Kekstidou Oxana, Varveri Christina, Vassilakos Nikon
Scientific Directorate of Phytopathology, Laboratory of Virology, Benaki Phytopathological Institute, Athens, Greece.
Methods Mol Biol. 2022;2316:89-96. doi: 10.1007/978-1-0716-1464-8_9.
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.
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。
