AUTHOR=Chen Xingzhou , Chen Xinggang , Tan Qian , He Yuan , Wang Zhikai , Zhou Guoying , Liu Junang 

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

JOURNAL=Frontiers in Microbiology

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2022.982748

DOI=10.3389/fmicb.2022.982748

ISSN=1664-302X

ABSTRACT=<p><italic>Colletotrichum</italic> is widespread, and these pathogenic fungi can cause various plant diseases. Studies have shown that <italic>Colletotrichum fructicola</italic> cause oil-tea (<italic>Camellia oleifera</italic>) anthracnose and is widely distributed as a dominant fungus in all <italic>Ca. oleifera</italic>-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 <italic>C. fructicola</italic>. This study has eight candidate reference genes (<italic>CfCk</italic>, <italic>CfRpp</italic>, <italic>CfUce</italic>, <italic>CfRrp</italic>, <italic>CfAdrh</italic>, <italic>CfDd</italic>, <italic>CfAct,</italic> and <italic>CfTub</italic>) which were selected from <italic>C. fructicola</italic>-<italic>Ca. oleifera</italic> transcriptome data and evaluated and sequenced using geNorm, NormFinder, and BestKeeper algorithms. The results showed that <italic>CfRrp</italic> had better stability in <italic>C. fructicola</italic>, both during the growth of pure pathogenic fungi and during the invasion of different oil-tea leaves. After normalization with <italic>CfRrp</italic>, 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 <italic>C. fructicola.</italic></p>