O-acetylserine Thiol Lyase Research Articles

Biofortifying multiple micronutrients and decreasing arsenic accumulation in rice grain simultaneously by expressing a mutant allele of OAS-TL gene.

Rice grains typically contain relatively high levels of toxic arsenic (As) but low levels of essential micronutrients. Biofortification of essential micronutrients while decreasing As accumulation in rice would benefit human nutrition and health. We generated transgenic rice expressing a gain-of-function mutant allele astol1 driven by the OsGPX1 promoter. astol1 encodes a plastid-localized O-acetylserine (thiol) lyase (OAS-TL) with Ser189Asn substitution (OsASTOL1S189N), which enhances cysteine biosynthesis by forming an indissociable cysteine synthase complex with its partner serine acetyltransferase (SAT). The effects on growth, As tolerance, and nutrient and As accumulation in rice grain were evaluated in hydroponic, pot and field experiments. The expression of OsASTOL1S189N in pOsGPX1::astol1 transgenic lines enhanced SAT activity, sulphate uptake, biosynthesis of cysteine, glutathione, phytochelatins and nicotianamine, and enhanced tolerance to As. The expression of OsASTOL1S189N decreased As accumulation while increased the accumulation of multiple macronutrients (especially sulphur, nitrogen and potassium) and micronutrients (especially zinc and selenium) in rice grain in a pot experiment and two field experiments, and had little effect on plant growth and grain yield. Our study provides a new strategy to genetically engineer rice to biofortify multiple essential nutrients, reducing As accumulation in rice grain and enhancing As tolerance simultaneously.

Effects of Nitrogen Deficiency on the Photosynthesis, Chlorophyll a Fluorescence, Antioxidant System, and Sulfur Compounds in Oryza sativa.

Decreasing nitrogen (N) supply affected the normal growth of Oryza sativa (O. sativa) seedlings, reducing CO2 assimilation, stomatal conductance (gs), the contents of chlorophylls (Chl) and the ratio of Chl a/Chl b, but increasing the intercellular CO2 concentration. Polyphasic chlorophyll a fluorescence transient and relative fluorescence parameters (JIP test) results indicated that N deficiency increased Fo, but decreased the maximum quantum yield of primary photochemistry (Fv/Fm) and the maximum of the IPphase, implying that N-limiting condition impaired the whole photo electron transport chain from the donor side of photosystem II (PSII) to the end acceptor side of PSI in O. sativa. N deficiency enhanced the activities of the antioxidant enzymes, such as ascorbate peroxidase (APX), guaiacol peroxidase (GuPX), dehydro-ascorbate reductase (DHAR), superoxide dismutase (SOD), glutathione peroxidase (GlPX), glutathione reductase (GR), glutathione S-transferase (GST) and O-acetylserine (thiol) lyase (OASTL), and the contents of antioxidant compounds including reduced glutathione (GSH), total glutathione (GSH+GSSG) and non-protein thiol compounds in O. sativa leaves. In contrast, the enhanced activities of catalase (CAT), DHAR, GR, GST and OASTL, the enhanced ASC-GSH cycle and content of sulfur-containing compounds might provide protective roles against oxidative stress in O. sativa roots under N-limiting conditions. Quantitative real-time PCR (qRT-PCR) analysis indicated that 70% of the enzymes have a consistence between the gene expression pattern and the dynamic of enzyme activity in O. sativa leaves under different N supplies, whereas only 60% of the enzymes have a consistence in O. sativa roots. Our results suggested that the antioxidant system and sulfur metabolism take part in the response of N limiting condition in O. sativa, and this response was different between leaves and roots. Future work should focus on the responsive mechanisms underlying the metabolism of sulfur-containing compounds in O. sativa under nutrient deficient especially N-limiting conditions.

