Plant Protection Research Institute, Agricultural Research Council, Pretoria, South Africa.
Arch Virol. 2010 Sep;155(9):1463-9. doi: 10.1007/s00705-010-0714-0. Epub 2010 Jun 13.
The presence of rugose-wood-associated viruses of the genera Foveavirus and Vitivirus in the family Betaflexiviridae was investigated in various clones of own-rooted and grafted Vitis vinifera cv. Shiraz that were affected, or not, by Shiraz decline, and in rootstocks. RT nested-PCR amplification of double-stranded RNA using degenerate primers for the simultaneous detection of foveaviruses and vitiviruses (Dovas CI, Katis NI in J Virol Meth 170:99-106, 2003), cloning of DNA amplicons, SSCP analysis of clones, sequencing and computer-assisted analysis of sequences was used to characterize viral genetic variability. A total of 1,137 clones were analysed by SSCP, and, of those, 371 clones were sequenced. The results revealed that variants of five molecular groups belonging to the species Grapevine rupestris stem pitting-associated virus (GRSPaV), including highly divergent variants related to strain SY (Lima MF et al. in Arch Virol 151:1889-1894, 2006) were present in plants of various clones of Shiraz regardless of their Shiraz decline status, and in rootstocks. Grapevine virus A (GVA) and grapevine virus B (GVB) were detected in a relatively small number of plants. This study suggested no involvement of GRSPaV, GVA or GVB in Shiraz decline.
本研究采用 RT-nested-PCR 技术,利用针对 Foveavirus 和 Vitivirus 属病毒的简并引物(Dovas CI, Katis NI in J Virol Meth 170:99-106, 2003),对来自不同来源的 own-rooted 和 grafted Shiraz 葡萄品种克隆,以及砧木进行检测,以明确贝塔 Flexiviridae 科病毒中是否存在褶皱木相关病毒(rugose-wood-associated viruses)。对扩增的双链 RNA(double-stranded RNA)进行克隆、SSCP 分析、测序以及计算机辅助序列分析,以确定病毒的遗传变异性。共对 1137 个克隆进行 SSCP 分析,其中 371 个克隆进行了测序。结果显示,5 个分子群的变异株(包括与 SY 株高度分化的变异株)存在于不同来源的 Shiraz 克隆植物中,与 Shiraz 衰退的状态无关,也存在于砧木中。葡萄 A 病毒(Grapevine virus A,GVA)和葡萄 B 病毒(Grapevine virus B,GVB)的检出率较低。本研究表明,褶皱木相关病毒、葡萄 A 病毒和葡萄 B 病毒与 Shiraz 衰退无关。