APS Reductase Research Articles

Potassium and jasmonic acid —Induced nitrogen and sulfur metabolisms improve resilience against arsenate toxicity in tomato seedlings

Despite the well-known individual roles of potassium (K+) and jasmonic acid (JA) in plant responses to various stresses, their combined effects on improving tolerance to metalloid arsenate (AsV) toxicity and their influence on physiological and biochemical mechanisms remain largely unexplored. Therefore, the present study was conducted to explore the concomitant role of K+ and JA in nitrogen (N) and sulfur (S) metabolisms and regulation of antioxidant system in tomato seedlings under AsV toxicity conditions. The obtained data reveal that K+ and/or JA boosted the tolerance of tomato (Solanum lycopersicum L.) seedlings to AsV toxicity. However, the effect of a combined dose of K+ and JA was found to be more efficient as compared to the alone application. Seedlings subjected to AsV exhibited suppressed morphological attributes (shoot and root length, shoot and root fresh weight, and shoot and root dry weight), and physio-biochemical characteristics. Also, AsV-treated tomato seedlings showed enhanced reactive oxygen species (ROS) and increased the activity of ROS-producing enzymes (O2•— -generating NADPH oxidase activity and H2O2-generating glycolate oxidase] and also a chlorophyll (Chl) degrading enzyme [chlorophyllase (Chlase) activity]. However, the exogenous supply of K+ and JA synergistically inhibited Chl degradation and overproduction of ROS by suppressing their responsible enzymes activity. Moreover, it induced gas exchange parameters and activity of enzymes responsible for the photosynthesis process (carbonic anhydrase and Rubisco), Chl biosynthesis (δ-aminolevulinic acid dehydratase), and suppressed Chlase activity, and overproduction of ROS and their producing enzymes activity. Most interestingly, the combined application of K+ and JA substantially increased N and S content and their assimilation by upregulating the activity of enzymes involved in these key pathways (nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase, ATP sulfurylase, adenosine 5-phosphosulfate reductase, O-acetylserine (thiol) lyase and sulfide reductase). The obtained results strongly reveal that the combined application of K+ and JA improved tolerance of tomato seedlings to AsV toxicity by enhancing N and S assimilation pathways and antioxidant system.

A cotton APS REDUCTASE represses root hair elongation via sulfur assimilation and hydrogen peroxide‐mediated cell wall damage

AbstractSulfur is a critical element for plant growth and development, and it is assimilated through a pathway called the sulfur assimilation pathway. APS REDUCTASE (APR) is a key rate‐limiting enzyme in this pathway, and it plays a vital role in regulating sulfur flow and plant development. However, the exact role and mechanism of APR in cotton fiber and root hair development are still not fully understood. This study used in vitro cultured ovules grown on a sulfur‐deficient medium to explore the role of sulfur in fiber development. Genetic approaches such as virus‐induced gene silencing (VIGS) was used to determine the role of target genes. In vitro assays revealed the role of sulfur in fiber development and identified GhAPR2 as a key candidate gene. The GhAPR2 negatively regulates fiber cell initiation and Arabidopsis root hair elongation. Further studies demonstrated that overexpression of GhAPR2 increased the contents of sulfite and GSH to enhance sulfur assimilation in Arabidopsis, leading to abnormal accumulation of ROS in root hair. This down‐regulates the transcription of proline‐rich cell wall protein‐coding genes such as AtEXTs and AtPRPs, reducing the content of hydroxyproline in the cell wall, disrupting the development of the root hair cell wall, and inhibiting root hair elongation. This study elucidated the functional pathway of APR2 in the regulation of root hair development. APR2 first affects sulfur assimilation, which affects the content of reactive oxygen species (ROS), and finally affects the content of hydroxyproline‐rich glycoprotein, which in turn affects the development of root hair. This model also contributes to the understanding of the role of sulfur in fiber development, as well as cell differentiation and development.

