# qPCR-Based Reference Gene Validation in Canarium album: Stability Across Varieties and Developmental Stages

## TL;DR

This study identifies RPN2B and NIFS1 as stable reference genes for gene expression studies in Chinese olive fruits across varieties and developmental stages.

## Contribution

The study validates and recommends RPN2B and NIFS1 as reliable reference genes for Chinese olive gene expression analysis.

## Key findings

- RPN2B and NIFS1 were consistently ranked as the most stable genes across multiple algorithms.
- The study evaluated eight candidate genes across 33 fruit samples from different varieties and developmental stages.
- Four methods (ΔCq, geNorm, NormFinder, BestKeeper) were used to assess gene stability.

## Abstract

To obtain stable Chinese olive reference genes, eight genes (RPN2B, PIP1.4, NIFS1, RPS16, At5g12110, HSC-2, ABCG44, LOS1) exhibiting stable expression were identified as candidate reference genes from the transcriptome. The expression stability of these genes was evaluated across 33 Chinese olive fruit samples from different varieties and seven developmental stages. The most stable reference genes were determined through comparisons using ΔCq, geNorm, NormFinder, BestKeeper, and RefFinder. Analysis revealed that RPN2B and NIFS1 were consistently ranked among the most stable genes across the different algorithms and exhibited stable expression. Therefore, they are recommended as suitable reference genes for gene expression studies in Chinese olive fruits across different varieties and developmental stages. The four different methods of reference gene stability analysis were used to identify the most stable reference genes in different varieties and developmental stages of Chinese olive fruits, which can be used as a reference for the selection of reference genes in the subsequent gene expression studies of Chinese olive fruits.