MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River, College of Plant Sciences and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730030, China.
Genes (Basel). 2021 Mar 23;12(3):459. doi: 10.3390/genes12030459.
Fr. Schmidt is a traditional medicinal herb in southwestern China and has multiple pharmacological effects on bruise injuries and rheumatoid arthritis (RA). A new drug with a good curative effect on RA has recently been developed from the extract of rhizomes, of which the main medicinal ingredients are triterpenoid saponins. Due to excessive exploitation, the wild population has been scarce and endangered in a few of its natural habitats and research on the cultivation of the plant commenced. Studies on the gene expressions related to the biosynthesis of triterpenoid saponins are not only helpful for understanding the effects of environmental factors on the medicinal ingredient accumulations but also necessary for monitoring the herb quality of the cultivated plants. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) as a sensitive and powerful technique has been widely used to detect gene expression across tissues in plants at different stages; however, its accuracy and reliability depend largely on the reference gene selection. In this study, the expressions of 10 candidate reference genes were evaluated in various organs of the wild and cultivated plants at different stages, using the algorithms of geNorm, NormFinder and BestKeeper, respectively. The purpose of this study was to identify the suitable reference genes for RT-qPCR detection in . The results showed that two reference genes were sufficient for RT-qPCR data normalization in . and can be used as suitable reference genes in most organs at various stages because of their expression stabilitywhereas the and genes were desirable in the rhizomes of the plant at the vegetative stage.
施密特神父是中国西南地区的一种传统药用植物，对跌打损伤和类风湿关节炎（RA）有多种药理作用。最近，从根茎提取物中开发出一种治疗 RA 效果良好的新药，其主要药用成分为三萜皂苷。由于过度开发，该植物在其几个自然栖息地的野生种群已经稀少，处于濒危状态，因此开始研究其栽培。研究与三萜皂苷生物合成相关的基因表达不仅有助于了解环境因素对药用成分积累的影响，而且对于监测栽培植物的草药质量也是必要的。反转录定量聚合酶链反应（RT-qPCR）作为一种敏感而强大的技术，已广泛用于检测不同阶段植物不同组织中的基因表达；然而，其准确性和可靠性在很大程度上取决于参考基因的选择。在这项研究中，使用 geNorm、NormFinder 和 BestKeeper 算法分别评估了 10 个候选参考基因在野生和栽培植物不同阶段各种器官中的表达情况。本研究的目的是鉴定适合 RT-qPCR 检测的参考基因。结果表明，对于 ，使用两个参考基因即可对 RT-qPCR 数据进行归一化。 和 在大多数器官的各个阶段都具有稳定的表达，因此可以作为合适的参考基因；而在植物营养阶段的根茎中， 和 基因则是理想的参考基因。
