Background and Aims Determining seed longevity by identifying chemical changes that precede, and may be linked to, seed mortality, is an important but difficult task. The standard assessment, germination proportion, reveals seed longevity by showing that germination proportion declines, but cannot be used to predict when germination will be significantly compromised. Assessment of molecular integrity, such as RNA integrity, may be more informative about changes in seed health that precede viability loss, and has been shown to be useful in soybean. Methods A collection of seeds stored at 5 °C and 35–50 % relative humidity for 1–30 years was used to test how germination proportion and RNA integrity are affected by storage time. Similarly, a collection of seeds stored at temperatures from −12 to +32 °C for 59 years was used to manipulate ageing rate. RNA integrity was calculated using total RNA extracted from one to five seeds per sample, analysed on an Agilent Bioanalyzer. Results Decreased RNA integrity was usually observed before viability loss. Correlation of RNA integrity with storage time or storage temperature was negative and significant for most species tested. Exceptions were watermelon, for which germination proportion and storage time were poorly correlated, and tomato, which showed electropherogram anomalies that affected RNA integrity number calculation. Temperature dependencies of ageing reactions were not significantly different across species or mode of detection. The overall correlation between germination proportion and RNA integrity, across all experiments, was positive and significant. Conclusions Changes in RNA integrity when ageing is asymptomatic can be used to predict onset of viability decline. RNA integrity appears to be a metric of seed ageing that is broadly applicable across species. Time and molecular mobility of the substrate affect both the progress of seed ageing and loss of RNA integrity.

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HighlightSpecies, storage products, and moisture have large effects on the nature and quantity of volatile emission from dry seeds, but storage time and seed viability do not.

The duration that seeds stay vigorous during storage is difficult to predict but critical to seed industry and conservation communities. Production of volatile compounds from lettuce seeds during storage was investigated as a non-invasive and early detection method of seed ageing rates. Over 30 volatile compounds were detected from lettuce seeds during storage at 35 degrees C at water contents ranging from 0.03 to 0.09 g H(2)O g(-1) dw. Both qualitative and quantitative differences in volatile composition were noted as a function of water content, and these differences were apparent before signs of deterioration were visible. Seeds stored at high water content (L >or=0.06 g H(2)O g(-1) dw) emitted molecular species indicative of glycolysis (methanol+ethanol), and evidence of peroxidation was apparent subsequent to viability loss. Seeds containing less water (0.03-0.05 g H(2)O g(-1) dw) produced volatiles indicative of peroxidation and survived longer compared with seeds stored under more humid conditions. Production of glycolysis-related by-products correlated strongly with deterioration rate when measured as a function of water content. This correlation may provide a valuable non-invasive means to predict the duration of the early, asymptomatic stage of seed deterioration.

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Quercus species adapted to arid and semi-humid climates still produce recalcitrant seeds. The ability to avoid freezing rather than drought may be a more important selection factor to increase desiccation tolerance. Cryopreservation of recalcitrant germplasm from temperate species is currently feasible, whilst additional protective treatments are needed for ex situ conservation of Quercus from tropical and subtropical areas.

Seed shelf-life or longevity is difficult to predict or to measure on a practical time scale. Predictive models suggest that water has the same effect on ageing rate for all seed lots within a species and that initial seed quality is the dominating factor explaining withinspecies variation. These assumptions are used in 'accelerated ageing' or 'controlled deterioration' (AA/CD) tests, which are commonly used to predict seed longevity in commercial and research applications. In this study, we describe within-species variation of longevity for seeds of Secale cereale and S. strictum (cultivated rye and its wild progenitor) under typical dry storage conditions of a genebank, and show that initial seed quality is an important, but not sole, factor explaining measured longevity. We also test the correlation of seed longevity, measured under humid and dry conditions, using 50 cultivars of rye, wheat (Triticum aestivum) and the intergeneric cross triticale, to assess how well AA/CD tests predict seed shelf-life during dry storage. Known differences in longevity between wheat and rye were confirmed at all water contents, and triticale seeds demonstrated intermediate behaviour. Longevity measured for humid and dry conditions were weakly correlated when comparisons included all grain types and were not correlated in within-grain-type comparisons. Response to moisture varied among cultivars. These findings do not support assumptions made in seed ageing models that use AA/CD tests. Our results suggest that more traits are involved in the expression of seed longevity than those typically measured in studies of initial seed vigour.