Herbicidal activity on acetolactate synthase‐resistant barnyardgrass (Echinochloa crus‐galli) in Arkansas,USA

The intensive use of the acetolactate synthase (ALS)‐inhibiting herbicides, imazethapyr, penoxsulam and bispyribac‐sodium, in imidazolinone‐resistant (Clearfield) rice increases the risk of the evolution of ALS‐resistant barnyardgrass. In 2009, imazethapyr failed to control barnyardgrass that was collected from a field in Arkansas, USA, following the failure of the herbicide in 2008. A greenhouse experiment was conducted to confirm and document the level of resistance of the biotype against three ALS‐inhibiting herbicides that currently are labeled in rice. The level of control of the resistant biotype at the labeled rate of bispyribac‐sodium of 35 g ai ha^?1 was 10%, penoxsulam at 22 g ai ha^?1 was 0% and imazethapyr at 70 g ai ha^?1 was 25%. The level of mortality of the susceptible biotype was 100% with all the herbicides at the labeled rate. The dose needed to kill 50% of the resistant plants was 49 g ha^?1 of bispyribac‐sodium, 254 g ha^?1 of penoxsulam and 170 g ha^?1 of imazethapyr. For the susceptible biotype, bispyribac‐sodium at 6 g ha^?1, penoxsulam at 10 g ha^?1 and imazethapyr at 12 g ha^?1 killed 50% of the treated plants. Based on these findings, it was confirmed that a barnyardgrass population has evolved cross‐resistance to three ALS‐inhibiting herbicides in rice culture in Arkansas. Furthermore, an experiment was conducted to determine if the ALS‐resistant biotype could be controlled using other mechanisms of action. The results indicated that propanil, a photosystem II inhibitor, and quinclorac, a synthetic auxin, failed to control the resistant biotype at the labeled rates, whereas all the other evaluated herbicides provided effective control of both biotypes.     

河南省多花黑麦草对ACCase和ALS抑制剂的抗性及其靶标基因突变分析

为明确河南省部分地区的多花黑麦草Lolium multiflorum种群对乙酰辅酶A羧化酶（acetylCoA carboxylase,ACCase）和乙酰乳酸合成酶（acetolactate synthase,ALS）抑制剂类除草剂的抗性水平和抗性机理,采用整株生物测定法测定采自新乡市和驻马店市的多花黑麦草种群对ACCase抑制剂类除草剂精噁唑禾草灵、炔草酯、唑啉草酯和ALS抑制剂类除草剂甲基二磺隆、氟唑磺隆、啶磺草胺的抗性水平,并对多花黑麦草ACCase和ALS靶标酶编码基因进行克隆及氨基酸序列比对,分析其靶标抗性机理。结果显示,与多花黑麦草敏感种群HNXX01相比,HNZMD04和HNXX05种群对6种除草剂均产生了抗性,HNZMD04种群对精噁唑禾草灵和啶磺草胺的相对抗性倍数分别为44.65和40.31,对炔草酯和氟唑磺隆的相对抗性倍数分别为11.91和11.93;HNXX05种群对精噁唑禾草灵和氟唑磺隆的相对抗性倍数分别为27.70和25.67。HNZMD04和HNXX05抗性种群的ACCase基因均发生了D2078G突变,2个种群的突变率分别为55%和70%;HNZMD04...     

