Aryloxyphenoxypropionate Herbicides Research Articles

Differential Resistance to Acetyl-CoA Carboxylase Inhibitors in Rice: Insights from Two Distinct Target-Site Mutations.

Weeds present a significant challenge to agricultural productivity, and acetyl-CoA carboxylase (ACCase)-inhibiting herbicides have proven to be effective in managing weed populations in rice fields. To develop ACCase-inhibiting herbicide-resistant rice, we generated mutants of rice ACCase (OsACC) featuring Ile-1792-Leu or Gly-2107-Ser substitutions through ethyl methyl sulfonate (EMS) mutagenesis. The Ile-1792-Leu mutant displayed cross-resistance to aryloxyphenoxypropionate (APP) and phenylpyrazoline (DEN) herbicides, whereas the Gly-2107-Ser mutants primarily exhibited cross-resistance to APP herbicides with diminished resistance to the DEN herbicide. In vitro assays of the OsACC activity revealed an increase in resistance to haloxyfop and quizalofop, ranging from 4.84- to 29-fold in the mutants compared to that in wild-type. Structural modeling revealed that both mutations likely reduce the binding affinity between OsACC and ACCase inhibitors, thereby imparting resistance. This study offers insights into two target-site mutations, contributing to the breeding of herbicide-resistant rice and presenting alternative weed management strategies in rice cultivation.

Resistance patterns and molecular basis to ACCase-inhibiting herbicides in Digitaria ciliaris var. chrysoblephara from China

Abstract Digitaria ciliaris var. chrysoblephara (Fig. & De Not.) R.R. Stewart is an annual xeromorphic weed that severely infests direct-seeded rice fields in China. Herbicide resistance is emerging in D. ciliaris var. chrysoblephara owing to extensive and recurrent use of acetyl-CoA carboxylase (ACCase)-inhibiting herbicide metamifop. In this study, a total of 53 D. ciliaris var. chrysoblephara populations randomly sampled from direct-seeded rice fields across Jiangsu Province were investigated for metamifop resistance and potential resistance-endowing mutations. Single-dose assays revealed that 17 (32.1%) populations evolved resistance to metamifop and 5 (9.4%) populations were in the process of developing resistance. Resistance index (RI) of metamifop-resistant populations ranged from 2.7 to 32.1. Amino acid substitutions (Ile-1781-Leu, Trp-2027-Cys/Ser, and Ile-2041-Asn) in ACCase genes were detected in resistant D. ciliaris var. chrysoblephara plants, and caused various cross-resistance patterns to ACCase-inhibiting herbicides. All of four resistant populations (YC07, YZ09, SQ03, and HA06), with different ACCase mutations, exhibited cross-resistance to the aryloxyphenoxypropionate (APP) herbicides cyhalofop-butyl (RIs, 10.0 to 19.9), fenoxaprop-P-ethyl (RIs, 53.7 to 132.8), and haloxyfop-P-methyl (RIs, 6.2 to 62.6), and the phenylpyrazoline (DEN) pinoxaden (RIs, 2.3 to 5.4), but responded differently to the cyclohexanedione (CHD) herbicides clethodim and sethoxydim. It is noteworthy that four post-emergence herbicides used for rice cropping, including bispyribac-sodium, pyraclonil, quinclorac, and anilofos, showed poor control effect against D. ciliaris var. chrysoblephara, suggesting few alternations for managing this weed in rice fields except ACCase inhibitors. In conclusion, this work demonstrated that the D. ciliaris var. chrysoblephara had developed resistance to ACCase-inhibiting herbicides in rice cultivation of China, and target-site amino acid substitutions in ACCase were primarily responsible for metamifop resistance.

