Resumo

ALVAREZ-YEPES, Daniela; GUTIERREZ-SANCHEZ, Pablo A  e  MARIN-MONTOYA, Mauricio. Genome sequencing of Potato yellow vein virus (PYVV) and development of a molecular test for its detection. Bioagro [online]. 2017, vol.29, n.1, pp.3-14. ISSN 1316-3361.

Potato yellow vein virus (PYVV) is one of the most limiting viruses of potato crops in the Andean region. This virus is transmitted by whiteflies and infected tuber-seeds making the availability of adequate detection tools a very important need for management of the disease. In this study, the genome sequence of a PYVV isolate was obtained using the next generation sequencing (NGS) of infected leaves from a Solanum phureja plant in La Unión (Antioquia, Colombia) and, based on the genome data, three primer pairs for PYVV detection by RT-PCR and RT-qPCR were designed and tested in asymptomatic (12) and symptomatic (4) samples. The PYVV genome comprised three RNA segments of 8032, 5330 and 3891 nt containing a total of ten open reading frames. Comparison with the only PYVV genome sequence (PRJNA14924) available revealed an identity above 99.2 %. Primers targeting the coat (CP) sequence detected PYVV by RT-qPCR in five asymptomatic leaf samples (Ct=16.55-29.34; Tm=77.3-77.8 °C) and in the four samples showing vein-yellowing symptoms. Detection by RT-PCR resulted in fragments of the expected size for both the CP (496 bp) and the minor coat (793 bp) in symptomatic leaves. No amplification products were observed in asymptomatic material using these primers. We propose the use of RT-qPCR in tuber-seed certification programs and RT-PCR in phylogenetic and molecular variability studies of PYVV.

Palavras-chave : Crinivirus; deep sequencing; NGS; real-time RT-PCR; Solanum phureja.

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