《植物生理学报》 2017, 53(9): 1795-1802

核桃内参基因实时荧光定量PCR表达稳定性评价

李雪, 潘学军^*, 张文娥^*, 张睿, 陈静

贵州大学/贵州省果树工程技术研究中心, 贵阳550025

通信作者：潘学军；E-mail: pxjun2050@aliyun.com; agr.wezhang@gzu.edu.cn

摘　要：

利用实时荧光定量PCR技术, 结合geNorm、NormFinder和BestKeeper软件研究了5个常用的植物内参基因GAPDH、β-Actin、18S rRNA、UBQ和EF-1在不同基因型、不同组织、不同发育时期核桃(Juglans spp.)中的表达情况, 以期筛选出在核桃中稳定表达的内参基因。结果显示: GAPDH和EF-1在核桃所有样品中均表达稳定, 由于表达丰度偏低, 所以适合作为分析较低表达丰度目标基因时的内参; 18S rRNA在核桃所有样品中表达稳定性较好, 且18S rRNA表达丰度偏高, 适合作为分析较高表达丰度目标基因时的内参; β-Actin和UBQ在核桃样品中的表达稳定性较差, 不适合作为核桃中基因表达分析时的内参基因。

关键词：核桃; 实时荧光定量PCR; 内参基因

收稿：2017-06-05   修定：2017-08-08

资助：贵州省高层次创新型人才培养项目(黔科合人才[2016]4038号)和贵州大学研究生创新基金(研农2017019)。

Stability evaluation of reference genes for quantitative real-time PCR analysis in walnut (Juglans spp.)

LI Xue, PAN Xue-Jun^*, ZHANG Wen-E^*, ZHANG Rui, CHEN Jing

Guizhou Engineering Research Center for Fruit Crops, Guizhou University, Guiyang 550025, China

Corresponding author: PAN Xue-Jun; E-mail: pxjun2050@aliyun.com; agr.wezhang@gzu.edu.cn

Abstract:

In order to screen out the ideal reference genes for stable expression in walnut (Juglans spp.), GAPDH, β-Actin, 18S rRNA, UBQ and EF-1 were used to evaluate the expression stability in different walnut genotypes, tissues, and developmental stages by using quantitative real-time PCR and softwares of geNorm, Norm-Finder and BestKeeper. The results show that reference genes GAPDH and EF-1 were identified as stable genes in all samples of walnut, followed by 18S rRNA, β-Actin and UBQ. GAPDH and EF-1 were suitable for the analysis of lower target genes due to their low abundance of expression. 18S rRNA abundance of expression is high and suitable for analysis of higher expression target genes. However, the expression stability of β-Actin and UBQ in all samples was poor, which was not suitable as reference genes in walnut.

Key words: walnut; qRT-PCR; reference gene