Tau Class Research Articles

Walnut JrGSTU23 and JrVHAc4 involve in drought tolerance via JrWRKY2-mediated upstream regulatory pathway

Drought stress negatively affects the walnut productivity. To ensure the comprehensive benefits of walnut, this study aims to explore the molecular mechanism of drought stress response. Therefore, the downstream target genes of JrWRKY2, a drought tolerance regulator of Juglans regia, are identified by analyses of promoter expression, yeast one-hybrid and co-expression. The results showed that JrWRKY2 could bind to the W-BOX like motifs from the promoters of JrGSTU23 (a tau class glutathione S-transferase) and JrVHAc4 (a c subunit of vacuole H+-ATPase). JrGSTU23 and JrVHAc4 could be up-regulated by drought stress and abscisic acid (ABA). The drought tolerance of transgenic Arabidopsis overexpression of JrGSTU23 or JrVHAc4 were promoted significantly compared to wild type (WT) plants; More importantly, the degree of improvement in drought resistance is enhanced by application of ABA. The transcription levels of the genes from signaling pathways of ABA and mitogen-activated protein kinases (MAPKs) in JrGSTU23 and JrVHAc4 overexpression plants were higher than those in WT with different profiles. These results propose a trifurcate feed-forward pathway involving in JrWRKY2, JrGSTU23, and JrVHAc4 functions on drought stress response that associated with ABA and MAPK signaling pathways, which will offer new insights to reveal plant adaptation mechanism to adverse stimulus.

A tau class glutathione-S-transferase (OsGSTU5) confers tolerance against arsenic toxicity in rice by accumulating more arsenic in root

Arsenic (As) considered as one of the hazardous metalloid that hampers various physiological activities in rice. To study the mechanism of As tolerance in rice, one differentially expressed tau class glutathione-S-transferase (OsGSTU5) has been selected and transgenic rice plants with knockdown (KD) and overexpressing (OE) OsGSTU5 were generated. Our results suggested that KD lines became less tolerant to As stress than WT plants, while OE lines showed enhanced tolerance to As. Under As toxicity, OE and KD lines showed enhanced and reduced antioxidant activities such as, SOD, PRX and catalase, respectively indicating its role in ROS homeostasis. In addition, higher malondialdehyde content, poor photosynthetic parameters and higher reactive oxygen species (ROS) in KD plant, suggests that knockdown of OsGSTU5 renders KD plants more susceptible to oxidative damage. Also, the relative expression profile of various transporters such as OsABCC1 (As sequestration), Lsi2 and Lsi6 (As translocaters) and GSH dependent activity of GSTU5 suggests that GSTU5 might help in chelation of As with GSH and sequester it into the root vacuole using OsABCC1 transporter and thus limits the upward translocation of As towards shoot. This study suggests the importance of GSTU5 as a good target to improve the As tolerance in rice

Non-synonymous substitution of evolutionarily conserved residue in Tau class glutathione transferases alters structural and catalytic features

Plant-specific tau glutathione transferases (GSTs) are basically involved in catalysing γ-glutathione (GSH)-dependent conjugation reactions with pesticides and herbicides, which play an important role in the detoxification of pollutants. Given the lack of systematic biochemical and structural information on tau GSTs, the study of their mediated defence mechanisms against toxic compounds has been greatly hindered. Here, we reveal the importance of the Ile residue closely interacting with GSH for the structural stability and catalytic function of GST. Evolutionary conservation analysis indicated that the crucial G-site Ile55 in the SbGSTU6 was converted to Thr53 of SbGSTU7. The comparative biochemical data on SbGSTU6, SbGSTU7 and their mutants showed that the substitution of Ile by Thr caused significant decrease in the affinity and catalytic efficiency of the GSTs. The unfavourable structural flexibility and pKa distribution of the active cavity residues were also demonstrated. Crystallography studies and molecular dynamics simulations showed that the conversion resulted in the hydrogen bond recombination with GSH and conformational rearrangement of GST active cavity, in which the Ile residue was more conducive to the formation of enzyme substrate complexes. The extensive biochemical and structural data not only reveal the critical role of the conserved G-site Ile residue in catalysing GSH-conjugate reactions but also provide valuable resources for the development of GST engineering in analytical and agricultural biotechnology.

