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Analysis of the spatial and temporal expression pattern directed by the Populus tomentosa 4-coumarate:CoA ligase Pto4CL2 promoter in transgenic tobacco

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Abstract

4-Coumarate:CoA ligase (4CL) is a key enzyme in the phenylpropanoid synthesis pathway. The Pto4CL2 promoter was cloned from Populus tomentosa Carr. and fused to the reporter gene encoding β-glucuronidase (GUS); the complex expression patterns directed by the Pto4CL2 promoter were then characterized in Nicotiana tabacum Xanthi by histochemical assays. The promoter 5′-deletion and histochemical assay conducted on transformants indicated that the −317 to −292 nt region supports Pto4CL2 expression in the epidermis and petals and the deletion of the −266 to −252 nt region resulted in the loss of tissue specificity and a dramatic reduction in GUS activity. Furthermore, electrophoretic mobility shift assays testified that an adenine and cytosine-rich element (−264 to −255 nt) and an abscisic acid-responsive element (−242 to −235 nt) in the Pto4CL2 promoter would have functions for the complex expression profiling and efficient basal expression, respectively. These results further clarify the mode of the regulatory expression of class II 4CL promoters in higher plants.

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Acknowledgments

This work was supported by the Fundamental Research Funds for the Central Universities (TD2012-02 and HJ2010-8), the National Natural Science Foundation of China (31170574 and30671697) and the National Basic Research Program of China (2009CB119104).

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Authors and Affiliations

  1. College of Life Sciences and Biotechnology, Beijing Forestry University, Qinghua East Road No. 35, Haidian District, Beijing, 100083, China

    Xiang Pan, Huanhuan Li, Hongyi Wei, Wankai Su, Xiangning Jiang & Hai Lu

  2. National Engineering Laboratory for Tree Breeding, Beijing, 100083, China

    Xiangning Jiang & Hai Lu

  3. Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing, 100083, China

    Hai Lu

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  1. Xiang Pan
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  2. Huanhuan Li
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Correspondence to Hai Lu.

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Pan, X., Li, H., Wei, H. et al. Analysis of the spatial and temporal expression pattern directed by the Populus tomentosa 4-coumarate:CoA ligase Pto4CL2 promoter in transgenic tobacco. Mol Biol Rep 40, 2309–2317 (2013). https://doi.org/10.1007/s11033-012-2312-6

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  • DOI: https://doi.org/10.1007/s11033-012-2312-6

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