Heterologous Expression of Cryphonectria Hypovirus 1 in Fusarium verticillioides Reveals Stable Replication and Reduced Fumonisin Production

Abstract

Mycoviruses represent promising tools for the biological control of phytopathogenic fungi. However, their application is often constrained by host specificity and low natural prevalence in certain pathogens. Fusarium verticillioides, a major maize pathogen and fumonisin B1 (FB1) producer, harbors very few known mycoviruses, a circumstance that limits the development of virus-based control strategies. In this study, we explored a virocontrol approach for F. verticillioides based on the use of a heterologous mycovirus, Cryphonectria hypovirus 1 (CHV1). For the artificial transfection of F. verticillioides with CHV1, we used an infectious cDNA clone of the virus and protoplast-mediated transformation. Through RT-PCR and double-stranded RNA (dsRNA) purification by chromatography on cellulose, it was confirmed that the CHV1 infectious cDNA clone is stably integrated into the fungal genome, giving rise to autonomous virus replication in the cytoplasm of F. verticillioides cells. Characterization of four independent transformants harboring CHV1 showed a significant reduction in FB1 production compared to the parental uninfected strain and only moderate alterations in vegetative growth. The CHV1-associated reduction in FB1 was confirmed in maize grown under both greenhouse and field conditions. Our findings provide evidence that heterologous mycoviruses can modulate mycotoxin production in F. verticillioides, thus expanding the pool of viral species that can be explored as potential biocontrol agents in this pathogen. Further work will be required to elucidate the underlying mechanisms of CHV1-associated downregulation of FB1 production in F. verticillioides and assess its potential application in virocontrol strategies.