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乙酰乳酸合成酶抑制剂开发中的问题及对策 总被引:6,自引:0,他引:6
乙酰乳酸合成酶抑制剂开发中的问题及对策曹坳程(中国农业科学院植物保护研究所北京100094乙酰乳酸合成酶(ALS)抑制剂近年有了迅速发展,这类高效除草剂具有活性高,选择性强、对哺动物低毒等优异的特点,因而倍受关注,世界各大农药公司竞相投入巨额资金开发... 相似文献
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乙酰乳酸合成酶抑制剂的种类及其耐药性研究进展 总被引:2,自引:0,他引:2
乙酰乳酸合成酶(ALS)抑制剂在全世界已被迅速广泛认可和普遍使用。然而,由于ALS抑制剂的作用靶标单一,连续使用易诱发杂草产生抗药性等问题,ALS抑制剂类除草剂面临的杂草抗药性问题已越来越突出,并呈现出进一步恶化的趋势。本文概括了目前ALS抑制剂类除草剂的主要品种以及杂草对该类除草剂抗药性产生的主要机理,并针对因ALS基因突变导致的抗药性问题做了详细的阐述,为抗性基因应用的前景作了进一步展望。 相似文献
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植物乙酰乳酸合成酶抑制剂作用方式及机理研究进展 总被引:2,自引:2,他引:0
以拟南芥Arabidopsis thaliana等植物为主要对象,系统评述了乙酰乳酸合成酶(ALS)抑制剂在作用靶标、选择性机制、毒性机理及化学杀雄作用等方面的研究进展。ALS是磺酰脲类、咪唑啉酮类等多种除草剂的共同作用靶标,最新研究又发现苯磺隆、酰嘧磺隆等多种ALS抑制剂可作为敏感植物的化学杀雄剂。目前该研究领域的薄弱环节是ALS抑制剂的毒性机理,先后提出了支链氨基酸饥饿、核酸合成受阻、ALS底物积累、养分转运障碍、无氧呼吸等假说,但均未能被证实。借助高通量的代谢组学、转录组学、蛋白质组学检测技术将能够更全面地揭示ALS抑制剂的生理生化效应,为研究其毒性机理提供新证据。 相似文献
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乙酰乳酸合成酶及其抑制剂研究新进展 总被引:1,自引:0,他引:1
乙酰乳酸合成酶(AHAS)是支链氨基酸生物合成途径中的一个关键酶,是绿色除草剂的重要作用靶标。由于此生物合成过程只存在于植物和微生物体内,因此该类抑制剂对哺乳动物具有生物安全性。近年来,随着AHAS三维结构的阐明,人们不仅深入了解了已有抑制剂的作用机制,并且依此设计开发了一些新型的抑制剂, 拓展了其在抑菌活性方面的生物学功能。文章对近年来AHAS及其抑制剂的最新研究进展进行了综述,重点就AHAS的酶学特征、结构特征及结合方式,以AHAS为靶标的新颖除草活性化合物的设计开发以及AHAS抑制剂的抗菌生物活性研究进展等问题详细进行了总结,以期为设计开发靶向AHAS的新型除草剂或抗菌药物提供参考。 相似文献
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除草剂与乙酰乳酸合成酶相互作用的分子机理研究进展 总被引:5,自引:0,他引:5
综述了酵母乙酰乳酸合成酶(ScALS)与磺酰脲类除草剂氯嘧磺隆(chlorimuron-ethyl,CE)形成的复合物在0.28 nm分辨率下的晶体结构及拟南芥乙酰乳酸合成酶(AtALS)与磺酰脲类和咪唑啉酮类除草剂复合物的三维结构。除草剂的分子结构与酶、底物并不相似,但它们与酶形成的复合物可阻断底物进入酶活性位点通路而起抑制作用。连接磺酰脲的10个氨基酸残基同样连接咪唑喹啉酸,另有6个残基只与磺酰脲而不与咪唑喹啉酸相连,有2个残基只与咪唑喹啉酸而不与磺酰脲相连,即两种抑制剂占据了特别的重叠位点,但以不同方式连接。抗性杂草的产生是因为突变株ALS的残基位点变异,从而引起除草剂与ALS结合方式的变化。这些研究对进一步理解除草剂与靶分子的作用方式及除草剂的分子药物设计具有重要的指导作用。 相似文献
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氟乐灵抗药性杂草研究进展 总被引:5,自引:0,他引:5
氟乐灵抗药性杂草研究进展黄建中,孙丙耀,李扬汉(南京农业大学杂草研究室210095)Ran于1970年首次公开报道欧洲千里光(Seneciovulgaris)对三氮苯类除草剂西玛津和阿特拉津产生抗性以来,不断有杂草对除草剂产生抗性的报道。杂草抗药性问... 相似文献
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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。 相似文献
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近年来长江下游地区稻田耳叶水苋Ammannia arenaria H.B.K.危害十分严重。采用盆栽法首次测定了耳叶水苋对苄嘧磺隆等药剂的抗性水平,同时分析了其抗性和敏感种群间乙酰乳酸合成酶(ALS)基因的DNA序列及其RNA表达差异。结果表明:采自浙江嘉兴(JX110)、江苏苏州(JS039)、浙江宁波(NB0143-05)和安徽广德(AH014)的耳叶水苋生物型对苄嘧磺隆的抗性指数(RI)分别为67.90、17.59、44.63和8.37,对苄嘧磺隆表现出中高水平抗性的生物型对五氟磺草胺、双草醚及咪唑乙烟酸也产生了低水平的抗性。获得了耳叶水苋ALS基因全长核苷酸序列2235 bp,编码667个氨基酸,仅发现NB0143-05等3种抗性生物型ALS酶的氨基酸序列非保守区第93位的亮氨酸被脯氨酸取代。然而,NB0143-05的ALS酶对ALS抑制剂的敏感性大幅度降低(RI 37.04),且在苄嘧磺隆处理后4 d的ALS基因表达量是敏感生物型(HZ001)的1.86倍。这表明,ALS酶对药剂的敏感性降低以及被苄嘧磺隆诱导后ALS基因表达量显著增加,很可能是耳叶水苋生物型NB0143-05对ALS抑制剂产生抗性的原因。 相似文献
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Tatiana Vega Gabriela Breccia Graciela Nestares María Laura Mayor Roxana Zorzoli Liliana Picardi 《Pest management science》2009,65(9):991-995
BACKGROUND: Rapid and efficient diagnostic tests for early screening of herbicide resistance are convenient alternatives to field screening methods. There is a need for a quick, reliable and cost‐effective method for rapid diagnosis of imidazolinone resistance in sunflower (Helianthus annuus L.). RESULTS: Two seed germination bioassays were developed. Seeds from three sunflower inbred lines differing in resistance to imidazolinones were germinated either on solid culture medium or placed in plastic pots filled with commercial perlite. After 8 days incubation under controlled conditions, both assays successfully distinguished susceptible genotype from the resistant and intermediate ones. The susceptible