W4168: Environmental and Genetic Determinants of Seed Quality and Performance

(Multistate Research Project)

Status: Active

W4168: Environmental and Genetic Determinants of Seed Quality and Performance

Duration: 10/01/2019 to 09/30/2024

Administrative Advisor(s):

NIFA Reps:

Non-Technical Summary

Statement of Issues and Justification

The need, as indicated by stakeholders:

Our proposed goals are informed by our close working interaction with our stakeholders who produce and use high quality seeds in the generation of food, feed, biofuel, beverages, natural products, and fiber. They include all individuals and institutions, encompassing scientists in academic-, commercial- and government-sectors, agronomists, horticulturalists and extension agents, who ultimately deliver high-quality seeds and the associated, validated knowledge base to growers. Our stakeholders indicated the need for maintaining or increasing seed quality and asked for new traits that can be incorporated into existing crops. Likewise, they asked for development of new germplasm for novel agricultural endeavors. Our goals for the proposed project focus on developing a broad understanding of how developmental and environmental mechanisms affect seed quality. We then translate knowledge of how to assess and manipulate traits to enhance seed quality (and the criteria with which to measure it) for our stakeholders. An assessment of the existing multistate programs (see below) and the feedback from our stakeholders indicate that we are uniquely positioned to coordinate the necessary research and provide the needed information to our stakeholders. It is clear that a broad, systems-based approach, spanning the basic developmental mechanisms governing seed development, is necessary to understand processes that affect seed quality and how these processes are modified by environmental cues and stresses. We plan to capitalize on our basic knowledge and the use of new technologies (some of which have been generated by our members) to help increase seed quality and performance as demanded by the growers. The long-term goal of our multistate project is to help maintain the high standards and competitiveness of the U.S. seed industry for new and existing crops.

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

U.S. agriculture depends heavily on seed quality to maintain its competitiveness. Our proposed project helps enhance quality and performance of seeds to meet the demands of U.S. agriculture. Poor seed quality causes yield reduction with associated economic consequences of reduced exports, higher food prices, and localized commodity shortages. Although seed quality as a concept is well defined for most familiar agronomic and horticultural crops, it is not one that is defined and applied evenly across all cultivated species. For example, biofuel crops and revegetation programs rely heavily on seeds of high physiological quality that can withstand adverse seedbed conditions, where establishment and survival of the seedlings is the most crucial step. Many of these species have innate dormancy that results in sporadic germination and establishment. Therefore, basic research is needed at the species level to develop the necessary knowledge base for all plant species including wild species, plants already in production or those that are under development for agricultural utilization. This information ensures that growers and producers have the best chance of providing a high quality product to the consumer. This research requires a systematic approach that can only be delivered by a broad and cooperative group of seed- and seedling-scientists. Our proposed multistate project is the only vehicle that can facilitate such research. We know of no other agency or organization that integrates the depth and breadth of expertise to tackle these goals with relevance to the large array of seed crops and species grown in the U.S. 

The technical feasibility of the research:

We have a proven history of carrying out cutting-edge research to develop new knowledge in seed development, seed quality, seed dormancy and germination, and stand establishment. The research proposed in this renewal request focuses on understanding how developmental and environmental mechanisms affect seed quality in various crop species (Objective 1). We will then translate this knowledge using existing and new technology to assess and manipulate traits to enhance seed quality for growers (Objective 2). Our results from genomic, genetic and proteomic approaches associated with Objective 1 will inform some of our practical approaches associated with Objective 2. However, there is already a body of knowledge available to carry out many of the activities associated with Objective 2. Furthermore, the technical feasibility of the research procedures necessary to undertake all activities is standard practice in the case of field-oriented research, or an extension of established principles in plant biology. We do not view these objectives as mutually exclusive, but rather synergistic, representing the continuum between basic and applied research in meeting stakeholder needs. We are one of the longest-running multistate working groups in the USDA, starting as a Regional project in the early 1980’s. Our members are internationally recognized authorities on seed science as demonstrated by our publications, and scientific- and outreach-focused activities described below. We have a proven record of carrying out collaborative research within the present multistate project group and with other cross-disciplinary groups. Given the breadth of expertise and research productivity of the proposed project members (see Attachments), we are confident that we can achieve the proposed objectives fully.

The advantages of doing the work as a multistate effort:

Utilizing a multistate effort by drawing on the expertise of specialized research scientists across many states is the most efficient approach to addressing seed quality on a national level. Although the current multistate project was initiated in the Western Region (1983), the diversity of seed production throughout the U.S. and the lack of any other regional project devoted to seed biology or technology prompted us to expand our research and recruitment efforts to seed biologists at a national level during the recent iterations of the project. In fact, our project has played a critical role in coordinating a diversity of seed biology projects throughout the U.S. Furthermore, due to the comprehensive nature of the objectives, it is essential that we are able to examine seed biology from diverse perspectives, from the molecular to the whole plant level, thereby allowing clarification of the entire biological spectrum and development of specific applications. Therefore, such an approach requires participation of experts at each of these many different levels of focus and with a diversity of contacts for outreach who can help carry out the mission and goals of the project.

