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Activation of a DRE-binding transcription factor from Medicago truncatula by deleting a Ser/Thr-rich region

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Abstract

We isolated a DREB homologue gene, MtDREB2A, from Medicago truncatula. Its deduced protein contains an AP2 domain of 59 amino acids. The expression of MtDREB2A was significantly induced in roots by salt and drought treatments. Using megaprimer PCR, we deleted a Ser/Thr-rich coding region between residues 142 and 190, and transformed MtDREB2A to a constitutive form, namely, MtDREB2a. Yeast one-hybrid assay revealed that both MtDREB2A and MtDREB2a specifically bound to the dehydration-responsive element (DRE) and activated the expression of the reporter genes of HIS3 and LacZ. Analysis of transcription activities of the proteins in yeast indicated that MtDREB2a could activate the expression of reporter gene, whereas MtDREB2A could not. Overexpression of MtDREB2a in transgenic M. truncatula resulted in significant dwarfed seedling. These results suggested that MtDREB2A functioned specially in response to salt and drought stresses in M. truncatula and that deletion of the Ser/Thr-rich region between residues 142 and 190 activated the transcriptional activation ability of MtDREB2A.

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Acknowledgements

The authors gratefully acknowledge the laboratory support provided by Professor Xiaofang Luo. We also thank Dr. Jingsong Zhang for guidance on yeast one-hybrid assay. Medicago seeds were sent by Prof. Guo Lanbin from Australia to China. This work is supported by China National “948” Program (Grant No. 2005-4-34, 2007-4-02); National High Technology Program (2006AA10Z182); Special Research Fund for Doctor’s Degree Dissertation in Chinese Universities (20060022012); National Natural Science Foundation of China (30371148).

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Authors and Affiliations

  1. College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing, 100083, People’s Republic of China

    Ji-Ren Chen, Jing-Jing Lü, Tian-Xiang Wang & Hua-Fang Wang

  2. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, People’s Republic of China

    Shou-Yi Chen

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  1. Ji-Ren Chen
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Correspondence to Hua-Fang Wang.

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Editor: F.A. Krens

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Chen, JR., Lü, JJ., Wang, TX. et al. Activation of a DRE-binding transcription factor from Medicago truncatula by deleting a Ser/Thr-rich region. In Vitro Cell.Dev.Biol.-Plant 45, 1–11 (2009). https://doi.org/10.1007/s11627-008-9163-9

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