Herbicide Detoxification Research Articles

Measuring Rates of Herbicide Metabolism in Dicot Weeds with an Excised Leaf Assay

In order to isolate and accurately determine rates of herbicide metabolism in an obligate-outcrossing dicot weed, waterhemp (Amaranthus tuberculatus), we developed an excised leaf assay combined with a vegetative cloning strategy to normalize herbicide uptake and remove translocation as contributing factors in herbicide-resistant (R) and –sensitive (S) waterhemp populations. Biokinetic analyses of organic pesticides in plants typically include the determination of uptake, translocation (delivery to the target site), metabolic fate, and interactions with the target site. Herbicide metabolism is an important parameter to measure in herbicide-resistant weeds and herbicide-tolerant crops, and is typically accomplished with whole-plant tests using radiolabeled herbicides. However, one difficulty with interpreting biokinetic parameters derived from whole-plant methods is that translocation is often affected by rates of herbicide metabolism, since polar metabolites are usually not mobile within the plant following herbicide detoxification reactions. Advantages of the protocol described in this manuscript include reproducible, accurate, and rapid determination of herbicide degradation rates in R and S populations, a substantial decrease in the amount of radiolabeled herbicide consumed, a large reduction in radiolabeled plant materials requiring further handling and disposal, and the ability to perform radiolabeled herbicide experiments in the lab or growth chamber instead of a greenhouse. As herbicide resistance continues to develop and spread in dicot weed populations worldwide, the excised leaf assay method developed and described herein will provide an invaluable technique for investigating non-target site-based resistance due to enhanced rates of herbicide metabolism and detoxification.

Measuring Rates of Herbicide Metabolism in Dicot Weeds with an Excised Leaf Assay.

In order to isolate and accurately determine rates of herbicide metabolism in an obligate-outcrossing dicot weed, waterhemp (Amaranthus tuberculatus), we developed an excised leaf assay combined with a vegetative cloning strategy to normalize herbicide uptake and remove translocation as contributing factors in herbicide-resistant (R) and -sensitive (S) waterhemp populations. Biokinetic analyses of organic pesticides in plants typically include the determination of uptake, translocation (delivery to the target site), metabolic fate, and interactions with the target site. Herbicide metabolism is an important parameter to measure in herbicide-resistant weeds and herbicide-tolerant crops, and is typically accomplished with whole-plant tests using radiolabeled herbicides. However, one difficulty with interpreting biokinetic parameters derived from whole-plant methods is that translocation is often affected by rates of herbicide metabolism, since polar metabolites are usually not mobile within the plant following herbicide detoxification reactions. Advantages of the protocol described in this manuscript include reproducible, accurate, and rapid determination of herbicide degradation rates in R and S populations, a substantial decrease in the amount of radiolabeled herbicide consumed, a large reduction in radiolabeled plant materials requiring further handling and disposal, and the ability to perform radiolabeled herbicide experiments in the lab or growth chamber instead of a greenhouse. As herbicide resistance continues to develop and spread in dicot weed populations worldwide, the excised leaf assay method developed and described herein will provide an invaluable technique for investigating non-target site-based resistance due to enhanced rates of herbicide metabolism and detoxification.

Leaf proteome analysis provides insights into the molecular mechanisms of bentazon detoxification in rice