Genome-wide analysis of the GlutathioneS-Transferase family in wild Medicago ruthenica and drought-tolerant breeding application ofMruGSTU39 gene in cultivated alfalfa.

Transformation of MruGSTU39 in M. ruthenica and alfalfa enhanced growth and survival of transgenic plants by up-regulating GST and glutathione peroxidase activity to detoxify ROS under drought stress. Glutathione S-transferases (GSTs) are ubiquitous supergenefamily which play crucial roles in detoxification of reactive oxygenspecies (ROS). Despite studies on GSTs, few studies have focused on them in perennial, wild plant species with high tolerance to environmental stress. Here, we identified 66 MruGST genes from the genome of Medicago ruthenica, a perennial legume species native to temperate grasslands with high tolerance to environmental stress. These genes were divided into eight classes based on their conserved domains, phylogenetic tree and gene structure, with the tau class being the most numerous. Duplication analysis revealed that GST family in M. ruthenica was expanded by segmental and tandem duplication. Several drought-responsive MruGSTs were identified by transcriptomic analyses. Of them, expression of MruGSTU39 was up-regulated much more in a tolerant accession by drought stress. Transformation of MruGSTU39 in M. ruthenica and alfalfa (Medicago sativa) enhanced growth and survival of transgenic seedlings than their wild-type counterparts under drought. We demonstrated that MruGSTU39 can detoxify ROS to reduce its damage to membrane by up-regulating activities of GST and glutathione peroxidase. Our findings provide full-scaleknowledge on GST family in the wild legume M. ruthenica with high tolerance to drought, and highlight improvement tolerance of legume forages to drought using genomic information of M. ruthenica.

Identification of Glutathione S-Transferase Genes in Hami Melon (Cucumis melo var. saccharinus) and Their Expression Analysis Under Cold Stress.

As a group of multifunctional enzymes, glutathione S-transferases (GSTs) participate in oxidative stress resistance and cellular detoxification. Here, we identified 39 CmGST genes with typical binding sites from the Hami melon genome, and they can be classified into seven subfamilies. Their molecular information, chromosomal locations, phylogenetic relationships, synteny relationships, gene structures, protein–protein interactions, structure of 3-D models, and expression levels under cold stress were analyzed. Expression analysis indicates that cold-tolerant Jia Shi-310 (JS) had higher GST enzyme activities and expression levels of 28 stress-related genes under cold stress. Some CmGSTs belonging to Tau, Phi, and DHAR classes play significant roles under cold stress, and they could be regarded as candidate genes for further studies. The present study systematically investigated the characterization of the Hami melon GST gene family, extending our understanding of Hami melon GST mediated stress-response mechanisms in this worldwide fruit.

Directed Evolution of a Glutathione Transferase for the Development of a Biosensor for Alachlor Determination

In the present work, DNA recombination of three homologous tau class glutathione transferases (GSTUs) allowed the creation of a library of tau class GmGSTUs. The library was activity screened for the identification of glutathione transferase (GST) variants with enhanced catalytic activity towards the herbicide alachlor (2-chloro-2′,6′-diethyl-N-(methoxymethyl)acetanilide). One enzyme variant (GmGSTsf) with improved catalytic activity and binding affinity for alachlor was identified and explored for the development of an optical biosensor for alachlor determination. Kinetics analysis and molecular modeling studies revealed a key mutation (Ile69Val) at the subunit interface (helix α3) that appeared to be responsible for the altered catalytic properties. The enzyme was immobilized directly on polyvinylidenefluoride membrane by crosslinking with glutaraldehyde and was placed on the inner surface of a plastic cuvette. The rate of pH changes observed as a result of the enzyme reaction was followed optometrically using a pH indicator. A calibration curve indicated that the linear concentration range for alachlor was 30–300 μM. The approach used in the present study can provide tools for the generation of novel enzymes for eco-efficient and environment-friendly analytical technologies. In addition, the outcome of this study gives an example for harnessing protein symmetry for enzyme design.

