Cross-resistance patterns of winter wild oat (<Emphasis Type="Italic">Avena ludoviciana</Emphasis>) populations to ACCase inhibitor herbicides

The level of resistance and patterns of cross-resistance to clodinafop, sethoxydim, and pinoxaden were examined in 12 putative resistant and one susceptible populations of winter wild oat (Avena ludoviciana) collected from Fars Province, in the southwest of Iran. The responses of biomass and length of coleoptiles to the increasing dosages of the three herbicides were determined in both whole-plant and seed bioassays. In the whole-plant bioassay, all 12 putative resistant populations were found to be resistant to clodinafop with resistance ratios (R/S) ranging from 1.76 to >47.04. Most clodinafop-resistant populations exhibited low levels of cross-resistance to sethoxydim. Three highly sethoxydim-resistant populations, F2, S2, and ES4, were slightly resistant to clodinafop. Six populations (M1, M2, F2, S2, S4, and ES4) showed high cross-resistance to pinoxaden with R/S values as large as 10.73 to 40.29. A highly clodinafop-resistant population, M2, was more sensitive to pinoxaden than the susceptible population. The results of the seed bioassay resembled those obtained from the whole-plant experiment suggesting seed bioassay as an inexpensive, rapid method for screening-resistant genotypes.     

Cross-resistance to imidazolinone herbicides in chlorsulfuron-resistant Raphanus raphanistrum

Raphanus raphanistrum L has evolved widespread resistance to sulfonylureas in the Western Australia (WA) wheat belt. With the introduction of imidazolinone-tolerant (IT) wheat (Tritcum aestivum L) and IT canola (Brassica napus L) in the WA wheat belt, it is important to understand the status of cross-resistance in this weed to sulfonylurea and imidazolinone (Imi) herbicides. A study was conducted to examine cross-resistance between chlorsulfuron and Imi herbicides (a mixture of imazapic and imazapyr) in 46 R raphanistrum populations collected from across the WA wheat belt. Plants were treated with herbicides and assessed for phytotoxicity under glasshouse conditions. Of the 46 R raphanistrum populations, 32 were resistant to chlorsulfuron and four were resistant to imazapic + imazapyr. Of the 70% chlorsulfuron-resistant populations, 13% showed cross-resistance to imazapic + imazapyr. However, the cross-resistant populations treated with imazapic + imazapyr showed a lower resistance level than the chlorsulfuron-treated populations. These results suggest that weed populations with such cross-resistance will not be controlled effectively by Imi herbicides. Although the resistance levels of the cross-resistant populations to Imi herbicides were low, the cross-resistance levels of R raphanistrum should be determined before growing IT crops, particularly IT canola.     

Inhibition of acetolactate synthase in susceptible and resistant biotypes of Stellaria media

Acetolactate synthase (ALS) from one susceptible and two chlorsulfuronresistant biotypes of Stellaria media(L.) Vill. was assayed in the presence of eight known ALS inhibitors. As expected, ALS from the chlorsulfuronresistant biotypes (R1 and R2) showed reduced sensitivity to chlorsulfuron and other sulfonylurea herbicides. The patterns of cross-resistance varied, however, indicating that the alteration in ALS that confers chlorsulfuron resistance does not confer the same level of resistance to other sulfonylurea herbicides. The resistant biotypes were highly cross-resistant to sulfometuron-methyl and DPX-A7H81, but less cross-resistant to triasulfuron. Both R1 and R2 were highly cross-resistant to DTPS (N-[2,6-dichlorophenyl]-5,7-dimethyl-1,2,4-iriazolo[1,5a]pyrimidine-2-siilfoiiamide), but only slightly cross-resistant to imazamethahenz, an imidazolinone herbicide. The differences in the patterns of cross-resistance observed presumably reflect differences in the binding affinity of the herbicides for the altered ALS. The data presented suggest, but do not confirm, that R1 and R2 contain the same ALS mutation.     

