Selection of Suitable Reference Genes for Gene Expression Normalization Studies in Dendrobium huoshanense

Area of Science:

• Molecular Biology
• Plant Science
• Genetics

Background:

• Dendrobium huoshanense is a precious herb with significant medicinal and edible value in China.
• Accurate gene expression analysis using quantitative real-time polymerase chain reaction (qPCR) is crucial for understanding its biosynthetic pathways.
• No validated reference genes have been previously reported for D. huoshanense, hindering reliable gene expression studies.

Purpose of the Study:

• To identify and evaluate the expression stability of candidate reference genes in D. huoshanense.
• To provide reliable reference genes for accurate gene expression normalization in D. huoshanense under various experimental conditions.

Main Methods:

• Fifteen candidate reference genes were selected and their expression stability was assessed in different tissues, under abiotic stresses, and hormone treatments.
• Expression stability was analyzed using geNorm, NormFinder, and BestKeeper statistical programs.
• The RefFinder program was used for comprehensive validation and ranking of the selected reference genes.

Main Results:

• The expression stability of 15 candidate reference genes was evaluated across diverse experimental conditions.
• TBP, NCBP2, and CYP71 were identified as the top three most stable reference genes.
• These validated reference genes enable accurate normalization of gene expression data in D. huoshanense.

Conclusions:

• This study provides the first comprehensive evaluation and selection of reference genes for D. huoshanense.
• TBP, NCBP2, and CYP71 are recommended for reliable gene expression normalization in future research.
• These findings will significantly benefit molecular studies on D. huoshanense, particularly concerning its medicinal properties.