Comparative Analysis of the Glutathione S-Transferase Gene Family of Four Triticeae Species and Transcriptome Analysis of GST Genes in Common Wheat Responding to Salt Stress.

Glutathione S-transferases (GSTs) are ancient proteins encoded by a large gene family in plants, which play multiple roles in plant growth and development. However, there has been little study on the GST genes of common wheat (Triticum aestivum) and its relatives (Triticum durum, Triticum urartu, and Aegilops tauschii), which are four important species of Triticeae. Here, a genome-wide comprehensive analysis of this gene family was performed on the genomes of common wheat and its relatives. A total of 346 GST genes in T. aestivum, 226 in T. durum, 104 in T. urartu, and 105 in Ae. tauschii were identified, and all members were divided into ten classes. Transcriptome analysis was used to identify GST genes that respond to salt stress in common wheat, which revealed that the reaction of GST genes is not sensitive to low and moderate salt concentrations but is sensitive to severe concentrations of the stressor, and the GST genes related to salt stress mainly come from the Tau and Phi classes. Six GST genes which respond to different salt concentrations were selected and validated by a qRT-PCR assay. These findings will not only provide helpful information about the function of GST genes in Triticeae species but also offer insights for the future application of salt stress resistance breeding in common wheat.

Effects of conserved Arg20, Glu74 and Asp77 on the structure and function of a tau class glutathione S-transferase in rice.

The relative position of domains is critical for enzymatic properties of tau class glutathione S-transferases, and altering the position of linker far away from the active center affects catalytic property. Glutathione S-transferases (GSTs) are a family of phase II detoxification enzymes whose main function is to improve plant resistance to stresses. To understand the structural effects of tau class GSTs on their function, using OsGSTU17 as an example, we predicted the residues involved in the interactions between its domains and linker region. We further detected the structural changes in mutants and the corresponding changes in terms of substrate activity and kinetic parameters. Four pairs of residues, including Ala14 and Trp165, Arg20 and Tyr154, Glu74 and Arg98, Asp77 and Met87, forming hydrogen bonds and salt bridges were found to play important roles in maintaining the relative position between the domains and linker region inside the protein. The hydrogen bond between Trp165 and Ala14 affected the structural stability has been demonstrated in our previous study. The mutant R20A lost almost all catalytic activity. Interestingly, the mutant E74A exhibited a significant decrease in activity towards 7-chloro-4-nitrobenzo-2-oxa-1, 3-diazole, 1-chloro-2, 4-dinitrobenzene and 4-nitrobenzyl chloride, while its activity towards substrate cumene hydroperoxide remained unchanged. Compared with other mutants, the mutant D77A exhibited decreased affinity to its substrates and increased activity towards 1-chloro-2, 4-dinitrobenzene and cumene hydroperoxide, but its thermodynamic stability did not change significantly. The relative position of individual domain was critical for enzymatic properties, and the linker which is far away from the active site could change the enzymatic properties of GSTs via altering the relative position of the individual domain. Our results provide insights into the relationship between structure and function of tau class GSTs.

