Brief Summary of Minutes of Annual Meeting

April 25 8:30 a.m. Susana Goggi: calls meeting to order, presents opening remarks, requests participants introduction, and requests President Xingyou Gu to chair the meeting. 8:40 a.m. Xingyou Gu: 1) Approval of 2012 Minutes (Kent Bradford motion, Hector Perez 2nd, and all in favor). 2) Hector Perez (the past president) provided a brief report on the state of W-2168 membership. New possible members were discussed. 3) Leadership structure: Kent leads the Secretary-elect Nominating Committee; current president, Xingyou Gu; president-elect, Susana Goggi; Secretary, Pablo Jourdan. 4) Discussion of possible seed biology symposium: after review of W-3168 proposal and further discussion on sites and other seed symposia currently on the schedule for the next few years, no actions were taken. 5) International Innovation Report from Research Media Ltd. was discussed and a decision was made not to take action. 6) Manjit Misra welcomed the group. 7) Donn Thill, Administrative Advisor, joined the meeting via phone link: he complemented the group regarding the proposal re-write and discussed next steps in the review and renewal process. 8) Hans Arne Jensen shared his views regarding the status of ISTA laboratories and Seed Testing in Europe; 11:00 a.m. State reports started: Dr. Alan Taylor and Dr. Kent Bradford presented their annual reports. Questions and discussions were made after each of the report. 2:00 p.m. State reports continued: Dr. Robert Geneve and Dr. Greg Welbaum presented their annual reports. Questions and discussions were made after each of the reports. 3:00 p.m.: Tour of Seed Science Center at Iowa State University. April 26 8:30 a.m.: Xingyou calls meeting to order. Welcome by Dr. Joe Colletti, Senior Associate Dean, ISU College of Agriculture and Life Sciences, and Associate Director of the Experiment Station. 9:00 a.m. State reports continued: Drs. Susana Goggi, Hector Pérez, Dr. Pablo Jourdan, Bruce Downie, Sabry Elias, and Xingyou Gu presented their annual reports. Questions and discussions were made after each of the reports. 11:30 am.: Reports of Secretary-elect, site location meeting for 2014 annual meeting, and other new business. Bob Geneve was named as the incoming Secretary. Meeting site for 2014 annual meeting will be at Ohio State University in July. 12:15 pm: Adjourn.

Objective 1. Identify and characterize biophysical, biochemical, genetic, and environmental factors regulating or influencing seed development, germination, vigor and dormancy Bradfords group (CA) cloned a QTL associated with high temperature germination (htg) in lettuce, which was identified as the gene LsNCED4 encoding the enzyme 9-cis-epoxycarotenoid dioxygenase 4 in the abscisic acid (ABA) biosynthetic pathway. An additional htg QTL was mapped on chromosome 9 (Htg9.1). Fine mapping of the new QTL is in progress to identify candidate genes. An atlas of gene expression during imbibition and germination of four genotypes of lettuce under different temperature and ABA conditions is being developed using RNA sequencing (RNASeq). The seed dormancy gene DELAY OF GERMINATION 1 (DOG1) in Arabidopsis has been determined to be involved in high temperature germination and the DOG1 homologs have been isolated from lettuce to characterize their expression and function(s) in lettuce seed dormancy. Pérezs group (FL) focused on seed development, dormancy, and germination ecology of wildflowers and some warm-season grasses. Seed development studies investigated desiccation tolerance, germination ability, and aging stresses in developing seeds and embryos. Seed dormancy studies addressed dormancy developmental controls, dormancy mechanisms expressed by seeds, and methods to alleviate these mechanisms. Germination ecology research revolved around germination timing and the seeds interaction with the environment to alleviate dormancy and promote germination. Nonogakis group (OR) performed research on mechanisms of tissue specific gene expression in tomato seeds. GeneChip analysis was used to identify multiple genes whose expression was enriched in the micropylar region of endosperm (endosperm cap) in tomato seeds. Some of these genes contained the conserved DNA motifs, suggesting their regulation by a common regulatory protein. The TOMATO ETHYLENE RESPONSE FACTOR1 (TERF1) was identified as a potential regulatory protein that enhances expression of multiple endosperm cap-specific genes. Function of hormone metabolism genes in Arabidopsis has been characterized. Gus group (SD) continued the research on seed dormancy and also initiated a project on seed longevity in rice. The project of cloning the qSD12 QTL was focused on developing isogenic lines for the natural alleles of three candidate genes and transgenic T2-T4 lines with the candidates. Haplotypic analysis for the qSD12 region was conducted for 400 lines of wild, weedy and cultivated rice. Genetic and physiological analyses were conducted for two chromosomal segments each containing the QTL clusters qSD1-2/qPH1 and qSD7-2/qPH7 for seed dormancy/plant height. QTL and haplotype analyses for seed dormancy and its related adaptive traits were conducted for U.S. weedy rice populations. Seed longevity was assessed for a segregating population of seeds aged under the room condition for about 10 years. A new population of >400 recombinant inbred lines derived from a line of U.S. weedy