Life cycle assessment of hepatotoxicity induced by cyhalofop-butyl in environmental concentrations on zebrafish in light of gut-liver axis

Cyhalofop-butyl (CB) poses a significant threat to aquatic organisms, but there is a discrepancy in evidence about hepatotoxicity after prolonged exposure to environmental levels. The aim of this study was to investigate long-term hepatotoxicity and its effects on the gut-liver axis through the exposure of zebrafish to environmental concentrations of CB (0.1,1,10 μg/L) throughout their life cycle. Zebrafish experienced abnormal obesity symptoms and organ index after a prolonged exposure of 120 days. The gut-liver axis was found to be damaged both morphologically and functionally through an analysis of histology, electron microscopy subcellular structure, and liver function. The disruption of the gut-liver axis inflammatory process by CB is suggested by the rise in inflammatory factors and the alteration of inflammatory genes. Furthermore, there was a noticeable alteration in the blood and gut-liver axis biochemical parameters as well as gene expression linked to lipid metabolism, which may led to an imbalance in the gut flora. In conclusion, the connection between the gut-liver axis, intestinal microbiota, and liver leads to the metabolic dysfunction of zebrafish exposed to long-term ambient concentrations of CB, and damaged immune system and liver lipid metabolism. This study gives another knowledge into the hepatotoxicity component of long haul openness to ecological centralization of CB, and might be useful to assess the potential natural and wellbeing dangers of aryloxyphenoxypropionate herbicides.

Synthesis, Herbicidal Activity, and Molecular Mode of Action Evaluation of Novel Aryloxyphenoxypropionate/Amide Derivatives Containing a Quinazolinone Moiety.

To develop aryloxyphenoxypropionate herbicides with a novel structure and improved activity, a total of 39 aryloxyphenoxypropionate/amide derivatives containing quinazolinone moiety were synthesized and further bioevaluated. The bioassay results in the greenhouse showed that most of the target compounds had good herbicidal activity under postemergence conditions, of which, QPP-I-6 displayed excellent herbicidal activity against Echinochloa crusgalli, Digitaria sanguinalis, Spartina alterniflora, Eleusine indica, and Pennisetum alopecuroides with inhibition rates >90% at a dosage of 187.5 g ha-1. More importantly, QPP-I-6 displayed higher crop safety to Gossypium hirsutum, Glycine max, and Arachis hypogaea than the commercial herbicide quizalofop-p-ethyl. Studying the molecular mode of action by phenotypic observation, membrane permeability evaluation, transcriptomic analysis, and in vivo ACCase activity evaluation reveals that QPP-I-6 is a novel ACCase inhibitor. The present work demonstrates that QPP-I-6 can serve as a lead compound for further developing novel ACCase-inhibiting herbicides.

Changes in physiology, antioxidant system, and gene expression in Microcystis aeruginosa under fenoxaprop-p-ethyl stress.

Fenoxaprop-p-ethyl (FE) is one of the typical aryloxyphenoxypropionate herbicides. FE has been widely applied in agriculture in recent years. Human health and aquatic ecosystems are threatened by the cyanobacteria blooms caused by Microcystis aeruginosa, which is one of the most common cyanobacteria responsible for freshwater blooming. Few studies have been reported on the physiological effects of FE on M. aeruginosa. This study analyzed the growth curves, the contents of chlorophyll a and protein, the oxidative stress, and the microcystin-LR (MC-LR) levels of M. aeruginosa exposed to various FE concentrations (i.e., 0, 0.5, 1, 2, and 5mg/L). FE was observed to stimulate the cell density, chlorophyll a content, and protein content of M. aeruginosa at 0.5- and 1-mg/L FE concentrations but inhibit them at 2 and 5mg/L FE concentrations. The superoxide dismutase and catalase activities were enhanced and the malondialdehyde concentration was increased by FE. The intracellular (intra-) and extracellular (extra-) MC-LR contents were also affected by FE. The expression levels of photosynthesis-related genes psbD1, psaB, and rbcL varied in response to FE exposure. Moreover, the expressions of microcystin synthase-related genes mcyA and mcyD and microcystin transportation-related gene mcyH were significantly inhibited by the treatment with 2 and 5mg/L FE concentrations. These results might be helpful in evaluating the ecotoxicity of FE and guiding the rational application of herbicides in modern agriculture.

