Evaluation of reference genes for transcript normalization in Fragaria chiloensis fruit and vegetative tissues

Area of Science:

• Plant Molecular Biology
• Gene Expression Analysis
• Biotechnology

Background:

• Quantitative real-time PCR (RT-qPCR) is a vital tool for gene expression studies.
• Reliable reference genes are essential for accurate normalization in RT-qPCR.
• Reference gene availability is limited in strawberry species, particularly *Fragaria chiloensis*.

Purpose of the Study:

• To identify and validate stable reference genes for RT-qPCR in *Fragaria chiloensis*.
• To analyze reference gene expression stability across various vegetative tissues and fruit developmental stages.
• To evaluate the impact of hormonal treatments (ABA, auxin) on reference gene expression stability.

Main Methods:

• Analyzed expression dynamics of 10 candidate reference genes in *F. chiloensis* tissues and fruits.
• Utilized geNorm, NormFinder, BestKeeper, and ΔCt algorithms for stability assessment.
• Employed RefFinder for comprehensive comparative ranking of candidate reference genes.

Main Results:

• Reference gene stability varied significantly across different tissues, fruit ripening stages, and hormonal treatments.
• Recommended gene pairs for specific conditions: *FcCHP2/FcACTIN1* (vegetative tissues), *FcDBP/FcCHP1* (fruit ripening), *FcGAPDH/FcDBP* (ABA), *FcCHP1/FcCHP2* (auxin).
• *FcDBP* emerged as the most stable single reference gene across most experimental conditions.

Conclusions:

• Provides a validated set of reference genes for RT-qPCR in *F. chiloensis*.
• Offers specific gene recommendations tailored to experimental context (tissue, fruit stage, treatment).
• Enhances the reliability and accuracy of gene expression studies in Chilean strawberry.