Acetolactate Synthase Research Articles

Effect of herbicide programs on control and seed production of multiple herbicide–resistant Palmer amaranth (Amaranthus palmeri) in corn resistant to 2,4-D/glufosinate/glyphosate

Abstract Multiple herbicide–resistant (MHR) Palmer amaranth is among the most problematic summer annual broadleaf weeds in Nebraska and several other states. A new MHR corn cultivar (resistant to 2,4-D/glufosinate/glyphosate, also known as Enlist corn) has been commercially available in the United States since 2018. Growers are searching for herbicide programs for control and reduce seed production of MHR Palmer amaranth among Enlist corn crops. The objectives of this study were to evaluate herbicides applied preemergence, early postemergence, or preemergence followed by (fb) late postemergence for the management of MHR Palmer amaranth in Enlist corn fields and to assess their effect on Palmer amaranth biomass, density, seed production, and corn yield. Field experiments were conducted near Carleton, NE, in 2020 and 2021, in a grower’s field of Enlist corn infested with acetolactate synthase–inhibitor/atrazine/glyphosate–resistant Palmer amaranth. Herbicides applied preemergence, such as flufenacet/isoxaflutole/thiencarbazone-methyl, acetochlor/clopyralid/flumetsulam, or acetochlor/clopyralid/mesotrione, provided 75% to 99% control of Palmer amaranth 30 d after preemergence. Preemergence fb late postemergence herbicides resulted in 94% Palmer amaranth control 90 d after late postemergence, reduced weed density to 0 to 8 plants m−2 30 d after late postemergence, and reduced biomass to 2 to 14 g m−2 15 d after late postemergence compared to preemergence-only (59% control, 0 to 15 plants m−2, and 4 to 123 g m−2) and early postemergence–only herbicides (78% control, 6 to 30 plants m−2, and 8 to 25 g m−2). Based on contrast analysis, Palmer amaranth seed production was reduced to 14,050 seeds m–2 in preemergence fb late postemergence herbicide programs compared with 325,490 seed m–2 in preemergence-only and 376,750 seed m–2 in early postemergence–only programs. Based on orthogonal contrast, higher corn yield of 12,340 and 11,730 kg ha−1 was obtained with preemergence fb late postemergence herbicide programs compared with preemergence-only (10,840 and 11,510 kg ha−1) and early postemergence–only programs (10,850 and 10,030 kg ha−1) in 2020 and 2021, respectively.

Biosynthesis of Diverse Ephedra-Type Alkaloids via a Newly Identified Enzymatic Cascade.

Ephedra-type alkaloids represent a large class of natural and synthetic phenylpropanolamine molecules with great pharmaceutical values. However, the existing methods typically rely on chemical approaches to diversify the N-group modification of Ephedra-type alkaloids. Herein, we report a 2-step enzymatic assembly line for creating structurally diverse Ephedra-type alkaloids to replace the conventional chemical modification steps. We first identified a new carboligase from Bacillus subtilis (BsAlsS, acetolactate synthase) as a robust catalyst to yield different phenylacetylcarbinol (PAC) analogs from diverse aromatic aldehydes with near 100% conversions. Subsequently, we screened imine reductases (IREDs) for the reductive amination of PAC analogs. It was found that IRG02 from Streptomyces albidoflavus had good activities with conversions ranging from 37% to 84% for the reductive alkylamination with diverse amine partners such as allylamine, propargylamine, and cyclopropylamine. Overall, 3 new bio-modifications at the N-group of Ephedra-type alkaloids were established. Taken together, our work lays a foundation for the future implementation of biocatalysis for synthesizing structurally diverse Ephedra-type alkaloids with potential new pharmaceutical applications.

