山西省番茄早疫病菌对苯醚甲环唑的敏感性及抗性突变体的适合度
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摘要
为了解山西省番茄早疫病菌Alternaria solani对苯醚甲环唑的抗性,采用菌丝生长速率法检测了该病菌对苯醚甲环唑的敏感性,分析了抗性水平和抗性指数;同时通过室内诱导获得了苯醚甲环唑抗性突变体,检测了突变体的抗性遗传稳定性、交互抗性和适合度。结果表明:分离自大同、忻州、长治、晋中和运城5市的101株菌株的EC50范围为0.005～3.315 mg/L,均值为0.428 mg/L,其中敏感菌株占97.03%,低抗菌株占2.97%;抗性指数范围为0.295～0.447,均值为0.343。10株中抗突变体抗性均不能稳定遗传、生长速率减慢、产孢量降低、孢子萌发能力减弱、致病力下降;但在药剂压力下,突变体的致病力强于敏感菌株。苯醚甲环唑与代森锰锌、多抗霉素、啶酰菌胺无交互抗性,而与啶菌噁唑存在交互抗性。因此,今后需加强番茄早疫病菌对苯醚甲环唑的田间抗性监测,在生产过程中轮用或混用不同作用机制的药剂以延缓该病菌对苯醚甲环唑抗性的发展。
To understand the resistant situation of early blight pathogen Alternaria solani to difenoconazole in Shanxi Province, the sensitivity to difenoconazole were tested, and the resistance level and the resistance index were analyzed. Also, the difenoconazole-resistant mutants, which were induced in the laboratory, were studied on the genetic stability of resistance, the cross-resistance and the fitness. The results showed that the EC50 values of 101 isolates from Datong, Xinzhou, Changzhi, Jinzhong, and Yuncheng cities ranged from 0.005 to 3.315 mg/L with the mean value of 0.428 mg/L. Among these isolates, 97.03% showed sensitive to difenoconazole, and 2.97% isolates had low resistance. The resistance index ranged from 0.295 to 0.447 with the mean value of 0.343. Ten mutants with medium resistance to difenoconazole were derived from induction in the laboratory, but the resistance couldn't be inherited steadily. And their mycelial growth, sporulation, spore germination, and pathogenicity were declined.The pathogenicity of mutants was better than that of sensitive isolates under the stress of difenoconazole. No cross-resistance was detected between difenoconazole and mancozeb, polyoxin, or boscalid,excluding pyrisoxazole. Therefore, the sensitivity of A. solani to difenoconazole should be monitored continuously in future, and the mixed or rotated application of fungicides with different mechanism should be considered to postpone the development of resistance.
To understand the resistant situation of early blight pathogen Alternaria solani to difenoconazole in Shanxi Province, the sensitivity to difenoconazole were tested, and the resistance level and the resistance index were analyzed. Also, the difenoconazole-resistant mutants, which were induced in the laboratory, were studied on the genetic stability of resistance, the cross-resistance and the fitness. The results showed that the EC50 values of 101 isolates from Datong, Xinzhou, Changzhi, Jinzhong, and Yuncheng cities ranged from 0.005 to 3.315 mg/L with the mean value of 0.428 mg/L. Among these isolates, 97.03% showed sensitive to difenoconazole, and 2.97% isolates had low resistance. The resistance index ranged from 0.295 to 0.447 with the mean value of 0.343. Ten mutants with medium resistance to difenoconazole were derived from induction in the laboratory, but the resistance couldn't be inherited steadily. And their mycelial growth, sporulation, spore germination, and pathogenicity were declined.The pathogenicity of mutants was better than that of sensitive isolates under the stress of difenoconazole. No cross-resistance was detected between difenoconazole and mancozeb, polyoxin, or boscalid,excluding pyrisoxazole. Therefore, the sensitivity of A. solani to difenoconazole should be monitored continuously in future, and the mixed or rotated application of fungicides with different mechanism should be considered to postpone the development of resistance.
