Infection of Rosellinia necatrix with purified viral particles of a member of Partitiviridae (RnPV1-W8)

Abstract

Isolate W8 of the white root rot fungus, Rosellinia necatrix, harbors three dsRNA segments, L1-, L2- and M-dsRNAs, and showed an irregular colony margin, slow growth, and moderate virulence. The M-dsRNA was previously shown to be the genome of a partitivirus, RnPV1-W8. Here a transfection protocol was developed for RnPV1-W8. Protoplasts of two virus-free isolates of R. necatrix were inoculated with purified viral particles using a polyethylene glycol-mediated method. Virus infection was confirmed by electrophoresis and Northern analysis. RnPV1-W8 introduced into the new host isolates was transmissible via hyphal anastomosis. However, the infection had no effect on the morphology and virulence of infected isolates of R. necatrix. This is the first report on the transfection of a partitivirus for R. necatrix.

A Novel Victorivirus from a Phytopathogenic Fungus, Rosellinia necatrix, Is Infectious as Particles and Targeted by RNA Silencing

“…Protoplasts isolated from the R. necatrix strains W97 and W563 and the C. parasitica EP155 and ⌬dcl-2 strains were prepared and transfected with purified virus particles as described previously (11,12). The purified virus fractions were passed through Ultrafree-MC sterile centrifugal filter units (Millipore, Tokyo, Japan) and introduced into protoplasts in a PEG-calcium chloride-mediated manner.…”

Mycoviruses: future therapeutic agents of invasive fungal infections in humans?

“…Once fungi are successfully transfected, the observed infection patterns are very stable when subcultured [9,28]. Furthermore, viruses present in Aspergillus isolates are highly efficiently transmitted to asexual progeny [21].…”

“…Infection by the extracellular route could thus far only be achieved in special experimental settings using protoplasts. Applying this method, purified viral particles belonging to the dsRNA families of Partitiviridae, Totiviridae, and Reoviridae successfully infected fungal protoplasts [10,28,29].…”

“…Replicating mycoviruses are thus able to spread throughout the colony via plasma streaming [28,30] (Fig. 2).…”

Effect of Diaporthe RNA virus 1 (DRV1) on growth and pathogenicity of different Diaporthe species

“…However, it has to be noted that successful infections with mycoviruses with rigid virus particles have been achieved using mycoviruses from families Reoviridae (Hillman et al, 2004;Sasaki et al, 2007;Kanematsu et al, 2010) and Partitiviridae (Sasaki et al, 2006;Kanematsu et al, 2010). In these studies, infection was achieved by inoculating fungal protoplasts with virus particles.…”