W6: Maximizing Conservation and Utilization of Plant Genetic Resources and their Information

(Multistate Research Project)

Status: Active

W6: Maximizing Conservation and Utilization of Plant Genetic Resources and their Information

Duration: 10/01/2021 to 09/30/2026

Administrative Advisor(s):


NIFA Reps:


Non-Technical Summary

Statement of Issues and Justification

The need, as indicated by stakeholders:



Humanity cannot survive without agriculture. Improving agricultural efficiency and sustainability will play a vital role towards improving global food security considering global population growth and projected negative impacts affiliated with climate change. To meet this challenge, plant breeding and other plant science research fields rely on the diversity of discoverable traits and genes in agriculturally important plant genetic resource (PGR) collections. Plant genetic resources drive innovations and are used to develop new crops, to find new uses for existing crops or to develop improved varieties that produce food, fiber, forage, fuel, ornamentals, medicine or are used for industrial and/or restoration purposes. These collections also provide key sources of genetic diversity for today’s homogeneous agricultural crop production systems. Many crop germplasm collections, including wild relatives, are being evaluated as potential sources of traits for adaptation in cultivated crops. Also, modern biotechnological approaches in areas of genomics have prompted renewed interest in use of germplasm collections for trait discovery. Innovations in basic and applied plant science research at public, private and non-profit organizations nationally and internationally are often spurred by access to these genetic resources.



The Western Regional Plant Introduction Station (WRPIS) of the National Plant Germplasm System (NPGS) is one of four originally established sites in the U.S. dedicated to the conservation and use of plant germplasm vital to agriculture. Established in 1947 in Pullman, WA as a joint undertaking by Washington State University (WSU) and U.S. Department of Agriculture (USDA), the WRPIS has a long history of acquisition, maintenance, characterization, evaluation and distribution of PGR and their associated information. Support for the WRPIS has come from federal appropriations and through the partnership with the Western Regional (W6) Multistate Hatch-funded Research Project. In 2020, the NPGS managed 595,624 accessions corresponding to 15,979 plant species. Within the NPGS, the WRPIS collections hold a significant proportion of the U.S. and world’s germplasm representing cultivated crops and their wild relatives. Primary collections here include cool season food legumes (e.g., pea, lentil faba bean and chickpea), turf and forage cool season grasses (e.g., fescue and bluegrass), temperate forage legumes (e.g., alfalfa and clover), common bean (Phaseolus sp.), horticultural crops (e.g., lettuce, sugar beets, and garlic) and U.S. natives used in restoration along with many other important plant species.



The impact of these diverse collections at WRPIS can be partially demonstrated by the demand for their use. Over the past five years (2015-2019), global stakeholders have requested and received 122,544 distributions in over 3,616 separate requests. Distribution to and utilization of germplasm by researchers and educators in the Western Region in the past five years was for 1,809 (~50% of total) requests and 44,133 (~36%) individual items. These distributions were to public plant breeders and other scientists for research, development and education purposes (e.g., graduate student research) at Land-Grant Universities, other public/private research institutions, USDA sites, non-profit organizations and private industry.


Support for research and service efforts provided by the WPRIS comes from stakeholders. The Plant Breeding Coordinating Committee (PBCC), which represents all U.S. public breeding programs, made the following declaration about access to germplasm in its 2020 renewal document: “Genetic diversity serves as the basis for development of varieties that are critical for providing livelihood resilience and food security. Therefore, it is essential that the U.S. and international communities conserve, characterize, and provide access to diverse germplasm for plant breeding.” Mark Smith, President, North American Alfalfa Improvement Conference (NAAIC), recently stated “Alfalfa is the third most valuable field crop in the U.S. and is a leading agricultural enterprise in many western states. It is a priority of NAAIC to improve performance (yield) and quality of modern-day varieties. Breeding with traits identified in the genetically diverse alfalfa collections of WRPIS is a way to achieve this goal.”. The NAAIC is a community of public/private sector scientists and educators that promotes the exchange of information among researchers involved in alfalfa improvement and utilization around the world.



The proposed project aligns well with national and international agricultural sustainability goals and action plans. These include, among others, the United Nations Food and Agriculture Organizations Sustainable Development Goal 2.5.1 addressing conservation of PGR for food and agriculture and the USDA National Program 301 Action Plan. At the same time, the project is an active collaborative partnership in finding solutions to key issues affecting the Western Region as outlined in the Western Region stakeholder-derived document entitled, “The Western Perspective and the Western Agenda”. In this regard, the WRPIS provides support to the food production, food security and horticultural industries, as well as for conservation and restoration of natural resources in the Western Region.



The importance of the work, and what the consequences are if it is not done.



Plant breeders and other researchers rely on the diversity in agriculturally important crop PGR collections for the identification and sourcing of traits conferring disease and insect resistance, nutritional qualities and yield. As an example, leafhopper insect resistance was identified in alfalfa (Medicago sativa L.) germplasm accessions held at the WRPIS for integration into commercial varieties. A proportion of modern alfalfa varieties carrying this trait has helped reduce its insecticide use, thus benefiting the environment, human health and producers’ economic gain. Alfalfa is the third largest crop (by acreage) in the U.S. and the alfalfa producers of the Western Region states lead in supplying the national and export forage industry markets. Another example of the impact that WRPIS collections have on the U.S. agriculture economy comes from dry pea (Pisum sativum L.), an important vegetable and grain crop with significant production in Western Region states. The nation’s dry pea crop has seen increased production because of its use as an alternate source of plant-based protein concentrates, and consumer benefits come from its high nutritional value (e.g., proteins and phytochemicals). In the last three years distributions for pea germplasm for breeding high protein varieties has increased dramatically to 22,847 packets from the collection of 6,192 accessions. Further, the ‘Hampton’ pea variety released by public breeders in the state of Washington in 2016 has resistance to two important viruses (Pea enation mosaic virus and Bean leafroll virus), both traits sourced from WRPIS germplasm. When viral disease pressure is high, a pea crop cannot be produced without these disease resistance traits. While disease and insect pests limit production, factors affiliated with climate change can also have significant adverse impacts. The WRPIS collections are an essential resource to meet these challenges and are accessed and evaluated for sources of traits/genes associated with abiotic stress response to drought, heat and salinity in order to produce climate-ready crops with improved resiliency.



Many ongoing diverse research projects, a few of which are highlighted below, depend on access to the WPRIS PGR collections. An established project that continues to rely heavily on WRPIS germplasm has been the Cornell University-led NE1710 Multistate Research Project entitled “Improving Forage and Bioenergy Crops for Better Adaptation, Resilience and Flexibility”. This national project has released grass and legume germplasm with improved disease/insect resistance, vigor and other agronomic traits derived from WRPIS germplasm used to broaden the genetic base of breeding programs. Also, WRPIS continues to collaborate on a long-term (>15 years) federal government Inter-Agency project aimed at the conservation and use of native PGR for landscape restoration and improving agroecosystems. This Bureau of Land Management (BLM)-led effort has enabled close to 20,000 native plant collections to become part of the WRPIS and NPGS holdings. These collections and their associated information are cataloged, conserved and available for restoration research efforts, especially targeting western landscapes affected by fires and other natural or human-induced impacts.