Management of Herbicide resistant Phalaris minor: A Review

Littleseed canarygrass (Phalaris minor Retz), an annual species of the family Poaceae, is a major weed problem in many wheat-producing countries. The excessive dependence on herbicides for its control led to the evolution of multiple herbicides resistance and yield losses. The first case of resistant P. minor in India was to the Photosystem-II inhibitor isoproturon in the early 1990’s. P. minor has been found resistant to Acetolactate Synthase (ALS) inhibitors, Acetyl-CoA Carboxylase (ACCase) inhibitors, Photosystem II inhibitors. It is highly competitive and in severe cases can cause complete crop failure, forcing farmers to harvest their crop as fodder or ploughing the standing crop along with weeds. For management of these multiple herbicide resistant P. minor, pyroxasulfone, pendimethalin, flumioxazin, trifluralin and flufenacet can be used as alternative herbicides. Moreover, in-depth biological and ecological studies on P. minor should be carried out for devising integrated weed management strategies. The long-term strategies of efficient weed management and sustainability of wheat production should include the use of alternative herbicides, herbicide mixture, crop rotation along with other non-chemical methods like manual removal of weeds, mulching, Summer ploughing etc providing the competitive advantages to the wheat crop over P. minor, at the same time increasing the profitability of the farmer by controlling the weed effectively and reducing the selection pressure on the weeds which will inturn reduce the resistance development in weeds

Mutação Try574 conferindo resistência cruzada a herbicidas inibidores da ALS em nabiça

ABSTRACT Understanding how weeds resist herbicides, their resistance mechanisms, and alternative control methods are crucial for managing herbicide-resistant weeds. This study aims to unravel the resistance mechanism of a Raphanus raphanistrum biotype to acetolactate synthase (ALS) inhibitors. To this end, dose-response studies, DNA sequencing, and metabolic pathway verification were conducted. ALS-inhibiting herbicides showed low efficacy in controlling this biotype, confirming cross-resistance. Sequencing of the ALS enzyme revealed the presence of the previously reported Try-574-Leu mutation, known to confer cross-resistance to this mode of action. However, the metabolization verification assay demonstrated that this mechanism did not contribute to the observed resistance. Chemical control studies with alternative herbicides yielded promising results, indicating the potential for effective management of the resistant biotype. Our findings showed that the wild radish biotype used exhibits cross-resistance to ALS-inhibiting herbicides due to the presence of the Try-574-Leu mutation in the target enzyme. Notably, herbicides with alternative mechanisms of action prove highly effective in controlling this resistant biotype, offering valuable options for weed management strategies.

Synthesis, Herbicidal Activity against Barnyard Grass, and Photolytic Behavior of Aryl 2,6-Dipyrimidinoxybenzoates.

In this study, we investigated the characteristics and herbicidal potential of bispyribac phenolic esters, which belong to the 2-(pyrimidin-2-yloxy)benzoic acid (PYB) class of acetohydroxyacid synthase (AHAS-)-inhibiting herbicides. These herbicides are primarily used for controlling Poaceae and broadleaf weeds. Among them, bispyribac-sodium stands out as a representative in this class. Surprisingly, other bispyribac esters, including alkanol and phenol esters exhibit considerably reduced herbicidal activity compared to bispyribac-sodium. In contrast, oxime esters (e.g., pyribenzoxim) demonstrate high activity. To further understand and develop novel PYB herbicides, we synthesized and screened a series of bispyribac phenolic esters while investigating their photochemical behaviors. Several compounds displayed excellent herbicidal activity, with compounds Ia-19 and Ic showing impressive 90% effective dosages for fresh weight inhibition of barnyard grass, measuring 0.55 and 0.60 g a.i./hm2, respectively. These values were approximately half of bispyribac-sodium or pyribenzoxim. The results indicate that the herbicidal activity of phenolic esters is influenced by both their binding ability to the AHAS enzyme and their decomposition into bispyribac acid. For instance, bispyribac phenol ester exhibited considerably reduced receptor affinity compared to bispyribac-sodium, and faced challenges in transforming into bispyribac acid, explaining its diminished herbicidal activity. However, introducing a photosensitive nitro group led to a complete transformation. This modification improved its affinity with AHAS and accelerated its decomposition into bispyribac acid, further accelerated by photocatalysis. Consequently, nitro-containing compounds displayed heightened herbicidal activity. The findings from this study open possibilities for structural optimization of phenolic esters through quantitative structure-activity relationship analysis, potentially regulating their activity-releasing period. Furthermore, the high activity of aromatic heterocyclic esters offers new insights into developing novel PYB herbicides.