Corn poppy (Papaver rhoeas) cross-resistance to ALS-inhibiting herbicides

BACKGROUND: Papaver rhoeas (L.) has evolved resistance to tribenuron in winter wheat fields in northern Greece owing to multiple Pro₁₉₇ substitutions. Therefore, the cross‐resistance pattern to other sulfonylurea and non‐sulfonylurea ALS‐inhibiting herbicides of the tribenuron resistant (R) and susceptible (S) corn poppy populations was studied by using whole‐plant trials and in vitro ALS catalytic activity assays. RESULTS: The whole‐plant trials revealed that tribenuron R populations were also cross‐resistant to sulfonylureas mesosulfuron + iodosulfuron, chlorsulfuron and triasulfuron. The whole‐plant resistance factors (RFs) calculated for pyrithiobac, imazamox and florasulam ranged from 12.4 to > 88, from 1.5 to 28.3 and from 5.6 to 25.4, respectively, and were lower than the respective tribenuron RF values (137 to > 2400). The ALS activity assay showed higher resistance of the ALS enzyme to sulfonylurea herbicides (tribenuron > chlorsulfuron) and lower resistance to non‐sulfonylurea ALS‐inhibiting herbicides (pyrithiobac > florasulam ≈ imazamox). CONCLUSION: These findings indicate that Pro₁₉₇ substitution by Ala, Ser, Arg or Thr in corn poppy results in a less sensitive ALS enzyme to sulfonylurea herbicides than to other ALS‐inhibiting herbicides. The continued use of sulfonylurea herbicides led to cross‐resistance to all ALS‐inhibiting herbicides, making their use impossible in corn poppy resistance management programmes. Copyright © 2011 Society of Chemical Industry     

Cross‐resistance of Echinochloa species to acetolactate synthase inhibitor herbicides

This study was conducted to evaluate the cross‐resistance of acetolactate synthase (ALS) inhibitors with different chemistries, specifically azimsulfuron (sulfonylurea), penoxsulam (triazolopyrimidine sulfonanilide) and bispyribac‐sodium (pyrimidinyl thio benzoate), in Echinochloa oryzicola and Echinochloa crus‐galli that had been collected in South Korea and to investigate their herbicide resistance mechanism. Both Echinochloa spp. showed cross‐resistance to the ALS inhibitors belonging to the above three different chemistries. In a whole plant assay with herbicides alone, the resistant/susceptible ratios for azimsulfuron, penoxsulam and bispyribac‐sodium were 12.6, 28.1 and 1.9 in E. oryzicola and 21.1, 13.7 and 1.8 in E. crus‐galli, respectively. An in vitro ALS enzyme assay with herbicides showed that the I ₅₀‐values of the resistant accessions were approximately two‐to‐three times higher than the susceptible accessions, with no statistical difference, suggesting that the difference in ALS sensitivity cannot explain ALS inhibitor resistance in Echinochloa spp. for azimsulfuron, penoxsulam and bispyribac‐sodium. A whole plant assay with fenitrothion showed that the GR ₅₀‐values significantly decreased in both the resistant E. oryzicola and E. crus‐galli accessions when azimsulfuron, penoxsulam and bispyribac‐sodium were applied with the P450 inhibitor, while no significant decrease was observed in the susceptible accessions when the P450 inhibitor was used. Thus, these results suggest that ALS inhibitor cross‐resistance for azimsulfuron, penoxsulam and bispyribac‐sodium is related to enhanced herbicide metabolism.     

Amaranthus hybridus populations resistant to triazine and acetolactate synthase-inhibiting herbicides

Amaranthus hybridus L. populations (A, B and C) obtained from escapes in Massac County and Pope County fields in southern Illinois, USA were subjected to greenhouse and laboratory experiments to measure multiple resistance to triazine and acetolactate synthase (ALS)‐inhibiting herbicides and cross‐resistance between sulfonylurea and imidazolinone herbicides. Phytotoxicity responses of the three populations revealed that only population B exhibited multiple resistances to triazine and ALS‐inhibiting herbicides. This population was >167‐, >152‐ and >189‐fold resistant to atrazine, imazamox and thifensulfuron, respectively, at the whole plant level compared with the susceptible population. Population A was only resistant to triazines and population C was only resistant to ALS‐inhibiting herbicides. Results from in vivo ALS enzyme and chlorophyll fluorescence assays confirmed these findings and indicated that an altered site‐of‐action mediated resistance to both triazine and ALS‐inhibiting herbicides. Gene sequencing revealed that a glycine for serine substitution at residue 264 of the D1 protein, and a leucine for tryptophan substitution at residue 574 of ALS were the causes of resistance for the three populations.     

