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
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摘要
对采自山东省诸城市草莓叶斑病发病叶片上的病原菌进行了分离及鉴定,通过室内人工接种法确定该病原菌的致病性,并采用菌丝生长速率法测定了其对双苯菌胺等9种杀菌剂的敏感性。结果表明:形态学结合ITS系统发育分析确定该病原菌为多主棒孢霉Corynespora cassiicola。室内毒力测定结果显示:9种供试杀菌剂中,双苯菌胺对C. cassiicola菌丝生长有很强的抑制作用,其平均EC50值为(0.27±0.18)μg/mL,可作为防治草莓叶斑病的首选药剂;其次为氟环唑、腈菌唑、咪鲜胺、苯醚甲环唑、嘧菌酯和戊唑醇,平均EC50值分别为(0.66±0.21)、(0.85±0.25)、(1.4±1.2)、(1.8±0.3)、(3.8±3.0)和(4.5±2.6)μg/mL;而该病原菌对百菌清与代森锰锌的敏感性最低。研究结果可为草莓棒孢叶斑病的化学防治提供参考。
The pathogen of a new strawberry leaf spot disease collected from Zhucheng city, Shandong Province was isolated and identified. Its pathogenicity was determined by indoor inoculation. Its susceptibility to 9 fungicides such as SYP-14288 was determined by mycelial growth rate method. The results showed that the pathogen was Corynespora cassiicola according to the morphological analysis and ITS phylogenetic analysis. The results also showed that SYP-14288 had a strong inhibitory effect on the mycelia growth of C. cassiicola, with an average EC50 value of(0.27 ± 0.18) μg/mL, which could be used as the first choice for the control of strawberry leaf spot disease. The mean of EC50 values were(0.66 ± 0.21) μg/mL,(0.85 ± 0.25) μg/mL,(1.4 ± 1.2) μg/mL,(1.8 ± 0.3) μg/mL,(3.8 ± 3.0) μg/mL and(4.5 ± 2.6) μg/mL for epoxiconazole, myclobutanil, prochloraz, difenoconazole, azoxystrobin and tebuconazole, respectively. Chlorothalonil and mancozeb had the lowest toxicity. The results have provided a reference for the control of Corynespora leaf spot on strawberry.
The pathogen of a new strawberry leaf spot disease collected from Zhucheng city, Shandong Province was isolated and identified. Its pathogenicity was determined by indoor inoculation. Its susceptibility to 9 fungicides such as SYP-14288 was determined by mycelial growth rate method. The results showed that the pathogen was Corynespora cassiicola according to the morphological analysis and ITS phylogenetic analysis. The results also showed that SYP-14288 had a strong inhibitory effect on the mycelia growth of C. cassiicola, with an average EC50 value of(0.27 ± 0.18) μg/mL, which could be used as the first choice for the control of strawberry leaf spot disease. The mean of EC50 values were(0.66 ± 0.21) μg/mL,(0.85 ± 0.25) μg/mL,(1.4 ± 1.2) μg/mL,(1.8 ± 0.3) μg/mL,(3.8 ± 3.0) μg/mL and(4.5 ± 2.6) μg/mL for epoxiconazole, myclobutanil, prochloraz, difenoconazole, azoxystrobin and tebuconazole, respectively. Chlorothalonil and mancozeb had the lowest toxicity. The results have provided a reference for the control of Corynespora leaf spot on strawberry.
引文
[1]ZHANG M, MA Q Z, ZHAI N P, et al. Occurrence of outbreak of leaf spot caused by Corynespora cassiicola in strawberry in China[J].Plant Dis, 2018, 102(10):2037.
[2]DIXON L J, SCHLUB R L, PERNEZNY K, et al. Host specialization and phylogenetic diversity of Corynespora cassiicola[J].Phytopathology, 2009, 99(9):1015-1027.
[3]李长松,张眉,李林,等.山东省黄瓜棒孢叶斑病(褐斑病)病原菌鉴定和防治[J].中国蔬菜, 2009, 1(18):29-33.LI C S, ZHANG M, LI L, et al. Identification of cucumber target leaf spot(brown spot)pathogen and its control[J]. China Vegetables,2009, 1(18):29-33.
