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Reference Gene Selection for Real-Time Quantitative Polymerase Chain Reaction of mRNA Transcript Levels in Chinese Cabbage (Brassica rapa L. ssp pekinensis)

文献类型： 外文期刊

作者： Qi, Jiani 1 ; Yu, Shuancang 1 ; Zhang, Fenglan 1 ; Shen, Xiangqun 2 ; Zhao, Xiuyun 1 ; Yu, Yangjun 1 ; Zhang, Deshuang 1 ;

作者机构： 1.BVRC, BAAFS, Beijing 100097, Peoples R China

2.Shenyang Agr Univ, Dept Hort, Shenyang 110161, Peoples R China

关键词： mRNA: messenger RNA;Gene Name;Brassica rapa pekinensis GAPDH gene [Cruciferae]: Brassica rapa pekinensis glyceraldehyde-3-phosphat;Brassica rapa pekinensis Ubc gene [Cruciferae]: Brassica rapa pekinensis ubiquitin gene;Brassica rapa pekinensis EF-1-alpha gen;e [Cruciferae]: Brassica rapa pekinensis elongation-factor-1-alpha gene;Brassica rapa pekinensis Apr gene [Cruciferae]: Brassica rapa pekinensis adenine phosphoribosyl-tran;Brassica rapa pekinensis clathrin gene [Cruciferae];Brassica ra;pa pekinensis cyclophilin gene [Cruciferae];Brassica rapa pekinensis Tub gene [Cruciferae]: Brassica rapa pekinensis tubulin gene;Brassica rapa pekinensis actin gene [Cruciferae];Brassica rapa pekinensis 18s rRNA gene [Cruciferae];Brassica rapa p;ekinensis PSY gene [Cruciferae]: expression;Sequence Data;FJ227935: GenBank;EMBL;DDBJ;amino acid sequence;nucleotide sequence

期刊名称：PLANT MOLECULAR BIOLOGY REPORTER （ 影响因子：1.595； 五年影响因子：2.042 ）

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收录情况： SCI

摘要： The Chinese cabbage is a crop belonging to the family Cruciferae; very few studies have focused on the analysis of gene expression in this economically important crop. In this study, we used real-time quantitative polymerase chain reaction to identify the control genes that are the most stably expressed in a given set of tissues and under conditions of drought stress and downy mildew infection. We characterized the transcript stability of nine candidate reference genes, namely, those encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ubiquitin (Ubc), elongation-factor-1-alpha (EF-1-alpha), adenine phosphoribosyltransferase (Apr), clathrin, cyclophilin (Cyp), tubulin (Tub), Actin, and 18s rRNA, using the geNorm and Normfinder software programs. The results of a geNorm analysis indicated that EF-1-alpha and Apr were the most suitable reference genes among the given set of tissues and that GAPDH and Ubc were the most stable genes under conditions of drought stress and downy mildew infection. Furthermore, the results of an analysis performed using the Normfinder software program indicated EF-1-alpha to be the best normalization factor in the given set of tissues and under conditions of downy mildew infection and GAPDH to be the best factor under drought stress conditions. The results of the geNorm analysis also helped determine the minimum number of genes required to calculate a reliable normalization factor. A study on the variability of expression of PSY expression showed that the relative quantification of this gene varied depending on the internal control and the number of internal controls used, thus highlighting the importance of the choice of internal controls in such experiments.

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