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目的筛选合适的内参基因用于青天葵实时荧光定量PCR分析的校正。方法以毛唇芋兰3个组织(叶片、叶柄和球茎)为材料,利用实时荧光定量PCR技术探讨18S rRNA、actin、ubiquitin、EF-1α和β-tubulin 5个常用内参基因在毛唇芋兰不同组织中表达差异。利用GeNorm和NormFinder软件比较各内参基因的Ct值,以分析他们在青天葵3个组织的表达稳定性。结果 5个内参基因的表达稳定性各异,GeNorm软件分析结果表明,稳定性β-tubulin=EF-1α>ubiquitin>actin>18S rRNA;NormFinder软件结果显示β-tubulin稳定性最好,EF-1α次之,18S rRNA则最差。两个不同软件分析结果一致。结论β-tubulin可作为毛唇芋兰不同组织中基因表达差异分析的校正内参基因。
Objective To screen a suitable internal reference gene for the correction of real-time quantitative PCR analysis of Artemisia glauca. Methods Three tissues (leaves, petioles and bulbs) of Polygonatum konjac were used as materials. Five commonly used internal reference genes of 18S rRNA, actin, ubiquitin, EF-1α and β-tubulin were detected by real-time fluorescence quantitative PCR Differences in expression in different tissues. The Ct values of each internal reference gene were compared using GeNorm and NormFinder software to analyze the stability of their expression in three tissues of Araceae. Results The results of GeNorm software showed that the stability of β-tubulin was the best. The results of NormFinder software showed that the stability of β-tubulin was the highest, EF-1α Second, 18S rRNA is the worst. Two different software analysis of the same. Conclusion β-tubulin can be used as a corrective reference gene for differential gene expression analysis in different tissues of P. koraiensis.