplicate ongoing research proposed in this project.

Alternaria - There are CRIS projects pertinent to different plant species but none to Alternaria from grass family hosts. There are also CRIS projects 0192518 (Pryor, primarily tree nuts, but he has some WRPIS isolates too) and 0180234 (Peever, predominantly citrus isolates, but he has some of our grass isolates, too). WRPIS is collaborating with Pryor (on A. infectoria) and with Peever (on A. alternata and A. tenuissima). There are no CRIS projects duplicating WRPIS efforts.

Cladosporium - There are CRIS projects pertinent to food-borne fungi (0207658 E.E. Rico), air-borne allergens and indoor air (0201979 D.W. Li) and Pacific Northwest Fungi (0194100 Glawe et al.). WRPIS assists the latter project by identification of Cladosporium isolates and coauthoring of manuscripts, but the Glawe et al. project does not duplicate the WRPIS project, which concentrates on seed-borne Cladosporium germaine to NPGS germplasm. The WRPIS project is the only one working on Selenophoma (Pseudoseptoria) and the only project involving Aureobasidium and Cicer is at WRPIS.

Records showed three safflower research projects, but only Acc. No. 0174701 "Breeding and improvement of oilseed crops for eastern Montana" is related, and the leader of this project is a WRPIS cooperator, sharing information and germplasm, so there is no duplication. No projects other than the WRPIS are working on genetic enhancement of winter safflower.
As indicated in the search above, none of the grass genera listed are being collected and characterized as we propose. The project closest is Acc. No. 0404857 at Logan UT, which has been a long-term cooperator with the WRPIS in both collection and evaluation research. Nether duplicate collections or research will be pursued, only collaboration. The work proposed should strengthen the opportunities for breeding better adapted native plants by the Logan ARS Unit and others involved with developing native germplasm.
Each genus listed in Objective 4 was searched for pertinent molecular and phenotypic characterization projects. The number of research projects ranged from 26 for Beta to zero for Carthamus, Vicia, and Lomatium (except for the current WRPIS project). Projects identified with potential overlap were those which already cooperate extensively with WRPIS. These include projects on food legumes (Acc. No. 0405211 Enhanced collaboration on genetic resources and the use of wild relatives in Chickpea; Acc. No. 0405363 Control of bean and pea diseases through germplasm enhancement and improved cultural practices; and Acc. No. 0406527 Improved plant genetic resources for pastures and rangelands in the temperate semiarid regions of the Western US). Completion of the proposed research will strengthen rather than duplicate other research projects involved in germplasm enhancement and breeding of these species. A search on plant association mapping studies resulted in 242 projects, none on association mapping populations in pea, lentil or chickpea.

A search of "Plant Germplasm Acquisition" revealed 16 projects. Most of these were complementary plant germplasm programs within or supporting overall NPGS programs. There are no comparable projects in the U.S. For genetic stocks of pea and bean a search indicated no other acquisition project for pea and bean in the USA.

Objectives

1. Conserve and regenerate priority cool season food and forage legume, beans, turf and forage grass, native rangeland, oilseed, vegetable, ornamental, medicinal, and other specialty and industrial crop genetic resources, and distribute samples and associated information worldwide.
   
2. Refine and establish regeneration protocols for efficiently and effectively regenerating insect-pollinating germplasm accessions of various crop groups; monitor potentially pathogenic microorgamisms such as fungi and viruses, and ensure the health of the germplasm collection.
   
3. Evaluate priority crop core subsets and other selected germplasm with morphological descriptors, and key agronomic or horticultural traits, such as general adaptation, phenology, and growth potential. Identify accessions with desirable economical traits for multiple location tests and potential release to broaden the genetic base of breeding gene pools.
   
4. Apply molecular marker techniques to assess diversity, detect duplicated accessions, identify taxa that were difficult to classify with morphological characteristics and associate DNA polymorphism with variations of important economical traits in selected crops.
   
5. Expand strategically the genetic diversity in the WRPIS collection and improve associated information for priority cool season food and forage legumes, beans, turf and forage grasses, native rangeland plants, oilseed, vegetable, ornamental, medicinal, and other specialty and industrial crop genetic resources.
   
