Mechanism of resistance to fenoxaprop in Japanese foxtail (Alopecurus japonicus) from China

Japanese foxtail is one of the most common and troublesome weeds infesting cereal and oilseed rape fields in China. Repeated use during the last three decades of the ACCase-inhibiting herbicide fenoxaprop-P-ethyl to control this weed has resulted in the occurrence of resistance. Dose–response tests established that a population (AHFD-1) from eastern China had evolved high-level resistance to fenoxaprop-P-ethyl. Based on the resistance index, this resistant population of A. japonicus is 60.31-fold resistant to fenoxaprop-P-ethyl. Subsequently, only a tryptophan to cysteine substitution was identified to confer resistance to fenoxaprop-P-ethyl in this resistant population. ACCase activity tests further confirmed this substitution was linked to resistance. This is the first report of the occurrence of Trp-2027-Cys substitution of ACCase in A. japonicus. From whole-plant pot dose–response tests, we confirmed that this population conferred resistance to other APP herbicides, including clodinafop-propargyl, fluazifop-P-butyl, quizalofop-P-ethyl, haloxyfop-R-methyl, cyhalofop-butyl, metamifop, DEN herbicide pinoxaden, but not to CHD herbicides clethodim, sethoxydim. There was also no resistance observed to ALS-inhibiting herbicides sulfosulfuron, mesosulfuron-methyl, flucarbazone-sodium, pyroxsulam, Triazine herbicide prometryne and glyphosate. However, this resistant population was likely to confer slightly (or no) resistant to Urea herbicides chlortoluron and isoproturon.     

Allelic variation of the ACCase gene and response to ACCase-inhibiting herbicides in pinoxaden-resistant Lolium spp

BACKGROUND: The repeated use of acetyl‐coenzyme A carboxylase (ACCase) inhibiting herbicides to control grass weeds has selected for resistance in Lolium spp. populations in Italy. The efficacy of pinoxaden, a recently marketed phenylpyrazoline herbicide, is of concern where resistance to ACCase inhibitors has already been ascertained. ACCase mutations associated with pinoxaden resistance were investigated, and the cross‐resistance pattern to clodinafop, haloxyfop, sethoxydim, clethodim and pinoxaden was established on homo/heterozygous plants for four mutant ACCase alleles. RESULTS: Seven different mutant ACCase alleles (1781‐Leu, 1999‐Leu, 2041‐Asn, 2041‐Val, 2078‐Gly, 2088‐Arg and 2096‐Ala) and 13 combinations with two types of mutation were detected in the pinoxaden‐resistant plants. The 1781‐Leu allele appears to confer a dominant resistance to pinoxaden, clodinafop, haloxyfop, sethoxydim and clethodim at 60 g AI ha^?1. The 2041‐Asn and 2041‐Val alleles are associated with dominant or partially dominant resistance to FOPs, no substantial resistance to DIMs and a moderate resistance to pinoxaden. The 2088‐Arg allele endows a partially dominant resistance to clodinafop, sethoxydim and most likely to pinoxaden. In addition, non‐target‐site resistance mechanisms seem to be involved in pinoxaden resistance. CONCLUSION: Almost all the ACCase mutations selected in the field by other ACCase inhibitors are likely to confer resistance to pinoxaden. Although pinoxaden is sometimes able to control FOP‐resistant populations, it should not be considered as a sustainable ACCase resistance management tool. The presence of non‐ACCase‐based resistance mechanisms that could confer resistance to herbicides with different modes of action further complicates the resistance management strategies. Copyright © 2011 Society of Chemical Industry     

Cross-resistance patterns to ACCase-inhibiting herbicides conferred by mutant ACCase isoforms in Alopecurus myosuroides Huds. (black-grass), re-examined at the recommended herbicide field rate