Although plant breeders are WRPIS’s primary stakeholders, the need for access to germplasm and its associated information comes from many other areas. Collections of model plant species include Barrel medic, Medicago truncatula Gaertn., one of the first plants with a fully sequenced and published genome. The M. truncatula collection has been useful for research on legume functional genomics and nitrogen fixation. A large collection of purple false brome, Brachypodium distachyon (L.) P.Beauv., T-DNA insertion lines, used in understanding grass species gene function, is also managed by the WRPIS. Discoveries in B. distachyon genetic resources have led to improvement of related food crops like wheat (Triticum aestivum L.) and biofuels crops like switchgrass (Panicum virgatum L.). Over the years, WRPIS germplasm has been requested for a broad range of uses including, but not limited to, agronomy, anthropology, bioremediation, botany, taxonomy, genetics, physiology, entomology, plant pathology, ecology, restoration research, repatriation and for educational and demonstration programs.



Significant effort has gone into assembling and managing plant germplasm collections at the WRPIS. Some of the first plant introductions (PI) held in Pullman date back to the late 1800’s with extensive collection growth since then. Today’s, collections are extremely diverse with 1,133 genera and 4,996 species primarily representing introduced (i.e., non-native) cultivated crop species and many of their wild relatives, vital to U.S. agriculture. Active curation of these collections involves the characterization and evaluation of accessions as well as critical regeneration activities. As collections have expanded over time, so have the capacities for managing information. Comprehensive accession-associated passport, characterization (phenotypic/genotypic) and evaluation data can be freely, publicly accessed with browser-based software via the Germplasm Resources Information Network (GRIN)-Global System (GG) database. Along with germplasm custody, the long-term management of evaluation and genomic data associated with accessions is an important and significant effort, with information access almost as valuable as the germplasm, as it informs selection for research objectives.



If current management of the WRPIS collections were not maintained, future access to its PGR collections will be restricted. Access to the germplasm would be limited due to diminishing quantities and quality of germplasm stock, and regeneration could not be maintained in a timely fashion. Further, any loss of germplasm in the collections most likely would be permanent as it is possible that such germplasm no longer exists where originally sourced. Notably, most WRPIS collections are not native to the U.S. and international access is becoming more restricted. Consequently, loss of access to germplasm could have dire consequences locally, regionally and globally as the need for increased and sustainable agricultural productivity might not be met. PGR have a significant positive impact on economic, environmental and social aspects of our society and its future, and if lost most likely cannot be recreated. Therefore, the collections held by the WRPIS need to be conserved in perpetuity in order to realize their potential for future development.



The technical feasibility of the research.



For over 70 years the WRPIS has been successful in its service and research missions, providing access to PGR and associated information for stakeholders. This achievement comes from fiscal and in-kind support and the collective efforts of both federal and state entities. The WRPIS relies on extensive laboratory, greenhouse, field and office space, much of which is provided as in-kind support from WSU’s College of Agricultural, Human and Natural Resource Sciences (CAHNRS). Approximately 5,000 ft2 (~29,000 ft3) of dedicated cold storage facilities (4oC and -18oC), along with accompanying infrastructure for germplasm distributions, are at the core of the WRPIS service functions. The WRPIS relies on specialized technical equipment and facilities for germinating, threshing and seed cleaning. Research farms and other vital resources that contribute to aspects of the day-to-day operations are also provided at the Central Ferry, WA research farm and the WSU, Irrigated Agriculture Research and Extension Center (IAREC) in Prosser, WA.



The proposed project is attainable because the WRPIS counts on extensive, collective technical and advisory expertise. The WRPIS’s five curatorial programs are led by dedicated scientific and technical personnel with many years of PGR management experience. The station’s research scientists lead programs providing support to curatorial activities including maintenance, characterization, evaluation, genetic improvement and pest management. Administrative and information technology support, along with research farm staff, also play key roles in the WRPIS mission. The WRPIS relies on research, service and clerical support from its Administrative Advisor and their corresponding office. Also providing consultative roles are the Regional Technical Advisory Committee (RTAC) and 10 Crop Germplasm Committees (CGC). The RTAC is comprised of state representatives at Western Region Land-Grant Universities with research programs associated with and dependent on WRPIS genetic resources. Members of the CGCs include NPGS stakeholders with diverse PGR subject matter expertise. Scientific personnel at the WRPIS have a long history and network of productive project collaborations (e.g., WSU) providing valuable input and impact.



Managing critical information associated with NPGS genetic resources is accomplished with GRIN-Global software. This relational database software was developed by the NPGS and international collaborators to specifically manage passport, inventory, taxonomy, characterization, evaluation, distribution data, digital images and other records, genebank workflows, and is continuously enhanced. Characterization and evaluation data collected on WRPIS germplasm can be generated in-house or via collaborations. Many of these accessions-associated datasets are produced by Western Region and other collaborating scientists who provide data back to WRPIS curatorial programs to be curated long-term and made publicly available. GRIN-Global software enhancements by dedicated NPGS software development teams continue to increase system capacity and functionality (i.e., extensibility); the WRPIS helps implement these.



The WRPIS also counts on support from sites like the USDA National Laboratory for Genetic Resources Preservation (NLGRP) in Fort Collins, CO, which holds a safety backup inventory for over 90% of WRPIS PGR. As germplasm enters the WRPIS collections, or as older seed inventories are regenerated, safety backup samples are deposited with NLGPR. The NLGRP also coordinates backup samples at the Svalbard Global Seed Vault, in Longyearbyen, Norway. The National Germplasm Resources Laboratory (NGRL) in Beltsville, MD assists by hosting GRIN-Global data and software development team. The NGRL also coordinates plant germplasm collection and exchanges through its Plant Exchange Office (PEO) and aids in the safe introduction of quarantined germplasm.



The advantages of doing the work as a multistate effort.


The proposed collaborative has a long, successful history of implementation with wide-ranging impacts to the Western Region, the nation and the world. Support for this Multistate Research Project has come from the 73-year partnerships between Washington State University, the Western Association of Agricultural Experiment Stations Directors (WAAESD) and the NPGS. Of benefit to the community is guaranteed access to diverse plant germplasm collections by those with research or education objectives.



Centralized management of genetic resources in one state/location for the region avoids potential duplication of efforts at multiple sites. Although the WRPIS main administrative unit is in Pullman, the ability to conduct regenerations, characterizations and evaluations at multiple sites (i.e., Pullman, Central Ferry, Prosser) means that relevant data is applicable to similar crop growing environments throughout the region. Given its colocation on the WSU campus and proximity to the University of Idaho in Moscow, ID, research collaborations with supportive programs at these institutions are common. Many WRPIS scientists also frequently engage with other regional, national and international efforts to provide data, analyses and research outputs that benefit entire germplasm stakeholder communities. Since this is a regional multistate project, the RTAC plays an important role in advising and providing a channel for communicating individual, regional, and state needs in the context of the WRPIS project management.



Likely impacts from successfully completing the work.



Successfully completing the proposed project will benefit plant science research and educational stakeholder communities by providing continued access to valuable plant genetic diversity and associated information. Optimized protocols for regenerating, as well as for testing and eliminating diseases from germplasm, will help preserve genetic resources long-term and increase availability for safe distribution. The proposed plans include the addition of unique genetic diversity to fill gaps for priority crop species, making this ‘new’ unexplored germplasm available. In-house and external germplasm characterizations and evaluations will provide substantial data that will be linked to accessions in the collection. Access to this data benefits end-users by helping stakeholders refine germplasm request choices. Genotyping PGR collections to study population structure would identify redundancies for consolidation or justification, and potential gaps to target new germplasm acquisition. At the same time, marker associations could precisely identify genic regions associated with or conferring traits and help advance productivity and quality of plant breeding efforts. Past W6 project achievements have clearly demonstrated its ability to provide continued access to, and use of, WRPIS PGR and associated information to stakeholder communities in the Western Region, the nation, and the world (Attachment 1 & 2). These activities emphasize and justify the project’s necessity. Beneficiaries of the proposed service and research include breeders, the plant science and other research communities, and agricultural producers. Furthermore, the general public profits from these efforts by having continued access to safe, nutritious, high-quality agricultural goods.