Biodegradation of sulfate and elimination of heavy metals by immobilized-microbial bioaugmentation coupled with anaerobic membrane bioreactor

Sulfate-rich metal-laden wastewater (SRMLW) shows a significant threat on the ecological environment and human health. Sulfate-reducing bacteria (SRB) can remove sulfate from wastewater in anaerobic membrane bioreactor (AnMBR). This study constructed recombinant genetic engineered SRB (rSRB) and investigated the sulfate and heavy metals removal efficiency for treating SRMLW by immobilized-microbial bioaugmentation using rSRB (IMrSRB) coupled with AnMBR (Bio-AnMBR). The results show that Bio-AnMBR removed sulfate (99.4%) from SRMLW by upregulating key genes (Sat, DsrAB, AprAB, QmoABC, DsrC and DsrMKJOP) and enzymes (APS reductase, Dissimilatory sulfite reductase, Trithionate reductase and Thiosulfate reductase) in dissimilatory sulfite and sulfate respiration processes. Bio-AnMBR achieved synergistic biodegradation of sulfate and nitrogen by stimulating the activity of these bacteria related to the conversion of nitrogen and sulfate. Microbial analysis indicated that the relatively high abundance of rSRB (79.2%) and functional genes (70.1%) upregulated the activities of key enzymes associated with sulfate metabolism system, thus increasing the production of S (66.4 mg/L) and H2S (23.8 mg/L). Bio-AnMBR significantly improved the tolerance to heavy metals by increasing concentrations of CO32–, S2-, and HS-, which was configured to reduce heavy metals (95.6%) via precipitation of metal sulfide (81.3 mg/L), metal carbonates (30.7 mg/L) and metal hydroxides (18.7 mg/L). Bio-AnMBR increased system stability, shorted hydraulic retention time (HRT), decreased the production of soluble microbial products (SMP), extracellular polymeric substances (EPS) and waste-sludge production (WSP), as well as improved effluent quality. In conclusion, data in this study indicate that application of Bio-AnMBR is a feasible strategy for the treatment of SRMLW.

Distribution and Activity of Sulfur-Metabolizing Bacteria along the Temperature Gradient in Phototrophic Mats of the Chilean Hot Spring Porcelana.

In terrestrial hot springs, some members of the microbial mat community utilize sulfur chemical species for reduction and oxidization metabolism. In this study, the diversity and activity of sulfur-metabolizing bacteria were evaluated along a temperature gradient (48-69 °C) in non-acidic phototrophic mats of the Porcelana hot spring (Northern Patagonia, Chile) using complementary meta-omic methodologies and specific amplification of the aprA (APS reductase) and soxB (thiosulfohydrolase) genes. Overall, the key players in sulfur metabolism varied mostly in abundance along the temperature gradient, which is relevant for evaluating the possible implications of microorganisms associated with sulfur cycling under the current global climate change scenario. Our results strongly suggest that sulfate reduction occurs throughout the whole temperature gradient, being supported by different taxa depending on temperature. Assimilative sulfate reduction is the most relevant pathway in terms of taxonomic abundance and activity, whereas the sulfur-oxidizing system (Sox) is likely to be more diverse at low rather than at high temperatures. Members of the phylum Chloroflexota showed higher sulfur cycle-related transcriptional activity at 66 °C, with a potential contribution to sulfate reduction and oxidation to thiosulfate. In contrast, at the lowest temperature (48 °C), Burkholderiales and Acetobacterales (both Pseudomonadota, also known as Proteobacteria) showed a higher contribution to dissimilative sulfate reduction/oxidation as well as to thiosulfate metabolism. Cyanobacteriota and Planctomycetota were especially active in assimilatory sulfate reduction. Analysis of the aprA and soxB genes pointed to members of the order Burkholderiales (Gammaproteobacteria) as the most dominant and active along the temperature gradient for these genes. Changes in the diversity and activity of different sulfur-metabolizing bacteria in photoautotrophic microbial mats along a temperature gradient revealed their important role in hot spring environments, especially the main primary producers (Chloroflexota/Cyanobacteriota) and diazotrophs (Cyanobacteriota), showing that carbon, nitrogen, and sulfur cycles are highly linked in these extreme systems.