BACKGROUND: Target‐site‐based resistance to acetyl‐CoA carboxylase (ACCase) inhibitors in Alopecurus myosuroides Huds. is essentially due to five substitutions (Isoleucine‐1781‐Leucine, Tryptophan‐2027‐Cysteine, Isoleucine‐2041‐Asparagine, Aspartate‐2078‐Glycine, Glycine‐2096‐Alanine). Recent studies suggested that cross‐resistance patterns associated with each mutation using a seed‐based bioassay may not accurately reflect field resistance. The authors aimed to connect the presence of mutant ACCase isoform(s) in A. myosuroides with resistance to five ACCase inhibitors (fenoxaprop, clodinafop, haloxyfop, cycloxydim, clethodim) sprayed at the recommended field rate. RESULTS: Results from spraying experiments and from seed‐based bioassays were consistent for all mutant isoforms except the most widespread, Leucine‐1781. In spraying experiments, Leucine‐1781 ACCase conferred resistance to clodinafop and haloxyfop. Some plants containing Leucine‐1781 or Alanine‐2096 ACCase, but not all, were also resistant to clethodim. CONCLUSION: Leucine‐1781, Cysteine‐2027, Asparagine‐2041 and Alanine‐2096 ACCases confer resistance to fenoxaprop, clodinafop and haloxyfop at field rates. Leucine‐1781 ACCase also confers resistance to cycloxydim at field rate. Glycine‐2078 ACCase confers resistance to all five herbicides at field rates. Only Glycine‐2078 ACCase confers clethodim resistance under optimal application conditions. It may be that Leucine‐1781 and Alanine‐2096 ACCases may also confer resistance to clethodim in the field if the conditions are not optimal for herbicide efficacy, or at reduced clethodim field rates. Copyright © 2008 Society of Chemical Industry     

直播稻田牛筋草对乙酰辅酶A羧化酶类除草剂抗性水平及其分子机制

为明确直播稻田牛筋草对乙酰辅酶A羧化酶 (ACCase) 类除草剂的抗药性水平及其抗性产生的分子机制，采用整株生物测定法测定了牛筋草对6种ACCase类除草剂的抗性水平，并分别对抗性种群和敏感种群的ACCase基因部分片段进行了扩增和测序。结果表明:疑似抗性种群SJ-1对唑酰草胺、氰氟草酯、精唑禾草灵、高效氟吡甲禾灵和烯禾啶产生了高水平抗性，其抗性倍数分别为56.6、62.5、128、52.0和16.3；对烯草酮产生了低水平抗性，相对抗性倍数为4.86。将抗性种群和敏感种群的ACCase基因片段序列进行比对分析发现，SJ-1种群ACCase基因2078位氨基酸由天冬氨酸 (GAT) 突变为甘氨酸 (GGT)，该位点氨基酸突变可能是其对ACCase类除草剂产生抗药性的主要原因之一。     

Cross‐resistance patterns to acetyl coenzyme A carboxylase (ACCase) inhibitors associated with different ACCase mutations in Beckmannia syzigachne

Beckmannia syzigachne (American sloughgrass) is a competitive grass weed found in China. Fenoxaprop‐P‐ethyl is widely used for control of this species in China. Resistance to fenoxaprop‐P‐ethyl in B. syzigachne has been reported to be conferred by an isoleucine(Ile)‐1781‐leucine(Leu) substitution in the gene encoding the herbicide target, acetyl‐CoA carboxylase (ACCase). In this study, three mutations were detected by derived cleaved amplified polymorphic sequence (dCAPS) method in fenoxaprop‐P‐ethyl‐resistant B. syzigachne populations: Ile‐1781‐Leu in population JCWL‐R, Ile‐2041‐Asn in JCJT‐R and Gly‐2096‐Ala in JYJD‐R. The data indicated they were genetically homogeneous (homozygous mutant) at the ACCase locus. The use of cytochrome P450 inhibitors was shown to slightly reduce the GR₅₀ value of fenoxaprop‐P‐ethyl‐resistant populations, from which we inferred a combination of target‐site resistance (TSR) and non‐target‐site resistance (NTSR) was involved in fenoxaprop‐P‐ethyl‐resistance. We characterised the cross‐resistance patterns to ACCase inhibitors in B. syzigachne. The plants in the JCWL‐R population were highly resistant to all tested APPs (aryloxyphen‐oxypropionates), sethoxydim and pinoxaden, and moderately resistant to clethodim. The plants in the JCJT‐R population were highly resistant to fluazifop‐P‐butyl, clodinafop‐propargyl, cyhalofop‐butyl, metamifop and pinoxaden; moderately resistant to haloxyfop‐R‐methyl, quizalofop‐P‐ethyl and sethoxydim; and sensitive to clethodim. The plants in the JYJD‐R population were highly resistant to clodinafop‐propargyl, metamifop and pinoxaden; moderately resistant to haloxyfop‐R‐methyl, cyhalofop‐butyl, quizalofop‐P‐ethyl, fluazifop‐P‐butyl and sethoxydim; and sensitive to clethodim. If resistance to ACCase inhibitors is present in B. syzigachne populations in the field, then our results indicate that clethodim should be used. While we demonstrated the cross‐resistance patterns of TSR resulting from three mutations in B. syzigachne, we also demonstrated that NTSR plays a role in resistance, which will complicate weed management.     

