Accumulation of Dehydrins and ABA-Inducible Proteins in Wheat Seedlings during Low-Temperature Acclimation

Using one-dimensional SDS-PAGE and immunochemical methods, we detected the presence and estimated the content of dehydrins and ABA-inducible (RAB) polypeptides in etiolated seedlings of four spring and three winter wheat (Triticum aestivum L.) cultivars differing in frost hardiness. We hardened three-day-old seedlings at 4°C for nine days or grew them at 22°C for a day (control seedlings). We established that heat-stable cold-regulated (COR) polypeptides with mol wts of 209, 196, 169, 66, 50, and 41 kD, which are characteristic of hardened wheat seedlings, were homologous to polypeptides from a dehydrin family and polypeptides with mol wts of 209, 196, 66, 50, and 41 kD were immunologically related to RAB-proteins. We supposed that these COR polypeptides were involved in the prevention of local protein dehydration and denaturation during hypothermia. Analysis of the relative content of COR proteins revealed a close correlation between the cultivar frost hardiness and the concentration of these proteins. It seems evident that different accumulation of dehydrins and RAB polypeptides in different cultivars of a single species is one of the causes for different plant frost hardiness.     

Association of dehydrins with wheat mitochondria during low-temperature adaptation

In mitochondria from the crowns of field-grown winter wheat plants or their seedlings hardened in the laboratory, thermostable proteins immunologically related to dehydrins were detected. It was found that two dehydrins with mol wts of 63 and 52 kD bound with the outer mitochondrial membrane during autumnal hardening or during adaptation to low temperature in the laboratory. Dehydrins of similar mol wts were detected among proteins in the total membrane fraction from low-temperature-adapted wheat plants. In addition, dehydrins with mol wts of 209 and 196 kD were present in this fraction as well. Dehydrins of similar mol wts were bound with mitochondria from seedlings adapted to low temperature and those from the crowns of plants after autumnal hardening. In spring, the amount of dehydrins associated with mitochondria from the crowns declined to the level characteristic of early autumn. Dehydrin association with mitochondria is evidently an important defense mechanism of frost-resistant plants.Translated from Fiziologiya Rastenii, Vol. 52, No. 2, 2005, pp. 221–226.Original Russian Text Copyright © 2005 by Borovskii, Stupnikova, Antipina, Anuchina, Voinikov.This revised version was published online in April 2005 with a corrected cover date.     

蛋白质的变化与植物抗寒性的关系研究进展

蛋白质的变化在植物抗寒生理研究中一直被广泛关注。低温胁迫期间在蛋白质含量变化的同时,还可能发生质的变化,合成新的蛋白质——低温诱导蛋白。综述了低温胁迫期间植物体内蛋白质的变化,重点阐述了抗冻蛋白、脱水蛋白和热激蛋白等3种低温诱导蛋白的特性及其与植物抗寒性的关系,并对该领域今后的研究做了展望,为进一步阐明植物抗寒的分子机制、提高植物的抗寒力提供了新的思路。     

Seasonal patterns of growth, dehydrins and water-soluble carbohydrates in genotypes of Dactylis glomerata varying in summer dormancy

• Background and Aims Summer dormancy in perennial grasseshas been studied inadequately, despite its potential to enhanceplant survival and persistence in Mediterranean areas. The aimof the present work was to characterize summer dormancy anddehydration tolerance in two cultivars of Dactylis glomerata(dormant ‘Kasbah’, non-dormant ‘Oasis’)and their hybrid using physiological indicators associated withthese traits. • Methods Dehydration tolerance was assessed in a glasshouseexperiment, while seasonal metabolic changes which produce putativeprotectants for drought, such as carbohydrates and dehydrinsthat might be associated with summer dormancy, were analysedin the field. • Key Results The genotypes differed in their ability tosurvive increasing soil water deficit: lethal soil water potential(_s) was –3·4 MPa for ‘Kasbah’ (althoughnon-dormant), –1·3 MPa for ‘Oasis’,and –1·6 MPa for their hybrid. In contrast, lethalwater content of apices was similar for all genotypes (approx.0·45 g H₂O g d. wt^–1), and hence the greater survivalof ‘Kasbah’ can be ascribed to better drought avoidancerather than dehydration tolerance. In autumn-sown plants, ‘Kasbah’had greatest dormancy, the hybrid was intermediate and ‘Oasis’had none. The more dormant the genotype, the lower the metabolicactivity during summer, and the earlier the activity declinedin spring. Decreased monosaccharide content was an early indicatorof dormancy induction. Accumulation of dehydrins did not correlatewith stress tolerance, but dehydrin content was a function ofthe water status of the tissues, irrespective of the soil moisture.A protein of approx. 55 kDa occurred in leaf bases of the mostdormant cultivar even in winter. • Conclusions Drought avoidance and summer dormancy arecorrelated but can be independently expressed. These traitsare heritable, allowing selection in breeding programmes.     

