Molecular detection of Papaya meleira virus in the latex of Carica papaya by RT-PCR

Abstract

A RT-PCR assay was developed for early and accurate detection of Papaya meleira virus (PMeV) in the latex from infected papayas. The meleira disease is characterized by an excessive exudation of more fluidic latex from fruits, leaves and stems. This latex oxidises and gives the fruit a “sticky” texture. In the field, disease symptoms are seen almost exclusively on fruit. However, infected plants can be a source of virus for dissemination by insects. Primers specific for PMeV were designed based on nucleotide sequences of the viral dsRNA obtained using a RT-RAPD approach. When tested for RT-PCR amplification, one of these primers (C05-3′) amplified a 669-nucleotide fragment using dsRNA obtained from purified virus particles as a template. The translated sequence of this DNA fragment showed a certain degree of similarity to the amino acid sequence of RNA-dependent RNA polymerases from other dsRNA viruses. When used as the single primer in two RT-PCR kits available commercially, primer C05-3′ also amplified the DNA fragment from papaya latex of infected, but not from healthy plants. The RT-PCR-based method developed in this study could simplify early plant disease diagnosis, assist in monitoring the dissemination of the pathogen within and between fields, and assist in guiding plant disease management.