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Molecular cloning and characterization of the Strictosidine synthase Gene from Rauwolfia verticillata

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Abstract

The strictosidine synthase (STR, EC 4.3.3.2) catalyzes the condensation of tryptamine and secologanin to form strictosidine, which is the universal precursor for a wide range of pharmaceutical terpenoid indole alkaloids (TIAs). The full-length cDNA encoding STR was cloned and characterized from Rauwolfia verticillata a Chinese native plant producing TIAs, such as reserpine and ajmalicine. The new cDNA was designed as RvSTR and submitted to GenBank to get an accession number DQ017054. The full-length cDNA of RvSTR was 1211 bp containing a 1035-bp open reading frame encoding a deduced 344-amino-acid polypeptide with a calculated mol wt of 38.2 kD and an isoelectric point of 5.19. Comparative and bioinformatic analysis revealed that RvSTR showed a higher similarity to STRs from Apocynaceae species, including Catharanthus roseus and Ophiorrhiza pumila, but a relatively lower similarity to other plant STRs. The unique essential catalytic residue Glu-309 was conserved in all alignment plant species. The phylogenetic analysis revealed that STRs were divided into two groups, including plant and bacterial enzymes. The tissue expression pattern analysis indicated that RvSTR expression could be detected in all tested organs of R. verticillata, including roots, stems, leaves, fruits, and flowers. The lowest transcription level was observed in flowers and the highest was found in fruits; subsequently, the order of transcription level decrease was stems > roots > leaves. The cloning and characterization of RvSTR give a new STR sequence involved in TIA biosynthesis of R. verticillata, and provide a candidate gene for metabolic engineering of the TIA pathway in R. verticillata.

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Abbreviations

ORF:

open reading frame

RACE:

rapid amplification of cDNA ends

STR:

strictosidine synthase

TIA:

terpenoid indole alkaloid.

References

  1. Memelink, J., Verpoorte, R., and Kijne, J.W., Organization of Jasmonate Responsive Gene Expression in Alkaloid Metabolism, Trends Plant Sci., 2001, vol. 6, pp. 212–219.

    Article  PubMed  CAS  Google Scholar 

  2. Nguen, K.K. and Nikolaeva, L.A., Alkaloids of Rauwolfia Species Growing in Vietnam, Chem. Nat. Compd., 1992, vol. 27, pp. 813–816.

    Google Scholar 

  3. Li, H.Y. and Ting, K.S., Haemodynamic Studies on the Total Alkaloids of Rauwolfia verticillata, Sci. Sinica, 1962, vol. 11, pp. 791–804.

    PubMed  CAS  Google Scholar 

  4. Stöckigt, J. and Ruppert, M., Strictosidine: The Biosynthetic Key to Monoterpenoid Indole Alkaloids, Comprehensive Natural Products Chemistry: Amino Acids, Peptides, Porphyrins and Alkaloids, Barton, D.H.R., Ed., Amsterdam: Elsevier, 1999, pp. 109–138.

    Google Scholar 

  5. Kutchan, T.M., Strictosidine: From Alkaloid to Enzyme to Gene, Phytochemistry, 1993, vol. 32, pp. 493–506.

    Article  PubMed  CAS  Google Scholar 

  6. Su, Y.F. and Guo, D., Recent Progress in Biosynthesis of Rauwolfia Alkaloids, J. Chin. Pharm. Sci., 2000, vol. 9, pp. 50–54.

    CAS  Google Scholar 

  7. Silvestrine, A., Pasqua, G., Botta, B., Monacelli, B., van der Heijden, R., and Verpoorte, R., Effects of Alkaloid Precursor Feeding on a Camptotheca acuminata Cell Line, Plant Physiol. Biochem., 2002, vol. 40, pp. 749–753.

    Article  Google Scholar 

  8. Geerlings, A., Hallard, D., van der Heijden, R., and Verpoorte, R., Alkaloid Production by a Cinchona officinalis ‘Ledgeriana’ Hairy Root Culture Containing Constitutive Expression Constructs of Tryptophan Decarboxylase and Strictosidine Synthase cDNAs from Catharanthus roseus, Plant Cell Rep., 1999, vol. 19, pp. 191–196.

    Article  CAS  Google Scholar 

  9. Liao, Z.H., Chen, M., Guo, L., Gong, Y.F., Tang, F., Sun, X.F., and Tang, K.X., Rapid Isolation of Good-Quality Total RNA from Taxus and Ginkgo, Prep. Biochem. Biotechnol., 2004, vol. 34, pp. 209–214.

    Article  PubMed  CAS  Google Scholar 

  10. Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W., and Lipman, D.J., Gapped BLAST and PSI-BLAST: A New Generation of Protein Database Search Programs, Nucleic Acids Res., 1997, vol. 25, pp. 3389–3402.

    Article  PubMed  CAS  Google Scholar 

  11. Thompson, J.D., Higgins, D.G., and Gibson, T.J., CLUSTAL W: Improving the Sensitivity of Progressive Multiple Sequence Alignment through Sequence Weighting, Position-Specific Gap Penalties and Weight Matrix Choice, Nucleic Acids Res., 1994, vol. 22, pp. 4673–4680.

    Article  PubMed  CAS  Google Scholar 

  12. Kumar, S., Tamura, K., and Nei, M., MEGA3: Integrated Software for Molecular Evolutionary Genetics Analysis and Sequence Alignment, Brie BioInform., 2004, vol. 5, pp. 150–163.

    Article  CAS  Google Scholar 

  13. Saitou, N. and Nei, M., The Neighbor-Jointing Method: A New Method for Reconstructing Phylogenetic Trees, Mol. Biol., 1987, vol. 4, pp. 406–425.

    CAS  Google Scholar 

  14. Pasquali, G., Goddijn, O.J., de Waal, A., Verpoorte, R., Schilperoort, R.A., Hoge, J.H., and Memelink, J., Coordinated Regulation of Two Indole Alkaloid Biosynthetic Genes from Catharanthus roseus by Auxin and Elicitors, Plant Mol. Biol., 1992, vol. 18, pp. 1121–1131.

    Article  PubMed  CAS  Google Scholar 

  15. Yamazaki, Y., Sudo, H., Yamazaki, M., Aimi, N., and Saito, K., Camptothecin Biosynthetic Genes in Hairy Roots of Ophiorrhiza pumila: Cloning, Characterization and Differential Expression in Tissues and by Stress Compounds, Plant Cell Physiol., 2003, vol. 44, pp. 395–403.

    Article  PubMed  CAS  Google Scholar 

  16. Ma, X., Panjikar, S., Koepke, J., Loris, E., and Stǒckigtä, J., The Structure of Rauwolfia serpentina Strictosidine Synthase Is a Novel Six-Bladed b-Propeller Fold in Plant Proteins, Plant Cell, 2006, vol. 18, pp. 907–920.

    Article  PubMed  CAS  Google Scholar 

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Correspondence to M. Chen.

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Original Russian Text © R. Chen, Z.H. Liao, M. Chen, Q. Wang, C.X. Yang, Y.J. Yang, 2008, published in Fiziologiya Rastenii, 2008, Vol. 55, No. 5, pp. 746–751.

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Chen, R., Liao, Z.H., Chen, M. et al. Molecular cloning and characterization of the Strictosidine synthase Gene from Rauwolfia verticillata . Russ J Plant Physiol 55, 670–675 (2008). https://doi.org/10.1134/S1021443708050117

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