Tripyrasulfone is a novel herbicide post-emergence applied in paddy fields. In this study, tripyrasulfone phytotoxicity and its mode of action were investigated. Within 3-7 days after treatment (DAT), tripyrasulfone caused strong bleaching symptoms on newly developed leaves of followed by necrosis prior to death within 14 DAT. By investigating pigment composition, photosynthetic activity and energy dissipation of treated with tripyrasulfone, the accumulation of phytoene and significant decreases in total carotenoids were observed; the photosystem II complex (PSII) reaction center and PSII-PSI electron transport chain were damaged; and the non-photochemical energy quenching and reactive oxygen species were significantly increased. Based on the reversion of bleaching symptoms in treated by the addition of homogentisic acid, it was hypothesized that tripyrasulfone blocks the biosynthesis of HGA, possibly by the inhibition of 4-hydroxyphenylpyruvate dioxygenase (HPPD). However, based on its chemical structure, tripyrasulfone may tend to be hydrolyzed in plants. Indeed, the hydrolyzed tripyrasulfone (HDT) inhibited the activity of HPPD from produced by, which was approximately 6 times less effective than mesotrione. Molecular docking showed that the HDT formed a stable bidentate interaction with the active center Fe chelation of HPPD.

Carotenoids play an important role in many physiological processes in plants and the phytoene desaturase gene (PDS3) encodes one of the important enzymes in the carotenoid biosynthesis pathway. Here we report the identification and analysis of a T-DNA insertion mutant of PDS3 gene. Functional complementation confirmed that both the albino and dwarf phenotypes of the pds3 mutant resulted from functional disruption of the PDS3 gene. Chloroplast development was arrested at the proplastid stage in the pds3 mutant. Further analysis showed that high level of phytoene was accumulated in the pds3 mutant. Addition of exogenous GA(3) could partially rescue the dwarf phenotype, suggesting that the dwarf phenotype of the pds3 mutant might be due to GA deficiency. Microarray and RT-PCR analysis showed that disrupting PDS3 gene resulted in gene expression changes involved in at least 20 metabolic pathways, including the inhibition of many genes in carotenoid, chlorophyll, and GA biosynthesis pathways. Our data suggest that the accumulated phytoene in the pds3 mutant might play an important role in certain negative feedbacks to affect gene expression of diverse cellular pathways.

Field and greenhouse experiments were conducted in Nebraska to (1) confirm the 4-hydroxyphenylpyruvate dioxygenase (HPPD)-inhibiting resistant-waterhemp biotype (HPPD-RW) by quantifying the resistance levels in dose-response studies, and (2) to evaluate efficacy of PRE-only, POST-only, and PRE followed by POST herbicide programs for control of HPPD-RW in corn. Greenhouse dose-response studies confirmed that the suspected waterhemp biotype in Nebraska has evolved resistance to HPPD-inhibiting herbicides with a 2- to 18-fold resistance depending upon the type of HPPD-inhibiting herbicide being sprayed. Under field conditions, at 56 d after treatment, ≥90% control of the HPPD-RW was achieved with PRE-applied mesotrione/atrazine/S-metolachlor+acetochlor, pyroxasulfone (180 and 270 g ai ha−1), pyroxasulfone/fluthiacet-methyl/atrazine, and pyroxasulfone+saflufenacil+atrazine. Among POST-only herbicide programs, glyphosate, a premix of mesotrione/atrazine tank-mixed with diflufenzopyr/dicamba, or metribuzin, or glufosinate provided ≥92% HPPD-RW control. Herbicide combinations of different effective sites of action in mixtures provided ≥86% HPPD-RW control in PRE followed by POST herbicide programs. It is concluded that the suspected waterhemp biotype is resistant to HPPD-inhibiting herbicides and alternative herbicide programs are available for effective control in corn. The occurrence of HPPD-RW in Nebraska is significant because it limits the effectiveness of HPPD-inhibiting herbicides.

Safeners extend the application of existing herbicides by selectively enhancing tolerance in large-grained cereal crops. While their activity is linked to enhanced herbicide metabolism, their exact mode of action and reasons for their crop specificity have yet to be determined. In this study, we investigated the selectivity of the recently developed sulfonamide safener cyprosulfamide (CSA) in maize (Zea mays L.) and wheat (Triticum aestivum), focusing on its uptake, distribution and metabolism in the two species.CSA protected maize, but not wheat, from injury by thiencarbazone-methyl (TCM). This correlated with the selective enhanced detoxification of the herbicide in maize. CSA underwent more rapid metabolism in maize than in wheat, with the formation of a specific hydroxylated metabolite correlating with safening. Studies with the nsf1 mutant sweetcorn line showed that the hydroxylation of CSA was partly mediated by the cytochrome P450 CYP81A9. However, primary metabolites of CSA were chemically synthesised and tested for their ability to safen TCM in maize but when tested were inactive as safeners.The results of this study suggest that the protection against TCM injury by CSA is linked to enhanced herbicide metabolism. This selective activity is due to the specific recognition of parent CSA in maize but not in wheat. Subsequent rapid oxidative metabolism of CSA led to its inactivation, demonstrating that cytochrome P450s regulate the activity of safeners as well as herbicides. © 2020 Society of Chemical Industry.© 2020 Society of Chemical Industry.

