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Acta Biológica Colombiana
Print version ISSN 0120-548X
Abstract
GARCIA RUIZ, Daniela; GUTIERREZ SANCHEZ, Pablo and MARIN MONTOYA, Mauricio. Phylogenetic Analysis and Molecular Variability of Potato virus X (PVX) in Potato Crops of Antioquia. Acta biol.Colomb. [online]. 2016, vol.21, n.1, pp.111-122. ISSN 0120-548X. https://doi.org/10.15446/abc.v21n1.51398.
In this study, the phylogenetic relationships and molecular variability of PVX isolates from leaf samples of Solanum tuberosum subsp. andigena var. Diacol-Capiro and S. phureja var. Criolla Colombia in Antioquia were analyzed. Sequences were obtained using Next Generation Sequencing (NGS) of bulk samples and Sanger sequencing. DAS-ELISA (Agdia-PSA10000) revealed infection levels of 14.7 % and 13.3 % leaf samples of Diacol-Capiro and Criolla Colombia, respectively. The presence of PVX was further confirmed by IC-RT-qPCR in eight samples, which resulted in Ct values in the 15.04-27.59 range and two melting temperatures (Tm1 = 80.3 °C ± 0.5 and Tm2 = 83.3 °C ± 0.5). These results suggest the presence of at least two PVX variants in Antioquia. Using NGS, PVX was detected at low levels in leaf samples of Criolla Colombia, which resulted in contigs for most ORFs of the viral genome; NGS did not detect PVX in Diacol-Capiro samples. Phylogenetic analysis using capsid and replicase sequences separated PVX isolates into two groups within the Eurasian class (I). The high levels of PVX infection detected in potato crops in Antioquia and the presence of at least two variants highlight the need to strenghten current tuber seed certification programs aimed at controlling the spread of this virus.
Keywords : DAS-ELISA; NGS; Potexvirus; RT-qPCR; Solanum spp; variants.