[4]禾丽菲,李晓旭,朱佳美,等.不同杀菌剂对黄瓜靶斑病菌的毒力作用特性比较[J].农药学学报, 2018, 20(1):25-32.HE L F, LI X X, ZHU J M, et al. Comparison of toxicity properties of different types of fungicides against Corynespora cassiicola on cucumber[J]. Chin J Pestic Sci, 2018, 20(1):25-32.
[5]KWON M K, KANG B R, CHO B H, et al. Occurrence of target leaf spot disease caused by Corynespora cassicola on cucumber in Korea[J]. Plant Pathol, 2003, 52(3):424.
[6]蓝国兵,谭耀华,何自福,等.黄瓜褐斑病(Corynespora cassiicola)在广东首次报道[J].植物保护, 2012, 38(5):197-200.LAN G B, TAN Y H, HE Z F, et al. First report of Cucumis sativus brown spot disease infected by Corynespora cassiicola in Guangdong Province[J]. Plant Prot, 2012, 38(5):197-200.
[7]杜玉宁,查仙芳,杨金凤,等.宁夏温室黄瓜新病害靶斑病的药剂筛选[J].江苏农业科学, 2013, 41(1):132-133.DU Y N, ZHA X F, YANG J F, et al. Screening of Corynespora cassiicolaon cucumber in greenhouse of Ningxia[J]. Jiangsu Agricultural Sciences, 2013, 41(1):132-133.
[8]杨双娟,顾兴芳,张圣平,等.黄瓜棒孢叶斑病(Corynespora cassiicola)的研究概况[J].中国蔬菜, 2012, 1(4):1-9.YANG S J, GU X F, ZHANG S P, et al. Research progress on cucumber target leaf spot(Corynespora cassiicola)[J]. China Vegetables, 2012, 1(4):1-9.
[9]张乃楼,李亚美,康文强,等.辽宁省黄瓜靶斑病菌对苯醚甲环唑和戊唑醇的敏感性[J].农药学学报, 2014, 16(4):452-456.ZHANG N L, LI Y M, KANG W Q, et al. Sensitivity of Corynespora cassiicolato difenoconazole and tebuconazole in Liaoning Province[J]. Chin J Pestic Sci, 2014, 16(4):452-456.
[10]张中义,李继新,关国经,等.烤烟棒孢霉叶斑病病原菌鉴定[J].中国烟草学报, 2008, 14(6):44-47.ZHANG Z Y, LI J X, GUAN G J, et al. Identification of fungal pathogen of corynespora leaf spot in flue-cured tobacco[J]. Acta Tabacaria Sinica, 2008, 14(6):44-47.
[11]XUE L H, LIU Y, LI C J. First report of leaf spot caused by Corynespora cassiicola on Ixeris polycephala in China[J]. Plant Dis,2017, 101(8):1552.
[12]YEH Y H, KIRSCHNER R. First report of Corynespora leaf spot on Vitex rotundifolia caused by Corynespora cassiicola in Taiwan[J].Plant Dis, 2017, 101(8):1550.
[13]BAIYEE B, PORNSURIYA C, ITO S I, et al. Trichoderma spirale T76-1 displays biocontrol activity against leaf spot on lettuce(Lactuca sativa L.)caused by Corynespora cassiicola or Curvularia aeria[J]. Biol Control, 2019, 129:195-200.
[14]BUTLER S, YOUNG-KELLY H, RAPER T, et al. First report of target spot caused by Corynespora cassiicolaon cotton in Tennessee[J]. Plant Dis, 2016, 100(2):535.
[15]HUANG X, ZHENG X, XU J, et al. First report of brown leaf spot on honeysuckle caused by Corynespora cassiicola in China[J]. Plant Dis,2016, 100(11):2326.
[16]XU J, QI X, ZHENG X. First report of Corynespora leaf spot on sweet potato caused by Corynespora cassiicolain China[J].Australasian Plant Disease Notes, 2016, 100(10):2163.
[17]ONOFRE R B, MERTELY J C, AGUIAR F M, et al. First report of target spot caused by Corynespora cassiicola on blueberry in North America[J]. Plant Dis, 2016, 100(2):528-529.
[18]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.
[19]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.
[20]WHITE T J, BRUNS T, LEE S, et al. 38 amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[J]. Pcr Protoc, 1990:315-322.
[21]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, 1994:76-81.
[22]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.
[23]WANG Z W, NI X X, PENG Q, et al. The novel fungicide SYP-14288 acts as an uncoupler against Phytophthora capsici[J]. Pestic Biochem Physiol, 2018, 147:83-89.
