番茄在非生物胁迫下实时定量RT-PCR中内参基因的筛选

Screening the Reference Genes for the Studies of Quantitative Real-time RT-PCR in Tomato under Abiotic Stress

近年来实时荧光定量PCR已经成为研究基因表达的标准方法,稳定的内参基因可使反应标准化进行,并提高该方法的敏感度和重复性。许多研究表明不同组织、细胞和不同条件下内参基因的表达存在很大差异,因此在研究基因表达分析时,需要以内参基因对目标基因的表达量进行校准,由此来获得更为准确的结果。本研究以番茄经过果糖、蔗糖、葡萄糖、高温和低温处理的材料为研究对象,利用实时荧光定量PCR方法,对Actin、CAC、TIP41、Expressed、SAND 5个内参基因m RNA水平的表达量进行分析。经ge Norm和Norm Finder软件分析5个内参基因的表达稳定性,以期为番茄果实基因表达调控相关的研究提供内参基因。研究结果为番茄植株在非生物胁迫下的实时定量RT-PCR分析中内参基因的选择提供了理论与实验依据。

I n recent years, real-time PCR has become the standard method for gene expression studies, stable reference gene can standardize the reaction of RT PCR to further improve the sensitivity and reproducibility of the method. However, many studies have shown that there was a big difference between different tissues, cells and conditions for internal gene expression. Therefore, in order to obtain more accurate results in the study of gene expression analysis, reference gene is in need for calibrating the standard of target gene. In this study, five housekeeping genes such as Actin, CAC, TIP41, Expressed and SAND were employed to be assessed by real-time RT-PCR by using tomato as research material under the treatment of fructose, sucrose, glucose, high and low temperature treatments to analyze the expression stability of five reference genes based on geNorm and NormFinder program. The results of this research may provide theoretical and experimental evidences for the selection of reference genes in the studies of tomato fruit gene regulation under the abiotic stresses.