Brief Summary of Minutes of Annual Meeting

Accomplishments: Objective 1. Identify and characterize biophysical, biochemical, genetic, and environmental factors regulating or influencing seed development, germination, vigor and dormancy. A graduate student, Steven Haba, has begun a project involved with seed production and characterization in Begonia. This effort includes development of rapid and efficient quantitative seed germination tests for the very tiny seeds; assessment of basic biochemical properties (carbohydrate, oil, protein content); desiccation tolerance (seeds of some species appear to be orthodox and relatively long-lived); and development of accelerated aging protocols for evaluation of vigor and prediction of long-term storage. Selected accessions of Rudbeckia fulgida are being grown during 2011 to produce fresh seed and begin a more detailed comparative study of GA treatment, and to combinations of GA and cold among the accessions. We are developing approaches for production of at least 10,000 seeds per accession of species that tend to flower over a long time and have a concurrent long fruit ripening period, that upon maturity results in ballistic dispersal of the 1 or 2 seeds per fruit. Muskmelon (Cucumis melo L.) is a warm-season, frost-sensitive crop that germinates poorly at low temperatures. Cold-sensitive cultivar Noy Yizre el (NY) and cold-tolerant cultivar Persia 202 (P-202) were compared at 15 and 25 C to determine how seed coat anatomy and oxygen uptake affected low-temperature germinability. The seed coats of both cultivars restricted oxygen uptake but NY had a lower rate of oxygen uptake compared to 'P-202' with or without the seed coat. The poor germination of NY at lower temperatures was related primarily to lower embryo oxygen uptake in addition to the seed coat restriction of oxygen uptake. The qSD1-2 and qSD7-2 seed dormancy QTLs were narrowed to short genomic regions of several candidate genes and the two QTLs were also associated plant height. The fine mapping data suggest that qSD12 may consists of more than one underlying gene. Some of the putative QTLs for seed dormancy or resistance to pre-harvest sprouting in wheat detected in the previous years were confirmed with the new data and the map positions were improved. Three QTL alleles enhancing resistance to pre-harvest sprouting were introduced to the same genetic background. We have demonstrated using functional gene transfer and RNAi silencing that a major quantitative trait locus (QTL) termed Htg6.1 associated with the ability for lettuce (Lactuca sativa) seeds to germinate at high temperatures (up to 37 C) is due to an allele of a gene (LsNCED4), which encodes an enzyme in the abscisic acid (ABA) biosynthetic pathway (9-cis-epoxycarotinoid dioxygenase 4). Lines having mutations in LsNCED4 were characterized and exhibit thermo-tolerance during germination. These lines are being backcrossed to purify from additional mutations and create isogenic lines. An additional strong Htg QTL has been mapped on chromosome 9 (Htg9.1) in a second RIL population derived from L. sativa accession PI251246 and cultivar Salinas. Fine mapping is in progress to identify candidate genes associated with this QTL. A new project has been initiated to identify genes associated with the phenotypic plasticity of lettuce seed dormancy and germination characteristics in relation to the seed maturation environment. Seeds of the parental lines of the RIL populations described above were produced in low (25 ºC) and high (35 C) temperature greenhouses. The seeds produced in the two environments exhibited distinctly different phenotypic germination responses to temperature and light. We will attempt to map the loci associated with this plasticity in the RIL populations derived from these parents and establish whether it is associated with epigenetic DNA markers (methylation). Foundation seed from the two RIL populations are being produced to enable their production in multiple environments to facilitate this mapping approach. Down regulating expression of thioredoxin in wheat improved certain starch paste traits of both wheat cultivars. Certain farinograph and extensograph properties of the high gluten variety did not change, while farinograph properties of the low gluten variety decreased slightly compared to the parental control. Thus achieving PHS tolerance through modulation of thioredoxin expression resulted in higher yields but did not come at the expense of dough quality. We devised two approaches to extract and identify proteins undigested by pepsin in uncooked and cooked meal from numerous sorghum and corn varieties. The first approach involves extracting residues undigested after 2 h with Borate-SDS-ME and separated by PAGE. Improved