抗精噁唑禾草灵和甲基二磺隆看麦娘ACCase和ALS基因突变

为明确看麦娘Alopecurus aequalis抗性种群YL的靶标抗性机制,采用基因克隆法对看麦娘抗性和敏感种群间乙酰辅酶A羧化酶(ACCase)和乙酰乳酸合成酶(ALS)基因序列进行扩增、克隆和测序,比对二者ACCase和ALS基因序列的差异,探寻其产生抗药性突变的基因位点,同时测定该突变型抗性种群YL对不同ACCase和ALS抑制剂类除草剂的交互抗性。结果显示,与看麦娘敏感种群TL相比,抗性种群YL的ACCase基因CT区域第2 041位氨基酸由异亮氨酸(ATT)突变为天冬酰胺酸(AAT),ALS基因Domain A区域第197位氨基酸由脯氨酸(CCC)突变为精氨酸(CGC)。看麦娘抗性种群YL对ACCase抑制剂炔草酯产生了高水平抗性,抗性倍数为43.96,对高效氟吡甲禾灵和精喹禾灵产生了中等水平抗性,抗性倍数分别为18.33和15.87,对唑啉草酯、烯草酮和烯禾啶较敏感;对ALS抑制剂氟唑磺隆产生了低水平抗性,抗性倍数为8.39,对啶磺草胺和咪唑乙烟酸较敏感。表明ACCase基因第2 041位和ALS基因第197位氨基酸突变是导致看麦娘抗性种群YL对精噁唑禾草灵和甲基二磺隆同时产生抗性的重要原因之一。     

Mutations of the ALS gene endowing resistance to ALS-inhibiting herbicides in Lolium rigidum populations

BACKGROUND: In the important grass weed Lolium rigidum (Gaud.), resistance to ALS‐inhibiting herbicides has evolved widely in Australia. The authors have previously characterised the biochemical basis of ALS herbicide resistance in a number of L. rigidum biotypes and established that resistance can be due to a resistant ALS and/or enhanced herbicide metabolism. The purpose of this study was to identify specific resistance‐endowing ALS gene mutation(s) in four resistant populations and to develop PCR‐based molecular markers. RESULTS: Six resistance‐conferring ALS mutations were identified: Pro‐197‐Ala, Pro‐197‐Arg, Pro‐197‐Gln, Pro‐197‐Leu, Pro‐197‐Ser and Trp‐574‐Leu. All six mutations were found in one population (WLR1). Each Pro‐197 mutation conferred resistance to the sulfonylurea (SU) herbicide sulfometuron, whereas the Trp‐574‐Leu mutation conferred resistance to both sulfometuron and the imidazolinone (IMS) herbicide imazapyr. A derived cleaved amplified polymorphic sequences (dCAPS) marker was developed for detecting resistance mutations at Pro‐197. Furthermore, cleaved amplified polymorphic sequences (CAPS) markers were developed for detecting each of the six mutant resistant alleles. Using these markers, the authors revealed diverse ALS‐resistant alleles and genotypes in these populations and related them directly to phenotypic resistance to ALS‐inhibiting herbicides. CONCLUSION: This study established the existence of a diversity of ALS gene mutations endowing resistance in L. rigidum populations: 1–6 different mutations were found within single populations. At field herbicide rates, resistance profiles were determined more by the specific mutation than by whether plants were homo‐ or heterozygous for the mutation. Copyright © 2008 Society of Chemical Industry     

Isolation and expression of genes for acetolactate synthase and acetyl-CoA carboxylase in Echinochloa phyllopogon, a polyploid weed species

BACKGROUND: Target‐site resistance is the major cause of herbicide resistance to acetolactate synthase (ALS)‐ and acetyl‐CoA carboxylase (ACCase)‐inhibiting herbicides in arable weeds, whereas non‐target‐site resistance is rarely reported. In the Echinochloa phyllopogon biotypes resistant to these herbicides, target‐site resistance has not been reported, and non‐target‐site resistance is assumed to be the basis for resistance. To explore why target‐site resistance had not occurred, the target‐site genes for these herbicides were isolated from E. phyllopogon, and their expression levels in a resistant biotype were determined. RESULTS: Two complete ALS genes and the carboxyltransferase domain of four ACCase genes were isolated. The expression levels of ALS and ACCase genes were higher in organs containing metabolically active meristems, except for ACC4, which was not expressed in any organ. The differential expression among examined organs was more prominent for ALS2 and ACC2 and less evident for ALS1, ACC1 and ACC3. CONCLUSION: E. phyllopogon has multiple copies of the ALS and ACCase genes, and different expression patterns were observed among the copies. The existence of three active ACCase genes and the difference in their relative expression levels could influence the occurrence of target‐site resistance to ACCase inhibitors in E. phyllopogon. Copyright © 2012 Society of Chemical Industry     

