Purification, Serology, and Some Properties of a Mechanically Transmissible Virus Associated with Green Ring Mottle Disease in Peach and Cherry

cDNA cloning and molecular characterization of cherry green ring mottle virus.

“…CGRMV was not mechanically transmitted to any of the herbaceous plants tested as determined by the absence of viral symptoms and negative RT-PCR assays which also agrees with previous research (Zagula et al, 1989). Because our early attempts to purify CGRMV virions directly from woody plant hosts were unsuccessful, it was only possible to isolate and clone the CGRMV genome using dsRNA isolated from infected plant tissue.…”

“…Phylogenetic analysis of both genes shows that CGRMV is most similar to ASPV, while CGRMV was most similar to carlaviruses by analysis of the replicase gene and most similar to potexviruses by analysis of the coat protein gene. Although its precise taxonomic classification remains uncertain, these molecular analyses clearly show that CGRMV is not a closterovirus as previously suggested (Zagula et al, 1989). The overall genomic organization and properties of specific CGRMV genes have attributes in common with ASPV, potexviruses and carlaviruses.…”

“…Virions of CGRMV were partially purified from 20 g of infected cherry leaf tissue using a part of the procedure of Zagula et al (1989) and analysed by SDS-PAGE. Electrophoresed proteins were transferred to a nitrocellulose membrane using a ' Mini trans-blot electrophoretic transfer cell ' (Bio-Rad) following the manufacturer's protocol.…”

“…Indicator hosts of CGRMV include Kwanzan and Shirofugen flowering cherry trees (P. serrulata L.) in which CGRMV produces vein necrosis and epinasty of leaves 2-3 months after graft-inoculation. A Michigan strain of CGRMV was isolated and characterized by Zagula et al (1989). The purified virus particles ranged from 1000-2000 nm in length and 5-6 nm in diameter, with a buoyant density of 1n24-1n25 g\cm$, an ssRNA of 2n5i 10' Da and coat protein of 25 kDa.…”

Purification and Characterization of Peach Mosaic Virus

“…The difficulty of purification of peach mosaic virus from peach is thought to be due to the relatively low concentration of the virus and the presence of large amounts of mucilage, a heteropolysaccharide which is known to cause serious difficulties in the isolation and preservation of viruses (26). In addition to this, proteins, polysaccharides, and the presence of phenolic substances such as tannins may inhibit virus infection when using inoculum prepared from peach tissue (18).…”