College of Life Sciences, Yangtze University, Jingzhou 434025, China.
Genes (Basel). 2022 Jul 10;13(7):1227. doi: 10.3390/genes13071227.
is an important galactomannan gums source plant with characteristics of drought resistance, barren tolerance, and good adaptability. However, the underlying molecular mechanisms of the biological process are not yet fully understood. Real-time quantitative PCR (RT-qPCR) is an accurate and convenient method to quantify the gene expression level and transcription abundance of suitable reference genes. This study aimed to screen the best internal reference genes in under abiotic stresses, hormone treatments, and different tissues. Based on the transcriptome data, twelve candidate reference genes were selected, and ultimately, nine of them were further evaluated by the geNorm, NormFinder, BestKeeper, and RefFinder algorithms. These results show that ()and ()were the two most stable reference genes, and ()and , ()were the two most unstable reference genes across all samples under the given experimental conditions. Meanwhile, the most stable reference genes varied among the different groups and tissues. Therefore, this study suggests that it is better to use a specific reference gene for a particular case rather than using a common reference gene.
是一种重要的半乳甘露聚糖胶来源植物，具有耐旱、耐瘠薄和良好适应性的特点。然而，其生物学过程的潜在分子机制尚不完全清楚。实时荧光定量 PCR（RT-qPCR）是一种准确且方便的方法，可定量合适的内参基因的基因表达水平和转录丰度。本研究旨在筛选非生物胁迫、激素处理和不同组织条件下 的最佳内参基因。基于转录组数据，选择了 12 个候选内参基因，最终通过 geNorm、NormFinder、BestKeeper 和 RefFinder 算法进一步评估了其中的 9 个基因。结果表明，（）和（）是两个最稳定的内参基因，（）和 （）是所有样本在给定实验条件下最不稳定的两个内参基因。同时，不同组和组织之间最稳定的内参基因也有所不同。因此，本研究建议，最好针对特定情况使用特定的内参基因，而不是使用通用的内参基因。
