九种杀菌剂对新拟盘多毛孢病菌的室内毒力作用
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摘要
新拟盘多毛孢叶斑病是在草莓上发现的一种新病害。本研究对采自北京昌平、山东诸城、云南保山及广东惠州等地草莓Fragaria×ananassa发病叶片进行了组织分离,结合形态特征和ITS系统发育分析对所分离获得的8株疑似拟盘多毛孢叶斑病病菌进行了鉴定,并采用菌丝生长速率法测定了病原菌对8种代表性杀菌剂及新型药剂——双苯菌胺的敏感性。结果表明:所分离得到的病原菌为新拟盘多毛孢Neopestalotiopsis clavispora;双苯菌胺对8株新拟盘多毛孢N. clavispora的毒力最强,平均EC_(50)值为(0.004±0.002)μg/mL,可作为防治草莓叶斑病的首选药剂;其次为咪鲜胺、嘧菌酯、戊唑醇、氟环唑、苯醚甲环唑和腈菌唑,平均EC_(50)值在0.006～0.803μg/mL之间;百菌清毒力较低,而代森锰锌对新拟盘多毛孢N.clavispora几乎没有抑制作用。研究结果对合理使用杀菌剂防治草莓新病害——新拟盘多毛孢叶斑病提供参考。
Neopestalotiopsis leaf spot is a new disease on strawberry. The pathogens of the new strawberry leaf spot disease from Changping district, Beijing, Zhucheng city, Shandong Province,Baoshan city, Yunnan Province, Huizhou city, Guangdong Province and other regions were isolated and identified. The pathogen was identified by morphological analysis and ITS phylogenetic analysis.Sensitivities of 8 isolates to 8 commonly used fungicides and SYP-14288 were determined by indoor inoculation. The results showed that the pathogen was Neopestalotiopsis clavispora. SYP-14288 had a strong inhibitory effect on the mycelia growth of 8 isolates of N. clavispora, with an average EC_(50) value of(0.004 ± 0.002) μg/mL, which could be used as the first choice for the control of strawberry leaf spot disease. The mean of EC_(50) values were 0.006-0.803 μg/mL for prochloraz, azoxystrobin, tebuconazole,epoxiconazole, difenoconazole, and myclobutanil, respectively. Chlorothalonil exhibited lower toxicity.while mancozeb had the lowest inhibitory activity. Those results are important for the rational application of fungicides in the chemical control of Neopestalotiopsis leaf spot, a new disease of strawberry.
Neopestalotiopsis leaf spot is a new disease on strawberry. The pathogens of the new strawberry leaf spot disease from Changping district, Beijing, Zhucheng city, Shandong Province,Baoshan city, Yunnan Province, Huizhou city, Guangdong Province and other regions were isolated and identified. The pathogen was identified by morphological analysis and ITS phylogenetic analysis.Sensitivities of 8 isolates to 8 commonly used fungicides and SYP-14288 were determined by indoor inoculation. The results showed that the pathogen was Neopestalotiopsis clavispora. SYP-14288 had a strong inhibitory effect on the mycelia growth of 8 isolates of N. clavispora, with an average EC_(50) value of(0.004 ± 0.002) μg/mL, which could be used as the first choice for the control of strawberry leaf spot disease. The mean of EC_(50) values were 0.006-0.803 μg/mL for prochloraz, azoxystrobin, tebuconazole,epoxiconazole, difenoconazole, and myclobutanil, respectively. Chlorothalonil exhibited lower toxicity.while mancozeb had the lowest inhibitory activity. Those results are important for the rational application of fungicides in the chemical control of Neopestalotiopsis leaf spot, a new disease of strawberry.
引文
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[13]赵景楠,马喆,刘正坪,等.草莓拟盘多毛孢叶斑病的病原菌[J].菌物学报,2016,35(1):114-120.ZHAO J N,MA Z,LIU Z P,et al.Pestalotiopsis clavispora causing leaf spot on strawberry[J].Mycosystema,2016,35(1):114-120.