S-Benzyl-L-cysteine Inhibits Growth and Photosynthesis, and Triggers Oxidative Stress in Ipomoea grandifolia

L-cysteine, a precursor of essential components for plant growth, is synthesized by the cysteine synthase complex, which includes O-acetylserine(thiol) lyase (OAS-TL) and serine acetyltransferase. In this work, we investigated how S-benzyl-L-cysteine (SBC), an OAS-TL inhibitor, affects the growth, photosynthesis, and oxidative stress of Ipomoea grandifolia plants. SBC impaired gas exchange and chlorophyll a fluorescence, indicating damage that compromised photosynthesis and reduced plant growth. Critical parameters such as the electron transport rate (J), triose phosphate utilization (TPU), light-saturation point (LSP), maximum carboxylation rate of Rubisco (Vcmax), and light-saturated net photosynthetic rate (PNmax) decreased by 19%, 20%, 22%, 23%, and 24%, respectively. The photochemical quenching coefficient (qP), quantum yield of photosystem II photochemistry (ϕPSII), electron transport rate through PSII (ETR), and stomatal conductance (gs) decreased by 12%, 19%, 19%, and 34%, respectively. Additionally, SBC decreased the maximum fluorescence yield (Fm), variable fluorescence (Fv), and chlorophyll (SPAD index) by 14%, 15%, and 15%, respectively, indicating possible damage to the photosynthetic apparatus. SBC triggered root oxidative stress by increasing malondialdehyde, reactive oxygen species, and conjugated dienes by 30%, 55%, and 61%, respectively. We hypothesize that dysfunctions in sulfur-containing components of the photosynthetic electron transport chain, such as the cytochrome b6f complex, ferredoxin, and the iron–sulfur (Fe-S) centers are the cause of these effects, which ultimately reduce the efficiency of electron transport and hinder photosynthesis in I. grandifolia plants. In short, our findings suggest that targeting OAS-TL with inhibitors like SBC could be a promising strategy for the development of novel herbicides.

Gene identification, expression analysis, and molecular docking of SAT and OASTL in the metabolic pathway of selenium in Cardamine hupingshanensis

Key messageIdentification of selenium stress-responsive expression and molecular docking of serine acetyltransferase (SAT) and O-acetyl serine (thiol) lyase (OASTL) in Cardamine hupingshanensis.A complex coupled with serine acetyltransferase (SAT) and O-acetyl serine (thiol) lyase (OASTL) is the key enzyme that catalyzes selenocysteine (Sec) synthesis in plants. The functions of SAT and OASTL genes were identified in some plants, but it is still unclear whether SAT and OASTL are involved in the selenium metabolic pathway in Cardamine hupingshanensis. In this study, genome-wide identification and comparative analysis of ChSATs and ChOASTLs were performed. The eight ChSAT genes were divided into three branches, and the thirteen ChOASTL genes were divided into four branches by phylogenetic analysis and sequence alignment, indicating the evolutionary conservation of the gene structure and its association with other plant species. qRT-PCR analysis showed that the ChSAT and ChOASTL genes were differentially expressed in different tissues under various selenium levels, suggesting their important roles in Sec synthesis. The ChSAT1;2 and ChOASTLA1;2 were silenced by the VIGS system to investigate their involvement in selenium metabolites in C. hupingshanensis. The findings contribute to understanding the gene functions of ChSATs and ChOASTLs in the selenium stress and provide a reference for further exploration of the selenium metabolic pathway in plants.

Mitigation of lead toxicity in Brassica juncea L. by sulphur application – Via various biochemical and transcriptomic strategies

The present study was designed to elucidate whether exogenously applied sulphur (S) (S1; 100 ppm, S2; 200 ppm S) alleviate lead (Pb)-induced (Pb1; 100 ppm, Pb2; 200 ppm and Pb3; 300 ppm Pb) stress in the leaves of Brassica juncea. To study this effect, oxidative stress biomarkers (hydrogen peroxide and malondialdehyde), enzymatic antioxidants (ascorbate peroxidase (APX), glutathione reductase (GR)), non-enzymatic antioxidants (chlorophyll and carotenoids) and S-metabolic enzymes like adenosine triphosphate sulfurylase (ATPS), O-acetylserine(thiol)lyase (OASTL) were studied. Pb treatment dramatically inhibited plant development by increasing lipid peroxidation and hydrogen peroxide (H2O2) accumulation by 34.82 and 238.89 % respectively at 90 days plant. The overproduced oxidative biomarkers alter plant cell homeostasis. In contrast, APX (25.56 %) and GR (32.45 %) activities, chlorophyll (15.51 %) and carotenoids (30 %) contents increased under similar Pb treatments and regulated ROS-antioxidant balance. Exogenous S application, maintained the redox status of cell by further increasing the enzymatic activities. Pbtreatment increased S assimilation in Brassicaby elevating enzyme activities of ATPS andOASTL, which was further augmented by S supplementation toPb-stressed plants. Besides gene expression of BjSULT, BjOASTL and BjGR1 exhibited slight increase under Pb stress, S application to the stressed plants doubled the expression and assisted the stress resistance. Overall, our findings demonstrate that S protect Pb-stressed Brassica seedlings, implying that S could be an ideal choicefor reducing Pb toxicity in crops.