An integral aspect of our project is the need to train the next generation of seed scientists and technologists considering the documented decline in the number of seed scientists graduating from land-grant universities in this country. Many of this dwindling number of seed scientists are tasked with educating the next generation at these institutions. Therefore, we face an ever-decreasing number of seed scientists who are trained to educate the next generation (TeKrony, 2006). Seed industries also require continuing university research and the training of seed scientists by land-grant institutions to help them address complex seed physiology problems that impact product development and therefore profitability. The proposed multistate project will continue our current work of integrating the individual activities of our members with all information gained from current state programs across the wide range of species to address problems faced by seed producers and users nationwide. As has been the case in the past, the proposed multistate project will serve as a mechanism to unify seed science research across the U.S. 

What the likely impacts will be from successfully completing the work:

We project the development of solutions that provide an abundant supply of high quality seeds for agriculture as means of maintaining and improving food security for the United States. We will generate new fundamental knowledge about mechanisms underlying seed development, germination, dormancy and storability. We will help increase the seed scientist’s ability to increase seed performance. Other expected outcomes will include increasing the efficiency of food production to preserve environmental quality and further utilization of advanced technologies in seed production. Our project will also contribute to a clearer understanding of how environmental factors affect seed performance in natural and agricultural ecosystems, information which is needed to ensure the continued vitality of native plant populations and the productivity of cropping systems. Finally, our proposed work will provide not only an increased understanding of the factors that influence seed biology, but also practical methods to improve seed performance in the field, both of which are subjects important to the training of the next generation of seed scientists.

Related, Current and Previous Work

Every state and territory in the United States depends on high quality seeds to meet their food, fiber, fuel, beverage, and land management needs; maintain or improve agricultural productivity; and buttress robust domestic and international agricultural trade (TeKrony, 2006).

However, seed quality is a multi-component property linked to fundamental and applied aspects of seed developmental physiology, eco-physiology, environmental interactions, and technological aspects of seed production, delivery and management (Halmer, 2006; Bewley et al., 2013). Simultaneously, despite a crucial reliance on seeds across the nation, most individual states and territories lack public research capacity to produce new knowledge that serves as a foundation for action by stakeholders across seed markets (TeKrony, 2006).

W3168 began in 1983 to address complexities associated with seed quality while coordinating public seed research at the national level. Members in CA and LA have, after many years service, retired, leaving these states without representation. At the same time the membership has recruited new seed biologists in KY, MT, and MI. This is the only multi-state research project dedicated exclusively to developing and disseminating new seed biology information to stakeholders across the US and internationally.

a, and Washington worked collaboratively to organize, promote and deliver the 12th Triennial International Society for Seed Science Conference (Monterey, CA) during September 2017. This conference brought together over 180 delegates from 24 countries to discuss recent advances in seed biology research (https://tinyurl.com/y7cwx4dt). W3168 organizers also collaborated with industry representatives, the Association of Official Seed Analysts, and the American Society for Plant Biology to fund and deliver various components of the conference program including sessions on Student and Post-doctoral Professional Development, industry networking, and panel discussions.

Likewise, our Colorado PI (USDA-ARS) organized and delivered the 2nd Seed Longevity Workshop in conjunction with the International Society for Seed Science during July 2018. This workshop brought together nearly 100 scientists from 22 countries to focus on new findings and future directions related to seed longevity (https://tinyurl.com/yacckzj6). Colorado also organized an oral session and symposia for the C04 Division (Seed Physiology, Production and Technology) at the 2017 International Annual Meeting of the American Society of Agronomy, Crop Science Society of America & Soil Science Society of America.

Furthermore, a recent review of all current multi-state projects listed in the CRIS database (Form 1002, 2019) confirms the distinctiveness of W3168. Four active multi-state projects work with seeds in some capacity: NC-007 (Conservation, Management, Enhancement and Utilization of Plant Genetic Resources); NC1203 (Lipids in Plants: Improving and Developing Sustainability of Crops); NC1205 (Monarch Butterfly Conservation); and NE1838 (Development of a Weed Emergence Model for the Northeastern United States). Yet no significant overlap occurs between objectives or personnel for the proposed project and other existing projects.

For example, NC-007 focuses on collection, maintenance and distribution of germplasm which may include seeds. Seed work in NC1203 centers on enhancing methods to extract lipids from selected crop seeds. NC1205 does not detail seed specific objectives. Rather this project calls for coordinated research on agronomic practices that influence germination of existing pollinator seed mixes. Finally, NE1838 collects existing weed seed emergence data to build a model that assists in weed control efforts.

Related Current and Previous Work of W3168 Participants

Arizona – Extensive research identified key regulatory gene networks that control accumulation of maize (Zea mays) storage proteins and starch during mid to late stages of endosperm development by the Opaque-2 bZIP transcription-factor protein (O2). Collective results provide insights on the complexity of the O2-regulated network and its role in regulation of endosperm cell differentiation and function. A dissection of these functions would enable a deeper understanding of the cell biological processes that underlie endosperm development, which in turn would enable a more targeted engineering of this key seed storage organ for greater seed yield and quality in cereals.

