Molecular Characterization of Phylogenetically Distinct Isolates of Grapevine fanleaf virus from Iran Based on 2A(HP) Gene

Abstract

Movement and coat protein genes from Grapevine fanleaf virus (GFLV) isolates have been characterized previously from Iran. In this study, an optimized reverse transcription polymerase chain reaction protocol was established to amplify RNA2 genomic segment corresponding to the hypothetical protein (2A(HP)). The sequence of 2A(HP) was compared with that of previously reported GFLV strains/isolates from other countries which showed 82-86% sequence identities. The 2A(HP) gene from Iran appeared to be standing distinct from other isolates of GFLV when genetic distance- or parsimony-based phylogeneitc analyses were carried out. The present study for the first time reports characterization of Iranian isolate of GFLV based on 2A(HP) gene.

Keywords: 2AHP gene; ELISA; GFLV; Hypothetical protein; Nepovirus; RT-PCR.

Fig. 1

RT-PCR amplification stretches of Grapevine fanleaf virus 2A^HP coding region by the use of the primer sets 5′NC/G0 (a) or 5′NC/GMPR1 (b). Lane 1 in both gels are loaded with 100 bp DNA ladder plus. a, lanes 2–6: ~800 bp products amplified from samples Kh6, Kh11, Kj12, B15, and Kj 18, respectively, lane 7: RT-PCR on healthy control. b, lanes 2–10: 1995 bp fragment resulting from amplifications from samples B4, B12, Ma13, Kj14, B15, Kj16, Kj17, Kj18, and J, respectively, lane 11: RT-PCR on healthy control

Fig. 2

A parsimony tree based on 622 nucleotide sequence of 5′ proximity of GFLV HP gene. The Iran HP isolates are marked. The bootstrap values are shown on the nodes. Branches with <50% bootstrap values are unresolved. The scale bar represents 10% bootstrap value and branch lengths are proportional to bootstrap values