Characterization of two novel hydrolases from Sphingopyxis sp. DBS4 for enantioselective degradation of chiral herbicide diclofop-methyl

Diclofop-methyl, an aryloxyphenoxypropionate (AOPP) herbicide, is a chiral compound with two enantiomers. Microbial detoxification and degradation of various enantiomers is garnering immense research attention. However, enantioselective catabolism of diclofop-methyl has been rarely explored, especially at the molecular level. This study cloned two novel hydrolase genes (dcmA and dcmH) in Sphingopyxis sp. DBS4, and characterized them for diclofop-methyl degradation. DcmA, a member of the amidase superfamily, exhibits 26.1–45.9% identity with functional amidases. Conversely, DcmH corresponded to the DUF3089 domain-containing protein family (a family with unknown function), sharing no significant similarity with other biochemically characterized proteins. DcmA exhibited a broad spectrum of substrates, with preferential hydrolyzation of (R)-(+)-diclofop-methyl, (R)-(+)-quizalofop-ethyl, and (R)-(+)-haloxyfop-methyl. DcmH also preferred (R)-(+)-quizalofop-ethyl and (R)-(+)-haloxyfop-methyl degradation while displaying no apparent enantioselective activity towards diclofop-methyl. Using site-directed mutagenesis and molecular docking, it was determined that Ser175 was the fundamental residue influencing DcmA's activity against the two enantiomers of diclofop-methyl. For the degradation of AOPP herbicides, DcmA is an enantioselective amidase that has never been reported in research. This study provided novel hydrolyzing enzyme resources for the remediation of diclofop-methyl in the environment and deepened the understanding of enantioselective degradation of chiral AOPP herbicides mediated by microbes.

Developmental toxicity and metabolomics analyses of zebrafish (Danio rerio) embryos exposed to Fenoxaprop-p-ethyl.

Fenoxaprop-p-ethyl (FEN) is an aryloxy phenoxy propionate herbicide that has been widely used in paddy fields. Previous studies have indicated that FEN is highly toxic to aquatic organisms, but little is known about the developmental effects of FEN. This study investigated acute and developmental toxicity, malondialdehyde (MDA) levels, superoxide dismutase (SOD) and catalase (CAT) activities, and metabolomic analyses in zebrafish embryos after 96h of exposure. FEN exhibited high acute toxicity to zebrafish embryos and larvae. Exposure to FEN could reduce heartbeat and hatching rates and increase malformation rates in embryos. Oxidative damage was also caused in embryos. The results of metabolomics analysis showed that 102 differentially abundant metabolites were found in zebrafish embryos in the 0.05mg/L FEN treatment group, and 60 differentially abundant metabolites were found in the 0.20mg/L FEN treatment group. These differentially abundant metabolites mainly belonged to 9 metabolic pathways, of which folate pathways and ABC transport protein pathways had the greatest impact. These results suggested that FEN induced high acute and developmental toxicity in zebrafish embryos.

First report of target-site resistance to ACCase-inhibiting herbicides in Bromus tectorum L.

The prevalent and repeated use of acetyl-coenzyme A carboxylase (ACCase)-inhibiting herbicides for Bromus tectorum L. control in fine fescue (Festuca L. spp) grown for seed has selected ACCase-resistant B. tectorum populations. The objectives of this study were to (1) evaluate the response of nine B. tectorum populations to the ACCase inhibitors clethodim, sethoxydim, fluazifop-P-butyl, and quizalofop-P-ethyl and the acetolactate synthase (ALS) inhibitor sulfosulfuron and (2) characterize the resistance mechanisms. Bromus tectorum populations were confirmed to be resistant to the ACCase-inhibiting herbicides tested. The levels of resistance varied among the populations for clethodim (resistance ratio, RR = 5.1-14.5), sethoxydim (RR = 18.7-44.7), fluazifop-P-butyl (RR = 3.1-40.3), and quizalofop-P-ethyl (RR = 14.5-36). Molecular investigations revealed that the mutations Ile2041Thr and Gly2096Ala were the molecular basis of resistance to the ACCase-inhibiting herbicides. The Gly2096Ala mutation resulted in cross-resistance to the aryloxyphenoxypropionate (APP) herbicides fluazifop-P-butyl and quizalofop-P-ethyl, and the cyclohexanedione (CHD) herbicides clethodim, and sethoxydim, whereas Ile2041Thr mutation resulted in resistance only to the two APP herbicides. All B. tectorum populations were susceptible to sulfosulfuron (RR = 0.3-1.7). This is the first report of target-site mutations conferring resistance to ACCase-inhibiting herbicides in B. tectorum. The results of this study suggest multiple evolutionary origins of resistance and contribute to understanding the patterns of cross-resistance to ACCase inhibitors associated with different mutations in B. tectorum. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Ile-1781-Leu Target Mutation and Non-Target-Site Mechanism Confer Resistance to Acetyl-CoA Carboxylase-Inhibiting Herbicides in Digitaria ciliaris var. chrysoblephara.