引文
Carter HE,Fraaije BA,West JS,Kelly SL,Mehl A,Shaw MW,Cools HJ.2014.Alterations in the predicted regulatory and coding regions of the sterol 14α-demethylase gene(CYP51)confer decreased azole sensitivity in the oilseed rape pathogen Pyrenopeziza brassicae.Molecular Plant Pathology,15(5):513-522
Chaerani R,Voorrips RE.2006.Tomato early blight(Alternaria solani):the pathogen,genetics,and breeding for resistance.Journal of General Plant Pathology,72(6):335-347
Chen FP,Fan JR,Zhou T,Liu XL.2012.Baseline sensitivity of Monilinia fructicola from China to the DMI fungicide SYP-Z048 and analysis of DMI-resistant mutants.Plant Disease,96(3):416-422
Dong JG.2001.Agricultural plant pathology(north edition).Beijing:China Agriculture Press,pp.385(in Chinese)[董金皋.2001.农业植物病理学(北方本).北京:中国农业出版社,pp.385]
Dong Y,Chen YF,Zhang HP.2015.Progresses of the microbial control of tomato early blight.Chinese Agricultural Science Bulletin,31(17):111-115(in Chinese)[董艳,陈永福,张和平.2015.番茄早疫病害微生物防治研究进展.中国农学通报,31(17):111-115]
FRAC.2016.FRAC code list 2016:fungicides sorted by mode of action(including FRAC code numbering).http://www.frac.info/docs/default-source/publications/frac-code-list/frac-code-list-2016.pdf?sfvrsn=2
Hamada MS,Yin YN,Ma ZH.2011.Sensitivity to iprodione,difenoconazole and fludioxonil of Rhizoctonia cerealis isolates collected from wheat in China.Crop Protection,30(8):1028-1033
Han YC,Xiang FY,Zeng XG,Chen FY,Gu YC.2016.Sensitivity of Colletotrichum spp.from strawberry to difenoconazole in Hubei Province.Journal of Plant Protection,43(3):525-526(in Chinese)[韩永超,向发云,曾祥国,陈丰滢,顾玉成.2016.湖北省草莓炭疽病菌对苯醚甲环唑的敏感性测定.植物保护学报,43(3):525-526]
Hao BQ,Ma LP,Zhu JS,Wang J,Qin S,Wang X,Qiao XW.2013.Investigation on vegetables production quality safety in Shanxi and Hebei provinces.Journal of Shanxi Agricultural Science,41(10):1116-1118,1121(in Chinese)[郝变青,马利平,朱九生,王静,秦曙,王霞,乔雄梧.2013.山西、河北蔬菜生产质量安全调查.山西农业科学,41(10):1116-1118,1121]
Kretschmer M,Leroch M,Mosbach A,Walker AS,Fillinger S,Mernke D,Schoonbeek HJ,Pradier JM,Leroux P,de Waard MA,et al.2009.Fungicide-driven evolution and molecular basis of multidrug resistance in field populations of the grey mould fungus Botrytis cinerea.PLoS Pathogens,5(12):e1000696
Liu G.2016.Sensitivity to tebuconazole of Rhizoctonia cerealis in Henan Province in China was declined.Pesticide Market News,(28):55(in Chinese)[刘刚.2016.河南省小麦纹枯病菌出现对苯醚甲环唑敏感性下降群体.农药市场信息,(28):55]
Liu YH,Wang KY,Jiang XY,Yi MQ,Wang HX.2003.Resistance of Rhizoctonia cerealis to tebuconazole and the biological characters of tebuconazole-resistant strains.Journal of Plant Protection,30(4):423-428(in Chinese)[刘英华,王开运,姜兴印,仪美芹,王怀训.2003.禾谷丝核菌对戊唑醇的抗性及抗药性菌系生物学特性.植物保护学报,30(4):423-428]
