一种用于诊断田间和组织培养样品中葡萄类病毒的高灵敏度逆转录聚合酶链反应检测方法。

A high sensitivity RT-PCR assay for the diagnosis of grapevine viroids in field and tissue culture samples.

作者信息

Wan Chow Wah Y F, Symons R H

机构信息

Department of Plant Science and Cooperative Research Centre for Viticulture, Waite Institute, University of Adelaide, Glen Osmond, Australia.

出版信息

J Virol Methods. 1997 Jan;63(1-2):57-69. doi: 10.1016/s0166-0934(96)02115-5.

DOI:10.1016/s0166-0934(96)02115-5

PMID:9015276

Abstract

An RNA extraction procedure, modified from published methods, and a high sensitivity reverse transcription-polymerase chain reaction (RT-PCR) assay have been developed for the detection of the five viroids in grapevines. All five viroids have been found in the 10 different grape varieties tested so far. This assay, specially optimised for viroids in low copy number by careful selection of DNA primers, has been used in conjunction with dot blot hybridization assay for the study of viroids in vines regenerated by shoot apical meristem culture (SAMC) and fragmented shoot apex culture (FSAC). The data indicate a differential reduction of viroids, rather than viroid elimination, in the regenerated vines. Transmission of viroids via grape seeds was also observed.

摘要

我们开发了一种从已发表方法改进而来的RNA提取程序，以及一种高灵敏度逆转录聚合酶链反应（RT-PCR）检测方法，用于检测葡萄中的五种类病毒。到目前为止，在所测试的10个不同葡萄品种中均发现了这五种类病毒。该检测方法通过精心选择DNA引物，特别针对低拷贝数的类病毒进行了优化，并已与斑点杂交检测法结合使用，用于研究通过茎尖分生组织培养（SAMC）和茎尖切段培养（FSAC）再生的葡萄藤中的类病毒。数据表明，再生葡萄藤中的类病毒减少存在差异，而非类病毒消除。还观察到类病毒通过葡萄种子传播。