Resistance of barnyardgrass (Echinochloa crus-galli) to atrazine and quinclorac

Two populations of Echinochloa crus-galli (R and I) exhibited resistance to quinclorac. Another population (X) exhibited resistance to quinclorac and atrazine. The R and I populations were collected from monocultures of rice in southern Spain. The X population was collected from maize fields subjected to the application of atrazine over several years. The susceptible (S) population of the same genus was collected from locations which had never been treated with herbicides. The quinclorac ED₅₀ value (dose causing 50% reduction in shoot fresh weight) for the R and I biotypes were 26- and 6-fold greater than for the S biotype. The X biotype was 10 times more tolerant to quinclorac than the S biotype and also showed cross-resistance to atrazine, being 82-fold more resistant to atrazine than the R, I and S biotypes. Chlorophyll fluorescence and Hill reaction analysis supported the view that the mechanism of resistance to atrazine in the X biotype was modification of the target site, the DI protein. Quinclorac at 20 mg litre^-1 did not inhibit photosynthetic electron transport in any of the test biotypes. The quinclorac I₅₀ values (herbicide dose needed for 50% Hill reaction reduction) of the S population was over 50000-fold higher than the atrazine I₅₀ value for the same S population, indicating that quinclorac is not a PS II inhibiting herbicide. Propanil at doses greater than 0·5 kg ha^-1 controlled all the biotypes. © 1997 SCI     

棉铃虫抗甲氧虫酰肼种群对12种杀虫剂的交互抗性

利用室内筛选获得的甲氧虫酰肼中等抗性种群(R, 30.57倍)和敏感种群(S),采用浸叶法测定了棉铃虫对12种常用杀虫剂的交互抗性。结果表明抗性种群对虫酰肼产生了13.57倍的中等水平交互抗性;对茚虫威的抗性倍数为3.05倍,无明显交互抗性;而对辛硫磷、毒死蜱、灭多威、高效氯氰菊酯、溴氰菊酯、虫螨腈、虱螨脲、氟啶脲、氟铃脲和甲氨基阿维菌素苯甲酸盐的抗性倍数在0.50 ~2.36倍之间,无交互抗性。试验结果提示,在棉铃虫对甲氧虫酰肼的抗药性治理中,轮换使用与甲氧虫酰肼没有交互抗性的杀虫剂将可有效延缓其抗药性发展。     

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     

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。     

Response of Arabidopsis thaliana to 22 ALS inhibitors: baseline toxicity and cross-resistance of csr1-1 and csr1-2 resistant mutants

The baseline toxicity of 22 acetolactate synthase (ALS)-inhibiting herbicides and the cross-resistance patterns of chlorsulfuron- and imazapyr-resistant (R) lines on these 22 ALS-inhibiting herbicides were investigated using the model species Arabidopsis thaliana. The 22 herbicides consisted of 18 sulfonylureas (SU), three imidazolinones (IMI) and one triazolopyrimidine (TP). The ED₅₀ values (doses of herbicides required to reduce dry matter by 50%) of the post-emergence-treated Col and Ler susceptible (S) lines ranged from 22 to 4822 mg ha⁻¹ and from 17 to 3143 mg ha⁻¹ respectively. The csr1-1 chlorsulfuron-resistant line (substitution of Pro₁₉₇ to Ser) conferred a high resistance to the only TP tested as well as to nine SU herbicides (R:S ratio ≥30), a low resistance to two SU herbicides (R:S≥5 and <30) and little or no resistance to the three IMI and seven other SU herbicides (R:S <5). This result contradicts the expectation that an ALS mutation selected by an SU herbicide confers high cross-resistance to other SU herbicides. We found that the efficacy of specific ALS inhibitors was different for different species and therefore could not be predicted from our results with A. thaliana; however, the cross-resistance patterns in A. thaliana were highly correlated with cross-resistance patterns in unrelated species with the same resistance mutation. These results have implications for resistance management.     

