一种用于检测茄科植物中马铃薯纺锤块茎类病毒的简化斑点杂交协议。

A Simplified Dot-Blot Hybridization Protocol for Potato spindle tuber viroid Detection in Solanaceae.

作者信息

Kekstidou Oxana, Varveri Christina, Vassilakos Nikon

机构信息

Scientific Directorate of Phytopathology, Laboratory of Virology, Benaki Phytopathological Institute, Athens, Greece.

出版信息

Methods Mol Biol. 2022;2316:89-96. doi: 10.1007/978-1-0716-1464-8_9.

DOI:10.1007/978-1-0716-1464-8_9

PMID:34845688

Abstract

A simplified dot-blot hybridization protocol for Potato spindle tuber viroid (PSTVd) detection in Solanaceae species is described here. The protocol uses an RNA DIG-labeled probe and a simplified extraction procedure that avoids the use of hazardous chemicals. PSTVd was detected in composite tomato leaf samples in a ratio of up to 1:15 of PSTVd-infected to non-infected tissue and in composite potato tuber samples in a ratio up to 1:5 of PSTVd-infected to non-infected tissue. In Brugmansia spp., PSTVd was detected solely in the standard sample extract preparation. The method is suitable for a reliable, large-scale sample screening especially where cost is a limiting factor.

摘要

本文描述了一种用于检测茄科物种中马铃薯纺锤块茎类病毒（PSTVd）的简化斑点杂交方案。该方案使用RNA地高辛标记探针和简化的提取程序，避免使用危险化学品。在复合番茄叶片样本中，PSTVd感染组织与未感染组织的比例高达1:15时可检测到PSTVd；在复合马铃薯块茎样本中，PSTVd感染组织与未感染组织的比例高达1:5时可检测到PSTVd。在曼陀罗属植物中，仅在标准样本提取物制备中检测到PSTVd。该方法适用于可靠的大规模样本筛选，尤其是在成本是限制因素的情况下。