A novel amino acid substitution Ala-122-Tyr in ALS confers high-level and broad resistance across ALS-inhibiting herbicides

BACKGROUND: Wild radish, a problem weed worldwide, is a severe dicotyledonous weed in crops. In Australia, sustained reliance on ALS‐inhibiting herbicides to control this species has led to the evolution of many resistant populations endowed by any of several ALS mutations. The molecular basis of ALS‐inhibiting herbicide resistance in a novel resistant population was studied. RESULTS: ALS gene sequencing revealed a previously unreported substitution of Tyr for Ala at amino acid position 122 in resistant individuals of a wild radish population (WARR30). A purified subpopulation individually homozygous for the Ala‐122‐Tyr mutation was generated and characterised in terms of its response to the different chemical classes of ALS‐inhibiting herbicides. Whole‐plant dose‐response studies showed that the purified subpopulation was highly resistant to chlorsulfuron, metosulam and imazamox, with LD₅₀ or GR₅₀ R/S ratio of > 1024, > 512 and > 137 respectively. The resistance to imazypyr was found to be relatively moderate (but still substantial), with LD₅₀ and GR₅₀ R/S ratios of > 16 and > 7.8 respectively. In vitro ALS activity assays showed that Ala‐122‐Tyr ALS was highly resistant to all tested ALS‐inhibiting herbicides. CONCLUSION: The molecular basis of ALS‐inhibiting herbicide resistance in wild radish population WARR30 was identified to be due to an Ala‐122‐Tyr mutation in the ALS gene. This is the first report of an amino acid substitution at Ala‐122 in the plant ALS that confers high‐level and broad‐spectrum resistance to ALS‐inhibiting herbicides, a remarkable contrast to the known mutation Ala‐122‐Thr endowing resistance to imidazolinone herbicide. Copyright © 2012 Society of Chemical Industry     

Response of an acetolactate synthase (ALS)-resistant biotype of Amaranthus rudis to selected ALS-inhibiting and alternative herbicides

An Amaranthus rudis Saner (common water-hemp) biotype from a field treated for two consecutive years with a mixture of chlorimuron and metribuzin was tested in greenhouse and laboratory studies to assess resistance and cross-resistance to four acetolactale synthase (ALS)-inhibiting herbicides. The biotype demonstrated >1920-fold resistance at the whole plant level, and >850-fold resistance at the ALS enzyme to chlorimuron, compared with a susceptible biotype. This chlorimuron-resistant biotype was also cross-resistant to primisuifu-ron, haiosulfuron and imazethapyr. In greenhouse studies, atrazine alone or in combination with ALS-inhibiting herbicides provided excellent control of the resistant biotype of A. rudis . Combinations of dicamba and ALS-inhibitors also provided adequate control. Additionally, premixtures of flumetsulam and metolachlor and of dicamba and atrazine furnished excellent control of this chlorimuron-resistant A. rudis biotype.     