rice was advanced to the F8 generation to investigate ecological and genetic mechanisms of seed longevity under controlled and field conditions. Baskins group (KY) investigated mechanisms of breaking seed dormancy in various species including Solanum rostratum, Cardiocrinum giganteum var. giganteum, Alyssum minus, Astazgalus arpilobus and Suaeda corniculata, gave eight lectures on seed dormancy and seed bank, and published 18 papers with various collaborators. Downies group (KY) characterized the role of CRYPTOCHROME 1 and 2 in Cryptochome control of blue light induced dormancy in barley and assessed the function of the ATP-dependent, DEAD-box, RNA-Helicase PRH75 in seed viability and morphology. Geneves group (KY) worked on Eastern gamagrass, a warm-season, perennial grass native to large areas across North America. Plantings are commonly established from seed, but seed quality is a barrier to establishment. Standard laboratory purity methods and germination testing protocol needs to be standardized for routine assessment of eastern gamagrass seed lots for planting purposes. A survey of 21 labs determined that at least 9 germination procedures are currently in use. A comparison of a single seed lot as reported by 10 labs was used to determine variation in test results. Additional seed lots were germinated on substrates with and without potassium nitrate (KNO3) and subjected to prechilling and presoaking treatments. The portion of the seed quality and assessment portion of the gamagrass work has been completed. Work is now beginning of the treatments for improved dormancy release and increased first year stand. Stebers group (WA) worked on the wheat preharvest sprouting problem and mechanisms of the plant hormone GA in seedling growth and development. ABA-insensitive mutants in the genetic background of the dormant red wheat cultivar Scarlet were characterized. Cohns group (LA) continued working on physiological/molecular mechanisms of seed dormancy, seed desiccation intolerance (recalcitrance) and related problems in the marsh grass. Spartina alterniflora is an important component for successful maintenance and remediation of Louisiana coastal stabilization. Differentially expressed proteins between the dormant and nondormant state of Spartina alterniflora and Spartina pectinata seeds were isolated by two-dimensional gel electrophoresis comparisons and sequenced using mass spectrometry. The effects of drying rate and drying temperature on recalcitrant seed death will be re-examined. Jangs lab (OH) has been looking at the involvement of tandem CCCH zinc finger proteins (TZFs) in hormone-mediated developmental and environmental responses. Several TZFs were characterized for expression profile during seed germination in Arabidopsis and their responses to the ABA and GA hormones. Mercers lab (OH) has been exploring the germination characteristics of different landraces of maize from southern Mexico to determine how each might respond to potentially altered climatic conditions. Jourdans lab (OH) at the Ornamental Plant Germplasm Center (OPGC) continued several projects. One project was to study the biology of Begonia seeds and to develop an appropriate germplasm collection that emphasizes seed storage. Efforts have made to develop rapid and efficient quantitative seed germination tests for the minute seeds; assess basic biochemical properties (carbohydrate, oil, protein content) and desiccation tolerance (seeds of some species appear to be orthodox and relatively long-lived), and develop accelerated aging protocols for evaluation of vigor and prediction of long-term storage. The other project was to develop efficient seed production protocols for the 200 accessions of Phlox that have been accumulated and maintained as plant samples over the past two years. Controlled pollination techniques and seed dormancy characteristics in P. paniculata were investigated. The OPGC has also been exploring the germination patterns of different species of Rudbeckia. Obendorfs lab (NY) conducted research on several projects to determine if changes in soluble carbohydrates in hypocotyl and radicle tissues correlate with differential tissue survival after drying seedlings in buckwheat (Fagopyrum esculentum Moench) embryos, if raffinose and stachyose are unloaded from seed coat to developing embryos in soybean, and whether increasing the supply of free cyclitols to immature embryos increases accumulation of galactosyl cyclitols in soybean lines selected for low raffinose and low stachyose (LRS) or low raffinose, low stachyose and low phytin (LRSP1, LRSP2) concentrations in mature seeds, as compared with a CHECK line with normal raffinose, stachyose and phytin. Objective 2. Determine and model the biotic and abiotic factors affecting seed germination, seedling emergence, and establishment of sustainable populations in natural and agro-ecological systems Bradfords group (CA) collected accessions of Brachypodium hybrida, which is a moderately invasive, introduced grass species in California, and all were determined to be polyploid, although both diploid and tetraploid forms are present in its native range in Turkey. The collections from the state and the accessions from Turkey and other locations are being used to study the inherent, variation or plasticity in flowering time and seed dormancy to test the hypothesis that polyploids may have greater genetic variability