Contents Summary655I.Definitions of stress656II.The seed life cycle revisited in view of the eustress–distress concept657III.Common denominators of many stresses: reactive oxygen and nitrogen species660IV.Alarm662V.Resistance664VI.Exhaustion666VII.Conclusions667Acknowledgements669References669 Summary ‘Stresses’ that impact upon seeds can affect plant reproduction and productivity, and, hence, agriculture and biodiversity. In the absence of a clear definition of plant stress, we relate concepts from physics, medicine and psychology to stresses that are specific to seeds. Potential ‘eustresses’ that enhance function and ‘distresses’ that have harmful effects are considered in relation to the seed life cycle. Taking a triphasic biomedical stress concept published in 1936, the ‘General Adaptation Syndrome’, to the molecular level, the ‘alarm’ response is defined by post‐translational modifications and stress signalling through cross‐talk between reactive oxygen and nitrogen species, and seed hormones, that result in modifications to the transcriptome. Protection, repair, acclimation and adaptation are viewed as the ‘building blocks’ of the ‘resistance’ response, which, in seeds, are the basis for their longevity over centuries. When protection and repair mechanisms eventually fail, depending on dose and time of exposure to stress, cell death and, ultimately, seed death are the result, corresponding to ‘exhaustion’. This proposed seed stress concept may have wider applicability to plants in general.

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SummaryMyrothamnus¯abellifolia, a short woody shrub from southern Africa, can survive severe desiccation of its vegetative organs. We studied mechanisms protecting this plant from oxidative damage during desiccation for 2 weeks, 4 and 8 months, and also during subsequent rehydration. This plant retains high concentrations of chlorophyll during desiccation, and these chlorophyll molecules are probably a source for potentially harmful singlet oxygen production. Desiccation triggered substantial increases in zeaxanthin and redox shifts of the antioxidants glutathione and ascorbate towards their oxidised forms. Simultaneously, the concentrations of violaxanthin, b-carotene, ascorbate, a-tocopherol, and glutathione reductase activity progressively decreased. Antheraxanthin, g-tocopherol, lutein, neoxanthin and glucose-6-phosphate dehydrogenase displayed less pronounced changes in response to desiccation. Even after 4 months of desiccation, Myrothamnus¯abellifolia recovered rapidly upon rehydration. Re-watering induced formation of ascorbate and glutathione, simultaneous reduction of their oxidised forms, and rapid production of a-tocopherol and of various carotenoids. Only after 8 months of desiccation did the antioxidant system of M.¯abellifolia break down; 3 weeks after the onset of rehydration, these plants abscised their leaves, but even then they were still able to recover and develop new ones. Ascorbate, b-carotene and a-tocopherol were totally depleted after 8 months of desiccation and did not recover upon rehydration; glutathione was partly maintained, but only in the oxidised form. We present a model demonstrating which parts of antioxidant pathways break down as oxidative stress becomes detrimental and we discuss some potential implications of our results for the genetic modi®cation of crop plants to improve their drought tolerance.

Assuming a fixed rate of desiccation, the higher water-holding capacities of hummock sphagna will allow them to continue metabolism for longer than other species. While this could be viewed as a form of 'desiccation avoidance', hummock species also recover faster than other species during rehydration, suggesting that they have higher inherent tolerance. This may help them to persist in drought-exposed hummocks. In contrast, species growing in wet habitats lack such strong avoidance and tolerance mechanisms. However, their turgor maintenance mechanisms, for example more elastic cell walls, enable them to continue metabolizing longer as their water contents fall to the turgor-loss point.

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Production of reactive oxygen species (ROS) is a widely

Recalcitrant seeds are intolerant of desiccation and cannot be stored in conventional seed banks. Cryopreservation allows storage of the germplasm of some recalcitrant seeded species, but application to a wide range of plant diversity is still limited. The present work aimed at understanding the stresses that accompany the first steps in cryopreservation protocols, wounding and desiccation, both of which are likely to lead to the formation of reactive oxygen species (ROS). Extracellular ROS production was studied in isolated embryonic axes of sweet chestnut (Castanea sativa). Axis excision was accompanied by a burst of superoxide (O(2)(*-)), demonstrated by a colorimetric assay using epinephrine, electron spin resonance and staining with nitroblue tetrazolium. Superoxide was immediately produced on the cut surface after isolation of the axis from the seed, with an initial 'burst' in the first 5 min. Isolated axes subjected to variable levels of desiccation stress showed a decrease in viability and vigour and increased electrolyte leakage, indicative of impaired membrane integrity. The pattern of O(2)(*-) production showed a typical Gaussian pattern in response to increasing desiccation stress. The results indicate a complex interaction between excision and subsequent drying and are discussed with a view of manipulating ROS production for optimisation of cryopreservation protocols.

Reactive oxygen species (ROS) are implicated in seed death following dehydration in desiccation-intolerant 'recalcitrant' seeds. However, it is unknown if and how ROS are produced in the apoplast and if they play a role in stress signalling during desiccation. We studied intracellular damage and extracellular superoxide (O2 · -) production upon desiccation in Castanea sativa seeds, mechanisms of O2 · -production and the effect of exogenously supplied ROS. A transient increase in extracellular O2 · -production by the embryonic axes preceded significant desiccation-induced viability loss. Thereafter, progressively more oxidizing intracellular conditions, as indicated by a significant shift in glutathione half-cell reduction potential, accompanied cell and axis death, coinciding with the disruption of nuclear membranes. Most hydrogen peroxide (H2O2)-dependent O2 · -production was found in a cell wall fraction that contained extracellular peroxidases (ECPOX) with molecular masses of~50 kDa. Cinnamic acid was identified as a potential reductant required for ECPOX-mediated O2 · -production. H2O2, applied exogenously to mimic the transient ROS burst at the onset of desiccation, counteracted viability loss of sub-lethally desiccation-stressed seeds and of excised embryonic axes grown in tissue culture. Hence, extracellular ROS produced by embryonic axes appear to be important signalling components involved in wound response, regeneration and growth.