Rodríguez-Parra Alba, Picazo-Aragonés Jesús, Balao Francisco
Departament of Plant Biology and Ecology, Faculty of Biology, University of Seville, Apdo. 1095, 41080 Seville, Spain.
Plants (Basel). 2022 Feb 14;11(4):518. doi: 10.3390/plants11040518.
is an endemic complex which is considered the largest polyploid series within the Dianthus genus. This polyploid species involves four cytotypes (2×, 4×, 6× and 12×) with spatial and ecological segregation. The study of gene expression in polyploid species must be very rigorous because of the effects of duplications on gene regulation. In these cases, real-time polymerase chain reaction (qPCR) is the most appropriate technique for determining the gene expression profile because of its high sensitivity. The relative quantification strategy using qPCR requires genes with stable expression, known as reference genes, for normalization. In this work, we evaluated the stability of 13 candidate genes to be considered reference genes in leaf and petal tissues in . Several statistical analyses were used to determine the most stable candidate genes: Bayesian analysis, network analysis based on equivalence tests, geNorm and BestKeeper algorithms. In the leaf tissue, the most stable candidate genes were TIP41, TIF5A, PP2A and SAMDC. Similarly, the most adequate reference genes were H3.1, TIP41, TIF5A and ACT7 in the petal tissue. Therefore, we suggest that the best reference genes to compare different ploidy levels for both tissues in are TIP41 and TIF5A.
是一种地方特有复合体，被认为是石竹属中最大的多倍体系列。这个多倍体物种涉及四种细胞类型（2倍体、4倍体、6倍体和12倍体），存在空间和生态隔离。由于基因重复对基因调控的影响，对多倍体物种中的基因表达进行研究必须非常严谨。在这些情况下，实时聚合酶链反应（qPCR）因其高灵敏度而成为确定基因表达谱的最合适技术。使用qPCR的相对定量策略需要具有稳定表达的基因，即所谓的参考基因，用于标准化。在这项工作中，我们评估了13个候选基因在[具体植物名称]叶片和花瓣组织中作为参考基因的稳定性。使用了几种统计分析方法来确定最稳定的候选基因：贝叶斯分析、基于等效性检验的网络分析、geNorm和BestKeeper算法。在叶片组织中，最稳定的候选基因是TIP41、TIF5A、PP2A和SAMDC。同样，在花瓣组织中最合适的参考基因是H3.1、TIP41、TIF5A和ACT7。因此，我们建议在[具体植物名称]中，用于比较两种组织不同倍性水平的最佳参考基因是TIP41和TIF5A。
