茶角胸叶甲高毒力球孢白僵菌菌株的筛选
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摘要
茶角胸叶甲(Basilepta melanopus Lefevre)是一种茶树上常见的食叶性害虫,其发生为害严重影响茶叶的产量和品质。近年来,该虫在局部茶园,尤其是有机茶园中常爆发成灾。如何有效防治该害虫,已成为亟待解决的问题。本研究选择原寄主为茶园鞘翅目害虫的9个球孢白僵菌(Beauveria bassiana)菌株,依据菌落形态、菌株生长速率、产孢量、孢子萌发率、耐热力和抗紫外能力等生物学性状初步筛选出Bb338、Bb346和Bb2-1等3个生物学性状优良的菌株;在此基础上,利用浸虫法研究了3个菌株对茶角胸叶甲成虫的杀虫毒力。生测结果表明,用每毫升1×10~7个孢子的孢悬液处理后,球孢白僵菌Bb2-1对茶角胸叶甲成虫的累计校正死亡率达100%,僵虫率达86.11%;LT_(50)仅为3.32 d。综合上述各生物学特性和杀虫毒力表明,筛选到的Bb2-1菌株具有生长速率快、产孢量大、耐热力和抗紫外能力强,以及对茶角胸叶甲杀虫毒力最强和杀虫速率最快等优点,在今后茶角胸叶甲的生物防治中将发挥重要的作用。
The beetle(Basilepta melanopus) is one of the most serious leaf-feeding insects in tea garden, which seriously affects both tea yield and quality. In recent years, the beetle often caused serious damage in some tea gardens, especially organic tea gardens. How to effectively control this pest has become a serious problem. In order to screen high virulence strains of Beauveria bassiana to B. melanopus, 9 strains of B. bassiana isolated from the coleoptera pests in tea gardens were selected as candidates. In the preliminary screening, the biological characteristics of these 9 strains, including colony morphology, growth rate, sporulation, spore germination rate, thermotolerance and UV radiation resistance were detected. According to the above biological characteristics, three strains Bb338, Bb346 and Bb2-1 were selected for further bioassay against the adult of B. melanopus. Bioassay results showed that the strain Bb2-1 was the most virulent strain to the adults of B. melanopus, which had the highest corrected mortality rates of 100%, highest cadaver rates of 86.11% and shortest LT_(50) of 3.32 d at a concentration of 1.0×10~7 conidia per milliliter. The strain Bb2-1 has the best biological characteristics and the highest virulent against the adults of B. melanopus, which would play a very important role in biocontrol of B. melanopus.
The beetle(Basilepta melanopus) is one of the most serious leaf-feeding insects in tea garden, which seriously affects both tea yield and quality. In recent years, the beetle often caused serious damage in some tea gardens, especially organic tea gardens. How to effectively control this pest has become a serious problem. In order to screen high virulence strains of Beauveria bassiana to B. melanopus, 9 strains of B. bassiana isolated from the coleoptera pests in tea gardens were selected as candidates. In the preliminary screening, the biological characteristics of these 9 strains, including colony morphology, growth rate, sporulation, spore germination rate, thermotolerance and UV radiation resistance were detected. According to the above biological characteristics, three strains Bb338, Bb346 and Bb2-1 were selected for further bioassay against the adult of B. melanopus. Bioassay results showed that the strain Bb2-1 was the most virulent strain to the adults of B. melanopus, which had the highest corrected mortality rates of 100%, highest cadaver rates of 86.11% and shortest LT_(50) of 3.32 d at a concentration of 1.0×10~7 conidia per milliliter. The strain Bb2-1 has the best biological characteristics and the highest virulent against the adults of B. melanopus, which would play a very important role in biocontrol of B. melanopus.
引文
[1]张汉鹄,谭济才.中国茶树害虫及其无公害治理[M].合肥:安徽科学技术出版社,2004:241-242.
[2]羊柏娥,王沅江.茶角胸叶甲研究进展[J].茶叶通讯,2008,35(3):24-26.
[3]曾明森,吴光远.福建茶树病虫害与天敌图谱[M].北京:中国农业科学技术出版社,2014:83-85.
[4]何学友,蔡守平,吴智才.油茶的一种新害虫——黑足角胸叶甲[J].中国森林病虫,2011(3):16-17.