耿氏硬草对乙酰辅酶A羧化酶类除草剂抗性水平及分子机制初探

为明确耿氏硬草Pseudosclerochloa kengiana(Ohwi)Tzvel潜在抗性种群对不同乙酰辅酶A羧化酶(ACCase)类除草剂的抗性水平及其靶标抗性的分子机制,采用剂量-反应曲线法测定了耿氏硬草对精鰁唑禾草灵、炔草酯、烯禾啶、烯草酮和唑啉草酯5种ACCase类除草剂的抗性水平,扩增并比对了耿氏硬草抗性和敏感种群间ACCase基因的差异。结果显示:与敏感种群SD-6相比,耿氏硬草种群SD-32对精鰁唑禾草灵、炔草酯、烯禾啶、烯草酮和唑啉草酯产生了不同水平的抗性,抗性倍数分别为16.5、7.5、15.0、4.4和5.7;SD-32种群ACCase基因CT区域的2078位氨基酸基因由GAT突变为GGT,导致天冬氨酸(Asp)被甘氨酸(Gly)取代。分析表明,ACCase基因2078位氨基酸的突变可能是导致耿氏硬草对ACCase类除草剂产生抗性的重要原因之一。     

Altered acetyl-coenzyme A carboxylase confers resistance to clethodim, fluazifop and sethoxydim in Setaria faberi and Digitaria sanguinalis

Summary Populations of Setaria faberi and Digitaria sanguinalis cross-resistant to sethoxydim and fluazifop-P-butyl were identified in a vegetable cropping system in Wisconsin, USA, in 1991 and 1992 respectively. Experiments were conducted with partially purified acetyl-CoA carboxylase (ACCase) to determine whether resistance to sethoxydim and other ACCase inhibitors in S. faberi and D. sanguinalis resulted from altered enzyme activity. Based on I₅₀ values (the herbicide dose that inhibited ACCase activity by 50% compared with untreated ACCase), ACCase of the resistant accession of S. faberi was 4.8-, 10.6- and 319-fold resistant to clethodim, fluazifop-P acid and sethoxydim, respectively, compared with that of the susceptible accession. Similarly, ACCase of the resistant accession of D. sanguinalis was 5.8-, 10.3- and 66-fold resistant to clethodim, fluazifop-P acid and sethoxydim respectively. These results indicated that resistance to ACCase inhibitors in these accessions of S. faberi and D. sanguinalis resulted from an altered ACCase enzyme that confers a very high level of resistance to sethoxydim.     

广东省稻菜轮作区中牛筋草对十种除草剂的抗性水平及靶基因序列分析

为明确广东省稻菜轮作区中牛筋草对10种常用除草剂的抗性水平及抗性分子机制,采用整株生物测定法测定广东省稻菜轮作区内8个牛筋草种群P1～P8对草甘膦、草铵膦和乙酰辅酶A羧化酶(acetyl-CoA carboxylase,ACCase)抑制剂类等10种除草剂的抗性水平,并进一步分析P1和P8种群相关靶标酶基因5-烯醇丙酮酰莽草酸-3-磷酸合酶(5-enolpyruvyl-shikimate-3-phosphate synthase,EPSPS)、谷氨酰胺合成酶(glutamine synthetase,GS)和ACCase的部分功能区序列特征。结果显示,牛筋草P1～P8种群对草甘膦抗性指数为敏感种群的5.9倍～17.7倍,其中P8种群对草甘膦的抗性水平最高;8个种群对草铵膦也产生了不同程度的抗性,抗性指数为敏感种群的2.3倍～14.2倍,其中P1种群抗性最高。牛筋草P1和P8种群均对ACCase抑制剂类除草剂精喹禾灵、氰氟草酯和噁唑酰草胺产生了交互抗性;P1种群ACCase基因在第2 041位氨基酸处发生突变,该突变在牛筋草种群中首次发现;而P8种群ACCase基因则在第2 027位氨基...     