Differential accumulation of water stress-related proteins, sucrose synthase and soluble sugars in Populus species that differ in their water stress response

Proteins inducible by dehydration and abscisic acid (ABA), have been identified in a number of species and have been suggested to play a role in desiccation tolerance. Recently, we identified a novel boiling-stable protein (BspA) which accumulated in shoots of aspen ( Populus tremula L.) cultured in vitro, in response to gradual water stress and ABA application (Pelah et al. 1995. Tree Physiol. 15: 673–678.). Accumulation of BspA, and of the water stress-related protein dehydrin dsp- 16 and sucrose synthase from the resurrection plant. Craterostigma plantagineum , was examined in two greenhouse-grown Populus species to investigate the relationship between the presence of the proteins and water stress tolerance. Detached leaves of Populus tomentosa lost more water than Populus popularis , resulting in a significant decrease in leaf water potential. Using electrolyte leakage analysis, it was found that detached leaves of Populus popularis are more tolerant to water stress than those of Populus tomentosa . Using western blots with the corresponding antibodies, we have found in Populus popularis accumulation of BspA and sucrose synthase due to water stress, and the constitutive presence of a dehydrin-like protein. In contrast, a low expression of BspA was found in Populus tomentosa , but not of sucrose synthase and dehydrin-like proteins. Desiccation tolerance in many tissues can be partly attributed to soluble sugars. Analysis of the amount of soluble sugars did not reveal clear-cut differences between the two species, except for significant sucrose accumulation and glucose reduction in water-stressed Populus tomentosa and increase in glucose in water-stressed Populus popularis . The data obtained points to a positive correlation between increased water stress tolerance of one poplar species as compared with another and accumulation of water stress-related proteins and sucrose synthase.     

Water-stress-induced heat tolerance in geranium leaf tissues: A possible linkage through stress proteins?

Evidence is accumulating in favor of a linkage at the cellular level between various abiotic stresses. We conducted a study to evaluate the effect of water stress on the heat tolerance of zonal geraniums, Pelargonium × hortorum cv. Evening Glow. Water stress was imposed by withholding irrigation until pots reached 30% (by weight) of well‐watered controls, and by maintaining the pot weight by additions of water for another 7 days. Leaf xylem water potential (XWP, MPa), relative water content (RWC. %), and heat‐stress tolerance (HST; LT₅₀, defined as the temperature causing half‐maximal % injury based on electrolyte leakage) were measured in control, stressed, and recovered plants. Proteins were extracted from the leaves following the above treatments, and SDS‐PAGE and immunoblotting were performed by using standard procedures. Immunoblots were probed with antibodies to dehydrin and 70‐kDa heat shock cognate (HSC70) proteins. Data indicate that XWP and RWC, respectively, were −0.378 MPa and 92.3% for control plants and −0.804 MPa and 78.6% for stressed plants. Water‐stressed plants exhibited a significant increase in HST compared to control (LT₅₀ of 55°C vs 51°C). Water‐stress‐induced HST was not due to heat acclimation (leaf warming in stressed plants). Data also indicate that water‐stress treatment did not increase freezing tolerance of geranium leaves. Increased HST was associated with the accumulation of several heat‐stable, dehydrin proteins (25–60 kDa), and both cytosolic and ER luminal (BiP) HSC70 proteins. Leaf XWP, RWC, and HST reversed to control levels concomitant with the disappearance/reduction of dehydrins and HSC70 proteins in water‐stress‐relieved plants. The possibility of a cellular linkage between water stress and heat‐stress tolerance is discussed.     

Seasonal Changes in the Composition and Content of Dehydrins in Winter Wheat Plants

Seasonal changes in the pattern and content of dehydrins in winter wheat (Triticum aestivum) plants grown under field and laboratory conditions were studied by one-dimensional PAGE and immunochemical methods. During hardening, plants accumulated dehydrin-like polypeptides with mol wts of 209, 196, 66, 50, and 41 kD. In winter, low-molecular-weight dehydrins with mol wts of 24, 22, 17, 15, and 12 kD were synthesized and accumulated as well. Their content dropped sharply in spring when plants became unhardened. Accumulation/disappearance of these proteins corresponded to the fluctuations in wintering plant frost tolerance before winter and in spring. It is assumed that both high- and medium-molecular-weight dehydrins are involved in plant stress responses and adaptation, whereas low-molecular-weight dehydrins are evidently involved only in the process of low-temperature adaptation.     

Abscisic Acid and Desiccation Tolerance in Mosses

In higher plants the phytohormone ABA is involved in processes that are connected to water deficit, like stomatal closure or desiccation tolerance. In bryophytes, also containing ABA in their tissues, physiological functions remained uncertain for a long time. Quite recently, several papers have shown different effects of exogenously applied ABA: stomatal closure in Anthoceros, drought hardening in Funaria and production of the landform in Riccia. In all these cases the relevant conditions (water deficit) enhance the endogenous ABA level significantly. For induced desiccation tolerance, ABA serves as a mediator to induce specific proteins (dehydrins) strongly connected with this tolerance. Therefore, it can be concluded that in bryophytes ABA has the same function as in higher plants. It acts as a mediator in stress conditions.