California - Research identified several genes involved in lettuce (Lactuca sativa) seed dormancy (thermoinhibition), including LsNCED4 (ABA), LsERF1 (ethylene), and LsDOG1. The latter was shown to act, at least in part, through association with microRNA 156, and to have additional effects on flowering. A different project focused on measuring single-seed respiration during germination and demonstration that respiration rates were closely correlated with seed vigor. Investigators applied threshold models to seed populations then extended model use to cell biology and ecology. They also developed the “Dry Chain” concept for preservation of seeds and commodities, particularly in humid climates.

This work is important to continue because identification of causal genes underlying lettuce seed dormancy enables use of gene editing and breeding to reduce dormancy and improve germination percentage and uniformity in regions where this commodity is grown for US consumption. Controlling flowering could enable rapid cycling for breeding and improve nicking for pollination in hybrid seed production. Automated seed respiration measurements could replace costly and labor-intensive vigor tests. Modeling can be utilized with digital imaging to identify seed subpopulations and guide seed separation and conditioning for upgrading seed quality. Drying and packaging are critical for preserving seed quality in humid environments. The Dry Chain is being implemented in Bangladesh, India, Thailand and other countries. Opportunities exist to apply Dry Chain to humid seed producing regions of the US.

Florida - Natural resource managers, conservation organizations and native plant producers know which plants they want to utilize but cite a gap in seed biology information as a primary challenge to their operations. A collaborative project: 1) identified key wild species in high demand but lacking essential seed biology information and 2) transferred new information regarding the germination ecology, abiotic stress tolerance and variation of seed quality for several, stakeholder-selected species. Stakeholders can use this information to better plan production, land restoration and conservation efforts. Continuation of this research can enhance seed production within the specialized U.S. wildflower seed industry.

Iowa - Abiotic and biotic stresses during vegetative and reproductive stages of plant development can detrimentally affect seed quality and composition. These changes can directly affect corn and soybean (Glycine max) market value and farmer’s revenue.  The use of organic- and chemical-seed treatments and their impact on seed quality are of vital importance to farmers and stakeholders.

Seed researchers worked with the National Seed Health System (NSHS) to develop and improve seed health testing methods that benefit the seed industry and those involved in producing and exporting seed at the global level. Researchers also developed risk assessment models for seed-transmitted pathogens with potato spindle tuber viroid (PSTVd) and tomato canker as model systems. A universal framework for a model that incorporates quality management practices into risk assessment has been developed. Continuation of this work benefits society by enhancing seed health systems that improve seed quality thereby improving seed stakeholder livelihoods and securing national food supplies.

Kentucky - The maintenance of seed quality, defined as storage longevity and establishment vigor, is dependent on the seed’s capacity to withstand desiccation and to remain viable and vigorous while desiccated. Investigations focused on the involvement of intrinsically disordered proteins (LATE EMBRYOGENESIS ABUNDANT Proteins: LEAPs) and raffinose family oligosaccharides (RFOs) in the capacity to withstand desiccation. At the same time, research elucidated how PHYTOCHROME INTERACTING FACTOR1, which represses the completion of germination, rapidly degrades following exposure to light. This work yielded models on genetic control of seed vigor in maize and how germination is either hastened or prolonged based on cellular capacity to sense light of specific spectral quality. Future work in this area can elucidate key factors in maintenance of seed quality for extended periods and environmental regulation of germination.

Investigators also determined the morpho-physiological aspects of water gap in those families where the water gap has not been adequately described. Physical dormancy (PY) is caused by one or more water-impermeable layers of palisade cells in the seed (or fruit) coat.  One monocot and 16 eudicot families contain species that have PY. Seeds with PY cannot imbibe water even under favourable environmental conditions. Specialized structures are involved in occlusion of the water gaps. The breaking of PY involves disruption or dislodgement of ‘water gap’ structures, which act as environmental ‘signal detectors’ for germination. Ten water gaps responsible for maintaining physical dormancy were newly described and a new scheme for categorizing and defining water gap complexes was developed for the known species with physical dormancy. Identification of these diverse water gap structures and the environmental cues that break PY have significantly enhanced our understanding of germination in physical dormant seeds. This information will prove to be useful for stakeholders involved with commercial seed production and ecological restoration.

New York - Seed treatments are routinely applied worldwide for efficient pest management. A chemical must be able to reach the embryo to be efficacious at eradicating internal seed-borne pathogens. Therefore, a series of fluorescent piperonyl amides were obtained and a novel combinatorial pharmacokinetic technique was developed that provided a range of compounds that could reach maize and soybean embryos. Furthermore, broccoli (Brassica oleracea) seed coating formulations were developed using soy flour as a sustainable bio-stimulant. Broccoli seedling root and shoot growth in the lab showed significant improvement from bio-stimulant seed coating treatments compared with controls. Further research in this area would yield new seed coatings for various crops seeds.

Oregon - Research was conducted in two main areas that affect seed quality: 1) evaluate seed deterioration of some crops in storage and suggest ways to reduce the extent of deterioration and 2) develop a technique to distinguish various species of noxious weed seeds within a seed lot such as amaranth (pigweed; Amaranthus palmeri), which grows quickly and aggressively causing significant yield loss in many crops. This work identified the magnitude of seed deterioration over time associated with factors including: seed anatomy, chemical composition, storage conditions, initial quality, insects, rodents and micro-organisms. Likewise, a new method to identify Palmer amaranth based on morphological characteristics. More work in this area can contribute to creation of storage standards to maintain seed quality and methods to improve quality at the seed lot level.

