华北大黑鳃金龟致病球孢白僵菌JCF的鉴定和致病力测定  

Identification and pathogenicity determination of Beauveria bassiana strain JCF from Holotrichia oblita(Coleoptera:Scarabaeidae)

作　　者：曹伟平[1] 程佳旭 丰硕 赵聚莹[1] 陈丹[1] 宋健[1] CAO Wei-Ping;CHENG Jia-Xu;FENG Shuo;ZHAO Ju-Ying;CHEN Dan;SONG Jian(IPM Center of Hebei Province,Key Laboratory of Integrated Pest Management on Crops in Northern Region of North China,Ministry of Agriculture,Plant Protection Institute,Hebei Academy of Agricultural and Forestry Sciences,Baoding 071000,China)

机构地区：[1]河北省农林科学院植物保护研究所,河北省农业有害生物综合防治工程技术研究中心,农业部华北北部作物有害生物综合治理重点实验室,保定071000

出　　处：《昆虫学报》2023年第8期999-1008,共10页Acta Entomologica Sinica

基　　金：河北省农林科学院科技创新专项课题(2022KJCXZX-ZBS-6);河北省重点研发计划项目(21326505D);河北省油料产业技术体系植保与土肥控制岗(HBCT2023030206);国家植物保护保定观测试验站项目(NAES087PP14)。

摘　　要：【目的】华北大黑鳃金龟Holotrichia oblita是花生等多种农作物、果林及苗圃的重要害虫,长期以来主要以化学防治为主。本研究测定了球孢白僵菌Beauveria bassiana菌株JCF对华北大黑鳃金龟幼虫和成虫的致病力,为华北大黑鳃金龟的生物防治提供真菌资源。【方法】对罹患病害的华北大黑鳃金龟虫尸上的寄生真菌进行分离及回接试验,分离纯化出致病菌株JCF;利用光学显微镜观察菌株形态学特征,PCR扩增JCF的rDNA-ITS序列并测序,构建JCF系统发育树对菌株进行分子鉴定;采用浸渍法和田间模拟试验测定其对华北大黑鳃金龟成虫和幼虫的致病力。【结果】JCF菌株菌落初期为白色绒状,后期变为黄色或浅黄色絮状,背面无色。产孢细胞呈瓶状簇生于分生孢子梗上,分生孢子梗呈穗状,着生在营养菌丝上;分生孢子单生,透明,近球形,直径(2.0~3.5)μm×(1.5~2.0)μm。rDNA-ITS序列分子鉴定表明菌株JCF为球孢白僵菌。这一球孢白僵菌菌株JCF对华北大黑鳃金龟成虫的致死中浓度(medium lethal concentration,LC_(50))值为2.124×10^(7)孢子/mL,用其2×10^(8),1×10^(8)和5×10^(7)孢子/mL孢悬液处理华北大黑鳃金龟成虫时,致死中时(medium lethal time,LT_(50))值分别为6.267,6.852和7.455 d;球孢白僵菌JCF菌株对华北大黑鳃金龟1,2和3龄幼虫的LC_(50)值分别为5.47×10^(6),1.59×10^(8),2.50×10^(8)孢子/mL;1×10^(8)孢子/mL浓度下,球孢白僵菌JCF菌株对华北大黑鳃金龟1和2龄幼虫的LT_(50)值分别为8.6和18.6 d;5×10^(8)孢子/mL浓度下,球孢白僵菌JCF菌株对华北大黑鳃金龟2和3龄幼虫的LT_(50)值分别为10.3和13.2 d。田间模拟试验2×10^(8)孢子/mL球孢白僵菌JCF孢悬液对华北大黑鳃金龟成虫和幼虫同样表现出良好的致病效果。【结论】球孢白僵菌菌株JCF对华北大黑鳃金龟成虫和幼虫均有较高致病力,为开发白僵菌制剂防治金龟科害虫提供了新的菌种资源,对生�【Aim】Holotrichia oblita is an important insect pest of various crops such as peanuts,fruit forests and nurseries,and it has been mainly controlled by chemical control for a long time.In this study,the pathogenicity of Beauveria bassiana strain JCF to the larvae and adults of H.oblita was determined to provide fungal resources for the biological control of H.oblita.【Methods】The parasitic fungi on the dead body of the diseased H.oblita were isolated and rejoined,and the pathogenic strain JCF was isolated and purified.The morphological characteristics of the strain were observed by light microscope.The rDNA-ITS sequence of JCF was amplified by PCR and then sequenced,and the phylogenetic tree of JCF was constructed for molecular identification.The virulence of the JCF strain to the adults and larvae of H.oblita was determined by immersion method and field simulation test.【Results】The colony of the strain JCF with colorless back is white fluffy at the initial stage,then becomes yellow or light yellow flocculent at the late stage.The conidiogenous cells are clustered on the conidiophore in a bottle shape,and the conidiophore is spike-shaped and attached to the vegetative hypha.Conidia are solitary,transparent,subglobose,and(2.0-3.5)μm×(1.5-2.0)μm in diameter.The molecular identification of rDNA-ITS sequence showed that the strain JCF was B.bassiana.The medium lethal concentration(LC_(50))value of B.bassiana strain JCF against H.oblita adults was 2.124×10^(7)conidia/mL,and the medium lethal time(LT_(50))values of this strain at the concentrations of 2×10^(8),1×10^(8)and 5×10^(7)conidia/mL against H.oblita adults were 6.267,6.852 and 7.455 d,respectively.The LC_(50)values of B.bassiana strain JCF against the 1st,2nd and 3rd instar larvae of H.oblita were 5.47×10^(6),1.59×10^(8)and 2.50×10^(8)conidia/mL,respectively.The LT_(50)values of B.bassiana strain JCF at the concentration of 1×10^(8)conidia/mL against the 1st and 2nd instar larvae of H.oblita were 8.6 and 18.6 d,respectively,and those of B.ba

关 键 词：球孢白僵菌 华北大黑鳃金龟 致病力 RDNA-ITS序列分析 分离鉴定 

分 类 号：Q965.8[生物学—昆虫学]