Phenylalanine-cultured Meyerozyma caribbica enhances cysteine metabolism to improve black spot disease resistance in jujube

The yeast strain Meyerozyma caribbica (M. caribbica) is a highly effective control measure for black spot rot in jujube. Our previous studies have shown that phenylalanine can improve the biocontrol efficacy of M. caribbica, but the related molecular mechanism is unknown. This study pointed out that phenylalanine could improve the ability of M. caribbica to induce black spot rot resistance in jujube fruit. Transcriptomic results showed the expression of several amino acid metabolism genes were up-regulated after M. caribbica treatment, and these genes was mainly enriched in the cysteine metabolism pathway. Phenylalanine-cultured M. caribbica enhanced the activities of O-acetylserine (thiol)lyase (OASTL), L-cysteine desulfhydrase (LCD), and D-cysteine desulfhydrase (DCD) in the cysteine metabolism pathway of jujube fruit, promoted the accumulation of cysteine, hydrogen sulfide, and glutathione, and increased the expression of CysE, CysK, LCD, BCAT, and AGXT. In addition, it stimulated the creation of hydrogen peroxide and improved SOD, CAT, and POD activities in jujube. In summary, phenylalanine-cultured M. caribbica improved jujube fruit black spot resistance by activating cysteine metabolism.

Nitric oxide and AMF-mediated regulation of soil enzymes activities, cysteine-H2S system and thiol metabolites in mitigating chromium (Cr (VI)) toxicity in pigeonpea genotypes.

Cr (VI) hampers plant growth and yield by reducing essential nutrient uptake as it competes for phosphate and sulfate transporters. Nitric oxide (NO) and mycorrhization play important roles in mitigating Cr (VI) toxicity. Present study aimed to compare the potential of AMF (Arbuscular mycorrhizal fungi)-Rhizoglomus intraradices and NO (0.25mM) in alleviating Cr (VI) stress (0, 10 and 20mg/kg) in two differentially tolerant pigeonpea genotypes (Pusa 2001 and AL 201). Cr (VI) toxicity reduced growth, mycorrhizal colonization, nutrient uptake, and overall productivity by inducing reactive oxygen species (ROS) generation, with AL 201 more sensitive than Pusa 2001. NO and AM enhanced activities of soil enzymes, thereby increasing nutrients availability as well as their uptake, with AM more effective than NO. Both amendments reduced oxidative stress and restricted Cr (VI) uptake by increasing the activities of antioxidant and S- assimilatory enzymes, with Pusa 2001 more responsive than AL 201. NO was relatively more efficient in regulating cysteine-H2S system by increasing the activities of biosynthetic enzymes (ATP-sulfurylase (ATPS), O-acetylserine thiol lyase (OASTL), D-cysteine desulfhydrase (DCD) and L-cysteine desulfhydrase (LCD), while AM significantly increased glutathione reductase (GR), γ-glutamylcysteine synthetase (γ-ECS) enzymes activities and resultant glutathione (GSH), phytochelatins (PCs), and non-protein thiols (NP-SH) synthesis. Moreover, co-application of NO and AM proved to be highly beneficial in negating the toxic effects of Cr (VI) due to functional complementarity between them. Study suggested the combined use of NO and AM as a useful strategy in re-establishing pigeonpea plants growing in Cr (VI)-stressed environments.

Natural Variation in OASC Gene for Mitochondrial O-Acetylserine Thiollyase Affects Sulfate Levels in Arabidopsis.

