| 茶树实时荧光定量PCR分析中内参基因的选择 |
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| 引用本文： | 孙美莲,王云生,杨冬青,韦朝领,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]. 植物学通报, 2010, 45(5): 579-587. DOI: 10.3969/j.issn.1674-3466.2010.05.007 |
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| 作者姓名： | 孙美莲 王云生 杨冬青 韦朝领 高丽萍 夏涛 单育 骆洋 |
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| 作者单位： | [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 |
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| 基金项目： | 973计划前期项目，国家自然科学基金，安徽省自然科学基金 |
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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-tubulin4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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| 关 键 词： | 茶树 实时荧光定量PCR 内参基因 |
Reference Genes for Real-time Fluorescence Quantitative PCR in Camellia sinensis |
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| Meilian Sun,Yunsheng Wang,Dongqing Yang,Chaoling Wei,Liping Gao,Tao Xia,Yu Shan,Yang Luo. Reference Genes for Real-time Fluorescence Quantitative PCR in Camellia sinensis[J]. Chinese Bulletin of Botany, 2010, 45(5): 579-587. DOI: 10.3969/j.issn.1674-3466.2010.05.007 |
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| Authors: | Meilian Sun Yunsheng Wang Dongqing Yang Chaoling Wei Liping Gao Tao Xia Yu Shan Yang Luo |
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| Affiliation: | 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 |
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| 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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| Keywords: | Camellia sinensis real-time fluorescence quantitative PCR reference genes |
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