OsGSTU6 Contributes to Cadmium Stress Tolerance in Rice by Involving in Intracellular ROS Homeostasis

Glutathione S-transferases (GSTs) are multifunctional proteins that play important roles in cellular detoxification as well as in plant growth and development. Here, we cloned a new rice tau class GST gene, OsGSTU6, and evaluated its roles in regulating cadmium (Cd) stress tolerance of rice. OsGSTU6 is located both in the cytosol and nucleus of cells. The transcripts of OsGSTU6 were detected in the whole life cycle of rice with higher levels in the leaf blade and sheath at heading stage. The transcripts of OsGSTU6 were significantly stimulated by a number of environmental factors including several phytohormones, abiotic stresses, and heavy metals, suggesting its stress-responsive characteristics. Moreover, the overexpression (OE) of OsGSTU6 in rice reduced the accumulation of Cd in leaf and enhanced the tolerance of the plant to Cd stress, while knockdown (RNAi) of OsGSTU6 triggered Cd accumulation in rice leaves and decreased stress tolerance. Meanwhile, the expression levels of several candidate genes responsible for scavenging reactive oxygen species (ROS) were downregulated in OE of OsGSTU6 lines. In addition, the content of superoxide anions (O2−) and glutathione showed reduction in OE plants, but was augmented in RNAi plants under both normal growth and Cd stress, compared with wild type. Furthermore, both the yeast two-hybrid assay and firefly luciferase complementation imaging (LCI) assay demonstrated that OsGSTU6 might function with dimers in plant cells. All these results suggest that OsGSTU6 may play an important role in Cd stress tolerance of rice by involving in the intracellular ROS homeostasis.

Genome-wide analysis and transcript profiling identify several abiotic and biotic stress-responsive Glutathione S-transferase genes in soybean

Abiotic and biotic stresses stimulate the generation of free radicals and reactive oxygen species, which results in oxidative stress. Glutathione-S-transferase (GST) is a large class of multifunctional enzymes that exists widely in plants. The addition of reduced glutathione by GSTs protects the cell from various toxic molecules and thus provide tolerance against adverse conditions. In this study, we have performed a genome-wide identification, characterization and expression analysis of GST gene family in soybean (Glycine max) to know their function. We have identified the presence of at least 126 putative GST genes in soybean. Among them, presence of a total of 33 pairs of duplicated genes might contribute towards the expansion of GST family members in soybean. Based on the phylogenetic analysis, GmGSTs could be further subdivided into 12 classes. GmGST tau class possessed the maximum number of 68 members, followed by phi (14 members) and lambda (7 members) classes. Expression of all 126 GmGST transcripts was analyzed in 26 anatomical tissues and 7 developmental stages. Moreover, expression profiling was expanded in response to various abiotic and biotic stresses. Four genes (GmGSTU63, GmGSTU73, GmGSTF2, and GmGSTT5) were found to be highly upregulated in response to four abiotic stresses- dehydration, drought, temperature and ozone; and two biotic stress conditions. This comprehensive genome-wide analysis and expression profiling will help to understand the function of GST in adverse environmental circumstances as well as enhancing the genetic modification of soybean.

The AtGSTU7 gene influences glutathione-dependent seed germination under ABA and osmotic stress in Arabidopsis

Glutathione S-transferases (GSTs) play important roles in metabolism and detoxification of plant cells. However, their functions during development are less well understood. Arabidopsis AtGSTU7 (AT2G29420) encodes a Tau class GST. Here we provide the AtGSTU7 was abundantly expressed in seeds and in roots at an early stage of germination. AtGSTU7 expression was repressed by exogenous ABA and promoted by osmotic stress. A null mutant of AtGSTU7 (atgstu7) accumulated higher contents of reduced GSH and decreased amounts of endogenous H2O2 in seedlings. The atgstu7 plants showed decreased osmotic tolerance during seed germination, which was influenced by GSH and ABI3 gene expression. The results suggested that AtGSTU7 involvement in seed germination is mediated by GSH–ROS homeostasis and ABA signaling.

