Screening of microRNA Reference Genes for Real-time Fluorescence Quantitative PCR under Cold Stress in Camellia sinensis

XIE Xiaofang; TIAN Xianfeng; JIANG Changjun; LI Yeyun

doi:10.13305/j.cnki.jts.2015.06.013

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Journal of Tea Science ›› 2015, Vol. 35 ›› Issue (6) : 596-604. DOI: 10.13305/j.cnki.jts.2015.06.013

Screening of microRNA Reference Genes for Real-time Fluorescence Quantitative PCR under Cold Stress in Camellia sinensis

XIE Xiaofang^1,2, TIAN Xianfeng¹, JIANG Changjun^1,2, LI Yeyun^1,*

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Abstract

MicroRNA (miRNA) plays an important role in post transcriptional regulation of gene expression. The research of miRNA quantitative expression level has become more and more popular. Choosing a suitable reference gene is a prerequisite of an accurate real-time fluorescence quantitative PCR(qRT-PCR) assay. The expressions of 9 candidate reference genes by qRT-PCR method in different tea (Camellia sinensis) samples were investigated and their stabilities were analyzed by BestKeeper and geNorm. The results showed that a novel tea miRNA (PC-3p-222) was the most stable reference gene in different tea plant tissues under different cold temperature. The average Ct value was 22-23 which showed moderate expression abundance. Therefore, it can be used as the appropriate reference gene for miRNA qRT-PCR under cold stress, which provides a good choice for the quantitative of miRNA expression.

Key words

Camellia sinensis / miRNA / qRT-PCR / reference gene

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XIE Xiaofang, TIAN Xianfeng, JIANG Changjun, LI Yeyun. Screening of microRNA Reference Genes for Real-time Fluorescence Quantitative PCR under Cold Stress in Camellia sinensis[J]. Journal of Tea Science. 2015, 35(6): 596-604 https://doi.org/10.13305/j.cnki.jts.2015.06.013

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