A simple and rapid method for the detection of potato spindle tuber viroid (PSTVd) by RT-PCR
A simple and rapid method for the detection of potato spindle tuber viroid (PSTVd) by RT-PCR
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一种简单快速的 RT-PCR 检测马铃薯纺锤块茎类病毒 (PSTVd) 的方法
DOI：
10.1007/bf02360256
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发表时间：
1998
期刊：
影响因子：
2.9
通讯作者：
U. Buchta
中科院分区：
文献类型：
--
作者：
H. Weidemann;U. Buchta
描述了一种PSTVd的测试程序，其基础是将植物汁液固定在滤纸上，通过打点或组织打印，然后进行RT-PCR。试验使用了主要和第二次感染的马铃薯植株，主要是体外感染的植株，以及马铃薯块茎。印迹聚合酶链式反应适用于大样本马铃薯植株的检测，而斑点聚合酶链式反应适用于试管苗和块茎组织的检测。将一个受感染的植株组合成4或9个健康植株，对继发感染的马铃薯植株可以得到可靠的结果，但有时检测不出初发感染的试管苗中的PSTVd。打点和打印的纸方块可以在4°C的Triton X-100溶液中或干燥条件下保存至少2周。在室温下储存可能会导致不可靠的结果。
A test procedure for PSTVd is described based on immobilisation of plant sap on filter paper, by dotting or tissue printing followed by RT-PCR. Tests were carried out using primarily and secondarily infected potato plants, primarily infected in vitro plants, and potato tubers. Print PCR was shown to be suitable for testing large samples of potato plants whereas dot PCR is recommended for in vitro plantlets and tuber tissue. Bulking one infected plant to 4 or 9 healthy plants gave reliable results with secondarily infected potato plants, but sometimes the test failed to detect PSTVd in primarily infected in vitro plants. Dotted and printed paper squares could be stored at 4°C for at least 2 weeks in Triton X-100 solution or under dry conditions. Storing at room temperature can lead to unreliable results.