Genome-wide analysis of glutathione S-transferase gene family in G. max

Glutathione S-transferases (GSTs) are prevalent enzymes, capable of multifaceted tasks like cellular detoxification and stress resistance. Our study was focused on the thorough genome-wide investigation of the GST gene family in the soybean (G. max). The results of this study led to the identification of no less than 74 GST genes in soybean, which is higher than several other reported species. Phylogenetic analysis indicated that GSTs can be subdivided into various classes with each individual class being significantly conserved. Maximum members of the GST family has found in tau class being 37 members, preceding phi and lambda class. Studies for chromosomal localization demonstrated significantly lopsided allocation of GmGST genes all over the soybean genome. Several members of GmGST revealed tissue-specific expression during the transcript profiling for 74 GmGST genes. Furthermore, responsive cis-regulatory elements have found in GmGST genes during promoter analysis. The 12 paralogous pairs have found due to segmental duplication in the GmGST gene family and present on 19 different chromosomes. The Ka/Ks ratio of paralogous pair showed that the evolution in duplicated genes was under purifying selection and duplication has done during recent and ancient times as well. Such findings will help for future studies on GST genes in soybean related to the functional and evolutionary characterization.

Genome-wide analysis of glutathione S-transferase gene family in chickpea suggests its role during seed development and abiotic stress.

Glutathione S-transferases (GSTs) are multifunctional proteins that help in oxidative stress metabolism and detoxification of xenobiotic compounds. Studies pertaining to GST gene family have been undertaken in various plant species, however no information is available with respect to GST genes in chickpea. In the current study, we identified a total of 51 GST encoding genes in chickpea (CaGST) genome. Phylogenetic analysis revealed that GST gene family can be divided into eleven distinct classes. Tau and phi were the major classes in chickpea and one third of the CaGST genes represented segmental duplication and purifying selection was common among these genes. Expression of many CaGST genes, in particular, members of tau class were found to be upregulated under abiotic stress conditions. In addition, CaGST genes displayed differential expression patterns across diverse organs/tissues, suggesting their roles in developmental processes. Many CaGST genes showed opposite expression pattern in small- and large-seeded chickpea cultivars during seed development. Higher expression of CaGST genes in small-seeded cultivar at maturation stages of seed development suggested their important role in seed development and seed size/weight determination in chickpea. Overall, these results provide a comprehensive information on GST gene family members in chickpea and is expected to provide a rational platform to explore versatile role of these genes in semi-arid legume crops.

Identification and characterization of the glutathione S-Transferase (GST) family in radish reveals a likely role in anthocyanin biosynthesis and heavy metal stress tolerance.

Glutathione S-transferases (GSTs) are a large complex family of enzymes (EC 2.5.1.18) that play vital roles in flavonoid metabolism and plant growth and development and are responsive to heavy metal stress. However, knowledge about GST genes in radish (a vegetable crop with an extraordinary capacity to adapt to heavy metal stresses) is limited. Therefore, it is critical to identify putative candidate GST genes responsible for heavy metal stress tolerance and anthocyanin biosynthesis. In this study, we first identified 82 R. sativus GST (RsGST) genes using various bioinformatic approaches, and their expression profiles were characterized from RNAseq data. These RsGST genes could be grouped into 7 major subclasses: tau (43 members), phi (21 members), tetrachlorohydroquinone dehalogenase (7 members), dehydroascorbat reductase (5 members), zeta (3 members), lambda (2 members) and theta (1 member). In addition, most of the RsGST genes showed organ-specific expression in our study. Moreover, the transcripts of RsGSTF12-1 and RsGSTF12-2, belonging to the phi class, might be candidates encoding anthocyanin transporters in carmine radish, whereas the tau class, consisting of RsGSTU13-1, RsGSTU19, RsGSTU24-1, and RsGSTU3, and theta class, consisting of RsGSTT1-1, might be defend radish against adverse heavy metal stresses. These results will aid in understanding the functions of the GST family related to heavy metal stress and anthocyanin biosynthesis, thereby potentially improving radish breeding programs for high-pigment-content material as well as HM-tolerant material.

