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International Journal of Medicinal Mushrooms
Impact-faktor: 1.423 5-jähriger Impact-Faktor: 1.525 SJR: 0.431 SNIP: 0.661 CiteScore™: 1.38

ISSN Druckformat: 1521-9437
ISSN Online: 1940-4344

Volumes:
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International Journal of Medicinal Mushrooms

DOI: 10.1615/IntJMedMushrooms.v18.i2.70
pages 165-175

Selection and Validation of Reference Genes for Quantitative Real-Time Polymerase Chain Reaction Studies in Mossy Maze Polypore, Cerrena unicolor (Higher Basidiomycetes)

Jie Yang
College of Biological Sciences and Technology, Fuzhou University, Fuzhou, Fujian, China; Fujian Key Laboratory of Marine Enzyme Engineering, Fuzhou, Fujian, China
Qi Lin
College of Biological Sciences and Technology, Fuzhou University, Fuzhou, Fujian, China
Juan Lin
College of Biological Sciences and Technology, Fuzhou University, Fuzhou, Fujian, China; Fujian Key Laboratory of Marine Enzyme Engineering, Fuzhou, Fujian, China
Xiuyun Ye
College of Biological Sciences and Technology, Fuzhou University, Fuzhou, Fujian, China; Fujian Key Laboratory of Marine Enzyme Engineering, Fuzhou, Fujian, China

ABSTRAKT

With its ability to produce ligninolytic enzymes such as laccases, white-rot basidiomycete Cerrena unicolor, a medicinal mushroom, has great potential in biotechnology. Elucidation of the expression profiles of genes encoding ligninolytic enzymes are important for increasing their production. Quantitative real-time polymerase chain reaction (qPCR) is a powerful tool to study transcriptional regulation of genes of interest. To ensure accuracy and reliability of qPCR analysis of C. unicolor, expression levels of seven candidate reference genes were studied at different growth phases, under various induction conditions, and with a range of carbon/nitrogen ratios and carbon and nitrogen sources. The stability of the genes were analyzed with five statistical approaches, namely geNorm, NormFinder, BestKeeper, the ΔCt method, and RefFinder. Our results indicated that the selection of reference genes varied with sample sets. A combination of four reference genes (Cyt-c, ATP6, TEF1, and β-tubulin) were recommended for normalizing gene expression at different growth phases. GAPDH and Cyt-c were the appropriate reference genes under different induction conditions. ATP6 and TEF1 were most stable in fermentation media with various carbon/nitrogen ratios. In the fermentation media with various carbon or nitrogen sources, 18S rRNA and GAPDH were the references of choice. The present study represents the first validation analysis of reference genes in C. unicolor and serves as a foundation for its qPCR analysis.


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