羊草不同组织实时定量PCR 内参基因的筛选

摘要

为筛选羊草(Leymus chinensis)不同组织实时定量PCR(qRT-PCR)试验体系中的最佳内参基因,以羊草叶、茎、根、穗为研究材料,利用qRT-PCR技术分析TUA、TUB、18S rRNA、EF-1α、APRT、CYP、Actin和CBP20共8个常用候选管家基因的表达情况,并使用geNorm和NormFinder软件进行综合评价,以期筛选出羊草不同组织中最稳定的内参基因.结果表明,8个候选内参基因在羊草不同组织表达稳定性不同,其中,叶片中稳定性较高的内参基因是Actin;茎中稳定性较高的内参基因是EF-1a;根中稳定性较高的内参基因是APRT和18S rRNA;穗中稳定性较高的内参基因是TUB.本研究结果将为开展羊草功能基因的表达分析提供重要参考.%To explore the optimal reference genes in different tissues of Leymus chinensis for a quantitative real-time PCR experiment, eight general housekeeping genes including TUA, TUB, 18S rRNA, EF-1α, APRT, CYP, Actin and CBP20 were selected as candidate reference genes.Using roots, stems, leaves, and spikes of L.chinensis, we analyzed the 8 candidate reference gene expressions using qRT-PCR and evaluated their expression stability using geNorm and NormFinder software.The results showed that stability differed significantly among the eight candidate reference genes in different tissues.The most stable reference gene was Actin in leaves, EF-1a in stems, APRT and 18S rRNA in root and TUB in spikes.The present study has provided an important reference for analysis the expression of functional genes in L.chinensis.