separation of undigested sorghum proteins was achieved using NuPAGE Bis-Tris gels. Western blots, with antibodies against particular zeins and kafirins, were used to monitor fates of different kafirins and revealed differential digestion rates. A second approach involved extracting undigested residues sequentially with 60% t-butanol, 60% t-butanol-ME and Borate-SDS-ME. The second approach, also coupled with western analysis, revealed the following. (i) Existence of oligomeric forms of certain kafirins that differ in the degree of their susceptibility to pepsin digestion. (ii) Effect of cooking on the formation and digestion of the oligomers. (iii) Cross-linked forms of most kafirins became more resistant to digestion after cooking. (iv) Most alpha-kafirins are preferentially extracted in 60% t-butanol-ME while most gamma-kafirins are extracted in Borate-SDS-ME buffer. (v) Monomeric kafirins are resistant to pepsin digestion. (vi) Gamma-kafirins form a series of oligomers that exhibit differential resistance to digestion. Our results suggest that this presently described systematic approach to analyzing the digestion by pepsin of sorghum prolamins should lead to greater insights into the digestion of specific types of sorghum grain proteins. Comparative proteomic analysis of recalcitrant and orthodox Spartina seeds, using two-dimensional electrophoresis coupled to mass spectrometry, is being conducted to identify proteins associated with desiccation tolerance, as well as during cold stratification to determine protein changes that occur during loss of dormancy. The breeding system of sugar beet does not facilitate traditional genetic approaches to gene discovery, so populations were constructed using self-fertility as a means to genetically dissect seedling vigor traits. A Rhizoctonia seedling disease nursery was continued, and results demonstrated resistance to Rhizoctonia damping-off is present in sugar beet, and resistance has been incorporated into the newly released enhanced germplasms SR98 (PI 655951) and SR98/2 (PI 659754). A candidate gene transcript profiling was untaken to examine early growth stages during germination. Genes involved in stress responses, signaling and growth were activated within one hour after imbibition. To see whether TERF1 up-regulates PR- and cell wall-associated genes in seeds, Arabidopsis transgenic plants, in which TERF is chemically inducible, have been generated using the GeneSwitch technology (see below). Twenty-four independent, transgenic lines, which are resistant to hygromycin, have been isolated. T-2 seeds have already been produced. The progeny plants are currently subjected to genotyping and gene expression analysis to examine responses to the chemical inducer, methoxyfenozide. Arabidopsis siliques on the maternal plants were treated with the chemical inducer to cause NCED6 induction in developing seeds. Comparison of NCED6 expression between ligand-treated and -untreated siliques in the same flowering stem indicated that NCED6 was specifically induced by the ligand application. Wild-type siliques and untreated transgenic siliques also showed NCED6 mRNA signals, however they do not indicate leaky expression but reflect the native expression of NCED6 during the maturation stages. This established system allowed us to plan on and propose more experiments to analyze molecular changes downstream, which has led to a successful funding from American Seed Research Foundation. The role of polycomb complex in the regulation of rice endosperm development and grain filling was examined in rice. We purified the rice OsFIE2-polycomb complex using tandem affinity purification and demonstrated its specific H3 methyltranferase activity. We found that the OsFIE2 gene product was responsible for H3K27me3 production specifically in vivo and the gene expression was not regulated by imprinting. Genetic studies showed that a severe reduction of OsFIE2 expression led to completely endosperm-free seeds and a moderated reduction of OsFIE2 expression resulted in smaller seeds and loss of seed dormancy. Genome wide ChIP-seq analyses found that a large number of endosperm specific regulatory genes and storage nutrient metabolic pathway genes were directly regulated by H3K27me3 modification in the rice endosperm. In a previous study, we found that high temperature germination sensitivity was genetically controlled by a major QTL on the short arm of chromosome 3A (Qhtgs.osu-3A) in two winter wheat populations of recombinant inbred lines (RILs) generated from the locally adapted cultivars, 'Intrada' x 'Cimarron' and 'Jagger' x '2174'. In the present study, we tested the same set RILs of the Jagger x 2174 population for response in germination to low temperature, high temperature, and