genotype showed arrested root growth at all herbicide treatments (root length < 1 cm). The resistant genotype developed a complete root system even when exposed to the highest dose of herbicide. However, no definite differences were observed for the intermediate and resistant genotypes with respect to root growth under the different herbicide treatments. CONCLUSION: The simple and rapid screening assays described in the present study were useful in discriminating imidazolinone resistance at the seedling stage. Therefore, these bioassays could be potential tools for early screening of imidazolinone resistance genes from large sunflower populations. Copyright © 2009 Society of Chemical Industry 相似文献
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小麦田麦家公对苯磺隆的抗性机理 总被引:2,自引:0,他引:2
为明确麦田阔叶杂草麦家公Lithospermum arvense L.对苯磺隆的抗性机理,以苯磺隆抗性和敏感型麦家公为材料,比较分析这2个生物型麦家公靶标酶乙酰乳酸合成酶(acetolactate synthase,ALS)、解毒酶谷胱甘肽-S-转移酶(glutathione-S-transferase,GST)以及保护酶过氧化物酶(peroxidase,POD)、超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)对苯磺隆的响应差异性。结果表明,抗性麦家公ALS对苯磺隆的敏感性较敏感型麦家公显著下降,苯磺隆的抑制中浓度分别为0.187、0.036 μmol/L。苯磺隆胁迫后,抗性和敏感型麦家公ALS活性都出现下降,但抗性麦家公ALS活性可恢复,而敏感型麦家公ALS活性则不能恢复;2个生物型麦家公GST活性都能被苯磺隆诱导,但抗性麦家公GST累计活性为29.31 U,高于敏感型麦家公(25.90 U);抗性麦家公SOD累计活性为24.49 U,较敏感型麦家公(19.31 U)高,且具有较强的恢复能力;抗性麦家公POD和CAT累计活性分别为126.92~550.68 U和41.41~77.19 U,也高于敏感型麦家公的93.75~271.04 U、42.17~57.28 U。因此,靶标酶ALS对苯磺隆敏感性减弱是麦家公产生抗性的一个重要原因,解毒酶GST、SOD、POD和CAT活性升高可能与抗性有关。 相似文献
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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... 相似文献
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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 相似文献
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Summary A biotype of Conyza albida resistant to imazapyr was discovered on a farm in the province of Seville, Spain, on land that had been continuously treated with this herbicide. This is the first reported occurrence of target site resistance to acetolactate synthase (ALS)-inhibiting herbicides in C. albida . In order to characterize this resistant biotype, dose–response experiments, absorption and translocation assays, metabolism studies, ALS activity assays and control with alternative herbicides were performed. Dose–response experiments revealed a marked difference between resistant (R) and susceptible (S) biotypes with a resistance factor [ED50 (R)/ED50 (S)] of 300. Cross-resistance existed with amidosulfuron, imazethapyr and nicosulfuron. Control of both biotypes using alternative herbicides was good using chlorsulfuron, triasulfuron, diuron, simazine, glyphosate and glufosinate. The rest of the herbicides tested did not provide good control for either biotype. There were no differences in absorption and translocation between the two biotypes, the maximum absorption reached about 15%, and most of the radioactivity taken up remained in the treated leaf. The metabolism pattern was similar and revealed that both biotypes may form polar metabolites with similar retention time (Rf ). The effect of several ALS inhibitors on ALS (target site) activity measured in leaf extracts from both biotypes was investigated. Only with imazapyr and imazethapyr did the R biotype show a higher level of resistance than the S biotype [I50 (R)/I50 (S) value of 4.0 and 3.7 respectively]. These data suggest that the resistance to imazapyr found in the R biotype of C. albida results primarily from an altered target site. 相似文献
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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 相似文献