[14]赵宇,钱恒伟,徐鹏程,等.青岛市草莓根腐病病原菌分离及鉴定[J].中国植保导刊,2016,36(1):43-46,82.ZHAO Y,QIAN H W,XU P C,et al.Isolation and identification of pathogen of strawberry root rot in Qingdao[J].China Plant Prot,2016,36(1):43-46,82.
[15]VALENCIA A L,TORRES R,LATORRE B A.First report of Pestalotiopsis clavispora and Pestalotiopsis spp.causing postharvest stem end rot of avocado in Chile[J].Plant Dis,2011,95(4):492.
[16]LAZAROTTO M,MUNIZ M F B,POLETTO T,et al.First report of Pestalotiopsis clavispora causing leaf spot of Carya illinoensis in Brazil[J].Plant Dis,2012,96(12):1826.
[17]FENG Y R,LIU B S,SUN B B.First report of leaf blotch caused by Pestalotiopsis clavispora on Rosa chinensis in China[J].Plant Dis,2014,98(7):1009.
[18]YANG L N,MIAO X Y, BAI Q R,et al.NeoPestalotiopsis clavispora causing leaf spot on Phedimus aizoon var.latifolius,a new disease in China[J].Plant Dis,2017,101(11):1952-1953.
[19]CHEN Y J,ZENG L,SHU N,et al.Pestalotiopsis-like species causing gray blight disease on Camellia sinensis in China[J].Plant Dis,2018,102(1):98-106.
[20]徐英,马森,杨奎,等.40%戊唑醇悬浮剂对草莓炭疽病的毒力测定及田间防效[J].农药,2017,56(11):853-855.XU Y,MA S,YANG K,et al.The toxicity determination and field efficacy trials of tebuconazole 40%SC against Fragaria ananassa colletotrichum[J].Agrochemicals,2017,56(1 1):853-855.
[21]张佳,张璨,芦帆,等.草莓灰霉病菌对嘧菌酯的抗性检测及抗性菌株的生物学特性研究[J].植物病理学报,2016,46(1):124-130.ZHANG J,ZHANG C,LU F,et al.Detection of resistance to azoxystrobin and characterization of the azoxystrobin-resistant isolates in Botrytis cinerea from strawberry[J].Acta Phytopathol Sin,2016,46(1):124-130.
[22]张颂函,陈秀,赵莉,等.6种杀菌剂防治草莓白粉病的田间药效评价[J].农药科学与管理,2017,38(6):55-58.ZHANG S H,CHEN X,ZHAO L,et al.Evaluation on field efficacy of six fungicides against powdery mildew on strawberry[J].Pestic Sci Admin,2017,38(6):55-58.
[23]方中达.植病研究方法[M].北京:中国农业出版社,1998:427.FANG Z D.Plant disease research methods[M].Beijing:China Agriculture Press,1998:427.
[24]温浩,张桂军,毕扬,等.草莓棒孢叶斑病的病原菌鉴定及其对9种杀菌剂的敏感性[J].农药学学报,2019,21(3):279-284.WEN H,ZHANG G J,BI Y,et al.Pathogen identification of Corynespora leaf spot on strawberry and its sensitivity to ninefungicides[J].Chin J Pestic Sci,2019,21(3):279-284.
[25]宋玉.拟盘多毛孢属(Pestalotiopsis)真菌系统学研究及条形码基因的评价[D].贵阳:贵州大学,2015.SONG Y.Taxonomy of the genus Pestalotiopsis and evaluation of its DNA barcoding[D].Guiyang:Guizhou University,2015.
[26]WHITE T J,BRUNS T,LEE S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[M]//PCR Protocols.Elsevier,1990:315-322.DOI:10.1016/b978-0-12-3721 80-8.50042-1
[27]慕立义.植物化学保护研究方法[M].北京:中国农业出版社,1994:76-81.MU L Y.Research methods of chemical plant protection[M].Beijing:China Agriculture Press,1994:76-8 1.
