红掌佛焰苞基因qRT-PCR分析中内参基因的筛选

Screening of Reference Genes in Anthurium andraeanum Spathes for qRT-PCR Analysis

摘要:

为筛选红掌(Anthurium andraeanum Linden)中稳定表达、可用于佛焰苞中实时荧光定量PCR 分析(qRT-PCR)的内参基因,对5 个组成型表达基因EF1-a、UBQ7、ACTB、GADPH、His3 进行表达稳定性分析,并利用所筛选的内参基因研究红掌的二氢黄酮醇还原酶基因(dfr)的表达水平。结果表明,5 种内参基因在不同品种间的表达稳定性不同。据内参基因标准化因子的配对差异分析(V_n/n+1),判定内参基因的最适数目为2,ACTB 和UBQ7 在红掌不同品种及佛焰苞发育不同阶段中表达均稳定,是理想的内参基因。dfr 在不同品种的佛焰苞及佛焰苞发育过程中均有表达,且dfr 表达水平的变化趋势一致,因此,所选内参基因是合适的。

关键词:红掌;佛焰苞;实时荧光定量PCR;内参基因

Abstract:

In order to screening stable reference genes that can be used to real-time quantitative PCR (qRT-PCR) analysis in Anthurium andraeanum spathes, five constitutively expressed genes, including EF1-a, UBQ7, ACTB, GADPH and His3, were analyzed expression stability by using qRT-PCR, and the expression of dihydroflavonol 4-reductase gene (dfr) was analyzed by using screened reference gene. The results showed that the expression stabilities of five reference genes were different among different cultivars. On the basis of the normalization factor V_n/n+1, the optimum number of reference gene was two. The expression of ACTB and UBQ7 were the most stable in dfr ferent cultivars and different development stages of A. andraeanum spathes, so that they were ideal reference genes. When ACTB and UBQ7 were used as reference genes, the gene dfr expressed in different cultivars and different development stages of A. andraeanum spathes, and the change trend of dfr expression were consistent. Thus, both ACTB and UBQ7 were suitable as reference genes for A. andraeanum spathes.

Key words:Anthunium andraenum;Spathe;qRT-PCR;Reference gene