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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(...

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Autores principales: Chen, Xingzhou, Chen, Xinggang, Tan, Qian, He, Yuan, Wang, Zhikai, Zhou, Guoying, Liu, Junang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9393503/
https://www.ncbi.nlm.nih.gov/pubmed/36003927
http://dx.doi.org/10.3389/fmicb.2022.982748
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author Chen, Xingzhou
Chen, Xinggang
Tan, Qian
He, Yuan
Wang, Zhikai
Zhou, Guoying
Liu, Junang
author_facet Chen, Xingzhou
Chen, Xinggang
Tan, Qian
He, Yuan
Wang, Zhikai
Zhou, Guoying
Liu, Junang
author_sort Chen, Xingzhou
collection PubMed
description 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.
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spelling pubmed-93935032022-08-23 Selection of potential reference genes for RT-qPCR in the plant pathogenic fungus Colletotrichum fructicola Chen, Xingzhou Chen, Xinggang Tan, Qian He, Yuan Wang, Zhikai Zhou, Guoying Liu, Junang Front Microbiol Microbiology 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. Frontiers Media S.A. 2022-08-08 /pmc/articles/PMC9393503/ /pubmed/36003927 http://dx.doi.org/10.3389/fmicb.2022.982748 Text en Copyright © 2022 Chen, Chen, Tan, He, Wang, Zhou and Liu. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Chen, Xingzhou
Chen, Xinggang
Tan, Qian
He, Yuan
Wang, Zhikai
Zhou, Guoying
Liu, Junang
Selection of potential reference genes for RT-qPCR in the plant pathogenic fungus Colletotrichum fructicola
title Selection of potential reference genes for RT-qPCR in the plant pathogenic fungus Colletotrichum fructicola
title_full Selection of potential reference genes for RT-qPCR in the plant pathogenic fungus Colletotrichum fructicola
title_fullStr Selection of potential reference genes for RT-qPCR in the plant pathogenic fungus Colletotrichum fructicola
title_full_unstemmed Selection of potential reference genes for RT-qPCR in the plant pathogenic fungus Colletotrichum fructicola
title_short Selection of potential reference genes for RT-qPCR in the plant pathogenic fungus Colletotrichum fructicola
title_sort selection of potential reference genes for rt-qpcr in the plant pathogenic fungus colletotrichum fructicola
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9393503/
https://www.ncbi.nlm.nih.gov/pubmed/36003927
http://dx.doi.org/10.3389/fmicb.2022.982748
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