Resistance of Camelina microcarpa to acetolactate synthase inhibiting herbicides

An accession of Camelina microcarpa suspected to be resistant to sulfonylurea herbicides was identified in Oregon in 1998 field experiments. Greenhouse research confirmed that the putative resistant biotype was resistant to chlorsulfuron and metsulfuron on a whole plant level. Compared with the resistant (R) biotype, the susceptible (S) biotype was 1000 and 10 000‐fold more sensitive to metsulfuron and chlorsulfuron respectively. The R biotype was also resistant to other sulfonylurea, sulfonylaminocarbonyl‐triazolinone, imidazolinone and triazolopyrimidine herbicides. An in vivo enzyme assay indicated that acetolactate synthase (ALS) from the R plants required 111 times more chlorsulfuron to inhibit activity by 50% compared with the amount required to have a similar effect on ALS from S plants. Analysis of the nucleotide and amino acid sequences demonstrated that a single‐point mutation from G to T in the als1 gene conferred the change from the amino acid tryptophan to leucine at position 572 in the resistant biotype. This research confirmed that ALS inhibitor resistance in an Oregon accession of C. microcarpa is based on an altered target site conferred by a single‐point mutation.     

Biochemical mechanisms of cross-resistance to aryloxyphenoxypropionate and cyclohexanedione herbicides in populations of Avena spp.

Aryloxyphenoxypropionate (APP) and cyclohexanedione (CHD) herbicides are used extensively in the UK to control grass weeds, including Avena spp. (wild-oats). Reports of resistance to APP and CHD herbicides are a particular concern for the agricultural community. In this study, the responses of four UK Avena populations were characterized towards the APP herbicides fenoxaprop-P-ethyl and fluazifop-P-butyl, and towards the CHD herbicides cycloxydim and tralkoxydim. An A. sterilis ssp. ludoviciana population (T/41) was found to be highly resistant to fenoxaprop-P-ethyl and fluazifop-P-butyl, but did not show cross-resistance to cycloxydim and tralkoxydim. In contrast, one A. sterilis ssp. ludoviciana (T/11) and one A. fatua population (Dorset) showed partial resistance to both APP herbicides and also showed cross-resistance to the CHD herbicide tralkoxydim, but not to cycloxydim. Before this study, the biochemical mechanisms that confer resistance to the APP and CHD herbicides in UK Avena populations were unknown. Results from the present study show that an enhanced rate of metabolism of fenoxaprop-P-ethyl was found to confer resistance in the two partially resistant Avena populations (T/11 and Dorset), and the presence of an insensitive form of the target enzyme, ACCase, was responsible for target site resistance to fenoxaprop-P-ethyl and fluazifop-P-butyl in the highly resistant population T/41. Cross-resistance to the CHD herbicide tralkoxydim in the T/11 and Dorset populations was not conferred by insensitive ACCase, and was most probably caused by enhanced metabolism. This is the first report that resistance to fenoxaprop-P-ethyl can be conferred by enhanced metabolism in Avena spp.     

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

Background

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.

Results

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).

Conclusions

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.     

