| 落叶松实时定量PCR内参基因的筛选 |
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| 引用本文： | 吴涛,李万峰,张俊红,韩素英,杨文华,齐力旺.落叶松实时定量PCR内参基因的筛选[J].林业科学研究,2013,26(S1):1-8. |
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| 作者姓名： | 吴涛 李万峰 张俊红 韩素英 杨文华 齐力旺 |
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| 作者单位： | 中国林业科学研究院林业研究所, 北京 100091;国家林业局云南珍稀濒特森林植物保护和繁育重点实验室, 云南省森林植物培育与开发利用重点实验室, 云南 昆明 650201;中国林业科学研究院林业研究所, 北京 100091;中国林业科学研究院林业研究所, 北京 100091;中国林业科学研究院森林生态环境与保护研究所, 北京 100091;中国林业科学研究院林业研究所, 北京 100091;中国林业科学研究院林业研究所, 北京 100091 |
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| 基金项目： | 国家"973"计划项目(2009CB119106);国家自然科学基金(30830086,31200464);国家"863"计划项目(2011AA100203,2013AA102704) 资助 |
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| 摘 要： | 本研究针对日本落叶松(Larix kaempferi (Lamb.) Carr.)体细胞胚胎发育过程中原胚团时期到胚胎成熟时期、种子萌发过程、植株幼年生长阶段和成年生长阶段等生长发育过程中的31份材料,应用实时荧光定量PCR(qPCR)技术,分析了12个持家基因(ACT 4、APm、Chc、Gapc、RPL1、RPL2、EF1、EF2、eIF、E3UL、UBQ和UPL2) 在这31份材料中的表达情况。经geNorm和NormFinder两种分析软件对这12个持家基因进行表达稳定性分析,结果表明:APm在不同器官、体细胞胚胎发育和种子萌发过程中表达均最为稳定;EF1 在不同生长年龄植株的顶梢中表达最为稳定;eIF在不同年龄植株的针叶中表达最为稳定。分析结果表明,针对不同的研究材料和发育阶段应选择适合的持家基因做内参基因使用。进行基因表达细微差异研究时,可根据需要使用2个或2个以上内参组合。
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| 关 键 词： | 落叶松 持家基因 qPCR 基因表达 稳定性 |
| 收稿时间： | 2013/8/22 0:00:00 |
Stability Analysis of Reference Genes for Larch Gene Expression Studies by Quantitative Real-Time PCR |
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| WU Tao,LI Wan-feng,ZHANG Jun-hong,HAN Su-ying,YANG Wen-hua and QI Li-wang.Stability Analysis of Reference Genes for Larch Gene Expression Studies by Quantitative Real-Time PCR[J].Forest Research,2013,26(S1):1-8. |
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| Authors: | WU Tao LI Wan-feng ZHANG Jun-hong HAN Su-ying YANG Wen-hua and QI Li-wang |
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| Institution: | Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China;Yunnan Laboratory for Conservation of the Rare, Endangered and Endemic Forest Plants of State Forestry Administration, Yunnan Key Laboratory for Forest Plant Cultivation and Utilization; Yunnan Academy of Forestry, Kunming 650201, Yunnan, China;Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China;Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China;Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing 100091, China;Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China;Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China |
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| Abstract: | The selection of suitable reference gene is an important prerequisite for successful gene expression analysis by quantitative real-time PCR (qPCR). The authors investigated the expression stability of 12 endogenous house-keeping genes (ACT4, APm, Chc, Gapc, RPL1, RPL2, EF1, EF2, eIF, E3UL, UBQ and UPL2) in 31 samples of Larix kaempferi (Lamb.) Carr. including needles, stems, roots, pollens, zygotic embryos and somatic embryos. The GeNorm and NormFinder algorithms analysis reveals that the APm is the most stable gene among different organs and developmental stages (somatic embryogenesis and seed germination), EF1 and eIF are most stable among stems and needles during plant growth (90 days, 1.5 years, 5 years, 10 years, 25 years and 50 year), respectively. The results would provide optimum internal reference genes for larch gene expression analysis using qPCR. |
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| Keywords: | Larix house-keeping gene qPCR gene expression stability |
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