Study on the toxicity prediction model ofacetolactate synthase inhibitor herbicides based on human serum albumin and superoxide dismutase binding information

Toxicity significantly influences the successful development of drugs. Based on the toxicity prediction method (carrier protein binding information-toxicity relationship) previously established by the our group, this paper introduces information on the interaction between pesticides and environmental markers (SOD) into the model for the first time, so that the toxicity prediction model can not only predict the toxicity of pesticides to humans and animals, but also predict the toxicity of pesticides to the environment. Firstly, the interaction of acetolactate synthase inhibitor herbicides (ALS inhibitor herbicides) with human serum albumin (HSA) and superoxide dismutase (SOD) was investigated systematically from theory combined with experiments by spectroscopy methods and molecular docking, and important fluorescence parameters were obtained. Then, the fluorescence parameters, pesticides acute toxicity LD50 and structural splitting information were used to construct predictive modeling of ALS inhibitor herbicides based on the carrier protein binding information (R2 = 0.977) and the predictive modeling of drug acute toxicity based on carrier protein binding information and conformational relationship (R2 = 0.991), which had effectively predicted pesticides toxicity in humans and animals. To predict potential environmental toxicity, the predictive modeling of drug acute toxicity based on superoxide dismutase binding information was established (R2 = 0.883) by ALS inhibitor herbicides-SOD binding information, which has a good predictive ability in the potential toxicity of pesticides to the environment. This study lays the foundation for developing low toxicity pesticides.

Molecular Targets of Herbicides and Fungicides─Are There Useful Overlaps for Fungicide Discovery?

New fungicide modes of action are needed for fungicide resistance management strategies. Several commercial herbicide targets found in fungi that are not utilized by commercial fungicides are discussed as possible fungicide molecular targets. These are acetyl CoA carboxylase, acetolactate synthase, 5-enolpyruvylshikimate-3-phosphate synthase, glutamine synthase, phytoene desaturase, protoporphyrinogen oxidase, long-chain fatty acid synthase, dihydropteroate synthase, hydroxyphenyl pyruvate dioxygenase, and Ser/Thr protein phosphatase. Some of the inhibitors of these herbicide targets appear to be either good fungicides or good leads for new fungicides. For example, some acetolactate synthase and dihydropteroate inhibitors are excellent fungicides. There is evidence that some herbicides have indirect benefits to certain crops due to their effects on fungal crop pathogens. Using a pesticide with both herbicide and fungicide activities based on the same molecular target could reduce the total amount of pesticide used. The limitations of such a product are discussed.

New library of pyrimido[5,4-e][1,2,4]triazolo[1,5-c]pyrimidine derivatives: Synthesis, herbicidal activity, and molecular docking study

The development of nitrogen-containing heterocyclic compounds with novel structure and remarkable biological activity is always in demand. Herein, new library of pyrimido[5,4-e][1,2,4]triazolo[1,5-c]pyrimidine derivatives (1–98) was designed, synthesized, and characterized by Fourier Transform Infrared Spectoscopy (FT-IR), nuclear magnetic resonance (1H NMR), carbon-13 nuclear magnetic resonance (13C NMR), high-resolution mass spectroscopy (HRMS), and X-ray single-crystal diffraction. Their herbicidal activities were screened for the first time by Petri dish assay and pot experiment in a glasshouse. The results indicated that some of title compounds (e.g.,1, 4, 5, and 7–11) exhibited 100 % inhibition rate against the roots and stalks of Cucumis sativus, Raphanus sativus, Sorghum bicolor and Echinochloa crusgalli at a dosage of 100 mg/L, higher than the positive control Flumetsulam, a well-known commercial acetolactate synthase (ALS) inhibiting herbicide. 2-((2,4-Dichlorophenoxy)methyl)-10-(methylthio)-8-(pentylthio)-5-propyl-5,6-dihydropyri- mido[5,4-e][1,2,4]triazolo[1,5-c]pyrimidine 7 showed notable postemergence herbicidal activity against Abutilon theophrasti, Amaranthus spinosus, Chenopodium album, Digitaria sanguinalis, Echinochloa crusgalli, and Setaria viridis at a rate of 375 g/ha, and its activities were similar to the positive control Flumetsulam. Enzymatic bioassays revealed that 7, 8, and 40 displayed good inhibitory potency against ALS with 39.6 %, 39.1 %, and 40.9 % inhibition rate, respectively, comparable to that of Flumetsulam (43.5 % inhibition rate). Moreover, molecular docking analysis was conducted by using ALS protein and target ligand molecule 7. The results of molecular docking were consistent with the bioassay experiments, showing compound 7 with a new structure and excellent herbicidal potency was potent inhibitors of ALS.

