山东蓬莱葡萄灰霉菌对7种杀菌剂的抗药性检测
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摘要
为了明确葡萄灰霉菌对啶酰菌胺、多菌灵、咯菌腈、异菌脲、腐霉利、嘧霉胺的抗药性和对抑霉唑的敏感性,本试验采用菌丝生长速率法和孢子萌发法检测了采自山东蓬莱地区的69株葡萄灰霉菌对上述前6种杀菌剂的抗药性、对抑霉唑的敏感性及抑霉唑与其他6种杀菌剂的交互抗性关系。结果表明,抑霉唑对这69株葡萄灰霉菌的EC_(50)分布在0.403～28.76μg/mL之间,平均值为(9.34±10.34)μg/mL;葡萄灰霉菌菌株中抗啶酰菌胺(BosR)、多菌灵(CarR)、咯菌腈(FluR)、异菌脲(IprR)、嘧霉胺(PyrR)、腐霉利(ProR)的比例分别为100%、100%、9.47%、97.18%、100%、89.20%,测试菌株的抗药性均为多抗类型,没有单抗菌株,其中对3种杀菌剂(啶酰菌胺、多菌灵、嘧霉胺)、对4种杀菌剂(啶酰菌胺、多菌灵、异菌脲、嘧霉胺)、对5种杀菌剂(啶酰菌胺、多菌灵、异菌脲、嘧霉胺、腐霉利或啶酰菌胺、多菌灵、咯菌腈、异菌脲、嘧霉胺)和对6种杀菌剂(啶酰菌胺、多菌灵、咯菌腈、异菌脲、嘧霉胺、腐霉利)的抗性频率分别为2.33%、9.30%、79.07%、2.33%、6.97%,表明啶酰菌胺、多菌灵、嘧霉胺对测试葡萄菌株完全丧失防效,建议在该葡萄产区停止使用这些药剂,测试菌株对腐霉利、异菌脲的抗性频率高,建议采取限制使用、禁止单独使用等措施,测试菌株对咯菌腈的抗性频率较低,可以继续使用但需按照科学使用规则进行。抑霉唑与其他6种杀菌剂间不存在交互抗性关系,说明其可以和其他药剂同时使用但建议减少使用。
In order to investigate the resistance of Botrytis cinerea to carbendazim, procymidone, iprodione, fludioxonil, pyrimethanil and boscalid, the sensitivity to imazalil, and the cross-resistance between imazalil and other six fungicides, 69 isolates of B.cinerea from grape plants were collected from the vine planting areas in Penglai of Shandong Province and bioassay was implemented by the method of mycelial growth and conidial germination inhibition. The results indicated that the mean EC_(50) values of 69 isolates to imazalil were(9.34±10.34)μg/mL, ranging from 0.403 to 28.76 μg/mL. The resistance frequencies of tested isolates to CarR, BosR and PyrR were 100%,those of to ProR and IprR were above 85%, and that of to FluR was 9.47%. The multidrug resistance types were PyrR_BosR_CarR, IprR_PyrR_BosR_CarR, IprR_PyrR_FluR_BosR_CarR, IprR_PyrR_ProR BosR_CarR and IprR_PyrR_FluR_BosR_CarR_ProR,accounting for 2.33%, 9.30%, 2.33%,79.07% and 6.97%, respectively. The results showed that boscalid, carbendazim, pyrimethanil could not effectively control gray mold, so we should stop applying these fungicides in vineyard of Penglai. As to procymidone and iprodione, strategies must be implemented which prevent any possibilities for solo application and use as little as possible. Fludioxonil should be selected and used for controlling Botrytis rot according to general use recommendations. Imazalil does not have cross-resistance with other six fungicides, and it has lower activity to Botrytis cinerea, so we should not select imazalil for controlling Botrytis rot in vineyard if there are many effective fungicides for choice.
