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
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摘要
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。
The objectives of this study were to illuminate the sensitivity of Eleusine indica population in rice field of direct seeding to acetyl-CoA carboxylase(ACCase)-inhibiting herbicides and reveal its molecular resistance mechanism. Whole-plant bioassay showed that the population SJ-1 was highly resistant to metamifop, cyhalofop-butyl, fenoxaprop-p-ethyl, haloxyfop-R-methyl and sethoxydim, with resistance indexes of 56.6, 62.5, 128, 52.0 and 16.3, respectively. And it was slightly resistant to clethodim, with a resistance index of 4.86. By comparing the partial ACCase sequences derived from the resistant and susceptible populations, the SJ-1 population was found to contain one amino acid substitution of aspartate(GAT) for glycine(GGT) at the codon position 2078. This mutation in the ACCase gene might be responsible for the resistance to ACCase-inhibiting herbicides in the resistant population of E. indica.
The objectives of this study were to illuminate the sensitivity of Eleusine indica population in rice field of direct seeding to acetyl-CoA carboxylase(ACCase)-inhibiting herbicides and reveal its molecular resistance mechanism. Whole-plant bioassay showed that the population SJ-1 was highly resistant to metamifop, cyhalofop-butyl, fenoxaprop-p-ethyl, haloxyfop-R-methyl and sethoxydim, with resistance indexes of 56.6, 62.5, 128, 52.0 and 16.3, respectively. And it was slightly resistant to clethodim, with a resistance index of 4.86. By comparing the partial ACCase sequences derived from the resistant and susceptible populations, the SJ-1 population was found to contain one amino acid substitution of aspartate(GAT) for glycine(GGT) at the codon position 2078. This mutation in the ACCase gene might be responsible for the resistance to ACCase-inhibiting herbicides in the resistant population of E. indica.
引文
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[16]宗涛,李洁,刘祥英,等.湖南省部分地区棉田牛筋草(Eleusine indica)对精喹禾灵的抗性[J].植物保护,2015,41(2):58-63.ZONG T,LI J,LIU X Y,et al.Resistance of Eleusine indica to quizalofop-p-ethyl in some cotton fields in Hunan Province[J].Plant Prot,2015,41(2):58-63.
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[18]毕亚玲,王曹阳,谷刚,等.双环磺草酮除草活性及对水稻的安全性研究[J].农药学学报,2018,20(1):18-24.BI Y L,WANG C Y,GU G,et al.Herbicidal activity evaluation of benzobicyclon and its safety to rice[J].Chin J Pestic Sci,2018,20(1):18-24.
[19]ZHANG X Q,DEVINE M D.A possible point mutation of plastidic ACCase gene conferring resistance to sethoxydim in green foxtail(Setaria viridis)[C].Weed Sci Soc Am Abstr,40,2000:33.
[20]陈涛,张善磊,赵凌,等.ALS抑制剂类除草剂抗性水稻功能标记的开发与验证[J].中国水稻科学,2018,32(2):137-145.CHEN T,ZHANG S L,ZHAO L,et al.Development and verification of a functional marker associated with resistance to ALS inhibitor herbicide[J].Chin J Rice Sci,2018,32(2):137-145.
[21]YU L P,KIM Y S,TONG L.Mechanism for the inhibition of the carboxyltransferase domain of acetyl-coenzyme A carboxylase by pinoxaden[J].Proc Natl Acad Sci USA,2010,107(51):22072-22077.
[22]郭水良,赵铁桥.除草剂对杂草微观进化及多样性的影响[J].生物多样性,1997,5(4):301-306.GUO S L,ZHAO T Q.Influences of herbicide application on weed microevolution and diversity[J].Biodiv Science,1997,5(4):301-306.
[23]YU Q,AHMAD-HAMDANI M S,HAN H,et al.Herbicide resistance-endowing ACCase gene mutations in hexaploid wild oat(Avena fatua):insights into resistance evolution in a hexaploid species[J].Heredity(Edinb),2013,110(3):220-231.
[2]魏守辉,朱文达,杨小红,等.湖北省水稻田杂草的种类组成及其群落特征[J].华中农业大学学报,2013,32(2):44-49.WEI S H,ZHU W D,YANG X H,et al.Species composition and characterization of weed community in rice fields in Hubei Province[J].J Huazhong Agric Univ,2013,32(2):44-49.
[3]NIKOLAU B J,OHLROGGE J B,WURTELE E S.Plant biotincontaining carboxylases[J].Arch Biochem Biophys,2003,414(2):211-222.
[4]DéLYE C.Weed resistance to acetyl coenzyme A carboxylase inhibitors:an update[J].Weed Sci,2005,53(5):728-746.
[5]李蓉荣,袁国徽,郭文磊,等.八种除草剂对小麦田三种抗精唑禾草灵杂草的生物活性[J].植物保护学报,2016,43(3):507-513.LI R R,YUAN G H,GUO W L,et al.Biological activities of eight herbicides to three species of fenoxaprop-P-resistant weeds in wheat fields[J].Acta Phytophylacica Sinica,2016,43(3):507-513.
[6]黄世霞,王庆亚,张守栋.油菜田看麦娘对高效盖草能产生抗药性的现状研究[J].安徽农业科学,2006,34(9):1913-1914.HUANG S X,WANG Q Y,ZHANG S D.Study on the resistance Alopecurus aequalis sobol to haloxyfop-R-methyl[J].J Anhui Agric Sci,2006,34(9):1913-1914.