Post-Emergence Water-Dispersal Application Provides Equal Herbicidal Activity against Echinochloa crus-galli and Rice Safety as Foliar Spraying of Penoxsulam

Penoxsulam is an acetolactate synthase (ALS)-inhibiting herbicide usually applied by post-emergence foliar spraying (PFS) for the control of Echinochloa crus-galli and numerous annual weeds in paddy fields. Herbicides applied by foliar spraying can have negative impacts on the environment, ecosystems, and human health. In this study, the response of E. crus-galli and rice to the PFS and post-emergence water-dispersal (PWD) applications of penoxsulam, and the differences in the detoxification displayed by them between the two treatment methods were compared. The results showed that the PWD application of penoxsulam provides a similar control efficacy against E. crus-galli as PFS at the 1-, 3-, and 5-leaf stages. Meanwhile, the PWD application had a higher safety for the rice. After being treated with 30 g a.i. ha−1 penoxsulam, residues were not detected in the rice treated by the PWD application method, whereas, with the PFS treatment, there was 59.0 µg/kg penoxsulam remaining. With the PFS application, there were many more residues of penoxsulam in the E. crus-galli than with the PWD method; the amount of residues was 32-fold higher 12 h after treatment. The in vitro enzyme activity assays revealed that the activities of ALS, glutathione-S-transferase (GST), and cytochrome P450 monooxygenases (P450) were increased in the PWD treatments, and were 1.5-, 1.3-, and 2.3-fold higher than with PFS 72 h after treatment. The real-time quantitative PCR (qRT-PCR) revealed that the GST1 and P450 genes, CYP81A14, CYP81A12, CYP81A18, and CYP81A21 were upregulated with the PWD application versus PFS in the E. crus-galli. In summary, these results demonstrate that the herbicidal activity was not affected by the upregulation of target and metabolic enzyme activities with the PWD application of penoxsulam. This research could contribute to application strategies reducing the risk of rice injury and environmental impacts by using water-dispersal formulations of penoxsulam.

Snapshot of the Probiotic Potential of Kluveromyces marxianus DMKU-1042 Using a Comparative Probiogenomics Approach.

Kluyveromyces marxianus is a rapidly growing thermotolerant yeast that secretes a variety of lytic enzymes, utilizes different sugars, and produces ethanol. The probiotic potential of this yeast has not been well explored. To evaluate its probiotic potential, the yeast strain Kluyveromyces marxianus DMKU3-1042 was analyzed using next-generation sequencing technology. Analysis of the genomes showed that the yeast isolates had a GC content of 40.10-40.59%. The isolates had many genes related to glycerol and mannose metabolism, as well as genes for acetoin and butanediol metabolism, acetolactate synthase subunits, and lactic acid fermentation. The strain isolates were also found to possess genes for the synthesis of different vitamins and Coenzyme A. Genes related to heat and hyperosmotic shock tolerance, as well as protection against reactive oxygen species were also found. Additionally, the isolates contained genes for the synthesis of lysine, threonine, methionine, and cysteine, as well as genes with anticoagulation and anti-inflammatory properties. Based on our analysis, we concluded that the strain DMKU3-1042 possesses probiotic properties that make it suitable for use in food and feed supplementation.

Smooth pigweed (Amaranthus hybridus) and unresolved Amaranthus spp. from Brazil resistant to glyphosate exhibit the EPSPS TAP-IVS substitution

AbstractThe presence of glyphosate-resistant smooth pigweed (Amaranthus hybridus L.) biotypes has increased in southern Brazil in recent years, presenting the triple amino acid substitution TAP-IVS in 5-enol-pyruvylshikimate-3-phosphate synthase (EPSPS) previously found in Argentina. Some of these biotypes have morphological characteristics of A. hybridus and redroot amaranth (Amaranthus retroflexus L.). The present study aimed to identify, through molecular markers, the herbicide-resistant species of Amaranthus from Brazil that have the TAP-IVS substitution and to analyze the occurrence of pollen-mediated gene flow (PMGF) as the source of the TAP-IVS substitution in these biotypes. Six biotypes were evaluated using internal transcribed spacer (ITS) sequences, of which two (AMACHY-S and CAMAQ-R) were molecularly classified as A. hybridus and four (AMACRET-S, AMACVI-S, ARRGR-R, and SAOJER-R) were unclassified. Interestingly, all the glyphosate-resistant biotypes (ARRGR-R, SAOJER-R, and CAMAQ-R) had the TAP-IVS substitution and an increase in EPSPS relative copy number; however, only CAMAQ-R was confirmed as A. hybridus. Although the biotypes ARRGR-R and SAOJER-R are closely related to A. hybridus and green pigweed (Amaranthus powellii S. Watson), their species identity could not be resolved. The biotype SAOJER-R also was resistant to acetolactate synthase (ALS)-inhibiting herbicides due to a tryptophan to leucine substitution at position 574 in ALS. The evaluation of 119,746 seedlings in an intraspecific hybridization study of A. hybridus indicated an outcrossing frequency of 0.09%. In contrast, an absence of interspecific hybridization (A. hybridus × unclassified biotype, AMACVI-S) was found after screening 111,429 offspring. Unclassified biotypes might be derived from one or more ancient hybridization events and subsequently evolved the triple mutation independently. Alternatively, such biotypes could have evolved from recent hybridization events, which occur at a frequency below the level of detection in our study.

