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.