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核桃内参基因实时荧光定量PCR表达稳定性评价 被引量:18

Stability evaluation of reference genes for quantitative real-time PCR analysis in walnut(Juglans spp.)
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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.
作者 李雪 潘学军 张文娥 张睿 陈静 LI Xue PAN Xue-Jun ZHANG Wen-E ZHANG Rui CHEN Jing Guizhou(Engineering Research Center for Fruit Crops, Guizhou University, Guiyang 550025, Chin)
出处 《植物生理学报》 CAS CSCD 北大核心 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)~~
关键词 核桃 实时荧光定量PCR 内参基因 walnut qRT-PCR reference gene
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