The thioredoxin h-type TdTrxh2 protein of durum wheat confers abiotic stress tolerance of the transformant Arabidopsis plants through its protective role and the regulation of redox homoeostasis.

The thioredoxins (Trxs) are ubiquitous and they play a crucial role in various biological processes like growth and stress response. Although the functions of Trxs proteins are described in several previous reports, the function of the isoform Trxh2 of durum wheat (Triticum durum L.), designated as TdTrxh2, in abiotic stress response still unknown. Thus, we aimed in this study the functional characterization of TdTrxh2 through its expression in yeast cells and Arabidopsis plants. Sequence analysis revealed that TdTrxh2 protein shared the conserved redox site with the other Trxh from other plant species. Under various abiotic stresses, TdTrxh2 was up-regulated in leaves and roots of durum wheat. Interestingly, we demonstrated that TdTrxh2 exhibit protective effect on LDH activity against various treatments. Besides, the expression of TdTrxh2 in yeast cells conferred their tolerance to multiple stresses. Moreover, transgenic Arabidopsis expressing TdTrxh2 showed tolerance phenotype to several abiotic stresses. This tolerance was illustrated by high rate of proline accumulation, root proliferation, low accumulation of reactive oxygen species like H2O2 and O2·-, and high antioxidant CAT and POD enzymes activities. All these findings suggested that TdTrxh2 promotes abiotic stress tolerance through the redox homoeostasis regulation and its protective role.

Identification of a putative kinase interacting domain in the durum wheat catalase 1 (TdCAT1) protein.

Catalases are crucial antioxidant enzymes that regulate plants responses to different biotic and abiotic stresses. It has been previously shown that the activities of durum wheat catalase proteins (TdCAT1) were stimulated in the presence of divalent cations Mn2+, Mg2+, Fe2+, Zn2+, and Ca2+. In addition, TdCAT1s can interact with calmodulins in calcium-independent manner, and this interaction stimulates its catalytic activity in a calcium-dependent manner. Moreover, this activity is further enhanced by Mn2+ cations. The current study showed that wheat catalase presents different phosphorylation targets. Besides, we demonstrated that catalase is able to interact with Mitogen Activated Proteins kinases via a conserved domain. This interaction activates wheat catalase independently of its phosphorylation status but is more promoted by Mn2+, Fe2+ and Ca2+ divalent cations. Interestingly, we have demonstrated that durum wheat catalase activity is differentially regulated by Mitogen Activated Proteins kinases and Calmodulins in the presence of calcium. Moreover, the V0 of the reaction increase gradually following the increasing quantities of Mn2+ divalent cations. Such results have never been described before and suggest i) complex regulatory mechanisms exerted on wheat catalase, ii) divalent cations (Mn2+; Mg2+; Ca2+ and Fe2+) act as key cofactors in these regulatory mechanisms.

Superoxide dismutase (SOD) family in durum wheat: promising candidates for improving crop resilience.

The SOD family has been extensively analyzed at genome wide level in several crops. However, little is known about this family in durum wheat. In this study, a total of 14 TdSOD genes were identified in whole durum wheat genome including 8 TdCu-ZnSODs, 2 TdMnSODs, and 4 TdFeSODs. In silico analysis evinced that TdSOD family members displayed a closer evolutionary relationship, similar gene structure and protein features with their homologs from other plant species. Furthermore, the analysis of their promoter regions revealed the presence of a great number of cis-regulatory elements related to plant development, abiotic and biotic stresses, phytohormones, and several potential binding sites for transcription factors. Interestingly, 3D structure analysis revealed that TdCu-ZnSOD2A-2 and TdCu-ZnSOD2B-2, belonging to the Cu-Zn group, were modeled as copper chaperone for SOD like their homologs from rice and Arabidopsis. The expression profile of eight TdSOD candidate genes was investigated under salt, drought, cold, and ABA treatments. Notably, TdCu-ZnSOD2A-1, TdFeSOD4A-1, and TdFeSOD7A-1 were significantly up-regulated under all stress treatments. On the other hand, TdCu-ZnSOD7B and TdMnSOD2B were strongly expressed in roots and leaves under cold stress and TdCu-ZnSOD2B-2 was particularly up-regulated in leaves under ABA treatment. Ultimately, these findings provide valuable information for the identification of attractive candidate genes to improve wheat resilience.