6. Promote the use of the diverse germplasm collections in the NPGS to reduce crop genetic vulnerability and sustain crop production within the Western Region, within the United States and throughout the world, employ different avenues of technology transfer in the form of the Internet-accessible database, research publications and professional conferences to encourage information sharing among scientists and coordinate plant germplasm distribution to researchers worldwide for future crop development.
   

Methods

1. During regeneration, the biggest challenge is to maintain the genetic integrity and diversity of the original collection. We have been developing practical regeneration protocols that are tailored to the specific needs of a given species as well as our available resources. For insect-pollinated taxa, various approaches have been integrated into the regeneration protocols with a careful consideration of pollination biology (Clark et al., 1997; Johnson et al., 2004; Clement et al., 2007a, 2007d). For highly self-pollinated species, such as most of WRPIS food legumes, the main considerations relate to plant health. Control measures are employed to ensure pest and disease free seed production using chemical or cultural control with the contribution of our entomologist and plant pathologist. For example, fine screening is placed over ventilation windows in greenhouses in which bean accessions (Phaseolus vulgaris) are regenerated to prevent entry of the aphid vector for seed borne Bean Common Mosaic Virus.

We will continue to use screen caging for regenerating insect pollinated species, such as Allium and most forage legumes and optimal insect pollinators (Clement et al., 2007a, 2007d). Caging is also effective in preventing intercrossing among accessions when grown in the same field for some species such as safflower (Carthamus tinctorius), that is generally self-pollinating (Johnson and Li, 2008). Most WRPIS grasses are highly heterogenic and wind pollinated, so distance isolation of at least 50 m and plant populations of over 100 plants per accession is desirable to limit unintended outcrossing and to slow genetic drift (Johnson, 1998; Johnson et al., 2004). Research using molecular markers is ongoing to determine how various types of seed samples affect allele frequency and diversity in accessions during regeneration (Johnson et al., 1996; Johnson et al., 2006). Ongoing collaborative work at the WRPIS between the entomologist, agronomist, and curators is aimed at developing and refining effective insect pollination systems using screen cages. Protocols using geographic isolation, pollen proof cages, insect containment cages and insect exclusion cages are utilized for out-crossing species

We will continue the greenhouse increases of Phaseolus using pot culture, which allows continuous cropping in each row and also facilitates virus testing on an individual plant basis and hand pollinating wild Phaseolus species to increase seed set. Pest control will continue to be based on parasites and predators with chemical treatments confined to spot applications of low mammalian-toxicity materials.

We will continue our emphasis on germination tests which guide regeneration needs and ensure high quality seed distribution. We will determine the viability of 11,000 accessions over the next 5-year period by performing replicated viability tests using guidelines in the Association of Official Seed Analysts "Rules for Testing Seed", the International Board for Plant Genetic Research publication "Handbook of Seed Technology", and other sources as needed. Since resources will not allow completion of viability testing on all seed samples, priorities will vary depending on the species and the situation. When appropriate and available, NCGRP viability testing data will be used.

A high priority will be given for regeneration to the 12,025 accessions of original seed packages that have never been increased. These include Allium, various special purpose legumes, Vicia, and certain miscellaneous and native species and represent approximately 15% of total WRPIS holdings. Our goal is to make most of these accessions available for distribution and back-up during the next funding period.

The WRPIS active seed collection is stored at an established optimum of 4°C with 30% relative humidity. We receive seed orders almost every day from the germplasm user community worldwide through e-mail, fax, letter, or telephone. Seed orders are filled in a timely manner, usually within a week, and distributions are documented in the GRIN database.

Currently, 74% of available WRPIS accessions are securely backed up at NCGRP or another NPGS site. But there is a large variation among collections. For example 95% of the Lens collection is backed-up, and 93% of the Carthamus and Medicago, but the percentages for Allium, Vicia, and special purpose legumes are 23, 42, and 44 %, respectively. Special effort will be concentrated on those collections below the 70% back-up benchmark to ensure that those accessions at highest risk of loss are incorporated into regeneration cycles to produce seeds for back-up and distribution. For garlic, an additional 100 accessions will be cryopreserved at the NCGRP using the methods of Volk et al. (2004).