[5]汪荣灶.黑足角胸叶甲发生与危害的初步考查[J].福建茶叶,1989(2):40-41.
[6]汪荣灶,胡勇.江西黑足角胸叶甲成虫的盛发期与防治[J].江西植保,2004,27(4):175.
[7]谭济才,刘贵芳,王德兴,等.茶角胸叶甲的发生与防治[J].中国茶叶,1987,9(1):15-16.
[8]林阿祥.茶角胸叶甲的发生与防治初报[J].茶叶科学技术,1991(4):43-44.
[9]谭济才,刘贵芳,王德兴,等.茶角胸叶甲生物学特性及防治研究[J].湖南农学院学报,1986(4):51-60.
[10]李先文,谭济才,陈建芝,等.5%锐劲特悬浮剂防治茶角胸叶甲试验[J].现代农业科技,2007(1):52.
[11]李先文,谭济才,柏晓勇,等.几种药剂对茶角胸叶甲的室内杀虫活性测定及田间药效试验[J].现代农药,2008,7(3):44-47.
[12]蒲蛰龙,李增智.昆虫真菌学[M].合肥:安徽科学技术出版社,1996:206-360.
[13]Faria MR de,Wraight SP.Mycoinsecticides and Mycoacaricides:A comprehensive list with worldwide coverage and international classification of formulation types[J].Biology Control,2007,43:237-256.
[14]林乃铨.害虫生物防治[M].北京:科学出版社,2010:239-265.
[15]李增智,黄勃,陈名君,等.分子时代的白僵菌研究[J].菌物学报,2011,30(6):823-835.
[16]何学友,蔡守平,童应华,等.球孢白僵菌和金龟子绿僵菌不同菌株对黑足角胸叶甲成虫的致病力评价[J].昆虫学报,2011,54(11):1281-1287.
[17]曾明森,刘丰静,王定锋,等.茶角胸叶甲综合治理试验示范[J].福建农业学报,2012,27(8):847-852.
[18]Poprawski TJ,Marchal M,Robert PH.Comparative susceptibility of Otiorhynchus sulcatus and Sitona lineatus(Coleoptera:Curculionidae)early stage to five entomopathogenic hyphomycetes[J].Environmental Entomology,1985,14:247-253.
[19]Soares GG,Marchal M,Ferron P.Susceptibility of Otiorhynchus sulcatus(Coleoptera:Curculionidae)larvae to Metarhizium anisopliae and Metarhizium flavoviridae(Deuteromycotina:Hyphomycetes)at two different temperatures[J].Environmental Entomology,1983,12:1886-1890.
[20]王定锋,黎健龙,李慧玲,等.茶丽纹象甲白僵菌广东分离株的鉴定及生物学特性研究[J].茶叶科学,2015,35(5):449-457.
[21]雷妍圆,吕利华,何余容,等.球孢白僵菌生物学特性与其对小菜蛾致病力相关性分析[J].中国生物防治,2010,26(2):143-148.
[22]俞佳,冯明光.基于分生孢子热胁迫反应的球孢白僵菌耐热菌株筛选[J].菌物学报,2006,25(2):278-283.
[23]许齐爱,李佳颖,任顺祥.红火蚁高致病力黄绿绿僵菌菌株的筛选[J].中国生物防治学报,2013,29(3):384-388.
[24]王定锋,黎健龙,王庆森,等.柑橘灰象甲一株高毒力白僵菌菌株的筛选鉴定及培养特性[J].中国生物防治学报,2014,30(6):750-758.
[25]侯明明,刘义豪,王滨.昆虫病原真菌对梨网蝽防治潜力的室内评测[J].中国生物防治学报,2015,31(6):853-859.
[26]刘玉军,张龙娃,何亚琼,等.栎旋木柄天牛高毒力球孢白僵菌菌株的筛选[J].昆虫学报,2008,51(2):143-149.
[27]张龙娃,康克,刘玉军,等.美国白蛾高毒力球孢白僵菌菌株筛选[J].昆虫学报,2016,59(1):111-118.
[28]Zhang LW,Liu YJ,Yao J,et al.Evaluation of Beauveria bassiana(Hyphomycetes)isolates as potential agents for control of Dendroctonus valens[J].Insect Science,2011,18:209-216.
