文献类型：期刊文献

英文题名：Validation of candidate reference genes for gene expression normalization in Buchloe dactyloides using quantitative real-time RT-PCR

作者：Li, Wei[1,2] Qian, Yong-qiang[1] Han, Lei[1] Liu, Jun-xiang[1] Li, Zhen-Jian[1] Ju, Guan-sheng[1] Sun, Zhen-yuan[1]

第一作者：Li, Wei

通信作者：Sun, ZY[1]

机构：[1]Chinese Acad Forestry, Res Inst Forestry, State Key Lab Tree Genet & Breeding, Beijing 100091, Peoples R China;[2]Qjngdao Agr Univ, Coll Landscape Architecture & Forestry, Qingdao 266109, Shandong, Peoples R China

年份：2015

卷号：197

起止页码：99-106

外文期刊名：SCIENTIA HORTICULTURAE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000367411400012)】；

基金：The work presented here was supported by the National Natural Science Foundation of China (31100505), grant for National Non-profit Research Institutions of Chinese Academy of Forestry(CAFYBB2012043) and the Beijing Natural Science Foundation (6122031). And We are very grateful to Dr. Xiao-jiao Han and Master Li-yuan Lin (The Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang, People's Republic of China) for good suggestions and help with statistical analysis, and Master Ying-guan Liu (The Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang, People's Republic of China) for suggestions and careful proofreading of the manuscript. We also appreciate the anonymous referees and the editor for their comments and suggestions that helped improve the manuscript.

语种：英文

外文关键词：Buchloe dactyloides; Normalization; Photoperiod; qRT-PCR; Reference gene

摘要：The proper selection of reference genes to normalize the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) results under special experimental conditions is crucial for quantifying gene expression. A systematic reference gene screening was performed using SYBR green qRT-PCR on 10 candidate genes in 27 biological samples of various photoperiodic treatments and different tissues in Buchloe dactyloides. Analysis by 3 different qRT-PCR methods (GeNorm, NormFinder, and BestKeeper) indicated that a combination of beta actin (beta-ACT), dnaj protein gene (DNAJ) and protein phosphatase 2a (PP2A) were the most suitable reference genes across all samples examined. While the expression of the beta-ACT and PP2A were the most stable genes in the long day (LD)-treated plant samples, glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH)and beta-ACT performed more stably in short day (SD) and constant dark (24D)-treated plant samples. beta-ACT and PP2A displayed the most stable expression under constant light (24L) condition. beta-ACT and DNAJ were the optimal reference genes in different tissues. Furthermore, examination of relative expression of a functional gene long hypocotyl 5 (HY5) in LD-treated RNA samples demonstrated additional reference genes contribute to accurate quantitative expression analysis. This study will help the selection of the most stable reference genes for accurate gene expression studies in B. dactyloides. (C) 2015 Published by Elsevier B.V.