South Dakota – Seed dormancy (SD) is an adaptive trait of both ecological and agricultural importance. Previous research identified a set of quantitative trait loci (QTL) for seed dormancy (qSD) in the conspecific weedy (Oryza longistaminata and O. punctata) and cultivated rice (O. sativa). This work identified gene networks involved in endosperm-imposed dormancy, endosperm development, hormonally controlled germination capacity and imposition of embryo dormancy. These accomplishments provided mechanistic insights into the development and evolution of SD and new genes with which to manipulate germination ability in crop breeding.

Texas - Vegetable seedlings planted in southern U.S generally experience simultaneous high air temperatures and drought stress episodes causing transplant shock and reduced stand establishment. Research focused on screening and optimizing exogenous plant growth regulators (PGRs), bio-stimulants, and nitrogen rates to increase the quality and ability of seedlings to mitigate abiotic stresses. Ethylene applied to onion seeds promoted fine root development and root surface area in seedlings, while hydro-priming proved to be a simple and viable method to enhance onion (Allium cepa) germination and seedling root traits. Understanding growth and physiology in response to transplant manipulations can provide the basis for developing practical solutions for nurseries and farmers to increase the quality of the transplants and survival when established in the field under stressful environments.

Virginia – A project used ATR-FTIR (Attenuated total reflectance - Fourier transform infrared) spectroscopy to rapidly and non-destructively detect BFB (bacterial fruit blotch; Acidovorax citrulli) infected seeds. Preliminary results show that ATR-FTIR chromatograms in the mid-infrared (MIR) region hold potential to rapidly and non-destructively detect single-seeds infected with 10-5 colony forming units of A. citrulli based on differences in ester C-O (1180-1000 cm-1) that only occurred in infected seeds. A second project developed a new type of thermogradient table to more realistically analyse seedling emergence in the lab. A final project identified seed morphological characteristics that regulate sweet basil (Ocimum basilicum L.) germination. Continued research in these areas can provide new technologies to enhance seed health and vigour assessments.


  1. 1. Understand how developmental and environmental mechanisms affect seed quality
  2. 2. Capitalize on new technologies to assess and manipulate traits to enhance seed quality


Objective 1: Understand how developmental and environmental mechanisms affect seed quality

Arizona, Kentucky, Montana (new member) and South Dakota will use Arabidopsis (Arabidopsis thaliana), maize, rice, and wheat (Triticum aestivum) as model crop species to expand knowledge of embryo and endosperm development with specific reference to gene networks controlling cell division, cellular differentiation, accumulation of dry matter reserves implicated in maintenance of seed quality, and expression of pre-harvest sprouting. Researchers will employ an array of biochemical, molecular biological and field-based approaches. For example, Montana and South Dakota will combine greenhouse and field plantings of transformed crops to assess vulnerability of high grain protein content (HGPC) genes (GPC-B1) to late maturity alpha-amylase activity and pre-harvest sprouting using at least 12 pairs of HGPC near-isogenic lines with confirmed absence or presence of the GPC-B1 gene. South Dakota’s results related to basic HELIX-LOOP-HELIX (bHLH) transcription factors regulating down-stream gene targets that control embryo dormancy and thus pre-harvest sprouting can inform Montana’s expected outcome of identifying lines susceptible or resistant to pre-harvest sprouting.

Likewise, molecular work including RNA-seq, Y1H, BiFC, Co-IP, ChIP-sequencing and phage display conducted in Arizona, Kentucky and South Dakota is expected to clarify the roles of certain protein orthologs and transcription factors related to embryo and endosperm development. Furthermore, work in Kentucky on identification of raffinose family oligosaccharide intermediaries and their influence on plant responses to stress using transposon-tagged maize and Arabidopsis mutants and over-expressing lines can inform work in South Dakota on QTL identification, genotype × environment studies and gene effect evaluations for control of seed longevity of weedy species in a soil seed bank.

Florida will examine the impacts of simulated climate change on expression of ecotypic differences related to seed functional traits for widely-distributed coastal dune species. The goal is to disentangle the influence of increasing thermal and aging stress on the germination and quality of seeds collected along a continental-scale latitudinal gradient. Florida will use Sea oats (Uniola paniculata L.) as a model system to assess germination capacity from 12 seed lots collected across the Gulf (southeastern Texas to southwestern Florida) and Atlantic (Miami, FL to Cape May, NJ) coasts using a reciprocal sowing germination chamber experiment. Chamber temperatures follow seasonal temperatures for different regions along the coasts. Subsequently, researchers will assess vigor differences from seed lots across the latitudinal gradient via controlled deterioration tests. Finally, the project will comparatively assess the capacity for production of physiological defense mechanisms (i.e. detoxifying enzymes and antioxidant compounds) from selected lots exposed to aging stress according to standard spectrophotometric methods. Potential collaboration exists between Iowa, Kentucky and Oregon related to assessing stress responses for this new project. Florida will also organize an oral session, poster session and symposium for the C04 Division (Seed Physiology, Production and Technology) at the 2019 International Annual Meeting of the American Society of Agronomy, Crop Science Society of America & Soil Science Society of America.