2. We will conduct pollination research on licorice, Glycyrrhiza glabra and G. lepidota, two key medicinal species maintained at the WRPIS. First we will determine the degree of selfing or any pollinator requirements. Plots will be established at Pullman and Central Ferry, WA sites, where the presence of diverse insect pollinator communities has been established (Clement et al., 2006; Clement, unpublished). Two single-row plots will be established in spring 2009 at each site, with 20 plants of each species per plot (two species and two accessions per site). Plots at each site will be separated by at least 150 m. The breeding system of each species and accession will be studied in the field, comparing pod and seed set in a series of bagged flower clusters (where only self-pollination is possible), and non-bagged control flower clusters (open to natural pollination). Fine nylon mesh material will be used first, but pilot studies with different fabrics may be required to identify the best bagging material (Neal and Anderson, 2004; Degrandi-Hoffman and Chambers, 2006; Wheelwright et al., 2006). Second, we will identify potential pollinators in 2009 and 2010 with the established methods (Clement et al., 2006). The bee diversity information will identify potential pollinators for cage pollination trials on Glycyrrhiza in 2011 and 2012 to verify that insect pollinators are required for good pod and seed set. Similar approaches will be extended to other germplasm species for refining and establishing regeneration protocols.

We will examine new germplasm entering collections to ensure freedom from pests. Seeds frequently constitute an important means of movement for pests and pathogens McGee, 1997). We will monitor regeneration plots for pests and pathogens. To reduce the spread of viruses, seed borne in several crops, we will continue our virus testing and eliminating effort. Depending upon the crop, and since there is rarely 100% seed-to-seed transmission of seed borne viruses, protocols are used to both produce virus free distribution samples and maintain the genetic diversity within an accession (Klein et al., 1986 and 1987).

We periodically survey garlic germplasm for fungi and viruses using standard protocols for isolation and/or identification (Mueller et al., 2004; Dugan, 2006; Pappu et al., 2005, 2006). With regard to fungi, these protocols can be summarized as comprising i) obtaining germplasm, ii) incubating cloves at conditions permissive for fungal growth (elevated humidity at ca. 25ºC) over mineral oil (oil prevents transfer of mites between test lots), iii) transfer of fruiting bodies and/or symptomatic tissues to agar media appropriate for the target fungal taxa, iv) incubation under periodic florescent + near ultraviolet light for promotion of sporulation, v) identification to genus and species by literature specified in the above publications, and vi) preservation of representative fungal isolates in glycerol in liquid nitrogen, in a low temperature freezer, and/or other methods in accordance with our standard operating procedures. Pathogenicity of taxa not previously well documented as pathogenic to garlic is tested by standard fulfillment of Koch's postulates using appropriate controls, and in the case of Fusarium oxysporum f. sp. cepae (morphologically indistinguishable from saprophytic F. oxysporum) including a positive control obtained and stored under USDA-APHIS PPQ permit.

With regard to garlic viruses, protocols can be summarized as i) survey of WRPIS-NPGS field plots for symptomatic plants, ii) using commercial anti-sera (Agdia, Inc., Elkhart IN) for detection of Garlic common latent virus, Iris yellow spot virus, Leek yellow stripe virus, Onion yellow dwarf virus, Tobacco rattle virus, and Tomato spotted wilt virus, with ELISA results to be verified by PCR using virus-specific primers targeting the ITS (internal transcribed spacers) following the published protocol (Weir et al., 1990). Results to date have been published (Pappu et al., 2008).

Powdery mildews inhibit regeneration of cool season legumes, and there is evidence that the accepted taxonomy and host-fungus data for powdery mildews on cool season legumes are obsolete. Our collaborator, Dr. W. Chen, a USDA-ARS plant pathologist will conduct the DNA laboratory work of amplifying ITS. The Bio101 FastDNA kit will be used for DNA extraction and the sequencing of the amplicons will be performed with a PE Biostems Model 377 DNA Sequencer. Finally, a BLAST search will be conducted according to (Altschul et al., 1990). For expanded ITS, sequences will be cloned into a PCR 4-TOPO (Invitrogen) vector. Drs. Dugan and Glawe will assist with correlation of DNA sequence data with morphological data described by Braun (1987) and Glawe (2006). Preliminary results have been published (Attanayake et al., 2008).

