草业学报2017，Vol.26Issue(3)：75-81,7.DOI:10.11686/cyxb2016297
草地早熟禾荧光定量PCR分析中内参基因的筛选
Screening of reference genes for real-time fluorescence quantitative PCR in Kentucky bluegrass
摘要
Abstract
To select the most stable reference genes of Kentucky bluegrass (Poa pratensis) for real-time quantitative PCR,we assessed the mRNA expression in different tissues of six traditional candidate reference genes at various leaf development stages,under abiotic stress,or following various hormone applications.Genes assessed included ACT,GAPDH,UBQ,EF-1α,18s rRNA and β-tubulin.The expression stabilities of these six reference genes were evaluated by geNorm,NormFinder and BestKeeper.It was found that mRNA expression of the candidate reference genes differed significantly between the tissue types,developmental stages,hormone applications or under abiotic stress.In Kentucky bluegrass leaves,GAPDH is recommended as the reference gene for analyzing the mRNA expression in different tissues;EF-1α was the most stable gene under abiotic stress;β-tubulin was most consistent with different hormones;ACT was stably expressed in the different leaf development stages.In conclusion,use of the most appropriate reference genes under different conditions would enhance the accuracy analysis of gene expression in the Kentucky bluegrass.These findings may be applicable when selecting reference genes for analysis of mRNA expression in other members of the Poa genus.关键词
草地早熟禾/实时荧光定量PCR/内参基因/稳定性评价Key words
Kentucky bluegrass/quantitative real time PCR/reference genes/expression stability evaluation引用本文复制引用
张兰,檀鹏辉,滕珂,闫蒙举,何春燕,甘露,尹淑霞..草地早熟禾荧光定量PCR分析中内参基因的筛选[J].草业学报,2017,26(3):75-81,7.基金项目
国家自然基金项目(31302016)和深圳市科技计划项目(JSGG20160229155434792)资助. （31302016）