荠菜对乙酰乳酸合成酶抑制剂类除草剂的抗性水平及其分子机制

为明确荠菜种群对苯磺隆的抗性水平及其靶标抗性产生的分子机制,采用整株水平测定法测定了荠菜对苯磺隆及其他5种乙酰乳酸合成酶（ALS）抑制剂类除草剂的抗性水平,同时扩增和比对了荠菜抗性和敏感种群之间ALS基因的差异。结果显示:与敏感种群15-ZMD-1相比,抗性种群15-ZMD-5对苯磺隆产生了高水平抗性,抗性倍数为219.6;15-ZMD-5种群不同单株中共存在3种突变方式,分别为ALS基因197位点脯氨酸（CCT）突变为亮氨酸（CTT）、574位点色氨酸（TGG）突变为亮氨酸（TTG）以及单株同时发生上述197和574位点的氨基酸突变。15-ZMD-5抗苯磺隆种群对嘧草硫醚、啶磺草胺和氟唑磺隆均产生了高水平的交互抗性,抗性倍数分别为41.2、79.3和87.8;对双氟磺草胺和咪唑乙烟酸产生了低水平的交互抗性,抗性倍数分别为8.5和5.6。分析表明,荠菜抗性种群ALS基因发生的氨基酸突变可能是导致其对ALS抑制剂类除草剂产生抗性的重要原因之一。     

Amaranthus palmeri resistance and differential tolerance of Amaranthus palmeri and Amaranthus hybridus to ALS-inhibitor herbicides

Suspected imazaquin-resistant accessions of Amaranthus palmeri were studied to determine the magnitude of resistance and cross-resistance to acetolactate synthase (ALS)-inhibiting herbicides and compare differential tolerance of A palmeri and Amaranthus hybridus to ALS inhibitors. Five of seven A palmeri accessions were resistant to imazaquin. The most imazaquin-resistant accession, accession 7, also showed 74, 39 and 117 times higher resistance than the susceptible biotype to chlorimuron, diclosulam and pyrithiobac, respectively. Resistance to imazaquin and cross-resistance to other ALS inhibitors in A palmeri was due to a less-sensitive ALS enzyme. A palmeri was 70 times more tolerant to imazaquin than A hybridus. A palmeri was also seven times more tolerant to pyrithiobac than A hybridus. Differences in ALS enzyme sensitivity could not fully account for the high tolerance of A palmeri to imazaquin compared to A hybridus. Both species were equally affected by chlorimuron and diclosulam.     

Resistance levels of sulfonylurea‐resistant Schoenoplectus juncoides (Roxb.) Palla with various Pro197 mutations in acetolactate synthase to imazosulfuron,bensulfuron‐methyl,metsulfuron‐methyl and imazaquin‐ammonium

An investigation, using herbicidal pot tests in a greenhouse condition, was conducted to determine the whole‐plant dose–response relationships to several acetolactate synthase (ALS)‐inhibiting herbicides of sulfonylurea (SU)‐resistant Schoenoplectus juncoides with various Pro₁₉₇ mutations in ALS that was collected from Japanese rice paddy fields. All the tested SU‐resistant accessions with a Pro₁₉₇ mutation were highly resistant to two commonly used SU herbicides (imazosulfuron and bensulfuron‐methyl), but were much less resistant to another SU herbicide, metsulfuron‐methyl, and were substantially not resistant to imazaquin‐ammonium. These cross‐resistance patterns have been known previously in fragments of S. juncoides and other weed species and were comprehensively confirmed in this study with a whole set of Pro₁₉₇ mutations. The analyses of resistance levels, based on ED₉₀ values, newly showed that different accessions with a common amino acid substitution in ALS1 showed similar responses to these herbicides (confirmed with four amino acid substitutions), that the rankings of resistance levels that were conferred by various Pro₁₉₇ mutations in ALS1 differed among the SU herbicides and that the resistance levels of the ALS2‐mutated accessions were higher than, lower than or similar to those of the corresponding ALS1‐mutated accessions, depending on the compared pair, but the deviation patterns were generally similar among the SU herbicides in each compared pair. The final finding might suggest that the abundance of ALS2 is not as stable as that of ALS1. In addition, as a result of these new findings, together with expected further research, a suggested possibility is that substituting amino acids at Pro₁₉₇ generally could be estimated by plotting each accession's ED₉₀ values of imazosulfuron and bensulfuron‐methyl in a two‐dimensional graph.     

