Preparation and Application of Polyclonal Antibodies against the Coat Protein of Fungal Virus Beauveria Bassiana Chrysovirus 2(BbCV2)
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.
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