Bentazon is a widely used herbicide that selectively removes broad-leaf weeds by competing with plastoquinone for the binding site in the D1 protein and interrupting the PET (photosynthetic electron transfer) chain. However, monocotyledonous plants, such as rice, show strong resistance to bentazon due to CYP81A6 induction, which results in herbicide detoxification. Here, we confirmed that rice was sensitive to bentazon treatment during the initial exposure period, in which bentazon rapidly inhibited photosynthesis efficiency and electron transfer, based on results of chlorophyll fluorescence analysis. In order to gain a comprehensive, pathway-oriented, mechanistic understanding of the effects directly induced by bentazon, we employed 2D-DIGE (two-dimensional difference gel electrophoresis) to analyze the leaf proteome after 8h of bentazon treatment coupled with individual protein identification by MALDI-TOF (Matrix assisted laser desorption/ionization-time of flight) MS/MS. Proteomic analyses revealed that bentazon induced the relative upregulation or downregulation of 30 and 71 proteins (by 1.5-fold or more, p<0.05), respectively. The pathways involved include photosynthesis processes, carbohydrate metabolism, antioxidant systems, and DNA stabilization and protein folding. Protein analysis data revealed that bentazon primarily suppressed photosynthesis processes, and showed inhibitory effects on carbohydrate metabolism and ATP synthesis, whereas several stress response proteins were induced in response to bentazon. Importantly, we identified a 519kD protein containing two histidine kinase-like ATPase domains and a C3HC4 RING type zinc finger domain which may function as a transcript factor to drive expression of detoxification genes such as CYP81A6, leading to bentazon tolerance. This study identifies, for the first time, a candidate transcription factor that could up-regulate CYP81A6 expression, and provides a foundation for further research to advance our knowledge of mechanisms of bentazon resistance in rice.

Momilactone Sensitive Proteins in Arabidopsis thaliana

The labdane-related diterpenoid, momilactone B has potent growth inhibitory activity and was demonstrated to play a particularly critical role in the allelopathy of rice (Oryza sativa L.). However, there is limited information available about the mode of action of momilactone B on the growth inhibition. The present research describes the effects of momilactone B on protein expression in the early development of Arabidopsis thaliana seedling, which was determined by two-dimensional electrophoresis and MALDI-TOFMS. Momilactone B inhibited the accumulation of subtilisin-like serine protease, amyrin synthase LUP2, β-glucosidase and malate synthase at 1 h after the momilactone application. Those proteins are involved in the metabolic turnover and the production of intermediates needed for cell structures resulting in plant growth and development. Momilactone B also inhibited the breakdown of cruciferin 2, which is essential for seed germination and seedling growth to construct cell structures. Momilactone B induced the accumulation of translationally controlled tumor protein, glutathione S-transferase and 1-cysteine peroxiredoxin 1. These proteins are involved in stress responses and increased stress tolerance. In addition, glutathione S-transferase has the activity of herbicide detoxification and 1-cysteine peroxiredoxin 1 has inhibitory activity for seed germination under unfavorable conditions. The present research suggests that momilactone B may inhibit the seedling growth by the inhibition of the metabolic turnover and the production of intermediates for cell structures. In addition, momilactone induced proteins associated with plant defense responses.

Plant Glutathione Transferases: Structure, Antioxidant Catalytic Function and in planta Protective Role in Biotic and Abiotic Stress

Plant cytosolic glutathione transferases (GSTs) belong to an ancient enzyme superfamily with multiple and diverse functions which are important in counteracting biotic and abiotic stress. GSTs catalyze the conjugation of xenobiotics and endogenous electrophilic compounds with glutathione (GSH), leading to their detoxification. GSTs not only catalyze detoxification reactions but they are also involved in GSH-dependent isomerization reactions, in GSH-dependent reduction of organic hydroperoxides, biosynthesis of secondary metabolites, and exhibit thioltransferase and dehydroascorbate reductase activity. The applications of ‘omics’ technologies have allowed the classification of GSTs and the study of their evolution and sequence diversity, while enzymology has provided powerful insights into their catalytic role. This review focuses on plant GSTs, and attempts to give an overview of the new insights into their catalytic function and biological role in biotic and abiotic stress tolerance mechanisms in plants. Keywords: Abiotic stress, biotic stress, glutathione transferase, herbicide detoxification.

