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Absolute quantification of microRNAs in green tea ( Camellia sinensis ) by stem‐loop quantitative real‐time PCR
Author(s) -
Hou YingHui,
Jeyaraj Anburaj,
Zhang Xiao,
Wei ChaoLing
Publication year - 2017
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.8137
Subject(s) - camellia sinensis , microrna , green tea , real time polymerase chain reaction , biology , endogeny , computational biology , food science , botany , genetics , gene , biochemistry
BACKGROUND There are some studies to show that food‐derived plant microRNAs ( miRNAs ) may be detected in mammals. The research evidence has provoked a considerable debate whether plant‐derived miRNAs exert the same regulatory functions as endogenous animal miRNAs . To test the hypothesis, methods of highly sensitive absolute quantification miRNAs have been developed. However, absolute miRNA quantification of green tea has not yet been reported. This study is the first to build an absolute quantification method to detect miRNAs level in green tea using stem‐loop quantitative real‐time PCR ( qRT‐PCR ). RESULTS Two miRNAs , csn‐ miR164 (a conserved miRNA ) and csn‐ miRn329 (a tea‐specific miRNA ), were selected as examples for the detection and absolute quantification of miRNAs in green tea samples using stem‐loop qRT‐PCR . The content of csn‐ miR164 was significantly higher in the Yuexi Cuilan ( YX ) samples than in the Shucheng Orchid ( SC ) samples. The content of csn‐ miRn329 was found to be high at the start of processing in leaf tissues in both the withering and soaking experiments, after which it gradually decreased with time. CONCLUSION To the best of our knowledge, this is the first report to absolutely quantify the miRNAs present in green tea. This method will help to further investigate the possibility that tea‐derived miRNAs may play an important role on defending against various diseases in humans. © 2016 Society of Chemical Industry