Digitaria ciliaris var. chrysoblephara is a xerophytic weed severely invading rice fields along with the application of rice mechanical direct seeding technology in China. This study identified one resistant population (M5) with an Ile-1781-Leu substitution in ACCase1 showing broad-spectrum resistance to three chemical classes of ACCase-inhibiting herbicides, including metamifop, cyhalofop-butyl, fenoxaprop-p-ethyl, haloxyfop-p-methyl, clethodim, sethoxydim, and pinoxaden. The other two populations, M2 and M4, without any resistance-responsible mutations, only exhibited resistance to aryloxyphenoxypropionate (APP) herbicides cyhalofop-butyl and fenoxaprop-p-ethyl. Pre-treatment with the cytochrome P450 monooxygenase (P450) inhibitor PBO significantly reduced the cyhalofop-butyl resistance by 43% in the M2 population. Pre-emergence weed control with soil-applied herbicides, such as pretilachlor, pendimethalin, and oxadiazon, can effectively inhibit the germination and growth of D. ciliaris var. chrysoblephara. The present study reported a xerophytic weed species invading rice fields featuring broad-spectrum resistance to ACCase-inhibiting herbicides as a result of Ile-1781-Leu mutation of ACCase. Both target- and P450-involved non-target-site mechanisms may be contributing to resistance in D. ciliaris var. chrysoblephara species.

Mechanism of metamifop resistance in Digitaria ciliaris var. chrysoblephara from Jiangsu, China.

Digitaria ciliaris var. chrysoblephara is one of the most competitive and problematic grass weeds in China. Metamifop is an aryloxyphenoxypropionate (APP) herbicide that inhibits the activity of acetyl-CoA carboxylase (ACCase) of sensitive weeds. Following the introduction of metamifop to China in 2010, it has been continuously used in rice paddy fields, thereby substantially increasing selective pressure for resistant D. ciliaris var. chrysoblephara variants. Here, populations of D. ciliaris var. chrysoblephara (JYX-8, JTX-98, and JTX-99) were observed to be highly resistant to metamifop, with resistance index (RI) values of 30.64, 14.38, and 23.19, respectively. Comparison of resistant and sensitive population ACCase gene sequences revealed that a single nucleotide substitution from TGG to TGC resulted in an amino acid substitution from tryptophan to cysteine at position 2,027 in the JYX-8 population. No corresponding substitution was observed for JTX-98 and JTX-99 populations. The ACCase cDNA of D. ciliaris var. chrysoblephara was successfully obtained by PCR and RACE methods, representing the first amplification of full length ACCase cDNA from Digitaria spp. Investigation of the relative expressions of ACCase gene revealed the lack of significant differences between sensitive and resistant populations before and after herbicide treatments. ACCase activities in resistant populations were less inhibited than in sensitive populations and recovered to the same or even higher levels compared to untreated plants. Whole-plant bioassays were also conducted to assess resistance to other ACCase inhibitors, acetolactate synthase (ALS) inhibitors, auxin mimic herbicide, and protoporphyrinogen oxidase (PPO) inhibitor. Cross-resistance and some multi-resistance were observed in the metamifop-resistant populations. This study is the first to focus on the herbicide resistance of D. ciliaris var. chrysoblephara. These results provide evidence for a target-site resistance mechanism in metamifop-resistant D. ciliaris var. chrysoblephara, while providing a better understanding of cross- and multi-resistance characteristics of resistant populations that will help in the management of herbicide-resistant D. ciliaris var. chrysoblephara.