A glutathione s-transferase confers herbicide tolerance in rice

Plant glutathione S-transferases (GSTs) have been a focus of attention due to their role in herbicide detoxification. OsGSTL2 is a glutathione S-transferase, lambda class gene from rice (Oryza sativa L.). Transgenic rice plants over-expressing OsGSTL2 were generated from rice calli by the use of an Agrobacterium transformation system, and were screened by a combination of hygromycin resistance, PCR and Southern blot analysis. In the vegetative tissues of transgenic rice plants, the over-expression of OsGSTL2 not only increased levels of OsGSTL2 transcripts, but also GST and GPX expression, while reduced superoxide. Transgenic rice plants also showed higher tolerance to glyphosate and chlorsulfuron, which often contaminate agricultural fields. The findings demonstrate the detoxification role of OsGSTL2 in the growth and development of rice plants. It should be possible to apply the present results to crops for developing herbicide tolerance and for limiting herbicide contamination in the food chain.

Cyperus difformis evolves resistance to propanil

Cyperus difformis L. is one of the worst weeds of rice world-wide and has evolved resistance to acetolactate synthase (ALS)-inhibiting herbicides in rice fields of California. Propanil use was intensified to control the widespread resistant biotypes. Rice growers have recently experienced poor control, suggesting resistance to this photosystem II-inhibiting herbicide may have evolved in C. difformis populations. The objectives of this study were to detect the presence of propanil resistance, to establish resistance levels, and to investigate involvement of enhanced herbicide detoxification as mechanism of resistance through the use of metabolic inhibitors. Four C. difformis populations collected in rice fields from the Sacramento Valley of California were confirmed resistant to propanil. This is the first case of such resistance outside the Poaceae and the first time C. difformis exhibits resistance to an herbicide mechanism of action other than ALS inhibition. Carbaryl and malathion applied individually in mixture with propanil had minor effects on herbicide toxicity suggesting metabolic detoxification was not a resistance mechanism. A resistant biotype produced more than 80% biomass after a propanil (6.7 kg a.i. ha−1) and carbaryl (1.9 kg a.i. ha−1) or propanil and malathion (1.0 kg a.i. ha−1) treatment compared to <20% by a susceptible biotype, suggesting substantial resistance still persisted in spite of insecticide addition. Propanil-resistant plants were cross-resistant to bensulfuron-methyl, imazosulfuron, halosulfuron-methyl and penoxsulam, but susceptible to carfentrazone. The loss of propanil to control this important weed of rice underscores the fragility of herbicide-based weed control in monoculture rice. Integrated weed management approaches to decrease herbicide selection pressure are needed to mitigate the evolution of multiple-herbicide resistance in C. difformis of California rice.

Characterization of a novel cold-adapted phosphinothricin N-acetyltransferase from the marine bacterium Rhodococcus sp. strain YM12

Abstract Phosphinothricin (PPT) is a kind of non-selective, environmentally friendly herbicide. PPT-tolerance genes are vital in both plant biotechnology as selectable markers and the development of transgenic herbicide-resistant crops. However, there are no other well-identified and commercially available PPT-resistance genes for use in plant genetic engineering besides two PPT N -acetyltransferase genes, which known as pat and bar derived from Streptomyces sp. Here, we isolated a novel PPT N -acetyltransferase gene from PPT-resistant marine bacteria, Rhodococcus sp. strain YM12 . The gene, designated as RePAT , encoded a protein (RePAT) of 162 amino acids, which showed 37% identity with that of PAT proteins. Key kinetic constants of RePAT were determined ( K m = 0.076 mM, K cat = 131 min −1 ) using PPT as a substrate, the enzyme retained considerable activity at pH 8.0 and had an optimum temperature of 35 °C. Interestingly, it possessed over 50% of its maximal activity at temperature conditions between 0 and 10 °C, suggesting that this enzyme is able to protect crop against PPT injury in cold environment. These results illustrated that RePAT could be a new resource for herbicide detoxification by transgenic crops.