Molecular cloning and biochemical characterization of a Tau class glutathione S-transferase from Pinus brutia Ten

KEY MESSAGE: A new Tau class GST gene was cloned from Pinus brutia Ten. cDNA sequence was analysed for conserved sequences. Substrate specificity, optimum pH, and temperature values of the recombinant PbGST Tau enzyme were determined. Tau class glutathione S-transferases (GSTs) are essential enzymes for detoxification in plants. To date, a lot of the members of this family have been characterized from different plants but the studies on the conifers are very scarce. This study investigates for the first time molecular cloning and biochemical characterization of a Tau class GST gene (PbGST Tau) from Pinus brutia Ten. The full length PbGST Tau ORF was 687 bp having a molecular mass of 27.37 kDa. Catalytic and ligand binding sites of PbGST Tau are well conserved and shared maximum identity with Pinus tabulaeformis GST Tau. Kinetic analysis with respect to 1-chloro-2,4-dinitrobenzene (CDNB) and ethacrynic acid (ECA) as substrates exhibited a Kₘ of 3.66 mM and 0.3 mM, respectively. PbGST Tau enzyme had an optimum activity at pH 6.0 and 8.0 when CDNB and ECA were used as substrate, respectively. The highest activity was measured at 25 °C. Through enzyme assays, phylogenetic analysis and structural modelling, we provide a detailed characterization of the PbGST Tau gene and the enzyme. This study is going to provide new insights into the phylogenetic and biochemical analysis of GST family in conifers.

Possible Metsulfuron Herbicide Detoxification by a Oryza sativa L. Glutathione S-transferase Enzyme.

Abstract Rice (Oryza sativa L.) is one of the most important crops in the world, and it is considered the primary source of nutritional layout in developing countries in Asia. The glutathione S-transferases (GSTs) superfamily confers to rice protection against biotic and abiotic stress, and herbicide resistance. However, the three-dimensional structure of a GST Tau class, is unsolved. The objectives of this work were to develop a reliable comparative model for the s-transferase glutathione class Tau 4 from rice, and simulate docking interactions, against herbicides bentazon and metsulfuron. Results showed that the predicted model is reliable and has structural quality. Ramachandran plot set 91.9% of the residues in the most favored regions. All complexes showed negative binding energies values; and metsulfuron docked to the glutathione tripeptide, and it represents a possible insilico evidence of glutathione conjugation with this herbicide.

Effects of Substrate-Binding Site Residues on the Biochemical Properties of a Tau Class Glutathione S-Transferase from Oryza sativa.

Glutathione S-transferases (GSTs)—an especially plant-specific tau class of GSTs—are key enzymes involved in biotic and abiotic stress responses. To improve the stress resistance of crops via the genetic modification of GSTs, we predicted the amino acids present in the GSH binding site (G-site) and hydrophobic substrate-binding site (H-site) of OsGSTU17, a tau class GST in rice. We then examined the enzyme activity, substrate specificity, enzyme kinetics and thermodynamic stability of the mutant enzymes. Our results showed that the hydrogen bonds between Lys42, Val56, Glu68, and Ser69 of the G-site and glutathione were essential for enzyme activity and thermal stability. The hydrophobic side chains of amino acids of the H-site contributed to enzyme activity toward 4-nitrobenzyl chloride but had an inhibitory effect on enzyme activity toward 1-chloro-2,4-dinitrobenzene and cumene hydroperoxide. Different amino acids of the H-site had different effects on enzyme activity toward a different substrate, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole. Moreover, Leu112 and Phe162 were found to inhibit the catalytic efficiency of OsGSTU17 to 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, while Pro16, Leu112, and Trp165 contributed to structural stability. The results of this research enhance the understanding of the relationship between the structure and function of tau class GSTs to improve the abiotic stress resistance of crops.