A systematic review of chemical weed management in peanut (Arachis hypogaea) in the United States: challenges and opportunities

AbstractHerbicides are the primary tool for controlling weeds in peanut (Arachis hypogaea L.) and are crucial to sustainable peanut production in the United States. The literature on chemical weed management in peanut in the past 53 yr (1970 to 2022) in the United States was systematically reviewed to highlight the strengths and weaknesses of different herbicides and identify current research gaps in chemical weed management. Residual weed control in peanut is achieved mainly with dimethenamid-P, ethalfluralin, pendimethalin, and S-metolachlor. More recently, the use of the protoporphyrinogen oxidase inhibitor flumioxazin and acetolactate synthase inhibitors, such as diclosulam, for residual weed control in peanut has increased considerably. Postemergence broadleaf weed control in peanut is achieved mainly with acifluorfen, bentazon, diclosulam, imazapic, lactofen, paraquat, and 2,4-DB, while the graminicides clethodim and sethoxydim are the major postemergence grass weed control herbicides in peanut. Although several herbicides are available for weed control in peanut, no single herbicide can provide season-long weed control due to limited application timing, lack of extended residual activity, variability in weed control spectrum, and rotational restrictions. Therefore, effective weed management in peanut often requires herbicide mixtures and/or sequential application of preplant-incorporated, preemergence, and/or postemergence herbicides. However, the available literature showed a substantive range in herbicide efficacy due to variations in environmental conditions and flushes of weed germination across years and locations. Despite the relatively high efficacy of herbicides, the selection of herbicide-resistant weeds is another area of increasing concern. Future research should focus on developing new strategies for preventing or delaying the development of resistance and improving herbicide efficacy within the context of climate change and emerging constraints such as water shortages, rising temperatures, and increasing CO2 concentration.

ALS gene overexpression and enhanced metabolism conferring Digitaria sanguinalis resistance to nicosulfuron in China.

Crabgrass (Digitaria sanguinalis) is a common malignant weed in corn fields in China. Recently, the acetolactate synthase (ALS) inhibitor, nicosulfuron, has shown decreasing efficacy against crabgrass. In order to elucidate the molecular basis of resistance to nicosulfuron in crabgrass, we conducted bioassays, combined with gene sequence analysis, relative expression and relative copy number analysis, to characterize resistance in crabgrass populations collected from Beijing, Heilongjiang, Jilin and Anhui provinces. Whole-plant dose-response results indicated that only population collected in Heilongjiang province (HLJ) had developed low level of resistance to nicosulfuron compared with the sensitive population (SD22). No known resistant mutation of ALS gene was found in HLJ population. The real-time fluorescence quantitative PCR results showed that the ALS gene copy number did not differ significantly between the HLJ and SD22 populations. However, the ALS gene expression in the HLJ was 2.07-fold higher than that of the SD22 population at 24 h after treatment with nicosulfuron. Pretreatment with the cytochrome P450 (CYP450) inhibitor malathion, piperonyl butoxide (PBO), and the glutathione S-transferase (GST) inhibitor 4-Chloro-7-nitro-1,2,3-benzoxadiazole (NBD-Cl) all partially reversed HLJ resistance. Among them, the synergistic effect of PBO and nicosulfuron is the most significant. This is the first report of resistance to nicosulfuron in crabgrass through ALS gene overexpression and possible metabolic resistance.