Identifying a Detoxifying Uridine Diphosphate Glucosyltransferase (UGT), MdUGT83K2, Which Can Glycosylate the Aryloxyphenoxypropionate Herbicide

Glycosylation is a common modification reaction in plants. The products obtained upon glycosylation have different biological functions, making glycosylation an important mechanism affecting and regulating the balance of plant growth and metabolism. In this study, we first speculated that Group I in the apple glycosyltransferase family may have a predicted function like UGT83A1, according to gene chip data published online. Subsequently, by real-time PCR (polymerase chain reaction), we analyzed whether the expression of nine glycosyltransferase genes in Group I was induced by our previously reported ACCase (Acetyl-CoA carboxylase) inhibition-based herbicide QPP ((R)-ethyl·2-(4-((6-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)oxy) phenoxy) propanoate). It was found that expression of the MdUGT83K2 gene in Group I was significantly increased by QPP. In order to determine whether MdUGT83K2 can glycosylate QPP, we confirmed the enzymatic reaction of MdUGT83K2 in vitro and the presence of QPP glycosides in MdUGT83K2 transgenic apple seedlings by HPLC (High Performance Liquid Chromatography), and found that MdUGT83K2 can transfer glucose to QPP in vivo, which is glycosylated. In this work, we identified a novel apple glycosyltransferase, MdUGT83K2, which functions to glycosylate the ACCase-inhibiting herbicide QPP and may be involved in plant detoxification. Key Contribution: A novel apple glycosyltransferase, MdUGT83K2, was identified, which may be involved in plant detoxification by glycosylation modification of the ACCase-inhibiting herbicide.

Facile synthesis of dual-hydrolase encapsulated magnetic ZIF-8 composite for efficient removal of multi-pesticides induced pollution in water

Multi-pesticides pollution induced by organophosphorus insecticides (OPs) and aryloxyphenoxypropionate herbicides (AOPPs) has become a significant challenge in bioremediation of water pollution due to their prolonged and over application. Though a number of physical, chemical, and biological approaches have been developed for different pesticides, the explorations usually focus on eliminating single pesticide pollution. Herein, a heterostructure nanocomposite OPH/QpeH@mZIF-8, encapsulating OPs hydrolase OPH and AOPPs hydrolase QpeH in the magnetic zeolitic imidazolate frameworks-8 (mZIF-8), was synthesized through a facile one-pot method in aqueous solution. The immobilized OPH and QpeH in mZIF-8 showed high activities towards the two most common OPs and AOPPs, i.e., chlorpyrifos and quizalofop-P-ethyl, which were hydrolyzed to 3,5,6-Trichloro-2-pyridino (TCP) and quizalofop acid, respectively. Moreover, the magnetic nanocatalyst possessed great tolerance towards broad pH range, high temperatures, and different chemical solvents and excellent recyclability. More importantly, compared to free OPH and QpeH, OPH/QpeH@mZIF-8, with significantly enhanced degradation capability, exhibited enormous potential for simultaneous removal of chlorpyrifos and quizalofop-p-ethyl from the surface and industrial wastewater. Overall, the study demonstrates the applicability of this strategy for utilizing magnetic nanocatalysts encapsulating multiple enzymes due to its simplicity, high efficiency, and economic benefits to removing pesticide compound pollution from various water resources.

