Orchid Fleck Virus Infecting Orchids in Paraguay: First Report and Use of Degenerate Primers for its Detection

Abstract

The presence of Orchid fleck virus (OFV) in Paraguay was confirmed in orchid plants collected during a survey carried out in 2013. Leaves displayed ringspot and fleck symptoms, and in infected tissues, non-enveloped, short, rodlike viral particles were observed. Partial OFV N and L genes were amplified using specific and degenerate primers, respectively; the nucleotide sequences demonstrated high identities (98%) with other OFV isolates. Degenerate primers for the L gene were designed considering conserved regions within all of the available OFV sequences and those from the closely related isolates citrus leprosis virus nuclear type (CiLV-N) and citrus necrotic spot virus. Degenerate primers were also successfully used for the detection of CiLV-N from infected citrus samples.

First report of Cymbidium mosaic virus on orchids in Paraguay

“…Amplicons of the expected size for CymMV (681 bp) were obtained with total RNA extracted from all samples. In addition, at least one plant of each genus was found to also be infected with either OFV or ORSV; two viruses previously reported in Paraguay (González-Segnana, 1989;Ramos-González et al, 2016). Three RT-PCR amplicons for CymMV, obtained from different plants of each genus (fifteen in total), were purified with PCR Clean-Up System (Promega, USA) and directly sequenced (Macrogen Inc., Korea).…”