一株茶大灰象甲寄生真菌的分子鉴定及其毒力测定
|
摘要
为明确分离自茶大灰象甲(Sympiezomias citri)自然羅病死亡成虫僵虫的一株寄生真菌Bbr4941菌株的具体分类归属及其生防潜力,对该菌株的rDNA-ITS序列和Bloc序列进行了PCR扩增、测序及系统进化树构建。结果明确了它为布氏白僵菌(Beauveria brongniartii)。在此基础上,利用浸虫法研究该菌株对茶大灰象甲成虫的室内毒力。结果表明,高浓度(每毫升5.0×10~7孢子)孢悬液处理时,第7天累计校正死亡率为100%,LT_(50)为3.916 d。以上研究表明Bbr4941菌株对茶大灰象甲成虫具有很强的杀虫毒力,有望在今后茶大灰象甲的生物防治中发挥重要作用。
The taxonomic status and biocontrol potential of an entomopathogenic fungus that isolated from the cadaver of Sympiezomias citri were investigated. Amplification, sequencing, and system evolution tree construction of the nuclear ribosomal internal transcribed spacer(rDNA-ITS) and intergenic Bloc region fragments(Bloc) of the fungus Bbr4941, and the bioassay against the adult of S. citri were carried out. The results showed that the entomopathogenic fungus was identified as Beauveria brongniartii. When the adults of S. citri were dealt with Bbr4941 suspension(5.0×10~7 spore·mL~(-1)), the corrected accumulative mortality was 100% and LT_(50) value was 3.916 d after 7 days. Our results showed that Bbr4941 had great potential for the biocontrol of S. citri.
The taxonomic status and biocontrol potential of an entomopathogenic fungus that isolated from the cadaver of Sympiezomias citri were investigated. Amplification, sequencing, and system evolution tree construction of the nuclear ribosomal internal transcribed spacer(rDNA-ITS) and intergenic Bloc region fragments(Bloc) of the fungus Bbr4941, and the bioassay against the adult of S. citri were carried out. The results showed that the entomopathogenic fungus was identified as Beauveria brongniartii. When the adults of S. citri were dealt with Bbr4941 suspension(5.0×10~7 spore·mL~(-1)), the corrected accumulative mortality was 100% and LT_(50) value was 3.916 d after 7 days. Our results showed that Bbr4941 had great potential for the biocontrol of S. citri.
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,2004:238-239.
[2]蔡明段,易干军,彭成绩.柑橘病虫害原色图鉴[M].北京:中国农业出版社,2011:197-198.
[3]王龙江,陆家逸,毛润乾.有机金桔园柑桔灰象甲调查初报[J].中国南方果树,2009,38(6):59-60.
[4]张小亚,陈国庆,黄振东,等.柑橘灰象甲的生物学特性及防治措施[J].浙江柑橘,2011,28(3):21-22.
[5]洪若豪.福建柑桔大害虫——灰鳞象鼻虫[J].昆虫知识,1960,6(1):16-17.
[6]洪若豪.福建柑橘大灰象甲(Sympiezomia lewisi Roel.)生活习性及防治初步研究[J].植物保护学报,1965,4(2):143-148.
[7]王旗针,杨小弟.柑桔灰象甲的生活习性及防治[J].江西园艺,2002,4:10.
[8]余舜尧,汪荣灶.茶树食叶象虫的发生与控制措施[J].江西农业科技,2004,6:30.
[9]毕旭灿,方泉寿,徐有滨,等.常山胡柚灰象甲的发生与防治对策[J].浙江柑橘,2006,23(3):30-32.
[10]吕利华,冯明光.布氏白僵菌昆虫寄主名录[J].中国生物防治,1995,3:144-144.
[11]农向群.布氏白僵菌的研究与应用[J].植物保护学报,2000,27(1):84-87.
[12]Zimmermann G.Review on safety of the entomopathogenic fungi Beauveria bassiana and Beauveria brongniartii[J].Biocontrol Science and Technology,2007,17(6):553-596.
[13]苏品,廖晓兰,张亚,等.白僵菌防治花生地蛴螬研究综述[J].广西农业科学,2009,40(4):373-377.
[14]林华峰,王萍莉,张磊,等.布氏白僵菌和金龟子绿僵菌两变种的生长性状及其对蛴螬的毒力测定[J].中国生物防治,2006,22(2):123-127.
[15]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.
[16]White TJ,Bruns T,Lee S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[M]//Innis MA,Gelfand DH,Sninsky JJ,et al.PCR protocols:a guide to methods and applications.San Diego:Academic Press,1990:315-322.
[17]Rehner SA,Posada F,Buckley EP,et al.Phylogenetic origins of African and Neotropical Beauveria bassiana s.l.pathogens of the coffee berry borer,Hypothenemus hampei[J].Journal of Invertebrate Pathology,2006,93(1):11-21.
[18]Tamura K,Dudley J,Nei M,et al.MEGA4:molecular evolutionary genetics analysis(MEGA)software version4.0[J].Molecular Biology and Evolution,2007,24:1596-1599.
[19]王定锋,曾明森,王庆森,等.球孢白僵菌不同分离株生物学特性及对茶丽纹象甲成虫的杀虫活性研究[J].福建农业学报,2012,27(7):750-754.
[20]王定锋,李良德,黎健龙,等.茶角胸叶甲高毒力球孢白僵菌菌株的筛选[J].茶叶科学,2017,37(3):229-236.
