Dehydrins are late embryogenesis abundant (LEA) proteins that accumulate during seed maturation and in response to abiotic stresses in vegetative tissues. They are thought to protect cellular components from dehydration stress. However, whether they play a role in survival in the dry state is not clear. In this study, an RNA interference (RNAi)-construct against the seed-expressed dehydrin of Arabidopsis thaliana, LEA14 (At2g21490), was introduced to wild-type plants, which led to a strong reduction in transcript abundance of the target gene as well as that of two other seed-expressed dehydrin homologues, XERO1 (At3g50980) and RAB18 (responsive to abscisic acid 18, At5g66400) in the transformants. Mature, dry seeds from the RNAi plants germinated to at least 95% after rehydration, indicating that seed desiccation tolerance was not affected, while they exhibited a twofold reduction in longevity. When stored at 75% relative humidity and 35°C, the seeds of two independent RNAi lines lost 50% of their viability in 10 d and 5 d, respectively, while it took 17 d for wild-type seeds to lose 50% viability. In addition, when seeds were imbibed in the presence of 100 mM NaCl, the seeds of RNAi plants exhibited reduced germination compared to wild-type seeds, suggesting that at least one of the three seed-specific dehydrins plays a role both against deterioration during storage at low moisture content and when imbibed tissues are submitted to salt stress at high moisture.

Dehydrin and QP47, proteins present in mature pea seeds (Pisum sativum), have been proposed to play protective roles during desiccation. To identify possible relationships between these proteins and desiccation tolerance, their tissue locations and patterns of synthesis and degradation have been examined during germination. Tissue locations were determined by immunocytochemistry using polyclonal antibodies raised against a conserved dehydrin amino acid sequence and against purified QP47. In embryonic axis and cotyledon cells, QP47 and dehydrin were distributed uniformly with no apparent nuclear or organellar specificity. Both proteins were present in 24 h-imbibed axes that had not initiated radicle growth but were completely absent from 24 h-imbibed axes that had begun to grow. The amounts of QP47 and dehydrin in embryonic axes decreased with time after the start of imbibition and were undetectable by 48 h. When germination was prevented by polyethylene glycol (PEG) or abscisic acid (ABA), both proteins remained at their original amounts. Thus, both QP47 and dehydrin disappeared coincidently with the beginning of growth and not simply as a function of the time after imbibition. QP47 persisted in cotyledons until at least 31 days into seedling growth, whereas dehydrin was not detectable in cotyledons after 7 days. Dehydrin, but not QP47, could be re-induced in pea shoots and cotyledons by dehydration. The timing of degradation of both proteins was correlated with the loss of desiccation tolerance during germination of pea axes.

Plants undergo a series of physiological, biochemical and molecular changes in response to adverse environmental conditions or stresses such as drought, low temperature or high salt. Several genes and their corresponding proteins have been described that may play a role in withstanding water-deficit-related stresses or full desiccation. In particular, sugars and late-embryogenesis-abundant (LEA) proteins have received the most attention. Plant responses to water-deficit and desiccation have been well-characterized at the molecular level; however, pinpointing the precise roles of the gene products in protecting cells under conditions of water deficit remains a challenging task. While few plants are capable of withstanding full desiccation, most seeds undergo this event as a pre-programmed and final stage in their development. These are the so-called ‘orthodox’ seeds. In contrast to seeds of orthodox species, those of recalcitrant species do not acquire desiccation tolerance during their development and are shed from the parent plant at relatively high water contents. The essential components of desiccation tolerance of seeds are likely to involve the ability to effect repair upon subsequent rehydration as well as the ability to accumulate protective substances that limit the amount of damage which otherwise would be caused by water loss. Studies have begun to examine whether the desiccation sensitivity of recalcitrant seeds is at least partially the result of an insufficient accumulation of LEA-type proteins, or whether other factors (including a lack of protective sugars) are more important. This review assesses some of these studies as well as recent research to understand gene and protein function using transgenic host plant systems.

