title: Selection of reference genes for gene expression studies in Siberian Apricot (Prunus sibirica L.) Germplasm using quantitative real-time PCR.
abstract: Quantitative real time reverse transcription polymerase chain reaction has been applied in a vast range of studies of gene expression analysis. However, real-time PCR data must be normalized with one or more reference genes. In this study, eleven putative consistently expressed genes (ACT, TUA, TUB, CYP, DNAj, ELFA, F-box27, RPL12, GAPDH, UBC and UBQ) in nine Siberian Apricot Germplasms (including much variability) were evaluated for their potential as references for the normalization of gene expression by NormFinder and geNorm programs. From our studies, ACT, UBC, CYP, UBQ and RPL12 as suitable for normalization were identified by geNorm, while UBC and CYP as the best pair by NormFinder. Moreover, UBC was selected as the most stably expressed gene by both algorithms in different Siberian Apricot seed samples. We also detected that a set of three genes (ACT, CYP and UBC) by geNorm as control for normalization could lead to accurate results. Furthermore, the expression levels of oleosin gene were analyzed to validate the suitability of the selected reference genes. These obtained experimental results could make an important contribution to normalize real-time PCR data for gene expression analysis in Siberian Apricot Germplasm.
authors.affiliation: College of Biological Sciences and Biotechnology, College of Nature Conservation, National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry University, Beijing, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Niu
authors.affiliation: College of Biological Sciences and Biotechnology, College of Nature Conservation, National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry University, Beijing, China.
authors.forename: Baoqing
authors.initials: B
authors.lastname: Zhu
authors.affiliation: College of Biological Sciences and Biotechnology, College of Nature Conservation, National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry University, Beijing, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Cai
authors.affiliation: Jigongshan National Nature Reserve, Xingyang, China.
authors.forename: Peixue
authors.initials: P
authors.lastname: Li
authors.affiliation: Research Institute of Forestry, Chinese Academy of Forestry, Beijing, China.
authors.forename: Libing
authors.initials: L
authors.lastname: Wang
authors.affiliation: Jigongshan National Nature Reserve, Xingyang, China.
authors.forename: Huitang
authors.initials: H
authors.lastname: Dai
authors.affiliation: Jigongshan National Nature Reserve, Xingyang, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Qiu
authors.affiliation: Research Institute of Forestry, Chinese Academy of Forestry, Beijing, China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Yu
authors.affiliation: Jigongshan National Nature Reserve, Xingyang, China.
authors.forename: Denglong
authors.initials: D
authors.lastname: Ha
authors.affiliation: Jigongshan National Nature Reserve, Xingyang, China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Zhao
authors.affiliation: College of Biological Sciences and Biotechnology, College of Nature Conservation, National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry University, Beijing, China.
authors.forename: Zhixiang
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: College of Biological Sciences and Biotechnology, College of Nature Conservation, National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry University, Beijing, China.
authors.forename: Shanzhi
authors.initials: S
authors.lastname: Lin
mesh_terms: D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D017343:Genes, Plant / Q000235:genetics*; D027861:Prunus / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary
references.citation: BMC Mol Biol. 2009 Jul 20;10:71
references.pmid: 19619301
references.citation: BMC Plant Biol. 2004 Aug 18;4:14
references.pmid: 15317655
references.citation: Reprod Domest Anim. 2012 Dec;47(6):e88-91
references.pmid: 23289123
references.citation: Anal Biochem. 2009 Apr 15;387(2):238-42
references.pmid: 19454243
references.citation: Anal Biochem. 2002 Oct 15;309(2):293-300
references.pmid: 12413463
references.citation: J Exp Bot. 2005 Nov;56(421):2907-14
references.pmid: 16188960
references.citation: Biotechniques. 2000 Aug;29(2):332-7
references.pmid: 10948434
references.citation: J Biotechnol. 1999 Oct 8;75(2-3):291-5
references.pmid: 10617337
references.citation: BMC Mol Biol. 2010 Jan 20;11:8
references.pmid: 20089196
references.citation: J Exp Bot. 2005 Jul;56(417):1877-86
references.pmid: 15911563
references.citation: Biochem Biophys Res Commun. 2006 Jun 30;345(2):646-51
references.pmid: 16690022
references.citation: Plant Cell Rep. 2013 May;32(5):715-29
references.pmid: 23529547
references.citation: Cancer Res. 2004 Aug 1;64(15):5245-50
references.pmid: 15289330
references.citation: Clin Chem. 2009 Apr;55(4):611-22
references.pmid: 19246619
references.citation: Fish Shellfish Immunol. 2013 Mar;34(3):939-45
references.pmid: 23357023
references.citation: Genome Biol. 2002 Jun 18;3(7):RESEARCH0034
references.pmid: 12184808
references.citation: World J Microbiol Biotechnol. 2013 Sep;29(9):1563-71
references.pmid: 23508400
references.citation: Mol Biotechnol. 2014 Mar;56(3):210-22
references.pmid: 24037409
references.citation: Ecotoxicology. 2014 Apr;23(3):459-71
references.pmid: 24566730
references.citation: BMC Plant Biol. 2010 Jan 07;10:4
references.pmid: 20056000
references.citation: PLoS One. 2012;7(3):e33278
references.pmid: 22479379
references.citation: BMC Plant Biol. 2010 Mar 21;10:49
references.pmid: 20302670
references.citation: Vet Immunol Immunopathol. 2008 Dec 15;126(3-4):392-5
references.pmid: 18824265
references.citation: Genes Immun. 2005 Jun;6(4):279-84
references.pmid: 15815687
references.citation: Biotechnol Lett. 2004 Mar;26(6):509-15
references.pmid: 15127793
references.citation: Methods. 2010 Apr;50(4):262-70
references.pmid: 20060046
references.citation: J Mol Endocrinol. 2005 Jun;34(3):597-601
references.pmid: 15956331
references.citation: Biochem Biophys Res Commun. 2012 Jul 20;424(1):118-23
references.pmid: 22732401
references.citation: BMC Mol Biol. 2008 Jun 23;9:59
references.pmid: 18573215
references.citation: Plant Signal Behav. 2010 Apr;5(4):447-9
references.pmid: 20081376
references.citation: J Mol Endocrinol. 2002 Aug;29(1):23-39
references.pmid: 12200227
references.citation: Food Chem Toxicol. 2011 Dec;49(12):3136-41
references.pmid: 21896302
references.citation: Neurosci Lett. 2003 Mar 13;339(1):62-6
references.pmid: 12618301
references.citation: BMC Mol Biol. 2010 Sep 21;11:74
references.pmid: 20858237