Luo CX,Cox KD,Amiri A,Schnabel G.2008.Occurrence and detection of the DMI resistance-associated genetic element‘Mona’in Monilinia fructicola.Plant Disease,92(7):1099-1103
Ma B,Tredway LP.2013.Induced overexpression of cytochrome P450sterol 14α-demethylase gene(CYP51)correlates with sensitivity to demethylation inhibitors(DMIs)in Sclerotinia homoeocarpa.Pest Management Science,69(12):1369-1378
Mirkovi?B,Tanovi?B,Stevi?M,Hrusti?J,Mihajlovi?M,Deliba?i?G,Vuk?a P.2015.Toxicity of mancozeb,chlorothalonil,captan,fluopyram,boscalid,and difenoconazole to Didymella applanata isolates from Serbia.Journal of Environmental Science and Health Part B:Pesticides,Food Contaminants,and Agricultural Wastes,50(12):845-850
Mondino P,Casanova L,Celio A,Oscar B,Leoni C,Alaniz S.2015.Sensitivity of Venturia inaequalis to trifloxystrobin and difenoconazole in Uruguay.Journal of Phytopathology,163(1):1-10
Qin C,Li QH,Liu RY.2015.Development situation and countermeasures of vegetable industry in Shanxi.China Agricultural Technology Extension,31(3):3-5(in Chinese)[秦潮,李庆华,刘瑞宇.2015.山西蔬菜产业发展情况与对策建议.中国农技推广,31(3):3-5]
Ren L,Shi XJ,Yao Z,Han JC,Zhao XJ.2017.Baseline sensitivity of Alternaria alternata f.sp.mali to tebuconazole and fitness of resistant mutants.Acta Phytopathologica Sinica,47(3):380-388(in Chinese)[任璐,史晓晶,姚众,韩巨才,赵晓军.2017.苹果斑点落叶病菌对戊唑醇敏感基线建立及抗性突变体适合度.植物病理学报,47(3):380-388]
Ren L,Yu L,Han JC,Liu HP.2014.Induction and characteristics of Botrytis cinerea resistant mutants to boscalid.Journal of Plant Protection,41(1):87-92(in Chinese)[任璐,余玲,韩巨才,刘慧平.2014.抗啶酰菌胺蔬菜灰霉病菌突变体的诱导及其生物学性状.植物保护学报,41(1):87-92]
Shi XJ,Ren L,Song YQ,Han JC,Liu HP,Zhang YJ.2015.Sensitivity of Alternaria solani to boscalid and control of boscalid resistance with commonly-used fungicides in Shanxi,China.Australasian Plant Pathology,44(3):327-334
Su P,Zhou ZQ,Hou H,Wang L,Zhu JL.2010.Examination of sensitivity of Botryosphaeria berengeriana f.sp.piricola to tebuconazole.Journal of Fruit Science,27(1):69-76(in Chinese)[苏平,周增强,侯珲,王丽,朱建兰.2010.苹果轮纹病菌对戊唑醇的敏感性检测.果树学报,27(1):69-76]
Wang HQ,Tian JS,Yan QP,Yuan SK.2008.Comparison of the sensitivity of Alternaria solani which causing tomato late blight to seven fungicides and its baseline-sensitivity to difenoconazole.Agrochemicals,47(4):294-296(in Chinese)[王海强,田家顺,严清平,袁善奎.2008.番茄早疫病菌对7种杀菌剂的敏感性比较及其对苯醚甲环唑的敏感性基线建立.农药,47(4):294-296]
Wang XK,Guo BB,Gao YY,Mu W,Liu F.2017.The toxicity of six triazole fungicides to Cladosporium fulvum and their safety and field efficacy in the control of tomato leaf mold.Journal of Plant Protection,44(4):671-678(in Chinese)[王晓坤,郭贝贝,高杨杨,慕卫,刘峰.2017.六种三唑类杀菌剂对番茄叶霉病菌的毒力及其安全性和田间防效评价.植物保护学报,44(4):671-678]