[24]兰杰,司乃国,李志念,等. SYP-14288与百菌清、代森锰锌生物活性的比较[J].农药, 2012, 51(12):927-928.LAN J, SI N G, LI Z N, et al. Comparative of the biological activities of SYP-14288, chlorothalonil and mancozeb[J]. Agrochemicals,2012, 51(12):927-928.
[25]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 SYP-14288 against cucumber downy mildew[M]//Chemical control of plant diseases in China:Vol. 9, 2014:51-57.
[26]兰杰,李志念,司乃国,等. SYP-14288室内生物活性对黄瓜霜霉田间药效研究[C]//中国化工学会农药专业委员会第十五届年会论文集, 2012:391-397.LAN J, LI Z N, SI N G, et al. The lab and field efficacy of SYP-14288 against cucumber downy mildew[C]//Proceedings of the 15th annual meeting of the pesticide professional committee of the Chinese chemical society, 2012:391-397.
[27]侯燕华,彭钦,方媛,等.辣椒疫霉对双苯菌胺抗性风险评估[C].中国植物病理学会2015年学术年会论文集.北京:中国农业科学技术出版社, 2015:122.HOU Y H, PENG Q, FANG Y, et al. The resistance risk of Phytophthora capsici to SYP-14288[C]//Proceedings of the annual meeting of Chinese Society of Plant Pathology in 2011. Beijing:China Agricultural Science and Technology Press, 2015:122.
[28]王佳韵,张一宾.全球三唑类杀菌剂的市场概况及发展趋势[J].农药, 2017, 56(2):86-87.WANG J Y, ZHANG Y B. Market and development tendency of global triazole fungicides[J]. Agrochemicals, 2017, 56(2):86-87.
[2]DIXON L J, SCHLUB R L, PERNEZNY K, et al. Host specialization and phylogenetic diversity of Corynespora cassiicola[J].Phytopathology, 2009, 99(9):1015-1027.
[3]李长松,张眉,李林,等.山东省黄瓜棒孢叶斑病(褐斑病)病原菌鉴定和防治[J].中国蔬菜, 2009, 1(18):29-33.LI C S, ZHANG M, LI L, et al. Identification of cucumber target leaf spot(brown spot)pathogen and its control[J]. China Vegetables,2009, 1(18):29-33.
[4]禾丽菲,李晓旭,朱佳美,等.不同杀菌剂对黄瓜靶斑病菌的毒力作用特性比较[J].农药学学报, 2018, 20(1):25-32.HE L F, LI X X, ZHU J M, et al. Comparison of toxicity properties of different types of fungicides against Corynespora cassiicola on cucumber[J]. Chin J Pestic Sci, 2018, 20(1):25-32.
[5]KWON M K, KANG B R, CHO B H, et al. Occurrence of target leaf spot disease caused by Corynespora cassicola on cucumber in Korea[J]. Plant Pathol, 2003, 52(3):424.
[6]蓝国兵,谭耀华,何自福,等.黄瓜褐斑病(Corynespora cassiicola)在广东首次报道[J].植物保护, 2012, 38(5):197-200.LAN G B, TAN Y H, HE Z F, et al. First report of Cucumis sativus brown spot disease infected by Corynespora cassiicola in Guangdong Province[J]. Plant Prot, 2012, 38(5):197-200.
[7]杜玉宁,查仙芳,杨金凤,等.宁夏温室黄瓜新病害靶斑病的药剂筛选[J].江苏农业科学, 2013, 41(1):132-133.DU Y N, ZHA X F, YANG J F, et al. Screening of Corynespora cassiicolaon cucumber in greenhouse of Ningxia[J]. Jiangsu Agricultural Sciences, 2013, 41(1):132-133.
[8]杨双娟,顾兴芳,张圣平,等.黄瓜棒孢叶斑病(Corynespora cassiicola)的研究概况[J].中国蔬菜, 2012, 1(4):1-9.YANG S J, GU X F, ZHANG S P, et al. Research progress on cucumber target leaf spot(Corynespora cassiicola)[J]. China Vegetables, 2012, 1(4):1-9.
