title: Reference Gene Selection for qPCR Normalization of Kosteletzkya virginica under Salt Stress.
abstract: Kosteletzkya virginica (L.) is a newly introduced perennial halophytic plant. Presently, reverse transcription quantitative real-time PCR (qPCR) is regarded as the best choice for analyzing gene expression and its accuracy mainly depends on the reference genes which are used for gene expression normalization. In this study, we employed qPCR to select the most stable reference gene in K. virginica which showed stable expression profiles under our experimental conditions. The candidate reference genes were 18S ribosomal RNA (18SrRNA), β-actin (ACT), α-tubulin (TUA), and elongation factor (EF). We tracked the gene expression profiles of the candidate genes and analyzed their stabilities through BestKeeper, geNorm, and NormFinder software programs. The results of the three programs were identical and 18SrRNA was assessed to be the most stable reference gene in this study. However, TUA was identified to be the most unstable. Our study proved again that the traditional reference genes indeed displayed a certain degree of variations under given experimental conditions. Importantly, our research also provides guidance for selecting most suitable reference genes and lays the foundation for further studies in K. virginica.
authors.affiliation: Key Laboratory of Coastal Biology & Bioresources Utilization, Yantai Institute of Coastal Zone Research (YIC), Chinese Academy of Sciences (CAS), Yantai 264003, China ; University of Chinese Academy of Sciences, Beijing 100039, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Tang
authors.affiliation: Key Laboratory of Coastal Biology & Bioresources Utilization, Yantai Institute of Coastal Zone Research (YIC), Chinese Academy of Sciences (CAS), Yantai 264003, China ; University of Chinese Academy of Sciences, Beijing 100039, China ; Yantai Academy of China Agricultural University, Yantai 264670, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Shandong Agricultural University, Taian 271018, China.
authors.forename: Chuyang
authors.initials: C
authors.lastname: Shao
authors.affiliation: Key Laboratory of Coastal Biology & Bioresources Utilization, Yantai Institute of Coastal Zone Research (YIC), Chinese Academy of Sciences (CAS), Yantai 264003, China ; Institute of Agro-Biotechnology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
authors.forename: Hongbo
authors.initials: H
authors.lastname: Shao
mesh_terms: D000199:Actins / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D019660:Malvaceae / Q000235:genetics*; D010445:Peptide Elongation Factors / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012015:Reference Standards; D012492:Salts / Q000633:toxicity; D012965:Sodium Chloride / Q000633:toxicity; D013312:Stress, Physiological / Q000235:genetics*
chemical_list: D000199:Actins; D010445:Peptide Elongation Factors; D012337:RNA, Ribosomal, 18S; D012492:Salts; D012965:Sodium Chloride