植物学报 ›› 2010, Vol. 45 ›› Issue (05): 579-587.DOI: 10.3969/j.issn.1674-3466.2010.05.007

• 技术方法 • 上一篇    下一篇

茶树实时荧光定量PCR分析中内参基因的选择

孙美莲1, 王云生2, 杨冬青1, 韦朝领1, 高丽萍2*, 夏涛1*, 单育1, 骆洋2   

  1. 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 出版日期:2010-09-01 发布日期:2010-09-20
  • 通讯作者: 高丽萍;夏涛

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

Meilian Sun1, Yunsheng Wang2, Dongqing Yang1, Chaoling Wei1, Liping Gao2*, Tao Xia1*, Yu Shan1, Yang Luo2   

  1. 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:2009-12-08 Revised:2010-03-30 Online:2010-09-01 Published:2010-09-20
  • Contact: Liping Gao, Tao Xia

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、GAPDHβ-actinα-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。

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.