抗精噁唑禾草灵的日本看麦娘ACCase基因突变

为明确日本看麦娘抗性种群对精噁唑禾草灵的抗性水平及抗性产生的分子机制,采用整株水平测定法测定了日本看麦娘对精噁唑禾草灵的抗性水平,扩增和比对了日本看麦娘抗性和敏感种群间乙酰辅酶A羧化酶(acetyl-Co A carboxylase,ACCase)基因的差异。结果显示,与敏感种群AH-7相比,抗性种群AH-25对精噁唑禾草灵的抗性倍数为33.82;AH-25种群ACCase基因CT区域2 078位氨基酸发生了突变,由天冬氨酸GAT突变为甘氨酸GGT;AH-25种群对炔草酯、烯草酮和烯禾啶产生了高水平的抗性,抗性倍数分别为35.66、38.64和29.14,对高效氟吡甲禾灵产生了低水平的抗性,抗性倍数为3.04,对精喹禾灵和唑啉草酯较敏感。表明ACCase基因2 078位氨基酸的突变可能是导致精噁唑禾草灵产生高水平抗性的重要原因。     

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     

4种除草剂与助剂安融乐协同防除紫花苜蓿田马唐的研究

为探究高效氟吡甲禾灵?精吡氟禾草灵?烯草酮?精喹禾灵对紫花苜蓿田老根马唐的防除效果及助剂安融乐对上述4种药剂的增效作用, 在室内测定马唐幼苗和成株对4种除草剂及添加助剂的敏感性, 并在田间验证上述4种除草剂与助剂混用对老根马唐的防效?室内结果表明, 成株马唐对高效氟吡甲禾灵?精吡氟禾草灵?烯草酮?精喹禾灵的耐药性增加明显, 其中精喹禾灵的耐药性增加最大; 高效氟吡甲禾灵?精吡氟禾草灵?烯草酮?精喹禾灵与安融乐混配后, 成株马唐的鲜重GR50(有效成分)较单用上述除草剂分别减少16.6%?31.0%?31.5%?6.2%, 其中烯草酮与安融乐混配增效作用最为明显?田间结果表明, 安融乐的加入能提高上述4种除草剂对老根马唐的防效, 其中烯草酮和安融乐混配对老根马唐的增效最优, 株防效和鲜重防效较单用分别增加10.39和8.58百分点, 并显著提高紫花苜蓿产量?     

Studies on the effect of different cyclohexane-1,3-diones on de-novo fatty acid biosynthesis in poaceae

Cyclohexane-1,3-diones such as the herbicides cycloxydim, sethoxydim, alloxydim and clethodim are known to be specific inhibitors of the plastid-located acetyl-CoA carboxylase (ACCase) in Poaceae, a key enzyme of de-novo fatty acid biosynthesis in higher plants. Using several new cyclohexane-1,3-dione derivatives and known herbicides, the relationships between chemical structure and enzyme inhibition have been studied. The basic cyclohexane-1,3-dione structure was modified at three different positions. These compounds were tested for inhibition of the de novo fatty-acid biosynthesis in test systems of etioplasts isolated from Avena sativa L. and Hordeum vulgare L. seedlings and also for inhibition of the isolated barley ACCase. The I₅₀ values of these cyclohexane-1,3-diones were determined. The influence of the modification of alkyl chains (length and type of substituent) on the degree of ACCase-inhibition is discussed. Several new compounds were found that were about two orders more active than the known herbicides cycloxydim or sethoxydim in the etioplast and ACCase test systems but not necessarily on the level of whole plants.     

