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چکیده
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A wine sample of Cabernet Sauvignon was obtained from the Australian Wine Research Institute (Adelaide, South Australia). This wine was made in 2004 by a Victorian winery (Yarra Glen, Australia). Total nucleic acids (TNA) were extracted from the wine sample as described before (Habili et al., 2012). For this purpose, five ml wine batches were dialysed to dryness against solid PVP 40 and resuspended in Lysis buffer of McKenzie et al (1997). Viroid amplicons obtained from single tube RT-PCR and the products were cloned using the pGEM-T vector (Promega) system and sequenced by AGRF (Adelaide). BLASTn analysis showed that the complete sequence of GYSVd-1detected in a Cabernet Sauvignon wine had 99.4% identity with a variant of this viroid reported from Yazd (Iran). Its predicted secondary structure and its five functional domains are indicated in Fig. 1. The accession number for the Iranian variant is KF916042. There were only two nucleotide substitutions; A234->G and the other one was a synonymous substitution of A362->U. Full length genome of HSVd was also sequenced and matched 98% to Isolate H1 from grapevine in China. The sequencing of the full length genome of AGVd detected in the same wine sample showed 99% identity with the Meyme isolate from Iran (Acc. # KF876034). GYSVd-2 was not detected using the primers given in Table 1. Attempts to detect ASSVd in any wine sample failed and our previous results (Habili et al., 2012) could not be confirmed. Detection of full-length sequence of viroids, e.g. HSVd in wines could indicate the potential dissipation of these infectious agents into a new environment.
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