核桃内参基因实时荧光定量PCR表达稳定性评价  

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

作　　者：李雪[1] 潘学军[1] 张文娥[1] 张睿[1] 陈静[1] LI Xue PAN Xue-Jun ZHANG Wen-E ZHANG Rui CHEN Jing Guizhou(Engineering Research Center for Fruit Crops, Guizhou University, Guiyang 550025, Chin)

机构地区：[1]贵州大学/贵州省果树工程技术研究中心,贵阳550025

出　　处：《植物生理学报》2017年第9期1795-1802,共8页Plant Physiology Journal

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

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。In order to screen out the ideal reference genes for stable expression in walnut（Juglans spp.）, GAPDH, β-Actin, 18 S 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 ge Norm, NormFinder and Best Keeper. The results show that reference genes GAPDH and EF-1 were identified as stable genes in all samples of walnut, followed by 18 S 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. 18 S rRNA abundance of expression is high and suitable for analysis of higher expression target genes. However, the expression stability of β-Actinand UBQ in all samples was poor, which was not suitable as reference genes in walnut.

关 键 词：核桃 实时荧光定量PCR 内参基因 

分 类 号：S664.1[农业科学—果树学]