Effects of haloxyfop-p-methyl on the developmental toxicity, neurotoxicity, and immunotoxicity in zebrafish

Pesticides are extensively used in agricultural production, and their residues in soil, water, and agricultural products have become a threat to aquatic ecosystem. In this study, the toxicity of haloxyfop-p-methyl, an aryloxyphenoxypropionate herbicide was studied using the model animal zebrafish. The development of zebrafish larvae was affected by haloxyfop-p-methyl including spinal deformities, decreased body length, slow heart rate, and large yolk sac area. Behavior analysis revealed that behavior activity of larvae was weakened significantly including shortened displacement distance, reduced swimming speed, increased angular speed winding degrees, in accordance with higher AChE activity. Besides, exposure to haloxyfop-p-methyl could induce oxidative stress companied by the increased intents of ROS, MDA and increased activities of CAT and SOD. In immunotoxicity, haloxyfop-p-methyl not only reduced the innate immune cells such as neutrophils and macrophages, but also affected T cells mature in thymus. Furthermore, haloxyfop-p-methyl could induce neutrophils apoptosis, accompanied with the upregulation of the expression of proapoptotic protein such as Bax and P53 and the downregulation of the expression of antiapoptotic protein Bcl-2. In addition, haloxyfop-p-methyl could induce the expression of Jak, STAT and proinflammatory cytokine genes (IFN-γ, TNF-α, and IL-8). These results indicate that haloxyfop-p-methyl induces developmental toxicity, neurotoxicity, and immunotoxicity in zebrafish, providing a perspective on the toxicological mechanism of haloxyfop-p-methyl in teleosts.

Weed Control, Rice Safety, and Mechanism of the Novel Paddy Field Herbicide Glyamifop

Glyamifop (R&D code: FG001), (R)-(2-(4-(6-chlorobenzoxazol-2-oxy) phenoxy) propionyl) glycine ethyl ester is a newly developed aryloxyphenoxypropionate (HRAC Group 1) herbicide for weed control in paddy fields. This work determined the effect of Glyamifop on weeds and its safety for rice in the glasshouse. Glyamifop controlled the common gramineous weeds in paddy fields at 100 g a.i. ha−1: the fresh weight inhibition rates of Echinochloa crus-galli, Leptochloa chinensis, Setaria viridis, Eragrostis japonica, Digitaria sanguinalis and Panicum bisulcatum were all above 90%. It has almost no inhibitory effect on broad-leaved and cyperaceae weeds, such as Eclipta prostrata and Cyperus iria. Glyamifop inhibited cyhalofop-butyl-resistant L. chinensis, penoxsulam-resistant E. crus-galli and quinclorac-resistant E. crusgalli var. zelayensis by 100%, 99.98% and 96.37%, respectively, at 100 g a.i. ha−1, based on the fresh weight. The selectivity index of Glyamifop foliage treatment in the rice varieties japonica ‘Huaidao 5’, indica ‘Xiangliangyou 900’ and glutinous ‘Zhennuo 29’ was 5.93, 6.81 and 4.91, respectively; therefore, Glyamifop is safe for the 3 different rice varieties. Fresh weight rice inhibition rates were 7.18%, 2.99% and 7.93% at the 2.5-, 3.5- and 5.5-leaf stage, respectively, and the selectivity index was 5.18, 6.04 and 7.93, respectively, indicating that Glyamifop was safe for rice at these leaf stages. L. chinensis ACCase activity decreased with increasing Glyamifop concentration, and the inhibitory effect was similar to that of cyhalofop acid; this confirmed that Glyamifop is an ACCase inhibitor. In conclusion, Glyamifop has potential for the management of gramineous weeds as it has good activity against weeds that are resistant to common herbicides in paddy fields.

Metabolic Resistance to Acetyl-CoA Carboxylase-Inhibiting Herbicide Cyhalofop-Butyl in a Chinese Echinochloa crus-galli Population