The Putative Auto-Inhibitory Domain of Durum Wheat Catalase (TdCAT1) Positively Regulates Bacteria Cells in Response to Different Stress Conditions.

Catalase is a crucial enzyme in the antioxidant defense system protecting organisms from oxidative stress. Proteins of this kind play important roles in controlling plant response to biotic and abiotic stresses by catalyzing the decomposition of H2O2. The durum wheat catalase 1, TdCAT1, has been previously isolated and characterized. Here, using bio-informatic analysis, we showed that durum wheat catalase 1 TdCAT1 harbors different novel conserved domains. In addition, TdCAT1 contains various phosphorylation residues and S-Nitrosylation residues located at different positions along the protein sequence. TdCAT1 activity decreased after treatment with λ-phosphatase. On the other hand, we showed that durum wheat catalase 1 (TdCAT1) exhibits a low CAT activity in vitro, whereas a deleted form of TdCAT1 has better activity compared to the full-length protein (TdCAT460), suggesting that TdCAT1 could present a putative autoinhibitory domain in its C-terminal portion. Moreover, we showed that TdCAT1 positively regulates E. coli cells in response to salt, ionic and osmotic stresses as well as heavy metal stress in solid and liquid mediums. Such effects had not been reported and lead us to suggest that the durum wheat catalase 1 TdCAT1 protein could play a positive role in response to a wide array of abiotic stress conditions.

The Activity of the Durum Wheat (Triticum durum L.) Catalase 1 (TdCAT1) Is Modulated by Calmodulin.

Plant catalases (CAT) are involved in the cellular scavenging of the reactive oxygen species during developmental processes and in response to abiotic and biotic stresses. However, little is known about the regulation of the CAT activity to ensure efficient antioxidant function. Using bioinformatic analyses, we showed that durum wheat catalase 1 (TdCAT1) harbors highly conserved cation-binding and calmodulin binding (CaMBD) domains which are localized at different positions of the protein. As a result, the catalytic activity of TdCAT1 is enhanced in vitro by the divalent cations Mn2+ and Fe2+ and to a lesser extent by Cu2+, Zn2+, and Mg2+. Moreover, the GST-pull down assays performed here revealed that TdCAT1 bind to the wheat CaM (TdCaM1.3) in a Ca2+-independent manner. Furthermore, the TdCaM1.3/Ca2+ complex is stimulated in a CaM-dose-dependent manner by the catalytic activity of TdCAT1, which is further increased in the presence of Mn2+ cations. The catalase activity of TdCAT1 is enhanced by various divalent cations and TdCaM1.3 in a Ca-dependent manner. Such effects are not reported so far and raise a possible role of CaM and cations in the function of CATs during cellular response to oxidative stress.

Recent advances in physiological and molecular mechanisms of heavy metal accumulation in plants.

Among abiotic stress, the toxicity of metals impacts negatively on plants' growth and productivity. This toxicity promotes various perturbations in plants at different levels. To withstand stress, plants involve efficient mechanisms through the implication of various signaling pathways. These pathways enhance the expression of many target genes among them gene coding for metal transporters. Various metal transporters which are localized at the plasma membrane and/or at the tonoplast are crucial in metal stress response. Furthermore, metal detoxification is provided by metal-binding proteins like phytochelatins and metallothioneins. The understanding of the molecular basis of metal toxicities signaling pathways and tolerance mechanisms is crucial for genetic engineering to produce transgenic plants that enhance phytoremediation. This review presents an overview of the recent advances in our understanding of metal stress response. Firstly, we described the effect of metal stress on plants. Then, we highlight the mechanisms involved in metal detoxification and the importance of the regulation in the response to heavy metal stress. Finally, we mentioned the importance of genetic engineering for enhancing the phytoremediation technique. In the end, the response to heavy metal stress is complex and implicates various components. Thus, further studies are needed to better understand the mechanisms involved in response to this abiotic stress.

Functional analysis of TmHKT1;4-A2 promoter through deletion analysis provides new insight into the regulatory mechanism underlying abiotic stress adaptation.