normal temperature. Results showed that Qhtgs.osu-3A disappeared when the seeds of the population were treated at 4 C for 3-7 days or were kept at normal temperature at 25 C. We have also developed markers for three genes, ABC-3A, HSP16, and MFT-3A, and mapped them on the distal end of the short arm of chromosome 3A. The MFT1 was associated with Qhtgs.osu-3A. The hormone ABA triggers wheat grain dormancy. The problem was that wheat varieties sometimes lack seed dormancy leading to a tendency for mature grain to germinate on the plant before harvest. This preharvest sprouting causes financial hardship for farmers because the grain must be used for feed. A technique was developed to identify ABA hypersensitive mutations in wheat resulting in increased grain dormancy which should provide increased resistance to preharvest sprouting. The increase in ABA-hormone response caused grain to loose dormancy more slowly after maturity, but did not eliminate the capacity to loose dormancy by after-ripening. GA signaling controls seed germination and plant height. The problem is to understand the mechanisms by which the GA stimulates seed germination and stem elongation. Research showed that GA stimulates seed germination and plant growth by causing two F-box proteins, SLEEPY1 and SNEEZY, to target master negative regulators called DELLA for protein destruction. SNEEZY destroys the subset of DELLA proteins that control plant height, but not the DELLAs that control seed germination. ABA insensitive mutants of hard red spring Scarlet wheat showed decreased seed dormancy and rapid after-ripening. Scarlet ABA-insensitive lines show initial seed dormancy levels and preharvest sprouting tolerance comparable to those of the original wild-type Scarlet. However, ABA-insensitive lines after-ripened more rapidly requiring only 3 weeks rather than 4 months of after-ripening to germinate. New work is underway to define the occurrence of the respiratory transition during maize seedling growth and development. We have begun to characterize germination and seedling growth protocols in the laboratory under light and dark conditions with the goal of comparing temperatures to ascertain what happens to the respiratory transition during germination and seedling growth at low temperatures. Currently we are evaluating seed lots. The respiratory transition has been studied at temperatures around 22-25 C but we do not know of studies investigating the impact of low temperature stress on these parameters. Two research projects have been initiated to answer a pressing question for the seed industry: "Can soybean seeds maintain viability and vigor in storage after 1 year?" The farmers' demand for treated soybean seed has grown from less than 8% treated soybeans in 1996, to more than 30% in 2008 (Munkvold, 2009), and all indications are that the trend is continuing. Soybean seeds exposed to inadequate storage environments in dealers' warehouses and farmers' storage sheds can lose seed viability and vigor. In the past, seed companies sold the returned untreated seed from dealer's warehouses in the commodity market. Today, seed companies must incur high cost in disposing of treated soybean seed in an environmentally safe way. Prolonging soybean seed viability and vigor in storage could save seed companies the cost of seed disposal. 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. Relationships were developed with seed producers in the emerging wildflower seed industry of the southeast US and restoration/conservation practitioners to identify key areas of seed biology research. This is evidenced by $180,336 in funded grants spanning 2009-12. Projects revolved around developing models to describe desiccation sensitivity in palms, germination physiology of orchids, genetic diversity among seed producing populations of a key species used in wild land restoration, cryopreservation of dune-stabilizing plant germplasm, and assessing the wildflower seed industry in Florida. Obligate root parasitic plants of the Orobanchaceae do not germinate unless they chemically detect a host plant nearby. Some members of this family, like Orobanche, are noxious weeds that cause heavy crop damage to agriculture worldwide. The endothelium, which is the inner layer of the testa, rapidly absorbs water. Its interconnected cells contain labyrinthine walls and filled with mucilage, facilitating water accumulation for germination that starts after receiving germination stimuli. Swelling of the endothelium also leads to the opening of a water gap at the micropyle. The perisperm cells underneath this gap mediate between the rhizosphere and the embryo, and are the likely location for the receptors of germination stimuli. The other perisperm cells, as also the