Reduced sulfide and methane in rising main sewer via calcium peroxide dosing: Insights from microbial physiological characteristics, metabolisms and community traits

Although the biocidal effect of calcium peroxide (CaO2) has attracted increasing attention in wastewater and sludge management, its potential in the reduction of sulfide and methane from sewer is not tapped. This study aims to fill this gap through the long-term operated sewer reactors. Results showed one-time dose of 0.2% (w/v) CaO2 with 12-h exposure decreased the average sulfide and methane production by 80% during one week. The electron paramagnetic resonance and free radical quenching tests indicated free radicals from CaO2 decomposing posed a major contribution on sewer biofilms (•OH>•O2−>alkali). Mechanistic analysis revealed extracellular polymeric matrix breakdown (e.g., protein secondary structure) and cell membrane damage were caused by the increased lipid peroxidation of cells and exacerbated intracellular reactive oxygen species under CaO2 stress. Moreover, the intracellular metabolic pathways, such as electrons provision and transfer, as well as pivotal enzymatic activities (e.g., APS reductase, sulfite reductase and coenzymes F420) were significantly impaired. RT-qPCR analysis unveiled the absolute abundances of dsrA and mcrA were decreased by 7.53–40.37% and 67.00–74.85%, respectively. Although this study broadens the application scope of CaO2 and provides in-depth understanding of advanced oxidation-based technology in sewer management, the pipe scale risk due to the release of calcium ions warrants further investigation.

Potential Inhibitor of Adenylyl Sulfate Reductase Isolated from Desulfovibrio desulfuricans with PU/PU-Ag to Control Pitting Corrosion of Oil Tanks and Pipelines

Background and Aim: This study aims to alleviate the microbiologically affected corrosion that occurred by sulfate-reducing bacteria (SRB) through synthesizing a bio-based polyurethane polymer and its nanocomposite coating, silver nanoparticles (PU-Ag). Moreover, it will also evaluate the effect of PU alone and PU-Ag as inhibitors for adenylyl sulfate reductase (APS), the main enzyme for sulfate reduction. Methods: In this study, the PU was prepared from vegetable soybean oil, and the silver nanoparticles (Ag-NPs) with a concentration of 1% were coated to the PU forming a nanocomposite. The PU and the PU-Ag were characterized and evaluated as inhibitors of the APS reductase enzyme. Results: The results obtained from FTIR, UV, DLS, TEM, and XRD confirmed the preparation structure of the PU and PU-Ag. Furthermore, the PU/PU-Ag competitively inhibited the APS reductase with an inhibition constant equal to 35.7 and 11 mg, respectively. These indicated the exerted inhibitory effect of PU/PU-Ag upon the activity of the APS reductase enzyme. Conclusion: The APS reductase enzyme produced by SRB, which is recorded as a big problem in the oil and gas industry such as pitting corrosion of tanks and pipelines, could be inhibited by PU and PU-Ag.

De novo Transcriptome Assembly and Comparative Analysis Highlight the Primary Mechanism Regulating the Response to Selenium Stimuli in Oats (Avena sativa L.).

Selenium is an essential microelement for humans and animals. The specific processing technique of oats can maximize the preservation of its nutrients. In this study, to understand the genetic response of oats in a high-selenium environment, oats were treated with sodium selenate for 24 h, and transcriptome analysis was performed. A total of 211,485,930 clean reads composing 31.30 Gb of clean data were retained for four samples. After assembly, 186,035 unigenes with an average length of 727 bp were generated, and the N50 length was 1,149 bp. Compared with that in the control group, the expression of 7,226 unigenes in the treatment group was upregulated, and 2,618 unigenes were downregulated. Based on the sulfur assimilation pathway and selenocompound metabolic pathway, a total of 27 unigenes related to selenate metabolism were identified. Among them, the expression of both key genes APS (ATP sulfurylase) and APR (adenosine 5′-phosphosulfate reductase) was upregulated more than 1,000-fold under selenate treatment, while that of CBL (cystathionine-β-synthase) was upregulated 3.12-fold. Based on the transcriptome analysis, we suspect that the high-affinity sulfur transporter Sultr1;2 plays a key role in selenate uptake in oats. A preliminary regulatory mechanism explains the oat response to selenate treatment was ultimately proposed based on the transcriptome analysis and previous research.