Sulfur plays a vital role in the primary and secondary metabolism of plants, and carries an important function in a large number of different compounds. Despite this importance, compared to other mineral nutrients, relatively little is known about sulfur sensing and signalling, as well as about the mechanisms controlling sulfur metabolism and homeostasis. Sulfur contents in plants vary largely not only among different species, but also among accessions of the same species. We previously used associative transcriptomics to identify several genes potentially controlling variation in sulfate content in the leaves of Brassica napus, including an OASC gene for mitochondrial O-acetylserine thiollyase (OAS-TL), an enzyme involved in cysteine synthesis. Here, we show that loss of OASC in Arabidopsis thaliana lowers not only sulfate, but also glutathione levels in the leaves. The reduced accumulation is caused by lower sulfate uptake and translocation to the shoots; however, the flux through the pathway is not affected. In addition, we identified a single nucleotide polymorphism in the OASC gene among A. thaliana accessions that is linked to variation in sulfate content. Both genetic and transgenic complementation confirmed that the exchange of arginine at position 81 for lysine in numerous accessions resulted in a less active OASC and a lower sulfate content in the leaves. The mitochondrial isoform of OAS-TL is, thus, after the ATPS1 isoform of sulfurylase and the APR2 form of APS reductase 2, the next metabolic enzyme with a role in regulation of sulfate content in Arabidopsis.

Sulfur dioxide mitigates oxidative damage by modulating hydrogen peroxide homeostasis in postharvest table grapes

The hydrogen peroxide (H2O2) burst accelerates the decay and senescence of table grapes. Sulfur dioxide (SO2), a commercial preservative, is known to maintain the postharvest quality of table grapes. However, the mechanism by which SO2 improves postharvest quality is still unclear. In this study, the effects of SO2 on sulfite reduction, the ascorbate-glutathione (AsA-GSH) cycle and the H2O2 receptor during storage were evaluated. The results showed that SO2 not only inhibited decay but also reduced the accumulation of H2O2 and malondialdehyde (MDA). Compared to the control, SO2 treatment increased the activities of sulfite reductase (SiR), serine acetyltransferase (SAT) and O-acetylserine (thiol)-lyase (OASTL), the content of cysteine (Cys) and the expression levels of VvSiR, VvSAT1, VvSAT2 and VvOASTL. Enhancement of critical enzyme activity, gene expression and antioxidant content in the AsA-GSH cycle was observed in SO2-treated grapes. Furthermore, VvGS expression was also determined to be involved in GSH synthesis. SO2 repressed the transcription level of VvHPCA1–4, and VvHPCA3 could be used as a candidate gene to evaluate the degree of oxidative damage in table grapes. These findings suggested that SO2 maintained H2O2 homeostasis to alleviate postharvest oxidative damage by activating sulfite reduction and the AsA-GSH cycle along with inhibiting the expression of VvHPCAs.

Impact of heavy metals in the microalga Chlorella sorokiniana and assessment of its potential use in cadmium bioremediation.

The chlorophyte microalga Chlorella sorokiniana was tested for the bioremediation of heavy metals pollution. It was cultured with different concentrations of Cu2+, Cd2+, As (III) and As (V), showing a significant inhibition on its growth at concentrations of 500 µM Cu2+, 250 µM Cd2+, 750 µM AsO33- and 5 mM AsO43- or higher. Moreover, the consumption of ammonium was also studied, showing significant differences for concentrations higher than 1 mM of Cu2+ and As (III), and 5 mM of As (V). The determination of intracellular heavy metals concentration revealed that Chlorella sorokiniana is an outstanding Cd accumulator organism, able to accumulate 11,232 mg kg-1 of Cd, and removing 65% of initial concentration of this heavy metal. Finally, antioxidant enzymes, such as catalase (CAT) and ascorbate peroxidase (APX), and enzymes involved in the production of glutamate and cysteine, such as glutamine syntethase (GS), glutamate dehydrogenase (GDH), O-acetylserine (thiol) lyase (OASTL) and NAD-isocitrate dehydrogenase (NAD-IDH) were studied both at gene expression and enzymatic activity levels. These enzymes exhibited different grades of upregulation, especially in response to Cd and As stress. However, GS expression was downregulated when Chlorella sorokiniana was cultured in the presence of these heavy metals.

Bioinformatics analysis of enzymes involved in cysteine biosynthesis: first evidence for the formation of cysteine synthase complex in cyanobacteria.

