开发和评估适合枸杞和黑果枸杞杂种衍生 qRT-PCR 归一化的合适参考基因。
Development and evaluation of suitable reference genes for qRT-PCR normalization of hybrids derived from Lycium barbarum and Lycium ruthenicum.
机构信息
Academy of Agriculture and Forestry Sciences, Qinghai University, Xining, 810016, China.
Laboratory for Research and Utilization of Germplasm Resources on the Qinghai-Tibet Plateau, Xining, 810016, China.
出版信息
Mol Biol Rep. 2024 Aug 20;51(1):922. doi: 10.1007/s11033-024-09848-0.
BACKGROUND
A correct and stably expressing reference gene is prerequisite for successful quantitative real-time PCR (qRT-PCR). Investigating gene expression profiling during flower development could enhance our understanding of the molecular mechanisms of flower formation and fertility in Lycium.
METHODS AND RESULTS
In this study, 11 candidate reference genes in Lycium flower development were selected from transcriptome sequence data and evaluated with five traditional housekeeping genes from previous studies based on qRT-PCR amplification. Comparing the expression stability result of 16 candidate genes using GeNorm, NormFinder, BestKeeper, and Delta Ct algorithms, Lba04g01649 and Lba12g02820 were validated as the optimal reference genes for the flower development of Lycium.
CONCLUSIONS
The reference genes identified in this study would improve the accuracy of qRT-PCR quantification of target gene expression in Lycium flower development and facilitate future functional genomics studies on flower development. This research could lay the foundation for the study of the reproduction and development of the Lycium flower.
背景
准确且稳定表达的参照基因是成功进行实时定量 PCR（qRT-PCR）的前提。研究花发育过程中的基因表达谱可以加深我们对枸杞花形成和育性分子机制的理解。
方法和结果
本研究从转录组序列数据中选择了 11 个枸杞花发育的候选参照基因，并基于 qRT-PCR 扩增，结合之前研究中的 5 个传统管家基因对其进行了评估。使用 GeNorm、NormFinder、BestKeeper 和 Delta Ct 算法比较 16 个候选基因的表达稳定性结果，验证 Lba04g01649 和 Lba12g02820 是枸杞花发育的最优参照基因。
结论
本研究中鉴定的参照基因将提高枸杞花发育中目标基因表达的 qRT-PCR 定量准确性，并有助于未来对花发育的功能基因组学研究。本研究可为枸杞花的繁殖和发育研究奠定基础。
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