Crystal Structure of the [4Fe-4S] Cluster-Containing Adenosine-5'-phosphosulfate Reductase from Mycobacterium tuberculosis.

Tuberculosis (TB) is the deadliest infectious disease in the world. In Mycobacterium tuberculosis, the first committed step in sulfate assimilation is the reductive cleavage of adenosine-5′-phosphosulfate (APS) to form adenosine-5′-phosphate (AMP) and sulfite by the enzyme APS reductase (APSR). The vital role of APSR in the production of essential reduced-sulfur-containing metabolites and the absence of a homologue enzyme in humans makes APSR a potential target for therapeutic interventions. Here, we present the crystal structure of the [4Fe–4S] cluster-containing APSR from M. tuberculosis (MtbAPSR) and compare it to previously determined structures of sulfonucleotide reductases. We further present MtbAPSR structures with substrate APS and product AMP bound in the active site. Our structures at a 3.1 Å resolution show high structural similarity to other sulfonucleotide reductases and reveal that APS and AMP have similar binding modes. These studies provide structural data for structure-based drug design aimed to combat TB.

3D-QSAR study of adenosine 5’-phosphosulfate (APS) analouges as ligands for APS reductase

Metabolism of sulfur (sulfur assimilation pathway, SAP) is one of the key pathways for the pathogenesis and survival of persistant bacteria, such as Mycobacterium tuberculosis (Mtb), in the latent period. Adenosine 5'-phosphosulfate reductase (APSR) is an important enzyme involved in the SAP, absent from the human body, so it might represent a valid target for development of new antituberculosis drugs. This work aimed to develop 3D-QSAR model based on the crystal structure of APSR from Pseudomonas aeruginosa, which shows high degree of homology with APSR from Mtb, in complex with its substrate, adenosine 5'-phosphosulfate (APS). 3D-QSAR model was built from a set of 16 nucleotide analogues of APS using alignment-independent descriptors derived from molecular interaction fields (MIF). The model improves the understanding of the key characteristics of molecules necessary for the interaction with target, and enables the rational design of novel small molecule inhibitors of Mtb APSR.

Reaction mechanism catalyzed by the dissimilatory adenosine 5′-phosphosulfate reductase. Adenosine 5′-monophosphate inhibitor and key role of arginine 317 in switching the course of catalysis

The present research is a continuation of our work on dissimilatory reduction pathway of sulfate - involved in biogeochemical sulfur turnover. Adenosine 5′-phosphosulfate reductase (APSR) is the second enzyme in the dissimilatory pathway of the sulfate to sulfide reduction. It reversibly catalyzes formation of the sulfite anion (HSO3−) from adenosine 5′-phosphosulfate (APS) - the activated form of sulfate provided by ATP sulfurylase (ATPS). Two electrons required for this redox reaction derive from reduced FAD cofactor, which is suggested to be involved directly in the catalysis by formation of FADH-SO3− intermediate. The present work covers quantum-mechanical (QM) studies on APSR reaction performed for eight models of APSR active site. The cluster models were constructed based on two crystal structures (PDB codes: 2FJA and 2FJB), differing in conformation of Arg317 active site residue. The described results indicated the most feasible mechanism of APSR forward reaction, including formation of FADHN-SO3− adduct (with proton on N5 atom of isoalloxazine), tautomerization of FADHN-SO3− to FADHO-SO3− (with proton on CO moiety of isoalloxazine), and its reductive cleavage to oxidized FAD and sulfite anion. The reverse reaction proceeds in the backward direction. It is suggested that it requires two AMP molecules, one acting as a substrate and another as an inhibitor of forward reaction, which forces change of Arg317 conformation from “arginine in” (2FJA) to “arginine out” (2FJB). Important role of Arg317 in switching the course of the APSR catalytic reaction is revealed by changing the direction of thermodynamic driving force. The presented research also shows the importance of the protonation pattern of the reduced FAD cofactor and protein residues within the active site.