MAIN CONCLUSION: Bioinformatic, molecular, and biochemical analysis were performed to get more insight into the regulatory mechanism by which TmHKT1;4-A2 is regulated. HKT transporters from different plant species have been shown to play important role in plant response to salt. In previous work, TmHKT1;4-A2 gene from Triticum monococcum has been characterized as a major gene for Nax1 QTL (Tounsi et al. Plant Cell Physiol 57:2047-2057, 2016). So far, little is known about its regulatory mechanism. In this study, the promoter region of TmHKT1;4-A2 (1400 bp) was isolated and considered as the full-length promoter (PA2-1400). In silico analysis revealed the presence of important cis-acting elements related to abiotic stresses and phytohormones. Interestingly, our real-time RT-PCR analysis provided evidence that TmHKT1;4-A2 is regulated not only by salt stress but also by osmotic, heavy metal, oxidative, and hormones stresses. In transgenic Arabidopsis plants, TmHKT1;4-A2 is strongly active in vascular tissues of roots and leaves. Through 5'-end deletion analysis, we showed that PA2-1400 promoter is able to drive strong GUS activity under normal conditions and in response to different stresses compared to PA2-824 and PA2-366 promoters. These findings provide new information on the regulatory mechanism of TmHKT1;4-A2 and shed more light on its role under different stresses.

Causal Enzymology and Physiological Aspects May Be Accountable to Membrane Integrity in Response to Salt Stress in Arabidopsis thaliana Lines.

Apart from their significance in the protection against stress conditions, the plant cell membranes are essential for proper development of the diverse surface structures formed on aerial plant organs. In addition, we signal that membrane remodeling and integrity are function of some of causal physiological and enzymological aspects such as the MDA, the ion leakage and also the monitoring of some phytozymes involved in lipid and cellulose metabolisms. Those last ones are related to the membrane structure (lipases and cellulases), that were assessed in durum wheat dehydrin transgenic context (YS, K1-K2, DH2, and DH4), proline metabolic mutant (P5CS1-4) per comparison with the wild-type plant (Wt). We report also the docking data reinforcing the fact that the membrane integrity seems to be function of causal enzymological behaviors, through the molecular dynamic investigation resulting from the dehydrin-phytozyme interactions and also from the inhibition effect of the durum wheat LTP4 on the lipase activity.

Highlight on the expression and the function of a novel MnSOD from diploid wheat (T. monococcum) in response to abiotic stress and heavy metal toxicity.

Superoxide dismutases (SODs) play a pivotal role in improving abiotic stress tolerance in plant cells. A novel manganese superoxide dismutase gene, denoted as TmMnSOD, was identified from Triticum monococcum. The encoded protein displayed high sequence identity with MnSOD family members and was highly homologous to TdMnSOD from durum wheat. Furthermore, the 3D structure analysis revealed that TmMnSOD displayed homotetramer subunit organization, incorporating four Mn2+ ions. Notably, TmMnSOD structure contains predominantly alpha helices with three beta sheets. On the other hand, under stress conditions, TmMnSOD transcript level was significantly up-regulated by salt, oxidative and heavy metal stresses. At the functional level, TmMnSOD imparts tolerance of yeast and E. coli cells under diverse stresses. Promoter analysis of TmMnSOD gene showed the presence of a great number of salt and pathogen-responsive cis-regulatory elements, highlighting the interest of this gene in breeding programs towards improved tolerance to salt stress in wheat.

Localization and expression analysis of a novel catalase from Triticum monococcum TmCAT1 involved in response to different environmental stresses.

Catalase proteins play a crucial role in detoxifying hydrogen peroxide, generated during plant growth, and in response to various environmental stresses. Despite their importance, little is known about their localization and expression in wheat. In this study, we identified and characterized a novel peroxisomal catalase gene from Triticum monococcum, designated as TmCAT1. Phylogenetic analysis revealed that TmCAT1 shared high identity with TdCAT1 and other plant catalases belonging to subfamily 1. We predicted the 3D structure model and the oligomerization arrangement of TmCAT1. Besides, we displayed an arrangement in asymmetric unit, which involved interactions including, mainly, residues from N-terminal domain. Interestingly, sequence analysis indicated that TmCAT1, like TdCAT1, had the peroxisomal targeting signal (PTS1) around its C-terminus. Transient expression of TmCAT1-GFP and TdCAT1-GFP in tobacco leaves revealed that the two fused proteins are targeted into peroxisomes. However, the truncated forms lacking the tripeptide QKL remained in the cytosol. Concerning the expression profile analysis, TmCAT1 is expressed especially in leaves in normal condition. On the other hand, it is up-regulated by different stress incorporating salt, osmotic, oxidative, heavy metal and hormones stresses. Functional analysis by heterologous expression in yeast cells showed that TmCAT1 improved tolerance to multiple abiotic stresses. The presence of important cis-regulatory elements in the promoter region of TmCAT1 strongly reinforces the interest of this gene in plant adaptation to various stresses.

