Zhang Lu, Cai Yanfei, Zhang Mingchao, Du Guanghui, Wang Jihua
Flower Research Institute of Yunnan Academy of Agricultural Sciences, Kunming, China.
National Engineering Research Center for Ornamental Horticulture, Kunming, China.
Front Genet. 2022 Apr 14;13:876482. doi: 10.3389/fgene.2022.876482. eCollection 2022.
There has been no systematic identification and screening of candidate reference genes for normalization of quantitative real-time PCR (qRT-PCR) results in to date. Therefore, the present study used and Ubiquitin to predict their stabilities on different aboveground tissues (matured leaves (ML), stem tips (STM), and flower buds (FB)) at different developmental stages (young and adult plants) using five statistical algorithms: Delta Ct method, BestKeeper, geNorm, Normfinder, and RefFinder. The findings were confirmed using ML obtained from plants that had been stressed by drought. By using RefFinder with ML samples collected under drought conditions, it was determined that the top five most stable reference genes were , whereas the least stable reference gene was Ubiquitin. In addition, under control conditions, were selected as the highest stable potential reference genes at the juvenile stage of with ML and STM. When ML and STM were combined with drought-stressed samples, , or their combination proved to be the most effective qRT-PCR primers. The findings will aid in the improvement of the precision and reliability of qRT-PCR data and laying the groundwork for future gene functional studies in .
迄今为止,尚未对用于定量实时PCR(qRT-PCR)结果标准化的候选参考基因进行系统的鉴定和筛选。因此,本研究使用[基因名称]和泛素,运用Delta Ct法、BestKeeper、geNorm、Normfinder和RefFinder这五种统计算法,预测它们在不同发育阶段(幼龄和成年植株)的不同地上组织(成熟叶片(ML)、茎尖(STM)和花芽(FB))中的稳定性。通过对受干旱胁迫的植株获取的ML进行验证,证实了这些发现。使用RefFinder对干旱条件下收集的ML样本进行分析,确定了最稳定的前五个参考基因是[基因名称],而最不稳定的参考基因是泛素。此外,在对照条件下,[基因名称]被选为[植物名称]幼龄期ML和STM中最稳定的潜在参考基因。当将ML和STM与干旱胁迫样本相结合时,[基因名称]或它们的组合被证明是最有效的qRT-PCR引物。这些发现将有助于提高qRT-PCR数据的准确性和可靠性,并为[植物名称]未来的基因功能研究奠定基础。