Development and Demonstration of Membrane to Control Potential Pathogen (Legionella sp.) Associated With Cooling Towers

The presence of pathogens in industrial cooling towers has been identified as one of the causes of legionellosis, but the tangible hazard has been unrecognized. The major function of a cooling tower in the industries is to transfer heat into the environment through evaporation which leads to an unusual increment in the conductivity of water. Conductivity indicates dissolved solids of water which further facilitate the occurrence of corrosion within the cooling tower systems that may hamper the overall functioning of the cooling tower. The lab-scale study was conducted to developed and demonstrated an efficient membrane to control potential pathogen (Legionella sp) associated with cooling tower water. In the present study, cooling tower water from various locations was collected and examined for physicochemical parameters (pH, conductivity, salinity, sulphate, & APS reductase) and microbial load. The phylogenetic analysis of the potential sample (with a maximum load of bacterial counts, P1) confirms the presence of Legionella sp., Pseudomonas sp. and Flavobacterium sp. In the present investigation, a unique membrane was developed to regulate the growth of pathogenic contaminants along with sulphate, alkalinity in the cooling tower water. The membrane was capable of reducing conductivity from 6.03 to 4.5 mS/cm and total microbial load reduced by 40% by passing through the liquid. The developed membrane was characterized through FTIR that indicated the presence of carbonyl, C-CH3 and C-O-C group whereas, thermal properties of the membrane was determined through TGA. Thus the developed membrane may be used for environmental monitoring and maintenance, including anti-scale treatment, and physical, chemical, and microbiological control, ensure the good performance of reducing the Legionella sp risk for public health in cooling tower stations.

Regulation of Sulfate Uptake and Assimilation in Barley (Hordeum vulgare) as Affected by Rhizospheric and Atmospheric Sulfur Nutrition.

To study the regulation of sulfate metabolism in barley (Hordeum vulgare), seedlings were exposed to atmospheric hydrogen sulfide (H2S) in the presence and absence of a sulfate supply. Sulfate deprivation reduced shoot and root biomass production by 60% and 70%, respectively, and it affected the plant’s mineral nutrient composition. It resulted in a 5.7- and 2.9-fold increased shoot and root molybdenum content, respectively, and a decreased content of several other mineral nutrients. Particularly, it decreased shoot and root total sulfur contents by 60% and 70%, respectively. These decreases could be ascribed to decreased sulfate contents. Sulfate deficiency was additionally characterized by significantly lowered cysteine, glutathione and soluble protein levels, enhanced dry matter, nitrate and free amino acid contents, an increased APS reductase (APR) activity and an increased expression and activity of the root sulfate uptake transporters. When sulfate-deprived barley was exposed to 0.6 µL L−1 atmospheric H2S, the decrease in biomass production and the development of other sulfur deficiency symptoms were alleviated. Clearly, barley could use H2S, absorbed by the foliage, as a sulfur source for growth. H2S fumigation of both sulfate-deprived and sulfate-sufficient plants downregulated APR activity as well as the expression and activity of the sulfate uptake transporters. Evidently, barley switched from rhizospheric sulfate to atmospheric H2S as sulfur source. Though this indicates that sulfate utilization in barley is controlled by signals originating in the shoot, the signal transduction pathway involved in the shoot-to-root regulation must be further elucidated.

