刺槐实时定量PCR分析中内参基因的选择

doi:10.11707/j.1001-7488.20140923

林业科学 ›› 2014, Vol. 50 ›› Issue (9): 167-172.doi: 10.11707/j.1001-7488.20140923

刺槐实时定量PCR分析中内参基因的选择

王金星¹, 张利军², 廖资亿³, 张运根⁴, 邱乾栋¹, 孙鹏^{1 5}, 孙宇涵¹, 胡瑞阳¹, 卢楠¹, 李云¹

1. 林木育种国家工程实验室林木、花卉遗传育种教育部重点实验室北京林业大学生物科学与技术学院北京 100083;
2. 北京风沙源育苗中心北京 102115;
3. 福建省三明市国有林场管理处三明 365000;
4. 福建省宁化国有林场宁化 365413;
5. 中国林业科学研究院经济林研究开发中心郑州 450003

收稿日期:2013-12-04 修回日期:2014-03-28 出版日期:2014-09-25 发布日期:2014-09-30
基金资助:
国家自然科学基金项目“刺槐杂交结实率低的机理研究”（31170629）；国家科技支撑项目“抗逆生态树种刺槐新品种选育技术研究”（2012BAD01B0601）；北京市科委项目“首都平原百万亩造林科技支撑工程”（Z121100008512002）。

The Selection of Reference Genes for Real-Time Quantitative PCR Normalization in Black Locust(Robinia pseudoacacia)

Wang Jinxing¹, Zhang Lijun², Liao Ziyi³, Zhang Yungen⁴, Qiu Qiandong¹, Sun Peng^{1 5}, Sun Yuhan¹, Hu Ruiyang¹, Lu Nan¹, Li Yun¹

1. National Engineering Laboratory for Tree Breeding Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education College of Biological Sciences and Technology, Beijing Forestry University Beijing 100083;
2. Beijing Sandstorm Source Nursery Center Beijing 102115;
3. Administrative Department of State-Owned Forest Farm of Sanming City in Fujian Province Sanming 365000;
4. The State-Owned Forest Farm of Ninghua County in Fujian Province Ninghua 365413;
5. Non-Timber Forestry Research and Development Center, Chinese Academy of Forestry Zhengzhou 450003

Received:2013-12-04 Revised:2014-03-28 Online:2014-09-25 Published:2014-09-30
Contact: 李云

摘要/Abstract

关键词: 刺槐, 内参基因, 实时荧光定量PCR, GeNorm, NormFinder

Abstract:

The real-time PCR is an important method for analyzing gene expression levels. Selection of reference genes suitable for calibration of the expression of objective gene according to specific experimental materials and conditions is a prerequisite for obtaining reliable RT-PCR results. In this study, we used different individuals and different organs of black locust(Robinia pseudoacacia)to test the stability of six internal genes such as Actin gene (ACT), 18S ribosomal RNA ( 18S rRNA), adenosylmethionine decarboxylase gene (SAMDC), helicase gene (Helicase), elongation factor gene ( EF1 -α), glyceraldehyde-3-phosphate-dehydrogenase gene (GAPDH) in the real-time PCR. After being analyzed with Genorm and NormFinder, the results showed that the ACT and 18S rRNA gene were the most stable genes by using different plant individuals as experimental material, while the ACT and GAPDH were the most stable genes with using different tissues and organs. Since some controversies existed with the 18S rRNA as reference gene, the both ACT and GAPDH used for reference gene in various test conditions obtained more accurate results. This study has important implications in obtaining a more accurate result of quantitative expression of black locust.

Key words: Robinia pseudoacacia, reference gene, real-time quantitative PCR, GeNorm, NormFinder

中图分类号:

S718.46

王金星, 张利军, 廖资亿, 张运根, 邱乾栋, 孙鹏, 孙宇涵, 胡瑞阳, 卢楠, 李云. 刺槐实时定量PCR分析中内参基因的选择[J]. 林业科学, 2014, 50(9): 167-172.

Wang Jinxing, Zhang Lijun, Liao Ziyi, Zhang Yungen, Qiu Qiandong, Sun Peng, Sun Yuhan, Hu Ruiyang, Lu Nan, Li Yun. The Selection of Reference Genes for Real-Time Quantitative PCR Normalization in Black Locust(Robinia pseudoacacia)[J]. Scientia Silvae Sinicae, 2014, 50(9): 167-172.

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刺槐实时定量PCR分析中内参基因的选择

The Selection of Reference Genes for Real-Time Quantitative PCR Normalization in Black Locust(Robinia pseudoacacia)

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