[28]韦继光,徐同,潘秀湖,等.拟盘多毛孢属的分类学研究进展[J].广西农业生物科学,2006,25(1):78-85.WEI J G,XU T,PAN X H,et al.Progress of research on taxonomy of Pestalotiopsis[J].J Guangxi Agric Biol Sci,2006,25(1):78-85.
[29]JEEWON R,LIEW E C Y,HYDE K D.Phylogenetic evaluation of species nomenclature of Pestalotiopsis in relation to host association[J].Fungal Divers,2004,17:39-55.
[30]JEEWON R,LIEW E C Y,SIMPSON J A,et al.Phylogenetic significance of morphological characters in the taxonomy of Pestalotiopsis species[J].Mol Phylogenetics Evol,2003,27(3):372-383.
[31]韩永超,曾祥国,向发云,等.湖北省草莓炭疽病菌对咪鲜胺的敏感性[J].农药学学报,2014,16(5):535-540.HAN Y C,ZENG X G,XIANG F Y,et al.Sensitivity of Colletotrichum spp.from strawberry to prochloraz in Hubei Province of China[J].Chin J Pestic Sci,2014,16(5):535-540.
[32]耿文龙,张夏兰,胡盼,等.草莓褐色叶斑病病原菌生物学特性及其室内药剂筛选[C]//中国植物病理学会2012年学术年会论文集.北京:中国农业科学技术出版社,2012:28.GENG W L,ZHANG X L,HU P,et al.Biological characteristics and laboratory evaluation of fungitoxicants against Pilidium lythri causing tan-brown leaf spot of strawberry[C]//Proceedings of the Annual Meeting of Chinese Society for Plant Pathology(2012).Beijing:China Agricultural Science and Technology Press,2012:28.
[33]倪笑霞.双苯菌胺的抑菌谱及其对辣椒疫霉的作用机制研究[D].北京:中国农业大学,2013.NI X X.Study on the antifungal spectrum and the mode of action of the novel fungicide SYP-14288 to Phytophthora capsici[D].Beijing:China Agricultural University,2013.
[34]兰杰,陈宣明,司乃国,等.双苯菌胺对菌核病生物活性及田间应用技术研究[M]//中国植物病害化学防治研究:第9卷,2014:51-57.LAN J,CHEN X M,SI N G,et al.The lab and field efficacy of SYP14288 against cucumber downy mildew[M]//Chemical control of plant diseases in China:Vol.9,2014:51-57.
[2]LEE S,CROUS P W,WINGFIELD M J.Pestalotioid fungi from Restionaceae in the cape floral kingdom[J].Stud Mycol,2006,55:175-187.
[3]MAHARACHCHIKUMBURA S S N,HYDE K D,GROENEWALD J Z,et al. Pestalotiopsis revisited[J].Stud Mycol,2014,79:121-186.
[4]黄泽余,周志权,黄平明,等.广西红树林真菌病害调查初报[J].广西科学院学报,1997,13(4):41-45.HUANG Z Y,ZHOU Z Q,HUANG P M,et al.Brief report on fungus disease attached to mangrove in Guangxi[J].J Guangxi Acad Sci,1997,13(4):41-45.
[5]石凌波,李媛,费诺亚,等.蓝莓拟盘多毛孢叶斑病病原菌鉴定[J].中国南方果树,2017,46(1):24-28.SHI L B,LI Y,FEI N Y,et al.Identification of the Pestalotiopsis pathogen of blueberry leaf spot[J].South China Fruits,2017,46(1):24-28.
[6]赵洪海,岳清华,梁晨.蓝莓拟盘多毛孢枝枯病的病原菌[J].菌物学报,2014,33(3):577-583.ZHAO H H,YUE Q H,LIANG C.The pathogen causing Pestalotiopsis twig dieback of blueberry[J].Mycosystema,2014,33(3):577-583.
[7]ESPINOZA J G,BRICENO E X,KEITH L M,et al.Canker and twig dieback of blueberry caused by Pestalotiopsis spp.and a Truncatella spp.in Chile[J].Plant Dis,2008,92(10):1407-1414.
