Identification of Ginger (Zingiber officinale Roscoe) Reference Genes for Gene Expression Analysis

Area of Science:

• Plant Molecular Biology
• Genomics
• Biotechnology

Background:

• Quantitative real-time PCR (qRT-PCR) is a standard method for measuring gene expression levels.
• Accurate normalization requires stable reference genes, which are currently lacking for ginger (Zingiber officinale).
• Identifying suitable reference genes is essential for reliable gene expression studies in ginger.

Purpose of the Study:

• To identify and validate stable reference genes for qRT-PCR in ginger across diverse experimental conditions.
• To evaluate the expression stability of 14 candidate reference genes.
• To provide a reliable basis for normalizing target gene expression in ginger.

Main Methods:

• Analysis of 14 candidate reference genes in different ginger tissues, developmental stages, varieties, and under abiotic stress (ABA, salt).
• Expression stability assessment using geNorm, NormFinder, Bestkeeper, and RefFinder algorithms.
• Validation of selected reference genes by normalizing the expression of *ZoSPS* genes.

Main Results:

• Genes 28S and COX demonstrated the highest expression stability under abiotic stress and overall conditions.
• RPII was the most stable reference gene for different developmental stages and varieties.
• TEF2 and RPL2 were identified as the least stable reference genes.
• The choice of reference gene significantly impacted the normalization of *ZoSPS* gene expression.

Conclusions:

• 28S and COX are recommended as reliable reference genes for ginger under abiotic stress and general conditions.
• RPII is suitable for normalizing gene expression in different developmental stages and varieties of ginger.
• The findings provide critical support for accurate gene expression studies in ginger, advancing research in plant science and breeding.