[9]张乃楼,李亚美,康文强,等.辽宁省黄瓜靶斑病菌对苯醚甲环唑和戊唑醇的敏感性[J].农药学学报, 2014, 16(4):452-456.ZHANG N L, LI Y M, KANG W Q, et al. Sensitivity of Corynespora cassiicolato difenoconazole and tebuconazole in Liaoning Province[J]. Chin J Pestic Sci, 2014, 16(4):452-456.
[10]张中义,李继新,关国经,等.烤烟棒孢霉叶斑病病原菌鉴定[J].中国烟草学报, 2008, 14(6):44-47.ZHANG Z Y, LI J X, GUAN G J, et al. Identification of fungal pathogen of corynespora leaf spot in flue-cured tobacco[J]. Acta Tabacaria Sinica, 2008, 14(6):44-47.
[11]XUE L H, LIU Y, LI C J. First report of leaf spot caused by Corynespora cassiicola on Ixeris polycephala in China[J]. Plant Dis,2017, 101(8):1552.
[12]YEH Y H, KIRSCHNER R. First report of Corynespora leaf spot on Vitex rotundifolia caused by Corynespora cassiicola in Taiwan[J].Plant Dis, 2017, 101(8):1550.
[13]BAIYEE B, PORNSURIYA C, ITO S I, et al. Trichoderma spirale T76-1 displays biocontrol activity against leaf spot on lettuce(Lactuca sativa L.)caused by Corynespora cassiicola or Curvularia aeria[J]. Biol Control, 2019, 129:195-200.
[14]BUTLER S, YOUNG-KELLY H, RAPER T, et al. First report of target spot caused by Corynespora cassiicolaon cotton in Tennessee[J]. Plant Dis, 2016, 100(2):535.
[15]HUANG X, ZHENG X, XU J, et al. First report of brown leaf spot on honeysuckle caused by Corynespora cassiicola in China[J]. Plant Dis,2016, 100(11):2326.
[16]XU J, QI X, ZHENG X. First report of Corynespora leaf spot on sweet potato caused by Corynespora cassiicolain China[J].Australasian Plant Disease Notes, 2016, 100(10):2163.
[17]ONOFRE R B, MERTELY J C, AGUIAR F M, et al. First report of target spot caused by Corynespora cassiicola on blueberry in North America[J]. Plant Dis, 2016, 100(2):528-529.
[18]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.
[19]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.
[20]WHITE T J, BRUNS T, LEE S, et al. 38 amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[J]. Pcr Protoc, 1990:315-322.
[21]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, 1994:76-81.
[22]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.
[23]WANG Z W, NI X X, PENG Q, et al. The novel fungicide SYP-14288 acts as an uncoupler against Phytophthora capsici[J]. Pestic Biochem Physiol, 2018, 147:83-89.
[24]兰杰,司乃国,李志念,等. SYP-14288与百菌清、代森锰锌生物活性的比较[J].农药, 2012, 51(12):927-928.LAN J, SI N G, LI Z N, et al. Comparative of the biological activities of SYP-14288, chlorothalonil and mancozeb[J]. Agrochemicals,2012, 51(12):927-928.
[25]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 SYP-14288 against cucumber downy mildew[M]//Chemical control of plant diseases in China:Vol. 9, 2014:51-57.
[26]兰杰,李志念,司乃国,等. SYP-14288室内生物活性对黄瓜霜霉田间药效研究[C]//中国化工学会农药专业委员会第十五届年会论文集, 2012:391-397.LAN J, LI Z N, SI N G, et al. The lab and field efficacy of SYP-14288 against cucumber downy mildew[C]//Proceedings of the 15th annual meeting of the pesticide professional committee of the Chinese chemical society, 2012:391-397.
[27]侯燕华,彭钦,方媛,等.辣椒疫霉对双苯菌胺抗性风险评估[C].中国植物病理学会2015年学术年会论文集.北京:中国农业科学技术出版社, 2015:122.HOU Y H, PENG Q, FANG Y, et al. The resistance risk of Phytophthora capsici to SYP-14288[C]//Proceedings of the annual meeting of Chinese Society of Plant Pathology in 2011. Beijing:China Agricultural Science and Technology Press, 2015:122.
[28]王佳韵,张一宾.全球三唑类杀菌剂的市场概况及发展趋势[J].农药, 2017, 56(2):86-87.WANG J Y, ZHANG Y B. Market and development tendency of global triazole fungicides[J]. Agrochemicals, 2017, 56(2):86-87.