Bashir Nemat Sokhandan, Zarghani Shaheen Nourinejhad, Hejazi Mohammad Saeid
Plant Protection Department, University of Tabriz, Iran.
Virus Res. 2007 Sep;128(1-2):144-8. doi: 10.1016/j.virusres.2007.04.013. Epub 2007 May 22.
Enzyme-linked immunosorbent assay (ELISA) testing of 126 grapevine samples, from vineyards in the northwest region of Iran, detected Grapevine fanleaf virus (GFLV) in 33 samples. Total RNA from eight of the infected samples were subjected to reverse transcription polymerase chain reaction (RT-PCR) analysis using primers which corresponded to the virus coat protein and 3' non coding region of RNA 2. An expected 1620 bp DNA fragment was amplified from all the tested samples. PCR products from isolates B5, S1 and SH3 were cloned and the nucleotide sequences of three clones from each isolate were determined. The sequences showed that a DNA fragment of 1623 bp from isolate S1 and 1629bp from isolates B5 and SH3 were amplified. The fragments covered 1481 nucleotides of the 3' proximal region of the CP gene plus 142 or 148 nucleotides of the 3' non coding region. Alignment of the sequences revealed over 99% identities among clones from each isolate and 83-93% among clones from different isolates. Identities of 83-94% were found between the isolates from Iran and previously reported GFLV strains/isolates. Phylogenetic analysis based on CP sequences showed that isolates S1 and SH3 formed a distinct cluster but isolate B5 clustered with previously reported GFLV strains. This is the first report on sequence analysis of nearly full-length CP cDNA clones of GFLV isolates from Iran.
对来自伊朗西北部葡萄园的126个葡萄样本进行酶联免疫吸附测定(ELISA)检测,在33个样本中检测到葡萄扇叶病毒(GFLV)。使用与病毒外壳蛋白和RNA 2的3'非编码区对应的引物,对8个受感染样本的总RNA进行逆转录聚合酶链反应(RT-PCR)分析。从所有测试样本中扩增出预期的1620 bp DNA片段。对分离株B5、S1和SH3的PCR产物进行克隆,并测定每个分离株三个克隆的核苷酸序列。序列显示,从分离株S1扩增出1623 bp的DNA片段,从分离株B5和SH3扩增出1629 bp的DNA片段。这些片段覆盖了CP基因3'近端区域的1481个核苷酸以及3'非编码区的142或148个核苷酸。序列比对显示,每个分离株的克隆之间同一性超过99%,不同分离株的克隆之间同一性为83-93%。在来自伊朗的分离株与先前报道的GFLV株系/分离株之间发现同一性为83-94%。基于CP序列的系统发育分析表明,分离株S1和SH3形成一个独特的簇,但分离株B5与先前报道的GFLV株系聚类。这是关于来自伊朗的GFLV分离株近乎全长CP cDNA克隆序列分析的首次报道。