安徽省部分地区冬小麦田日本看麦娘Alopecurus japonicus对精鰁唑禾草灵的抗性发生现状及ACCase基因突变研究
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摘要
日本看麦娘Alopecurus japonicus是中国冬小麦田和油菜田主要恶性禾本科杂草之一。为了明确安徽省部分地区日本看麦娘对精齅唑禾草灵抗性发生情况及可能存在的抗性机制,本研究在安徽省天长市日本看麦娘发生严重区域冬小麦田共采集10个种群,采用温室盆栽法在整株水平上测定了不同种群对精齅唑禾草灵的抗性水平,扩增并比对了抗性和敏感种群之间靶标酶乙酰辅酶A羧化酶(Acetyl-CoA carboxylase,ACCase)基因部分序列的差异。结果显示,与敏感种群相比, 1 0个抗性种群对精齅唑禾草灵均产生了高水平抗性,抗性指数在30.50～58.55之间。不同抗性种群均发生了ACCase基因突变,其中8个种群发生了第1 781位异亮氨酸(Ile)到亮氨酸(Leu)突变,2个种群发生了第2 027位色氨酸(Trp)到半胱氨酸(Cys)突变。此外,各种群均具有较高的ACCase基因突变频率(≥80%)。研究表明,抗性日本看麦娘在安徽省部分地区发生较为严重,ACCase基因突变是导致不同日本看麦娘种群对精齅唑禾草灵产生抗性的重要原因之一。相对于第2 027位,日本看麦娘ACCase基因更倾向于在第1 781位产生突变以表现靶标抗性。
Japanese foxtail(Alopecurus japonicus) is a malignant grass weed mainly infesting winter wheat-and canola-growing fields in China. To determine the resistance level and the potential resistance mechanism of A. japonicus to fenoxaprop-p-ethyl, 10 populations of A. japonicus were collected from the wheat fields in Tianchang City, Anhui Province. Whole-plant dose-response experiments were performed to investigate the resistance 1evels of different populations to fenoxapropp-ethyl in the greenhouse. Partial sequences of acetyl-CoA carboxylase(ACCase) genes were subsequently amplified, sequenced, and compared between the resistant and susceptible plants. The results revealed that all 10 populations displayed high-level resistance to fenoxaprop-p-ethyl, and the resistance indices ranged from 30.50 to 58.55. Gene sequencing showed that the mutations of isoleucine(Ile) to leucine(Leu) at 1 781 position and tryptophan(Trp) to cysteine(Cys) at 2027 position existed in the ACCase genes of different populations, with eight of them had mutations at 1 781 position. Besides,all populations had mutant frequencies higher than 80%. Our results indicated that herbicide resistance has evolved to a severe stage in Anhui Province, considering that all 10 populations collected from different sites showed high-level resistance to fenoxaprop-p-ethyl. The Ile-1 781-Leu and Trp-2 027-Cys mutations in ACCase genes were very likely to be the key reasons which caused the fenoxaprop-pethyl resistance of A. japonicus populations. In addition, compared with 2 027 position, ACCase seems more prone to produce a mutation at 1 781 position in A. japonicus.
Japanese foxtail(Alopecurus japonicus) is a malignant grass weed mainly infesting winter wheat-and canola-growing fields in China. To determine the resistance level and the potential resistance mechanism of A. japonicus to fenoxaprop-p-ethyl, 10 populations of A. japonicus were collected from the wheat fields in Tianchang City, Anhui Province. Whole-plant dose-response experiments were performed to investigate the resistance 1evels of different populations to fenoxapropp-ethyl in the greenhouse. Partial sequences of acetyl-CoA carboxylase(ACCase) genes were subsequently amplified, sequenced, and compared between the resistant and susceptible plants. The results revealed that all 10 populations displayed high-level resistance to fenoxaprop-p-ethyl, and the resistance indices ranged from 30.50 to 58.55. Gene sequencing showed that the mutations of isoleucine(Ile) to leucine(Leu) at 1 781 position and tryptophan(Trp) to cysteine(Cys) at 2027 position existed in the ACCase genes of different populations, with eight of them had mutations at 1 781 position. Besides,all populations had mutant frequencies higher than 80%. Our results indicated that herbicide resistance has evolved to a severe stage in Anhui Province, considering that all 10 populations collected from different sites showed high-level resistance to fenoxaprop-p-ethyl. The Ile-1 781-Leu and Trp-2 027-Cys mutations in ACCase genes were very likely to be the key reasons which caused the fenoxaprop-pethyl resistance of A. japonicus populations. In addition, compared with 2 027 position, ACCase seems more prone to produce a mutation at 1 781 position in A. japonicus.
