Molecular tools for studying the interaction between Botrytis and the viruses BVX and BVF (NZ).

Understanding the nature of the relationship between viruses and their fungal hosts is critical in determining the ecological significance of mycoviruses and their potential usage as biological control agents. Two flexuous viruses belonging to the family Flexiviridae, Botrytis virus F (BVF) and Botrytis virus X (BVX), from Botrytis cinerea have previously been completely sequenced, providing the opportunity to examine their interaction with B. cinerea at molecular level. In addition, studying their impact on the virulence of Botrytis is also of great interest. In order to study the fungus/virus interaction four basic tools were developed: i) an efficient transfection protocol to introduce viruses into uninfected fungal isolates (ii) a transformation protocol to incorporate plasmid DNA into Botrytis, iii) a consistent and reliable real-time PCR detection method for BVF and BVX to study the effect of virus transfections, and iv) an immunoassay for BVX to visualize the virus distribution and movement within the mycelia and also between compatible fungal strains. These tools will enable the study of the relationship between the fungus and the mycoviruses at the cellular level.