We will assay selected stored seed accessions for viable microbial associates. Methods for recovery and identification of fungal taxa will be largely from Mueller et al. (2004) and Dugan (2006) as summarized under sub-objective 2C, but with the accessions surveyed to be established in collaboration with the curator of that germplasm and with reference to specific problems encountered by the curator (e.g., pre- and post-emergence damping off in lupine seed will necessitate media selective for Pythium as well as media for anamorphic ascomycetes). Taxon-appropriate media are documented in Mueller et al. (2004), Stevens (1981), and multiple other sources. Koch's postulates will be conducted if taxa appear pathogenic but not previously documented as such.

Surveys of WRPIS crops and alternative weedy hosts will be monitored for diseases in the field, and pertinent taxa identified by standard methods (Mueller et al., 2004; Dugan, 2006; Pappu et al., 2005, 2006) and, when necessary, comparisons of ITS and/or other sequences with data in GenBank. Special emphasis will be placed on under-reported taxa such as powdery mildews, rusts or downy mildews on alternative weedy hosts.

3. Approximately 3,700 accessions will be evaluated for basic descriptors in the field, screen house, greenhouse and laboratory during the next five years. This will result in approximately 57,250 data points of phenotypic evaluations and 1,550 images to be uploaded to the GRIN database. Established descriptors are given in GRIN with detailed explanations. Briefly, all Phaseolus accessions grown in the greenhouse will be evaluated for photo-response, emergence type, hypocotyl color, leaf shape, days to flowering, flower concentration, wing and vein characteristics, pollination requirements, growth habit, pod characteristics, root type, and seed characteristics. On 30 Allium plants per regeneration plot, anther color, day of first flower, flower color, flowers/umbel, umbel diameter, days to seed maturity, and several leaf and scape characteristics will be recorded. On a target population of 100 Beta plants per accession, field grown plants will be evaluated for accession uniformity, plant habit, bolting tendency; cluster fasciation, life form, flower pattern between plants, flower pattern within plant, hypocotyl color, and several leaf characteristics. On a target population of 15 Lactuca plants per field grown accession, plant height, flower color, flower diameter, seed color, days to bolting, days to flowering, days to seed maturity, and leaf spines will be taken. Pisum descriptors will include bloom day, days to flower, flower and pod characteristics, plant height, leaflet morphology, and stipules. Similar descriptors will be taken for Cicer and Lens. Carthamus regeneration plots will be evaluated for fresh and dry flower color, spininess, plant height, head size and shape, bloom data, and habit.

Digital images will be taken to document accessions in cool season grasses, Allium, and Carthamus field-nurseries. For Lens and Cicer core collections, digital images will be generated using a flatbed scanner and software designed to automatically measure seed characteristics. The Carthamus descriptors will be updated for seed oil fatty acids and new descriptors for vegetative structure added to the cool-season grass descriptor list.

We will select individuals from accessions with desirable traits such as disease resistance, insect tolerance and abiotic stress tolerances during evaluation for multi-location tests with our collaborators following the established field experimental procedures. After confirming the expression of the potential useful traits, the selected accessions will be released to public to promote germplasm ultilization.

4. We will pursue molecular characterization of collections, core collections, or other subsets of Achnatherum, Allium, Pisum, Medicago, Bromus, Poa, Vicia, Lomatium, Lens, and Phaseolus. Numerous molecular marker systems are available to identify phylogenetic associations among accessions (Williams et al, 1990; Vos et al., 1995; Balloux and Lugon-Moulin, 2002; Vieux at al., 2002; Hu and Vick, 2003). The objective of a given analysis will guide the use of the available marker systems. For example, population screening for candidate genes might best be served using SSR derived from annotated expressed sequence tags (ESTs). For assessing genetic diversity and identifying population structure, AFLP and TRAP are more cost-effective.