Related, Current and Previous Work

The WRPIS was established in 1947 as a collaborative effort by WSU and USDA and backed fiscally by federal appropriations and regional Hatch-Multistate funding to support PGR management. A significant proportion (~16%) of all NPGS plant germplasm holdings, important to agriculture and landscape restoration, are conserved by the WRPIS (https://www.ars-grin.gov/). Collections focus on important cultivated crops, their wild relatives and ecologically important native plant species that include cool season food legumes (e.g., pea, lentil, faba bean and chickpea), turf and forage cool season grasses (e.g., fescue and bluegrass), temperate forage legumes (e.g., alfalfa and clover), common beans (Phaseolus spp.), horticultural crops (e.g., lettuce, sugar beets, and garlic), and U.S. natives used in restoration.



Relationships to other national germplasm projects were explored by performing a Current Research Information System (CRIS) search https://cris.nifa.usda.gov/. When using the terms ‘plant genetic resources’ or ‘plant germplasm’ in combination with the word ‘conservation’ 130 and 134 active project were returned. When limiting the search with the same terms to Hatch-funded projects the Boolean search returned 67 and 55 projects, correspondingly. In both search strategies many of the listed projects were, in one way or another, related to or in support of the NPGS plant germplasm collections conservation and use. Searching the CRIS databases for research projects affiliated to crops managed by the WRPIS (e.g., pea germplasm, pea breeding) again led to current collaborations with WRPIS or efforts that rely heavily on access to both germplasm and their associated information. Many of the projects are affiliated with long-standing research programs, predominantly associated with breeding efforts (Byrne et al., 2018), at Western Region Land-Grant Universities (e.g., WSU, University of California, New Mexico State University). In all searches, the three other NPGS collaborative regional multisite projects affiliated with NPGS plant introduction stations (Ames, IA; Geneva, NY; and Griffin, GA) were returned. These sites have similar overall missions, but conserve unique priority crop germplasm, genera and wild relatives assigned to those locations based on their economic impact to the corresponding regions, adaptation, and ability to thrive in those environments (e.g., maize; North Central Region).



Information associated with germplasm accessions in collections is very valuable and must be carefully collected, transferred and curated in order to add value (Weise et al., 2020). The WRPIS makes passport, taxonomic, characterization and evaluation data readily available through https://npgsweb.ars-grin.gov/gringlobal/search, the publicly accessible GRIN-Global interface. The current platform, developed between the USDA Agricultural Research Service, Bioversity International and the Global Crop Diversity Trust, was implemented in late 2015 and is a scalable, robust tool for managing substantial amounts of accession-associated data (Postman, 2010). Data and other information available via GRIN-Global continues to expand with viability tests, regeneration information, CGC-derived (https://www.ars-grin.gov/CGC) descriptor data (Bioversity_International, 2007), and digital voucher images. Accessible, uploaded information also includes large genotyping datasets, mostly associated with cool season food legumes (Cheng et al., 2015; Hellwig et al., 2020b; Holdsworth et al., 2017; Ma et al., 2020) and evaluation datasets for specific crops/genera. Although some capabilities currently exist for hosting genotypic data in GRIN-Global (Volk, 2008), expanded capabilities are being developed to host or to interoperate with other genomic database providers. Additional habitat source information (e.g., phytoregions, geology, and associated species), mostly affiliated with Seeds of Success (SOS) accessions and new collections, is being added as GRIN-Global capabilities are expanded. Much of the data is publicly accessible with network connection and a browser. Germplasm requests are made after a user registers and provides a justification for their need. After a request is vetted, germplasm is provided free of cost for bona fide research and/or educational purposes.



Demand and distributions in the past years have continued to increase for the NPGS (Byrne et al., 2018; Rubenstein et al., 2006) and the WRPIS collections exceeding 40,000 order items (e.g., seed packets/propagules) distributed in 2019 (Attachment 1). Germplasm is distributed to domestic and international requesters. Public, private and non-profit research organizations in Western Region states, which receive thousands of items each year, are some of the primary beneficiaries of germplasm distributions from the NPGS and WRPIS (Attachment 2). Distributing germplasm is a basic WRPIS function and will continue as such into the foreseeable future.



The WPRIS collections have steadily grown since inception. Over the past ten years, ~12,000 accession were added with current holdings approaching 100,000 active accessions that represent almost 5,000 species (Attachment 3). Significant acquisitions of crop wild relatives (CWR) and other U.S. native plant species important to ecological restoration and agricultural production have been included in the WRPIS collections through the BLM-led SOS program with plans for continued collaborations (Barga et al., 2020; Greene et al., 2019). In addition, other germplasm was acquired via targeted collections of reed canary grass (Phalaris arundinacea) from the Mediterranean Basin, and a U.S. native CWR, thicket bean (Phaseolus polystachios). Over 160 pea wild relatives were incorporated from the Global Crop Diversity Trust and Kew Gardens (Dempewolf et al., 2014) and via active Russian (Bogdanova et al., 2018) and Israeli (Hellwig et al., 2020a) collaborations. Other plant exploration and collecting proposal trips have been funded or will be submitted to the NPGS Plant Exchange Office to strategically fill gaps. An exploration trip to Morocco is scheduled for 2022, with other collecting focused on U.S. natives. Much of the germplasm conserved by the WRPIS was introduced to the U.S. (Khoury et al., 2016) and its value continues to increase as germplasm exchanges and international plant exploration and collecting travel have become much more difficult. Complexities in interpretation and application of rules associated with the International Treaty on Plant Genetic Resources for Food and Agriculture (ITPGRFA) http://www.fao.org/plant-treaty/en/, which protects countries’ sovereign rights over their germplasm, will most likely continue to complicate international access (Brink and van Hintum, 2019). Because of limited international access to PGR, WRPIS acquisitions in the future might come from a more domestic focus on U.S. native CWR and restoration species (Dohle S., 2019; Harrison M.L., 2019; Khoury et al., 2013). A review of WRPIS germplasm Crop Vulnerability Statements (CVS) (Grauke, 2016; Volk et al., 2014) is ongoing and will help determine what germplasm is threatened, and to identify and fill collection gaps.



As germplasm is distributed or its viability diminishes over time, it enters a queue for regeneration. In the past five years WRPIS curatorial programs have regenerated more than 10,000 accessions of priority crops and wild relatives using best management practices. Improved regeneration protocols for all crops/genera, especially wild relatives and U.S. natives, continue to be developed for WRPIS managed crops. Protocol improvements have focus on description and management of disease and insect pests and in management of insect pollinators. For example, newly described species of Penicillium pathogenic to Allium sativum (garlic) and other related plant species could impact the already difficult process of regenerating this clonally propagated crop (Dugan et al., 2017; Dugan and Strausbaugh, 2019). However, recent progress was shown for garlic regenerations where bulbils and not basal cloves were used to more effectively prevent infection by Fusarium proliferatum. In alfalfa, gene flow and adventitious presence of genetically engineered (GE) traits (i.e., glyphosate herbicide resistance/low lignin) must be prevented during regenerations. Strategies have been developed to prevent insect transmission of pollen by increasing isolation distances from feral (Greene et al., 2015) or commercial field (Kesoju et al., 2019) GE sources.



