Dehydrins are a group of plant proteins that have been shown to be involved in the tolerance of various abiotic stresses such as dehydration, salinity, and low temperature. We have previously shown that the K-segments of the wheat dehydrin DHN-5 are essential for the protection of enzyme activities in vitro. In this study, we further investigate the role of the K-segments in the growth of Escherichia coli under various stresses, and we tested their antibacterial and antifungal activities. Our results showed that the truncated forms of DHN-5 containing the two K-segments enhanced tolerance of E. coli against diverse stresses by protecting proteins against aggregation. In addition, we demonstrated that the K-segments have antibacterial and antifungal activities against Gram-positive and Gram-negative bacteria and fungi. Based on these results, we propose that the K-segments may play a protective role in plants not only under abiotic stress conditions but also most likely during defense mechanisms.

Late Embryogenesis Abundant (LEA) proteins, a group of hydrophilic proteins, have been linked to survival in plants and animals in periods of stress, putatively through safeguarding enzymatic function and prevention of aggregation in times of dehydration/heat. Yet despite decades of effort, the molecular-level mechanisms defining this protective function remain unknown. In this paper, we summarize and review research discoveries of the classification of the LEA protein groups based on their amino acid sequence similarity and on the presence of distinctive conserved motifs. Moreover, we focus on high correlation between their accumulation and water deficit, reinforcing their functional relevance under abiotic stresses. We also discuss the biochemical properties of LEA proteins arising from their hydrophilic nature and by amino acid composition. Although significant similarities have not been found between the members of the different groups, a unifying and outstanding feature of most of them is their high hydrophilicity and high content of glycine. Therefore, we have highlighted the biotechnological applications of LEA genes, and the effects of over-expressing LEA genes from all LEA groups from different species of origin into different plant hosts. Apart from agronomical purposes, LEA proteins could be useful for other biotechnological applications in relation to their capacity to prevent aggregation of proteins.

Late Embryogenesis Abundant (LEA) proteins participate in plant stress responses and contribute to the acquisition of desiccation tolerance. In this report Rab28 LEA gene has been over-expressed in maize plants under a constitutive maize promoter. The expression of Rab28 transcripts led to the accumulation and stability of Rab28 protein in the transgenic plants. Native Rab28 protein is localized to nucleoli in wild type maize embryo cells; here we find by whole-mount immunocytochemistry that in root cells of Rab28 transgenic and wild-type plants the protein is also associated to nucleolar structures. Transgenic plants were tested for stress tolerance and resulted in sustained growth under polyethyleneglycol (PEG)-mediated dehydration compared to wild-type controls. Under osmotic stress transgenic seedlings showed increased leaf and root areas, higher relative water content (RWC), reduced chlorophyll loss and lower Malondialdehyde (MDA) production in relation to wild-type plants. Moreover, transgenic seeds exhibited higher germination rates than wild-type seeds under water deficit. Overall, our results highlight the presence of transgenic Rab28 protein in nucleolar structures and point to the potential of group 5 LEA Rab28 gene as candidate to enhance stress tolerance in maize plants.

LEA (late embryogenesis abundant) proteins participate in plant stress tolerance responses, but the mechanisms by which protection occurs are not fully understood. In the present work the unfolded proteins from maize dry embryos were analyzed by mass spectrometry. Twenty embryo proteins were identified, and among them 13 corresponded to LEA-type proteins. We selected three major LEA proteins, Emb564, Rab17 and Mlg3, belonging to groups 1, 2 and 3, respectively, and we undertook a comparative study in order to highlight differences among them. The post-translational modifications of native proteins were analyzed and the anti-aggregation properties of recombinant Emb564, Rab17 and Mgl3 proteins were evaluated in vitro. In addition, the protective effects of the LEA proteins were assessed in living cells under stress in Escherichia coli cells and in Nicotiana bentamiana leaves agroinfiltrated with fluorescent LEA-green fluorescent protein (GFP) fusions. Protein visualization by confocal microscopy indicated that cells expressing Mg3-GFP showed reduced cell shrinkage effects during dehydration and that Rab17-GFP co-localized to leaf oil bodies after heat shock. Overall, the results highlight differences and suggest functional diversity among maize LEA groups.