We have established laboratory procedures for various types of molecular markers for the proposed research projects. The updated protocol (Johnson et al., 2007) will be followed for DNA extraction. For AFLP marker assays, the basic procedure (Vos et al., 1995) will used. The selective amplification is modified as a 10¼l reaction, including 0.25 Units of Taq polymerase and the accompanying buffer, 2 ¼l of MseI primer from Life technologies kits, 0.5 ÁMoles of fluorescent-labeled EcoRI primer and 2 ¼l of preamplified DNA diluted 10:1 with 0.1X Tris buffer. For TRAP marker amplification, the published protocol (Hu and Vick, 2003) will be used. The amplification is in a 15¼l total volume, including 0.15 Units of Taq polymerase and the 1X of accompanying buffer, 1 pMol of fixed primer, 0.3 pMoles of fluorescent-labeled arbitrary primers and 2 ¼l of DNA diluted to 10 ng ul-1. For SSRs, methods of Balloux and Lugon-Moulin (2002) are followed. A reaction volume of 10¼l is used with 0.1 Unit of Taq polymerase; 25·g of template DNA, and reagent concentrations of 150¼M for each dNTP, 1.5mM MgCl2, 0.2¼M each of forward and reverse primers. The amplification method starts with an initial denaturing step at 94°C for 30s, followed by 15 cycles beginning at 94°C /10s, 65°C /30s, 72°C/30s and stepping down the annealing temperature one degree each cycle to 50°C , and ending with 10 cycles of 94°C /10s, 50°C /30s, 72°C /30s. Separation and visualization of AFLP, TRAP and SSR markers are done on 6.5% polyacrylamide using a Li-Cor DNA Analyzer. Gel images are printed and scored manually.

Numerous software applications for phylogenetic analysis of germplasm collections are available. Measurements of within to among population genetic variation are also available using various molecular markers (Nei, 1973; Culley et al., 2002). Software packages such as NTSys-pc (Rohlf, 1992) and Clustan® (Clustan Ltd, Edinburgh, Scotland) provide phenographic associations of population relationships as well as Principal Component or Principal Coordinate Analyses. Estimates of the number of discrete or admixed populations represented in the analysis will be estimated using allele correlations by the software STRUCTURE (Pritchard et al., 2000), with the most probable k-clusters determined as given by Evannos et al. (2005).

We will perform association mapping in three crops on a selected sample of 384 accessions of pea, lentil and chickpea. SSR genotyping data, disease reaction and yield component data from field studies replicated over environments will then be used to uncover markers associated with functional candidate genes. Population structure will be estimated on the basis of unlinked SSR markers. The extent of linkage disequilibrium (LD) will be estimated relative to the LD observed among unlinked markers. Association of SSR loci with yield component and disease resistance will be analyzed through a mixed-effects model, where subpopulation was considered as a random factor and the marker tested was considered as a fixed factor. Permutations will be used to adjust the threshold of significance for multiple testing within linkage groups. Previous QTL analysis, significant markers for disease and/or yield components will be tested and alleles potentially useful for selection may be identified.

5. We will continue to acquire new germplasm to fill the existing gaps in our current collections of Beta, Cicer, Lens, Pisum, Vicia, Lathyrus, Salvia, native grasses and forbs via plant exploration/collecting and germplasm exchange. Exploration proposals will be submitted to the USDA-ARS NPGS Plant Exchange Office for collection of both foreign and domestic species (Allium, Beta, Cicer, Lens, Pisum, Vicia, Lathyrus, and Poa). Exchange and cooperative collection of native species are being pursued within ARS and cooperatively with other Agencies such as the Natural Resources Conservation Service, the Forest Service, and the Bureau of Land Management (Achnatherum, Poa, Camassia, Balsamorhiza, and Lomatium). Essential in acquisition is passport data including GIS coordinates, sound taxonomy, and a physical and botanical description of collection locations including digital photographs. Additionally, exchanges will be initiated with foreign plant germplasm repositories (Cicer, Lens, Pisum, Vicia, and Lathyrus) especially through the CGIAR centers.

Priorities for the acquisition of genetic stocks will be determined by the curators working with the CGCs. For each crop and corresponding model plant system, the needs of the research community will be assessed and current genetic stocks in the NPGS and other gene banks inventoried. The inventories will determine gaps in the NPGS genetic stocks collections. The availability of the genetic stocks to researchers and the role of the WRPIS will be determined. WRPIS curators will acquire and characterize new genetic stocks through exchange or donations, depending on specific crop needs. The work over the next five years will focus on four crops: bean, pea, chickpea, and lentil. The primary source of new genetic stocks will be from the published literature. For bean, the main source of published genetic stocks is the Bean Improvement Cooperative Newsletter and the Journal of Plant Registrations. For pea, chickpea, and lentil, the main sources of published genetic stocks are Pisum Genetics Newsletter, Journal of Plant Registrations, and the International Chickpea and Pigeonpea Newsletter.

