黄梁木实时荧光定量PCR分析中内参基因的选择
收稿日期: 2018-01-03
网络出版日期: 2018-04-26
基金资助
国家自然科学基金(No.31600525)和广东省科技计划(No.2017B020201008)
Selection and Validation of Reference Genes for Quantitative RT-PCR Analysis in Neolamarckia cadamba
Received date: 2018-01-03
Online published: 2018-04-26
为筛选黄梁木(Neolamarckia cadamba)实时定量PCR最佳内参基因, 该研究以黄梁木的根、芽、叶、花、果、皮及形成层为材料, 利用RT-qPCR技术对ACT、CAC、CYP和EF1α等21个管家基因家族43个候选内参基因进行表达量分析, 并利用geNorm、NormFinder和BestKeeper软件进行内参基因稳定性分析。geNorm的分析结果显示, UPL基因的稳定性最高(M=0.443), UBQ基因的稳定性最低(M=2.859); NormFinder的分析结果显示, UPL基因的稳定性最高(E=0.223), UBQ基因的稳定性最低(M=4.759); BestKeeper分析显示, UPL基因的标准偏差(SD=0.513)最低。研究结果表明, UPL基因作为内参基因稳定性最高, UBQ基因的稳定性最低。因此可以选择UPL基因作为黄梁木不同组织中RT-qPCR定量分析的内参基因。
张登, 李景剑, 张梦洁, 包钰韬, 杨霄, 徐武云, 欧阳昆唏, 陈晓阳 . 黄梁木实时荧光定量PCR分析中内参基因的选择[J]. 植物学报, 2018 , 53(6) : 829 -839 . DOI: 10.11983/CBB18003
In this study, root, shoot, leaf, flower, fruit, peel and cambium tissue of Neolamarckia cadamba was sampled to analyze the expression of 43 candidate reference genes in 21 housekeeping gene families, such as ACT, CAC, CYP, and EF1α, by RT-qPCR. The software geNorm, NormFinder and BestKeeper were used to analyze expression stability of these candidate reference genes in the seven different tissues. geNorm analysis revealed that the stability of the UPL gene was the highest (M=0.443) and the stability of UBQ was the lowest (M=2.859). NormFinder analysis revealed that the stability of UPL was the highest (E=0.223), UBQ was the lowest (M=4.759). BestKeeper analysis revealed that the standard deviation of UPL (SD=0.513) was the lowest. These findings suggest that UPL has the highest stability, and UBQ has the lowest stability. UPL gene could be selected as an internal reference gene for analysis of gene expression among different tissues of N. cadamba.
Key words： Neolamarckia cadamba; real-time quantitative PCR; reference gene
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