A novel virus that infecting hypovirulent strain XG36-1 of plant fungal pathogen Sclerotinia sclerotiorum

Sclerotinia sclerotiorum is a notorious plant fungal pathogen which spreads across the world. Hypovirulence is a phenomenon where the virulence of fungal pathogens is decreased, even lost, due to mycovirus infection. The potential of hypoviruses for biological control of the chestnut blight fungus (Cryphonectria parasitica) has attracted much interest, and has led to discovery of new hypovirulent strains in other fungi.

A hypovirulent strain, strain XG36-1, was isolated from a typical lesion on the stem of rapeseed (Brassica napus) caused by Sclerotinia sclerotiorum. Strain XG36-1 grew on PDA very slowly (average 2.5 ± 0.1 mm/d) with sectoring, and developed abnormal colony morphology with few sclerotia. Unlike health strains (such as wildtype strain XG-13), it was unable to induce lesions on detached leaves of rapeseed. Sclerotia of strain XG36-1 produced apothecia rarely. A sexual progeny test showed that the phenotypes of all 104 sexual progeny were not different from wildtype strain XG-13 which shows normal phenotype of S. sclerotiorum, and protoplast regeneration tests showed that 25.5% of the regenerants of strain XG36-1 were recovered fully. Furthermore, the hypovirulence and its associated traits could be transmitted to XG36-1A34 ^R , a hygromycin-resistance gene labelled sexual progeny of strain XG36-1, by hyphal anastomosis. Transmission electron microscope (TEM) observation showed that the cytoplasm of strain XG36-1 was destroyed and granulated; the membranes of nuclei and mitochondria were disintegrated; and mitochondrial cristae were cavitated. Viral particles (about 40 nm) in hyphae of strain XG36-1, but not in its sexual progeny and wildtype strain XG-13, could be observed with TEM, and several virus-like particles were uniquely enveloped by single layer membrane in the cells of strain XG36-1. Furthermore, the viral particles could be co-transmitted with the hypovirulence traits through hyphal anastomosis.