in the flowering and seed dormancy traits, which may explain the preponderance of polyploids among invasive species. Pérezs group (FL) developed models to describe desiccation sensitivity in palms, cryopreservation of dune-stabilizing plant germplasm, examined tolerance of U. paniculata seeds to various concentrations of a commercial biocide, evaluated the chronic and range-wide loss of seed viability loss in Aristida stricta, and assessed cryo-storage tolerance of 4 U. panicualta seed accessions. Baskins group (KY) investigated dormancy cycling in the cold desert ecosystem and soil seed banks in forests of northern Iran, and helped pull together the literature on the ecological importance of mucilage in seed germination ecology. Downies group (KY) assessed the homologs of Arabidopsis and soybean LATE EMBRYOGENESIS ABUNDANT (LEA) protein client proteins, identified preferred LEA binding partners, characterized three SEED MATURATION PROTEIN1 (SMP1) binding proteins for functions in inducing thermodormancy in Arabidopsis, and constructed a web page as a data repository as part of a project examining Arabidopsis and soybean LEA protein interacting client proteins. McGraths group (MI) has focused on stress responses of seed germination and seedling field emergence/persistence in sugar beets to develop healthy, vigorous, and productive field populations of the crop. Despite planting high-quality, technically-augmented seed for growers with very high germination (>92%), field emergence and persistence continues to hover at ~60% in Michigan. Research has generated four transcriptome datasets of a high vigor variety germinated in stressful environments, including water, hydrogen peroxide, sodium chloride, and hydrogen peroxide plus sodium chloride. From over 200 million sequences assessed, and in comparison to water, each treatment showed between 75 and 100 statistically significant, differentially expressed genes, with approximately 10% overlap between any pair of treatments, and only seven shared among treatment comparisons. Approximately 50% of these have no descriptors yet found in nucleotide sequence databases, suggesting germination genes are less well represented in comparative genomics resources. Welbaums group (VA) worked on Bacteria Fruit Blotch (BFB), a seed-transmitted disease of cucurbits caused by Acidovorax avenae subsp citrulli (Aac) that is one of the most devastating diseases that affects both watermelon (Citrullus lanatus) and cantaloupe (Cucumis melo) production worldwide. The genome of Aac strain AAC00-1 (group II) was sequenced. The cucurbit germplasm from the USDA collect was screened to identify genetic resistance to different type III effectors. The group was also involved in a project to assess the potential for biological contamination on greenhouse lettuce and basil plants grown in aquaponic wastewater. Objective 3. Develop, evaluate, and transfer technologies to assess and improve seed and seedling quality, health, performance, utilization, and preservation. Bradford group (CA) has developed new methods to analyse Q2 data to monitor the respiratory activity of individual seeds during imbibition and germination, which makes it amenable to use in population-based threshold models. Effects of respiratory inhibitors on seed oxygen consumption during imbibition and germination have been characterized. Research has been started to identify metabolites and metabolic pathways underlying different components of the oxygen uptake patterns exhibited by imbibed and germinating seeds. The effects of seed drying methods on seed longevity are being assessed. The Q2 instrument has been utilized to show that seed respiration rates are highly correlated with germination rates (timing). Semi-automated respiratory assays in the Q2 can be used to collect germination rate data without requiring labor-intensive repeated observations of germination tests. With funding from the USAID-funded Horticulture Cooperative Research Support Program (HortCRSP), a desiccant drying bead technology was developed for seed drying and storage by smallholder farmers in India, Nepal, Bangladesh, Kenya, and Tanzania. The knowledge and techniques developed by this group were presented in five seminars, in addition to publications. Nonogakis group (OR) developed a gene switch system for the Arabidopsis NCED gene, which provided a proof of concept for modification of seed performance by hormone metabolism engineering. While the Gene Switch system is useful for agriculture, chemical application may not be practical for large-scale agriculture. Therefore, a novel system that could cause spontaneous seed dormancy that does not require chemical induction has also been tested. Considering potential technology development as an outcome of this study, the concept has been tested using genes isolated from wheat and sorghum. The NCED gene, one of the rate-limiting genes for ABA biosynthesis, has been isolated from sorghum and fused with the promoter sequence of EARLY METHIONINE-LABELED, one of seed-specific and ABA-regulated genes, from wheat. Arabidopsis plants expressing this chimeric gene have been generated and tested for seed germination performance. Gus group (SD) continued the project to pyramid the dormancy-enhancing alleles at four QTL detected/reported in the previous research to improve the resistance to pre-harvest sprouting for local spring wheat cultivars