Xu JQ,Diao XW,Li H,Yang X,Wang BB,Liu QT.2016.Sensitivity to difenoconazole and tebuconazole of Rhizoctonia cerealis in Henan Province in China.Chinese Journal of Pesticide Science,18(5):582-588(in Chinese)[徐建强,刁兴旺,李恒,杨霞,王蓓蓓,刘庆涛.2016.中国河南省小麦纹枯病菌对苯醚甲环唑及戊唑醇的敏感性.农药学学报,18(5):582-588]
Ye T,Ma ZQ,Bi QY,Niu FS,Han XY,Zhang XF,Wang WQ,Zhang LH.2012.Research advances on the resistance of plant pathogenic fungi to SBIs fungicides.Chinese Journal of Pesticide Science,14(1):1-16(in Chinese)[叶滔,马志强,毕秋艳,牛芳胜,韩秀英,张小风,王文桥,张利辉.2012.植物病原真菌对甾醇生物合成抑制剂类(SBIs)杀菌剂的抗药性研究进展.农药学学报,14(1):1-16]
Yun YZ,Yu FW,Wang N,Chen HG,Yin YN,Ma ZH.2012.Sensitivity to silthiofam,tebuconazole and difenoconazole of Gaeumannomyces graminis var.tritici isolates from China.Pest Management Science,68(8):1156-1163
Zhang SY,Li L,Chen XL,Qi QQ,Lou HZ,Zhang CQ.2017.Identification of brown spot on fruit of Torreya grandis and screening of fungicides.Journal of Plant Protection,44(5):817-825(in Chinese)[张书亚,李玲,陈秀龙,戚钱钱,楼焕泽,张传清.2017.香榧果实褐斑病病原菌鉴定及防治药剂筛选.植物保护学报,44(5):817-825]
Zhao JJ,Han XY,Zhang XF,Ma ZQ,Wang WQ.2010.The sensitivity of Botrytis cinerea to difenoconazole and cross-resistance against diverse fungicides.Chinese Agricultural Science Bulletin,26(22):282-286(in Chinese)[赵建江,韩秀英,张小风,马志强,王文桥.2010.灰葡萄孢(Botrytis cinerea)对苯醚甲环唑的敏感性及其对不同杀菌剂的交互抗药性.中国农学通报,26(22):282-286]
Zhao XJ,Ren L,Yin H,Zhou JB,Han JC,Luo Y.2013.Sensitivity of Pseudoperonospora cubensis to dimethomorph,metalaxyl and fosetyl-aluminium in Shanxi of China.Crop Protection,43(1):38-44
Chaerani R,Voorrips RE.2006.Tomato early blight(Alternaria solani):the pathogen,genetics,and breeding for resistance.Journal of General Plant Pathology,72(6):335-347
Chen FP,Fan JR,Zhou T,Liu XL.2012.Baseline sensitivity of Monilinia fructicola from China to the DMI fungicide SYP-Z048 and analysis of DMI-resistant mutants.Plant Disease,96(3):416-422
Dong JG.2001.Agricultural plant pathology(north edition).Beijing:China Agriculture Press,pp.385(in Chinese)[董金皋.2001.农业植物病理学(北方本).北京:中国农业出版社,pp.385]
Dong Y,Chen YF,Zhang HP.2015.Progresses of the microbial control of tomato early blight.Chinese Agricultural Science Bulletin,31(17):111-115(in Chinese)[董艳,陈永福,张和平.2015.番茄早疫病害微生物防治研究进展.中国农学通报,31(17):111-115]
FRAC.2016.FRAC code list 2016:fungicides sorted by mode of action(including FRAC code numbering).http://www.frac.info/docs/default-source/publications/frac-code-list/frac-code-list-2016.pdf?sfvrsn=2
Hamada MS,Yin YN,Ma ZH.2011.Sensitivity to iprodione,difenoconazole and fludioxonil of Rhizoctonia cerealis isolates collected from wheat in China.Crop Protection,30(8):1028-1033