Research efforts characterizing and evaluating PGR held by the WRPIS have been extensive.
This work is carried out internally and in wide-ranging collaborations with national and international public, private and non-governmental organizations. More than 300 lettuce accessions were evaluated during germination experiments at different temperatures which identified metabolites associated with tolerance to warmer germination temperatures and potential for selection for heat tolerance (Wei et al., 2020). Research has continued on characterization of adaptation traits and development of empirically derived seed zones for U.S. native plant species predominantly used in ecological restoration. Espeland et al. (2018) and Johnson and Vance-Borland (2016) evaluated adaptation traits and trait plasticity in Poa secunda and Lyemus cinerus, two important grass species used in restoration and range forage activities for which significant WRPIS collections exist. Genetic relationships in lettuce wild relative germplasm were determined using target region amplification polymorphism markers (Beharav et al., 2018). Additionally, genotyping platforms using next-generation sequencing technologies (i.e., genotype-by-sequencing or GBS) (Elshire et al., 2011) or arrays have been used to characterized genetic diversity, relationships and population structure in pea (Cheng et al., 2015; Holdsworth et al., 2017; Ma et al., 2020), lentil (Khazaei et al., 2016); and chickpea (van Oss et al., 2015) germplasm, all WRPIS priority crops.



Marker trait associations have been developed for several important WPRIS crops including a research study that developed codominant SSR and SNP markers useful in breeding Fusarium root rot resistance in pea (Coyne et al., 2015; Coyne et al., 2019). Other research projects have focused on the use of GBS and genome-wide association studies (GWAS) to identify trait-marker associations, or in developing genomic selection (GS) models for improved genotype selection in breeding. An alfalfa program dedicated to enhancing resistance to both biotic and abiotic stressors has developed disease resistant and tolerant germplasm, and identified markers associated with resistance to verticillium wilt (Yu et al., 2020; Yu et al., 2017; Zhang et al., 2014), drought resistance (Yu, 2017; Zhang et al., 2017), salt tolerance (Liu et al., 2019; Liu and Yu, 2017; Medina et al., 2020; Yu et al., 2016) and forage quality (Lin et al., 2020). In addition, several WRPIS cool season food legume germplasm panels have been used to develop genomic resources. These include a sequenced-characterized amplified region (SCAR) marker for the stringless pod trait in pea (Ma et al., 2016), a kompetitive allele-specific PCR (KASP) marker for powdery mildew resistance in pea (Ma et al., 2017), and for identification of quantitative trait loci (QTL) associated to Aphanomyces root rot resistance (Ma et al., 2020), to yield (root/shoot) traits in lentil (Idrissi et al., 2016) and to cold tolerance in chickpea (Mugabe et al., 2019). Several of these projects incorporated innovative approaches including the use of high-through phenotyping in field settings with unmanned aerial vehicles (UAV) and multiple spectral cameras for image capture and analyses (Yang et al., 2020). For example, high throughput phenotyping has been used to identify disease resistant lentil accessions in WRPIS greenhouses and fields (Ma et al., 2020; Marzougui et al., 2019) with ongoing work to estimate alfalfa biomass using high resolution images taken during UAV flights. Compared to manual data collection, these imaging approaches should improve data collection efficiency, accuracy/quality, while reducing phenotyping costs. As this area of research expands, plans are to continue incorporating these technological advancements to successfully characterize and evaluate WPRIS germplasm. Other germplasm enhancement or prebreeding objectives have focused on the selection and development of winter hardy faba beans (Landry et al., 2017; Landry and Hu, 2019; Landry et al., 2015) for use as a rotational cover crop in the Pacific Northwest. Advanced faba bean breeding line populations have been distributed nationally, via Material Transfer Agreements, to university scientists for grain yield trials and to the USDA Natural Resources Conservation Service (NRCS) for cover crop research.



Annual meetings of the RTAC have been held virtually or in person, with on-site venues rotating through different Western Region states with NPGS sites. Every second to third year the meeting is held in person on the WSU campus with visits to the WRPIS, its staff and facilities. These meetings bring together expertise of representatives from 13 Western Region states to discuss project implementation, services, research and stakeholder germplasm and information use. RTAC members – many located on Land-Grant Universities and/or State Agricultural Experimental Stations – are themselves primary stakeholders who have vested interests in the NPGS/WRPIS collections and make use of germplasm accessions in their research (Attavar et al., 2020; Jensen et al., 2020; Ray et al., 2018; Vetch et al., 2020). A select number of NPGS participants representing Western Region and other NPGS sites (e.g., NLGRP) also participate, providing comments and guidance. The state and NPGS site reports are discussed and improved effectiveness and efficiencies are developed and adopted, where feasible, based on stakeholder and RTAC membership feedback.

Objectives

  1. Actively support regional, national and global agriculture and natural resource industries by making edible legume, horticultural, oilseed, turf, forage and native plant germplasm, as well as their associated information, readily available to stakeholders. - USDA and WSU
  2. Optimize regeneration and conservation protocols to provide enough quality and quantity healthy germplasm propagules for distribution and/or for long-term effective conservation. - USDA and WSU
  3. Strategically expand collections by including germplasm with improved horticultural or agronomic traits and/or by filling gaps in collection coverage to enhance conservation of genetic diversity. - USDA and WSU
  4. Generate and provide access to key horticultural or agronomic characterization and evaluation data associated with germplasm collections and/or make available improved germplasm selections. - USDA and WSU
  5. Genotype plant genetic resources to define taxonomic relationships, assess genetic diversity and population structure and/or to perform association analyses with horticultural or agronomic traits of importance. - USDA and WSU
  6. Document plant germplasm collection utilization in the Western Region, nationally and internationally and implement suggested improvements for effective and efficient germplasm management by stakeholder groups. - USDA, WSU, AK, CA, CO, HI, ID, MN, NM, NV, OR, UT, WA, WY

Methods

Dedicated WRPIS personnel, including two Research Scientists (Research Leader/Geneticist [RL] and Research Geneticist [R-Alfalfa]) and five Service Scientists/Curators (Cool-season Legumes, Cool-season Grasses/Safflower, Horticultural Crops, Phaseolus, and Forage Legumes) are supported by both federal (~86%) and multistate Hatch funds (~14%). Federal appropriations cover USDA personnel and their technical staff, with W6 funds supporting six full-time WSU technical staff (Attachment 4). In addition, the WRPIS hires temporary employees (e.g., time-slip student labor) throughout the year, with peaks in summer months, to aid in accomplishing the mission. The USDA-ARS Plant Germplasm Introduction and Testing Research Unit (PGITRU) Research Leader serves as the WRPIS Station Coordinator and oversees project implementation. Five curators, scientists with service and research-oriented responsibilities, lead programs overseeing all PGR management including the day-to-day activities for specific crops/genera (Attachment 5). The curatorial programs include the 1. Agronomy and safflower (AS); 2. Phaseolus (P); 3. Horticultural Crops (HC); 4. Cool-season Food Legumes (CSFL) and 5. Temperate-adapted forage Legumes (TFL). The two Unit Research Scientists focus on investigations in support of effective PGR management or evaluating germplasm for marker development and enhancement. It is also imperative to highlight that much of the service and research proposed is achieved in collaboration with cooperators outside the WRPIS which include other USDA and WSU researchers and many other organizations and individuals. When secured, competitive extramural funding also bolsters WRPIS productivity and allows for the hiring of postdoctoral associates and other technical staff.