A population of Echinochloa crus-galli (L.) P. Beauv obtained from direct-seeding rice fields in Jiangxi Province, China, exhibited high resistance levels (13.5-fold) to the acetyl-CoA carboxylase (ACCase)-inhibiting herbicide cyhalofop-butyl. Compared with the susceptible (S) population, this resistant (R) population evolved a cross-resistance to aryloxyphenoxypropionates (APPs) herbicides metamifop (2.9-fold) and fenoxapro-p-ethyl (4.1-fold), cyclohexanediones (CHDs) herbicide clethodim (4.7-fold), phenyl pyrazoline (DEN) herbicide pinoxaden (6.4-fold), and evolved multiple-resistance to acetolactate synthase (ALS)-inhibiting herbicide penoxsulam (3.6-fold), and auxin mimic herbicides quinclorac (>34.7-fold) and florpyrauxifen-benzyl (2.4-fold). ACCase gene sequencing did not reveal the existence of any known mutation point conferring with herbicide resistance. In addition, three metabolic inhibitors—one glutathione—S-transferase (GST) inhibitor (NBD-Cl), and two cytochrome P450 inhibitors (malathion and PBO)—did not reverse the cyhalofop-butyl resistance. Furthermore, enhanced metabolic rates of more than 60% 24 h after treatment with the active compound cyhalofop acid was observed in R plants compared to S plants. Hence, enhanced metabolism activity endows a non-target-site resistance to cyhalofop-butyl in the R population of E. crus-galli. Future research will be required to determine what metabolizing enzyme genes are responsible for cyhalofop-butyl resistance in E. crus-galli.

Transcriptome analysis reveals hepatotoxicity in zebrafish induced by cyhalofop‑butyl

Cyhalofop‑butyl is a highly effective aryloxyphenoxypropionate herbicide and widely used for weed control in paddy fields. With the increasing residue of cyhalofop‑butyl, it poses a threat to the survival of aquatic organisms. Here, we investigated the effect of cyhalofop‑butyl on zebrafish to explore its potential hepatotoxic mechanism. The results showed that cyhalofop‑butyl induced hepatocyte degeneration, vacuolation and necrosis of larvae after embryonic exposure for 4 days and caused liver atrophy after 5 days. Meanwhile, the activities of enzymes related to liver function were significantly increased by 0.2 mg/L cyhalofop‑butyl and higher, such as alanine transaminase (ALT) and aspartate transaminase (AST). And the contents of triglyceride (TG) involved in lipid metabolism were significantly decreased by 0.4 mg/L cyhalofop-buty. The expression of genes related to liver development was also significantly down-regulated. Furthermore, transcriptome results showed that the pathways involved in metabolism, immune system and endocrine system were significantly impacted, which may be related to hepatoxicity. To sum up, the present study demonstrated the hepatoxicity caused by cyhalofop-buty and its underlying mechanism. The results may provide new insights for the risk of cyhalofop‑butyl to aquatic organisms and new horizons for the pathogenesis of hepatotoxicity.

Cross-resistance of barnyardgrass [Echinochloa crus-galli (L.) P. Beauv.] to aryloxyphenoxypropionate herbicides

Managing emerged weeds that have evolved resistance to acetyl CoA carboxylase (ACCase)-inhibiting herbicides is a challenging task. A dose-response experiment was conducted on barnyardgrass biotypes resistant (R) and susceptible (S) to three aryloxyphenoxypropionate herbicides cyhalofop-butyl (CyB), fenoxaprop-ethyl (FeE), and quizalofop-ethyl (QuE) along with investigations into the potential resistance mechanism of these biotypes. The tested R barnyardgrass biotypes had strong resistance to CyB and FeE (resistant/susceptible ratio: 7.9–14.4) but weak resistance to QuE (resistant/susceptible ratio: 2.4–3.1). Absorption, translocation, and total metabolism of CyB and QuE were not associated with differences among S and R barnyardgrass biotypes. However, differences between S and R barnyardgrass were observed in production of active acid forms of each herbicide (cyhalofop-acid and quizalofop-acid). Production of cyhalofop-acid was >1.6-fold less in R barnyardgrass (3–8%) for 24 h after herbicide application than in the S barnyardgrass (8–16%). Meanwhile, production of quizalofop-acid was less in R barnyardgrass (< 14%) throughout the study period than in the S barnyardgrass (< 22%). Sequencing results of ACCase gene showed no difference between S and R barnyardgrass. Overall results show that a non-target-site resistance mechanism altering metabolism of CyB and QuE likely contributes to resistance of the barnyardgrass biotypes to these herbicides.

