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基于转录组的芋(Colocasia esculenta)球茎生长阶段、不同组织和干旱胁迫参考基因的鉴定和验证。

Transcriptome-based identification and validation of reference genes for corm growth stages, different tissues, and drought stress in Taro (Colocasia esculenta).

机构信息

Vegetable Research Institute, Guangxi Zhuang Autonomous Region Academy of Agricultural Sciences, Nanning, 530007, China.

New Technology Entrepreneur Center, Nanning, 530007, China.

出版信息

BMC Plant Biol. 2024 May 30;24(1):478. doi: 10.1186/s12870-024-05199-x.

Abstract

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.

摘要

芋艿是一种广泛应用的淀粉资源植物。利用实时定量聚合酶链反应(RT-qPCR)定量测定与芋艿生长相关的功能基因的表达水平至关重要。然而,为了获得可靠的 RT-qPCR 结果,需要合适的内参基因(RGs)进行数据归一化。本研究利用芋艿生长球茎和各种组织的转录组数据集,筛选了 7 个新的候选 RG。采用 Delta Ct、BestKeeper、geNorm 和 NormFinder 算法评估了这 7 个新 RG 以及常用 RG Actin、EF1-α和β-微管蛋白的表达稳定性。此外,我们使用 RefFinder 程序进行了全面分析,并使用靶基因 CeAGPL1 验证了结果。结果表明,ACY-1 和 PIA2 是球茎生长过程中归一化的最佳多 RG,而 COX10 和 Armc8 适合包括各种组织类型的样本。此外,我们发现三个 RG,Armc8、COX10 和 CCX4L,是干旱胁迫的最佳 RG。本研究首次评估了 RG 在芋艿中的适用性。鉴定的 RG 为研究球茎生长、多种组织和干旱胁迫提供了有价值的资源。本研究有助于我们深入了解芋艿生长的调控机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b737/11137888/897a83d5bec7/12870_2024_5199_Fig1_HTML.jpg

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