Network pharmacology, molecular docking-driven, Qbd-Engineered antifungal in-situ gel loaded with voriconazole nanostructured lipid carriers

Fungal infections (FIs) affect majority of the population, but the current treatments face challenges in terms of their effectiveness. This study focused on specific fungal targets, including dihydrofolate reductase (DHFR), acetohydroxy-acid synthase (AHAS), farnesyltransferase and endoglucanase. The docking studies were conducted with the drug voriconazole (VCZ), comparing it with Fluconazole (FCZ) and Amphotericin B (ATB) against 11 protein data bank (PDB) IDs (IDYR, 3NZB, 6DEQ, 1KS5, 7T0C, 1FY4, 5AJH, 7R79, 6TZ6 and 6IDY). Molecular dynamics (MD) analysis, including RMSD, RMSF, PCA and FEL, confirmed the stability of VCZ. The solubility of VCZ was a problem, so nanostructured lipid carriers (NLCs) were developed to improve ocular penetration. VCF5 was the optimized formulation by using 32 full factorial design. VCZF5-NLCs were the best in terms of nanoparticle size (126.6 nm), Zeta potential (33.5 mV), drug content (DC; 97.38 ± 0.210), encapsulation efficiency (EE; 88.01 ± 0.272) and extended drug release. The results of the ex-vivo corneal diffusion study indicate that VCZ-NLC-loaded in-situ gel (VCZ-NLC-IG3) exhibited DC of 88.25% and drug entrapment (DE) of 74.2%. The results of the zone of inhibition indicated that VCZ-NLC-IG3 had superior efficacy compared to ATB. Network pharmacology showed VCZ interacts with the genes which are responsible for fungus ergosterol biosynthesis, including lanosterol 14-alpha demethylase inhibitors (ERG11), ergosterol biosynthesis protein 5 (ERG5), dimethylallyltransferase 2 (DIT2), ketosynthase (KCN), methylsterol monooxygenase (MSMO1), lamin B receptor (LBR), squalene epoxidase (SQLE), 3-hydroxy-3-methylglutaryl-coenzyme A Reductase (MGCR), 3-hydroxy-3-methylglutaryl-coenzyme A Synthase (HMGCS) and 3-keto-steroid reductase (HSD17B7). In conclusion, the optimized VCZ-loaded NLCs present a promising approach to treat ocular FIs. Communicated by Ramaswamy H. Sarma

Target gene mutations endowed cross-resistance to acetolactate synthase-inhibiting herbicides in wild Brassica juncea

Wild Brassica juncea is a troublesome weed that infests wheat fields in China. Two suspected wild B. juncea populations (19–5 and 19–6) resistant to acetolactate synthase (ALS) inhibitors were collected from wheat fields in China. To clarify their resistance profiles and resistance mechanism, the resistance levels of populations 19–5 and 19–6 to ALS-inhibiting herbicides and their underlying target-site resistance mechanism were investigated. The results showed that the 19–5 population exhibited resistance to tribenuron-methyl, pyrithiobac‑sodium and florasulam, while the 19–6 population was resistant to tribenuron-methyl, pyrithiobac‑sodium, imazethapyr and florasulam. Using the homologous cloning method, two ALS genes were identified in wild B. juncea, with one gene (ALS1) encoding 652 amino acids and the other (ALS2) encoding 655 amino acids. Pro-197-Arg mutation on ALS2 and Trp-574-Leu mutation on ALS1, together with the combination of these two mutations in a single plant, were observed in both 19–5 and 19–6 populations. ALS2 enzymes carrying the Pro-197-Arg mutation were cross-resistant to tribenuron-methyl, pyrithiobac‑sodium, imazerthapyr and florasulam, with resistance index (RI) values of 6.23, 32.81, 7.97 and 1162.50, respectively. Similarly, ALS1 enzymes with Trp-574-leu substitutions also displayed high resistance to these four herbicides (RI values ranging from 132.61 to 3375.00). In addition, the combination of Pro-197-Arg (ALS2) and Trp-574-Leu (ALS1) mutations increased the resistance level of the ALS enzyme to ALS inhibitors, with its RI values 3.83–214.19, 6.88–37.34, 1.91–31.82 and 2.03–5.90-fold higher than a single mutation for tribenuron-methyl, pyrithiobac‑sodium, imazerthapyr and florasulam, respectively. Collectively, Pro-197-Arg mutation on ALS2, Trp-574-Leu mutation on ALS1 and the combination of Pro-197-Arg (ALS2) and Trp-574-Leu (ALS1) mutations in wild B. juncea could endow broad-spectrum resistance to ALS inhibitors, which might provide guides for establishing effective strategies to prevent or delay such resistance evolution in this weed.