In order to investigate the resistance of Botrytis cinerea to carbendazim, procymidone, iprodione, fludioxonil, pyrimethanil and boscalid, the sensitivity to imazalil, and the cross-resistance between imazalil and other six fungicides, 69 isolates of B.cinerea from grape plants were collected from the vine planting areas in Penglai of Shandong Province and bioassay was implemented by the method of mycelial growth and conidial germination inhibition. The results indicated that the mean EC_(50) values of 69 isolates to imazalil were(9.34±10.34)μg/mL, ranging from 0.403 to 28.76 μg/mL. The resistance frequencies of tested isolates to CarR, BosR and PyrR were 100%,those of to ProR and IprR were above 85%, and that of to FluR was 9.47%. The multidrug resistance types were PyrR_BosR_CarR, IprR_PyrR_BosR_CarR, IprR_PyrR_FluR_BosR_CarR, IprR_PyrR_ProR BosR_CarR and IprR_PyrR_FluR_BosR_CarR_ProR,accounting for 2.33%, 9.30%, 2.33%,79.07% and 6.97%, respectively. The results showed that boscalid, carbendazim, pyrimethanil could not effectively control gray mold, so we should stop applying these fungicides in vineyard of Penglai. As to procymidone and iprodione, strategies must be implemented which prevent any possibilities for solo application and use as little as possible. Fludioxonil should be selected and used for controlling Botrytis rot according to general use recommendations. Imazalil does not have cross-resistance with other six fungicides, and it has lower activity to Botrytis cinerea, so we should not select imazalil for controlling Botrytis rot in vineyard if there are many effective fungicides for choice.
引文
[1] DEAN R, VAN KAN J A, PRETORIUS Z A, et al. The top 10 fungal pathogens in molecular plant pathology [J]. Molecular Plant Pathology, 2012, 13(4): 804.
[2] 张玮,乔广行,黄金宝,等. 中国葡萄灰霉病菌对嘧霉胺的抗药性检测[J]. 中国农业科学,2013, 46(6): 1208-1212.
[3] RODRíGUEZ A, ACOSTA A, RODRíGUEZ C . Fungicide resistance of Botrytis cinerea in tomato greenhouses in the Canary Islands and effectiveness of non-chemical treatments against gray mold [J]. World Journal of Microbiology & Biotechnology, 2014, 30(9):2397-2406.
[4] 尹大芳. 浙江省草莓灰霉病菌抗药性检测及抗性机制的研究[D]. 杭州:浙江大学, 2015.
[5] SABRINA R , WEBER R W S , DANIEL R , et al. Spread of Botrytis cinerea strains with multiple fungicide resistance in German horticulture[J]. Frontiers in Microbiology, 2017, 7:2075.
[6] MADE W. Synergism and antagonism in fungicide mixtures containing sterol demethylation inhibitors[J]. Phytopathology, 1996, 86: 1280-1283.
[7] GREENBERG R, C RESNICK C. A gas liquid chromatographic method for determining imazalil residues in citrus fruit[J]. Pest Management Science, 2010, 8(1): 59-64.
[8] BUS V G. ED50 levels of Penicillium digitatum and P. italicum with reduced sensitivity to thiabendazole, benomyl and imazalil [J]. Postharvest Biology & Technology, 1992, 1(4): 305-315.
[9] BUS V G, BONGERS A J, RISSE L A. Occurrence of Penicillium digitatum and P. italicum resistant to benomyl, thiabendazole, and imazalil on citrus fruit from different geographic origins [J]. Plant Disease, 1991, 75: 1098-1100.
[10] ZHU Jinwen, XIE Qingyun, LI Hongye. Occurrence of imazalil resistant biotype of Penicillium digitatum in China and the resistant molecular mechanism [J]. Journal of Zhejiang University Science A, 2006, 7(2): 362-365.
[11] WIGLER P W, PATTERSON F K. Inhibition of the multidrug resistance efflux pump [J]. Biochimica et Biophysica Acta, 1993, 1154(2): 173-181.
[12] 黄邦彦, 吴立. 柑橘贮藏防腐剂抑霉唑的引进和应用试验[J]. 中国南方果树, 1982 (4): 19-20.
[13] YIN Dafang, CHEN Xiang, MA Zhonghua, et al. Multiple resistance to QOIs and other classes of fungicides in Botrytis cinerea populations from strawberry in Zhejiang Province, China [J]. European Journal of Plant Pathology, 2015, 141(1): 169-177.
[14] BARDAS G A, MYRESIOTIS C K, KARAOGLANIDIS G S. Stability and fitness of anilinopyrimidine-resistant strains of Botrytis cinerea [J]. Phytopathology,2008,98(4):443-450.
[15] MYRESIOTIS C, KARAOGLANIDIS G, TZAVELLA-KLONARI K. Resistance of Botrytis cinerea isolates from vegetable crops to anilinopyrimidine, phenylpyrrole, hydroxyanilide, benzimidazole, and dicarboximide fungicide [J]. Plant Disease, 2007, 91(4): 407-413.
[16] 乔广行,林秀敏,等. 8种杀菌剂对番茄灰霉病菌多重抗药性菌株生物活性测定[J]. 农药, 2013, 52(1): 57-59.