Biology and mechanisms of sulfonylurea resistance in Schoenoplectiella juncoides,a noxious sedge in the rice paddy fields of Japan

Schoenoplectiella juncoides is a noxious sedge weed in rice paddy fields that has evolved resistance to sulfonylurea (SU) herbicides. The molecular basis of resistance is amino acid substitutions at Pro₁₉₇, Trp₅₇₄ or Asp₃₇₆ in the acetolactate synthase (ALS) enzyme, which is the target of SUs. Schoenoplectiella juncoides has two ALS genes and resistant plants have point mutations that cause amino acid substitutions in either encoded protein. Single‐nucleotide substitutions at the codon for Pro₁₉₇ in the ALS genes can cause six types of amino acid substitutions and all of these substitutions have been found in both ALS genes among Japanese SU‐resistant biotypes. Whole‐plant herbicide responses differ among the amino acid substitution types. Furthermore, analyses of ALS activity in plant extracts show that the extracts’ responses to herbicides differ, depending on which ALS gene is mutated. The activity responses of the ALS extracts to the SU, imazosulfuron, showed double‐sigmoid curves with plateaus of ~30% inhibition for Pro₁₉₇ substitutions in ALS1 and ~70% for Pro₁₉₇ substitutions in ALS2. This indicates that ALS1 and ALS2 contribute to the responses with a proportion of 7:3. The double‐sigmoid curves can be reconstructed to show the responses of the resistant and susceptible enzymes separately by regression analysis. The resistance levels of the separate ALS1 or ALS2 mutated enzyme are highly correlated with the whole‐plant responses, with a relationship that the former is the square of the latter. This could provide a quantitative insight into the physiological basis of resistance.     

Mechanisms of resistance to acetyl‐coenzyme A carboxylase‐inhibiting herbicides in a Hordeum leporinum population

A failure of acetyl‐coenzyme A carboxylase (ACCase)‐inhibiting herbicides to control a population of Hordeum leporinum Link (barleygrass) occurred following eight applications of these herbicides in both crops and pastures. This population was 7.6‐fold resistant to fluazifop‐P‐butyl compared with standard susceptible populations. The population was between 3.6‐ and 3.8‐fold resistant to other ACCase‐inhibiting herbicides, except butroxydim to which it was susceptible. ACCase extracted from resistant plants and assayed in the presence of herbicides in vitro was susceptible to fluazifop acid and other aryloxyphenoxypropanoate herbicides, but was 4‐fold less sensitive to sethoxydim compared with ACCase from susceptible plants. Resistant plants metabolised fluazifop acid about 1.3‐fold more rapidly compared with susceptible plants; however, sethoxydim was metabolised equally in both populations. Resistance to fluazifop‐P‐butyl and other aryloxyphenoxypropanoate herbicides may be the result of increased herbicide detoxification, whereas resistance to sethoxydim appears to be due to a modified target enzyme. Herbicide resistance in this population is unusual in that different mechanisms appear to confer resistance to the aryloxyphenoxypropanoate and cyclohexanedione herbicides. © 2000 Society of Chemical Industry     

Varied occurrence of diverse sulfonylurea‐resistant biotypes of Schoenoplectus juncoides [Roxb.] Palla in Japan,as classified by an acetolactate synthase gene mutation

Suspected sulfonylurea (SU)‐resistant Schoenoplectus juncoides plants were collected from rice paddy fields at 24 sites in Japan in order to discover the occurrence pattern of target‐site substitutions on a nationwide scale and at a local field scale. A genetic analysis of the two acetolactate synthase (ALS) genes, ALS1 and ALS2, of the collected plants confirmed that a single‐nucleotide mutation at the Pro₁₉₇, Asp₃₇₆ or Trp₅₇₄ site of either ALS1 or ALS2 existed in each suspected SU‐resistant plant. On a nationwide scale, it was shown that the ALS1 mutations and the ALS2 mutations occurred at a similar frequency, that the P197S and the P197L substitutions were found most frequently among all the substitutions, and that the W574L substitutions (known as global resistance to any ALS‐inhibiting herbicide) were found at a relatively low frequency but in a geographically wide range. In the local field‐scale survey, which was conducted at two sites in Hyogo Prefecture, it was shown that the substitutions were less diverse, compared to on a nationwide scale, probably because the investigation involved a limited number of local fields, and that several substitutions and a susceptible biotype were found in single fields suggesting that a number of collections is required in order to understand the local SU‐resistant status of S. juncoides. In addition, this study reported new findings, that of the P197R, P197T and D376E substitutions in S. juncoides. This set of diverse substitutions in a weed species can be used for further research purposes.     

