Ciencia E Tecnica Vitivinicola, vol.32, no.8, pp.247-257, 2017 (SCI-Expanded)
This study has been performed on the detection and determination of Citrus Tatter Leaf Virus (CTLV), which has not been studied at the molecular level in Cyprus. Therefore, this study focused on the molecular characterization of CTLV in citrus plants in Cyprus. This paper reports the sequencing and analysis of a CTLV isolate obtained in Cyprus and examines its relationship to other CTLV isolates from different parts of the world. Surveys were conducted in various regions of Cyprus between 2013 and 2016. As a result, 84 different citrus orchards, which showed CTLV symptoms, like chlorosis, leaf wrinkled, asymmetrical leaves, yellow lesions and mosaic shaped spotswere selected for sampling. ReverseTranscriptase – Polymerase Chain Reactions were performed in the thermal cycler using one pair of CTLV specific primer (TL1F: 5′-TGAAAACCTTTGCTGCCACCTCT-3′ and TL1R: 5′-TACTCTCCGAACCTGCCTCGAAA-3′), which yielded a 309 nucleotide fragment overlapping the N – terminal part of the coat protein (CP). Polymerase Chain Reaction products were analyzed by electrophoresis in a 2 % agarose at 60 volts, for one hour with TAE buffer. DNA was visualized by staining gel in ethidium bromide (0.5 μg/mL) and photographed under UV light. Six samples were found positive for CTLV. After that, Reverse Transcriptase – Polymerase Chain Reaction products were sequenced and the nucleotide sequencing analysis showed that all CTLV isolates which were obtained from Cyprusare same and they were similar to the three different CTLV isolates from China (CTLV-BJNM-2 and CTLV-QC4) and Australia (CTLV-BDZ-1). The results revealed that, this is the first report of a complete nucleotide sequence of a CTLV isolate for Cyprus and Europe.