with white grain color. Taylors group (NY) focused on mechanisms and technology development of breaking seed dormancy in switchgrass (Panicum virgatum). A major limitation of growing switchgrass is the poor and erratic stands of the crop during the first year due to seed lot quality and dormancy. A series of experiments were conducted to examine effects of the plant hormones/growth regulators GA, ABA, ACC and fluridone on germination capability. Research on seed priming using the varieties Kanlow and Blackwell was conducted to develop a technology to promote germination, especially under environmental stress. Welbaums group (VA) worked on the development of several germination techniques. Basil is a popular herb with little information about its base, ceiling and optimal germination temperatures. Six different basil genotypes were compared for responses to high, low, and optimum temperatures. Thermogradient tables were used to generate a wide range of temperatures for seed testing on a flat surface. New thermogradient tables were designed. A preliminary gradient table has been compared for germination performance of a wide range of peanut genotypes. FTIR (Fourier transform infrared spectroscopy) is a technique, which is used to obtain an infrared spectrum of absorption, emission, photoconductivity or Raman scattering of a solid, liquid or gas. An FTIR spectrometer simultaneously collects spectral data in a wide spectral range. A mathematical algorithm converts the raw data into the actual spectrum. This technology has been evaluated for its possible use for the rapid non-destructive assessment of seed and plant disease infection. The other activities included teaching a web-based, asynchronous distance learning Vegetable Seed Production Course and organizing the XXVIII International Horticultural Congress on Science and Horticulture for People (IHC2010) in Lisbon, Portugal. Goggis group (IA) continued seed physiology and pathology programs. The seed physiology program can be organized into two areas of research: how to select, breed, and produce soybean and corn cultivars with enhanced nutritional properties and other value-added traits and high seed quality; and how to minimize adverse effects of production environments on seed quality and storage life of soybean and corn. Current projects include genome fluidity and stress-induced changes in the soybean genome and their effect on seed and agronomic characteristics; seed storage in soybean and the effect of soil fertility and the use of seed treatment to prolong seed life. The seed pathology research program can be organized into three areas: integrated management of multiple pathogens and pests attacking seeds and seedlings; seed health test development; and infection of corn by mycotoxin-producing fungi. Current disease management projects include: investigations into various aspects of soybean seedling blight and root rot caused by Fusarium species; management of multiple maize and soybean seedling pathogens through seed treatment combinations; and the evaluation of seed treatment effects on maize head smut through a PCR-based seedling assay. The development of seed health test method has been focused on seed-borne diseases of edible beans (bacterial wilt and Fusarium wilt) and tomatoes (Potato spindle tuber viroid and other viroids). Mycotoxin-related projects include assessing the role of mycotoxins in seedling diseases of maize, soybean, and wheat; and evaluation of the impact of transgenic insect resistance on the quality of maize grains for ethanol production. Other activities include leadership of the distance-education Graduate Program in Seed Technology and Business, and seed health testing standardization efforts through the US National Seed Health System. Jourdan and Bennetts group (OH) worked on developing protocols and information about the quality of seed accessions in the Ornamental Plant Germplasm Center beyond germination tests (standard protocol for germplasm centers) by more extensively use of X-ray analysis capabilities. Lopes and Bennetts group (OH) assessed seed germination and vigor from different cultivars of snap beans (Phaseolus vulgaris L.) varying in seed coat color and seed size after being submitted to drum priming and subsequent drying treatments. SVIS and other seed quality tests (including seedling emergence in greenhouse and field studies) have been conducted. Leskovars group (TX) investigated effects of plant hormones on seedling establishment and quality. Exogenous abscisic acid (ABA) was applied to induce leaf chlorosis in various vegetable and ornamental crops, a potential problem for its commercial use. One mechanism generally proposed for ABA-induced chlorosis is senescence by stimulated ethylene production. ABA accumulation during water stress also inhibits leaf expansion to limit plant water loss. When this acclimation is induced by exogenous ABA, it is followed by rapid leaf expansion, with leaf area eventually recovering to the control level, as shown in the previous research. The group further investigated the effect of ABA on Arabidopsis leaves at different stages of development and expansion to determine if the hormone inhibits cell expansion and/or cell division. Research was also conducted to determine the relation between chlorophyll loss and ABA concentrations in muskmelon seedlings.

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