Lv Yao, Li Yanyan, Liu Xiaohui, Xu Kun
College of Horticulture Science and Engineering, Shandong Agricultural University, Tai'an, China.
Collaborative Innovation Center of Fruit & Vegetable Quality and Efficient Production in Shandong, Tai'an, China.
Front Genet. 2020 Nov 3;11:586098. doi: 10.3389/fgene.2020.586098. eCollection 2020.
Quantitative real-time PCR (qRT-PCR) is widely used in the detection of gene expression level. However, there is no suitable ginger reference gene for qPCR analysis. Therefore, it is the primary task to select and validate the appropriate ginger reference gene to normalize the expression of target genes. In this study, 14 candidate reference genes were selected and analyzed in different tissues (leaf, and rhizome), different development stages, different varieties, and abiotic stress (ABA and salt stress). Expression stability was calculated using geNorm and NormFinder, Bestkeeper, and RefFinder. For abiotic stress and total conditions, 28S and COX were identified as the most stable genes. In addition, RPII was the most stable in the different development stages and different varieties. TEF2 and RPL2 were the least stably expressed in the tissue and all the conditions. In order to verify the feasibility of these genes as reference genes, we used the most stable and least stable reference genes to normalize the expression levels of genes under different conditions. This work can provide theoretical support for future research on ginger gene expression.
定量实时聚合酶链反应(qRT-PCR)广泛应用于基因表达水平的检测。然而,目前尚无适用于生姜qPCR分析的内参基因。因此,选择并验证合适的生姜内参基因以标准化目标基因的表达是首要任务。本研究选取了14个候选内参基因,在不同组织(叶片和根茎)、不同发育阶段、不同品种以及非生物胁迫(脱落酸和盐胁迫)条件下进行分析。使用geNorm、NormFinder、Bestkeeper和RefFinder计算表达稳定性。对于非生物胁迫和总体条件,28S和COX被确定为最稳定的基因。此外,RPII在不同发育阶段和不同品种中最稳定。TEF2和RPL2在组织和所有条件下表达最不稳定。为了验证这些基因作为内参基因的可行性,我们使用最稳定和最不稳定的内参基因来标准化不同条件下基因的表达水平。本研究可为今后生姜基因表达研究提供理论支持。