[21]Clarkson JM,Charnley AK.New insights into the mechanisms of fungal pathogenesis in insects[J].Trends in Microbiology,1996,4(5):197-203.
[22]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.
[23]Johny S,Kyei-Poku G,Gauthier D,et al.Characterization and virulence of Beauveria spp.recovered from emerald ash borer in southwestern Ontario,Canada[J].Journal of Invertebrate Pathology,2012,111:41-49.
[24]Carrillo-Benítez MG,Guzmán-Franco AW,Alatorre-Rosas R,et al.Diversity and genetic population structure of fungal pathogens infecting white grub larvae in agricultural soils[J].Microbial ecology,2013,65:437-449.
[25]Meyling NV,Pilz C,Keller S,et al.Diversity of Beauveria spp.isolates from pollen beetles Meligethes aeneus in Switzerland[J].Journal of Invertebrate Pathology,2012,109(1):76-82.
[26]Medo J,Michalko J,MedováJ,et al.Phylogenetic structure and habitat associations of Beauveria species isolated from soils in Slovakia[J].Journal of Invertebrate Pathology,2016,140:46-50.
[2]蔡明段,易干军,彭成绩.柑橘病虫害原色图鉴[M].北京:中国农业出版社,2011:197-198.
[3]王龙江,陆家逸,毛润乾.有机金桔园柑桔灰象甲调查初报[J].中国南方果树,2009,38(6):59-60.
[4]张小亚,陈国庆,黄振东,等.柑橘灰象甲的生物学特性及防治措施[J].浙江柑橘,2011,28(3):21-22.
[5]洪若豪.福建柑桔大害虫——灰鳞象鼻虫[J].昆虫知识,1960,6(1):16-17.
[6]洪若豪.福建柑橘大灰象甲(Sympiezomia lewisi Roel.)生活习性及防治初步研究[J].植物保护学报,1965,4(2):143-148.
[7]王旗针,杨小弟.柑桔灰象甲的生活习性及防治[J].江西园艺,2002,4:10.
[8]余舜尧,汪荣灶.茶树食叶象虫的发生与控制措施[J].江西农业科技,2004,6:30.
[9]毕旭灿,方泉寿,徐有滨,等.常山胡柚灰象甲的发生与防治对策[J].浙江柑橘,2006,23(3):30-32.
[10]吕利华,冯明光.布氏白僵菌昆虫寄主名录[J].中国生物防治,1995,3:144-144.
[11]农向群.布氏白僵菌的研究与应用[J].植物保护学报,2000,27(1):84-87.
[12]Zimmermann G.Review on safety of the entomopathogenic fungi Beauveria bassiana and Beauveria brongniartii[J].Biocontrol Science and Technology,2007,17(6):553-596.
[13]苏品,廖晓兰,张亚,等.白僵菌防治花生地蛴螬研究综述[J].广西农业科学,2009,40(4):373-377.
[14]林华峰,王萍莉,张磊,等.布氏白僵菌和金龟子绿僵菌两变种的生长性状及其对蛴螬的毒力测定[J].中国生物防治,2006,22(2):123-127.
[15]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.
[16]White TJ,Bruns T,Lee S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[M]//Innis MA,Gelfand DH,Sninsky JJ,et al.PCR protocols:a guide to methods and applications.San Diego:Academic Press,1990:315-322.
[17]Rehner SA,Posada F,Buckley EP,et al.Phylogenetic origins of African and Neotropical Beauveria bassiana s.l.pathogens of the coffee berry borer,Hypothenemus hampei[J].Journal of Invertebrate Pathology,2006,93(1):11-21.
[18]Tamura K,Dudley J,Nei M,et al.MEGA4:molecular evolutionary genetics analysis(MEGA)software version4.0[J].Molecular Biology and Evolution,2007,24:1596-1599.
[19]王定锋,曾明森,王庆森,等.球孢白僵菌不同分离株生物学特性及对茶丽纹象甲成虫的杀虫活性研究[J].福建农业学报,2012,27(7):750-754.
[20]王定锋,李良德,黎健龙,等.茶角胸叶甲高毒力球孢白僵菌菌株的筛选[J].茶叶科学,2017,37(3):229-236.
[21]Clarkson JM,Charnley AK.New insights into the mechanisms of fungal pathogenesis in insects[J].Trends in Microbiology,1996,4(5):197-203.
[22]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.
[23]Johny S,Kyei-Poku G,Gauthier D,et al.Characterization and virulence of Beauveria spp.recovered from emerald ash borer in southwestern Ontario,Canada[J].Journal of Invertebrate Pathology,2012,111:41-49.
[24]Carrillo-Benítez MG,Guzmán-Franco AW,Alatorre-Rosas R,et al.Diversity and genetic population structure of fungal pathogens infecting white grub larvae in agricultural soils[J].Microbial ecology,2013,65:437-449.
[25]Meyling NV,Pilz C,Keller S,et al.Diversity of Beauveria spp.isolates from pollen beetles Meligethes aeneus in Switzerland[J].Journal of Invertebrate Pathology,2012,109(1):76-82.
[26]Medo J,Michalko J,MedováJ,et al.Phylogenetic structure and habitat associations of Beauveria species isolated from soils in Slovakia[J].Journal of Invertebrate Pathology,2016,140:46-50.