Selection of reliable reference genes for gene expression studies in Rhododendron micranthum Turcz

文献类型： 外文期刊

第一作者： Yi, Shanjun

作者： Yi, Shanjun;Qian, Yongqiang;Han, Lei;Sun, Zhenyuan;Liu, Junxiang;Ju, Guansheng;Fan, Chengming

作者机构：

关键词： Quantitative real-time PCR;Reference gene;geNorm;NormFinder;Rhododendron micranthum Turcz

期刊名称：SCIENTIA HORTICULTURAE （ 影响因子：3.463； 五年影响因子：3.672 ）

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收录情况： SCI

摘要： Selection of reliable reference genes is a prerequisite for accurate normalization of gene expression in quantitative real-time polymerase chain reaction (qRT-PCR). The present study is the first report of a systematic evaluation of potential reference genes in Rhododendron micranthum Turcz. The expression stabilities of 6 candidate reference genes (185, ACT, CYP, UBQ TUA, and EF1 alpha) in different tissues and the same tissue at different developmental stages were assessed in Rh. micranthum Turcz, using two analysis software packages, geNorm and NormFinder. The results indicated that EF1 alpha and UBQ were the most stable reference genes for different tissues (leaf, root, stem and flower) at the same developmental stage, whereas 18S should be avoided. During leaf development. EF1 alpha and 185 can be used as the most reliable reference genes, but CYP was the most variable one. During flower development, the most stable genes were CYP and EF1 alpha, while 18S was the least stable one. In order to demonstrate the importance of using suitable reference genes for obtaining an accurate and reliable results in gene expression studies, the expression pattern of an independent target gene, RhMDH gene was analyzed in parallel, the RhMDH showed an expression difference of 58-fold in stem when normalized with the most stable gene (EF1 alpha)or the least stable gene (185) act as reference, respectively. Our results support that the selection of suitable reference genes is necessary to achieve accurate results in qRT-PCR. (C) 2012 Elsevier B.V

分类号： S6

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