除草剂对蚕豆、油菜田小籽虉草的防除效果和安全性评价

小籽虉草Phalaris minor是近年来危害云南中西部地区冬季作物田的恶性杂草。云南冬季作物以阔叶作物为主, 有必要开展在阔叶作物田小籽虉草的化学防除效果研究。本研究在室内和田间测试了108 g/L高效氟吡甲禾灵EC、150 g/L精吡氟禾草灵EC、10%精喹禾灵EC、69 g/L精噁唑禾草灵EW、240 g/L烯草酮EC、12.5%烯禾啶EC、5%唑啉草酯EC等7种阔叶作物田茎叶处理剂对小籽虉草的防效以及对油菜和蚕豆的安全性。室内研究结果表明, 药后30 d, 5%唑啉草酯EC、12.5%烯禾啶EC、10%精喹禾灵EC在推荐剂量作用下对小籽虉草的鲜重防效超过90%, 杂草植株表现明显枯死症状, 并且安全性指数均大于2; 240 g/L烯草酮EC的株防效较差, 鲜重防效也低于50%; 其他药剂的鲜重防效为80%左右。田间药效试验表明, 5%唑啉草酯EC防效最优, 接近95%, 108 g/L高效氟吡甲禾灵EC和12.5%烯禾啶EC等防效能达到90%, 其他药剂的防效也超过80%, 所有药剂对油菜和蚕豆均安全。以上结果表明, 当前大多数阔叶作物田禾本科杂草除草剂对小籽虉草高效, 对作物安全, 控制小籽虉草的备选除草剂品种较多。     

上海市水稻田千金子对3种乙酰辅酶A羧化酶抑制剂的抗性现状及酶突变机制

为明确上海市水稻田千金子对乙酰辅酶A羧化酶 (ACCase) 抑制剂类除草剂的抗性发生情况及可能存在的抗性机制,在上海市千金子发生严重地区的水稻田共采集51个种群,采用单剂量抗性甄别法测定了不同千金子种群对3种ACCase抑制剂类除草剂的抗性水平,扩增和比对了靶标酶ACCase基因部分片段的差异。结果显示:在氰氟草酯105 g/hm2有效成分剂量选择压下,8个千金子种群标记为抗性种群,6个种群为发展中抗性种群;在噁唑酰草胺 120 g/hm2有效成分剂量选择压下,4个千金子种群标记为抗性种群,5个种群为发展中抗性种群;在精噁唑禾草灵62.1 g/hm2有效成分剂量选择压下,6个千金子种群标记为抗性种群,2个种群为发展中抗性种群。对15个抗性千金子种群靶标酶基因片段的测序发现,9个种群共发生了4种ACCase基因突变类型,分别为ACCase基因 1999 位点色氨酸 (TGG) 突变为丝氨酸 (TCG)、1999位点色氨酸 (TGG) 突变为半胱氨酸 (TGT)、2027位点色氨酸 (TGG) 突变为丝氨酸 (TCG) 以及2027位点色氨酸 (TGG) 突变为半胱氨酸 (TGT/TGC)。其中,5个抗性千金子种群ACCase 基因突变频率均大于60%。研究表明,ACCase抑制剂类除草剂抗性千金子在上海市部分地区发生已较为严重,ACCase 基因突变是导致不同千金子种群对该类除草剂产生抗性的重要原因之一。     

几种除草剂对胡麻田裸燕麦(莜麦)、皮燕麦的防除效果

本文开展了唑啉草酯等7种除草剂对胡麻田裸燕麦、皮燕麦的防除试验研究,结果表明,在胡麻株高5~7cm,裸燕麦、皮燕麦3~5叶期施药,50g/L唑啉草酯乳油、108g/L高效氟吡甲禾灵乳油、15%精吡氟禾草灵乳油、240g/L烯草酮乳油和10%精喹禾灵乳油有效成分剂量分别为90、145.8、315、432和90g/hm2时,药后45d对裸燕麦的株防效达95.39%以上,鲜重防效达97.21%以上,胡麻增产率在82.79%以上;50g/L唑啉草酯乳油、108g/L高效氟吡甲禾灵乳油、15%精吡氟禾草灵乳油、15%炔草酯可湿性粉剂、10%精喹禾灵乳油和12.5%烯禾啶乳油有效成分剂量分别为90、145.8、270、135、90和412.5g/hm2时,药后45d对皮燕麦的株防效在94.66%以上,鲜重防效达96.13%以上,胡麻增产率在80.25%以上,各药在上述剂量下防治裸燕麦和皮燕麦后均对胡麻表现安全,可大面积示范推广。     