[29]Luz C,Fargues J.Temperature and moisture requirements for conidial germination of an isolate of Beauveria bassiana,pathogenic to Rhodnius prolixus[J].Mycopathology,1997,138:117-125.
[30]Devi KU,Sridevi V,Challa MM,et al.Effect of high temperature and water stress on in vitro germination and growth in isolates of the entomopathogenic fungus Beauveria bassiana(Bals.)Vuillemin[J].J Invertebr Pathol,2005,88:181-189.
[31]Fargues J,Goettle MS,Smits N,et al.Effect of temperature on vegetative growth of Beauveria bassiana isolates from different origins[J].Mycologia,1997,89:383-392.
[32]孙鲁娟,吴孔明,郭予元,不同温、湿度下白僵菌对棉铃虫幼虫的致病力[J].昆虫学报,2001,44(4):501-506.
[33]何余容,邝灼彬,冯夏,等.不同温湿度下球孢白僵菌对小猿叶甲的致病力[J].昆虫学报,2005,48(5):679-686
[34]Hong TD,Gunn J,Ellis RH,et al.The effect of storage environment on the longevity of conidia of Beauveria bassiana[J].Mycological Research,2001,105:597-602.
[35]李鸿文,冯明光.球孢白僵菌不同菌株分生孢子的耐热能力[J].浙江大学学报:农业与生命科学版,2008,34(2):158-162.
[36]Ignoffo CM,Garcia C.Influence of conidial color on inactivation of several entomogenous fungi(Hyphomycetes)by simulated sunlight[J].Environ Entomol,1992,21:913–917.
[37]Inglis GD,Goettel MS,Johnson DL.Influence of ultraviolet light protectants on persistence of the entomopathogenic fungus,Beauveria bassiana[J].Biological Control,1995,5(4):581-590.
[38]Fargus J,Goettel MS,Smits N,et al.Variability in susceptibility to simulated sunlight of conidia among isolates of entomopathogenic Hyphomycetes[J].Mycopathologia,1996,135:171-181.
[2]羊柏娥,王沅江.茶角胸叶甲研究进展[J].茶叶通讯,2008,35(3):24-26.
[3]曾明森,吴光远.福建茶树病虫害与天敌图谱[M].北京:中国农业科学技术出版社,2014:83-85.
[4]何学友,蔡守平,吴智才.油茶的一种新害虫——黑足角胸叶甲[J].中国森林病虫,2011(3):16-17.
[5]汪荣灶.黑足角胸叶甲发生与危害的初步考查[J].福建茶叶,1989(2):40-41.
[6]汪荣灶,胡勇.江西黑足角胸叶甲成虫的盛发期与防治[J].江西植保,2004,27(4):175.
[7]谭济才,刘贵芳,王德兴,等.茶角胸叶甲的发生与防治[J].中国茶叶,1987,9(1):15-16.
[8]林阿祥.茶角胸叶甲的发生与防治初报[J].茶叶科学技术,1991(4):43-44.
[9]谭济才,刘贵芳,王德兴,等.茶角胸叶甲生物学特性及防治研究[J].湖南农学院学报,1986(4):51-60.
[10]李先文,谭济才,陈建芝,等.5%锐劲特悬浮剂防治茶角胸叶甲试验[J].现代农业科技,2007(1):52.
[11]李先文,谭济才,柏晓勇,等.几种药剂对茶角胸叶甲的室内杀虫活性测定及田间药效试验[J].现代农药,2008,7(3):44-47.
[12]蒲蛰龙,李增智.昆虫真菌学[M].合肥:安徽科学技术出版社,1996:206-360.
[13]Faria MR de,Wraight SP.Mycoinsecticides and Mycoacaricides:A comprehensive list with worldwide coverage and international classification of formulation types[J].Biology Control,2007,43:237-256.
[14]林乃铨.害虫生物防治[M].北京:科学出版社,2010:239-265.
[15]李增智,黄勃,陈名君,等.分子时代的白僵菌研究[J].菌物学报,2011,30(6):823-835.
[16]何学友,蔡守平,童应华,等.球孢白僵菌和金龟子绿僵菌不同菌株对黑足角胸叶甲成虫的致病力评价[J].昆虫学报,2011,54(11):1281-1287.
[17]曾明森,刘丰静,王定锋,等.茶角胸叶甲综合治理试验示范[J].福建农业学报,2012,27(8):847-852.