A high diversity of non-target site resistance mechanisms to acetolactate-synthase (ALS) inhibiting herbicides has evolved within and among field populations of common ragweed (Ambrosia artemisiifolia L.)

BackgroundNon-target site resistance (NTSR) to herbicides is a polygenic trait that threatens the chemical control of agricultural weeds. NTSR involves differential regulation of plant secondary metabolism pathways, but its precise genetic determinisms remain fairly unclear. Full-transcriptome sequencing had previously been implemented to identify NTSR genes. However, this approach had generally been applied to a single weed population, limiting our insight into the diversity of NTSR mechanisms. Here, we sought to explore the diversity of NTSR mechanisms in common ragweed (Ambrosia artemisiifolia L.) by investigating six field populations from different French regions where NTSR to acetolactate-synthase-inhibiting herbicides had evolved.ResultsA de novo transcriptome assembly (51,242 contigs, 80.2% completeness) was generated as a reference to seek genes differentially expressed between sensitive and resistant plants from the six populations. Overall, 4,609 constitutively differentially expressed genes were identified, of which none were common to all populations, and only 197 were shared by several populations. Similarly, population-specific transcriptomic response was observed when investigating early herbicide response. Gene ontology enrichment analysis highlighted the involvement of stress response and regulatory pathways, before and after treatment. The expression of 121 candidate constitutive NTSR genes including CYP71, CYP72, CYP94, oxidoreductase, ABC transporters, gluco and glycosyltransferases was measured in 220 phenotyped plants. Differential expression was validated in at least one ragweed population for 28 candidate genes. We investigated whether expression patterns at some combinations of candidate genes could predict phenotype. Within populations, prediction accuracy decreased when applied to an additional, independent plant sampling. Overall, a wide variety of genes linked to NTSR was identified within and among ragweed populations, of which only a subset was captured in our experiments.ConclusionOur results highlight the complexity and the diversity of NTSR mechanisms that can evolve in a weed species in response to herbicide selective pressure. They strongly point to a non-redundant, population-specific evolution of NTSR to ALS inhibitors in ragweed. It also alerts on the potential of common ragweed for rapid adaptation to drastic environmental or human-driven selective pressures.

Characterization of mesosulfuron-methyl + iodosulfuron-methyl and pyroxsulam–resistant annual bluegrass (Poa annua) in an annual cropping system

AbstractHerbicide-resistant annual bluegrass (Poa annua L.) has become a problem in non-arable land areas. In arable fields, P. annua is frequently of lower priority in weed control program due to the variety of control options available and the relatively modest impact on crop yield compared with other species. In Ireland, postemergence herbicides are not primarily intended for P. annua control, but some herbicides, including the acetolactate synthase (ALS) inhibitor mesosulfuron-methyl + iodosulfuron-methyl, exhibit P. annua activity. In this study, a suspected P. annua population (POAAN-R) that survived mesosulfuron-methyl + iodosulfuron-methyl at 0.75 of the field recommended rate was sampled from a wheat (Triticum aestivum L.) field in County Dublin, Ireland. Single-dose testing confirmed that the suspected POAAN-R had evolved resistance to mesosulfuron-methyl + iodosulfuron-methyl and, additionally, to pyroxsulam (not registered in Ireland for P. annua control), but was sensitive to clethodim, glyphosate, pendimethalin, and flufenacet. Dose–response experiments indicated that POAAN-R was more resistant (GR50 resistance index) to both mesosulfuron-methyl + iodosulfuron-methyl (47.8 times) and pyroxsulam (38.0 times) than sensitive POAAN-S, and this was associated with the mutation at Trp-574 in the ALS protein. Malathion (a cytochrome P450 [P450] inhibitor) pretreatment did not reverse POAAN-R resistance to mesosulfuron-methyl + iodosulfuron-methyl or pyroxsulam at the field rate or above. The natural inherent mutation at Ile-1781 in acetyl-CoA carboxylase protein had no effect on both POAAN-R and POAAN-S sensitivity to clethodim. The glyphosate sensitivity of POAAN-R also corresponded with no known mutation in 5-enolpyruvylshikimate-3-phosphate synthase protein. Based on field histories, poor early-season weed control coupled with intensive use of mesosulfuron-methyl + iodosulfuron-methyl (often at reduced rates) has unintentionally selected for ALS inhibitor–resistant POAAN-R. This is the first report to characterize resistance in P. annua to ALS-inhibiting herbicides mesosulfuron-methyl + iodosulfuron-methyl and pyroxsulam in an arable setting. There is an opportunity to effectively control POAAN-R using herbicides, but this needs a wide-ranging and varied approach, coupled with cultural/nonchemical practices.