The response of ACCase-resistant Phalaris paradoxa populations involves two different target site mutations

Phalaris paradoxa (awned canary-grass) is an aggressive annual winter weed in wheat and other arable crops that is controlled mainly by ACCase-inhibiting herbicides: cyclohexanediones (DIMs), aryloxyphenoxypropionates (FOPs) and phenylpyrazolines (DENs, e.g. pinoxaden). The selection pressure imposed on the weed populations by repeated use of these herbicides has resulted in the evolution of increased numbers of ACCase-resistant populations of P. paradoxa in Israel and other countries. Two populations, Revadim (RV) and Mishmar Ha'emek (MH) that were exposed to differing weed and crop management tactics were investigated. Both populations were highly resistant to all FOPs, pinoxaden and cycloxydim, but responded differently to some DIMs. RV plants exhibited much higher resistance to tralkoxydim than MH plants, while showing similar low levels of resistance to tepraloxydim and clethodim. Both populations were equally susceptible to graminicides with other modes of action. The mutations responsible for the observed resistance were identified using PCR-RFLP and by sequencing the carboxyl transferase domain of the chloroplastic ACCase gene. RV plants possess a substitution of Asp₂₀₇₈ to Gly, whereas in MH population a mixture of Ile₂₀₄₁ to Asn or Asp₂₀₇₈ to Gly was found. Our study demonstrates that lack of herbicide and crop rotation may result in the evolution of diverse target site mutations and differential response of the whole plant to ACCase inhibitors.     

Multiple herbicide‐resistant Lolium rigidum (annual ryegrass) now dominates across the Western Australian grain belt

Lolium rigidum (annual or rigid ryegrass) is a widespread annual weed in cropping systems of southern Australia, and herbicide resistance in L. rigidum is a common problem in this region. In 2010, a random survey was conducted across the grain belt of Western Australia to determine the frequency of herbicide‐resistant L. rigidum populations and to compare this with the results of previous surveys in 1998 and 2003. During the survey, 466 cropping fields were visited, with a total of 362 L. rigidum populations collected. Screening of these populations with the herbicides commonly used for control of L. rigidum revealed that resistance to the ACCase‐ and ALS‐inhibiting herbicides was common, with 96% of populations having plants resistant to the ACCase herbicide diclofop‐methyl and 98% having plants resistant to the ALS herbicide sulfometuron. Resistance to another ACCase herbicide, clethodim, is increasing, with 65% of populations now containing resistant plants. Resistance to other herbicide modes of action was significantly lower, with 27% of populations containing plants with resistance to the pre‐emergent herbicide trifluralin, and glyphosate, atrazine and paraquat providing good control of most of the populations screened in this survey. Ninety five per cent of L. rigidum populations contained plants with resistance to at least two herbicide modes of action. These results demonstrate that resistance levels have increased dramatically for the ACCase‐ and ALS‐inhibiting herbicides since the last survey in 2003 (>95% vs. 70–90%); therefore, the use of a wide range of integrated weed management options are required to sustain these cropping systems in the future.     

Characterisation of target-site resistance to ACCase-inhibiting herbicides in the weed Alopecurus myosuroides (black-grass)

