Reference Genes for Accurate Transcript Normalization in Citrus Genotypes under Different Experimental Conditions
作者： Valéria Mafra , Karen S Kubo , Marcio Alves-Ferreira , Marcelo Ribeiro-Alves , Rodrigo M Stuart
DOI: 10.1371/JOURNAL.PONE.0031263
关键词: Gene expression profiling 、 Reference genes 、 Gene expression 、 Gene 、 Regulation of gene expression 、 Transcriptome 、 Genetics 、 Biology 、 Candidate gene 、 Biotic stress
摘要: Real-time reverse transcription PCR (RT-qPCR) has emerged as an accurate and widely used technique for expression profiling of selected genes. However, obtaining reliable measurements depends on the selection appropriate reference genes gene normalization. The aim this work was to assess stability 15 candidate determine which set is best suited transcript normalization in citrus different tissues organs leaves challenged with five pathogens (Alternaria alternata, Phytophthora parasitica, Xylella fastidiosa Candidatus Liberibacter asiaticus). We tested traditional orthologs Arabidopsis thaliana described superior based transcriptome data. geNorm NormFinder algorithms were find normalize all samples conditions tested. Additionally, each biotic stress individually analyzed by geNorm. In general, FBOX (encoding a member F-box family) GAPC2 (GAPDH) most stable assessed under subsets tested, while CYP (cyclophilin), TUB (tubulin) CtP (cathepsin) least stably expressed found. Validation suitable normalizing level WRKY70 factor infected asiaticus showed that arbitrary use without previous testing could lead misinterpretation Our results revealed FBOX, SAND (a family protein), UPL7 (ubiquitin protein ligase 7) be genes, we recommend their studies species relatives. This constitutes first systematic analysis stress.
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core.ac.uk 参考文章(52)
Yvonne J K Edwards, Elizabeth Hellen, Greg Elgar, Tim Carver, Amanda Cottage, Sunil Patel, Miriam B D Portela, Tanya Vavouri, Lisa Mullan, Molecular characterisation of the SAND protein family: a study based on comparative genomics, structural bioinformatics and phylogeny. Cellular & Molecular Biology Letters. ,vol. 9, pp. 739- 753 ,(2004)
Jo Vandesompele, Katleen De Preter, Filip Pattyn, Bruce Poppe, Nadine Van Roy, Anne De Paepe, Frank Speleman, Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes Genome Biology. ,vol. 3, pp. 1- 12 ,(2002) , 10.1186/GB-2002-3-7-RESEARCH0034
Fernanda Cruz, Samara Kalaoun, Paula Nobile, Carlos Colombo, Juliana Almeida, Leila MG Barros, Eduardo Romano, Maria Fátima Grossi-de-Sá, Maité Vaslin, Marcio Alves-Ferreira, None, Evaluation of coffee reference genes for relative expression studies by quantitative real-time RT-PCR Molecular Breeding. ,vol. 23, pp. 607- 616 ,(2009) , 10.1007/S11032-009-9259-X
Hong-xia Miao, Yong-hua Qin, Jaime A. Teixeira da Silva, Zi-xing Ye, Gui-bing Hu, Cloning and expression analysis of S-RNase homologous gene in Citrus reticulata Blanco cv. Wuzishatangju Plant Science. ,vol. 180, pp. 358- 367 ,(2011) , 10.1016/J.PLANTSCI.2010.10.012
Mukesh Jain, Aashima Nijhawan, Akhilesh K. Tyagi, Jitendra P. Khurana, Validation of housekeeping genes as internal control for studying gene expression in rice by quantitative real-time PCR Biochemical and Biophysical Research Communications. ,vol. 345, pp. 646- 651 ,(2006) , 10.1016/J.BBRC.2006.04.140
Hongjian Wan, Zhenguo Zhao, Chuntao Qian, Yihu Sui, Ahmed Abbas Malik, Jinfeng Chen, Selection of appropriate reference genes for gene expression studies by quantitative real-time polymerase chain reaction in cucumber Analytical Biochemistry. ,vol. 399, pp. 257- 261 ,(2010) , 10.1016/J.AB.2009.12.008
RAÚL ANDRÉS CERNADAS, LUCIANA RODRIGUES CAMILLO, CELSO EDUARDO BENEDETTI, Transcriptional analysis of the sweet orange interaction with the citrus canker pathogens Xanthomonas axonopodis pv. citri and Xanthomonas axonopodis pv. aurantifolii Molecular Plant Pathology. ,vol. 9, pp. 609- 631 ,(2008) , 10.1111/J.1364-3703.2008.00486.X
Tony Remans, Karen Smeets, Kelly Opdenakker, Dennis Mathijsen, Jaco Vangronsveld, Ann Cuypers, Normalisation of real-time RT-PCR gene expression measurements in Arabidopsis thaliana exposed to increased metal concentrations Planta. ,vol. 227, pp. 1343- 1349 ,(2008) , 10.1007/S00425-008-0706-4
Tania Nolan, Rebecca E Hands, Stephen A Bustin, Quantification of mRNA using real-time RT-PCR Nature Protocols. ,vol. 1, pp. 1559- 1582 ,(2006) , 10.1038/NPROT.2006.236
M. Libault, S. Thibivilliers, D. D. Bilgin, O. Radwan, M. Benitez, S. J. Clough, G. Stacey, Identification of Four Soybean Reference Genes for Gene Expression Normalization The Plant Genome. ,vol. 1, pp. 44- 54 ,(2008) , 10.3835/PLANTGENOME2008.02.0091