Characterizing non-lethal damage within dry seeds may allow us to detect early signs of ageing and accurately predict longevity. We compared RNA degradation and viability loss in seeds exposed to stressful conditions to quantify relationships between degradation rates and stress intensity or duration. We subjected recently harvested (‘fresh’) ‘Williams 82’ soya bean seeds to moisture, temperature and oxidative stresses, and measured time to 50% viability (P50) and rate of RNA degradation, the former using standard germination assays and the latter using RNA Integrity Number (RIN). RIN values from fresh seeds were also compared with those from accessions of the same cultivar harvested in the 1980s and 1990s and stored in the refrigerator (5°C), freezer (−18°C) or in vapour above liquid nitrogen (−176°C). Rates of viability loss (P50−1) and RNA degradation (RIN⋅d−1) were highly correlated in soya bean seeds that were exposed to a broad range of temperatures [holding relative humidity (RH) constant at about 30%]. However, the correlation weakened when fresh seeds were maintained at high RH (holding temperature constant at 35°C) or exposed to oxidizing agents. Both P50−1 and RIN⋅d−1 parameters exhibited breaks in Arrhenius behaviour near 50°C, suggesting that constrained molecular mobility regulates degradation kinetics of dry systems. We conclude that the kinetics of ageing reactions at RH near 30% can be simulated by temperatures up to 50°C and that RNA degradation can indicate ageing prior to and independent of seed death.

Rapid, non-destructive methods for measuring seed germination and vigour are valuable. Standard germination and seed vigour were determined using 81 soybean seed lots. From these data, seed lots were separated into high and low germinating seed lots as well as high, medium and low vigour seed lots. Near-infrared spectra (950–1650 nm) were collected for training and validation samples for each seed category and used to create partial least squares (PLS) prediction models. For both germination and vigour, qualitative models provided better discrimination of high and low performing seed lots compared with quantitative models. The qualitative germination prediction models correctly identified low and high germination seed lots with an accuracy between 85.7 and 89.7%. For seed vigour, qualitative predictions for the 3-category (low, medium and high vigour) models could not adequately separate high and medium vigour seeds. However, the 2-category (low, medium plus high vigour) prediction models could correctly identify low vigour seed lots between 80 and 100% and the medium plus high vigour seed lots between 96.3 and 96.6%. To our knowledge, the current study is the first to provide near-infrared spectroscopy (NIRS)-based predictive models using agronomically meaningful cut-offs for standard germination and vigour on a commercial scale using over 80 seed lots.

Seed mass is an important plant functional trait linked to germination. For instance, higher-mass seeds often display greater germination compared to lower-mass seeds when exposed to non-stressful conditions. Yet, knowledge pertaining to germination dynamics for different mass-based seed fractions following exposure to abiotic stress is lacking. Here, we assess the germination response of relatively fresh, mass-separated Rudbeckia mollis (Asteraceae) seeds to various simulated seasonal temperatures, supra-optimal temperatures and increasing ageing stress duration. Air density separation yielded three mass-based classes, called light (393 ± 35 μg), intermediate (423 ± 29 μg) and heavy (474 ± 38 μg). Water uptake kinetics indicated that imbibition (0–6 h) and germination lag (6–24 h) were independent of seed mass. Similarly, germination and viability loss of fresh seeds following exposure to seasonal and supra-optimal constant temperatures were independent of mass. However, seed mass influenced germination following increasing ageing stress, with light seeds germinating to a significantly greater extent than intermediate or heavy seeds. For example, final germination per cent in light-class seeds was about 1.7 times greater than intermediate or heavy seeds after 20 d of saturated salt accelerated ageing (SSAA). Seeds stored for 1 year in the laboratory displayed mass-dependent germination patterns similar to seeds following SSAA. Mass-independent germination responses may be a strategy to maintain an annual life history in otherwise difficult environments when R. mollis seeds are relatively fresh. However, differences in germination response between aged and unaged seeds suggest that mass-dependent viability loss may occur in R. mollis.

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 within-species 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.