The online version contains supplementary material available at 10.1007/s13205-021-02899-1.

Chickpea glutaredoxin (CaGrx) gene mitigates drought and salinity stress by modulating the physiological performance and antioxidant defense mechanisms.

The online version contains supplementary material available at 10.1007/s12298-021-00999-z.

Response of Olive Shoots to Salinity Stress Suggests the Involvement of Sulfur Metabolism.

Global warming has two dangerous global consequences for agriculture: drought, due to water scarcity, and salinization, due to the prolonged use of water containing high concentrations of salts. Since the global climate is projected to continue to change over this century and beyond, choosing salt-tolerant plants could represent a potential paramount last resort for exploiting the secondary saline soils. Olive is considered moderately resistant to soil salinity as compared to other fruit trees, and in the present study, we investigated the influence of NaCl solutions (ranging from 0 to 200 mM) in a salt-tolerant (cv Canino) and two of its transgenic lines (Canino AT17-1 and Canino AT17-2), overexpressing tobacco osmotin gene, and in a salt-sensitive (Sirole) olive cultivar. After four weeks, most of the shoots of both Canino and Sirole plants showed stunted growth and ultimate leaf drop by exposure to salt-enriched media, contrary to transgenic lines, that did not show injuries and exhibited a normal growth rate. Malondialdehyde (MDA) content was also measured as an indicator of the lipid peroxidation level. To evaluate the role of the S assimilatory pathway in alleviating the adverse effects of salt stress, thiols levels as well as extractable activities of ATP sulfurylase (ATPS) and O-acetyl serine(thiol)lyase (OASTL), the first and the last enzyme of the S assimilation pathway, respectively, have been estimated. The results have clearly depicted that both transgenic lines overexpressing osmotin gene coped with increasing levels of NaCl by the induction of S metabolism, and particularly increase in OASTL activity closely paralleled changes of NaCl concentration. Linear correlation between salt stress and OASTL activity provides evidence that the S assimilation pathway plays a key role in adaptive response of olive plants under salt stress conditions.

Molecular cloning of putative chloroplastic cysteine synthase in Leucaena leucocephala.

Cysteine biosynthesis is directed by the successive commitments of serine acetyltransferase, and O-acetylserine (thiol) lyase (OASTL) compounds, which subsequently frame the decameric cysteine synthase complex. The isoforms of OASTL are found in three compartments of the cell: the cytosol, plastid, and mitochondria. In this investigation, we first isolated putative chloroplastic OASTL (Ch-OASTL) from Leucaena leucocephala, and the Ch-OASTL was then expressed in BL21-competent Escherichia coli. The putative Ch-OASTL cDNA clone had 1,543 base pairs with 391 amino acids in its open reading frame and a molecular weight of 41.54kDa. The purified protein product exhibited cysteine synthesis ability, but not mimosine synthesis activity. However, they both make the common α-aminoacrylate intermediate in their first half reaction scheme with the conventional substrate O-acetyl serine (OAS). Hence, we considered putative Ch-OASTL a cysteine-specific enzyme. Kinetic studies demonstrated that the optimum pH for cysteine synthesis was 7.0, and the optimum temperature was 40°C. In the cysteine synthesis assay, the Km and kcat values were 838 ± 26µM and 72.83s-1 for OAS, respectively, and 60 ± 2µM and 2.43s-1 for Na2S, respectively. We can infer that putative Ch-OASTL regulatory role is considered a sensor for sulfur constraint conditions, and it acts as a forerunner of various metabolic compound molecules.

Regulation of Hydrogen Sulfide Metabolism by Nitric Oxide Inhibitors and the Quality of Peaches during Cold Storage.