[17] SUN Haiyan, WANG Hancheng, CHEN Yu, et al. Multiple resistance of Botrytis cinerea from vegetable crops to carbendazim, diethofencarb, procymidone and pyrimethanil in China[J]. Plant Disease, 2010, 94(5): 551-556.
[18] FERNANDEZ-ORTUNO D, CHEN F P, SCHNABEL G. Resistance to cyprodinil and lack of fludioxonil resistance in Botrytis cinerea isolates from strawberry in North and South Carolina [J]. Plant Disease, 2013, 97(1): 81-85.
[2] 张玮,乔广行,黄金宝,等. 中国葡萄灰霉病菌对嘧霉胺的抗药性检测[J]. 中国农业科学,2013, 46(6): 1208-1212.
[3] RODRíGUEZ A, ACOSTA A, RODRíGUEZ C . Fungicide resistance of Botrytis cinerea in tomato greenhouses in the Canary Islands and effectiveness of non-chemical treatments against gray mold [J]. World Journal of Microbiology & Biotechnology, 2014, 30(9):2397-2406.
[4] 尹大芳. 浙江省草莓灰霉病菌抗药性检测及抗性机制的研究[D]. 杭州:浙江大学, 2015.
[5] SABRINA R , WEBER R W S , DANIEL R , et al. Spread of Botrytis cinerea strains with multiple fungicide resistance in German horticulture[J]. Frontiers in Microbiology, 2017, 7:2075.
[6] MADE W. Synergism and antagonism in fungicide mixtures containing sterol demethylation inhibitors[J]. Phytopathology, 1996, 86: 1280-1283.
[7] GREENBERG R, C RESNICK C. A gas liquid chromatographic method for determining imazalil residues in citrus fruit[J]. Pest Management Science, 2010, 8(1): 59-64.
[8] BUS V G. ED50 levels of Penicillium digitatum and P. italicum with reduced sensitivity to thiabendazole, benomyl and imazalil [J]. Postharvest Biology & Technology, 1992, 1(4): 305-315.
[9] BUS V G, BONGERS A J, RISSE L A. Occurrence of Penicillium digitatum and P. italicum resistant to benomyl, thiabendazole, and imazalil on citrus fruit from different geographic origins [J]. Plant Disease, 1991, 75: 1098-1100.
[10] ZHU Jinwen, XIE Qingyun, LI Hongye. Occurrence of imazalil resistant biotype of Penicillium digitatum in China and the resistant molecular mechanism [J]. Journal of Zhejiang University Science A, 2006, 7(2): 362-365.
[11] WIGLER P W, PATTERSON F K. Inhibition of the multidrug resistance efflux pump [J]. Biochimica et Biophysica Acta, 1993, 1154(2): 173-181.
[12] 黄邦彦, 吴立. 柑橘贮藏防腐剂抑霉唑的引进和应用试验[J]. 中国南方果树, 1982 (4): 19-20.
[13] YIN Dafang, CHEN Xiang, MA Zhonghua, et al. Multiple resistance to QOIs and other classes of fungicides in Botrytis cinerea populations from strawberry in Zhejiang Province, China [J]. European Journal of Plant Pathology, 2015, 141(1): 169-177.
[14] BARDAS G A, MYRESIOTIS C K, KARAOGLANIDIS G S. Stability and fitness of anilinopyrimidine-resistant strains of Botrytis cinerea [J]. Phytopathology,2008,98(4):443-450.
[15] MYRESIOTIS C, KARAOGLANIDIS G, TZAVELLA-KLONARI K. Resistance of Botrytis cinerea isolates from vegetable crops to anilinopyrimidine, phenylpyrrole, hydroxyanilide, benzimidazole, and dicarboximide fungicide [J]. Plant Disease, 2007, 91(4): 407-413.
[16] 乔广行,林秀敏,等. 8种杀菌剂对番茄灰霉病菌多重抗药性菌株生物活性测定[J]. 农药, 2013, 52(1): 57-59.
[17] SUN Haiyan, WANG Hancheng, CHEN Yu, et al. Multiple resistance of Botrytis cinerea from vegetable crops to carbendazim, diethofencarb, procymidone and pyrimethanil in China[J]. Plant Disease, 2010, 94(5): 551-556.
[18] FERNANDEZ-ORTUNO D, CHEN F P, SCHNABEL G. Resistance to cyprodinil and lack of fludioxonil resistance in Botrytis cinerea isolates from strawberry in North and South Carolina [J]. Plant Disease, 2013, 97(1): 81-85.