endosperm cells and parts of the embryo, are loaded with lipids and protein bodies. Many plasmodesmata connect the perisperm cells to each other, and the cells near the micropyle tightly surround the emerging seedling. These perisperm cells, and also the proximal embryo cells, resemble each other in their cytoplasmic contents, and are actively involved in transfer of reserve nutrients to the developing seedling during germination. We are collaborating in a workgroup on Germination, Trait Coevolution, and Niche Limits in Changing Environments sponsored by the National Evolutionary Synthesis Center (NESCent) at Duke University. Though this group, we are working with colleagues at the University of Arizona (Larry Venable) and at the Institute of Botany, Chinese Academy of Sciences (Zhenying Huang) to conduct hydrothermal time analyses of seed germination of 13 native or introduced species. We applied hydrothermal time analysis to seed germination responses in relation to after-ripening to assess how changes in dormancy relate to their ecological fitness and demographic success. A manuscript is in preparation describing this work, as is a review of applications of hydrothermal time analysis to seed ecology. Hydrothermal time analysis was also applied to characterize germination and dormancy properties of herbicide-susceptible and herbicide-resistant populations of Echinochloa phyllopogon in California. Applications of abscisic acid (ABA) have been investigated as a method to enhance drought tolerance and condition transplants to better withstand post-transplanting field stresses. We are now developing foliar application methods with ABA to improve transplant quality while suppressing stem elongation rates in vegetable transplants grown in the nursery. We examined ABA foliar applications on transplant growth and yield of jalapeno pepper and triploid watermelon. In pepper, ABA was applied once (3.8 mM) at 22, 15, or 8 days before transplanting (DBT) or twice (1.9 mM) at 22 and 15 DBT. In watermelon, ABA was applied in the similar timings but with half the rate used for pepper. ABA was also evaluated as a means to control growth of mature triploid watermelon transplants (5-, 6-, 7- and 8-week old). In pepper, plant height showed a similar reduction of 10 to 14% for all ABA treatments. In contrast, reductions in shoot dry weight varied from 17 to 35%, with the strongest inhibition induced by the single application at 22 DBT. In watermelon, ABA did not affect plant height, but reduced shoot dry weight by 16 to 23%. In both species, these differences diminished gradually after transplanting and no yield difference was observed. Objective 3. Develop, evaluate, and transfer technologies to assess and improve seed and seedling quality, health, performance, utilization, and preservation. Carbon nano-tubes (CNTs) and nanocrystals were characterized and then applied to seeds as a dry power (CNTs only) or dispersed in water/agar (both CNTs and nanocrystals). Research was initially conducted by applying CNTs to 3 crop seeds: tomato, cucumber and soybean. This crop selection provides one of each of three different permeability types, permeable, selective-permeable and non-permeable, in our lab's model system. We observed a germination enhancement in all seed types even at the lowest concentration of CNTs. We saw improved conditions on plates and in roll towels. This may be due to disinfecting/filtering properties of CNTs or their ability to eradicate pathogens and/or enhance germination. Next, we tested 4 crop seeds (by adding switchgrass) with nanocrystals. We sought to determine if this nano-size material that is smaller than CNTs could penetrate seed coats. We conducted a survey of all four crop seeds to determine if the 10 nm nanocrystals would penetrate the seed coat during imbibition. We are making more intensive use of our X-ray analysis capabilities to develop protocols and information about the quality of our seed accessions beyond germination tests (standard protocol for germplasm centers). We've been collaborating with Dr. Mark Roh of the USDA-ARS in evaluating seeds of Cornus and Corylopsis by X-ray imaging. A visiting scholar (V. Neumann-Silva) from USP-ESALQ (Brazil) did a careful analysis of seed free space and vigor in Portulaca. A manuscript based on this research is in preparation. Bacterial Fruit Blotch (BFB) in cucurbits is a disease caused by Acidovorax avenae affecting worldwide watermelon and melon production. Currently there is a zero tolerance policy for BFB in growing facilities; however, there are no assays sensitive enough to reliably detect BFB in seeds. Two of the possible options existing for detection of BFB in early stage development of infected plants are Fourier Transform Infrared