We strongly believe that a cooperative approach is best in solving problems in agriculture of the Western region, which comprises diverse environments and crop species. The proposed project was developed and will be conducted under the guidance of our Technical Advisory Committee with members in multiple disciplines and from each of 13 Western States. Members of this Committee are experts in areas such as statistics, crop genetics and breeding, agronomy and plant pathology. Most participants from the current project (2003-2009) will continue to play important roles in carrying out the project. Strong collaboration has been developed among the participants through the past years. The projected body of work for the next 5 years will ensure that valuable plant germplasm resources (and associated data) are available, preserved in a superior condition, and can be accessed expeditiously by our customers into the future.

6. We will continue to promote the use of the diverse germplasm collections in the NPGS by employing all available communicating means Internet-accessible database, research publications and professional conferences to encourage information sharing among scientists. It has been known that the genetic vulnerability of modern high-yielding varieties of major crop species in stemmed from genetic uniformity. Plant breeders in the Western Region, in the United States and throughout the world, have been using the available plant germplasm to reduce crop the genetic vulnerability. This project will encourage and monitor the germplasm utilization in the Western states. Germplasm related data on Plant Introduction accessions in the NPGS reside in the Germplasm Resource Information Network (GRIN) data base. All data designated as available to the public is accessible via the Internet. GRIN is the primary repository for passport, evaluation and characterization data related to WRPIS germplasm. Germplasm is received by the WRPIS as material transferred from other NPGS sites, from NCGRP, through exchanges with other national genetic resource conservation programs around the world, from public and private sector breeding programs, from donations by U. S. citizens, and from plant exploration expeditions. Passport data is assembled at Pullman and loaded into GRIN. Research and evaluation data not yet in GRIN is maintained and distributed by the respective scientists in charge of the projects. Information and results from research projects is transferred to the user community via peer-reviewed publications in scientific journals, posters, abstracts and presentations at the international, national and regional scientific and commodity meetings. These publications and presentations lead to personal interaction with scientists from around the world and potential germplasm exchange.

Measurement of Progress and Results

Outputs

• Output 1: This project will continue to provide quality germplasm of the species maintained at this site to researchers worldwide. The utilization of this germplasm in basic research will result in strengthening of plant sciences and genomics by documenting genetic variation, plant-environment interaction, linkage maps of useful genes and DNA sequence information. Breeders in applied research will incorporate novel genes into locally-adapted cultivars with enhanced pest resistance, improved end-user quality and increased productivity. Planned experiments will lead to development of new crops for industrial, ornamental and medicinal purposes.
• Output 2: Efficient regeneration protocols will be established and refined based on the research information on pollination biology for effective regeneration of the respective species and accessions in the WRPIS collection. Potentially beneficial fungi (endophytes) will be maintained and harmful fungi, viruses, and other pathogenic microorganisms as well as pests will be managed during regeneration to produce healthy seeds for storage and distribution.
• Output 3: More phenotypic data associated with priority accessions will be available to the US and worldwide breeders and researchers who use our germplasm. The data include digital images, morphological descriptors, and important agronomical or horticultural traits such as disease and insect resistance, general adaptation and growth habit. All the data collected will be uploaded to the GRIN database that can be accessed by the public through the Internet. Selected germplasm will be released for use in breeding programs.
• Output 4: Our molecular characterization program will generate information on molecular diversity and population structure of selected crop species and wild relatives. Application of this information to germplasm management will increase our overall efficiency and effectiveness by eliminating duplicated (redundant) accessions, identifying genetic gaps in the collection and monitoring allele frequencies to maintain genetic integrity during regeneration. Information on DNA markers associated with economic traits will be published in peer-reviewed journals, enabling breeders to incorporate novel traits into elite lines via marker-assisted selection in their cultivar development efforts.
• Output 5: One thousand more accessions of priority crop species will be added to the WRPIS collection for distribution. These new accessions will fill the existing gaps in our collection as revealed with DNA-based markers, morphological variation or geographical origin. The acquisition will be accomplished by collection trips and germplasm exchanging program through WRPIS personnel and their international collaborators.
• Output 6: The utilization of the NPGS germplasm accessions in the crop genetic improvement research will result in numerous scientific publications. It will lead to the release of breeding lines or crop varieties with better resistance to plant pests and pathogens, more tolerance to abiotic stresses and improved ender-use qualities. It will also make possible the development of new crop for specific market.