Both nitric oxide (NO) and hydrogen sulfide (H2S) have been shown to have positive effects on the maintenance of fruit quality during storage; however, the mechanisms by which NO regulates the endogenous H2S metabolism remain unknown. In this experiment, peaches were immersed in solutions of NO, potassium 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO, as an NO scavenger), N-nitro-l-arginine methyl ester (l-NAME, as an inhibitor of nitric oxide synthase (NOS)-like activity), and sodium tungstate (as an inhibitor of nitrate reductase), and the resulting changes in the H2S metabolism of peaches were studied. The results showed that exogenous NO reduced the contents of endogenous H2S, Cys, and sulfite; decreased the activities of l-/d-cysteine desulfhydrase (l-/d-CD), O-acetylserine (thiol)lyase (OAS-TL), and sulfite reductase (SiR); and increased the activity of β-cyanoalanine synthase (β-CAS). Both c-PTIO and sodium tungstate had similar roles in increasing the H2S content by sustaining the activities of l-/d-CDs, OAS-TL, and SiR. l-NAME increased the H2S content, mainly by maintaining the d-CD activity. The results suggest that NO, c-PTIO, l-NAME, and sodium tungstate differently regulate the H2S metabolism of peaches during storage.

Crop adaptation to air pollution ii. tolerance to so2 stress is regulated by oxidative and antioxidative characteristics and sulphur assimilation

Sulphur dioxide (SO2) and particulate matter (PM) are one of the major air pollutants emerging out of the industrial development and human activities. Plants exhibit differential sensitivity to SO2 pollution and its effects on plant growth can be both direct and/or indirect. We have earlier reported that a high SO2 stress contributes toward the S-nutrition of crops. The SO2 enriched environment significantly improved the activity of serine transacetylase (SAT) in all the experimental crops, however, the activity of O-acetylserine (thiol) lyase (OAS-TL) was enhanced chiefly in wheat but not in chickpea and barley. Further, the relative tolerance of crops to the particulate and gaseous pollutants was related to a lower level of superoxide and H2O2 radicals and lipid peroxidation and a higher level of antioxidants such as ascorbic acid and peroxidase activity. Relative tolerance of crops to the particulate and gaseous pollutants was related to a lower oxidative stress and a higher anti-oxidative defence that elevated SO2 contributes to S-nutrition of crops however, the threshold value of phyto-toxicity need to be determined across the crops.

Cloning and functional analysis of four O-Acetylserine (thiol) lyase family genes from foxtail millet

Cysteine is the first organic molecule generated during the assimilation of sulfate. As such, cysteine and its derivatives are always essential signal molecules and thus have important roles in the regulation of many plant processes. O-acetylserine (thiol) lyase (OASTL) catalyzes the last step of the biosynthesis of cysteine. At present, detailed and comprehensive work about these enzymes has only been reported from the plant Arabidopsis thaliana, though sporadic studies on OASTL have been conducted on other dicots, such as spinach and soybean. However, few reports on the functions of OASTLs in monocots have been found in the literature. Here in this study, we obtained four SiOASTL genes (SiOASTL7, SiOASTL8, SiOASTL9 and SiOASTL10) from foxtail millet and analyzed their potential functions. Phylogenetically, the four SiOASTL genes did not belong to any published subfamily of the OASTL genes; instead they constituted a new subfamily specific to the OASTL genes from monocots. In sequencing, we found that with the exception of the pseudogene SiOASTL8, proteins encoded by the other three genes exhibited high similarity with OASTL proteins from Arabidopsis, though the critical PLP-binding sites of both SiOASTL7 and SiOASTL10 were missing. The enzymatic activity assays demonstrated that SiOASTL9 has the ability to catalyze the biosynthesis of both cysteine and S-sulfocysteine, while SiOASTL7 and SiOASTL10 did not possess any previously reported catalyzing abilities. In addition, the gene expression pattern analysis showed that all four genes were widely expressed in various tissues of foxtail millet, and all had a preference in the leaves. Under abiotic stresses, the expression of these genes could be induced by salt and drought stress. Our finding that cadmium could only up-regulate the transcription of SlOASTL8 and SlOASTL9, further indicates the diversified responses of SiOASTLs to abiotic stresses.

Thiol redox-regulation for efficient adjustment of sulfur metabolism in acclimation to abiotic stress.