Promoter of the TmHKT1;4-A1 gene of Triticum monococcum directs stress inducible, developmental regulated and organ specific gene expression in transgenic Arbidopsis thaliana.

HKT transporters which mediate Na+-specific transport or Na+-K+ co-transport, play an important role in protecting plants from salinity stress by preventing Na+-over-accumulation in leaves. In this work, a 1508-bp genomic fragment upstream of the TmHKT1;4-A1 translated sequence from Triticum monococcum has been isolated, cloned, and designated as the ''PrTmHKT1;4-A1'' promoter. Sequence analysis of ''PrTmHKT1;4-A1'' revealed the presence of cis-regulatory elements which could be required for abiotic stress and abscisic acid (ABA) responsiveness. The PrTmHKT1;4-A1 sequence was fused to the ß-glucuronidase gene and the resulting construct was transferred into Arabidopsis plants. Histochemical assays of stably transformed Arabidopsis plants showed that PrTmHKT1;4-A1 is active in this heterologous system. Under control conditions, GUS histochemical staining was observed significantly only in leaves of 20-day-old plants. Histological sections prepared at this stage and in leaves revealed activity localized in leaf veins (phloem and bundle sheath). In flowers, GUS activity was detected only in sepals. After 3 days of challenging the plants with salt, dehydration or ABA treatments, the PrTmHKT1;4-A1 transformed Arabidopsis plants showed a substantial increase in the GUS staining in leaves, compared to untransformed plants under the same conditions. Real time qPCR expression analysis of the uidA gene, showed that GusA transcripts were up-regulated by salt, dehydration, and ABA treatments. All together, these results showed that PrTmHKT1;4-A1 is an age-dependent, abiotic-stress-inducible, organ-specific and tissue-specific promoter in a heterologous dicot system.

Salt stress reveals differential physiological, biochemical and molecular responses in T. monococcum and T. durum wheat genotypes.

Salt stress responses implicate a complex mechanism and differ from plant species to another. In this study, we analyzed the physiological, biochemical and molecular responses to salt stress of the diploid wheat (T. monococcum) and compared to the tetraploid wheat (T. durum). Our results showed that the diploid wheat cultivar (cv. Turkey) is relatively tolerant to different salt stress conditions than the tetraploid wheat cultivar (cv. Om Rabia3). This tolerance was manifested by significant germination, plant growth and uptake of water generating cell turgor and development. Moreover, total chlorophyll content was higher in the diploid wheat than that in the tetraploid wheat. The Na+ content in leaf blade of the cv. Om Rabia3 was significantly higher than that of the cv. Turkey, suggesting that the diploid cultivar accumulates less toxic sodium in the photosynthetic tissues. This mechanism could be explained by the recirculation of the toxic ions Na+ into the xylem sap by SOS1 protein, which coordinates with HKT-like proteins to reduce the accumulation of Na+ ions in leaf blade. Interestingly, the expression of the three genes SOS1, HKT and NHX was enhanced under salinity especially in leaf blade of the cv. Turkey. Moreover, this wheat cultivar induced the antioxidative enzymes CAT and SOD activity more efficiently than the other cultivar.

The durum wheat plasma membrane Na+/H+ antiporter SOS1 is involved in oxidative stress response.

We have shown previously that the durum wheat TdSOS1 excludes Na+ and Li+ ions outside cells. Moreover, this protein is activated by Arabidopsis kinase SOS2 through phosphorylation. The elimination of both SOS2 phosphorylation sites and the auto-inhibitory domain produces a hyperactive TdSOS1∆972 form, which have a maximal activity independent from the regulatory SOS2/SOS3 complex. We demonstrated that the expression of TdSOS1 enhances salt tolerance of the transgenic Arabidopsis plants. In this study, we analyzed the response to H2O2-induced oxidative stress of the transgenic Arabidopsis expressing one of the two TdSOS1 forms. Firstly, we showed that the exogenous H2O2 treatment leads to an accumulation of SOS1 transcripts in leaves and roots of the durum wheat and also in the transgenic plants. These transgenic plants showed significant oxidative stress tolerance compared to control plants, especially the plants expressing the hyperactive form. This tolerance was manifested by high proline accumulation and low malonyldialdehyde (MDA), O2˙- and H2O2 contents. Furthermore, the activities of three essential ROS scavenging enzymes (SOD, CAT, and POD) were higher in the transgenic plants under oxidative stress, as compared to control plants. Taken together, these data suggested that TdSOS1 plays a crucial role in response to oxidative stress.