Biochemical functions of the glutathione transferase supergene family of Larix kaempferi

Glutathione transferases (GSTs), which are ubiquitous in plants, play a major role in the detoxification of xenobiotics and oxidative stress metabolism. Due to their role in herbicide detoxification, previous studies of plant GSTs have mainly focused on agricultural plants. In contrast, functional information regarding gymnosperm GSTs is scarce. In this study, we cloned 27 full-length GST genes from the deciduous conifer Larix kaempferi, which is widely distributed across the cooler regions of the northern hemisphere. As with the angiosperm GST gene family, Larix GSTs are divided into eight classes, and tau class GSTs are the most numerous. Compared to the other seven classes of GSTs, Larix tau GST genes show substantially more variation in their expression patterns. The purified Larix GST proteins showed different substrate specificities, substrate activities, and kinetic characteristics. The pH and temperature profiles of purified Larix GST proteins showed broad optimum pH and temperature ranges for enzymatic activity, suggesting that Larix GSTs have evolutionary adaptations to various adverse environments. Taken together, this study provides comprehensive insight into the gymnosperm GST gene family.

A bioconjugated design for amino acid-modified mesoporous silicas as effective adsorbents for toxic chemicals

A general synthetic method for functionalization of mesoporous silica with amino acid has been developed. The carboxylic acid functionalized SBA-15 was conjugated with l-phenylalanine (Phe) and l-tryptophan (Trp) to obtain nontoxic amino acid-conjugated functionalized mesoporous silica materials. The materials were used as adsorbents for the removal of the herbicide Paraquat (PQ) and its analog, ethyl viologen dibromide (EVB) from aqueous solutions. In comparison to the commercially available activated carbon adsorbents, the silica-based adsorbents prepared in this study exhibited relatively higher PQ removal efficiency in aqueous solutions at room temperature and pH 7.0. The silica-based adsorbents, pendant with amino acid moieties exhibited greater adsorption capacities toward PQ and EVB than the analogs but without the amino acid moiety, suggesting that there is a benefit for the enhanced π–π interaction between the aromatic groups of the conjugated amino acid moieties and the adsorbate. This bioconjugated method developed here provides a promising new tool to synthesize new materials for detoxification of herbicides in clinical trials.

Identification and expression pattern of a glutathione S‐transferase in Echinochloa crus‐galli

SummaryPlant glutathione S‐transferase (GST) forms a major part of the herbicide detoxification enzyme network in plants. A GST cDNA was isolated from Echinochloa crus‐galli and characterised. The gene, designated EcGST1 (E. crus‐galli GeneBank no: JX518596), has a 684 bp open reading frame predicted to encode a 25 kD protein. Sequence alignment showed that EcGST1 is a GST homologue. Its expression in response to quinclorac treatment was monitored in seedlings (leaves and roots) and adult plants (leaves, roots, stems and seeds) of quinclorac‐resistant (R) and susceptible (S) biotypes of E. crus‐galli. EcGST1 expression was 1.5–3 times greater in the R plants than in the S plants. However, after exposure to quinclorac, the difference in the expression levels of EcGST1 in R plants, compared with S plants, increased to a ratio of 6–10. Enhanced EcGST1 levels should enable greater quinclorac detoxification following quinclorac stimulation in R plants. GST‐based metabolism may be partially responsible for resistance to quinclorac in E. crus‐galli. The results suggest a new resistance mechanism for this R biotype in Chinese rice fields.

English

Plant glutathione S-transferases (GSTs) have been a focus of attention because of their role in herbicide detoxification.&nbsp;OsGSTL2&nbsp;is a glutathione S-transferase, lambda class gene from rice. Transgenic rice&nbsp;plants&nbsp;overexpressing OsGSTL2 were generated from rice calli by the use of an&nbsp;Agrobacteriumtransformation system. The transgenic rice&nbsp;plants&nbsp;were screened by a combination of hygromycin resistance, polymerase chain reaction (PCR) and Southern blot analysis. Transgenic rice plants overexpressing&nbsp;OsGSTL2&nbsp;gene showed higher levels of&nbsp;OsGSTL2&nbsp;transcripts in the absence of any treatment compared to non-transformed rice plants. In the vegetative tissues of transgenic rice plants, the overexpression of the&nbsp;OsGSTL2&nbsp;increased levels of GST and glutathione peroxidase (GPX) activities, and reduced content of superoxide. Transgenic rice plants also had higher tolerance to glyphosate and chlorsulfuron, which often contaminates agricultural fields. These findings demonstrate the detoxification role of OsGSTL2 in the growth and development of rice plants.&nbsp;It should be possible to apply the present results to crop&nbsp;plants&nbsp;for developing&nbsp;herbicides&nbsp;tolerance and in limiting&nbsp;herbicides&nbsp;availability in the food chain. Key words: Glutathione S-transferase, transgenic rice, overexpression, herbicide resistance.