Spectroscopy (FTIR) and the use of chemosensory analysis. FTIR analysis was conducted in dry seeds, imbibed seeds, germinating plants, and small seedlings in order to determine the limit of detection for the sensors. Measured spectrums and olfactory profiles will then be analyzed for differences. Early trials have demonstrated the feasibility of using FTIR to rapidly and nondestructively differentiate contaminated seeds from non-infected seeds. Also FTIR has nondestructively differentiated alive and dead seeds from within a seed lot. Recent human disease outbreaks of bacterial pathogens transmitted on vegetables, especially fresh spinach, have raised questions about the potential for seed transmission of human pathogens. The bacterial populations of spinach seed and seedling leaves were compared using DGGE, to assess bacterial community richness, and real-time PCR to compare the abundance of select phyla (total bacteria, Actinobacteria, Bacteroidetes, Firmicutes, a-Proteobacteria and b- Proteobacteria). To determine the effect of environment, the plants were grown in field and growth chambers. Vertical transmission of bacterial community members was evident because the developmental stage of the plant affected the richness and abundance of select bacterial phyla. The bacterial richness of plants grown in the two environments was not affected. However, a greater number of bacteria were associated with field grown samples in comparison to those produced in growth chambers. A statistically significant interaction existed between growth stage and environment for each of the selected phyla. Populations on cotyledons were smaller than mature leaves, but were not significantly different than the 3-4 leaf stage seedlings. The culturable populations of bacteria on seeds (~5 log CFU/g) were significantly smaller than determined using real time PCR (~7 log copies). Vertical transmission of bacteria from seeds to seedlings, suggests that seed may have evolved to ensure the transmission of certain bacteria. Switchgrass (Panicum virgatum L.) is used horticulturally as an ornamental and agronomically as an animal feedstock and a putative bio-energy crop. Genetic transformation, using somatic embryogenic (SE) callus derived from mature seeds, is one strategy for improving switchgrass traits. A superior switchgrass line, HR8, was developed in using recurrent tissue culture selection from cv. 'Alamo' that has low dormancy and produces a high percentage of somatic embryos. Eighty two percent of HR8 seeds germinated after harvest comparing to 26.8% for unselected 'Alamo'. HR8 seeds that germinated produced 84.9% SE callus. The respiratory activity of individual seeds during imbibition and germination is being assessed using the Q2 instrument from ASTEC Global. Initial seed respiration during imbibition and germination is being characterized in relation the effects of respiratory inhibitors to identify the metabolic pathways underlying different components of the oxygen uptake patterns exhibited by imbibed and germinating seeds. New methods were developed to analyze Q2 data that makes it amenable to use in population-based threshold models. We are using this approach to analyze and model respiratory responses to inhibitors, water potential, temperature and aging. The effects of seed drying methods on seed longevity are being assessed. Seeds exhibit hysteresis in their moisture content/relative humidity relationships, or isotherms, depending upon whether they are losing or absorbing water. Seeds that are drying have moisture contents that are generally 0.5 to 1% higher in moisture content than seeds that are hydrating to the same equilibrium RH from a lower moisture content. A second year of studies has confirmed that this difference in seed moisture content results in corresponding differences in seed longevity. Based on these results, simple modifications to seed drying procedures may be able to extend seed longevity in storage. With funding from the USAID-funded Horticulture Cooperative Research Support Program (HortCRSP), we are demonstrating and disseminating desiccant drying bead technology for seed drying and storage by smallholder farmers in India, Nepal, Bangladesh, Kenya, and Tanzania. Seed drying beads can dry seeds efficiently in hermetic containers, and can be regenerated by heat and reused indefinitely. This technology has considerable promise for improving seed quality and longevity in humid regions.