[7]DéLYE C,JASIENIUK M,LE CORRE V.Deciphering the evolution of herbicide resistance in weeds[J].Trends Genet,2013,29(11):649-658.
[8]夏向东,马洪菊,许孟涵,等.杂草对芳氧苯氧丙酸类(APPs)除草剂的抗性分子机理研究进展[J].农药学学报,2013,15(6):609-614.XIA X D,MA H J,XU M H,et al.Research progress on molecular mechanism of resistant weeds to aryloxyphenoxypropionate herbicides[J].Chin J Pestic Sci,2013,15(6):609-614.
[9]BECKIE H J,TARDIF F J.Herbicide cross resistance in weeds[J].Crop Prot,2012,35:15-28.
[10]CHA T S,NAJIHAH M G,SAHID I B,et al.Molecular basis for resistance to ACCase-inhibiting fluazifop in Eleusine indica from Malaysia[J].Pestic Biochem Physiol,2014,111:7-13.
[11]MCCULLOUGH P E,YU J L,CZARNOTA M A,et al.Physiological basis for metamifop selectivity on bermudagrass(Cynodon dactylon)and goosegrass(Eleusine indica)in cool-season turfgrasses[J].Weed Sci,2016,64(1):12-24.
[12]袁国徽,王恒智,赵宁,等.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[J].农药学学报,2016,18(3):304-310.YUAN G H,WANG H Z,ZHAO N,et al.Resistance to acetyl-CoAcarboxylase-inhibiting herbicides in Pseudosclerochloa kengiana and its molecular resistance mechanism[J].Chin J Pestic Sci,2016,18(3):304-310.
[13]OSUNA M D,GOULART I C G R,VIDAL R A,et al.Resistance to ACCase inhibitors in Eleusine indica from Brazil involves a target site mutation[J].Planta Daninha,2012,30(3):675-681.
[14]文马强,周小毛,刘佳,等.直播水稻田千金子对氰氟草酯抗性测定及抗性生化机理研究[J].南方农业学报,2017,48(4):647-652.WEN M Q,ZHOU X M,LIU J,et al.Resistance of Leptochloa chinensis(L)Nees.to cyhalofop-butyl in rice field of direct seeding and resistance biochemical mechanism research[J].J Southern Agric,2017,48(4):647-652.
[15]左平春,臧晓霞,陈仕红,等.稻稗对噁唑酰草胺的非靶标抗药性和对氰氟草酯的交互抗药性[J].杂草学报,2016,34(4):28-32.ZUO P C,ZANG X X,CHEN S H,et al.Non-target resistance to metamifop and cross-resistance to cyhalofop-butyl in Echinochloa phyllopogon(Stapf)Koss[J].Weed Sci,2016,34(4):28-32.
[16]宗涛,李洁,刘祥英,等.湖南省部分地区棉田牛筋草(Eleusine indica)对精喹禾灵的抗性[J].植物保护,2015,41(2):58-63.ZONG T,LI J,LIU X Y,et al.Resistance of Eleusine indica to quizalofop-p-ethyl in some cotton fields in Hunan Province[J].Plant Prot,2015,41(2):58-63.
[17]NORSWORTHY J K,WARD S M,SHAW D R,et al.Reducing the risks of herbicide resistance:best management practices and recommendations[J].Weed Sci,2012,60(SP1):31-62.
[18]毕亚玲,王曹阳,谷刚,等.双环磺草酮除草活性及对水稻的安全性研究[J].农药学学报,2018,20(1):18-24.BI Y L,WANG C Y,GU G,et al.Herbicidal activity evaluation of benzobicyclon and its safety to rice[J].Chin J Pestic Sci,2018,20(1):18-24.
[19]ZHANG X Q,DEVINE M D.A possible point mutation of plastidic ACCase gene conferring resistance to sethoxydim in green foxtail(Setaria viridis)[C].Weed Sci Soc Am Abstr,40,2000:33.
[20]陈涛,张善磊,赵凌,等.ALS抑制剂类除草剂抗性水稻功能标记的开发与验证[J].中国水稻科学,2018,32(2):137-145.CHEN T,ZHANG S L,ZHAO L,et al.Development and verification of a functional marker associated with resistance to ALS inhibitor herbicide[J].Chin J Rice Sci,2018,32(2):137-145.
[21]YU L P,KIM Y S,TONG L.Mechanism for the inhibition of the carboxyltransferase domain of acetyl-coenzyme A carboxylase by pinoxaden[J].Proc Natl Acad Sci USA,2010,107(51):22072-22077.
[22]郭水良,赵铁桥.除草剂对杂草微观进化及多样性的影响[J].生物多样性,1997,5(4):301-306.GUO S L,ZHAO T Q.Influences of herbicide application on weed microevolution and diversity[J].Biodiv Science,1997,5(4):301-306.
[23]YU Q,AHMAD-HAMDANI M S,HAN H,et al.Herbicide resistance-endowing ACCase gene mutations in hexaploid wild oat(Avena fatua):insights into resistance evolution in a hexaploid species[J].Heredity(Edinb),2013,110(3):220-231.