Sulfonylurea herbicide-resistant Monochoria vaginalis in Korean rice culture

Nine Monochoria vaginalis Pres1 accessions from Chonnam province, Korea were tested for resistance to the sulfonylurea herbicide, imazosulfuron, in whole-plant response bioassay. All accessions were confirmed resistant (R) to imazosulfuron. The GR50 (imazosulfuron concentration that reduced shoot dry weight by 50%) values of R accessions were 1112-3172 (accession #9) times higher than that of the standard susceptible (S) accession. Accession #9 exhibited cross-resistance to other sulfonylurea herbicides, bensulfuron-methyl, cyclosulfamuron and pyrazosulfuron-ethyl, but not to the imidazolinone herbicides, imazapyr and imazaquin. The R biotype could be controlled by other herbicides with different modes of action, such as mefenacet and pyrazolate, applied to soil at recommended rates. Foliar-applied herbicides, 2,4-D and bentazone, also controlled both the R and S biotypes. Sulfonylurea-based mixtures, except ethoxysulfuron plus fentrazamide, did not control resistant M. vaginalis. Rice yield was reduced 70% by resistant M. vaginalis that escaped pyrazosulfuron-ethyl plus molinate, compared with hand weeding in direct-seeded rice culture. In contrast, rice yield was reduced 44% by resistant M. vaginalis that survived the pyrazosulfuron-ethyl plus molinate treatment, compared with pyrazolate plus butachlor in transplanted rice culture. In vitro acetolactate synthase (ALS) activity of the R biotype was 183, 35, 130 and 31 times more resistant to imazosulfuron, bensulfuron-methyl, cyclosulfamuron and pyrazosulfuron-ethyl, respectively, than the S biotype. Imidazolinone herbicides, imazapyr and imazaquin had similar effect on in vitro ALS activity of the R and S biotypes. The in vivo ALS activity of the R biotype was also less affected than the S biotype by the sulfonylurea herbicides imazosulfuron and pyrazosulfuron-ethyl. Results of in vitro and in vivo ALS assays indicate that the resistance mechanism of M. vaginalis to sulfonylurea herbicides may be due, in part, to an alteration in the target enzyme, ALS. Since the level of resistance in the enzyme assay was much lower than that in the whole-plant assay, other mechanisms of resistance, such as herbicide metabolism, may be involved.     

Transformation a mutant Monochoria vaginalis acetolactate synthase (ALS) gene renders Arabidopsis thaliana resistant to ALS inhibitors

Acetolactate synthase (ALS) genes from Monochoria vaginalis resistant (R) and susceptible (S) biotypes against ALS inhibitors found in Korea revealed a single amino acid substitution of Proline (CCT), at 169th position based on the M. vaginalis ALS sequence numbering, to serine (TCT) in conserved domain A of the gene (equal to the proline 197 in Arabidopsis thaliana ALS gene sequence). A. thaliana plants transformed with the single mutated (Pro₁₆₉ to Ser) M. vaginalis ALS gene (including transit signal peptide) showed cross-resistance patterns to ALS-inhibiting herbicides, like as sulfonylurea-herbicide bensulfuron methyl (R/S factor of 9.5), imidazolinone-herbicide imazapyr (R/S factor of 5.1), and triazolopyrimidine-herbicide flumetsulam (R/S factor of 17.6) when measuring hypocotyls’ length of A. thaliana. The ALS activity from the transgenic A. thaliana plants confirmed the cross-resistance pattern to these herbicides like as R/S factor of 8.3 to bensulfuron methyl, 2.3 to imazapyr, and 13.2 to flumetsulam.     

Diclofop-methyl resistance in populations of Lolium spp. from central Italy

Populations of Lolium spp. collected in central Italy were screened for resistance to acetyl-coenzyme A carboxylase (ACCase)-inhibiting herbicides and compared with known susceptible and resistant Lolium rigidum (Gaud.) populations from Australia. Populations Roma'94 and Tuscania'97 were up to 8- and 7.5-fold more resistant to diclofop-methyl, respectively, than susceptible populations in pot experiments. However, populations Tarquinia'97 and Vetralla'94 were not resistant. Diclofop-methyl resistance levels in the Italian populations were lower than in the Australian populations SLR31 and WLR96 (16.5 and > 64 times more resistant than S respectively). In an agar germination test, Tuscania'97 showed low levels of cross-resistance to fluazifop-p-butyl, whereas no cross-resistance was found in the Roma'94 population.     