Salicylic acid induced alleviation of oxidative stress caused by clethodim in maize (Zea mays L.) leaves

Salicylic acid is used for regulation of oxidative stress in plants subjected to unfavorable environmental conditions. Application of herbicides for the purpose of weed killing can affect not only the weeds but also the main crop as well. Many herbicides have the ability to cause oxidative stress and further degradation of cell components. In this work, SA was used to alleviate the oxidative stress caused in response to clethodim herbicide in maize leaves. The results demonstrated that, spraying of clethodim caused yellowing of leaves and sometimes browning or drying of leaf tips with high clethodim doses. Contrary, leaves showed no injuries when treated with 1mM SA 3days prior clethodim application. Elevated amounts of H2O2 and MDA were detected in clethodim treated leaves compared with control indicating ROS formation and lipid peroxidation. Excessive ROS formation led to oxidative stress which followed by degradation of membranous structures. In SA treated leaves, the contents of H2O2 and MDA were more or less similar to the corresponding controls. A change in the antioxidant enzymes activities due to clethodim and SA treatment was noticed. For example, the activities of POD and APX were induced while SOD and CAT were more or less reduced in response to clethodim. SA treatment prior clethodim application could induce POD but inhibit CAT. Moreover, SOD and APX activities were adjusted to be similar to those of the control. Another mechanism of SA regulation of the oxidative stress occurred through the formation of antioxidants in the form of phenolic compounds. For that, spraying SA with or without clethodim could accumulate phenolic compounds greatly. The DPPH free radical scavenging assay for leaf extracts had confirmed a change in antioxidant status. Furthermore, SA could enhance accumulation of total proteins and free amino acids in clethodim and SA treated leaves compared with the control. This work was to provide evidence for the ability of SA to regulate clethodim herbicide detoxification through regulation of the antioxidant status of maize leaf cells.

Catalytic and structural diversity of the fluazifop-inducible glutathione transferases from Phaseolus vulgaris

Plant glutathione transferases (GSTs) comprise a large family of inducible enzymes that play important roles in stress tolerance and herbicide detoxification. Treatment of Phaseolus vulgaris leaves with the aryloxyphenoxypropionic herbicide fluazifop-p-butyl resulted in induction of GST activities. Three inducible GST isoenzymes were identified and separated by affinity chromatography. Their full-length cDNAs with complete open reading frame were isolated using RACE-RT and information from N-terminal amino acid sequences. Analysis of the cDNA clones showed that the deduced amino acid sequences share high homology with GSTs that belong to phi and tau classes. The three isoenzymes were expressed in E. coli and their substrate specificity was determined towards 20 different substrates. The results showed that the fluazifop-inducible glutathione transferases from P. vulgaris (PvGSTs) catalyze a broad range of reactions and exhibit quite varied substrate specificity. Molecular modeling and structural analysis was used to identify key structural characteristics and to provide insights into the substrate specificity and the catalytic mechanism of these enzymes. These results provide new insights into catalytic and structural diversity of GSTs and the detoxifying mechanism used by P. vulgaris.

A Plant-specific Tau Class Glutathione S-transferase from Oryza sativa Having Significant Detoxification Activity Towards Chloroacetanilide Herbicides