The presence of dehydrins could not be demonstrated in axes of mature, undried recalcitrant seeds of the tropical wetland species Avicennia marina, Barringtonia racemosa, Bruguiera exaristata, Bruguiera cylindrica, Bruguiera gymnorrhiza, Ceriops tagal, Rhizophora apiculata, Rhizophora mucronata and Rhizophora stylosa, but were present in the temperate species Acer saccharinum, Aesculus hippocastanum, Araucaria angustifolia, Camellia sinensis, Castanea sativa, Poncirus trifoliata and Zizania palustris. They were also present in axes of Castanospermum australe (of tropical origin) seeds which underwent development in a temperate climate, and were produced in response to drying in axes of Barringtonia racemosa but not Avicennia marina. The presence of dehydrins was associated with high abscisic acid contents. These proteins may provide protection against low temperatures in temperate seeds and against water loss to which the seeds may be naturally exposed. The presence of dehydrins was unrelated to the evolutionary status of the families studied.

Western blot analysis using antiserum raised against the lysine-rich box common to dehydrins, one class of late-embryogenesis-abundant (LEA) proteins, was used to study the abundance of heat-stable ‘dehydrin-like’ proteins during development and water stress in Ranunculus sceleratus L. achenes (seeds). A 61 kDa dehydrin-like protein was apparently limited to immature seeds (fresh and dried) which had not attained full desiccation tolerance. In contrast, lower-molecular-mass proteins which were induced by desiccation were found only in more mature seeds. The molecular masses of desiccation-induced proteins changed during seed development from 18 kDa in seeds harvested at 13 days post anthesis (DPA) to 31 kDa at harvest maturity, 21 DPA.

Placing seeds at 21 DPA in polyethylene glycol (PEG) at −1.5 MPa reduced seed moisture content and was accompanied by accumulation of 31 kDa protein. This protein was no longer detected when the seeds were transferred to water. In seeds harvested at 13 DPA, PEG induced the synthesis not only of 18 kDa protein (which is associated with dried seeds at this developmental stage), but also of 28 kDa and 31 kDa proteins. These dehydrin-like proteins were also synthesised when seeds at 13 DPA were imbibed in water. These and other data indicate that both quantitative and qualitative changes in dehydrin-like proteins can occur in R. sceleratus, depending on seed maturity and the degree and duration of water stress.

Swards of cocksfoot (cvs KM2, Lutetia) and perennial ryegrass (cvs Aurora, Vigor) were grown under full irrigation or severe (80 d) drought in a field experiment in the South of France. Responses of the bases of immature leaves plus enclosed tissues were made during the drought period and after rewatering.

By the end of the drought, water content had fallen from 3·0 to 0·8 gwater g−1dm, and osmotic potential from −1·0to −4·5 MPa in all cvs. Measured minerals and water-soluble carbohydrates contributed, respectively, c 19 and 44% to osmotic potential in droughted leaf bases. The drought-sensitive cocksfoot cv. Lutetia was characterized by a large proportion of fructans having a low degree of polymerization (DP=3, 4). As drought progressed, accumulation of dehydrin transcripts and ABA were higher in leaf bases of the sensitive cv. Lutetia than in the resistant cv. KM2.

After rewatering, the water status of immature leaf bases returned to control levels in 1–2 d, and then increased further as leaves began to grow and new tissue was produced. High-DP-fructans remained unchanged in leaf bases of ‘Lutetia’ but were depleted by over 55%, and therefore remobilized, in leaf bases of other cvs after 8 d.

It is concluded that enclosed immature leaf bases survive drought by tolerating a low water status and that changes conventionally associated with desiccation tolerance are expressed most strongly in susceptible plants least able to maintain their water supply.

Dehydrin proteins (late embryogenesis abundant (LEA) D11 family) are produced in a wide variety of plant species in response to environmental stimuli with a dehydrative component, including drought, low temperature, salinity, and developmental stages such as seed and pollen maturation. Despite their widespread occurrence and abundance in cells under dehydrative conditions, the biochemical role of dehydrins remains elusive. The subcellular location of dehydrins is consistent with a biochemical role as an intracellular stabilizer, possibly with surfactant characteristics, acting upon targets in both the nucleus and cytoplasm. In some species, dehydrin loci are located within quantitative trait loci (QTL) intervals for important phenotypic traits including winter hardiness in barley (Hordeum vulgare L.) and anthesis-silking interval in maize (Zea mays L.). Dehydrin loci tend to be multigenic and occur in clusters on more than one chromosome. Investigations are currently under way in our laboratory and others' to move beyond protein accumulation studies and correlations with QTL to uncover direct cause-and-effect relationships between dehydrin (dhn) genes and phenotypes associated with physiological responses to stress.