Detoxification (detox) plays a major role in pesticide action and resistance. The mechanisms involved are sometimes part of the discovery and development process in seeking new biochemical targets and metabolic pathways. Genetically modified and chemical-safener-modified crops are a marked exception and often involve herbicide detox by design to achieve the required crop tolerance. This perspective evaluates the role of detox by design or chance and target-site-based selectivity in insecticide, herbicide, and fungicide action and human health and environmental effects.

The closely related sulphonamide safeners, metcamifen and cyprosulfamide, were tested for their ability to protect rice from clodinafop-propargyl, a herbicide normally used in wheat. While demonstrating that both compounds were equally bioavailable in planta, only metcamifen prevented clodinafop from damaging seedlings, and this was associated with the enhanced detoxification of the herbicide. Transcriptome studies in rice cultures demonstrated that whereas cyprosulfamide had a negligible effect on gene expression over a 4 h exposure, metcamifen perturbed the abundance of 590 transcripts. Changes in gene expression with metcamifen could be divided into three phases, corresponding to inductions occurring over 30 min, 1.5 h and 4 h. The first phase of gene induction was dominated by transcription factors and proteins of unknown function, the second by genes involved in herbicide detoxification, while the third was linked to cellular homeostasis. Analysis of the inducible genes suggested that safening elicited similar gene families to those associated with specific biotic and abiotic stresses, notably those elicited by abscisic acid, salicylic acid, and methyl jasmonate. Subsequent experiments with safener biomarker genes induced in phase 1 and 2 in rice cell cultures provided further evidence of similarities in signalling processes elicited by metcamifen and salicylic acid.

Several grass and broadleaf weed species around the world have evolved multiple-herbicide resistance at alarmingly increasing rates. Research on the biochemical and molecular resistance mechanisms of multiple-resistant weed populations indicate a prevalence of herbicide metabolism catalyzed by enzyme systems such as cytochrome P450 monooxygenases and glutathioneS-transferases and, to a lesser extent, by glucosyl transferases. A symposium was conducted to gain an understanding of the current state of research on metabolic resistance mechanisms in weed species that pose major management problems around the world. These topics, as well as future directions of investigations that were identified in the symposium, are summarized herein. In addition, the latest information on selected topics such as the role of safeners in inducing crop tolerance to herbicides, selectivity to clomazone, glyphosate metabolism in crops and weeds, and bioactivation of natural molecules is reviewed.

Safeners have been widely used to reduce phytotoxicity to crops, thus serving as an alternative weed control strategy. Benoxacor and fenclorim safeners have the potential to protect plants from herbicide phytotoxicity by increasing glutathione S-transferase (GST) activity within the plant. The study aimed to evaluate the safening effect of benoxacor and fenclorim on tomato against selected herbicides applied POST. The experiment was conducted in a greenhouse in a completely randomized designed with four replications in a 9 × 3 factorial scheme, where Factor A consisted of eight herbicides including a nontreated control, and Factor B consisted of two safeners including a nontreated control. The herbicide treatments were sulfentrazone (0.220 kg ai ha−1), fomesafen (0.280 kg ai ha−1), flumioxazin (0.070 kg ai ha−1), linuron (1.200 kg ai ha−1), metribuzin (0.840 kg ai ha−1), pyroxasulfone (0.220 kg ai ha−1), and bicyclopyrone (0.040 kg ai ha−1). Safener treatments consisted of benoxacor (0.67 g L−1) and fenclorim (10 µM). Tomato seeds were immersed in safener solution before sowing and herbicides were applied when tomato plants were at the 3-leaf stage, or 25 days after sowing. Visible injury was scored at 3, 7, 14, and 21 d after application (DAA), and shoot biomass was recorded 21 DAA. Seed treatment with fenclorim reduced injury caused by imazamox and bicyclopyrone by 5.5 and 1.3 times, respectively, whereas benoxacor reduced the injury from bicyclopyrone 1.3 times. In addition, tomato plants pretreated with fenclorim showed a lesser reduction in biomass after application of imazamox, fomesafen, and metribuzin, whereas plants pretreated with benoxacor showed lesser biomass reduction after metribuzin application. Thus, the use of safeners promotes greater crop selectivity, allowing the application of herbicides with different mechanisms of action on the crop.

