IDENTIFICATION AND DETECTION OF IRANIAN ISOLATES OF Grapevine fanleaf virus USING GREEN-GRAFTING AND RT-PCR

Grapevine fanleaf virus (GFLV) is one of the most destructive grapevine viruses worldwide. The best strategy to control the virus is to cultivate virus free plant material in non-infested soils. Therefore, detection of the virus plays a key role in control of GFLV. To establish a reliable and sensitive technique for detection of GFLV in the stock grapevines or in preparation of virus free grapevines, samples from different vineyards of Iran were collected in 2009 and their infection by GFLV was assessed by DAS-ELISA. Fourteen ELISA positive samples were subjected to RT-PCR using GFLV published primers and the primers designed in this study. Most of the primers available in literature failed to detect GFLV in all 14 tested samples, whereas the primers designed in this study detected the virus in all mentioned samples. The efficacy of the designed primers in this study was confirmed by detection of the virus in further 35 ELISA positive samples collected from other provinces in 2010. Moreover, GFLV infectivity of grapevines for the first time was confirmed by green grafting. The results revealed that the combination of the green-grafting with RT-PCR method, using the introduced primers in this study, offers a reliable method for detection of GFLV isolates.