Kenyon Lawrence, Lebas B S M, Seal S E
Natural Resources Institute, The University of Greenwich at Medway, Central Avenue, Chatham Maritime, Kent ME4 4TB, UK.
Arch Virol. 2008;153(5):877-89. doi: 10.1007/s00705-008-0062-5. Epub 2008 Mar 11.
Yam (Dioscorea spp.) samples (n = 690) from seven South Pacific Islands were screened for badnavirus infection by ELISA using two antisera to African badnaviruses. Positive readings were obtained for 26.4-34.6% of samples representing both known (D. bulbifera, D. nummularia and D. pentaphylla) and unreported host species (D. alata, D. esculenta, D. rotundata and D. trifida) in this region. Total DNAs were extracted from 25 ELISA-positive plants and 4 ELISA-negative controls and subjected to PCR amplification with badnavirus-specific primers targeting the reverse transcriptase (RT)-RNaseH genes. All 29 samples yielded the expected size PCR-product for badnaviruses, which were cloned and sequenced. Phylogenetic analyses of the resulting 45 partial (500-527 bp) RT-RNaseH sequences revealed 11 new sequence groups with <79% nucleotide identity to each other or any EMBL sequence. Three sequences (two groups) were highly divergent to the other nine new South Pacific yam badnavirus groups (47.9-57.2% identity) and probably represent either new Caulimoviridae genera or endogenous pararetrovirus sequences. Some sequence groups appeared specific to particular Dioscorea host species. Four 99.9% identical RT-RNaseH sequences possessing nine amino acid deletions from D. esculenta from three islands represent a putative integrated sequence group. The distribution of sequence groups across the islands indicates that badnaviruses have spread extensively between islands and continents through infected germplasm.
使用两种针对非洲杆状DNA病毒的抗血清,通过酶联免疫吸附测定(ELISA)对来自南太平洋七个岛屿的690份山药(薯蓣属)样本进行了杆状DNA病毒感染筛查。在代表该地区已知宿主物种(如黄独、金钱薯和五叶薯)和未报告宿主物种(如翼薯、参薯、圆薯和 trifida薯)的样本中,26.4%-34.6%获得了阳性读数。从25株ELISA阳性植株和4株ELISA阴性对照中提取总DNA,并用针对逆转录酶(RT)-核糖核酸酶H基因的杆状DNA病毒特异性引物进行PCR扩增。所有29个样本均产生了杆状DNA病毒预期大小的PCR产物,对其进行克隆和测序。对所得的45个部分(500-527 bp)RT-核糖核酸酶H序列进行系统发育分析,发现了11个新的序列组,它们彼此之间或与任何EMBL序列的核苷酸同一性均<79%。三个序列(两个组)与其他九个新的南太平洋山药杆状DNA病毒组高度不同(同一性为47.9%-57.2%),可能代表新的花椰菜花叶病毒科属或内源性类逆转录病毒序列。一些序列组似乎特定于特定的薯蓣属宿主物种。来自三个岛屿的参薯中具有九个氨基酸缺失的四个同一性为99.9%的RT-核糖核酸酶H序列代表一个推定的整合序列组。序列组在各岛屿间的分布表明,杆状DNA病毒已通过受感染的种质在岛屿和大陆之间广泛传播。