Exogenous nitric oxide requires endogenous hydrogen sulfide to induce the resilience through sulfur assimilation in tomato seedlings under hexavalent chromium toxicity

Nitric oxide (NO) and hydrogen sulfide (H2S), versatile signaling molecules, play multiple roles in plant growth, physiological and biochemical processes under heavy metal stress. However, the mechanisms through which NO in association with endogenous H2S mediated hexavalent chromium Cr(VI) toxicity mitigation are still not fully understood. Therefore, we investigated the role of NO and H2S in sulfur (S)-assimilation and the effect of NO on endogenous H2S, and cysteine (Cys) biosynthesis and maintenance of cellular glutathione (GSH) pool in tomato seedlings under Cr(VI) stress. Cr(VI) toxicity caused an increase in reactive oxygen species (ROS; O2•– and H2O2) formation and activity of chlorophyll (Chl) degrading enzyme [Chlorophyllase (Chlase)] and decrease in seedlings growth attributes, Chl a and b content, and activity of Chl synthesizing enzyme [δ-aminolevulinic acid dehydratase (δ-ALAD)], gas exchange parameters, S-assimilation, and Cys and H2S metabolism. An increase in the content of glycinebetaine (GB), total soluble carbohydrates (TSCs) and total phenols (TPls), and decrease in DNA damage and ROS in NO treated seedlings conferred Cr(VI) toxicity tolerance. Under Cr(VI) toxicity conditions, the inclusion of H2S scavenger hypotaurine (HT) in growth medium containing NO validated the role of endogenous H2S in S-assimilation, H2S and Cys and GSH metabolism by withdrawing activity of enzymes involved in S-assimilation [adenosine 5-phosphosulfatereductase (APS-R), ATP-sulfurylase (ATP-S)], in the biosynthesis of H2S [L-cysteine desulfhydrase (L-CD) and D-cysteine desulfhydrase (D-CD)], Cys [O-acetylserin (thiol) lyase (OAST-L)], and GSH [glutamylcysteine synthetase (γ-GCS) and glutathione synthetase (GS)], and in antioxidant system. On the other hand, application of cPTIO [2-(4–32 carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide], a NO scavenger and HT diminished the effect of NO on internal H2S levels, Cys and glutathione homeostasis, and S-assimilation, which resulted in poor immunity against oxidative stress induced by Cr(VI) toxicity. The obtained results postulate that NO-induced internal H2S conferred tolerance of tomato seedlings to Cr(VI) toxicity and maintained better photosynthesis process and plant growth.

Adenosine-5'-Phosphosulfate- and Sulfite Reductases Activities of Sulfate-Reducing Bacteria from Various Environments.

A comparative study of the kinetic characteristics (specific activity, initial and maximum rate, and affinity for substrates) of key enzymes of assimilatory sulfate reduction (APS reductase and dissimilatory sulfite reductase) in cell-free extracts of sulphate-reducing bacteria (SRB) from various biotopes was performed. The material for the study represented different strains of SRB from various ecotopes. Microbiological (isolation and cultivation), biochemical (free cell extract preparation) and chemical (enzyme activity determination) methods served in defining kinetic characteristics of SRB enzymes. The determined affinity data for substrates (i.e., sulfite) were 10 times higher for SRB strains isolated from environmental (soil) ecotopes than for strains from the human intestine. The maximum rate of APS reductase reached 0.282–0.862 µmol/min×mg−1 of protein that is only 10 to 28% higher than similar initial values. The maximum rate of sulfite reductase for corrosive relevant collection strains and SRB strains isolated from heating systems were increased by 3 to 10 times. A completely different picture was found for the intestinal SRB Vmax in the strains Desulfovibrio piger Vib-7 (0.67 µmol/min × mg−1 protein) and Desulfomicrobium orale Rod-9 (0.45 µmol/min × mg−1 protein). The determinant in the cluster distribution of SRB strains is the activity of the terminal enzyme of dissimilatory sulfate reduction—sulfite reductase, but not APS reductase. The data obtained from the activity of sulfate reduction enzymes indicated the adaptive plasticity of SRB strains that is manifested in the change in enzymatic activity.

