Evaluation and validation of reference genes for RT-qPCR normalization in different sweet potato tissues

Area of Science:

• Plant Molecular Biology
• Agricultural Science
• Biotechnology

Background:

• Sweet potato (Ipomoea batatas) is a globally significant crop with substantial economic impact.
• Limited research on biotechnological tools like RT-qPCR hinders sweet potato breeding programs.
• Accurate gene expression analysis is crucial for understanding and improving crop performance.

Purpose of the Study:

• To evaluate existing and identify novel reference genes for normalizing RT-qPCR data in sweet potato.
• To address the gap in molecular study methodologies for this agriculturally important species.
• To enhance the reliability of gene expression analyses in sweet potato.

Main Methods:

• Evaluated ten candidate reference genes, including previously validated and newly proposed genes.
• Analyzed gene expression stability across four different sweet potato tissues: fibrous root, tuberous root, stem, and leaf.
• Utilized the RefFinder algorithm to assess gene stability.

Main Results:

• IbACT, IbARF, and IbCYC were identified as the most stable reference genes with minimal expression variation across tissues.
• IbGAP, IbRPL, and IbCOX exhibited the least stability.
• The findings provide essential data for accurate RT-qPCR normalization in sweet potato.

Conclusions:

• The selection of appropriate reference genes is critical for the reliability of RT-qPCR studies in sweet potato.
• This research contributes to a better understanding of sweet potato's biological processes.
• The validated reference genes will facilitate future transcriptome analyses and crop improvement efforts.