New qPCR-based system for detection of potato viruses PVY and PLRV in aphid vectors

Abstract

The most important potato viruses, PVY and PLRV, are transmitted by vector aphids, particularly Myzus persicae under the climatic conditions of Czechia. The integrated seed-potato protection requires detailed information on virus transmission potential in the potato fields. We developed a new system for simultaneous detection of PVY and PLRV in aphid vectors using two pairs of virus-specific primers targeting conserved genomic regions. The primers function reliably in duplex SYBR Green-based one-step qPCR combined with melting analysis, enabling virus identification by distinct amplicon melting temperatures. The method was validated using diverse virus strains from gene bank collections, artificially infected aphids and aphids collected during field monitoring. The presence of PLRV and PVY was assessed in 446 aphids, including Myzus persicae , Aphis nasturtii and Phorodon humuli , captured using yellow pan traps during 2024. Of these, 42% were PVY-positive, 2.6% PLRV-positive and 1.2% carried mixed infection. The frequency of PVY- and PLRV-positive aphids corresponded with viruses occurrence in postharvest tubers testing. This methodology is suitable for detection of PVY and PLRV in potato plants, rapid evaluation of aphid infection potential in potato fields and studies of viruses transmissibility by other aphid species.

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