Managing diverse germplasm collections is complex (Clark et al., 1997). The WRPIS tries to follow best practices outlined in developed guidelines such as the Genebank Standards for Plant Genetic Resources for Food and Agriculture (Engels, 2003; FAO, 2013). Guidance also comes from the evolving Manuals of Operating Procedures for the NPGS and WRPIS. Most WRPIS germplasm is stored as seed at 4oC and low relative humidity where it can be accessed for distribution. Original seed inventories, or germplasm with low seed quantities queued for regeneration, is stored at -18oC. Germplasm is distributed free of charge, based on justified requester need (research and education), according to developed NPGS distribution policies and congressional mandates. Requests are submitted online through the public GRIN-Global website and individual curators asses requests to determine eligibility. Most distributions are made within a two-week timeframe. In 2017 the U.S. became a full party member to the ITPGRFA with no negative impacts on domestic distributions expected. U.S. policy on use of a Standard Material Transfer Agreement (SMTA) (Brink and van Hintum, 2019) to international distributions is under review.

Management activities, including viability monitoring tests  (Rao et al., 2006; Solberg et al., 2020; Walters et al., 2007), are conducted regularly. As resources permit, specific germplasm collections (e.g., garlic, pea and Phaseolus sp.) are tested for viral diseases that impact plant health and possible distribution. WRPIS curatorial programs will continue to work with NLGRP on effectively backing up germplasm (currently at 90%). Future efforts will focus on optimizing techniques for CWR and U.S. native plant species for which ideal storage (and regeneration) techniques are not available (Walters, 2015) and also longer-term storage conditions (e.g., cryopreservation) for clonally propagated germplasm, that is difficult to back up (Liu et al., 2017; Volk et al., 2004). The WRPIS will continue to back up germplasm with the Svalbard Global Seed Vault through NLGRP.

Accession-associated information will consistently be improved with efforts focusing on passport, characterization and evaluation data content, quantity, and quality. Passport data for newly acquired PGR coming into the WRPIS (e.g., SOS program) includes accurate georeference information with locality data. Descriptor, characterization and evaluation data, along with digital images will continue to be uploaded in GRIN-Global. Possibilities for GRIN-Global hosting larger genotyping datasets (e.g., SNP or sequence data) being generated by WRPIS and collaborators, or for interoperability with databases that host this information continues to be explored. WRPIS personnel will work closely with collaborators to receive, summarize, format and include datasets in GRIN-Global.

At times, specific active collection germplasm is not available for distribution due to low quantities or because viability has declined. In addition, when new germplasm is acquired it is often received in low quantities. For these reasons, germplasm is routinely queued for regeneration (Brown et al., 1997; Clark et al., 1997) (Attachment 6). The WRPIS collections require specific and optimized protocols for efficient regeneration. Regeneration efforts strive to maintain genetic integrity by preventing the following: mixing and mislabeling of accessions, natural/artificial selection, genetic drift, erosion or gene flow in original populations. This is accomplished by meticulous labeling, using described effective population sizes (Johnson et al., 2002), implementing appropriate isolation distances (FAO, 2013), and/or using insect-proof cages to grow self-pollinated species or single accessions for cross-pollinated species.

In most cases, seed is sown in greenhouses early in the year and field transplanted with 100 plants per plot. For cool season grasses, that are highly heterogeneous, and wind pollinated, a 50 m distance between accessions per species is used to prevent gene flow. Approaches that focus on harvesting from an equal number of inflorescences per plant is a practical way to reconstitute original populations (Johnson et al., 2004). Similar measures to prevent genetic mixing are used for several horticultural crops including lettuce and sugar beets, where plants are isolated in cages, by distance or physically in greenhouses. Isolation distances and use of insect proof cages are especially important strategies in preventing gene flow and adventitious presences in alfalfa regenerations. Alfalfa is an outcrossing species pollinated by insects with GE glyphosate resistant commercial production close to regeneration fields. Additional approaches for mitigating potential gene flow and adventitious presence in NPGS collections have been developed in the “ARS procedures and best management practices for genetically engineered traits in plant germplasm and breeding lines v. 73, 14 Feb. 2018”. For alfalfa, sentinel plots are routinely established and subsets of seed lots tested to assess gene flow to field sites and in regenerations (Mezzalama et al., 2010; Tiwari and Randhawa, 2010). Other collections include wild perennial species that may require more than one growing season to mature and produce seed for regeneration. Field regenerations are carried out on one of three research farms with plants most adapted to each of those growing conditions (e.g., cool season legumes/grasses in Pullman).

All bean (Phaseolus sp.) and pea (Pisum sp.) germplasm is regenerated in greenhouses in order to effectively manage insect pests that transmit viruses (e.g., Bean common mosaic virus, Pea seed-borne mosaic virus) that may be seed-transmitted (Nordenstedt et al., 2017; Roberts et al., 2003). Rhubarb and garlic are examples of clonally propagated crops with garlic needing yearly plantings because of its short storage life. For all regenerations, common cultural practices are implemented including appropriate irrigation and fertilization along with insect pest and disease management.

The major acquisition phase for crops managed by the WRPIS are in transition, with fewer accessions being added and strategic acquisitions focusing on CWR (Khoury et al., 2013). In consultation with CGCs and other stakeholders, updated CVS will identify taxa, missing genetic diversity or germplasm with useful traits. All germplasm acquired will follow national and international rules governing PGR including rules of the Convention on Biological Diversity (CBD) and the ITPGRFA. Acquisitions from collecting trips will have standardized FAO multi-crop passport descriptors (MCPD). At least three collection trips are planned. The first has been approved and funded through the PEO for collecting faba bean landraces in Morocco. A second is being planned to focus on U.S. native Phaseolus polystachios in the state of Florida where gaps in coverage of this CWR exist. The third is collection trip planned for a CWR of lupin, the native species Lupinus polyphyllus. The WRPIS looks to continue its long-term SOS collaboration, receiving priority genera and species of U.S. plant species. Donations and exchanges are also possible, where germplasm, advanced lines or varieties have been developed in improvement programs.

As intellectual property rights expire on varieties deposited and held at the NLGRP, they are incorporated into active germplasm collections. Going forward, it will be imperative to monitor these for potential GE traits and to follow protocols and guidance developed in “ARS procedures … breeding lines v. 73, 14 Feb. 2018” in order to maintain genetic purity of publicly held germplasm (Mezzalama et al., 2010; Tiwari and Randhawa, 2010). Accessions of crops with deregulated transgenes (e.g., alfalfa and beet) will need to be managed appropriately.

Most WRPIS germplasm is characterized with standard crop-specific descriptors and digital images (e.g., flower, pods) taken during regenerations. Large-scale, replicated evaluations are conducted in collaborations with extramural funding (e.g., grants) that supplement WRPIS resources. A few planned projects over the next five years are briefly discussed below.