Multiple abiotic stress tolerance of the transformants yeast cells and the transgenic Arabidopsis plants expressing a novel durum wheat catalase.

Catalases are reactive oxygen species scavenging enzymes involved in response to abiotic and biotic stresses. In this study, we described the isolation and functional characterization of a novel catalase from durum wheat, designed TdCAT1. Molecular Phylogeny analyses showed that wheat TdCAT1 exhibited high amino acids sequence identity to other plant catalases. Sequence homology analysis showed that TdCAT1 protein contained the putative calmodulin binding domain and a putative conserved internal peroxisomal targeting signal PTS1 motif around its C-terminus. Predicted three-dimensional structural model revealed the presence of four putative distinct structural regions which are the N-terminal arm, the ß-barrel, the wrapping and the α-helical domains. TdCAT1 protein had the heme pocket that was composed by five essential residues. TdCAT1 gene expression analysis showed that this gene was induced by various abiotic stresses in durum wheat. The expression of TdCAT1 in yeast cells and Arabidopsis plants conferred tolerance to several abiotic stresses. Compared with the non-transformed plants, the transgenic lines maintained their growth and accumulated more proline under stress treatments. Furthermore, the amount of H2O2 was lower in transgenic lines, which was due to the high CAT and POD activities. Taken together, these data provide the evidence for the involvement of durum wheat catalase TdCAT1 in tolerance to multiple abiotic stresses in crop plants.

Bridging Between Proline Structure, Functions, Metabolism, and Involvement in Organism Physiology.

Much is now known about proline multifunctionality and metabolism; some aspects of its biological functions are still unclear. Here, we discuss some cases in the proline, structure, definition, metabolism, compartmentalization, accumulation, plausible functions and also its implication in homeostasis and organism physiology. Indeed, we report the role of proline in cellular homeostasis, including redox balance and energy status and their implication as biocatalyst for aldolase activity. Proline can act as a signaling molecule to modulate mitochondrial functions, influence cell proliferation or cell death, and trigger specific gene expression, which can be essential for plant recovery from stresses. Although, the regulation and the function of proline accumulation, during abiotic stresses, are not yet completely understood. The engineering of proline metabolism could lead to new opportunities to improve plant tolerance against environmental stresses. This atypical amino acid has a potential role in the toxicity during growth of some microorganism, vegetal, and mammalian species. Furthermore, we note that the purpose through the work is to provide a rich, concise, and mostly cohesive source on proline, considered as a platform and an anchor between several disciplines and biological functions.

Durum wheat dehydrin (DHN-5) confers salinity tolerance to transgenic Arabidopsis plants through the regulation of proline metabolism and ROS scavenging system.

MAIN CONCLUSION: The wheat dehydrin (DHN-5) gives birth to salinity tolerance to transgenic Arabidopsis plants by the regulation of proline metabolism and the ROS scavenging system. Dehydrins (DHNs) are involved in plant abiotic stress tolerance. In this study, we reported that salt tolerance of transgenic Arabidopsis plants overexpressing durum wheat dehydrin (DHN-5) was closely related to the activation of the proline metabolism enzyme (P5CS) and some antioxidant biocatalysts. Indeed, DHN-5 improved P5CS activity in the transgenic plants generating a significant proline accumulation. Moreover, salt tolerance of Arabidopsis transgenic plants was accompanied by an excellent activation of antioxidant enzymes like catalase (CAT), superoxide dismutase (SOD) and peroxide dismutase (POD) and generation of a lower level of hydrogen peroxide (H2O2) in leaves compared to the wild-type plants. The enzyme activities were enhanced in these transgenic plants in the presence of exogenous proline. Nevertheless, proline accumulation was slightly reduced in transgenic plants promoting chlorophyll levels. All these results suggest the crucial role of DHN-5 in response to salt stress through the activation of enzymes implicated in proline metabolism and in ROS scavenging enzymes.

Comparative functional analysis of two wheat Na(+)/H (+) antiporter SOS1 promoters in Arabidopsis thaliana under various stress conditions.