引文
[1]XU H L,ZHU X D,WANG H C,et al.Mechanism of resistance to fenoxaprop in Japanese foxtail(Alopecurus japonicus)from China[J].Pestic Biochem Physiol,2013,107(1):25-31.
[2]WU X B,ZHANG T,PAN L,et al.Germination requirements differ between fenoxaprop-p-ethyl resistant and susceptible Japanese foxtail(Alopecurus japonicus)biotypes[J].Weed Sci,2016,64(4):653-663.
[3]HERBERT D,WALKER K A,PRICE L J,et al.Acetyl-CoAcarboxylase-a graminicide target site[J].Pest Manag Sci,1997,50(1):67-71.
[4]HEAP I M.The international survey of herbicide resistant weeds[DB/OL].[2018-10-17].http://www.weedscience.org.
[5]POWLES S B,YU Q.Evolution in action:plants resistant to herbicides[J].Annu Rev Plant Biol,2010,61:317-347.
[6]KAUNDUN S S.Resistance to acetyl-CoA carboxylase-inhibiting herbicides[J].Pest Manag Sci,2014,70(9):1405-1417.
[7]MOHAMED I A,LI R Z,YOU Z G,et al.Japanese foxtail(Alopecurus japonicus)resistance to fenoxaprop and pinoxaden in China[J].Weed Sci,2012,60(2):167-171.
[8]TANG H W,LI J,DONG L Y,et al.Molecular bases for resistance to acetyl-coenzyme A carboxylase inhibitor in Japanese foxtail(Alopecurus japonicus)[J].Pest Manag Sci,2012,68(9):1241-1247.
[9]YUAN J S,TRANEL P J,STEWART C N JR.Non-target-site herbicide resistance:a family business[J].Trends Plant Sci,2007,12(1):6-13.
[10]GHANIZADEH H,HARRINGTON K C.Non-target site mechanisms of resistance to herbicides[J].Crit Rev Plant Sci,2017,36(1):24-34.
[11]毕亚玲,吴翠霞,郭文磊,等.抗精唑禾草灵的日本看麦娘ACCase基因突变[J].植物保护学报,2015,42(3):447-452.BI Y L,WU C X,GUO W L,et al.Mutation in the acetyl-CoAcarboxylase confers resistance to fenoxaprop-p-ethyl in Alopecurus japonicus populations[J].J Plant Prot,2015,42(3):447-452.
[12]BI Y L,LIU W T,GUO W L,et al.Molecular basis of multiple resistance to ACCase-and ALS-inhibiting herbicides in Alopecurus japonicus from China[J].Pestic Biochem Physiol,2016,126:22-27.
[13]赵宁,郭文磊,李伟,等.抗精唑禾草灵和甲基二磺隆看麦娘ACCase和ALS基因突变[J].植物保护学报,2017,44(5):833-840.ZHAO N,GUO W L,LI W,et al.Mutations in the acetyl-CoAcarboxylase and acetolactate synthase confer resistance to fenoxaprop-P-ethyl and mesosulfuron-methyl in Alopecurus aequalis populations[J].J Plant Prot,2017,44(5):833-840.
[14]SEEFELDT S S,JENSEN J E,FUERST E P.Log-logistic analysis of herbicide dose-response relationships[J].Weed Technol,1995,9(2):218-227.
[15]BECKIE H J,TARDIF F J.Herbicide cross resistance in weeds[J].Crop Prot,2012,35:15-28.
[16]POREBSKI S,BAILEY L G,BAUM B R.Modification of a CTABDNA extraction protocol for plants containing high polysaccharide and polyphenol components[J].Plant Mol Biol Rep,1997,15(1):8-15.
[17]ZHAO N,YAN Y Y,GE L A,et al.Target site mutations and cytochrome P450s confer resistance to fenoxaprop-p-ethyl and mesosulfuron-methyl in Alopecurus aequalis[J].Pest Manag Sci,2019,75(1):204-214.
[18]XU H L,ZHANG W P,ZHANG T,et al.Determination of ploidy level and isolation of genes encoding acetyl-CoA carboxylase in Japanese foxtail(Alopecurus japonicus)[J].PLoS One,2014,9(12):e114712.
[19]PETIT C,BAY G,PERNIN F,et al.Prevalence of cross-or multiple resistance to the acetyl-Coenzyme A carboxylase inhibitors fenoxaprop,clodinafop and pinoxaden in black-grass(Alopecurus myosuroides Huds.)in France[J].Pest Manag Sci,2010,66(2):168-177.
[20]YU Q,AHMAD-HAMDANI M S,HAN H P,et al.Herbicide resistance-endowing ACCase gene mutations in hexaploid wild oat(Avena fatua):insights into resistance evolution in a hexaploid species[J].Heredity,2013,110(3):220-231.