Group-2 late embryogenesis abundant (LEA) proteins, also known as dehydrins, are claimed to stabilize macromolecules against damage caused by freezing, dehydration, ionic or osmotic stresses. However, their precise function remains unknown. Here, we investigated the effect of wheat dehydrin (DHN-5) protein on the activity and thermostability of two distinct enzymes, beta-glucosidase (bglG) and glucose oxidase/peroxidase (GOD/POD) in vitro. The purified DHN-5 protein had the capacity to preserve and stabilize the activity of bglG subjected to heat treatment. In addition, DHN-5 stabilized oxidizing enzymes, as it improved reliability in measuring glucose concentrations with a glucose oxidase/peroxidase (GOD/POD) kit while the temperature increased from 37 to 70 degrees C. All together the data presented provide evidence that DHN-5 is a dehydrin able to preserve enzyme activities in vitro from adverse effects induced by heating.

Salinity is one of the severest environmental stresses affecting plant productivity. In many plant species, salt sensitivity is associated with the accumulation of sodium (Na + ) in photosynthetic tissues. Here, we provide the physiological and molecular analyses of seedlings of two Tunisian durum wheat genotypes ( Triticum turgidum L. subsp. Durum [Desf.]), Mahmoudi (salt sensitive) and Om Rabia3 (salt tolerant). Na + and K + contents in leaf sheath from Om Rabia3 were significantly higher than those of Mahmoudi. However, the net uptake of Na + from the soil occurred at similar rates in both varieties, suggesting that Om Rabia3 has much stronger ability to limit Na + flux from roots to leaf blades. This mechanism could be explained by a capacity of Om Rabia3 to retain higher Na + concentration in leaf sheath and unload less Na + from the xylem to the upper shoots. When treated with 100 mM NaCl leaf sheaths of Om Rabia3 developed lower water potentials and a higher relative water contents than those of Mahmoudi. These features may arise from enhanced osmotic adjustment in Om Rabia3. Measurements of stomatal conductance, free proline and chlorophyll content also indicate that Om Rabia3 is better adapted to tolerate high salt than Mahmoudi. A correlation was obtained between the expression pattern of TaSOS1 (a plasma membrane Na + /H + antiporter) in the roots and sheaths of both wheat varieties and the Na + fluxes from roots to leaves. TaSOS1 transcript accumulated in Mahmoudi than in Om Rabia3, suggesting repression of TaSOS1 in the tolerant variety that reduces loading of Na + to the upper shoots. These results help to design new genetic screens for salt tolerance in wheat.

Late Embryogenesis Abundant (LEA) proteins are associated with tolerance to water-related stress. A wheat (Triticum durum) group 2 LEA proteins, known also as dehydrin (DHN-5), has been previously shown to be induced by salt and abscisic acid (ABA). In this report, we analyze the effect of ectopic expression of Dhn-5 cDNA in Arabidopsis thaliana plants and their response to salt and osmotic stress. When compared to wild type plants, the Dhn-5 transgenic plants exhibited stronger growth under high concentrations of NaCl or under water deprivation, and showed a faster recovery from mannitol treatment. Leaf area and seed germination rate decreased much more in wild type than in transgenic plants subjected to salt stress. Moreover, the water potential was more negative in transgenic than in wild type plants. In addition, the transgenic plants have higher proline contents and lower water loss rate under water stress. Also, Na(+) and K(+) accumulate to higher contents in the leaves of the transgenic plants. Our data strongly support the hypothesis that Dhn-5, by its protective role, contributes to an improved tolerance to salt and drought stress through osmotic adjustment.