Research at Iowa will focus on investigating the fundamental mechanisms for seed quality changes produced by environmental stress during on-plant seed development. Plants will be grown in growth chambers, greenhouses and field environments allowing for artificial or natural stress. Seed quality assessments (e.g. germination percent, rate, uniformity, seedling emergence) will take place to gain a better understanding of variation in seed quality following exposure to various stressors. Seed quality assessment occurs at New York, Florida, and Oregon. These stations represent potential locations for collaboration.

Objective 2: Capitalize on new technologies to assess and manipulate traits to enhance seed quality

Florida recently discovered that differences in intra-specific seed mass within lots of mature seeds plays a role in germination response following thermal and aging stress. This implies that seeds of different mass may deteriorate at different rates. The current research goal for this new project is to provide a physiological basis for the differential germination responses observed in seeds of different mass following exposure to abiotic stress specifically high temperatures and aging. Florida will assess respiratory metabolism (O2 uptake) on an individual seed basis using a next-generation VIM Technology respirometer. This equipment can be shared with other members. For example, Florida and Virginia can collaborate given previous seed respiratory research conducted in Virginia.

Kentucky will undertake a new line of research focused on determining the basic germination parameters for industrial hemp (Cannabis sativa). This will include information related to temperature and the development of a seed priming protocol. Hemp is a relatively new crop for U.S. and understanding the relationship between soil temperature and germination in untreated and primed seeds will impact grower’s decision making relative to planting date, weed control, as well as optimal flower and seed set. The cardinal temperatures for industrial hemp will be determined for several accessions utilizing a thermogradient table. A thermal time model will be developed from the generated data. Once this information is available, osmotic- and solid matrix-seed priming protocols will be developed utilizing two of the seed accessions. The impact of seed priming on germination and stand establishment will be evaluated for seeds germinated over a range of non-optimal conditions. New York, Texas and Virginia indicated collaboration interests to research industrial hemp.

Iowa plans to identify seed vigor QTL regions to facilitate breeding for enhanced seed quality in soybean. Researchers will evaluate soybean seed composition and seed vigor QTLs in Sudden Death Syndrome- (SDS-; Orbivirus spp.) and soybean cyst nematode- (SCN; Heterodera glycines)-tolerant and susceptible soybean varieties grown under disease pressure. Concurrently, Iowa will optimize RNA extraction and purification protocols for seed transmitted virus and viroids of multiple vegetable and crop seeds. This work complements research in New York, Oregon and Virginia.

New York, Oregon and Texas will focus on seed treatments and coating technologies for efficient pest management, delivery of bio-stimulants and compounds to ameliorate environmental stress during planting. New York will adapt rotary pan coating for seed treatment application of organic and chemical seed treatments for the control damping-off caused by Pythium spp. Laboratory bioassay and field studies will be conducted to assess efficacy of treatments in comparison with the non-treated control. Oregon and Texas plan to examine the role of various plant growth regulators to manipulate seed maturity, harvesting efficiency and seedling transplant survivability. Field trials will be conducted to analyze the survivability of seedlings at harvest and after subsequent transplant into the final crop spacing. Untreated plots, as well as PGR’s would be applied at different rates. Seed yield, dry weight, moisture and chlorophyll contents, and yield would be assessed. Seed quality at different seed development stages and upon maturation would be evaluated to determine the effects of PGR on viability and vigor. Moreover, seedlings of high value vegetables such as onion, artichoke (Cynara cardunculus), tomato (Solanum lycopersicum), pepper (Capsicum annuum), lettuce and watermelon (Citrullus lanatus) will be assessed for root-shoot growth traits in response to media bio-regulators pre- and post-exposure to heat and drought stresses. Replicated trials will be conducted under different environments. Image analysis will be performed to characterize specific root traits (WinRHIZO v.5) and green-cover area (ImageJ 1.51s).

Kentucky and Virginia will develop and optimize non-destructive seed analysis procedures using near-infrared spectroscopy (NIRS) and ATR-FTIR (Attenuated total reflectance - Fourier transform infrared) spectroscopy to evaluate seed quality. NIRS system analysis offers the potential for non-destructive evaluation of several seed quality attributes including seed vigor. Eighty seed lots of a single soybean cultivar will be assessed for standard seed germination and vigor using accelerated aging. Seed lots with greater than 85% standard germination and a range of seed vigor attributes will be used to develop models for seed germination and vigor using NIRS. Once the models are developed, they will be evaluated for goodness-of-fit using blind test samples.

ATR-FTIR is crucial for providing a non-destructive procedure for detecting diseased and dead seeds and removing them from commercial seed lots. An alternative approach will be to use gusseted thermogradient tables with soil, LabField technology, for laboratory germination testing to more accurately predict field emergence. Lots of cucurbit seeds that are disease free, infected, and dead will be assessed using AOSA standard laboratory seed germination tests, outdoor field emergence tests, and LabField tested in soils across a range of temperatures in a laboratory.  Subsamples of the same seed lots will be analyzed using ATR-FTIR. Germination test results will be modeled and compared for their ability to predict field emergence and to detect diseased seeds. Concurrently, ATR-FTIR scans and LabField simultaneous germination testing across a range of temperatures in soil will be compared to field emergence using cucurbit seeds and other species of vegetable crops to see how well the new technologies can predict field emergence.

