茶树实时荧光定量PCR分析中内参基因的选择

孙美莲; 王云生; 韦朝领; 杨冬青; 高丽萍; 夏涛; 单育; 骆洋

doi:10.3969/j.issn.1674-3466.2010.05.007

植物学报 >

2010 , Vol. 45 >Issue 05: 579 - 587

DOI: https://doi.org/10.3969/j.issn.1674-3466.2010.05.007

技术方法

茶树实时荧光定量PCR分析中内参基因的选择

孙美莲 ,
王云生 ,
韦朝领 ,
杨冬青 ,
高丽萍 ,
夏涛 ,
单育 ,
骆洋

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¹安徽农业大学农业部茶叶生物化学与生物技术重点实验室, 合肥 230036
²安徽农业大学生命科学学院, 合肥 230036

收稿日期: 2009-12-08

修回日期: 2010-03-30

网络出版日期: 2010-09-20

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Reference Genes for Real-time Fluorescence Quantitative PCR in Camellia sinensis

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¹Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036,China

²School of Biology Science, Anhui Agricultural University, Hefei 230036, China

Received date: 2009-12-08

Revised date: 2010-03-30

Online published: 2010-09-20

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摘要

选择合适的内参基因是提高实时荧光定量PCR分析(qRT-PCR)准确性的先决条件。该文以茶树(Camellia sinensis) 芽、叶、幼根、嫩茎、花瓣、种子和愈伤组织为材料, 应用实时荧光定量PCR技术, 分析了18S rRNA、GAPDH、β-actin和α-tubulin 4个常用内参基因在茶树不同器官组织中的表达情况。经GeNorm和NormFinder软件分析发现, 当利用荧光定量PCR分析比较茶树不同器官组织中的基因表达差异时, 可选择β-actin作为校正内参基因; 而比较不同成熟度的叶片和愈伤组织时, 可以选择GAPDH作为校正内参基因。

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孙美莲 , 王云生 , 韦朝领 , 杨冬青 , 高丽萍 , 夏涛 , 单育 , 骆洋 . 茶树实时荧光定量PCR分析中内参基因的选择[J]. 植物学报, 2010 , 45(05) : 579 -587 . DOI: 10.3969/j.issn.1674-3466.2010.05.007

Abstract

The selection of a suitable reference gene is an important prerequisite for successful gene expression analysis by real-time fluorescence quantitative PCR (qPCR). We investigated the expression stability of 4 endogenous candidate genes (18S rRNA, GAPDH, β-actin and α-tubulin) in qPCR experiments in different organs and tissues, including buds, leaves, young roots, stem, petals, seeds, and callus, of the tea plant Camellia sinensis (L.) O. Kuntze. The analysis with GeNorm and NormFinder algorithms revealed that β-actin could be used as a reference gene for organs and tissues and GAPDH for mature leaves and callus.

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