TREATMENT WITH FLUAZIFOP-P-BUTYL HERBICIDE INDUCES OXIDATIVE STRESS IN PHASEOLUS VULGARIS L.

Fluazifop-p-butyl is an aryloxyphenoxy-propionate herbicide used for broad-spectrum weed control in Phaseolus vulgaris and other legume crops. In this study, field experiment was conducted to evaluate some physiological responses of Phaseolus vulgaris to herbicide treatment. Parameters measured includes: whole plant biomass, chlorophyll content, lipid peroxidation, relative water content of the leaves, carbohydrate content of the leaves, and antioxidant enzymes activities. The results showed that fluazifop-P-butyl significantly decreased the plant biomass, relative water content of the leaves, total chlorophyll and carbohydrate contents of the treated plant compared with the control. In contrast, there was a significance increase in malondialdehyde level as well as activities of antioxidant enzymes. The results of this work showed that herbicides caused physiological disorders in non-target plants by inducing oxidative stress that may lead to poor growth and yield.

Magnetic ZIF-8-Based Mimic Multi-enzyme System as a Colorimetric Biosensor for Detection of Aryloxyphenoxypropionate Herbicides.

In the present study, a magnetic mimic multi-enzyme system was developed by encapsulating the aryloxyphenoxypropionate (AOPP) herbicide hydrolase QpeH and alcohol oxidase (AOx) in zeolitic imidazolate framework (ZIF-8) nanocrystals with magnetic Fe3O4 nanoparticles (MNPs) to detect AOPP herbicides. The structural, protein loading capacity and loading ratio, porosity, and magnetic properties of QpeH/AOx@mZIF-8 were characterized by scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analysis, nitrogen sorption, and vibrating sample magnetometry. An AOPP herbicide colorimetric biosensor made with QpeH/AOx@mZIF-8 had the highest sensitivity toward quizalofop-P-ethyl (QpE) with a limit of detection of 8.2 μM. This system was suitable to detect two other AOPP herbicides, including fenoxaprop-P-ethyl (FpE) and haloxyfop-P-methyl (HpE). The practical application of the biosensor was verified through quantitative analysis of QpE residues in industrial wastewater and field soils. Furthermore, QpeH/AOx@mZIF-8 exhibited excellent long-term storage stability (at least 50 days), easy separation by magnet, and reusability (at least 10 cycles), supporting its promising role in simple and low-cost detection of AOPP herbicides in real environmental samples.

Detecting the effect of ACCase-targeting herbicides on ACCase activity utilizing a malachite green colorimetric functional assay

AbstractResearch was conducted using a functional malachite green colorimetric assay to evaluate acetyl-coenzyme A carboxylase (ACCase) activity previously identified as resistant to sethoxydim and select aryloxyphenoxypropionate (FOPs) herbicides, fenoxaprop, and fluazifop. Two resistant southern crabgrass [Digitaria ciliaris (Retz.) Koeler] biotypes, R1 and R2, containing an Ile-1781-Leu amino acid substitution and previously identified as resistant to sethoxydim, pinoxaden, and fluazifop but not clethodim was utilized as the resistant chloroplastic ACCase source compared with known susceptible (S) ACCase. Dose-response studies with sethoxydim, clethodim, fluazifop-p-butyl, and pinoxaden (0.6 to 40 µM) were conducted to compare the ACCase–herbicide interactions of R1, R2, and S using the malachite green functional assay. Assay results indicated that R biotypes required more ACCase-targeting herbicides to inhibit ACCase activity compared with S. IC50 values of all four herbicides for R biotypes were consistently an order of magnitude greater than those of S. No sequencing differences in the carboxyltransferase domain was observed for R1 and R2; however, R2 IC50 values were greater across all herbicides. These results indicate the malachite green functional assay is effective in evaluating ACCase activity of R and S biotypes in the presence of ACCase-targeting herbicides, which can be used as a replacement for the 14C-based radiometric functional assays.