Glutathione S-transferases (GSTs, EC 2.5.1.18) are enzymes that detoxify endobiotic and xenobiotic compounds by covalently linking glutathione (GSH) to a hydrophobic substrate, forming a less reactive and more polar glutathione S-conjugate. 1 In plants, GSTs detoxify herbicides, organic pollutants, and natural toxins; they also protect cells from a wide range of biotic and abiotic stresses, including pathogen attack, xenobiotic and heavy-metal toxicity, and oxidative stress. 2,3 Plant GSTs are grouped into five classes (phi, zeta, tau, theta and lambda) based on sequence identity, gene organization, and active site residues in the proteins. A further GST-like class, DHAR (proteins with dehydroascorbate reductase activity), was recently reported with members in Arabidopsis, rice and soybean. 2 The function of these plant-specific GSTs is detoxification of herbicides in both crops and weeds. Despite all available research, both the biochemical properties and functions of plant GSTs remain to be elucidated. Rice is the most important crop for human consumption with more than a half of the world’s population utilizing it as an energy source. The entire rice genomes for the japonica and indica subspecies have been sequenced. 4 From these genome sequences, a GST gene homolog (GenBank Accession No. AF309379) was identified from Oryza sativa by homolog searches in the NCBI database. As yet, there have been no reports concerning this gene product. Therefore, we cloned this gene and expressed it in Escherichia coli. The mRNA from Oryza sativa L. cv. Yamahousi was isolated from the cell culture and transcribed into cDNA. Primers for PCR, using the cDNA as a template, were derived from a published cDNA sequence from Oryza sativa L. cv. Japonica. 5 Subsequently, a 702-bp fragment of Oryza sativa cDNA was amplified by PCR, subcloned into the expression vector pET-26b(+) and transformed into the E. coli strain BL21 (DE3). Both DNA strands were sequenced with the final amino acid sequence bearing a difference between the PCR-identified sequence and the earlier cDNAderived sequence (GenBank Accession No. AF309379). The gene whereby guanine was substituted with thymine at position of nucleotide 166 gave an Asn 55 residue instead of Lys 55. The cloned gene was composed of 702-bp encoding for the 233 amino acids and designated OsGSTU3. A multiple alignment of the OsGSTU3 protein with other GSTs is shown in Figure 1. Sequence aliment was performed using the T-Coffee and Boxshade programe (http:// www.ch.embnet.org). GST isoenzymes belonging to the same class showed more than approximately 60% identity in their primary structure, whereas enzymes belonging to phi and theta classes generally had less than 2% sequence identity. 2 Zeta class enzymes also bore less than 24% sequence identity. Based on its gene sequence, OsGSTU3 was presumed to belong to the plant-specific tau class GST. The E. coli BL21 (pET-OsGSTU3) clone was tested for production of recombinant OsGSTU3 protein. The expressed OsGSTU3 protein was purified by S-hexylGSH affinity

English

Glutathione S-transferases are multifunctional proteins involved in diverse intracellular events such as primary and secondary metabolisms, stress metabolism, herbicide detoxification and plant protection against ozone damages, heavy metals and xenobiotics. The plant glutathione S-transferase superfamily have been subdivided into eight classes. Phi, tau, zeta, theta, lambda, dehydroascorbate reductase and tetrachlorohydroquinone dehalogenase classes are soluble and one class is microsomal. Glutathione S-transferases are mostly soluble cytoplasmic enzymes. To date, the crystal structures of over 200 soluble glutathione S-transferases, present in plants, animals and bacteria have been resolved. The structures of glutathione S-transferase influence its function. Phylogenetic analysis suggests that all soluble glutathione S-transferases have arisen from an ancient progenitor gene, through both convergent and divergent pathways. Key words: Glutathione S-transferases (GST), classification, structure, evolution, phylogenetic analysis, xenobiotics.

Tau class glutathione S-transferases candidates from Brassica rapa: Genetic mapping and sequence analysis

