菊芋实时定量PCR分析中内参基因的选择  

Selection of Reference Genes for Quantitative RT-PCR Analysis of Helianthus tuberosus

作　　者：宋向阳[1] 杨世鹏[1] 钟启文[1,2] 王丽慧[1] 赵孟良[1] 李莉[1,2] 孙雪梅[1,2] 

机构地区：[1]青海大学农林科学院青海省蔬菜遗传与生理重点实验室,西宁810016 [2]青海大学三江源生态和高原农牧业国家重点实验室,西宁810016

出　　处：《分子植物育种》2018年第4期1190-1196,共7页Molecular Plant Breeding

基　　金：国家自然科学基金资助项目(31460523;31660569;31660588);青海省科技厅项目(N2016-ZJ-751;2015-HZ-805)共同资助

摘　　要：实时荧光定量PCR具有灵敏度、特异性和重复性好等优点,是植物中常用于基因表达和转录分析的重要手段。在分子生物学的研究中,选择表达相对稳定的内参基因是分析实时荧光定量实验数据的前提。本研究以菊芋(Helianthus tuberosus)开花时期的幼根、嫩茎、叶片、块茎和花瓣为材料,运用荧光定量PCR方法分析了18S rRNA、EF1α、Actin、β-actin、GAPDH、25S rRNA和UBQ5 7个内参基因的表达量,通过Ge Norm、Norm Finder和Best Kkeeper软件的综合分析,发现25S r RNA的稳定性最好,可用于菊芋基因表达的内参基因。而GAPDH在不同样品中稳定性最差,不适合作为内参基因。本研究为菊芋实时荧光定量PCR中内参基因的选择提供了参考。Real-time quantitative PCR has the advantages of sensitivity, specificity and reproducibility, which is an important method commonly used in gene expression and transcription analysis. In the study of molecular biology, the choice of relatively stable reference gene is a prerequisite for the analysis of real-time quantitative data. In this study, the expression content of seven reference genes including 18S rRNA, EF1α, Actin, 13-actin, GAPDH, 25S rRNA and UBQ5 were analyzed by quantitative PCR using the roots, stems, leaves, tubers and petals in flowering period ofHelianthus tuberosus. The comprehensive analysis ofGeNorm, NormFinder and BestKeeper found that the stability of 25S rRNA was the best, which could be used as the internal reference gene for the gene expression of Helianthus tuberosus. While GAPDH had the worst stability in the different samples, so it was not suitable as the reference gene. This study could provide a reference for the selection of internal reference genes for real-time fluorescence quantitative PCR ofHelianthus tuberosus.

关 键 词：菊芋 实时荧光定量PCR 内参基因 

分 类 号：S632.9[农业科学—蔬菜学]