Abstract
Comprehensive next generation sequencing virus detection was used to detect the whole spectrum of viruses and viroids in selected grapevines from the Czech Republic. The novel NGS approach was based on sequencing libraries of small RNA isolated from grapevine vascular tissues. Eight previously partially-characterized grapevines of diverse varieties were selected and subjected to analysis: Chardonnay, Laurot, Guzal Kara, and rootstock Kober 125AA from the Moravia wine-producing region; plus Müller-Thurgau and Pinot Noir from the Bohemia wine-producing region, both in the Czech Republic. Using next generation sequencing of small RNA, the presence of 8 viruses and 2 viroids were detected in a set of eight grapevines; therefore, confirming the high effectiveness of the technique in plant virology and producing results supporting previous data on multiple infected grapevines in Czech vineyards. Among the pathogens detected, the Grapevine rupestris vein feathering virus and Grapevine yellow speckle viroid 1 were recorded in the Czech Republic for the first time.
Highlights
The infection of plant tissues by viral pathogens can cause significant economic losses to agriculture crops [1]
Plants of various varieties and geographical origins were included in the analyzed set; primarily from Moravia, and from wine regions in Bohemia
The eight detected viruses belong to several families: Tymoviridae (Grapevine fleck virus, Grapevine rupestris vein feathering virus, Grapevine Syrah virus 1), Closteroviridae (Grapevine leafroll-associated virus 1), and Betaflexiviridae (Grapevine Pinot gris virus, Grapevine rupestris stem pittingassociated virus, Grapevine virus A, Grapevine virus B); whereas, the two viroids (Hop stunt viroid and Grapevine yellow speckle viroid 1) belong to the family Pospiviroidae
Summary
The infection of plant tissues by viral pathogens can cause significant economic losses to agriculture crops [1]. Many different viral pathogens occur, especially on vegetatively propagated crops, which have accumulated these pathogens over the centuries and millennia. Pathogenspecific detection techniques, either immunoenzymatic or nucleic acid-based methods, cannot get the overall picture of all of the pathogens present in the plants tested, where each sample must be tested for several pathogens simultaneously. When more pathogens with economical impacts occur in a tested crop in a given region, and needs to be tested for, such a procedure starts to be laborious and inconvenient. This is the case with grapevines, which host about 70. PLOS ONE | DOI:10.1371/journal.pone.0167966 December 13, 2016
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