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Jian Fang and E.E. Roos

Achieving the optimum moisture content for long-term seed storage usually requires that seeds be dried after receipt at a genebank. Soybean (Glycine max L.) seeds were dried using four procedures: over concentrated H2S O4, over silica gel, at 15% relative humidity (RH), or in an oven at temperatures of 30, 35 and 40C. Following dying seeds were stored at 40C for 10 days and at 5C for one yr. Seeds were evaluated for germination and vigor (root length, dehydrogenase, and leachate conductivity). Initial moisture content (mc) was reduced from 8.3% to between 6.6% (24 hr at 30C) and 4.6% (H2S O4, 30 days). Germination and vigor of seeds was essentially unchanged immediately following the drying treatments. Storage for 10 days at 40C reduced germination by up to 12% while storage for one yr at 5C had a similar effect (14% maximum loss) for most treatments. The treatments having the lowest drop in germination after one yr of storage treatment were the silica gel and the 30C oven treatments, which dropped only 3% in germination. Drying at 15% RH, also resulted in a lower loss in germination. In all three tests, vigor of seeds after storage at 40C was higher than controls for the the silica gel and 15% RH treatments as well as for the 30C and 35C oven treatments. Storage at 5C gave similar results for all three vigor assessments.

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Sharon Sowa and Eric E. Roos

Infrared spectroscopy was used to measure biochemical changes during bean (Phaseolus vulgaris L.) seed imbibition. Transmission spectroscopy of excised embryonic axes revealed changes in lipid phase (gel to liquid crystalline) and protein secondary structure within the first 15 min of hydration. Spectral changes in seed coats, cotyledons, and axes during the first 2 hr of imbibition (measured in vivo) were detected using photoacoustic sensing. Onset of seed respiration could be detected as early as 15 min after addition of water. CO2 production, demonstrated by the appearance of a double peak centered at 2350 cm-1, increased with time of imbibition. Infrared photoacoustic spectroscopy of intact seeds holds promise as a method for non-invasive viability assessment.

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Ming Zhang and Eric E. Roos

All kinds of plant seeds evolve volatile compounds during storage. However, a reliable deterioration forecast method is still not established using volatile evolution, even though some preliminary work indicated a relationship between volatile evolution and seed deterioration (Fielding and Goldsworthy, 1982; Hailstones and Smith, 1989; Zhang et al., 1993). Here we review some of the previous work concerning seed volatiles and present some more recent research on the effects of seed moisture content on deterioration. We found that volatile evolution from seeds was controlled by seed moisture level. Generally, seeds tended to evolve more hexanal and pentanal under extremely dry conditions (below 25% equilibrium RH). The production of hexanal and pentanal decreased with increasing seed moisture level. On the other hand, methanol and ethanol increased with increasing seed moisture. All of the volatile compounds accumulated in the headspace of the seed storage container during storage. Therefore, it should be possible to use different volatiles to indicate the deterioration of seeds stored under different moisture levels. We suggest that hexanal may be used for seed assessing deterioration under dry storage conditions (below 25% equilibrium RH), while ethanol may be used for seeds stored under higher moisture conditions (above 25% equilibrium RH). [References: Fielding, J.L. and Goldsworthy, A. (1982) Seed Sci. Technol. 10: 277–282. Hailstones, M.D. and Smith, M.T. (1989) Seed Sci. Technol. 17: 649–658. Zhang et al. (1993) Seed Sci. Technol. 21:359–373.]

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F. D. Moore III and E. E. Roos

An index “internal slope” derived from the cumulative frequency distribution of individual seed leachate conductivities is related to seed quality; the larger the index value the less variation among individual seeds in a sample (100 seeds) and the higher the seed quality. We have recently developed data acquisition/instrurment control/data smoothing/data analysis software which accesses frequency and cumulative frequency distributions of individual seed conductivities and the derived index on an almost continuous basis from the start of the first soaking.

At present, lack of convergence with regard to curve fitting may occur necessitating multiple sampling times. A “window in time” approach is described whereby index estimates during a two-hour interval within the index stability phase are averaged. Evidence of the method's ability to assess seed vigor will be presented.

