茶树低温胁迫下microRNA实时定量PCR内参基因的筛选

doi:10.13305/j.cnki.jts.2015.06.013

茶叶科学 ›› 2015, Vol. 35 ›› Issue (6): 596-604.doi: 10.13305/j.cnki.jts.2015.06.013

茶树低温胁迫下microRNA实时定量PCR内参基因的筛选

谢小芳^1,2, 添先凤¹, 江昌俊^1,2, 李叶云^1,*

1. 安徽农业大学茶树生物学与资源利用国家重点实验室,安徽合肥 230036;
2. 安徽农业大学大别山区农林特色产业协同创新中心,安徽合肥 230036

收稿日期:2015-06-26 修回日期:2015-10-10 出版日期:2015-12-15 发布日期:2019-08-26
通讯作者: *lyy@ahau.edu.cn
作者简介:谢小芳,女,硕士,主要从事茶树种质资源与育种技术方面的研究。
基金资助:
国家自然科学基金（31270729）

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,*

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

摘要/Abstract

摘要： MicroRNA（miRNA）在基因的转录后调控上起到重要作用,对其表达水平的定量研究已成为一种常见实验,选择合适的内参基因是确保实时荧光定量PCR（qRT-PCR）准确性的先决条件。本研究以茶树为材料,挑选了9个候选内参基因,用qRT-PCR技术检测它们在不同样本中的表达量,采用BestKeeper和geNorm软件进行表达稳定性综合分析。结果表明,一种茶树新miRNA（PC-3p-222）在不同低温处理以及不同茶树组织中表达最为稳定,Ct平均值为22~23,丰度适中,可以作为茶树低温胁迫下miRNA qRT-PCR的内参基因,为miRNA定量研究工作提供参考。

关键词: 茶树, miRNA, 内参基因, qRT-PCR

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

中图分类号:

S571.1
Q52

谢小芳, 添先凤, 江昌俊, 李叶云. 茶树低温胁迫下microRNA实时定量PCR内参基因的筛选[J]. 茶叶科学, 2015, 35(6): 596-604. doi: 10.13305/j.cnki.jts.2015.06.013.

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. doi: 10.13305/j.cnki.jts.2015.06.013.

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茶树低温胁迫下microRNA实时定量PCR内参基因的筛选

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

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