Evidence of a significant role of glutathione reductase in the sulfur assimilation pathway.

With the objective of studying the role of glutathione reductase (GR) in the accumulation of cysteine and methionine, we generated transgenic tobacco and Arabidopsis lines overexpressing the cytosolic AtGR1 and the plastidic AtGR2 genes. The transgenic plants had higher contents of cysteine and glutathione. To understand why cysteine levels increased in these plants, we also used gr1 and gr2 mutants. The results showed that the transgenic plants have higher levels of sulfite, cysteine, glutathione and methionine, which are downstream to adenosine 5' phosphosulfate reductase (APR) activity. However, the mutants had lower levels of these metabolites, while the sulfate content increased. A feeding experiment using 34 SO42- also showed that the levels of APR downstream metabolites increased in the transgenic lines and decreased in gr1 compared with their controls. These findings, and the results obtained from the expression levels of several genes related to the sulfur pathway, suggest that GR plays an essential role in the sulfur assimilation pathway by supporting the activity of APR, the key enzyme in this pathway. GR recycles the oxidized form of glutathione (GSSG) back to reduce glutathione (GSH), which serves as an electron donor for APR activity. The phenotypes of the transgenic plants and the mutants are not significantly altered under non-stress and oxidative stress conditions. However, when germinating on sulfur-deficient medium, the transgenic plants grew better, while the mutants were more sensitive than the control plants. The results give substantial evidence of the yet unreported function of GR in the sulfur assimilation pathway.

Differential partitioning of thiols and glucosinolates between shoot and root in Chinese cabbage upon excess zinc exposure

Zinc (Zn) is one of the important elements of plant growth, however, at elevated level it is toxic. Exposure of Chinese cabbage to elevated Zn2+ concentrations (5 and 10 μM ZnCl2) resulted in enhancement of total sulfur and organic sulfur concentration. Transcript level of APS reductase (APR) as a key enzyme in biosynthesis of primary sulfur compounds (cysteine and thiols), was up-regulated in both shoot and root upon exposure to elevated Zn2+, which was accompanied by an increase in the concentration of cysteine in both tissues. In contrast, the concentration of thiols increased only in the root by 5.5 and 15-fold at 5 and 10 μM Zn2+, respectively, which was in accompanied by an upregulation of ATP sulfurylase, an enzyme responsible for activation of sulfate. An elevated content of glucosinolates, mostly indolic glucosinolates, only in the shoot of plants exposed to excess level of Zn2+ coincided with an increase in gene expression of key biosynthetic enzymes and regulators (CYP79B3, CYP83B1, MYB34). Thus distinct acuumulation patterns of sulfur containing compounds in root and shoot of Chinese cabbage may be a strategy for Chinese cabbage to combat with exposure to excess Zn.

Exogenous nitric oxide alleviates sulfur deficiency-induced oxidative damage in tomato seedlings

Despite numerous reports on the role of nitric oxide (NO) in regulating plants growth and mitigating different environmental stresses, its participation in sulfur (S) -metabolism remains largely unknown. Therefore, we studied the role of NO in S acquisition and S-assimilation in tomato seedlings under low S-stress conditions by supplying NO to the leaves of S-sufficient and S-deficient seedlings. S-starved plants exhibited a substantial decreased in plant growth attributes, photosynthetic pigment chlorophyll (Chl) and other photosynthetic parameters, and activity of enzymes involved in Chl biosynthesis (δ-aminolevulinic acid dehydratase), and photosynthetic processes (carbonic anhydrase and RuBisco). Also, S-deficiency enhanced reactive oxygen species (ROS) (superoxide and hydrogen peroxide) and lipid peroxidation (malondialdehyde) levels in tomato seedlings. Contrarily, foliar supplementation of NO to S-deficient seedlings resulted in considerably reduced ROS formation in leaves and roots, which alleviated low S-stress-induced lipid peroxidation. However, exogenous NO enhanced proline accumulation by increasing proline metabolizing enzyme (Δ1-pyrroline-5-carboxylate synthetase) activity and also increased NO, hydrogen sulfide (a gasotransmitter small signaling molecule) and S uptake, and content of S-containing compounds (cysteine and reduced glutathione). Under S-limited conditions, NO improved S utilization efficiency of plants by upregulating the activity of S-assimilating enzymes (ATP sulfurylase, adenosine 5-phosphosulfate reductase, sulfide reductase and O-acetylserine (thiol) lyase). Under S-deprived conditions, improved S-assimilation of seedlings receiving NO resulted in improved redox homeostasis and ascorbate content through increased NO and S uptake. Application of 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxy l-3-oxide (an NO scavenger) invalidated the effect of NO and again caused low S-stress-induced oxidative damage, confirming the beneficial role of NO in seedlings under S-deprived conditions. Thus, exogenous NO enhanced the tolerance of tomato seedlings to limit S-triggered oxidative stress and improved photosynthetic performance and S assimilation.

