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Rapid Detection of Tomato chlorosis virus from Infected Plant and Whitefly by One‐step Reverse Transcription Loop‐mediated Isothermal Amplification
Author(s) -
Karwitha Miriam,
Feng ZhiKe,
Shen Yan,
Essendi Walter,
Zhang WenNa,
Li JinYu,
Tao XiaoRong
Publication year - 2016
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/jph.12453
Subject(s) - reverse transcription loop mediated isothermal amplification , loop mediated isothermal amplification , biology , chlorosis , whitefly , agarose , virology , plant virus , rna , sybr green i , reverse transcriptase , virus , population , microbiology and biotechnology , horticulture , real time polymerase chain reaction , genetics , gene , dna , demography , sociology
Tomato chlorosis virus (To CV ) is the causal agent of an emerging virus disease of tomatoes which causes significant economic losses worldwide. The rapid spread of To CV is associated with the increasing whitefly population in many countries. In this study, a highly efficient and practical one‐step reverse transcription loop‐mediated isothermal amplification ( RT ‐ LAMP ) was developed for the rapid detection of To CV in infected plant or whitefly. This RT ‐ LAMP assay can detect To CV either in total RNA or crude RNA extracted from infected plants using a water bath within 1 h. The presence of To CV in RT ‐ LAMP products could be evaluated as ladder‐like bands in an agarose gel or visualized in‐tube with inclusion of a SYBR Green I dye under UV or daylight. RT ‐ LAMP was 100–1000 times more sensitive compared to conventional RT ‐ PCR for the detection of To CV . RT ‐ LAMP amplification was specific to To CV , and no false reactions were detected when other viruses were tested. The developed RT ‐ LAMP assay was also able to detect To CV from purified RNA extracted from its vector whitefly. This is the first report of the application of the RT ‐ LAMP to detect To CV . The RT ‐ LAMP assay developed in this study provides a rapid, sensitive and practical approach to facilitate the surveillance and management of To CV .