The most important advances in our understanding of the viral life cycle, such as genome replication, packaging, transmission, and host interactions, have been made via the development of viral infectious full-length clones. Here, we describe the detailed protocols for the construction of an infectious clone derived from Botrytis virus F (BVF), a mycoflexivirus infecting the plant pathogenic fungus Botrytis cinerea, the determination of the complete sequence of the cloned mycovirus, the preparation of fungal protoplasts, and the transfection of protoplasts using transcripts derived from the BVF infectious clone.
Keywords: Botrytis cinerea; Fungus; Mycovirus; Reverse genetics; Synthetic virus; Viral infectious clone; Virus.
© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.