These new projects derive from research collaboration conducted between Iowa, New York, Oregon, Texas, and South Dakota. For example, South Dakota plans to collaborate with Kentucky to evaluate individual and combined effects of seed dormancy genes (SD12a, b and c) on resistance to pre-harvest sprouting (PHS) using NIRS.

Michigan (new member) will determine specialty crop seed producer needs pertaining to seed quality management and assessment, determine appropriate quality standards for the identified crop, and identify and test suitable technologies to improve seed producer assessment and management to assist in consistently grading to these seed quality standards. Small scale organic- and specialty-crop growers will be identified and interviewed to determine specific practices and needs to enhance the quality of seeds they produce. Germination and vigor standards, identified by these seed producers in collaboration with the PI, will be determined for specialty crops. Multiple technologies to streamline small seed producer management and assessment of seed quality will be evaluated by the growers. The seed quality standards and the means of assessing seeds based on them, are areas of overlap and collaboration with Florida, Kentucky, Iowa, and Oregon.

Measurement of Progress and Results


  • Objective 1: Understand how developmental and environmental mechanisms affect seed quality Comments: Subject Area: Germination ecology and climate change. Collaborating partners: FL, IA. • Assess potential climate change impacts on germination ecology of widely-distributed species in SE North America, including coastal buffers. • Better understand how environmental stress impacts seed quality characteristics. • Collection strategies will be revised for land management practitioners, including those managing coastal regions. Subject Area: Developmental and environmental mechanisms affecting seed quality. Collaborating partners: AZ, KY, MT, OR, SD, WA. • Demonstration that raffinose family oligosaccharides (RFOs) pathway intermediates cryptically influence maize germination and longevity. • Identification of LATE EMBRYOGENESIS ABUNDANT PROTEIN (LEAP) interacting proteins in several plants. • Elucidation of gene regulatory network(s) associated with seed storage proteins and starch at various stages of corn seed development. • Development of mutant maize lines in transcription factor genes downstream of OPAQUE-2 and with novel defects in endosperm development and kernel quality. • Identification of wheat lines susceptible or resistant to late maturity alpha-amylase or preharvest sprouting (PHS). • Acquire phenotypic data for wheat lines resistant or susceptible to late maturity alpha-amylase or PHS. • Evaluate plant growth regulator treatment impacts on seed composition and yield. • Identify cereal proteins capable of interacting with SEED DORMANCY12 (SD12a or c) to form heterodimers to regulate gene expression. • Identify SD12a and/or c downstream genes in regulatory networks enhancing embryo dormancy. • Develop isogenic lines for SD12a, b and c, and their combinations. • Obtain information about seed dormancy gene main (additive) and epistatic effects on cereal PHS resistance.
  • Objective 2: Capitalize on new technologies to assess and manipulate traits to enhance seed quality Comments: Subject Area: Seed technology and quality assessment. Collaborating partners: FL, KY, CO, NY, IA, OR, SD, and VA. • Evaluate across-lab uniformity and reliability of U.S. seed health testing for high-profile pathogens. • Make available QTLs for disease resistance and seed vigor to researchers and plant breeders enhancing seed quality. • Develop non-destructive (ND) NIRS models in soybean that can predict seed lot germination potential and seed vigor levels on a single- or batch-seed basis. • Develop ND NIRS models to predict resistance to PHS in cereals. • Develop ND ATR-FTIR models to assess seed quality. • Obtain insight into the role of seed mass on abiotic stress tolerance and maintenance of seed quality. • Developed improved seed storage guidelines for wild flower accessions. • Evaluate LabField thermogradient testing in soil across a range of temperatures to predict field emergence. Subject Area: Seed and transplant technology for improved stand establishment in conventional and specialty crops. Collaborating partners: NY, IA, KY, TX, VA. • Determine seed priming protocols for hemp assessing their ability to complete germination at non-optimal temperatures. • Optimize fungicidal hemp seed treatments for control of damping-off. • Develop pre-transplant conditioning methods to improve vegetable transplant quality. • Evaluate wild-type Solanum spp. as effective rootstocks to enhance growth and yield under heat- and water-deficit conditions. • Determine the physiological-biochemical basis for enhanced growth and yield under heat- and water-deficit conditions conferred to grafted tomato from wild Solanum species. • Establish cardinal temperatures for germination of hemp cultivar seed lots.

