Abstract

MUNOZ ESCUDERO, Daniel; GUTIERREZ SANCHEZ, Pablo Andrés  and  MARIN MONTOYA, Mauricio. Detection and molecular characterization of Potato virus Y (PVY) in potato (Solanum tuberosum L.) crops in northern Antioquia, Colombia. Rev. Protección Veg. [online]. 2016, vol.31, n.1, pp. 9-19. ISSN 2224-4697.

The aim of this work was to evaluate the techniques TAS-ELISA and IC-RT-qPCR as detection tools for Potato virus Y (PVY) in foliar tissues of potato (Solanum tuberosum L.). Leaf samples of the potato variety Diacol-Capiro from three different plots in northern Antioquia (Colombia) were used. Additionally, the complete genome sequence of this virus was obtained using Next-generation sequencing (NGS). PVY detection using both techniques differed significantly in sensitivity, as 88.8% of the samples tested positive for PVY using IC-RT-qPCR in contrast to only 33.3% with TAS-ELISA. Sanger sequencing of the RT-qPCR amplicons and the comparison with the melting temperatures of the positive control (77.5°C±1°C) were used to confirm the presence of PVY. Bioinformatics analysis resulted in a contig of 9701 nt with an average sequence depth of 716x. The PVY_Yarumal contains an ORF coding for a 3061 residue protein flanked by 5' and 3' untranslated regions of 187 and 328 nt, respectively. Phylogenetic analysis using the complete polyprotein CDS and CP regions suggests phylogenetic identity with necrotic strain PVY^NTN; sequence motifs K₄₀₀E₄₁₉and QELA typical of PVY^NTN were also found in PVY_Yarumal. These results suggest that urgent measures are required to strengthen current integrated virus disease management in the potato agroindustry in Colombia by using highly sensitive detection techniques based on real-time RT-PCR.

Keywords : ELISA; NGS; Potyvirus; RT-qPCR; Solanaceae.

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