Relative quantitation goes viral: An RT-qPCR assay for a grapevine virus.

Abstract:

:Accurate detection and quantitation of viruses can be beneficial to plant-virus interaction studies. In this study, three SYBR green real-time RT-PCR assays were developed to quantitate grapevine leafroll-associated virus 3 (GLRaV-3) in infected vines. Three genomic regions (ORF1a, coat protein and 3'UTR) were targeted to quantitate GLRaV-3 relative to three stably expressed reference genes (actin, GAPDH and α-tubulin). These assays were able to detect all known variant groups of GLRaV-3, including the divergent group VI, with equal efficiency. No link could be established between the concentration ratios of the different genomic regions and subgenomic RNA (sgRNA) expression. However, a significant lower virus concentration ratio for plants infected with variant group VI compared to variant group II was observed for the ORF1a, coat protein and the 3'UTR. Significant higher accumulation of the virus in the growth tip was also detected for both variant groups. The quantitation of viral genomic regions under different conditions can contribute to elucidating disease aetiology and enhance knowledge about virus ecology.

journal_name

J Virol Methods

authors

Bester R,Pepler PT,Burger JT,Maree HJ

doi

10.1016/j.jviromet.2014.09.022

subject

Has Abstract

pub_date

2014-12-15 00:00:00

pages

67-75

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(14)00377-2

journal_volume

210

pub_type

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