抗苄嘧磺隆雨久花乙酰乳酸合成酶突变的研究

本研究采用cDNA末端快速扩增技术(RACE)结合RT-PCR方法克隆抗苄嘧磺隆雨久花生物型和敏感性雨久花生物型乙酰乳酸合成酶(ALS)基因cDNA序列,并对测序结果进行比对分析。结果表明:与敏感性的雨久花ALS相比,公主岭(GZL)抗性生物型中第197位脯氨酸突变为组氨酸,第556位亮氨酸突变为苯丙氨酸;柳河(LH)抗性生物型中第358位天冬酰胺突变为天冬氨酸;磐石市(PS)抗性生物型中第525位缬氨酸突变为异亮氨酸。分析表明,高度保守区Domain A的第197位氨基酸残基的突变可能是导致公主岭稻区雨久花产生抗药性的主要原因之一,而其他抗性生物型抗性产生的原因有待进一步研究。     

斜纹夜蛾对茚虫威的抗药性汰选及交互抗性测定

为评估茚虫威抗性风险,在室内进行了斜纹夜蛾对茚虫威的抗性选育和交互抗性测定。经过10代6次室内抗性选育,获得了斜纹夜蛾对茚虫威抗性种群,与选育前相比,斜纹夜蛾对茚虫威的敏感性降低了15.63倍。抗性风险评估结果表明,斜纹夜蛾具有对茚虫威产生高水平抗性的风险。交互抗性测定发现,辛硫磷、高效氯氰菊酯和氟虫腈对茚虫威抗性种群的LC50值分别是同源对照种群的1.53、2.42和1.53倍,溴虫腈和灭多威对茚虫威抗性种群的LC50值分别是同源对照种群的0.78和0.96倍,表明茚虫威抗性种群对这几种杀虫剂未产生交互抗性。     

西瓜蔓枯病菌啶酰菌胺抗性突变体的生物学特性研究及其Sdh B基因位点检测

为评价西瓜蔓枯病菌对啶酰菌胺的抗性风险,了解其抗性机理,室内通过药剂驯化方法获得2株啶酰菌胺的抗性突变体XF21-3和YC60-1,测定了抗性突变体的生物学特性,并通过对Sdh B基因片段的测序比对,分析了西瓜蔓枯病菌对啶酰菌胺的抗性机理。生物测定结果表明:啶酰菌胺对2株抗性突变体的EC50值分别为108和124 μg/mL,抗性倍数（RR）分别为1 007和1 347,均为高抗菌株;抗性突变体的菌丝生长速率和产孢量均大于亲本菌株,但其致病性与亲本菌株无显著差异,对外界环境渗透压的敏感性低于亲本菌株;此外,啶酰菌胺与萎锈灵、戊唑醇、乙霉威及醚菌酯之间均不存在交互抗性,但与噻呋酰胺之间存在交互抗性。Sdh B基因片段测序及比对结果表明,高抗性突变体中Sdh B亚基277位上的氨基酸所对应的碱基由CAC突变为TAC,即由组氨酸（His）突变为酪氨酸（Tyr）。研究表明,西瓜蔓枯病菌在药剂选择压力下容易形成啶酰菌胺的抗性群体,且抗性突变体的离体适合度高于亲本菌株,此外,啶酰菌胺与同类型杀菌剂噻呋酰胺之间存在交互抗性,因此认为西瓜蔓枯病菌对啶酰菌胺具有中等抗性风险;同时进一步验证了Sdh B亚基277位上的氨基酸突变（His→Tyr,CAC→TAC）是西瓜蔓枯病菌对啶酰菌胺产生抗性的原因。     

Characterisation of Avena fatua populations with resistance to multiple herbicides

