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

被引:5
|
作者
Chen, Xingzhou [1 ,2 ,3 ]
Chen, Xinggang [1 ,2 ,3 ]
Tan, Qian [1 ,2 ,3 ]
He, Yuan [1 ,2 ,3 ]
Wang, Zhikai [1 ,2 ,3 ]
Zhou, Guoying [1 ,2 ,3 ]
Liu, Junang [1 ,2 ,3 ]
机构
[1] Cent South Univ Forestry & Technol, Key Lab Cultivat & Protect Nonwood Forest Trees, Minist Educ, Changsha, Peoples R China
[2] Cent South Univ Forestry & Technol, Key Lab Natl Forestry & Grassland Adm Control Arti, Changsha, Peoples R China
[3] Cent South Univ Forestry & Technol, Hunan Prov Key Lab Control Forest Dis & Pests, Changsha, Peoples R China
基金
中国国家自然科学基金;
关键词
Colletotrichum fructicola; RT-qPCR; Camellia oleifera; reference gene; transcriptome; TIME-QUANTITATIVE PCR; NORMALIZATION; VALIDATION; EXPRESSION; INFORMATION;
D O I
10.3389/fmicb.2022.982748
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
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页数:9
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