College of Forestry, Northwest A&F University, Yangling 712100, China.
Molecules. 2018 Mar 30;23(4):802. doi: 10.3390/molecules23040802.
Real-time reverse transcription quantitative PCR has become a common method for studying gene expression, however, the optimal selection of stable reference genes is a prerequisite for obtaining accurate quantification of transcript abundance. Suitable reference genes for RT-qPCR have not yet been identified for Chinese prickly ash ( Maxim.). Chinese prickly ash is the source of an important food seasoning in China. In recent years, Chinese prickly ash has also been developed as a medicinal plant. The expression stabilities of ten genes (, , , , , , , , , and ) were evaluated in roots, stems, leaves, flowers and fruits at five developmental stages and also under stress from cold, drought, and salt. To do this we used three different statistical algorithms: geNorm, NormFinder and BestKeeper. Among the genes investigated, and were found to be most stable for the different cultivars and different tissues examined, and for fruit developmental stage. Meanwhile, and were most stable under cold treatment, and under drought treatment and and under salt treatment. and for all samples evaluated were most stably expressed, but , and were found to be generally unreliable as reference genes. Our results provide a basis for the future selection of reference genes for biological research with Chinese prickly ash, under a variety of conditions.
实时逆转录定量 PCR 已成为研究基因表达的常用方法,然而,选择稳定的参考基因是获得转录丰度准确定量的前提。对于花椒( Maxim.),尚未确定适合 RT-qPCR 的参考基因。花椒是中国重要的调味料来源。近年来,花椒也被开发为药用植物。在五个发育阶段和冷、旱、盐胁迫下,评估了根、茎、叶、花和果实中 10 个基因( 、 、 、 、 、 、 、 和 )的表达稳定性。为此,我们使用了三种不同的统计算法:geNorm、NormFinder 和 BestKeeper。在所研究的基因中, 和 被发现对于不同品种和不同组织是最稳定的, 和 对于果实发育阶段是最稳定的。同时, 和 在冷处理下最稳定, 和 在干旱处理下最稳定, 和 在盐处理下最稳定。 和 在所有评估的样本中表达最稳定,但 、 和 被发现作为参考基因通常不可靠。我们的研究结果为未来在各种条件下进行花椒的生物学研究选择参考基因提供了依据。