Outcomes or Projected Impacts

• US breeders and researchers will have additional accessions of crops, crop varieties, and native plant genetic resources for host-plant resistance and value-added nutritional traits.
• Our collections will promote continued genetic improvement of important agricultural crops and restoration of public and private lands.
• Accessions of priority plant genetic resources will be secured and the genetic gaps in the collections will be filled through acquisition.
• Germplasm will be more efficiently and effectively conserved, monitored for seed quality and health, and distributed on request worldwide.
• Methods will be developed and refined for regenerating germplasm collections.
• Accessions of priority genetic resources will be evaluated ("phenotyped") for key traits related to adaptation, yield components, and host-plant resistance to diseases and insects.
• Diseases and their etiological agents will be identified and characterized from selected crops and native plants.
• Insect pests will be identified and characterized from selected crops and native plants from the western region. Genotypic and phenotypic (evaluation) datasets for key genetic, agronomic, and/or horticultural traits will be incorporated into GRIN and/or other databases, thereby expanding worldwide access to critical data.

Milestones

(0):m 2010 to 2014, and each and every year, we will regenerate 1,500-2,000 priority germplasm accessions including 500 cool season food legume, 400 beans, 300 grasses, 250 Alliums, 40 ornamentals, 30 safflowers and 20 Beta for Objective 1; and collect descriptor data and images on 500 regenerated accessions and upload the data into GRIN for Objective 3; and use 5,000 germplasm packets from the NPGS for crop improvement in the Western States for Objective 6.

(2010): For Objective 2, we will report viral and fungal pathogens of garlic; differentiate Erysiphe species occurring on cool season legumes in the Pacific Northwest and test effectiveness of application of suspensions of Aureobasidium pullulans to post-harvest chickpea debris as a biological control for Ascochyta blight of chickpea. For Objective 3, we will complete an initial field screening for winter hardiness of 100 faba bean accessions in Pullman and Central Ferry. For Objective 4, we will complete molecular diversity analysis for the pea core with15 SSRs, the chickpea core with 10 SSRs and publish the data; compare TRAP and Barcode sequence evaluation for Allium species determination; analyze diversity and redundancy in the Allium cepa collection; optimize bulked plant chloroplast marker evaluation of Medicago sativa; and assess diversity within and among populations of Brachypodium distachyon. For Objective 5, we will acquire additional 240 pea genetic stocks from John Innes Centre germplasm collection to increase the genetic diversity of pea genetic stocks.

(2011): For Objective 2, we will complete surveys of grass accessions for viable Neotyphodium endophytes; refine eriophyiid mite sampling protocol for garlic bulbs and commence sampling to quantify mite infestations; report the completed mite and Cereal Leaf Beetle research; assay for resistance in garlic and/or related Allium spp. to Penicillium decay; and report powdery mildew pathogens of cool season legumes in the Pacific Northwest. For Objective 3, we will report the results of winter hardiness of 100 faba bean accession in Pullman and Central Ferry; phenotype RIL and advanced breeding lines for resistances to pea root rots. For Objective 4, we will collect data on chloroplast marker evaluation of Medicago sativa; fingerprint Poa secunda and Vicia arietinum for studying diversity within and among populations. For Objective 5, we will import 70 Chinese landraces from Australia genebank to increase genetic diversity of faba bean collection and omplete phenotype data analysis, publication, and data entry to GRIN for pea core and chickpea core.

(2012): For Objective 2, we will establish field plots for caged pollination of Glycyrrhiza; refine experimental procedures for resistance to Penicillium in Allium, expand trials, incorporate larger numbers of accessions; and report Aureobasidium pullulans as a biological control for Ascochyta blight of chickpea. For Objective 3, we will expand the field screening for winter hardiness of 200 faba bean accessions in Pullman and Central Ferry; and phenotype RIL and advanced breeding lines for resistances to pea root rots (with collaborators). For Objective 4, we will fingerprint Lomatium dissectum, and Cicer arietinum for studying diversity within and among populations and initiate pea RIL QTL mapping. For Objective 5, we will import the international composite collection from ICARDA to increase the genetic diversity of lentil.