[18]Poprawski TJ,Marchal M,Robert PH.Comparative susceptibility of Otiorhynchus sulcatus and Sitona lineatus(Coleoptera:Curculionidae)early stage to five entomopathogenic hyphomycetes[J].Environmental Entomology,1985,14:247-253.
[19]Soares GG,Marchal M,Ferron P.Susceptibility of Otiorhynchus sulcatus(Coleoptera:Curculionidae)larvae to Metarhizium anisopliae and Metarhizium flavoviridae(Deuteromycotina:Hyphomycetes)at two different temperatures[J].Environmental Entomology,1983,12:1886-1890.
[20]王定锋,黎健龙,李慧玲,等.茶丽纹象甲白僵菌广东分离株的鉴定及生物学特性研究[J].茶叶科学,2015,35(5):449-457.
[21]雷妍圆,吕利华,何余容,等.球孢白僵菌生物学特性与其对小菜蛾致病力相关性分析[J].中国生物防治,2010,26(2):143-148.
[22]俞佳,冯明光.基于分生孢子热胁迫反应的球孢白僵菌耐热菌株筛选[J].菌物学报,2006,25(2):278-283.
[23]许齐爱,李佳颖,任顺祥.红火蚁高致病力黄绿绿僵菌菌株的筛选[J].中国生物防治学报,2013,29(3):384-388.
[24]王定锋,黎健龙,王庆森,等.柑橘灰象甲一株高毒力白僵菌菌株的筛选鉴定及培养特性[J].中国生物防治学报,2014,30(6):750-758.
[25]侯明明,刘义豪,王滨.昆虫病原真菌对梨网蝽防治潜力的室内评测[J].中国生物防治学报,2015,31(6):853-859.
[26]刘玉军,张龙娃,何亚琼,等.栎旋木柄天牛高毒力球孢白僵菌菌株的筛选[J].昆虫学报,2008,51(2):143-149.
[27]张龙娃,康克,刘玉军,等.美国白蛾高毒力球孢白僵菌菌株筛选[J].昆虫学报,2016,59(1):111-118.
[28]Zhang LW,Liu YJ,Yao J,et al.Evaluation of Beauveria bassiana(Hyphomycetes)isolates as potential agents for control of Dendroctonus valens[J].Insect Science,2011,18:209-216.
[29]Luz C,Fargues J.Temperature and moisture requirements for conidial germination of an isolate of Beauveria bassiana,pathogenic to Rhodnius prolixus[J].Mycopathology,1997,138:117-125.
[30]Devi KU,Sridevi V,Challa MM,et al.Effect of high temperature and water stress on in vitro germination and growth in isolates of the entomopathogenic fungus Beauveria bassiana(Bals.)Vuillemin[J].J Invertebr Pathol,2005,88:181-189.
[31]Fargues J,Goettle MS,Smits N,et al.Effect of temperature on vegetative growth of Beauveria bassiana isolates from different origins[J].Mycologia,1997,89:383-392.
[32]孙鲁娟,吴孔明,郭予元,不同温、湿度下白僵菌对棉铃虫幼虫的致病力[J].昆虫学报,2001,44(4):501-506.
[33]何余容,邝灼彬,冯夏,等.不同温湿度下球孢白僵菌对小猿叶甲的致病力[J].昆虫学报,2005,48(5):679-686
[34]Hong TD,Gunn J,Ellis RH,et al.The effect of storage environment on the longevity of conidia of Beauveria bassiana[J].Mycological Research,2001,105:597-602.
[35]李鸿文,冯明光.球孢白僵菌不同菌株分生孢子的耐热能力[J].浙江大学学报:农业与生命科学版,2008,34(2):158-162.
[36]Ignoffo CM,Garcia C.Influence of conidial color on inactivation of several entomogenous fungi(Hyphomycetes)by simulated sunlight[J].Environ Entomol,1992,21:913–917.
[37]Inglis GD,Goettel MS,Johnson DL.Influence of ultraviolet light protectants on persistence of the entomopathogenic fungus,Beauveria bassiana[J].Biological Control,1995,5(4):581-590.
[38]Fargus J,Goettel MS,Smits N,et al.Variability in susceptibility to simulated sunlight of conidia among isolates of entomopathogenic Hyphomycetes[J].Mycopathologia,1996,135:171-181.