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

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

XIE Xiaofang1,2, TIAN Xianfeng1, JIANG Changjun1,2, LI Yeyun1,*   

  1. 1. State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, China;
    2. Collaborative Innovation Center of Agroforestry Industry in Dabieshan Area, Hefei 230036, China
  • Received:2015-06-26 Revised:2015-10-10 Online:2015-12-15 Published:2019-08-26

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, reference gene, qRT-PCR

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