Identification and Evaluation of Reliable Reference Genes for Quantitative Real-Time PCR Analysis in Tea Plant (Camellia sinensis (L.) O. Kuntze)

International Journal of Molecular Sciences

Xinyuan Hao²  David P. Horvath³  Wun S. Chao³  Yajun Yang²  Xinchao Wang¹  Bin Xiao² 

[1] Tea Research Institute, Chinese Academy of Agricultural Sciences, No. 9 South Meiling Road, Hangzhou 310008, Zhejiang, China; E-Mail:;College of Horticulture, Northwest A&F University, Yangling 712100, Shaanxi, China; E-Mail:;USDA-Agricultural Research Service, Biosciences Research Lab, Sunflower and Plant Biology Research Unit, 1605 Albrecht Blvd N, Fargo, ND 58102, USA; E-Mails:

关键词: Camellia sinensis;    reference gene;    qRT-PCR;    tea plant;    gene expression;    normalization;   

DOI  :  10.3390/ijms151222155

来源: mdpi

【 摘 要 】

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 ∆C_T 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.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.