Glutathione S-transferases (GSTs) have medically and agriculturally important roles in plants such as herbicide detoxification, responses to biotic and abiotic stress and catalytic reaction of anticancer compounds. The fully sequenced Arabidopsis thaliana and rice (Oryza sativa) genome revealed identification of 52 and 61 members of GSTs, and 28 and 39 of those belong to the Tau class of GSTs (GSTUs), respectively. Based on the sequences of AtGSTUs, 14 BAC clones derived from Brassica rapa that contain similar sequences of AtGSTUs were identified and 17 unique sequences of open reading frames were detected in the 14 BAC clones using blast search and sequence alignment. Those sequences were designated the Tau class candidate of GST derived from B. rapa (BrGSTUs) and it is confirmed that BrGSTUs are also clustered in tandem as it has been known to be common feature in plant. They were mapped on BrR5, BrR7, BrR8, BrR9, or BrR10, and their nucleotide and amino acid sequences were highly similar to those of AtGSTUs. In addition, in silico analysis of BrGSTUs was performed using Korea Brassica Genome Project 24 K (KBGP-24K) oligochip and microarray database for cold, salt, and drought stresses. The results of this study will facilitate further researches for breeding of Chinese cabbage containing medically and agriculturally important traits.

Plant Glutathione Biosynthesis: Diversity in Biochemical Regulation and Reaction Products

In plants, exposure to temperature extremes, heavy metal-contaminated soils, drought, air pollutants, and pathogens results in the generation of reactive oxygen species that alter the intracellular redox environment, which in turn influences signaling pathways and cell fate. As part of their response to these stresses, plants produce glutathione. Glutathione acts as an anti-oxidant by quenching reactive oxygen species, and is involved in the ascorbate–glutathione cycle that eliminates damaging peroxides. Plants also use glutathione for the detoxification of xenobiotics, herbicides, air pollutants (sulfur dioxide and ozone), and toxic heavy metals. Two enzymes catalyze glutathione synthesis: glutamate–cysteine ligase, and glutathione synthetase. Glutathione is a ubiquitous protective compound in plants, but the structural and functional details of the proteins that synthesize it, as well as the potential biochemical mechanisms of their regulation, have only begun to be explored. As discussed here, the core reactions of glutathione synthesis are conserved across various organisms, but plants have diversified both the regulatory mechanisms that control its synthesis and the range of products derived from this pathway. Understanding the molecular basis of glutathione biosynthesis and its regulation will expand our knowledge of this component in the plant stress response network.

Conserved Residues in the Subunit Interface of tau Glutathione S‐transferase Affect Catalytic and Structural Functions

The tau class glutathione S-transferases (GSTs) have important roles in stress tolerance and the detoxification of herbicides in crops and weeds. Structural investigations of a wheat tau GST (TaGSTU4) show two subunit interactions: a hydrogen bond between the Tyr93 and Pro65 from another subunit of the dimer, and two salt bridges between residues Glu78 and side chains of Arg95 and Arg99 in the opposite subunit. By investigating enzyme activities, kinetic parameters and structural characterizations, this study showed the following results: (i) the hydrogen bond interaction between the Tyr93 and Pro65 was not essential for dimerization, but contributed to the enzyme's catalytic activity, thermal stability and affinity towards substrates glutathione and 1-chloro-2, 4-dinitrobenzene; and (ii) two salt bridges mainly contributed to the protein structure stability and catalysis. The results of this study form a structural and functional basis for rational design of more selective and environmentally friendly herbicides.

Agronomic and environmental implications of enhanceds-triazine degradation

Novel catabolic pathways enabling rapid detoxification of s-triazine herbicides have been elucidated and detected at a growing number of locations. The genes responsible for s-triazine mineralization, i.e. atzABCDEF and trzNDF, occur in at least four bacterial phyla and are implicated in the development of enhanced degradation in agricultural soils from all continents except Antarctica. Enhanced degradation occurs in at least nine crops and six crop rotation systems that rely on s-triazine herbicides for weed control, and, with the exception of acidic soil conditions and s-triazine application frequency, adaptation of the microbial population is independent of soil physiochemical properties and cultural management practices. From an agronomic perspective, residual weed control could be reduced tenfold in s-triazine-adapted relative to non-adapted soils. From an environmental standpoint, the off-site loss of total s-triazine residues could be overestimated 13-fold in adapted soils if altered persistence estimates and metabolic pathways are not reflected in fate and transport models. Empirical models requiring soil pH and s-triazine use history as input parameters predict atrazine persistence more accurately than historical estimates, thereby allowing practitioners to adjust weed control strategies and model input values when warranted.