Bostan Hidayet, Nie Xianzhou, Singh Rudra P
Department of Plant Protection, Faculty of Agriculture, Ataturk University, Erzurum 25240, Turkey.
J Virol Methods. 2004 Mar 15;116(2):189-93. doi: 10.1016/j.jviromet.2003.11.014.
A primer pair for reverse transcription-polymerase chain reaction (RT-PCR), based on the conserved sequences of the members of genus Pospiviroid was designed to yield a fragment of about 200 base pairs (bp). Since pospiviroids infect a large number of plants species and a few members of the genus Pospiviroid have been already detected in some ornamental plants, the primer pair was evaluated for its efficacy using ornamental plants. The method of return-polyacrylamide gel electrophoresis (R-PAGE) was used to determine the general presence of viroids in the test samples. Efficacy of the primer pair for members of genus Pospiviroid was demonstrated by the detection of Potato spindle tuber viroid (PSTVd) and Tomato chlorotic dwarf viroid (TCDVd) in potato, Chrysanthemum stunt viroid and Iresine viroid in Verbena and Vinca species, and Citrus exocortis viroid in Impatiens species. Specificity of the primer pair became evident, where additional viroids were detected by R-PAGE in Coleus and Magilla species, but they were not amplified by the Pospiviroid primer. This primer pair would be of benefit in indexing ornamental plants in quarantine samples or in viroid-free certification schemes, irrespective of their actual identity.
基于马铃薯纺锤块茎类病毒属成员的保守序列，设计了一对用于逆转录聚合酶链反应（RT-PCR）的引物，以扩增出一段约200个碱基对（bp）的片段。由于马铃薯纺锤块茎类病毒可感染大量植物物种，且已在一些观赏植物中检测到该属的少数成员，因此使用观赏植物对该引物对的有效性进行了评估。采用反向聚丙烯酰胺凝胶电泳（R-PAGE）方法来确定测试样品中类病毒的总体存在情况。通过在马铃薯中检测到马铃薯纺锤块茎类病毒（PSTVd）和番茄褪绿矮缩类病毒（TCDVd）、在马鞭草和长春花属物种中检测到菊花矮化类病毒和紫锦草类病毒、在凤仙花属物种中检测到柑橘裂皮类病毒，证明了该引物对对于马铃薯纺锤块茎类病毒属成员的有效性。引物对的特异性很明显，在彩叶草和玛吉拉属物种中通过R-PAGE检测到了其他类病毒，但它们未被马铃薯纺锤块茎类病毒引物扩增。无论观赏植物的实际身份如何，该引物对都将有助于对检疫样品中的观赏植物进行检测或用于无类病毒认证方案。