[21]ZHANG X Q,POWLES S B.Six amino acid substitutions in the carboxyl-transferase domain of the plastidic acetyl-CoA carboxylase gene are linked with resistance to herbicides in a Lolium rigidum population[J].New Phytol,2006,172(4):636-645.
[22]DéLYE C,MATéJICEK A,MICHEL S.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[J].Pest Manag Sci,2008,64(11):1179-1186.
[23]VILA-AIUB M M,YU Q,HAN H P,et al.Effect of herbicide resistance endowing Ile-1781-Leu and Asp-2078-Gly ACCase gene mutations on ACCase kinetics and growth traits in Lolium rigidum[J].J Exp Bot,2015,66(15):4711-4718.
[24]DU L,LIU W T,YUAN G H,et al.Cross-resistance patterns to ACCase-inhibitors in American sloughgrass(Beckmannia syzigachne Steud.)homozygous for specific ACCase mutations[J].Pestic Biochem Physiol,2016,126:42-48.
[25]HAWKINS N J,BASS C,DIXON A,et al.The evolutionary origins of pesticide resistance[J].Biol Rev,2018.
[26]NEVE P,VILA-AIUB M,ROUX F.Evolutionary-thinking in agricultural weed management[J].New Phytol,2009,184(4):783-793.
[27]ZHAO N,LI W,BAI S,et al.Transcriptome profiling to identify genes involved in mesosulfuron-methyl resistance in Alopecurus aequalis[J].Front Plant Sci,2017,8:1391.
[28]HAN H P,YU Q,CAWTHRAY G R,et al.Enhanced herbicide metabolism induced by 2,4-D in herbicide susceptible Lolium rigidum provides protection against diclofop-methyl[J].Pest Manag Sci,2013,69(9):996-1000.
[29]XU H L,LI J,ZHANG D,et al.Mutations at codon position 1999 of acetyl-CoA carboxylase confer resistance to ACCase-inhibiting herbicides in Japanese foxtail(Alopecurus japonicus)[J].Pest Manag Sci,2014,70(12):1894-1901.
[30]WANG H Z,HUANG Y Z,ZHANG L L,et al.Japanese foxtail(Alopecurus japonicus)management in wheat in China:seed germination,seedling emergence,and response to herbicide treatments[J].Weed Technol,2018,32(2):211-220.
[31]ZHAO N,LI Q,GUO W L,et al.Effect of environmental factors on germination and emergence of shortawn foxtail(Alopecurus aequalis)[J].Weed Sci,2018,66(1):47-56.
[2]WU X B,ZHANG T,PAN L,et al.Germination requirements differ between fenoxaprop-p-ethyl resistant and susceptible Japanese foxtail(Alopecurus japonicus)biotypes[J].Weed Sci,2016,64(4):653-663.
[3]HERBERT D,WALKER K A,PRICE L J,et al.Acetyl-CoAcarboxylase-a graminicide target site[J].Pest Manag Sci,1997,50(1):67-71.
[4]HEAP I M.The international survey of herbicide resistant weeds[DB/OL].[2018-10-17].http://www.weedscience.org.
[5]POWLES S B,YU Q.Evolution in action:plants resistant to herbicides[J].Annu Rev Plant Biol,2010,61:317-347.
[6]KAUNDUN S S.Resistance to acetyl-CoA carboxylase-inhibiting herbicides[J].Pest Manag Sci,2014,70(9):1405-1417.
[7]MOHAMED I A,LI R Z,YOU Z G,et al.Japanese foxtail(Alopecurus japonicus)resistance to fenoxaprop and pinoxaden in China[J].Weed Sci,2012,60(2):167-171.
[8]TANG H W,LI J,DONG L Y,et al.Molecular bases for resistance to acetyl-coenzyme A carboxylase inhibitor in Japanese foxtail(Alopecurus japonicus)[J].Pest Manag Sci,2012,68(9):1241-1247.
[9]YUAN J S,TRANEL P J,STEWART C N JR.Non-target-site herbicide resistance:a family business[J].Trends Plant Sci,2007,12(1):6-13.
[10]GHANIZADEH H,HARRINGTON K C.Non-target site mechanisms of resistance to herbicides[J].Crit Rev Plant Sci,2017,36(1):24-34.
[11]毕亚玲,吴翠霞,郭文磊,等.抗精唑禾草灵的日本看麦娘ACCase基因突变[J].植物保护学报,2015,42(3):447-452.BI Y L,WU C X,GUO W L,et al.Mutation in the acetyl-CoAcarboxylase confers resistance to fenoxaprop-p-ethyl in Alopecurus japonicus populations[J].J Plant Prot,2015,42(3):447-452.
[12]BI Y L,LIU W T,GUO W L,et al.Molecular basis of multiple resistance to ACCase-and ALS-inhibiting herbicides in Alopecurus japonicus from China[J].Pestic Biochem Physiol,2016,126:22-27.