[8]TEIXEIRA M A,MARTINS R M S,VIEIRA R F,et al.In vitro identification and control of Pestalotiopsis longisetula fungus,pathogens strawberry crop[J].Revista Agrogeoambiental,2015,7(3):59-65.
[9]WEBER R W S,ENTROP A P.Dactylonectria tor resens is as the main component of the black root rot complex of strawberries andraspberries in northern Germany[J].Erwerbs-Obstbau,2017,59(3):157-169.
[10]OBREGON V G,MENEGUZZIN G,IBANEZ J M,et al.First report of Neopestalotiopsis clavispora causing root and crown rot on strawberry plants in Argentina[J].Plant Dis,2018,102(9):1856.
[11]MAHAPATRA S,BANERJEE J, KUMAR K,et al.Leaf spot and fruit rot of strawberry caused by Neopestalotiopsis clavispora in IndoGangetic plains of India[J].Indian Phytopathol,2018,71(2):279-283.
[12]CHAMORRO M,AGUADO A,de los SANTOS B.First report of root and crown rot caused by Pestalotiopsis clavispora(Neopestalotiopsis clavispora)on strawberry in Spain[J].Plant Dis,2016,100(7):1495.
[13]赵景楠,马喆,刘正坪,等.草莓拟盘多毛孢叶斑病的病原菌[J].菌物学报,2016,35(1):114-120.ZHAO J N,MA Z,LIU Z P,et al.Pestalotiopsis clavispora causing leaf spot on strawberry[J].Mycosystema,2016,35(1):114-120.
[14]赵宇,钱恒伟,徐鹏程,等.青岛市草莓根腐病病原菌分离及鉴定[J].中国植保导刊,2016,36(1):43-46,82.ZHAO Y,QIAN H W,XU P C,et al.Isolation and identification of pathogen of strawberry root rot in Qingdao[J].China Plant Prot,2016,36(1):43-46,82.
[15]VALENCIA A L,TORRES R,LATORRE B A.First report of Pestalotiopsis clavispora and Pestalotiopsis spp.causing postharvest stem end rot of avocado in Chile[J].Plant Dis,2011,95(4):492.
[16]LAZAROTTO M,MUNIZ M F B,POLETTO T,et al.First report of Pestalotiopsis clavispora causing leaf spot of Carya illinoensis in Brazil[J].Plant Dis,2012,96(12):1826.
[17]FENG Y R,LIU B S,SUN B B.First report of leaf blotch caused by Pestalotiopsis clavispora on Rosa chinensis in China[J].Plant Dis,2014,98(7):1009.
[18]YANG L N,MIAO X Y, BAI Q R,et al.NeoPestalotiopsis clavispora causing leaf spot on Phedimus aizoon var.latifolius,a new disease in China[J].Plant Dis,2017,101(11):1952-1953.
[19]CHEN Y J,ZENG L,SHU N,et al.Pestalotiopsis-like species causing gray blight disease on Camellia sinensis in China[J].Plant Dis,2018,102(1):98-106.
[20]徐英,马森,杨奎,等.40%戊唑醇悬浮剂对草莓炭疽病的毒力测定及田间防效[J].农药,2017,56(11):853-855.XU Y,MA S,YANG K,et al.The toxicity determination and field efficacy trials of tebuconazole 40%SC against Fragaria ananassa colletotrichum[J].Agrochemicals,2017,56(1 1):853-855.
[21]张佳,张璨,芦帆,等.草莓灰霉病菌对嘧菌酯的抗性检测及抗性菌株的生物学特性研究[J].植物病理学报,2016,46(1):124-130.ZHANG J,ZHANG C,LU F,et al.Detection of resistance to azoxystrobin and characterization of the azoxystrobin-resistant isolates in Botrytis cinerea from strawberry[J].Acta Phytopathol Sin,2016,46(1):124-130.