With collaborators at the University of California, a 2,000 plant (200 accession) alfalfa space-plant nursery will be established in Prosser in an effort to generate improved broad-based populations (Simmonds, 1993) that could be used to introgress novel alleles. Replicated space plants will be evaluated over three years for persistence, vigor, and freedom from diseases and insects. At the end of the trial, selected plants will be transplanted to pots for greenhouse crosses. Seed from these synthetic hybrid pools will be released as improved germplasm or planted for a second round of recurrent selection, if funding is secured. Another alfalfa project looks to collect data on alfalfa biomass using high throughput phenotyping in field settings (Yang et al., 2020). An ongoing WSU collaboration uses a UAV and multiple spectral cameras to derive normalized difference vegetation index (NDVI) and other indices. Flights, image captures, and subsequent analyses will be compared to machine and manually collected data for predicting biomass and other important phenotypes.

A multisite (Pullman/Prosser) field trial will be used to develop optimized protocols for germinating, establishing, growing and generating seed for five U.S. native Astragalus species. Little information exists on management of the following species A. canadensis, A. drummondii, A. lentiginosus, A. bisulcatus, and A. lonchocarpus. Alfalfa leaf cutter bees, honeybees and native pollinators use is being evaluated. Plants will be fall-established in a randomized complete block design (RCBD) with three blocks and three treatments of 25 plants per block for each species. Data collected will be similar to that described in Watrous and Cane (2011) focusing on A. filipes, and will include survival rates, phenological and phenotypic measurements, seed quality and yield assessments.

Horticultural crops are characterized every year as they are regenerated, with more comprehensive evaluations coordinated for many years with the sugarbeet (Beta vulgaris) research community. Sugarbeet germplasm will continue to be evaluated (>30 accessions/year) for disease resistance to Cercospora leaf spot, rhizomania, Rhizoctonia root rot, and curly top virus. Additional evaluations examining cyst nematode, root aphid, and root maggot are planned, if funding is secured. Another project will focus on testing optimal DNA barcoding marker(s) (e.g., Rbcl, MatK, ITS-1, ITS-2) (Hebert and Gregory, 2005; Hollingsworth et al., 2016) to genotype lettuce (Lactua spp.) germplasm. Once markers are identified and optimized, they will be used alongside classical taxonomic methods to assign correct identity of lettuce accessions. Plans are to collaborate with USDA Madison, WI on genotyping the garlic collection using GBS.

During routine field and greenhouse regenerations a standard set of digital images, descriptor and trait measurements established by the Pisum and Food Legume Crop Germplasm Committees will be taken. The cool season food legume program will use high throughput phenotyping technologies (e.g. drone-based cameras) to determine biomass and yield estimates in pea and chickpea (Quirós et al., 2019) and heat tolerance in lentil (El haddad et al., 2020). The pea, chickpea and lentil field trails will be planted using a RCBD experimental designs for data collection. Additional evaluations determine seed protein, oil, fiber and starch concentrations for pea and chickpea using near infrared spectrophotometry (NIR) seed scans (Mohammed et al., 2018). Seed mineral nutrient concentrations will be determined for 400 pea accessions established in a RCBD field study (Ma et al., 2017).

During field regenerations, a standard set of digital images and descriptor trait data will be collected on safflower and cool season grass accessions. For cool season gasses, biomass and plant phenology determinations will be made during regeneration using high throughput phenotyping technologies (Galli et al., 2020). A subset of the perennial bluegrass (Poa pratensis L.) collection will be planted in a RCBD to determine turfgrass suitability and biotic/abiotic stress tolerances (Cui et al., 2020).

As DNA sequencing costs have diminished, genotyping entire collections to properly identify germplasm taxonomically, define population structure, develop core subsets, correct mislabeling, rationalize redundancies and to identify useful alleles and germplasm for breeding is becoming more feasible (Belzile et al., 2020; Mascher et al., 2019). Efforts will continue to incorporate genotyping activities for these purposes for priority crops/genera held by the WRPIS.

A USDA/Cornell collaborative effort, Breeding Insight https://www.breedinginsight.org), is developing a array genotyping platform based on the Diversity Array Technology (DArTag) with thousands of single nucleotides polymorphisms (SNP) markers useful in genotyping alfalfa germplasm. Developed markers are currently being validated in alfalfa, and also being tested for their usefulness in other Medicago sp. related taxa. Once an array has been established, and if resources permit, genotyping of ~4,000 alfalfa and wild relative accessions would ensue.

Previous GWAS research identified QTL associated with protein concentration in bean varieties of Andean background (Katuuramu et al., 2018). A similar approach will be taken where GWAS will be utilized on a Middle American dry bean diversity panel (272 accessions) to identify QTL associated with protein concentration utilizing high throughput near infrared phenotyping and the Illumina BARCBean6K_3BeadChip. Plants will be established at two locations (Pullman/Central Ferry) in a RCBD with three blocks and three treatments per block. Phenotype data will be collected over two years with a calibrated near infrared instrument (NIR). For genotypes that are not already part of the BeanCAP diversity panel, DNA will be extracted and genotyped in collaboration with USDA collaborators in Beltsville, MD.

Ongoing collaborations with WSU will continue to dissect/identify genetic factors associated with forage quality and yield in a multisite, multiyear alfalfa evaluation trials. A diverse panel of 200 germplasm accessions and varieties are being measured for 34 quality and yield traits. Plants have been genotyped by GBS and a GWAS approach is being used to associate SNP markers with traits. Initial results found markers associated with fiber, protein and sugar content (Lin et al., 2020) with continued research on gene function. Additionally, research continues to search for marker associations with factors affecting protein degradation.

The pea, lentil and chickpea collections will continue to be genotyped using GBS approaches to determine population structure and marker trait associations. SNP genotypes are available from previous studies for pea and lentil (Holdsworth et al., 2017; Ma et al., 2020) and are being developed for the chickpea core collection for use in the GWAS studies. GWAS experiments will use GapIt 2.0 or BLINK software for the analyses (Huang et al., 2017; Tang et al., 2016). With cooperators, breeding values using genomic prediction models for agronomic traits will be determined for 300 pea accessions (Bandillo et al., 2017; Meuwissen et al., 2001). Also with cooperators reaction to Fusarium root rot in 330 pea accessions caused by Fusarium avenaceum will be assessed and genetic determinants explored with GWAS (Zitnick-Anderson et al., 2018). A newly developed cross legume SNP-chip might also be used to further genotype wild and cultivated pea, lentil, chickpea and lupin PGR. Whole genome sequencing is planned on the pea and lentil GWAS accessions. The GBS approach will also be used on subsets of the cool season grass collections and used for GWAS for biotic or abiotic stress tolerance and agronomic traits (Talukder and Saha, 2017). The safflower collection will also be genotyped (GBS) for GWAS and GS (Talukder and Saha, 2017).

Documenting distributions and their usefulness to requesters is important and provides a way to demonstrate germplasm value and impact. As germplasm is distributed, order numbers and requester information including name, affiliation, address, phone number and email address is logged into GRIN-Global. A recent mandatory field includes now a general use category with optional fields for more comprehensive explanations and instructions. Packaging lists encourage recipients to inspect and provide feedback on germplasm quality, and to invite them to share findings and collection data. Curators can choose to enter such information, providing value to future research efforts. Early in the year a report is generated for distributions from the NPGS/WRPIS to users located in the 13 Western Region states. This report is provided to individual state representatives to contact recipients for feedback. Germplasm recipients respond to questions on condition of the requested material upon arrival and its germination, growth, development and other observations. Additional information is requested on how the material was used (e.g., used in breeding) and outcomes resulting from use of the germplasm, or new varietal releases and/or publications. Responses from germplasm recipients are then compiled by state representatives into a summary report.