C-terminal Redox Domain of Arabidopsis APR1 is a Non-Canonical Thioredoxin Domain with Glutaredoxin Function.

Sulfur is an essential nutrient that can be converted into utilizable metabolic forms to produce sulfur-containing metabolites in plant. Adenosine 5′-phosphosulfate (APS) reductase (APR) plays a vital role in catalyzing the reduction of activated sulfate to sulfite, which requires glutathione. Previous studies have shown that the C-terminal domain of APR acts as a glutathione-dependent reductase. The crystal structure of the C-terminal redox domain of Arabidopsis APR1 (AtAPR1) shows a conserved α/β thioredoxin fold, but not a glutaredoxin fold. Further biochemical studies of the redox domain from AtAPR1 provided evidence to support the structural observation. Collectively, our results provide structural and biochemical information to explain how the thioredoxin fold exerts the glutaredoxin function in APR.

Distribution of control in the sulfur assimilation in Arabidopsis thaliana depends on environmental conditions.

Sulfur assimilation is central to the survival of plants and has been studied under different environmental conditions. Multiple studies have been published trying to determine rate-limiting or controlling steps in this pathway. However, the picture remains inconclusive with at least two different enzymes proposed to represent such rate-limiting steps. Here, we used computational modeling to gain an integrative understanding of the distribution of control in the sulfur assimilation pathway of Arabidopsis thaliana. For this purpose, we set up a new ordinary differential equation(ODE)-based, kinetic model of sulfur assimilation encompassing all biochemical reactions directly involved in this process. We fitted the model to published experimental data and produced a model ensemble to deal with parameter uncertainties. The ensemble was validated against additional published experimental data. We used the model ensemble to subsequently analyse the control pattern and robustly identified a set of processes that share the control in this pathway under standard conditions. Interestingly, the pattern of control is dynamic and not static, that is it changes with changing environmental conditions. Therefore, while adenosine-5'-phosphosulfate reductase (APR) and sulfite reductase (SiR) share control under standard laboratory conditions, APR takes over an even more dominant role under sulfur starvation conditions.

Role of APS reductase in biogeochemical sulfur isotope fractionation

Sulfur isotope fractionation resulting from microbial sulfate reduction (MSR) provides some of the earliest evidence of life, and secular variations in fractionation values reflect changes in biogeochemical cycles. Here we determine the sulfur isotope effect of the enzyme adenosine phosphosulfate reductase (Apr), which is present in all known organisms conducting MSR and catalyzes the first reductive step in the pathway and reinterpret the sedimentary sulfur isotope record over geological time. Small fractionations may be attributed to low sulfate concentrations and/or high respiration rates, whereas fractionations greater than that of Apr require a low chemical potential at that metabolic step. Since Archean sediments lack fractionation exceeding the Apr value of 20‰, they are indicative of sulfate reducers having had access to ample electron donors to drive their metabolisms. Large fractionations in post-Archean sediments are congruent with a decline of favorable electron donors as aerobic and other high potential metabolic competitors evolved.