Amaranthus palmeri S. Watson: A new threat to agriculture in Europe and the Mediterranean region

AbstractAmaranthus palmeri S. Watson is a dioecious broad‐leaved species characterised by rapid growth, high seed production, efficient resource use and adaptability to diverse environments and cropping systems. A. palmeri populations are spreading beyond their native range in south western United States, into the Mediterranean region and have been reported in several European and Mediterranean countries. First stands were located along roadsides, and later in nearby agricultural fields. Possible routes of introduction include contaminated animal feed and grain shipments for industrial processing. Some European A. palmeri populations appear to be already resistant to 5‐enolpyruvylshikimate‐3‐phosphate synthase and acetolactate synthase inhibitors when imported. Recent studies suggest that this trait may have evolved in the origin country before the introduction into southern Europe and the wider Mediterranean region. Effective, transboundary networks have to be established between farmers and researchers to raise agricultural community awareness and continuously monitor the invasiveness of A. palmeri in Europe. In this review, we first highlight botanical characteristics that allow accurate identification of A. palmeri from closely related Amaranthus species. Then, we summarise recent reports on the occurrence of A. palmeri in Europe and the Mediterranean region. We proceed to assess future invasion and spread across Europe according to climate and agricultural practices. In addition, we review measures to early detect A. palmeri and emphasised on feasible, chemical and non‐chemical weed control methods as components of integrated weed management to mitigate the impact of A. palmeri invasion on European agriculture.

Prevalence, spatial structure and evolution of resistance to acetolactate-synthase (ALS) inhibitors and 2,4-D in the major weed Papaver rhoeas (L.) assessed using a massive, country-wide sampling.

Corn poppy (Papaver rhoeas) is the most damaging broadleaf weed in France. Massively parallel amplicon sequencing was used to investigate the prevalence, mode of evolution and spread of resistance-endowing ALS alleles in 422 populations randomly sampled throughout poppy's range in France. Bioassays were used to detect resistance to the synthetic auxin 2,4-D in 43 of these populations. A total of 21 100 plants were analysed and 24 mutant ALS alleles carrying an amino-acid substitution involved or potentially involved in resistance were identified. The vast majority (97.6%) of the substitutions occurred at codon Pro197, where all six possible single-nucleotide non-synonymous substitutions plus four double-nucleotide substitutions were identified. Changes observed in the enzymatic properties of the mutant ALS isoforms could not explain the differences in prevalence among the corresponding alleles. Sequence read analysis showed that mutant ALS alleles had multiple, independent evolutionary origins, and could have evolved several times independently within an area of a few kilometres. Finally, 2,4-D resistance was associated with mutant ALS alleles in individual plants in one third of the populations assayed. The intricate geographical mosaic of mutant ALS alleles observed is the likely result of the combination of huge population sizes, multiple independent mutation events and human-mediated spread of resistance. Our work highlights the ability of poppy populations and individual plants to accumulate different ALS alleles and as yet unknown mechanisms conferring resistance to synthetic auxins. This does not bode well for the continued use of chemical herbicides to control poppy. © 2023 Society of Chemical Industry.

Structure-activity relationships of bensulfuron methyl and its derivatives as novel agents against drug-resistant Candida auris.

With the emergence of the human pathogen Candida auris as a threat to human health, there is a strong demand to identify effective medicines to prevent the harm caused by such drug-tolerant human fungi. Herein, a series of 33 new derivatives of bensulfuron methyl (BSM) were synthesized and characterized by 1 H NMR, 13 C NMR, and HRMS. Among the target compounds, 8a possessed the best Ki value of 1.015 μM against C. auris acetohydroxyacid synthase (CauAHAS) and an MIC value of 6.25 μM against CBS10913, a clinically isolated strain of C. auris. Taken together the structures of BSM and the synthesized compounds, it was found that methoxy groups at both meta-position of pyrimidine ring are likely to provide desirable antifungal activities. Quantum calculations and molecular dockings were performed to understand the structure-activity relationships. The present study has hence provided some interesting clues for the discovery of novel antibiotics with this distinct mode of action.