The highlights from these state reports, along with activity reports from other Western Region and national NPGS sites that regularly participate in the RTAC, are shared during the annual meeting. Written reports, and corresponding digital copies, are then submitted by individual state representatives and participating NPGS sites to the RTAC and complied into meeting minutes uploaded to the National Information Management and Support System (NIMSS) https://www.nimss.org/ database. Other key participants include Administrative Advisor, USDA administrators from National Programs and Pacific West Area, the Station Coordinator, select WSU/USDA project staff and other relevant NPGS projects in the Western Region/nation. The RTAC assesses the project’s service and research components and its impact, in order to provide recommendations to the WRPIS and administrators on improved efficiencies and to communicate stakeholder requests.

Measurement of Progress and Results

Outputs

  • Continued access to quality/quantity PGR and associated information is the most important of the WRPIS products. Providing free, public access to the large and diverse collections, along with expanded associated information, contributes key support towards innovative agricultural and restoration research that impacts the region, the nation and beyond. Comments: The project described is mostly service oriented with its core mission focused on the long-term conservation and use of agriculturally important PGR. Diverse research activities are also frequently conducted, including incorporation of emerging technologies (e.g., high-throughput genotyping/phenotyping), to effectively meet the project’s core mission.
  • Detailed and optimized protocols for germplasm regeneration, storage (medium/long-term) and conservation are incomplete or don’t exist for CWR and restoration species. Protocols will be developed, expanded, or optimized and made public, to help keep viable germplasm accessible to stakeholders. Comments: Specific outputs: - Detailed information developed and available on management and regeneration activities for U.S. native <i>Astragalus</i> species
  • The WRPIS will improve its PGR collection representation via strategic germplasm acquisition. Increased diversity and access to newly acquired germplasm will provide stakeholders with traits/genes previously unavailable in the collections. Comments: Specific outputs: - Increase diversity and access to germplasm and associated information for: 1) faba bean landraces from Moroccan sources; 2) native bean CWR (<i>P. polystachios/Lupinus polyphyllus</i>); and 3) native restoration/CWRs through the Seeds of Success program
  • Significant subsets of WRPIS germplasm will be characterized and/or evaluated in-house or via external collaborations. Information on highly heritable characters, and significant horticultural, agronomic and/or restoration traits, will be collected and accessible to users. Enhanced germplasm will also be developed and made available. Comments: Specific outputs: - Alfalfa germplasm evaluated with regionally adapted germplasm released - <i>Beta</i> germplasm accessions evaluated for resistance to multiple diseases - High-throughput phenotyping validated for important alfalfa traits - Seed protein concentration data on bean, pea, and chickpea germplasm generated - Large phenotyping datasets generated for cool season food legumes and grasses
  • Taxonomic relationships, genetic diversity, and population structure will be defined for agriculturally important PGR lacking this information. This information will aid in important management decisions including future targets for acquisition, and collection rationalization. WRPIS research programs will develop innovative research techniques to identify genetic markers, that are linked to significant horticultural, agronomic and/or restoration traits, or useful for developing prediction models to benefit plant breeding efficiency. Comments: Specific outputs: - Lettuce accessions genotyped (and phenotyped) for correct taxonomic identity - QTL for protein quality identified in Middle American bean genetic backgrounds - Clonal garlic collections (~300 accessions) genotyped - Marker trait associations validated for alfalfa forage quality - Significant cool season food legume, grass and safflower collections genotyped for marker-trait associations and/or for model predictions/breeding - Genetic architecture of pea seed protein and mineral nutrient concentrations determined using GWAS
  • Validated impacts will be shown for the use of PGR and associated information, as managed and provided by the Western Regional Plant Germplasm Station. Large distributions of germplasm and information will continue to be made to stakeholders in the Western Region. These impacts will be supported by a WRPIS site-specific annual report, as well as a compiled annual report, meeting minutes, and recommendations for the W6 project.

Outcomes or Projected Impacts

  • The impact of access to PGR and their associated information is generally difficult to quantify, especially regarding future benefits. The continued use and increased demand seen in the past few years for PGR conserved and distributed by the WPRIS is one a way to quantify its value.
  • A primary beneficiary of the PGR at the WRPIS has been the region’s and nation’s public and private plant breeding programs. A few examples of how PGR from WRPIS have impacted these breeding programs, producers and ultimately consumers were provided in the justification section of this proposal.
  • Currently, PGR are evaluated for many important horticultural and agronomic traits with specific research on reactions to disease and abiotic stress in alfalfa, cool season food legumes, beans and horticultural crops. Evaluation of U.S. native plant species continues, focusing on characteristics that aid on seed regeneration and those that may play roles in successful establishment for restoration efforts.
  • The collections are also being used in basic research with model plant species (e.g., <i>Medicago truncatula</i> and <i>Brachypodium distachyon</i>) playing important roles in large scale genotyping and sequencing projects that emphasize understanding genetic control of traits and their functions. PGR from WRPIS play important roles in providing plant science research education opportunities and have been used in systematics and taxonomic training. The WPRIS collections are used in so many other unique ways it is impossible to list them all.
  • Increased access to new germplasm added to the collections benefits plant breeding efforts by providing traits/genes not previously available. Well characterized and evaluated germplasm, along with access to these accession-associated traits, allows users to be more efficient by focusing on specific germplasm subsets. Identifying markers linked to traits of interest (e.g., disease, abiotic stressors, yield and/or quality) will enhance breeding efficiency.
  • All enhanced germplasm developed by the WRPIS will become publicly available and can support increased agricultural quality and productivity or benefit restoration efforts. As the world’s population continues to grow and harmful effects of climate become more severe, increased demands will be placed on the agricultural sector for the sustainable production of sufficient quality food and other services. Undoubtedly, continued access to PGR into the future will to play a crucial role in supporting agricultural activities in the Western Region, the nation and the world.

Milestones

(0):Significant annual milestones for WRPIS priority assigned crop/genera include: 1) distribution of thousands of WRPIS assigned germplasm and associated information to national/international requesters; 2) annual accession regenerations (>1,500) for WRPIS assigned germplasm; 3) thousands of seed germination tests monitoring inventory viability; 4) continued collaboration to back up germplasm at the NLGRP and Svalbard; and 5) yearly updating and uploading accession-associated information into GRIN-Global (e.g. inventory, images, evaluation data). Another routine milestone, as the Seeds of Success Interagency collaborations continue, will be to acquire, process and transfer to priority curatorial programs, ~500-1,000 U.S. native plant accession seed inventories annually and upload associated information. The RTAC meeting will be held yearly, with reports on WRPIS status, PGR distribution, use, and impacts along with implementation of suggestions for improvement.

