Validation of reference genes for quantitative gene expression analysis in Auricularia cornea

Area of Science:

• Mycology
• Molecular Biology
• Biotechnology

Background:

• Auricularia cornea (also known as A. polytricha) is a significant cultivated mushroom species, particularly in China.
• Accurate gene expression studies require validated reference genes for normalization.
• Previous reference gene selection for A. cornea relied on data from other species, lacking species-specific validation.

Purpose of the Study:

• To systematically evaluate the stability of nine candidate reference genes in Auricularia cornea.
• To identify the most suitable reference genes for accurate gene expression analysis in this species.

Main Methods:

• Nine candidate reference genes (TUBA1A, Btu, Pgm, Act1, PP2A, UBQ, Gapdh, 18S, 28S) were selected.
• Gene expression stability was analyzed across different strains and developmental stages.
• Four algorithms (geNorm, NormFinder, BestKeeper, RefFinder) were employed for comprehensive evaluation.

Main Results:

• Polyubiquitin (UBQ) exhibited the highest stability, while 18S ribosomal protein (18S) was the least stable.
• Only two genes, protein phosphatase 2A regulatory subunit (PP2A) and UBQ, were deemed adequate for use as reference genes.
• The overall suitability of the evaluated candidate genes was limited.

Conclusions:

• This study provides the first systematic validation of reference genes for Auricularia cornea.
• Protein phosphatase 2A regulatory subunit (PP2A) and Polyubiquitin (UBQ) are recommended as reliable internal control genes for future research in A. cornea.
• The findings establish a foundation for precise gene expression quantification in this important mushroom species.