中国生物防治学报2024，Vol.40Issue(3)：583-591,9.DOI:10.16409/j.cnki.2095-039x.2023.01.049
真菌病毒Beauveria bassiana chrysovirus 2(BbCV2)外壳蛋白多克隆抗体制备及应用
Preparation and Application of Polyclonal Antibodies against the Coat Protein of Fungal Virus Beauveria Bassiana Chrysovirus 2(BbCV2)
摘要
Abstract
Beauveria bassiana is a widely used entomogenous fungus in pest biocontrol.Mycoviruses can affect the pathogenicity and biological characteristics of B.bassiana.However,there are few reports on the preparation of antibodies against fungal virus-related proteins and their use for virus detection and localization.In the present study,the coat protein(CP)of a double stranded RNA(dsRNA)virus,Beauveria bassiana chrysovirus 2(BbCV2),which is epidemic and hypovirulent to host fungi B.bassiana,was expressed prokaryoticly.And polyclonal antibodies were prepared for the detection of the virus in and out of the host fungi body and location the virus in the host fungi spores using indirect immunofluorescence.The results showed that the expressed BbCV2-CP protein mainly existed in the form of inclusion body,with a relative molecular weight of approximately 85.7 kD.By indirect ELISA method,the prepared BbCV2-CP polyclonal antibody had a titer of 1:8192000.The results of Western blot showed that the prepared BbCV2-CP polyclonal antibody specifically bound to the antigen protein and recognized BbCV2 infected B.bassiana strains;The ELISA assay results showed that the virus BbCV2 replicated in the host fungus and was able to dissociate outside the host.According to the indirect immunofluorescence observation,the BbCV2 virus localized in the fungal nucleus.This study has established a serological detection system for the mycovirus infecting B.bassiana,providing materials for the study of the interaction between B.bassiana and mycovirus.关键词
球孢白僵菌/真菌病毒BbCV2/多克隆抗体/免疫荧光/亚细胞定位Key words
Beauveria bassiana/mycovirus BbCV2/polyclonal antibody/immunofluorescence/subcellular localization分类
农业科技引用本文复制引用
路杨,郭文博,隋丽,张庆贺,赵宇,张正坤,李启云..真菌病毒Beauveria bassiana chrysovirus 2(BbCV2)外壳蛋白多克隆抗体制备及应用[J].中国生物防治学报,2024,40(3):583-591,9.基金项目
吉林省科技厅中青年科技创新创业卓越人才(团队)项目(创新类)(20230508011RC) （团队）
国家自然科学基金(32271683) （32271683）
