Submitted to: The Plant Genome
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/30/2008
Publication Date: 7/16/2008
Citation: Libault, M., Thibivilliers, S., Radwan, O., Clough, S.J., Stacey, G. 2008. Identification of four soybean reference genes for gene expression normalization. The Plant Genome. 1:44-54. Interpretive Summary: The activation and repression of gene expression controls how an organism responds to a given treatment. When working with a small set of genes, one needs to have a control gene that is not affected by the treatment(s) of interest to standardize the expression experiment. Currently, there are only a few genes that have been proven to be good control genes that are constitutively expressed across many treatments and tissue type. We identified four very good constitutive control genes that soybean researchers can use in their expression studies.
Technical Abstract: Gene expression analysis requires the use of reference genes stably expressed independently of specific tissues or environmental conditions. Housekeeping genes (e.g., actin, tubulin, ribosomal, polyubiquitin and elongation factor 1-alpha) are commonly used as reference genes with the assumption that they are stably expressed. However, in many cases, this assumption has been shown to be incorrect. To avoid these issues and to provide more constitutive control options that one may consider for their specific study, we surveyed a set of genes that showed little variation in a nodulation study across a series of soybean microarray experiments and identified 18 putative soybean reference genes that showed consistent expression levels across experiments. The expression of these 18 genes was further analyzed by quantitative RT-PCR. This analysis confirmed the stable expression of four soybean genes (i.e., annotated as: pectinesterase like protein, F-box protein family, putative zinc protease, CDPK-related protein kinase). These genes should be useful for normalization of gene expression studies in soybean, an important crop plant.