Resistance to aryloxyphenoxypropionate (AOPP), cyclohexanedione (CHD) and phenylurea herbicides was determined in UK populations of Alopecurus myosuroides Huds. Two populations (Oxford AA1, Notts. A1) were highly resistant (Resistance indices 13-->1000) to the AOPP and CHD herbicides fenoxaprop, diclofop, fluazifop-P and sethoxydim, but only marginally resistant to the phenylurea, chlorotoluron. Analyses of acetyl coenzyme A carboxylase (ACCase) activity showed that an insensitive ACCase conferred resistance to all the AOPP/CHD herbicides investigated. Another population, Oxford S1, showed no resistance to sethoxydim at the population level, but contained a small proportion of plants (<10%) with an insensitive ACCase. Genetic studies on the Notts A1 and Oxford S1 populations demonstrated that target site resistance conferred by an insensitive ACCase is monogenic, nuclearly inherited with the resistant allele showing complete dominance. Investigations of the molecular basis of resistance in the Notts A1 population showed that sethoxydim resistance in A myosuroides was associated with the substitution of an isoleucine in susceptible with a leucine in resistant plants, which has also been found in three other resistant grass-weed species (Setaria viridis (L) Beauv, Avena fatua L, Lolium rigidum Gaud).     

Multiple herbicide resistance in littleseed canarygrass (Phalaris minor): A threat to wheat production in India

Littleseed canarygrass (Phalaris minor Retz.), a troublesome weed of wheat in India, has evolved multiple herbicide resistance across three modes of action: photosynthesis at the photosystem II site A, acetyl‐coA carboxylase (ACCase), and acetolactate synthase inhibition. The multiple herbicide‐resistant (MHR) populations had a low level of sulfosulfuron resistance but a high level of resistance to clodinafop and fenoxaprop (ACCase inhibitors). Some of the populations had GR₅₀ (50% growth reduction) values for clodinafop that were 11.7‐fold greater than that of the most susceptible population. The clodinafop‐resistant populations also showed a higher level of cross‐resistance to fenoxaprop (fop group) but a low level of cross‐resistance to pinoxaden (den group). Although clodinafop and pinoxaden are from two different chemical families (fop and den groups), their same site of action is responsible for cross‐resistance behavior. The populations that were resistant to four groups of herbicides (phenylureas, sulfonylurea, aryloxyphenoxypropionate, and phenylpyrazolin) were susceptible to the triazine (metribuzin and terbutryn) and dinitroaniline (pendimethalin) herbicides. The P. minor populations that were resistant to the aryloxyphenoxypropionate and phenylurea herbicides were effectively controlled by the sulfonylurea herbicide, sulfosulfuron. In the fields infested with P. minor that was resistant to clodinafop, a sulfosulfuron application (25 g ha^?1) increased the wheat yield by 99.2% over that achieved using the recommended rate of clodinafop (60 g ha^?1). However, the evolution of multiple resistance against the four groups is a threat to wheat production. To prevent the spread of MHR P. minor populations, as well as the extension of multiple resistance to new chemicals, concerted efforts in developing and implementing a sound, integrated weed management program are needed. The integrated approach, consisting of crop and herbicide rotation with cultural and mechanical weed control tactics, should be considered as a long‐term resistance management strategy that will help to sustain wheat productivity and farmers' income.     

'Universal' primers for PCR-sequencing of grass chloroplastic acetyl-CoA carboxylase domains involved in resistance to herbicides

Primers were designed to amplify two regions involved in sensitivity to herbicides inhibiting the plastidic acetyl-CoA carboxylase (ACCase) from grasses (Poaceae). The first primer pair amplified a 551-bp amplicon containing a variable Ile/Leu codon at position 1781 in Alopecurus myosuroides sequence. The second primer pair amplified a 406-bp amplicon containing four variable codons (Trp/Cys, Ile/Asn, Asp/Gly, Gly/Ala) at positions 2027, 2041, 2078 and 2096, respectively, in A. myosuroides sequence. Both primer pairs amplified the targeted fragments from genes encoding plastidic ACCases, but not from the very similar genes encoding cytosolic ACCases. Clear DNA sequences were obtained from fresh or dried plant material from the field, and from 29 various grass species. Sequences revealed that the gene encoding plastidic ACCase in Poa annua and Festuca rubra contained a Leu₁₇₈₁ codon, in agreement with both species being inherently tolerant to herbicides inhibiting ACCase. Sequencing confirmed the hybrid origin of P. annua. Compared with ACCase enzyme assay, polymerase chain reaction is faster, can be performed from a single plant and suppresses the need for radioactive experiments. It can be completed with basic molecular biology laboratory equipment. It is the tool of choice for diagnosing resistance caused by alteration(s) of the plastidic ACCase.