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以毛白杨(Populus tomentosa)‘BJHR01’的不同组织器官为材料,运用实时荧光定量PCR(qRT-PCR)技术及内参基因表达稳定性分析软件Best Keeper、Norm Finder和geNorm分析了8个看家基因在毛白杨不同组织器官中的表达稳定性。所选看家基因包括细胞骨架蛋白肌动蛋白基因(actin)、微管蛋白基因(TUB)、18S核糖体RNA基因(18S rRNA)、翻译延伸因子(EF-1 beta)、翻译起始因子(eIF 5AV)、组蛋白编码基因(H3)、核糖体蛋白基因(RP)和泛素蛋白基因(UBQ),结果发现eIF 5AV、actin和UBQ三个内参基因在毛白杨不同组织器官中表达水平稳定,适合在定量PCR分析中作为内参基因;TUB的表达稳定性最差,不适宜作为毛白杨不同组织器官基因表达研究的内参基因。在毛白杨不同组织器官基因表达水平研究中,可选用eIF 5AV、actin和UBQ三个内参基因的几何平均数作为参照标准,实验数据的分析将更为准确。
In this study, we used the different tissues and organs of Populus tomentosa ’BJHR01’ as materials and analyzed the expression of 8 housekeeping genes using real-time quantitative PCR (qRT-PCR) and internal reference gene expression stability analysis software Best Keeper, Norm Finder and geNorm Expression stability in different tissues and organs of Populus tomentosa. Selected housekeeping genes include the actin, TUB, 18S rRNA, EF-1 beta, translation initiation factor ( eIF 5AV, H3, Rb and UBQ. The results showed that the expression of eIF 5AV, actin and UBQ were stable in different tissues and organs of Populus tomentosa , Suitable for quantitative PCR analysis as a reference gene; TUB expression of the worst stability, inappropriate Populus tomentosa different tissues and organs gene expression study of the internal reference gene. In the study of the gene expression of different tissues and organs of Populus tomentosa, the geometric mean of three internal reference genes eIF 5AV, actin and UBQ can be used as the reference standard, the analysis of experimental data will be more accurate.