Herbicide safeners are a series of agrochemicals that can selectively protect crop plants from herbicide injury without affecting herbicidal efficacy. Understanding mechanisms by which safeners act is significant for discovery of novel types. Safeners primarily alleviate herbicide phytotoxicity to crop plants via several actions: (i) enhancing metabolism of herbicides in crops; (ii) affecting absorption and transportation of herbicides in crops; (iii) competitively binding to herbicide target sites; and (iv) affecting activity of target enzymes. This review describes recent advances in the action mechanisms of safeners, analyzes existing problems, anticipates the future direction of studies of modes of action of safeners, and prospects potential strategies to design safeners related to their reported mechanisms. The aim of this paper is to provide insight into mechanisms of safeners and give tips for development of new safeners.

为了解安全剂双苯恶唑酸缓解除草剂精恶唑禾草灵对水稻药害的效果，通过温室水培和盆栽法研究双苯恶唑酸对水稻精恶唑禾草灵药害的解毒效果，并对其解毒机制进行初步探讨。结果表明：水稻萌发期，粳稻和籼稻在0.25、0.5、1 μg/mL精恶唑禾草灵处理下，解毒效果最好的安全剂添加比例分别为1∶1、1∶5、1∶5，粳稻株高、胚根长的抑制率比精恶唑禾草灵单独使用分别降低18.07、37.50，26.56、34.94，41.30、37.50个百分点，籼稻株高、胚根长的抑制率分别降低23.31、42.33，27.22、34.91，46.76、31.14个百分点；水稻两至三叶期，粳稻和籼稻在25、100 μg/mL精恶唑禾草灵处理下，谷胱甘肽-S-转移酶(GST)和乙酰辅酶A羧化酶(ACCase)活性最高的安全剂添加比例均为1∶1，此时粳稻的GST活性和ACCase活性比精恶唑禾草灵单独使用分别增加42.21%和65.63%、46.72%和140.43%，籼稻的GST活性和ACCase活性比精恶唑禾草灵单独使用分别增加52.18%和105.71%、55.55%和169.52%。安全剂双苯恶唑酸提高了精恶唑禾草灵处理后水稻幼苗中GST、ACCase活性，减轻了精恶唑禾草灵对水稻的伤害，为水稻生产中通过添加双苯恶唑酸降低精恶唑禾草灵对水稻的药害提供了依据。

The expression of glutathione S-transferase (GST) activity in wheat and maize shoots was investigated in response to treatments with the herbicide safeners benoxacor, cloquintocet-mexyl, fenchlorazole-ethyl, fenclorim, fluxofenim and oxabetrinil. These safeners significantly enhanced the GST activity towards 1-chloro-2,4-dinitrobenzene (CDNB) as a 'standard' substrate, with the exception of oxabetrinil in maize. The enhancements of GST (CDNB) activity were found to be concomitant with increases in V(max) (the reaction rate when the enzyme is fully saturated by the substrate) in wheat following cloquintocet-mexyl and fenchlorazole-ethyl treatments, and in maize following fenchlorazole-ethyl treatment. Otherwise, decreases in V(max) were observed in wheat and maize following fenclorim and fluxofenim treatments. With the exception of oxabetrinil, all the safeners significantly reduced the apparent K(M) (the substrate concentration required for 50% of maximum GST activity) of both wheat and maize GST. The V(max) and K(M) variations following safener treatments are discussed in terms of an increased expression of GST enzymes and an increased affinity for the CDNB substrate. The activity of wheat and maize GST was also assayed towards butachlor and terbuthylazine respectively; the results indicate the ability of cloquintocet-mexyl, fenchlorazole-ethyl and fluxofenim to enhance the enzyme activity in wheat and of benoxacor and fenchlorazole-ethyl to do so in maize.