(2013): For Objective 2, we will conduct field cage experiment to evaluate pollinators for seed production of Glycyrrhiza accessions; complete sampling of garlic for mites and evaluate results to design control program; report mite and Cereal Leaf Beetle research; conduct large scale, replicated trials with multiple sources of Allium germplasm for resistance to Penicillium decay. For Objective 3, we will initiate developing winter hardiness faba bean lines; and phenotype RIL and advanced breeding lines for resistances to pea root rots. For

Projected Participation

View Appendix E: Participation

Outreach Plan

We will keep updating the Germplasm Resource Information Network (GRIN) database with related data on germplasm accessions maintained in WRPIS. GRIN is the NPGSpublic database that can be accessed from anywhere in the world via the Internet. We utilize GRIN as the primary repository for passport, evaluation and characterization data. The passport data include the taxonomic name and the origin of the germplasm accession (such as being received by the WRPIS as material transferred from other NPGS sites, from NCGRP, through exchanges with other national genetic resource conservation programs around the world, from public (universities primarily) and private sector breeding programs, from donations by U.S. citizens, and from plant exploration expeditions). Passport data is assembled at Pullman and loaded into GRIN. Our scientists/curators will transfer information and results from research projects to the user community by publishing papers in peer-reviewed publications, presenting poster and oral presentations at professional conferences and commodity meetings. These publications and presentations lead to personal interaction with scientists from around the world, and subsequent additional technology transfer occurs.

WRPIS curators serve as committee members or chairs of the respective national Crop Germplasm Committees (CGC) and other academic or social organizations. To mention a few, the WRPIS Agronomist is the Chair of the International Safflower Germplasm Committee, Member of the Technical Advisory Committee for the Special Grant, Grass Seed Cropping Systems for Sustainable Agriculture, active ex-officio member of the Forage and Turf grass CGC; the WRPIS Horticulture Curator is the Ex-officio member of six CGCs (Root and Bulb, Leafy Vegetable, the Herbaceous Ornamental, New Crops, the Clover and Special Purpose Legume and sugar beet) and the member of two PGOC subcommittees (Medicinal Plant and In Situ Conservation); the WRPIS Agronomy Curator is the Ex-officio member of Forage and Turf Grass CGC (Descriptor Subcommittee Secretary) and New Crops CGC; the WRPIS Cool Season Food Legumes Curator is the Ex-officio member of the Food Legume CGC and the member of the Plant Germplasm Operations Committee; and the WRPIS Phaseolus Curator serves as member in for organizations (Phaseolus CGC, Bean Improvement Cooperative Genetics Committee, W1150 Regional Project and Seed Savers Exchange). By participating the regular meeting and other activities of these organizations, we effectively outreach and interact with our stakeholders, customers and general public. We provide updated information and technology documented in our operations manual to requestors from other genetic resource conservation organizations within the US and from other countries and international conservation institutes.

NPGS requires us to distribute available germplasm to requestors worldwide in a timely manner. We routinely fill and ship out the samples within 7-10 days for all regular requests. We also request feedback from the recipients to improve our service to meet the needs of germplasm users in the scientific community.

Organization/Governance

The recommended Standard Governance for multi-state research activities includes the election of a Chair, a Chair-elect, and a Secretary for the Technical Advisory Committee. All officers are to be elected for at least two-year terms to provide continuity. Administrative guidance will be provided by an assigned Administrative Advisor and a CSREES Representative. Over the next five years we will use internal benchmarks and accountability systems to assess progress and then determine future needs. In addition to the input from the W6 Technical Advisory Committee, we will use the ARS, National Program 301 review process, input from the CGCs, and if appropriate, suggestions from and external review.

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Attachments

Land Grant Participating States/Institutions

CA, CO, ID, MT, NC, NM, OR, WA

Non Land Grant Participating States/Institutions

ARS-WA, USDA-ARS/Washington, Utah