Detection of five viruses infecting dormant bulbs by TaqMan-based real-time RT-PCR

Abstract

Dormant bulbs may be infected by a number of viruses which are of potential economic importance for the horticultural industry. Five important viruses in bulbs were successfully detected by TaqMan-based real-time one-step RT-PCR assays: Arabis mosaic virus (ArMV), Cucumber mosaic virus (CMV), Lily symptomless virus (LSV), Tobacco rattle virus (TRV) and Tulip virus X (TVX). Nucleic acid of appropriate quality and quantity for RT-PCR was extracted from nine bulb species using the Qiagen RNeasy plant mini kit. Depending on the assay, the sensitivity of the real-time RT-PCRs varied from being the same sensitivity to 1,000 times more sensitive than the respective conventional RT-PCRs. The reliability of the TVX real-time RT-PCR was further assessed against conventional RT-PCR using leaves and bulbs of 24 Tulipa × hybrida plants. TVX was detected in twice the number of leaf samples than bulb samples for both conventional and real-time RT-PCRs.