Regulation of boron toxicity responses via glutathione-dependent detoxification pathways at biochemical and molecular levels in Arabidopsisthaliana

The fine-tuned regulation of the Halliwell-Asada cycle (ascorbate-glutathione pathway) in Arabidopsis thaliana under boron (B) toxicity was shown in our previous report. In this study, we investigated the expression levels of some members of the glutathione S-transferase (GST) superfamily, such as phi (GSTF2, GSTF6, GSTF7, and GSTF8), tau (GSTU19), and zeta (GSTZ1) classes in Arabidopsis thaliana that were exposed to 1 mM boric acid (1B) and 3 mM boric acid (3B). Additionally, the expression levels of genes for glutathione (GSH) and phytochelatin biosynthesis as well as miR169 and miR156 were evaluated in Arabidopsis thaliana exposed to 1B and 3B. Moreover, changes in the levels of total GST activity; GSH; and total, protein-bound, and nonprotein thiols were spectrophotometrically determined. GSH levels and nonprotein thiol content did not change significantly following both B-toxicity conditions. Expression levels of GSH1 and GSH2 stayed stable under 1B toxicity; however, GSH1 expression increased significantly under 3B conditions in Arabidopsis thaliana. The expression levels of four genes from phi class members of GST were not dramatically changed under B-toxicity conditions. However, the transcript levels of miR169, ATGSTU19, and ATGSTZ1 were significantly increased after 1B and 3B exposure. These GST genes may have a role in the dramatic increase of total GST activity under toxic B. To the best of our knowledge, this is the first report displaying an integrative view of high-B-induced regulation of GSH-dependent enzymatic machinery at different biological organization levels in Arabidopsis thaliana.

Detoxification of Reactive Carbonyl Species by Glutathione Transferase Tau Isozymes.

Oxidative stimuli to living cells results in the formation of lipid peroxides, from which various aldehydes and ketones (oxylipin carbonyls) are inevitably produced. Among the oxylipin carbonyls, those with an α,β-unsaturated bond are designated as reactive carbonyl species (RCS) because they have high electrophilicity and biological activity. Plants have arrays of dehydrogenases and reductases to metabolize a variety of RCS that occur in the cells, but these enzymes are not efficient to scavenge the most toxic RCS (i.e., acrolein) because they have only low affinity. Two glutathione transferase (GST) isozymes belonging to the plant-specific Tau class were recently observed to scavenge acrolein with KM values at a submillimolar level. This suggests that GST could also be involved in the defense system against RCS. We tested the activities of 23 Tau isozymes of Arabidopsis thaliana for five types of RCS, and the results revealed that 11 isozymes recognized either acrolein or 4-hydroxy-(E)-2-nonenal or both as a substrate(s). Such RCS-scavenging activities indicate the potential contribution of GST to RCS scavenging in plants, and they may account for the stress tolerance conferred by several Tau isozymes. RCS are therefore a strong candidate for endogenous substrates of plant GSTs.

The Arabidopsis glutathione transferases, AtGSTF8 and AtGSTU19 are involved in the maintenance of root redox homeostasis affecting meristem size and salt stress sensitivity

The tau (U) and phi (F) classes of glutathione transferase (GST) enzymes reduce the glutathione (GSH) pool using GSH as a co-substrate, thus influence numerous redox-dependent processes including hormonal and stress responses. We performed detailed analysis of the redox potential and reactive oxygen species levels in longitudinal zones of 7-day-old roots of Arabidopsis thaliana L. Col-0 wild type and Atsgtf8 and Atgstu19 insertional mutants. Using redox-sensitive cytosolic green fluorescent protein (roGFP2) the redox status of the meristematic, transition, and elongation zones was determined under control and salt stress (3-hour of 75 or 150 mM NaCl treatment) conditions. The Atgstu19 mutant had the most oxidized redox status in all root zones throughout the experiments. Using fluorescent dyes significantly higher superoxide radical (O2−) levels was detected in both Atgst mutants than in the Col-0 control. Salt treatment resulted in the highest O2− increase in the Atgstf8 root, while the amount of H2O2 elevated most in the case of Atgstu19. Moreover, vitality decreased in Atgstu19 roots more than in wild type under salt stress. Our results indicate that AtGSTF8 and especially the AtGSTU19 proteins function in the root fine-tuning the redox homeostasis both under control and salt stress conditions.