Potato virus Y (PVY) detection in a single aphid by one‐step RT‐PCR with boiling technique

Abstract

Potato virus Y (PVY) (Potyviridae: potyvirus) is a serious emerging virus affecting seed potato worldwide. It affects the seed potato by transmitting non‐persistently via aphids. Sometimes this virus induces symptomless infection and is hard to detect in potato. So, it requires a specific and quick diagnosis for efficient examination. Recently, a reverse‐transcription polymerase chain reaction (RT‐PCR)‐based PVY detection method has been developed from plant as well as insect vectors. However, it is a complex and time consuming method. Here, we developed a simple PVY detection method that uses boiling for releasing the viral RNA from aphid stylets, and amplification by PVY‐specific primers located in the viral coat protein gene. The method is suitable for various strains. This simplified method could save time compared to earlier detection methods because of the simplified RNA extraction step. Following this procedure, we tested this one‐step RT‐PCR‐based PVY detection method by using three PVY vectoring aphid species (Myzus persicae, Aphis gossypii and Macrosiphum euphorbiae) as well as other sucking insects such as thrips, Frankliniella occidentalis. The reliability of a newly developed primer set was suitable for RT‐PCR and procedures were successfully demonstrated for virus detection. This PVY detection method is rapid, easy to use and suitable for large‐scale testing in laboratories of seed potato, and could potentially be applied to virus‐free seed potato production.

A plant virus satellite RNA directly accelerates wing formation in its insect vector for spread

“…Quantitative PCR was performed as described above. The partial ribosomal protein S3 ( Rps3 ) gene from M. persicae was used as the reference gene and amplified by primer pair 5′-CGAGCTGCTCCATCTCGTA-3′/5′-CCCACTTTCCATGATGAATCTCA-3′, which was designed by Kim et al 31 . The primer pair for CMV was described above.…”

Molecular Evolutionary Analysis of Potato Virus Y Infecting Potato Based on the VPg Gene

“…As one of the top 10 most famous plant viruses (Scholthof et al, 2011), some aspects of the plant pathology, molecular biology and population genetics of PVY have been studied intensively (Lorenzen et al, 2006; Chikh-Ali et al, 2013; Kim et al, 2016; Gao et al, 2017; Bai et al, 2019). However, few studies have focused on the evolutionary dynamics and timescale of PVY.…”

Population dynamics of aphid species in Korean seed potato cultivation area over four decades

“…Previously our research group attempted to establish a more comprehensive control program for aphids including virus and insecticide resistance management in seed potato cultivation (Kim et al . , ). Here we note one of the important case studies in aphid population and migration changes during some decades in Korea.…”