Selection of potential reference genes for RT-qPCR in the plant pathogenic fungus Colletotrichum fructicola

Colletotrichum is widespread, and these pathogenic fungi can cause various plant diseases. Studies have shown that Colletotrichum fructicola cause oil-tea ( Camellia oleifera ) anthracnose and is widely distributed as a dominant fungus in all Ca. oleifera -producing regions. Real-time quantitative PCR(RT-qPCR) is considered the most reliable technique for simultaneously measuring relative gene expression levels in different tissues. Target genes are typically quantified using RT-qPCR to explore gene function, and reliable RT-qPCR results require data normalization using stable reference genes. No studies have reported a suitable reference gene in C. fructicola . This study has eight candidate reference genes ( CfCk , CfRpp , CfUce , CfRrp , CfAdrh , CfDd , CfAct, and CfTub ) which were selected from C. fructicola - Ca. oleifera transcriptome data and evaluated and sequenced using geNorm, NormFinder, and BestKeeper algorithms. The results showed that CfRrp had better stability in C. fructicola , both during the growth of pure pathogenic fungi and during the invasion of different oil-tea leaves. After normalization with CfRrp , the differentially expressed target genes were similar to the transcriptome. Our work provides suitable reference genes for future studies to quantify target gene expression levels in C. fructicola.