Outcomes or Projected Impacts

  • • A collaborative, inclusive, nationwide seed biology research group, poised to participate in opportunities for research and funding.
  • • The education capacity of this nation’s public universities in all aspects of seed biology to the next generation of seed biologists will be safeguarded through this inclusive, unifying collaborative assemblage of diverse experts.
  • • An increased understanding of the potential and limitations of non-destructive (ND) seed sampling on an individual seed basis for: live-dead seed sorting; vigor assessment and; individual seed compositional traits linked to human and livestock nutritional requirements and stakeholder-defined end uses.
  • • ND techniques, once established and rigorously tested on diverse seed species will be translated to our stakeholder groups to facilitate their production of high quality seed for their clientele, improving stakeholder profitability and end-user (farmer) satisfaction.
  • • Production and testing of a state-of-the-art seed vigor and seedling emergence testing platform (LabField thermogradient analysis system). This device will be sold to various academic scientists and commercial seed producers which will greatly aid the state-to-state uniformity of seed and seedling vigor assessments.
  • • New knowledge will be acquired of seed metabolism and biology for improvement of all aspects of seed, from its capacity to store well, to the resulting seedling vigor from high quality seed. From the nutritional quality of seeds for humans and livestock, to its capacity to withstand sub-optimal seedbeds due to biotic or abiotic detractors. This new knowledge will be shared with academics and other stakeholders through journal articles, web pages, and national and international conferences
  • • The potential and limitations for favorably manipulating extant seedlots and seedlot production through the use of cutting edge seed coating treatments, priming regimens, hormonal applications, grafting techniques, and harvesting schedules will be assessed.
  • • The timing of seed harvest and its association with performance in subsequent seed storage over a variety of conditions for wild accessions will be unveiled.


(1):Successful coordination with the NRSP1: Multistate Research Information Management and Impact Communications Program administration for a W4168-directed webinar to better educate faculty and staff on how to write impact statements. This has been identified by the NRSP1 as a persistent shortcoming when retrieving information from working groups. The NRSP1 has, as one of their identified activities, the provision of education in this vein to interested parties. Betterment of our capacity to provide NRSP1 information will assist us, through them, to communicate impacts from multistate research and Extension activities to decision-makers and stakeholders alike. This will allow us to do justice to our advancements in important issues in seed biology and the potential they hold for improved food production and security.

(2):Coordinated proof-of-concept that the LabField thermogradient analysis system accurately predicts field performance for a variety of crops. Collaborating members include FL, KY, NY, IA, OR, SD, and VA. The system will be made commercially available to interested stakeholders (industry and academic). The advent of the LabField with protocol for non-destructive seed testing (ATR-FTIR, NIRS) for a variety of valuable traits during the first half of the project will permit rapid testing of the efficacy of seed treatments and genetic manipulation of protective programs at later stages of the project (see Year 4 below). It is anticipated that breakthroughs in ND seed evaluation for dormancy traits and the genetic proclivity for PHS, will greatly benefit genetic programs evaluating germplasm varying in these traits. In particular, the capacity to nondestructively assess hemp seed quality before dissemination to farmers will prove a boon to this nascent agricultural industry

(3):Information on the impact of the raffinose family oligosaccharides capacity to protect seeds and vegetative tissues from abiotic stress will be made available via collaboration led by KY to the scientific community with which better-informed decisions can be incorporated into breeding efforts currently underway to reduce/eliminate these important protective and pre-biotic sugars from foodstuffs. The surging hemp industry will be provided with recommendations pertaining to planting date based on optimal germination temperatures spanning multiple states, via work between KY, NY and VA in which this commodity is increasingly planted.

(4):A comprehensive methodological advance in ND imaging of seeds to permit single-seed assessment of viability, vigor, and componential quality and quantity will be made available. In addition to assisting industrial stakeholders with assessments of their seed lots, the academics within this project will benefit from these methodologies to evaluate the efficacy of seed treatments (seed priming and the application of plant growth regulators) to protect and enhance seed vigor. The capacity of germplasm with enhanced/reduced: susceptibilities to disease resistance; seed mass; main and epistatic effects on cereal PHS resistance; alterations in endosperm development and seed quality and; late maturity alpha-amylase, to maintain high vigor and produce an excellent crop will be accurately and more rapidly and robustly assessed with this technology. Members from AZ, FL, KY, MT, NY, IA, OR, SD, and VA contribute here. Greater insights into how environmental stress during development impacts subsequent seed quality will be obtained. These insights will range from the deployment, efficacy, and molecular function of established endogenous protective molecules (e.g. LEAs and RFOs); include nutritional aspects (seed storage proteins and starch as well as overarching gene regulatory network(s) orchestrating their production); to general (QTL) and specific (SEED DORMANCY12, OPAQUE2, and late maturity alpha-amylase production) gene products underlying cereal seed quality and PHS resistance. AZ, KY, MT, and SD address this portion. A greater sophistication in our understanding of how climate change might alter diverse species’ seed viability and vigor in different habitats will be acquired and disseminated to stakeholders. Emphasis will be on species vital to the maintenance or re-establishment of crucial, sensitive areas (e.g. wetlands facing developmental-; exotic species competitive-; or habitat erosion-pressures; Gulf and East coast storm barriers). FL and IA work collectively towards this aspect.

(5):Further erosion of the capacity to educate individuals in aspects of seed biology at public universities in this nation will be stemmed by all participants. How and if LabField, non-destructive NIRS and ATR-FTIR testing, can assist in establishing both the across-lab uniformity and reliability of U.S. seed health testing will be documented. These systems can be tested to determine how robust novel QTLs for disease resistance, seed vigor, and resistance to PHS, are. KY, MT, NY, IA, OR, SD, and VA collaborate in this area.

