香芋是一种广泛使用的淀粉资源植物。使用实时定量聚合酶链反应(RT-qPCR)定量与芋生长相关的功能基因的表达水平至关重要。然而,获得可靠的RT-qPCR结果,数据归一化需要适当的参考基因(RGs)。在这项研究中,我们使用来自芋的转录组数据集筛选了七个新的候选RGs,包含来自生长球茎和各种组织的数据。这七个新的RGs的表达稳定性,以及常用的RGs肌动蛋白,EF1-α,和β-微管蛋白,使用DeltaCT进行评估,BestKeeper,geNorm,和NormFinder算法。此外,我们使用RefFinder程序进行了全面分析,并使用目标基因验证了结果,CeAGPL1.研究结果表明,ACY-1和PIA2是球茎生长过程中归一化的最佳多个RGs,而COX10和Armc8适用于包括各种类型组织的样品。此外,我们发现了三个RGs,Armc8,COX10和CCX4L,是干旱胁迫的最佳RGs。这项研究首次评估了芋中RGs的适用性。确定的RGs为研究球茎生长提供了宝贵的资源,不同的组织,和干旱胁迫。这项研究有助于提高我们对控制芋生长的潜在机制的理解。
Taro is a widely utilized starch resource plant. It is essential to quantify the expression levels of functional genes associated with taro growth using real-time quantitative polymerase chain reaction (RT-qPCR). However, to obtain reliable RT-qPCR results, appropriate reference genes (RGs) are required for data normalization. In this study, we screened seven novel candidate RGs using transcriptome datasets from taro, encompassing data from growth corms and various tissues. The expression stability of these seven new RGs, along with the commonly used RGs Actin, EF1-α, and β-tubulin, was assessed using Delta Ct, BestKeeper, geNorm, and NormFinder algorithms. Furthermore, we conducted a comprehensive analysis using the RefFinder program and validated the results using the target gene, CeAGPL1. The findings revealed that ACY-1 and PIA2 were the optimal multiple RGs for normalization during corm growth, while COX10 and Armc8 were suitable for samples including various types of tissues. Furthermore, we found three RGs, Armc8, COX10 and CCX4L, were the optimal RGs for drought stress. This study assessed the suitability of RGs in taro for the first time. The identified RGs provide valuable resources for studying corm growth, diverse tissues, and drought stress. This study contributes to the advancement of our understanding of the underlying mechanisms that govern the growth of taro.