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E.E. Roos and D.A. Davidson

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Eric E. Roos and Loren E. Wiesner

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Sharon Sowa, Eric E. Roos, and Francis Zee

Seeds of the recalcitrant species lychee (Litchi chinénsis Sonn.) and longan (Dimocarpus longan Lour.) were stored near 100% relative humidity at 8 to 10C in air, 80% nitrous oxide (N2O) plus 20% oxygen, or 100% nitrous oxide. The combination of anesthetic and oxygen extended storage longevity of both species. Seeds stored in 100% N2O lost terminability at the same rate as those stored in air. Lychee seeds retained 92% of initial germination after 12 weeks under 80% N2O/20% O2, while those under air retained only 44%. Longan seeds failed to germinate after 7 weeks under air, yet retained 70% of their initial germination under 809” N2O/20% O2. The combination of anesthetic and oxygen atmospheres could provide a new approach to recalcitrant seed storage.

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Eric E. Roos, Sharon Sowa, and Kristin Pedas

Short term soaking of seeds does not appear to be detrimental to seed viability and may provide a means of testing seed viability non-destructively. Seeds of corn (Zea mays L.) and rice (Oryza sativa L.), differing in viability, were soaked for 0.5, 1, 2 and 4 hr in distilled water at room temperature. Analyses of pH, protein/polypeptides (BCA assay and absorbance at 280 nm), and potassium (and other metals), were done on individual seed leachates. After each time period seeds were germinated for 7 d to determine viability. For both corn and rice, pH remained constant between 0.5 and 4 hr of soaking. Protein concentration gradually increased during the 4 hr soak in both corn and rice, but varied with seed lot. Potassium was the most common metal excreted and increased 3 to 4 fold between 0.5 and 4 hr of soaking. Although seed to seed variability in any given lot was high, in general, low viability seeds lost more cellular constituents than high viability seeds.

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Sharon Sowa, Eric E. Roos, and Francis Zee

Seeds of the recalcitrant species Litch i chinénis and Euphoria longan were stored in humid conditions at 8-10C under three different atmospheres: air, 80% nitrous oxide (N20)/20 % oxygen, and 100% nitrous oxide. The combination of anesthetic and oxygen extended storage longevity of both species. Oxygen was required for maintenance of viability; seeds stored under 100% N20 lost germinability at the most rapid rate. Lychee seeds retained 92% of control germination after 12 weeks under 80% N20/20% 02, while those under air lost 56% viability. Longan seeds lost all viability after 7 weeks under air, yet retained 70% of their control germination under 80% N20/20% 02. The combination of anesthetic and oxygen atmospheres could provide a new approach to recalcitrant seed storage.

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Sheila A. Blackman and Eric E. Roos

The low quality of some seed lots received by germplasm repositories such as the National Seed Storage Laboratory can thwart efforts to regenerate seed for storage. This germplasm is in danger of irretrievable loss. The aim of this work is to promote the germination, and hence regeneration, of such low quality seeds through sterile culture of the isolated embryos. Hybrid (B73×LH51) maize seeds were aged 5 y at 32°C and 0.037 g H2O g-1 dry wt. Vigor - but not viability -declined under these conditions. The effects of four factors on growth and germination were systematically examined. These were: seed pretreatments; antibiotics and fungicides; nutrients; and growth substances. Amongst the pretreatments, none surpassed partial hydration of seeds for 24 hr to 0.55 g H2O g-1 dry wt at 25°C prior to embryo dissection. Thiram (2.4 mg mL-1) and kanamycin (50 ug ml1) effectively controlled bacterial and fungal growth with no deleterious effects on growth during culture of the isolated embryos. Exogenous sucrose (optimum 5 % wt/vol) significantly stimulated radicle growth in both deteriorated and non-deteriorated embryos. No other organic or inorganic nutrient stimulated growth. Naphthalene acetic acid did not affect growth while kinetin reduced radicle growth and stimulated coleoptile growth. Gibberellic acid (GA3 at 10-5M) significantly stimulated radicle growth in deteriorated embryos, whereas it promoted coleoptile growth in both deteriorated and non-deteriorated embryos. These data suggest GA or a GA-stimulated process may limit the growth of aged embryos.