To support the key role of glutathione (GSH) in the mechanisms of tolerance and accumulation of arsenic in plants, this work examines the impact of several effectors of GSH synthesis or action in the response of maize (Zea mays L.) to arsenic. Maize was exposed in hydroponics to iso-toxic rates of 150 μM arsenate or 75 μM arsenite for 9 days and GSH effectors, flurazole (an herbicide safener), l-buthionine-sulfoximine (BSO, a known inhibitor of GSH biosynthesis), and dimercaptosuccinate (DMS) and dimercaptopropanesulfonate (DMPS) (two thiols able to displace GSH from arsenite-GSH complexes) were assayed. The main responses of plants to arsenic exposure consisted of a biomass reduction (fresh weight basis) of about 50%, an increase of non-protein thiol (NPTs) levels (especially in the GSH precursor γ-glutamylcysteine and the phytochelatins PC₂ and PC₃) in roots, with little effect in shoots, and an accumulation of between 600 and 1000 ppm of As (dry weight basis) in roots with very little translocation to shoots. Growth inhibition caused by arsenic was partially or completely reversed in plants co-treated with flurazole and arsenate or arsenite, respectively, highly exacerbated in plants co-treated with BSO, and not modified in plants co-treated with DMS or DMPS. These responses correlated well with an increase of both NPTs levels in roots and glutathione transferase activity in roots and shoots due to flurazole treatment, the decrease of NPTs levels in roots caused by BSO and the lack of effect on NPT levels caused by both DMS and DMPS. Regarding to arsenic accumulation in roots, it was not modified by flurazole, highly reduced by BSO, and increased between 2.5- and 4.0-fold by DMS and DMPS. Therefore, tolerance and accumulation of arsenic by maize could be manipulated pharmacologically by chemical effectors of GSH.Copyright © 2012 Elsevier GmbH. All rights reserved.

Safeners are agrochemicals used in agriculture to protect crops from herbicide injuries. They act by stimulating herbicide metabolism. As graminaceous plants, to cope with iron (Fe) deficiency, activate sulfur (S) metabolism and release huge amounts of Fe-chelating compounds, or phytosiderophores (PSs), we investigated, in barley plants (Hordeum vulgare, L.) grown in Fe deficiency, the effects of three safeners on two enzymes of S assimilation, cysteine (Cys) and glutathione (GSH), and PS release. Finally, we monitored the root Fe content in plants treated with the most effective safener.Generally, all the safeners activated S metabolism and increased Cys and GSH contents. In addition, the safened plants excreted higher levels of PSs. Given that mefenpyr-diethyl (Mef) was the most effective in causing these effects, we assessed the Fe concentration in Mef-treated barley and found higher Fe levels than those in untreated plants.The three safeners, in different ways but specifically, activated S reductive metabolism and regulated Cys and GSH contents, PS release rate and Fe content (Mef-treated barley). The results of this research provide new indications of the biochemical and physiological mechanisms involved in the safening action. © 2016 Society of Chemical Industry.© 2016 Society of Chemical Industry.

A series of novel substituted oxazole isoxazole carboxamides derivatives were designed on the basis of active subunit combination. Forty-four novel compounds were synthesized by an efficient one-pot procedure under microwave irradiation. The bioactivity was evaluated as herbicide safener against the injury of chlorsulfuron. It was found that most of the synthesized compounds displayed remarkable protection against chlorsulfuron via enhanced glutathione content and glutathione S transferase activity. Especially compound I-11 exhibited better bioactivity than the safeners isoxadifen-ethyl and R-28725. Molecular docking simulations suggested that the target compounds could compete with chlorsulfuron in the active site of acetolactate synthase, which could explain the protective effects of safeners. The present work demonstrates that the target compounds containing oxazole isoxazole groups could be considered as potential candidates for developing novel safeners in the future.Copyright © 2019 Elsevier Inc. All rights reserved.

Herbicide safeners increase herbicide tolerance in cereals but not in dicotyledenous crops. The reason(s) for this difference in safening is unknown. However, safener-induced protection in cereals is associated with increased expression of herbicide detoxifying enzymes, including glutathione S-transferases (GSTs). Treatment of Arabidopsis seedlings growing in liquid medium with various safeners similarly resulted in enhanced GST activities toward a range of xenobiotics with benoxacor, fenclorim, and fluxofenim being the most effective. Safeners also increased the tripeptide glutathione content of Arabidopsis seedlings. However, treatment of Arabidopsis plants with safeners had no effect on the tolerance of seedlings to chloroacetanilide herbicides. Each safener produced a distinct profile of enhanced GST activity toward different substrates suggesting a differential induction of distinct isoenzymes. This was confirmed by analysis of affinity-purified GST subunits by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AtGSTU19, a tau class GST, was identified as a dominant polypeptide in all samples. When AtGSTU19 was expressed in Escherichia coli, the recombinant enzyme was highly active toward 1-chloro-2,4-dinitrobenzene, as well as chloroacetanilide herbicides. Immunoblot analysis confirmed that AtGSTU19 was induced in response to several safeners. Differential induction of tau GSTs, as well as members of the phi and theta classes by safeners, was demonstrated by RNA-blot analysis. These results indicate that, although Arabidopsis may not be protected from herbicide injury by safeners, at least one component of their detoxification systems is responsive to these compounds.