Embryonic axes of tea (desiccation sensitive) and pea (desiccation tolerant) were dried at different rates or stored at different water contents to distinguish between damage associated with the immediate effects of water loss and the longer-term effects of a partially hydrated state. No loss of viability was observed if pea axes were dried sufficiently rapidly (from 1.8 to 0.1 g H2O g-1dry mass (g/g) within 5 d). However, viability was lost in tea axes dried below 0.5 g/g (approximately -15 MPa) even if axes were dried within 1 h. Death in tea axes dried to moisture contents less than 0.5 g/g probably resulted from the removal of water necessary for cellular structural integrity (i.e. desiccation damage sensu stricto). When axes of both species were dried at slower rates, viability losses were observed at water potentials between about -3 and -15 MPa. The timing for this type of damage was species dependent, occurring within 2 d for tea and after 5 d for pea, and may be explained by higher oxidative activity in tea compared to pea. Embryos of both species with water potentials below -3 MPa were lethally damaged if oxygen consumption exceeded 1000–5000 μmol O2 g-1dry mass. Recalcitrant seeds are different than orthodox seeds because the former do not survive drying below a critical water content, regardless of the drying rate. Rapid drying is required for accurate assessment of the critical water content. Slow drying leads to metabolic imbalance and artefactual assessment of the critical water content for desiccation damage. Both tea and pea seeds were susceptible to damage from metabolic imbalances, suggesting that the predominant stress from slow drying is ageing.

The changes in water status of germinating and non-viable soybean (Glycine max L. Merr.) seeds were characterized by nuclear magnetic resonance (NMR) spectroscopy. There were distinct changes in water status between viable and non-viable soybean seeds. In dry seeds, there were only two components, bound and bulk water, as revealed by component analysis of NMR (T2) data. On the contrary, a three-component water proton system (bound, bulk and free water) was observed in both germinating and non-viable soybeans during Phase I of hydration. The bulk water component of non-viable seeds disappeared completely during the lag phase (Phase II) of hydration, resulting in a two-component water proton system. In contrast, the three-component water proton system in Phase II was observed in the germinating seeds. Rapid hydration (Phase III), following Phase II, was observed in germinating soybean seeds only. Due to reorganization of water protons, there was a concomitant increase in bulk and free water, but a decrease in bound water. The physical state of water in these seeds (analysed by NMR spectroscopy) and the measurements of tissue leachate conductivity suggest that non-viable soybean seeds were more affected by the disorganized cell structure in the seed membrane system. The present study also provides evidence that physical reorganization of water is essential in germinating soybean seeds during hydration.

Germination is an energy-demanding process that requires the operation of mitochondria, which must survive desiccation in the quiescent seed and become rapidly functional after imbibition to meet the ATP demand. The relationship between germination and mitochondrial performance was addressed by analysing the properties of mitochondria isolated from control, primed and aged pea (Pisum sativum L.) seeds. Mitochondria were isolated and purified at early stages of germination (before radicle protrusion), and their oxidative properties, membrane integrity and ultrastructure were examined. Mitochondria isolated after 12 h of imbibition readily oxidized exogenous NADH and Krebs cycle substrates at high rates. However, their phosphorylation efficiency was restricted by poor membrane integrity. After 22 h from the beginning of imbibition, purified seed mitochondria had intact outer membranes and oxidized the substrates at slightly lower rates, but with higher respiratory control (improved capacity for phosphorylation). Purified seed mitochondria were always found to be deficient in endogenous NAD, although the organelles were capable of importing and retaining the cofactor. While the priming treatment appeared to slightly increase the performance of mitochondria, seed deterioration by accelerated ageing strongly affected the oxidative properties of mitochondria, which were badly impaired in ATP production. Outer and inner membrane integrity was identified as the primary target for desiccation and ageing stress. A link between mitochondrial function and seed quality was also corroborated by respiration measurements of seed fragments at the onset of imbibition.

Seeds of watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai var. Congo) were subjected to mild accelerated ageing treatments of 30 and 60 d at 30°C at a moisture content of 7.2%. In comparison to a cold-stored control, normal germination and viability were slightly (but not significantly) decreased by these treatments. Subsequent construction of survival curves during storage at 40°C, at seed moisture contents ranging from 10–18%, revealed that the prior 30°C ageing treatments prolonged survival under these conditions.