橡胶草实时荧光定量PCR内参基因评价

Evaluation of reference genes for quantitative real-time PCR expression studies in Taraxacum kok-saghyz Rodin

橡胶草(Taraxacum kok-saghyz)是一种重要的产胶作物。为了筛选出橡胶草稳定可靠的内参基因用于实时荧光定量PCR(qRT-PCR)分析,本研究从已公布的橡胶草转录组数据中选取18S rRNA、EIF4α、tubulin、β-actin、GAPDH和40S ribosomal protein(40SRP)作为候选基因,通过qRT-PCR检测,并利用geNorm、Norm Finder、Best Keeper等软件对这6个候选内参基因在不同橡胶草种质、不同组织以及机械伤害和乙烯利处理后不同时间下的表达稳定性进行分析,结果发现6个候选内参基因的表达稳定性各异,GAPDH和β-actin基因是不同种质及其不同组织以及不同处理中表达最稳定的内参基因。本研究结果为橡胶草基因表达分析提供了稳定可靠的内参基因。

Taraxacum kok-saghyz（TKS） is one of the most important rubber-producing crops. To screen stable and reliable reference genes for quantitative real-time PCR（q RT-PCR） analysis in TKS, 6 housekeeping genes from the public transcriptome data of TKS, including 18 S rRNA, EIF4α, tubulin, β-actin, GAPDH, and 40 S ribosomal protein（40SRP）, were selected as candidate reference genes for analysis in this study. The expression stabilities of the 6 genes were analyzed by geNorm, Norm Finder, and Best Keeper, respectively, in different TKS accessions and tissues, and under mechanical wounding and ethephon treamtents. This result indicates that the expression stability varied greatly between these genes. GAPDH and β-actin had the highest stabilities across different accessions and tissues, and under different treatments. The results in this study provide stable and reliable reference genes for gene expression analysis in TKS.