文冠果种子发育基因表达分析内参基因的选择

梁重钧; 王利兵; 张鑫; 姜国维; 宁楚龙; 李麟坤; 张薇

doi:10.12403/j.1001-1498.20230291

文冠果种子发育基因表达分析内参基因的选择

Selection of Reference Genes for Gene Expression Analysis in Seed Development of Xanthoceras sorbifolium Bunge

摘要

摘要:
目的验证ACT基因是否适用于文冠果种子发育的相关基因表达分析，并筛选出在文冠果基因表达分析中最稳定的内参基因。
方法本研究采用了4种数学算法（ΔC_T、BestKeeper、NormFinder、geNorm）评估12个候选内参基因的表达稳定性，并利用在线工具RefFinder综合评价。
结果在文冠果不同发育时期种子内参基因综合评分排名为PP2A＞Tip41> EF-1α＞18r＞UCE＞β-TUB＞UBQ＞α-TUB＞CYP＞SAM＞ACT＞GADPH；在文冠果不同组织内参基因综合评分排名为PP2A＞Tip41>CYP＞18r＞UBQ＞SAM＞EF-1α＞UCE＞GADPH＞α-TUB＞ACT＞β-TUB。
结论 PP2A和Tip41基因是文冠果基因表达分析最稳定的内参基因。

Abstract:
Objective : To assess the suitability of the ACT gene for gene expression analysis in seed development of Xanthoceras sorbifolium, and to identify the most stable reference genes for gene expression analysis in X. sorbifolium.
Method : In this study, four mathematical algorithms (ΔC_T, BestKeeper, NormFinder, geNorm) were carried out for evaluating the expression stability of 12 candidate reference genes based on the web-based tool RefFinder.
Result : The RefFinder result showed that the gene PP2A performed the best, followed byTip41, EF-1α,18r, UCE, β-TUB, UBQ, α-TUB, CYP, SAM, ACT, and GADPH at the different seed development stages in X. sorbifolium , and the gene also had the highest score, followed by Tip41, CYP, 18r, UBQ, SAM, EF-1α, UCE, GADPH, α-TUB, ACT, and β-TUB at various tissues.
Conclusion : PP2A and Tip41 are the two most stable reference genes for gene expression analysis in X. sorbifolium.

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