[13]赵宁,郭文磊,李伟,等.抗精唑禾草灵和甲基二磺隆看麦娘ACCase和ALS基因突变[J].植物保护学报,2017,44(5):833-840.ZHAO N,GUO W L,LI W,et al.Mutations in the acetyl-CoAcarboxylase and acetolactate synthase confer resistance to fenoxaprop-P-ethyl and mesosulfuron-methyl in Alopecurus aequalis populations[J].J Plant Prot,2017,44(5):833-840.
[14]SEEFELDT S S,JENSEN J E,FUERST E P.Log-logistic analysis of herbicide dose-response relationships[J].Weed Technol,1995,9(2):218-227.
[15]BECKIE H J,TARDIF F J.Herbicide cross resistance in weeds[J].Crop Prot,2012,35:15-28.
[16]POREBSKI S,BAILEY L G,BAUM B R.Modification of a CTABDNA extraction protocol for plants containing high polysaccharide and polyphenol components[J].Plant Mol Biol Rep,1997,15(1):8-15.
[17]ZHAO N,YAN Y Y,GE L A,et al.Target site mutations and cytochrome P450s confer resistance to fenoxaprop-p-ethyl and mesosulfuron-methyl in Alopecurus aequalis[J].Pest Manag Sci,2019,75(1):204-214.
[18]XU H L,ZHANG W P,ZHANG T,et al.Determination of ploidy level and isolation of genes encoding acetyl-CoA carboxylase in Japanese foxtail(Alopecurus japonicus)[J].PLoS One,2014,9(12):e114712.
[19]PETIT C,BAY G,PERNIN F,et al.Prevalence of cross-or multiple resistance to the acetyl-Coenzyme A carboxylase inhibitors fenoxaprop,clodinafop and pinoxaden in black-grass(Alopecurus myosuroides Huds.)in France[J].Pest Manag Sci,2010,66(2):168-177.
[20]YU Q,AHMAD-HAMDANI M S,HAN H P,et al.Herbicide resistance-endowing ACCase gene mutations in hexaploid wild oat(Avena fatua):insights into resistance evolution in a hexaploid species[J].Heredity,2013,110(3):220-231.
[21]ZHANG X Q,POWLES S B.Six amino acid substitutions in the carboxyl-transferase domain of the plastidic acetyl-CoA carboxylase gene are linked with resistance to herbicides in a Lolium rigidum population[J].New Phytol,2006,172(4):636-645.
[22]DéLYE C,MATéJICEK A,MICHEL S.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[J].Pest Manag Sci,2008,64(11):1179-1186.
[23]VILA-AIUB M M,YU Q,HAN H P,et al.Effect of herbicide resistance endowing Ile-1781-Leu and Asp-2078-Gly ACCase gene mutations on ACCase kinetics and growth traits in Lolium rigidum[J].J Exp Bot,2015,66(15):4711-4718.
[24]DU L,LIU W T,YUAN G H,et al.Cross-resistance patterns to ACCase-inhibitors in American sloughgrass(Beckmannia syzigachne Steud.)homozygous for specific ACCase mutations[J].Pestic Biochem Physiol,2016,126:42-48.
[25]HAWKINS N J,BASS C,DIXON A,et al.The evolutionary origins of pesticide resistance[J].Biol Rev,2018.
[26]NEVE P,VILA-AIUB M,ROUX F.Evolutionary-thinking in agricultural weed management[J].New Phytol,2009,184(4):783-793.
[27]ZHAO N,LI W,BAI S,et al.Transcriptome profiling to identify genes involved in mesosulfuron-methyl resistance in Alopecurus aequalis[J].Front Plant Sci,2017,8:1391.
[28]HAN H P,YU Q,CAWTHRAY G R,et al.Enhanced herbicide metabolism induced by 2,4-D in herbicide susceptible Lolium rigidum provides protection against diclofop-methyl[J].Pest Manag Sci,2013,69(9):996-1000.
[29]XU H L,LI J,ZHANG D,et al.Mutations at codon position 1999 of acetyl-CoA carboxylase confer resistance to ACCase-inhibiting herbicides in Japanese foxtail(Alopecurus japonicus)[J].Pest Manag Sci,2014,70(12):1894-1901.
[30]WANG H Z,HUANG Y Z,ZHANG L L,et al.Japanese foxtail(Alopecurus japonicus)management in wheat in China:seed germination,seedling emergence,and response to herbicide treatments[J].Weed Technol,2018,32(2):211-220.
[31]ZHAO N,LI Q,GUO W L,et al.Effect of environmental factors on germination and emergence of shortawn foxtail(Alopecurus aequalis)[J].Weed Sci,2018,66(1):47-56.