Han YC,Xiang FY,Zeng XG,Chen FY,Gu YC.2016.Sensitivity of Colletotrichum spp.from strawberry to difenoconazole in Hubei Province.Journal of Plant Protection,43(3):525-526(in Chinese)[韩永超,向发云,曾祥国,陈丰滢,顾玉成.2016.湖北省草莓炭疽病菌对苯醚甲环唑的敏感性测定.植物保护学报,43(3):525-526]
Hao BQ,Ma LP,Zhu JS,Wang J,Qin S,Wang X,Qiao XW.2013.Investigation on vegetables production quality safety in Shanxi and Hebei provinces.Journal of Shanxi Agricultural Science,41(10):1116-1118,1121(in Chinese)[郝变青,马利平,朱九生,王静,秦曙,王霞,乔雄梧.2013.山西、河北蔬菜生产质量安全调查.山西农业科学,41(10):1116-1118,1121]
Kretschmer M,Leroch M,Mosbach A,Walker AS,Fillinger S,Mernke D,Schoonbeek HJ,Pradier JM,Leroux P,de Waard MA,et al.2009.Fungicide-driven evolution and molecular basis of multidrug resistance in field populations of the grey mould fungus Botrytis cinerea.PLoS Pathogens,5(12):e1000696
Liu G.2016.Sensitivity to tebuconazole of Rhizoctonia cerealis in Henan Province in China was declined.Pesticide Market News,(28):55(in Chinese)[刘刚.2016.河南省小麦纹枯病菌出现对苯醚甲环唑敏感性下降群体.农药市场信息,(28):55]
Liu YH,Wang KY,Jiang XY,Yi MQ,Wang HX.2003.Resistance of Rhizoctonia cerealis to tebuconazole and the biological characters of tebuconazole-resistant strains.Journal of Plant Protection,30(4):423-428(in Chinese)[刘英华,王开运,姜兴印,仪美芹,王怀训.2003.禾谷丝核菌对戊唑醇的抗性及抗药性菌系生物学特性.植物保护学报,30(4):423-428]
Luo CX,Cox KD,Amiri A,Schnabel G.2008.Occurrence and detection of the DMI resistance-associated genetic element‘Mona’in Monilinia fructicola.Plant Disease,92(7):1099-1103
Ma B,Tredway LP.2013.Induced overexpression of cytochrome P450sterol 14α-demethylase gene(CYP51)correlates with sensitivity to demethylation inhibitors(DMIs)in Sclerotinia homoeocarpa.Pest Management Science,69(12):1369-1378
Mirkovi?B,Tanovi?B,Stevi?M,Hrusti?J,Mihajlovi?M,Deliba?i?G,Vuk?a P.2015.Toxicity of mancozeb,chlorothalonil,captan,fluopyram,boscalid,and difenoconazole to Didymella applanata isolates from Serbia.Journal of Environmental Science and Health Part B:Pesticides,Food Contaminants,and Agricultural Wastes,50(12):845-850
Mondino P,Casanova L,Celio A,Oscar B,Leoni C,Alaniz S.2015.Sensitivity of Venturia inaequalis to trifloxystrobin and difenoconazole in Uruguay.Journal of Phytopathology,163(1):1-10
Qin C,Li QH,Liu RY.2015.Development situation and countermeasures of vegetable industry in Shanxi.China Agricultural Technology Extension,31(3):3-5(in Chinese)[秦潮,李庆华,刘瑞宇.2015.山西蔬菜产业发展情况与对策建议.中国农技推广,31(3):3-5]
Ren L,Shi XJ,Yao Z,Han JC,Zhao XJ.2017.Baseline sensitivity of Alternaria alternata f.sp.mali to tebuconazole and fitness of resistant mutants.Acta Phytopathologica Sinica,47(3):380-388(in Chinese)[任璐,史晓晶,姚众,韩巨才,赵晓军.2017.苹果斑点落叶病菌对戊唑醇敏感基线建立及抗性突变体适合度.植物病理学报,47(3):380-388]
Ren L,Yu L,Han JC,Liu HP.2014.Induction and characteristics of Botrytis cinerea resistant mutants to boscalid.Journal of Plant Protection,41(1):87-92(in Chinese)[任璐,余玲,韩巨才,刘慧平.2014.抗啶酰菌胺蔬菜灰霉病菌突变体的诱导及其生物学性状.植物保护学报,41(1):87-92]