[22]张颂函,陈秀,赵莉,等.6种杀菌剂防治草莓白粉病的田间药效评价[J].农药科学与管理,2017,38(6):55-58.ZHANG S H,CHEN X,ZHAO L,et al.Evaluation on field efficacy of six fungicides against powdery mildew on strawberry[J].Pestic Sci Admin,2017,38(6):55-58.
[23]方中达.植病研究方法[M].北京:中国农业出版社,1998:427.FANG Z D.Plant disease research methods[M].Beijing:China Agriculture Press,1998:427.
[24]温浩,张桂军,毕扬,等.草莓棒孢叶斑病的病原菌鉴定及其对9种杀菌剂的敏感性[J].农药学学报,2019,21(3):279-284.WEN H,ZHANG G J,BI Y,et al.Pathogen identification of Corynespora leaf spot on strawberry and its sensitivity to ninefungicides[J].Chin J Pestic Sci,2019,21(3):279-284.
[25]宋玉.拟盘多毛孢属(Pestalotiopsis)真菌系统学研究及条形码基因的评价[D].贵阳:贵州大学,2015.SONG Y.Taxonomy of the genus Pestalotiopsis and evaluation of its DNA barcoding[D].Guiyang:Guizhou University,2015.
[26]WHITE T J,BRUNS T,LEE S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[M]//PCR Protocols.Elsevier,1990:315-322.DOI:10.1016/b978-0-12-3721 80-8.50042-1
[27]慕立义.植物化学保护研究方法[M].北京:中国农业出版社,1994:76-81.MU L Y.Research methods of chemical plant protection[M].Beijing:China Agriculture Press,1994:76-8 1.
[28]韦继光,徐同,潘秀湖,等.拟盘多毛孢属的分类学研究进展[J].广西农业生物科学,2006,25(1):78-85.WEI J G,XU T,PAN X H,et al.Progress of research on taxonomy of Pestalotiopsis[J].J Guangxi Agric Biol Sci,2006,25(1):78-85.
[29]JEEWON R,LIEW E C Y,HYDE K D.Phylogenetic evaluation of species nomenclature of Pestalotiopsis in relation to host association[J].Fungal Divers,2004,17:39-55.
[30]JEEWON R,LIEW E C Y,SIMPSON J A,et al.Phylogenetic significance of morphological characters in the taxonomy of Pestalotiopsis species[J].Mol Phylogenetics Evol,2003,27(3):372-383.
[31]韩永超,曾祥国,向发云,等.湖北省草莓炭疽病菌对咪鲜胺的敏感性[J].农药学学报,2014,16(5):535-540.HAN Y C,ZENG X G,XIANG F Y,et al.Sensitivity of Colletotrichum spp.from strawberry to prochloraz in Hubei Province of China[J].Chin J Pestic Sci,2014,16(5):535-540.
[32]耿文龙,张夏兰,胡盼,等.草莓褐色叶斑病病原菌生物学特性及其室内药剂筛选[C]//中国植物病理学会2012年学术年会论文集.北京:中国农业科学技术出版社,2012:28.GENG W L,ZHANG X L,HU P,et al.Biological characteristics and laboratory evaluation of fungitoxicants against Pilidium lythri causing tan-brown leaf spot of strawberry[C]//Proceedings of the Annual Meeting of Chinese Society for Plant Pathology(2012).Beijing:China Agricultural Science and Technology Press,2012:28.
[33]倪笑霞.双苯菌胺的抑菌谱及其对辣椒疫霉的作用机制研究[D].北京:中国农业大学,2013.NI X X.Study on the antifungal spectrum and the mode of action of the novel fungicide SYP-14288 to Phytophthora capsici[D].Beijing:China Agricultural University,2013.
[34]兰杰,陈宣明,司乃国,等.双苯菌胺对菌核病生物活性及田间应用技术研究[M]//中国植物病害化学防治研究:第9卷,2014:51-57.LAN J,CHEN X M,SI N G,et al.The lab and field efficacy of SYP14288 against cucumber downy mildew[M]//Chemical control of plant diseases in China:Vol.9,2014:51-57.