Sulfur assimilation and sulfur metabolism are tightly controlled at the transcriptional, post-transcriptional, and post-translational levels in order to meet the demand for reduced sulfur in growth and metabolism. These regulatory mechanisms coordinate the cellular sulfhydryl supply with carbon and nitrogen assimilation in particular. Redox homeostasis is an important cellular parameter intimately connected to sulfur by means of multiple thiol modifications. Post-translational thiol modifications such as disulfide formation, sulfenylation, S-nitrosylation, persulfidation, and S-glutathionylation allow for versatile switching and adjustment of protein functions. This review focuses on redox-regulation of enzymes involved in the sulfur assimilation pathway, namely adenosine 5´-phosphosulfate reductase (APR), adenosine 5´-phosphosulfate kinase (APSK), and γ-glutamylcysteine ligase (GCL). The activity of these enzymes is adjusted at the transcriptional and post-translational level depending on physiological requirements and the state of the redox and reactive oxygen species network, which are tightly linked to abiotic stress conditions. Hormone-dependent fine-tuning contributes to regulation of sulfur assimilation. Thus, the link between oxylipin signalling and sulfur assimilation has been substantiated by identification of the so-called COPS module in the chloroplast with its components cyclophilin 20-3, O-acetylserine thiol lyase, 2-cysteine peroxiredoxin, and serine acetyl transferase. We now have a detailed understanding of how regulation enables the fine-tuning of sulfur assimilation under both normal and abiotic stress conditions.

GRA78 encoding a putative S-sulfocysteine synthase is involved in chloroplast development at the early seedling stage of rice

Cysteine functions not only as an amino acid in proteins but also as a precursor for a large number of essential biomolecules. Cysteine is synthesized via the incorporation of sulfide to O-acetylserine under the catalysis of O-acetylserine(thiol)lyase (OASTL). In dicotyledonous Arabidopsis, nine OASTL genes have been reported. However, in their null mutants, only the mutant of CS26 encoding S-sulfocysteine synthase showed the visible phenotypic changes, displaying significantly small plants and pale-green leaves under long-day condition but not short-day condition. Up to now, no OASTL gene or mutant has been identified in monocotyledon. In this study, we isolated a green-revertible albino mutant gra78 in rice (Oryza sativa). Its albino phenotype at the early seedling stage was sensitive to temperature but independent of photoperiod. Map-based cloning revealed that candidate gene LOC_Os01g59920 of GRA78 encodes a putative S-sulfocysteine synthase showing significant similarity with Arabidopsis CS26. Complementation experiment confirmed that mutation in LOC_Os01g59920 accounted for the mutant phenotype of gra78. GRA78 is constitutively expressed in all tissues and its encoded protein is targeted to the chloroplast. In addition, qRT-PCR suggested that expression levels of four OASTL homolog genes and five photosynthetic genes were remarkably down-regulated.

Hydrogen Sulfide-Mediated Activation of O-Acetylserine (Thiol) Lyase and l/d-Cysteine Desulfhydrase Enhance Dehydration Tolerance in Eruca sativa Mill.

Hydrogen sulfide (H2S) has emerged as an important signaling molecule and plays a significant role during different environmental stresses in plants. The present work was carried out to explore the potential role of H2S in reversal of dehydration stress-inhibited O-acetylserine (thiol) lyase (OAS-TL), l-cysteine desulfhydrase (LCD), and d-cysteine desulfhydrase (DCD) response in arugula (Eruca sativa Mill.) plants. Dehydration-stressed plants exhibited reduced water status and increased levels of hydrogen peroxide (H2O2) and superoxide (O2•−) content that increased membrane permeability and lipid peroxidation, and caused a reduction in chlorophyll content. However, H2S donor sodium hydrosulfide (NaHS), at the rate of 2 mM, substantially reduced oxidative stress (lower H2O2 and O2•−) by upregulating activities of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase) and increasing accumulation of osmolytes viz. proline and glycine betaine (GB). All these, together, resulted in reduced membrane permeability, lipid peroxidation, water loss, and improved hydration level of plants. The beneficial role of H2S in the tolerance of plants to dehydration stress was traced with H2S-mediated activation of carbonic anhydrase activity and enzyme involved in the biosynthesis of cysteine (Cys), such as OAS-TL. H2S-treated plants showed maximum Cys content. The exogenous application of H2S also induced the activity of LCD and DCD enzymes that assisted the plants to synthesize more H2S from accumulated Cys. Therefore, an adequate concentration of H2S was maintained, that improved the efficiency of plants to mitigate dehydration stress-induced alterations. The central role of H2S in the reversal of dehydration stress-induced damage was evident with the use of the H2S scavenger, hypotaurine.