Projected Participation

View Appendix E: Participation

Outreach Plan

The members of W4168 comprise a group of highly dedicated seed biologists who excel in the communication of their research findings. All members of the W4168 project are active participants in seed research at universities and federal facilities throughout the country. They provide leadership in this vital area through undergraduate and graduate instruction, as well as by mentoring graduate and undergraduate research. A number of our members conduct extension workshops to provide the seed industry with a thorough orientation to seed biology fundamentals, as well as the latest cutting edge results. For example, the Seed Biotechnology Center at UC Davis offers courses in seed biology and breeding technologies to the public and seed professionals that incorporate the latest information generated through W4168 (http://sbc.ucdavis.edu). The Iowa State Seed Center also offers regular courses and workshops in topics related to seed biology, conditioning and marketing (http://www.seeds.iastate.edu/). VA and CA will write a new textbook on Vegetable Seed Biology, Production and Quality. Moreover, VA has been a leader in utilizing distance education to deliver seed courses to national- and international-student groups.

As documented in the annual reports of the previous project (W3168), W4168 members regularly publish their finding in top-tier, peer-reviewed journals, targeting both the general plant biology and seed biology communities. W4168 members are also active participants and presenters at various professional society annual national/regional meetings, as well as at the major workshops and symposia sponsored by the International Seed Science Society and the International Society for Horticultural Sciences. W4168 members serve on journal editorial boards and/or as ad-hoc manuscript reviewers, publish books and book sections on seed biology (Allen et al. 2007; Bewley et al. 2012; Bradford and Nonogaki 2007; Perry and Yuan 2011; Pluskota et al. 2011), and obtain patents for intellectual property (Frey et al. 2009; Madsen et al. 2010; Taylor et al. 2011).

W4168 members also interact with seed industry groups on a regular basis. For example, FL works with native seed industry members to develop collaborative research projects that enhance the emerging seed production industry in this region. In California, an organization called Seed Central enhances communication and partnerships between UC Davis and the surrounding seed industry (www.seedcentral.org). Seed Central is a collaboration between the Seed Biotechnology Center (directed by W4168 member CA) and SeedQuest.com (the premier website of the global seed industry). This organization sponsors monthly networking events and facilitates collaborative public/private research related to seeds and crop improvement. It also continues an annual Vegetable Research and Development Forum to discuss research needs and common issues in the vegetable seed industry, at which W4168 members from NY and CA presented their research programs.


Organization will follow recommendations for the Standard Governance for multistate research activities including the election of a Chair, a Chair-elect, and a Secretary. All officers are to be elected for three year terms, as follows: a Secretary will be elected annually, then become Chair-elect in the second year, and Chair in the third year. Administrative guidance will be provided by an assigned Administrative Advisor and a NIFA Representative. The W4168 welcomes and encourages participation of expert seed biologists affiliated with State Agricultural Experiment Stations, the Agricultural Research Service, and colleges or universities, as is consistent with the Multistate Research Fund mission of the Agricultural Research, Extension, and Education Reform Act of 1998.

Literature Cited

Allen P.S., Benech-Arnold R.L., Batlla D., and Bradford K.J. 2007. Modeling of seed dormancy. In: Bradford KJ, Nonogaki H (eds) Seed Development, Dormancy and Germination. Blackwell Publishing, Oxford, UK, pp 72-112

Bewley J.D., Bradford K.J., Hilhorst H.W.M., and Nongaki H. 2013. Seeds: Physiology of Development, Germination and Dormancy. Springer.

Bradford K.J. and Nonogaki H. (eds) 2007. Seed development, dormancy, and germination. Wiley-Blackwell, Oxford, U.K.

“Form 1002: Active MRF Support Projects, Coordinating Committees/Education or Extension Research Activities and Development Committees.”

National Information Management and Support System: View All Active Projects – Form 1002, United States Department of Agriculture, https://www.nimss.org/projects/active_list.

Frey M.W., Xiang C., Hoffman M.P., Taylor A.G., and Gardner J. 2009. Biodegradable chemical delivery system. USA Patent.

Halmer P. 2006. Quality. In: The Encyclopedia of Seeds – Science, Technology and Uses. Black, M., Bewley, J.D., and Halmer, P. (Eds.). p. 567. CAB International.

Madsen M.D., Petersen S.L. and Taylor A.G. 2010. Seed coating compositions and methods for applying soil surfactants to water repellent soil. USA Patent.

Perry S.E. and Yuan L. (eds) 2011. Plant Transcription Factors: Methods and Protocols, vol 754. Humana Press.

Pluskota W.E., Bradford K.J. and Nonogaki H. 2011. Tissue printing methods for localization of RNA and proteins that control seed dormancy and germination. In: Kermode A.R. (ed) Seed Dormancy: Methods and Protocols, Methods in Molecular Biology, vol 773. Springer, NY, pp 329-339.

Taylor A.G., Bolotin A., Pollicove S. and Taylor R. 2011. Controlled release of seed and soil treatments triggered by pH change of growing media. USA Patent.

TeKrony, D.M. 2006. Seeds: The delivery system for crop science. Crop Science 46:2263-2269.


Land Grant Participating States/Institutions


Non Land Grant Participating States/Institutions

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