Avena fatua (wild oat) populations with resistance (R) to one or more herbicides have been described in numerous cropping systems worldwide. We previously reported that the R3 and R4 wild oat populations from Montana, USA, were resistant to four herbicides representing three different modes of action: tralkoxydim [acetyl‐CoA carboxylase (ACCase] inhibitor), imazamethabenz and flucarbazone [acetolactate synthase (ALS) inhibitors] and difenzoquat (growth inhibitor). We now quantify resistance levels of these populations to triallate [very long chain fatty acid (VLCFA) biosynthesis inhibitor], pinoxaden (ACCase inhibitor) and paraquat (photosystem I inhibitor). Glasshouse dose–response experiments showed that, compared with the means of two susceptible (S) populations, the R3 and R4 populations were 17.5‐ and 18.1‐fold more resistant to triallate, 3.6‐ and 3.7‐fold more resistant to pinoxaden, respectively, and 3.2‐fold (R3) more resistant to paraquat. Pre‐treatment of R plants with the cytochrome P450 inhibitor malathion partially reversed the resistance phenotype for flucarbazone (both populations), imazamethabenz (R4), difenzoquat (R4) and pinoxaden (R3), but not for tralkoxydim, fenoxaprop‐P‐ethyl or triallate. Target site point mutations known to confer resistance to ALS or ACCase inhibitors were not detected via DNA sequencing and allele‐specific PCR assays in R plants, suggesting the involvement of non‐target site resistance mechanism(s) for these herbicides. Together, our results complete the initial characterisation of wild oat populations that are resistant to seven (R3) or six (R4) herbicides from five or four mode of action families respectively.     

Characterization and diagnostic tests of resistance to acetohydroxyacid synthase inhibitors due to an Asp376Glu substitution in Amaranthus powellii

A population of Amaranthus powellii that had survived imazethapyr did not have any of the documented AHAS mutations causing resistance in this species in Ontario. The goal of this research was to confirm resistance in this population, determine its molecular basis and develop diagnostic tests. Whole plant dose-response experiments showed the resistant population had greater than 25-fold resistance to the selective agent imazethapyr at the whole plant level. Whole plant resistance to other herbicides ranged between 9-fold to flumetsulam and 85-fold to flucarbazone. This was confirmed at the enzyme level with resistance ranging between 25-fold to imazethapyr and 485-fold to thifensulfuron. AHAS from the resistant population had 16% higher specific activity than that of the susceptible population and it was also less sensitive to feedback inhibition by valine, leucine and isoleucine. Genomic DNA was extracted to PCR amplify and sequence the AHAS gene and to determine the applicability of diagnostic tests. Sequencing of the AHAS gene revealed a single point mutation of thymine₁₁₂₅ to guanine coding for an Asp₃₇₆Glu substitution. Two different diagnostic tests, restriction fragment length polymorphism and PCR amplification of specific allele, were able to correctly identify the resistant population from the susceptible control and from other resistant populations. Although this mutation appears to confer higher resistance to pyrithiobac and flucarbazone, two herbicides not registered in Eastern Canada, it endows plants with enough protection to allow survival to imazethapyr, the main selective agent in Ontario.     

Cross-resistance of Bidens subalternans to acetolactate synthase inhibitors in Brazil

Weeds resistant (R) to herbicides are widespread worldwide. Bidens subalternans is one of the most troublesome weeds in conventional soyabean fields in Brazil, and in a crop rotation system of cotton/soyabean and maize/soyabean some populations had evolved resistance to acetolactate synthase (ALS)-inhibiting herbicides. Bidens subalternans plants suspected of resistance were observed in soyabean fields where the main ALS-inhibiting herbicide sprayed is chlorimuron-ethyl. To confirm and characterise the resistance of B. subalternans to ALS inhibitors, whole-plant bioassays were conducted in 2006 and 2008. ALS in vivo enzyme bioassays were also conducted in 2007. In both bioassays, the R biotype showed cross-resistance to four chemical families of ALS-inhibiting herbicides. According to whole-plant level tests the R biotype showed 498-, 797-, 726- and >877-fold resistance to chlorimuron-ethyl, imazethapyr, cloransulam-methyl and pyrithiobac-sodium herbicides respectively. The R biotype was also 17-, 166-, 436- and 516-fold R to chlorimuron-ethyl, imazethapyr, cloransulam-methyl and pyrithiobac-sodium herbicides, respectively, based on the enzyme assay. Therefore, the herbicide-R B. subalternans can no longer be controlled by any ALS-inhibitor herbicides. Integrated control methods involving alternative herbicide with different modes of action are needed, to avoid yield losses in conventional soyabean fields in Brazil that are infested by ALS-R B. subalternans populations.