title: Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plant (Camellia sinensis (L.) O. Kuntze).
abstract: Reliable reference selection for the accurate quantification of gene expression under various experimental conditions is a crucial step in qRT-PCR normalization. To date, only a few housekeeping genes have been identified and used as reference genes in tea plant. The validity of those reference genes are not clear since their expression stabilities have not been rigorously examined. To identify more appropriate reference genes for qRT-PCR studies on tea plant, we examined the expression stability of 11 candidate reference genes from three different sources: the orthologs of Arabidopsis traditional reference genes and stably expressed genes identified from whole-genome GeneChip studies, together with three housekeeping gene commonly used in tea plant research. We evaluated the transcript levels of these genes in 94 experimental samples. The expression stabilities of these 11 genes were ranked using four different computation programs including geNorm, Normfinder, BestKeeper, and the comparative ∆CT method. Results showed that the three commonly used housekeeping genes of CsTUBULIN1, CsACINT1 and Cs18S rRNA1 together with CsUBQ1 were the most unstable genes in all sample ranking order. However, CsPTB1, CsEF1, CsSAND1, CsCLATHRIN1 and CsUBC1 were the top five appropriate reference genes for qRT-PCR analysis in complex experimental conditions.
authors.affiliation: College of Horticulture, Northwest A&F University, Yangling 712100, Shaanxi, China. xcw75@tricaas.com.
authors.forename: Xinyuan
authors.initials: X
authors.lastname: Hao
authors.affiliation: USDA-Agricultural Research Service, Biosciences Research Lab, Sunflower and Plant Biology Research Unit, 1605 Albrecht Blvd N, Fargo, ND 58102, USA. david.horvath@ars.usda.gov.
authors.forename: David P
authors.initials: DP
authors.lastname: Horvath
authors.affiliation: USDA-Agricultural Research Service, Biosciences Research Lab, Sunflower and Plant Biology Research Unit, 1605 Albrecht Blvd N, Fargo, ND 58102, USA. wun.chao@ars.usda.gov.
authors.forename: Wun S
authors.initials: WS
authors.lastname: Chao
authors.affiliation: College of Horticulture, Northwest A&F University, Yangling 712100, Shaanxi, China. yjyang@tricaas.com.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Tea Research Institute, Chinese Academy of Agricultural Sciences, No. 9 South Meiling Road, Hangzhou 310008, Zhejiang, China. xcw75@tricaas.com.
authors.forename: Xinchao
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Horticulture, Northwest A&F University, Yangling 712100, Shaanxi, China. amery.hao@yahoo.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Xiao
mesh_terms: D028241:Camellia sinensis / Q000235:genetics*; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012015:Reference Standards; D015203:Reproducibility of Results; D013662:Tea / Q000235:genetics*
chemical_list: D013662:Tea