大针茅干旱胁迫下最佳内参基因组合的筛选及验证

摘要

经 qRT-PCR,采用ΔCt 值分析法、geNorm 和 NormFinder 软件分析法综合比较 Act 2、TUB-α、TUB-β、18S rRNA 、GAPDH 、EF-1a 、RNA POL I I 、APRT 、TLF 9个基因的稳定值,筛选大针茅干旱胁迫下基因表达分析最佳内参基因组合。结果表明,在根中最稳定的内参基因组合为18S rRNA 和 EF-1a ,在叶中最稳定的内参基因组合为18S rRNA 和 TLF 。在不同干旱胁迫下用大针茅根和叶中 P IP 2-1和 P IP 2-2基因表达差异验证最佳内参基因组合的稳定性表明,所筛选的内参基因稳定性较好,可以作为大针茅干旱胁迫基因表达的内参基因。%In order to provide stable reference genes for the analysis of gene expression of Stipa gran-dis under the drought stress,stable values of 9 reference genes of Act 2,TUB-α,TUB-β,18S rRNA ,GAP-D H ,EF-1a ,RNA POL I I ,APRT and TLF were investigated by qRT-PCR with ΔCt analyzing method and geNorm and NormFinder software.The best reference genes combination was selected.The results showed that the most stable reference genes combination was 18S rRNA and EF-1a in the root as well as 18S rRNA and TLF in the leaf.The best reference genes combination of P I P 2-1 and P I P 2-2 in the root and leaf of Stipa grandis was proved to be stable under different drought stress.