Argyris, J., Truco, M.J., Ochoa, O., McHale, L., Dahal, P., Van Deynze, A., Michelmore, R.W., Bradford, K.J. 2011. A gene encoding an abscisic acid biosynthetic enzyme (LsNCED4) collocates with the high temperature germination locus Htg6.1 in lettuce (Lactuca sp.). Theor. Appl. Genet. 122: 95-108. Bumgarner, N.R., M.A. Bennett, P.P. Ling, R.W. Mullen and M.D. Kleinhenz. 2011. Canopy cover and root-zone heating effects on fall and spring-grown leaf lettuce yield in Ohio. HortTechnology 21:737-744. Carson L., J. Freeman, K. Zhou, G. Welbaum, M. Reiter. 2011. Cultivar evaluation and lipid protein contents of Virginia grown edamame. HortTechnology. 21:131-135. Cerveny, C. B., W. B. Miller and A. G. Taylor. 2011. Storage temperature and moisture content affect respiration and survival of Ranunculus asiaticus dry tuberous roots. HortScience. 46: 1523-1527. Chappell J.H. and M.A. Cohn. 2011. Corrections for interferences and extraction conditions make a difference: use of the TBARS assay for lipid peroxidation of orthodox Spartina pectinata and recalcitrant Spartina alterniflora seeds during desiccation. Seed Sci. Res. 21: 153-158. Chappell J.H., and M.A. Cohn. 2011. Recalcitrant and orthodox Spartina seeds or isolated embryos exhibit similar leachate patterns immediately following desiccation (accepted pending revision: Seed Sci Res). Fernández-Arbaizar A, Nonogaki H, Sanz L, Lorenzo O. 2011. The clade-A protein phosphatase 2C HAB2 is an essential component of the ABA signalling pathway in Arabidopsis seeds. Informativo Abrates (Brazilian Association of Seed Technology) Special Issue, p.75. Gonzalez, M., L. Pollak, and S. Goggi. 2011. Genotype x environment interactions in populations possessing Ga1s and ga1 alleles for cross-incompatibility in maize. Euphytica. DOI 10.1007/s10681-011-0543-6. Gu, X.-Y., M. E. Foley, D. P. Horvath, J. V. Anderson, J. Feng, L. Zhang, C. R. Mowry, H. Ye, J. C. Suttle, K. Kadowaki, and Z. Chen. 2011. Association between seed dormancy and pericarp color is controlled by a pleiotropic gene that regulates ABA and flavonoid synthesis in weedy red rice. Genetics. 189:1515-1524. Hanson, L.E., Duckert, T.M., Goodwill, T.R., and McGrath, J.M. 2011. Beta PIs from the USDA-ARS NPGS evaluated for resistance to Cercospora beticola, 2010. Plant Disease Management Reports. 5:FC056. Hanson, L.E, McGrath, J.M. 2011. The perfect stage of powdery mildew (Erysiphe polygoni) of Beta vulgaris found in Michigan. Plant Disease 95:494. Hanson, L.E., and McGrath J. M. 2011. Rhizoctonia seedling disease on sugar beet. International Sugar Journal 113:584-589. Joel D., H. Bar, A. Mayer, D. Plakhine, H. Ziadne, J. Westwood, and G. Welbaum. Seed ultrastructure and water absorption pathway of the root-parasitic plant Phelipanche aegyptiaca (Orobanchaceae). Annals of Botany 2011; doi: 10.1093/aob/mcr261. Johnson, T.R., Kane, M.E., and Pérez, H.E. 2011. Examining the interaction of light, nutrients, and carbohydrates on seed germination and early seedling development of Bletia purpurea (Orchidaceae). Plant Growth Regulation. 63: 89-99. Kauth, P. and Pérez, H.E. 2011. Industry survey of the native wildflower market in Florida. HortTechnology. 21:779-788. Kikkert, J.R., G.S Abawi and A.G. Taylor. 2011. Seedling Establishment. Legume ipmPIPE Diagnostic Pocket Series. http://legume.ipmpipe.org. Kumar MB, A, Martin RC, and Nonogaki H. 2011. Isolation of microRNAs that regulate seed dormancy. In A.R. Kermode ed, Methods in Molecular Biology: Seed Dormancy, Humana Press, Totowa, 773: 199-213. Kuykendall, L.D., Shao, J., Naegele, R.P., and McGrath, J. M. 2011. Genetic disease resistance and conservation of a plant gene that encodes a mitogen-activated protein kinase kinase kinase. Advanced Studies in Biology 3:169-180. Leskovar, D.I., Agehara, S., Jifon, J., Crosby, K., Rush, C. and S. Goreta-Ban. 2011. Foliar ABA sprays controlled growth and improved survival and desiccation tolerance of vegetable transplants. Acta Horticulturae, 898: 237-244. Lopez-Velasco G., R. Boyer, G. Welbaum, and M. Ponder. 2011. Changes in spinach phylloepiphytic bacteria communities following minimal processing and refrigerated storage described using pyrosequencing of 16S rRNA amplicons. Journal of Applied Microbiology. 110 (5): 1203-1214. Lopez-Velasco G., G. Welbaum, and M. Ponder. 2001. Phyllopshere Bacterial Community Structure of Spinach (Spinacia oleracea) as Affected by Cultivar and Environmental Conditions at Time of Harvest. Diversity Journal. 3(4), 721-738; doi:10.3390/D3040721. Malone BM, Tan F, Bridges SM, Peng Z. 2011. Comparison of using rice endosperm H3K27 trimethylation profiling data. PLoS One. 6(9):e25260. Martinez-Andujar C, Martin RC, Bassel GW, Kumar MBA, Pluskota WE, and Nonogaki H. 2011. Post-transcriptional gene regulation during seed germination and stand establishment. 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Zhao. 2011. Selection and characterization of a new switchgrass (Panicum virgatum L.) line with high somatic embryogenic capacity for genetic transformation. Scientia Horticulturae 129(4): 854-861.