文冠果实时荧光定量PCR内参基因的筛选

为筛选适用于文冠果(Xanthoceras sorbifolium Bunge)性别分化过程研究和不同器官的内参基因,本研究以文冠果性别分化关键期的顶芽和侧芽、根、茎、叶、种仁、雌能花和雄能花为材料,通过实时荧光定量PCR(qRT-PCR)鉴定7个候选内参基因(Actin,UBQ,EF-1α,TUB,eIF-4α,18S rRNA和GAPDH)在各样品中的表达量,并评价7个基因的表达稳定性。结果表明,7个候选内参基因在不同样品中的表达量存在明显差异,各基因在雌能花中的表达均显著高于其他材料,在雄能花中的表达处于较低水平;适用于文冠果性别分化过程的最优内参基因为UBQ和eIF-4α,适用于不同器官的最优内参基因为EF-1α和GAPDH。研究结果将对今后进行文冠果性别分化过程研究和不同器官中的基因表达分析提供参考。

Selection of Reference Genes for Quantitative Real-Time PCR in Xanthoceras sorbifolium Bunge

In order to select the reference genes suitable for sex differentiation process and different organs of Xanthoceras sorbifolium Bunge,the apical and lateral buds at the critical period of sex differentiation,roots,stems,leaves,seed kernels,female flowers,and male flowers of X.sorbifolium were taken as materials in this study.Quantitative real-time PCR(qRT-PCR)was used to identify the expression levels of seven candidate reference genes(Actin,UBQ,EF-1α,TUB,e IF-4α,18S rRNA and GAPDH)in each material,and the stability of these 7 genes was evaluated.The results showed that the expression levels of the 7 candidate reference genes were obviously different in different materials.The expression of each gene in female flowers was significantly higher than that in other materials,and the expression in male flowers was at a lower level;UBQ and eIF-4αwere the optimal reference genes for sex differentiation process,and EF-1αand GAPDH were the best reference genes suitable for different organs.The results of this study would provide a reference for the future research on the gene expression analysis in sex differentiation process and different organs of X.sorbifolium.