Shi XJ,Ren L,Song YQ,Han JC,Liu HP,Zhang YJ.2015.Sensitivity of Alternaria solani to boscalid and control of boscalid resistance with commonly-used fungicides in Shanxi,China.Australasian Plant Pathology,44(3):327-334
Su P,Zhou ZQ,Hou H,Wang L,Zhu JL.2010.Examination of sensitivity of Botryosphaeria berengeriana f.sp.piricola to tebuconazole.Journal of Fruit Science,27(1):69-76(in Chinese)[苏平,周增强,侯珲,王丽,朱建兰.2010.苹果轮纹病菌对戊唑醇的敏感性检测.果树学报,27(1):69-76]
Wang HQ,Tian JS,Yan QP,Yuan SK.2008.Comparison of the sensitivity of Alternaria solani which causing tomato late blight to seven fungicides and its baseline-sensitivity to difenoconazole.Agrochemicals,47(4):294-296(in Chinese)[王海强,田家顺,严清平,袁善奎.2008.番茄早疫病菌对7种杀菌剂的敏感性比较及其对苯醚甲环唑的敏感性基线建立.农药,47(4):294-296]
Wang XK,Guo BB,Gao YY,Mu W,Liu F.2017.The toxicity of six triazole fungicides to Cladosporium fulvum and their safety and field efficacy in the control of tomato leaf mold.Journal of Plant Protection,44(4):671-678(in Chinese)[王晓坤,郭贝贝,高杨杨,慕卫,刘峰.2017.六种三唑类杀菌剂对番茄叶霉病菌的毒力及其安全性和田间防效评价.植物保护学报,44(4):671-678]
Xu JQ,Diao XW,Li H,Yang X,Wang BB,Liu QT.2016.Sensitivity to difenoconazole and tebuconazole of Rhizoctonia cerealis in Henan Province in China.Chinese Journal of Pesticide Science,18(5):582-588(in Chinese)[徐建强,刁兴旺,李恒,杨霞,王蓓蓓,刘庆涛.2016.中国河南省小麦纹枯病菌对苯醚甲环唑及戊唑醇的敏感性.农药学学报,18(5):582-588]
Ye T,Ma ZQ,Bi QY,Niu FS,Han XY,Zhang XF,Wang WQ,Zhang LH.2012.Research advances on the resistance of plant pathogenic fungi to SBIs fungicides.Chinese Journal of Pesticide Science,14(1):1-16(in Chinese)[叶滔,马志强,毕秋艳,牛芳胜,韩秀英,张小风,王文桥,张利辉.2012.植物病原真菌对甾醇生物合成抑制剂类(SBIs)杀菌剂的抗药性研究进展.农药学学报,14(1):1-16]
Yun YZ,Yu FW,Wang N,Chen HG,Yin YN,Ma ZH.2012.Sensitivity to silthiofam,tebuconazole and difenoconazole of Gaeumannomyces graminis var.tritici isolates from China.Pest Management Science,68(8):1156-1163
Zhang SY,Li L,Chen XL,Qi QQ,Lou HZ,Zhang CQ.2017.Identification of brown spot on fruit of Torreya grandis and screening of fungicides.Journal of Plant Protection,44(5):817-825(in Chinese)[张书亚,李玲,陈秀龙,戚钱钱,楼焕泽,张传清.2017.香榧果实褐斑病病原菌鉴定及防治药剂筛选.植物保护学报,44(5):817-825]
Zhao JJ,Han XY,Zhang XF,Ma ZQ,Wang WQ.2010.The sensitivity of Botrytis cinerea to difenoconazole and cross-resistance against diverse fungicides.Chinese Agricultural Science Bulletin,26(22):282-286(in Chinese)[赵建江,韩秀英,张小风,马志强,王文桥.2010.灰葡萄孢(Botrytis cinerea)对苯醚甲环唑的敏感性及其对不同杀菌剂的交互抗药性.中国农学通报,26(22):282-286]
Zhao XJ,Ren L,Yin H,Zhou JB,Han JC,Luo Y.2013.Sensitivity of Pseudoperonospora cubensis to dimethomorph,metalaxyl and fosetyl-aluminium in Shanxi of China.Crop Protection,43(1):38-44