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Characterization and expression profiling of 4-hydroxyphenylpyruvate dioxygenase gene (Smhppd) from Salvia miltiorrhiza hairy root cultures

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Abstract

A novel 4-hydroxyphenylpyruvate dioxygenase gene (designated as Smhppd) was cloned from hairy roots of Salvia miltiorrhiza Bung. The full-length cDNA of Smhppd was 1,736 bp long with an ORF (open reading frame) that putatively encoded a polypeptide of 481 amino acids, with a predicted molecular mass of 52.54 kDa. The deduced amino acid sequence of the Smhppd gene shared high homology with other known HPPDs. Analysis of Smhppd genomic DNA revealed that it contained two exons and one intron. The analysis of Smhppd promoter region was also presented. Southern-blot analysis revealed that the Smhppd was a low-copy gene in S. miltiorrhiza. Real-time quantitative PCR analysis indicated that Smhppd was constitutively expressed in roots, stems and leaves of S. miltiorrhiza, with the high expression in roots. In addition, Smhppd expreession level under different stress condition was also analyzed during the hairy root culture period, including signaling components for plant defence responses, such as methyl jasmonate and salicylic acid, as well as an abiotic elicitor, Ag+ and a biotic elicitor, yeast extract. This study will enable us to further understand the role Smhppd plays in the synthesis of active pharmaceutical compounds in S. miltiorrhiza at molecular level.

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Acknowledgments

This research was financially supported by National Natural Science Foundation of China (20572130, 30600807), Shanghai Natural Science Foundation (04ZR14051), Shanghai Science and Technology Committee, and Shanghai Rising-Star Program (06QB14010).

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Authors and Affiliations

  1. Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai, 200433, China

    Ying Xiao, Ying Liu & Lei Zhang

  2. Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China

    Ying Xiao, Peng Di, Junfeng Chen, Ying Liu & Wansheng Chen

  3. Modern Research Center for Traditional Chinese Medicine, Second Military Medical University, Shanghai, 200433, China

    Junfeng Chen & Wansheng Chen

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  1. Ying Xiao
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  2. Peng Di
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  3. Junfeng Chen
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Correspondence to Wansheng Chen or Lei Zhang.

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Xiao, Y., Di, P., Chen, J. et al. Characterization and expression profiling of 4-hydroxyphenylpyruvate dioxygenase gene (Smhppd) from Salvia miltiorrhiza hairy root cultures. Mol Biol Rep 36, 2019–2029 (2009). https://doi.org/10.1007/s11033-008-9413-2

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