A simple and rapid method for processing tissue infected with plum pox potyvirus for use with specific 3’non‐coding region RT‐PCR assays1

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Abstract
A rapid, simple method for preparing plant tissue infected with plum pox potyvirus (PPV) using a commercial product known as Gene Releaser is described. The Gene Releaser polymeric matrix method produces plant extracts suitable for PCR amplification without the use of organic solvents, ethanol precipitation, or additional nucleic acid purification techniques. We also describe the development of a PPV‐specific amplification assay based on the unique 220 nucleotides present in the 3’non‐coding region of the PPV genome. This paper demonstrates the simplicity of the Gene Releaser method combined with the accuracy of the PCR assay for the detection of multiple PPV strains from Spain, France, Greece, Italy, Germany, Egypt, Hungary, and Romania. Amplification of the 3’non‐coding region of potato Y potyvirus (PVY) using primers for the 3’non‐coding region of the PVY genome was also possible with Gene Releaser preparation of viruliferous Myzus persicae to demonstrate the potential usefulness of this work for PPV detection from aphids.