(2022):• Complete first year evaluation protocols for U.S. native <i>Astragalus</i> (SOS) • Establish first year field of regionally adapted alfalfa evaluations space plants (TFL) • If feasible, conduct faba bean plant exploration and collecting trip (Morocco) (CSFL) • Submit bean germplasm exploration and collecting trip proposal (Florida) (P) • Establish multisite Middle American background bean trials (P) • Complete second year phenotyping/GWAS for alfalfa forage quality traits (R-Alfalfa) • Grow/sample known lettuce taxa for DNA barcoding marker optimization (HC) • Complete 4th year pea core seed protein concentration, publish GWAS/GS results (CSFL) • Complete GBS of chickpea single-seed descent/pure-line plant collection (CSFL)

(2023):• Complete second year evaluation protocols for U.S. native <i>Astragalus</i> (SOS) • Collect second year of regionally adapted alfalfa evaluations space plants (TFL) • Crop vulnerability statements updated for the alfalfa collection (TFL) • If funded, conduct bean plant germplasm exploration and collecting trip (Florida) (P) • Plant and collect first year Middle American background bean trials (P) • Complete wet lab protein calibrations for Middle American background bean trials (P) • Genotype Middle American background bean DNA samples, if needed (P) • Identify alfalfa forage quality marker trait associations via phenotyping/GWAS (R-Alfalfa) • Sequence/identify suitable markers for lettuce DNA barcoding (HC) • Complete seed protein concentration evaluations of pea germplasm (CSFL) • Complete/publish whole genome sequencing of pea core collection (CSFL)

(2024):• Compile, summarize and analyze data for U.S. native <i>Astragalus</i> species (SOS) • Collect third year regionally adapted alfalfa, greenhouse transplant selections (TFL) • Produce an updated version of the WRPIS Manual of Operating procedures (RL) • Plant and collect second year Middle American background bean trials (P) • Complete phenotyping Middle American background bean trial (P) • Phenotyping/GWAS alfalfa forage quality study reports and publication (R-Alfalfa) • Identify unknown lettuce accessions with DNA barcoding/classical taxonomy (HC) • Complete/publish whole genome sequencing/GWAS of lentil core subset (CSFL) • Complete pea seed mineral nutrient concentrations, publish GWAS results (CSFL)

(2025):• Report/publish regeneration protocols for U.S. native <i>Astragalus</i> (SOS) • Intermate (greenhouse or field) regionally adapted alfalfa plant selections (TFL) • Report results on Middle American background bean trial (P) • Report DNA barcoding/classical taxonomy results for lettuce accessions (HC) • Complete legume SNP-chip genotyping chickpea subset and GWAS (CSFL) • Complete GBS of cool season grass and safflower collection subsets (AS)

(2026):• Release regionally adapted/improved alfalfa germplasm; begin additional selections (TFL) • Produce an updated version of the WRPIS Manual of Operating procedures (RL) • CVS updated for the trefoil and clover collections (TFL) • GWAS with lupin subset using legume SNP-chip (CSFL) • Complete GBS analysis of additional subsets of cool season grasses and safflower (AS)

Projected Participation

View Appendix E: Participation

Outreach Plan

The WRPIS will continue to participate in outreach activities that promote knowledge and training relating to managing PGR. This will be accomplished in a variety of ways including publishing applied and basic research in scientific peer-reviewed journals that is practical, informative and has impact on management and use of PGR. Information about WRPIS activities will be delivered in the form of scientific presentations (virtual/in person) at professional societies, during invited or guest lecture presentations (e.g., grower groups or class instruction), during workshops, field days, and other outreach opportunities. Other research may be delivered via posters, factsheets and/or flyers. Much characterization, evaluation, and other information associated with WPRIS genetic resources is made available to users via the NPGS GRIN-Global system’s public website (https://npgsweb.ars-grin.gov/), the National Information Management and Support System (NIMSS) https://www.nimss.org/ database, USDA-ARS Unit website (https://www.ars.usda.gov/pacific-west-area/pullman-wa/plant-germplasm-introduction-and-testing-research/), or via other crop-specific websites (e.g., legumes - https://www.legumeinfo.org/; safflower - https://safflower.wsu.edu/). Because of the size and diversity of the plant germplasm collections held by the WRPIS and the varied management activities performed, it has previously hosted several important meetings that included field and facility tours. WRPIS has hosted several Crop Germplasm Committee meetings, the NPGS PGOC and the in-person W6 Regional Multistate project in alternate years.

The WRPIS has also had a long history of hosting students of all levels. Staff believe that playing an active role in training the next generation of PGR managers is critical (Volk et al., 2019; Volk et al., 2019). Visitors for WRPIS tours have included elementary students from local underserved communities to upperclassmen from regional high schools. The large seed storage facilities and diversity in collections are very popular with these young and impressionable minds. High school students from the Native American communities in Nespelem, WA (Colville Indian Reservation) have participated in summer internships as part of the Upward Bound program. The WRPIS also hosts undergraduate students for summer internships from two Hispanic Serving Institutions (Heritage College, Toppenish, WA and Columbia Basin College, Pasco, WA) interested in hands-on training and potential agricultural careers. The WRPIS has also coordinated with the USDA Office of Diversity and Outreach to host several Pathway Interns as well as 1890 Scholars for summer internships and looks forward to continuing these training opportunities.

Because the WRPIS is located on the WSU Pullman campus, there are many student training opportunities. For example, WRPIS staff have had a long history of participating on graduate student (Masters/Ph.D.) research projects and committees. Being on campus also allows for hiring of numerous hourly students throughout the year, especially during summer activities where, many students gain hands-on experience and make decisions to pursue agricultural careers based on their experiences. Over the years, WRPIS has also hosted visiting scientists and many Postdoctoral Associates (WSU and USDA) working collaboratively with staff to advance PGR management science and activities.

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 Regional Technical Advisory Committee (RTAC). The RTAC is comprised of state representatives from the 13 western states and meets annually to assess service and research progress, analyze customer needs and make recommendations to the station. Ex officio members include USDA-ARS National Program 301 Leaders for genetic resources, NPGS personnel at the WRPIS and at other Western Region sites as well as other national NPGS sites that contribute to WRPIS mission (e.g., NLGRP, Fort Collins, CO). Annual meetings rotate through various western sites, with alternate year meetings held at the WRPIS. Currently, Dr. Ian Ray of New Mexico State University serves as the Chair, Dr. Kevin Jensen, USDA Forage Range Research Laboratory, Logan, UT is the Chair-elect, and Dr. Amjad Ahmad University of Hawaii is the Secretary of the W6 RTAC. Dr. Carol Miles of Washington State University was the most recent past Chair. All officers are to be elected for a two-year term and rotate into the next higher responsibility role every two years, providing RTAC continuity.



Administrative guidance is also provided by an assigned Administrative Advisor (currently, Dr. Scot Hulbert, WSU - Associate Dean for Research) and a USDA National Institute for Food and Agriculture (NIFA) Representative (Drs. Ann Marie Thro and Ed Kaleikau, National Program Leaders served in the past). Over the next five years internal benchmarks and accountability systems will be used to assess progress and determine future project needs. In addition to the input from the W6 Technical Advisory Committee, WSU Administration and NIFA, guidance on project management will be sought from USDA ARS National Programs and Pacific West Area office leadership, from the USDA NPGS PGOC, from individual CGCs that represent crops managed by the WRPIS, from germplasm and data recipients/stakeholders, and from feedback and suggestions from external review, as appropriate.

Literature Cited

Attavar, A., L. Tymon, P. Perkins-Veazie and C.A. Miles. 2020. Cucurbitaceae Germplasm Resistance to Verticillium Wilt and Grafting Compatibility with Watermelon. HortScience 55: 141-148. doi:10.21273/hortsci14631-19.



Bandillo, N.B., A.J. Lorenz, G.L. Graef, D. Jarquin, D.L. Hyten, R.L. Nelson, et al. 2017. Genome-wide Association Mapping of Qualitatively Inherited Traits in a Germplasm Collection. Plant Genome-US 10. doi:10.3835/plantgenome2016.06.0054.



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