授粉后红球姜雌性生殖器官qRT-PCR的内参基因筛选

发布时间：2019-06-12 04:49

【摘要】：根据授粉后不同时间点的红球姜转录组数据库以及相关文献报道的传统内参基因,筛选出10个表达相对稳定的基因Actin-2(ACT2)、Actin-7(ACT7)、Beta tubulin-1(TUB1)、Beta tubulin-5(TUB5)、Alpha tubulin-3(TUA3)、Ubiquitin(UBQ)、Glyceraldehyde-3-phosphate dehydrogenase(GAPDH)、Elongation factor 1-alpha(EF-1α)、Cyclophilin(CYP)、Histone(H2A)作为候选内参基因,采用qRT-PCR技术,结合GeNorm、NormFinder和BestKeeper软件对候选内参基因的表达稳定性进行分析.结果表明,在红球姜雌性生殖器官授粉后的发育过程中,GAPDH和UBQ的表达稳定性最好,均适合作为内参基因,同时使用2种作为内参基因能使实时荧光定量PCR标准化分析结果更精确.因此,最终选择GAPDH和UBQ作为实时荧光定量PCR标准化分析红球姜雌性生殖器官相关基因表达的内参基因.该研究将为探究红球姜败育的分子机理奠定基础,也为近源姜属植物内参基因的筛选提供线索.
[Abstract]:According to the database of red ginger transcription group and the traditional internal reference genes reported in the literature, 10 relatively stable genes Actin-2 (ACT2) and Actin-7 (ACT7), Beta tubulin-1 (TUB5), Alpha tubulin-3 (TUA3), Ubiquitin (UBQ), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Elongation factor 1-alpha (EF-1 伪), Cyclophilin (CYP), Histone (H2A) were selected as candidate internal reference genes. QRT-PCR technique and GeNorm,NormFinder and BestKeeper software were used to analyze the expression stability of candidate internal reference genes. The results showed that the expression stability of GAPDH and UBQ was the best during the development of female reproductive organs of Ginger, and both of them were suitable for internal reference genes. At the same time, the results of real-time fluorescence quantitative PCR standardization analysis could be more accurate by using two kinds of internal reference genes as internal reference genes. Therefore, GAPDH and UBQ were selected as internal reference genes for real-time fluorescence quantitative PCR standardization to analyze the expression of genes related to female reproductive organs of Ginger. This study will lay a foundation for exploring the molecular mechanism of abortion of Ginger, and also provide clues for the screening of internal reference genes in the genus Ginger.
【作者单位】：华南师范大学生命科学学院广东省植物发育与生物工程重点实验室;
【基金】：国家自然科学基金委员会-广东省人民政府联合基金(U1301213)
【分类号】：S682.19;Q943.2

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