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

被引:7
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
收藏
页数:9
相关论文
共 34 条
[21]   Transcription Factor CfSte12 of Colletotrichum fructicola Is a Key Regulator of Early Apple Glomerella Leaf Spot Pathogenesis [J].
Liu, Wenkui ;
Liang, Xiaofei ;
Gleason, Mark L. ;
Cao, Mengyu ;
Zhang, Rong ;
Sun, Guangyu .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2020, 87 (01) :1-17
[22]   Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method [J].
Livak, KJ ;
Schmittgen, TD .
METHODS, 2001, 25 (04) :402-408
[23]   Innovative tools for detection of plant pathogenic viruses and bacteria [J].
López, MM ;
Bertolini, E ;
Olmos, A ;
Caruso, P ;
Gorris, MT ;
Llop, P ;
Penyalver, R ;
Cambra, M .
INTERNATIONAL MICROBIOLOGY, 2003, 6 (04) :233-243
[24]   Comprehensive selection of reference genes for quantitative gene expression analysis during seed development in Brassica napus [J].
Machado, Ronei Dorneles ;
Christoff, Ana Paula ;
Loss-Morais, Guilherme ;
Margis-Pinheiro, Marcia ;
Margis, Rogerio ;
Koerbes, Ana Paula .
PLANT CELL REPORTS, 2015, 34 (07) :1139-1149
[25]   RRS1 and RPS4 provide a dual Resistance-gene system against fungal and bacterial pathogens [J].
Narusaka, Mari ;
Shirasu, Ken ;
Noutoshi, Yoshiteru ;
Kubo, Yasuyuki ;
Shiraishi, Tomonori ;
Iwabuchi, Masaki ;
Narusaka, Yoshihiro .
PLANT JOURNAL, 2009, 60 (02) :218-226
[26]   Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper - Excel-based tool using pair-wise correlations [J].
Pfaffl, MW ;
Tichopad, A ;
Prgomet, C ;
Neuvians, TP .
BIOTECHNOLOGY LETTERS, 2004, 26 (06) :509-515
[27]   Transcriptome-based identification and validation of reference genes for plant-bacteria interaction studies using Nicotiana benthamiana [J].
Pombo, Marina A. ;
Ramos, Romina N. ;
Zheng, Yi ;
Fei, Zhangjun ;
Martin, Gregory B. ;
Rosli, Hernan G. .
SCIENTIFIC REPORTS, 2019, 9 (1)
[28]   Spatial distribution of Metarhizium clade 1 in agricultural landscapes with arable land and different semi-natural habitats [J].
Schneider, S. ;
Widmer, F. ;
Jacot, K. ;
Koelliker, R. ;
Enkerli, J. .
APPLIED SOIL ECOLOGY, 2012, 52 :20-28
[29]   A novel effector CfEC92 of Colletotrichum fructicola contributes to glomerella leaf spot virulence by suppressing plant defences at the early infection phase [J].
Shang Shengping ;
Wang Bo ;
Zhang Song ;
Liu Guangli ;
Liang Xiaofei ;
Zhang Rong ;
Gleason, Mark L. ;
Sun Guangyu .
MOLECULAR PLANT PATHOLOGY, 2020, 21 (07) :936-950
[30]   Gene information for fungal plant pathogens from expressed sequences [J].
Skinner, W ;
Keon, J ;
Hargreaves, J .
CURRENT OPINION IN MICROBIOLOGY, 2001, 4 (04) :381-386