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Reference Gene Selection in the Desert Plant Eremosparton songoricum
Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, Urumqi 830011, China
Graduate University of Chinese Academy of Sciences, Beijing 100049, China
Department of Plant Biology, Southern Illinois University, Carbondale, IL 62901, USA
* Author to whom correspondence should be addressed.
Received: 17 February 2012; in revised form: 3 May 2012 / Accepted: 30 May 2012 / Published: 7 June 2012
Abstract: Eremosparton songoricum (Litv.) Vass. (E. songoricum) is a rare and extremely drought-tolerant desert plant that holds promise as a model organism for the identification of genes associated with water deficit stress. Here, we cloned and evaluated the expression of eight candidate reference genes using quantitative real-time reverse transcriptase polymerase chain reactions. The expression of these candidate reference genes was analyzed in a diverse set of 20 samples including various E. songoricum plant tissues exposed to multiple environmental stresses. GeNorm analysis indicated that expression stability varied between the reference genes in the different experimental conditions, but the two most stable reference genes were sufficient for normalization in most conditions. EsEF and Esα-TUB were sufficient for various stress conditions, EsEF and EsACT were suitable for samples of differing germination stages, and EsGAPDHand EsUBQ were most stable across multiple adult tissue samples. The Es18S gene was unsuitable as a reference gene in our analysis. In addition, the expression level of the drought-stress related transcription factor EsDREB2 verified the utility of E. songoricum reference genes and indicated that no single gene was adequate for normalization on its own. This is the first systematic report on the selection of reference genes in E. songoricum, and these data will facilitate future work on gene expression in this species.
Keywords: Eremosparton songoricum; quantitative real-time PCR; reference genes; geNorm; validation
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Li, X.-S.; Yang, H.-L.; Zhang, D.-Y.; Zhang, Y.-M.; Wood, A.J. Reference Gene Selection in the Desert Plant Eremosparton songoricum. Int. J. Mol. Sci. 2012, 13, 6944-6963.
Li X-S, Yang H-L, Zhang D-Y, Zhang Y-M, Wood AJ. Reference Gene Selection in the Desert Plant Eremosparton songoricum. International Journal of Molecular Sciences. 2012; 13(6):6944-6963.
Li, Xiao-Shuang; Yang, Hong-Lan; Zhang, Dao-Yuan; Zhang, Yuan-Ming; Wood, Andrew J. 2012. "Reference Gene Selection in the Desert Plant Eremosparton songoricum." Int. J. Mol. Sci. 13, no. 6: 6944-6963.