Non-target-site-based resistance to ALS-inhibiting herbicides in six Bromus rigidus populations from Western Australian cropping fields

BACKGROUND: Bromus rigidus is a common weed species that has increased in cropping fields owing to limited control options. During a random field survey in Western Australia, six B. rigidus populations that had survived in‐crop weed control programmes were collected. The study aimed to determine the resistance profile of these six populations. RESULTS: Based on dose–response studies, all six B. rigidus populations had a low‐level resistance to sulfosulfuron and sulfometuron (both sulfonylurea herbicides) while remaining susceptible to herbicides with other modes of action. ALS in vitro activity assays revealed no differences in enzyme sensitivity between susceptible and resistant populations, while the use of malathion (a cytochrome P450 inhibitor) in combination with sulfosulfuron caused the resistant populations to behave like the susceptible population. CONCLUSION: This study established that these six B. rigidus populations have a low‐level resistance to the ALS‐inhibiting sulfonylurea herbicides, but are able to be controlled by other herbicide modes of action. The low‐level, malathion‐reversible resistance, together with a sensitive ALS, strongly suggest that a non‐target‐site enhanced metabolism is the mechanism of resistance. Copyright © 2012 Society of Chemical Industry     

除草剂靶酶-AHAS酶及基因突变体与除草剂设计(Ⅱ).AHAS及W464突变酶与除草活性分子的相互作用

乙酰羟基酸合成酶(AHAS)是磺酰脲类、咪唑啉酮类、三唑嘧啶磺酰胺类及水杨酸类除草剂的作用靶标,大田使用中杂草对这几类除草剂产生抗性的主要因素是AHAS酶的突变。利用大肠杆菌AHAS Ⅱ中464位的色氨酸突变体(W464A、W464F、W464L、W464Y),研究了野生型和突变酶对商品化除草剂(氯嘧磺隆、氯磺隆、咪唑乙烟酸、咪唑喹啉酸)以及烷硫基磺酰脲的敏感性。野生型E. coli AHAS Ⅱ对这些化合物的抑制作用较为敏感,而突变酶对其呈现出不同程度的抗性,使商品化除草剂的抑制常数增加了10~1.0×104倍不等,烷硫基磺酰脲的抑制常数增加幅度较小。烷硫基磺酰脲 1a 对W464L突变酶的高抑制活性,暗示着发展针对靶酶抗性的除草剂的可能性。     

A European biotype of Amaranthus retroflexus cross-resistant to ALS inhibitors and response to alternative herbicides

An acetolactate synthase (ALS)‐resistant Amaranthus retroflexus biotype was collected in a soyabean crop after repeated exposure to imazethapyr and thifensulfuron‐methyl in north‐eastern Italy. Studies were conducted to characterise the resistance status and determine alternative post‐emergence herbicides for controlling this biotype. Whole‐plant bioassay revealed that the GR₅₀ values were 1898‐ and 293‐fold higher than those observed for the biotype susceptible to imazethapyr and imazamox respectively. The biotype also displayed high cross‐resistance to sulfonylureas. Molecular analysis demonstrated that a single nucleotide substitution had occurred in domain B (TGG to TTG at position 574), conferring a change from the amino acid tryptophan to leucine in the resistant biotype. However, herbicides with other modes of action (PSII, 4‐HPPD and PPO inhibitors) provided excellent control. The GR₅₀ ratios for metribuzin, terbuthylazine and mesotrione were close to 1 and treatments with fomesafen gave 100% control of both susceptible and resistant biotypes at the recommended field dose. This study documents the first case of an imidazolinone and ALS‐resistant biotype in European crops and identifies the post‐emergence herbicide options available for managing this troublesome weed in soyabean crops. Alternative management strategies are also discussed.