A rapid sap-direct reverse transcription-polymerase chain reaction method for detection of dendrobium viroid in Dendrobium plants
文献类型： 外文期刊
作者： Li, S. 1 ; Yang, Y. 3 ; Xing, F. 1 ; Che, H. Y. 3 ; Cao, X. R. 3 ; Zhang, Z. X. 1 ; Khoo, Y. W. 1 ; Zhou, C. Y. 2 ; Li, S. F 1 ;
作者机构： 1.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing, Peoples R China
2.Southwest Univ, Chinese Acad Agr Sci, Citrus Res Inst, Chongqing, Peoples R China
3.Chinese Acad Trop Agr Sci, Environm & Plant Protect Inst, Haikou 571101, Hainan, Peoples R China
关键词： Dendrobium; DVd; pipette; reverse transcription‐ polymerase chain reaction; sap; viroid
期刊名称：LETTERS IN APPLIED MICROBIOLOGY （ 影响因子：2.173； 五年影响因子：2.07 ）
ISSN： 0266-8254
年卷期：
页码：
收录情况： SCI
摘要： Dendrobium viroid (DVd) was first reported in China in 2020, and it is the only viroid known to infect Orchidaceae family plants. In this study, we developed a simple reverse transcription-polymerase chain reaction (RT-PCR) method for the rapid detection of DVd in Dendrobium plants. When extracting the sap template from the leaves, they are first clamped between two layers of plastic film, and the sap is pressed out and collected with a pipette. Using this sap, DVd was detected by dot-blot and RT-PCR methods and, the expected amplicons were confirmed by sequencing analysis. The batch analysis of field samples revealed that this method can be used to detect DVd rapidly. The detection method also reduces cross-contamination between different samples and minimizes false positives. Thus, this sap-direct RT-PCR method allows effective and rapid DVd detection in the study of Orchidaceae plants.
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