The bread wheat TaSOS1 has been previously shown to be induced by salt stress treatment. To further investigate the regulation of the TaSOS1 gene, the two genomic fragments Pr SOS1-AB and Pr SOS1-D have been isolated and sequenced. Pr SOS1-AB and Pr SOS1-D are the promoter regions of SOS1 alleles, which are localised on genomes A and/or B, and on genome D, respectively. Sequence analysis of these two promoters revealed the presence of cis-regulatory elements which could be required for abiotic stress and abscisic acid (ABA) responsiveness. Histochemical assays of stably transformed Arabidopsis T3 plants showed that Pr SOS1-AB and Pr SOS1-D are active in this heterologous system, and their activities were almost the same at early developmental stages (4-, 8- and 12-day-old transgenic Arabidopsis seedlings). Nevertheless, ß-glucuronidase (GUS) activity was detected only in plants carrying the Pr SOS1-AB -gusA construct grown for 20 or 30 days. Furthermore, in these plants, the application of abiotic stress produced an accumulation in gusA transcripts. Taken together, these results show that, in this heterologous dicot system and under normal growth conditions, Pr SOS1-AB and Pr SOS1-D are age-dependent and organ-specific promoters. However, in the presence of different stress conditions, the activities of these two promoters became different and only Pr SOS1-AB is an abiotic stress-inducible promoter at different developmental stages. Thus, Pr SOS1-AB can be used for the development of abiotic stress-tolerant transgenic plants.

A constitutively active form of a durum wheat Na⁺/H⁺ antiporter SOS1 confers high salt tolerance to transgenic Arabidopsis.

The SOS signaling pathway has emerged as a key mechanism in preserving the homeostasis of Na⁺ and K⁺ under saline conditions. We have recently identified and functionally characterized, by complementation studies in yeast, the gene encoding the durum wheat plasma membrane Na⁺/H⁺ antiporter (TdSOS1). To extend these functional studies to the whole plant level, we complemented Arabidopsis sos1-1 mutant with wild-type TdSOS1 or with the hyperactive form TdSOS1∆972 and compared them to the Arabidopsis AtSOS1 protein. The Arabidopsis sos1-1 mutant is hypersensitive to both Na⁺ and Li⁺ ions. Compared with sos1-1 mutant transformed with the empty binary vector, seeds from TdSOS1 or TdSOS1∆972 transgenic plants had better germination under salt stress and more robust seedling growth in agar plates as well as in nutritive solution containing Na⁺ or Li⁺ salts. The root elongation of TdSOS1∆972 transgenic lines was higher than that of Arabidopsis sos1-1 mutant transformed with TdSOS1 or with the endogenous AtSOS1 gene. Under salt stress, TdSOS1∆972 transgenic lines showed greater water retention capacity and retained low Na⁺ and high K⁺ in their shoots and roots. Our data showed that the hyperactive form TdSOS1∆972 conferred a significant ionic stress tolerance to Arabidopsis plants and suggest that selection of hyperactive alleles of the SOS1 transport protein may pave the way for obtaining salt-tolerant crops.

Regulation of durum wheat Na+/H + exchanger TdSOS1 by phosphorylation.

We have identified a plasma membrane Na(+)/H(+) exchanger from durum wheat, designated TdSOS1. Heterologous expression of TdSOS1 in a yeast strain lacking endogenous Na(+) efflux proteins showed complementation of the Na(+)- and Li(+)-sensitive phenotype by a mechanism involving cation efflux. Salt tolerance conferred by TdSOS1 was maximal when co-expressed with the Arabidopsis protein kinase complex SOS2/SOS3. In vitro phosphorylation of TdSOS1 with a hyperactive form of the Arabidopsis SOS2 kinase (T/DSOS2∆308) showed the importance of two essential serine residues at the C-terminal hydrophilic tail (S1126, S1128). Mutation of these two serine residues to alanine decreased the phosphorylation of TdSOS1 by T/DSOS2∆308 and prevented the activation of TdSOS1. In addition, deletion of the C-terminal domain of TdSOS1 encompassing serine residues at position 1126 and 1128 generated a hyperactive form that had maximal sodium exclusion activity independent from the regulatory SOS2/SOS3 complex. These results are consistent with the presence of an auto-inhibitory domain at the C-terminus of TdSOS1 that mediates the activation of TdSOS1 by the protein kinase SOS2. Expression of TdSOS1 mRNA in young seedlings of the durum wheat variety Om Rabia3, using different abiotic stresses (ionic and oxidative stress) at different times of exposure, was monitored by RT-PCR.