MOLECULAR CLONING, BIOINFORMATION ANALYSIS AND EXPRESSION OF THE STRICTOSIDINE SYNTHASE IN Dendrobium officinale


Abstract

The enzyme strictosidine synthase (STR, EC: 4.3.3.2) plays a key role in the biosynthetic pathway of terpenoid indole alkaloid (TIA). It catalyzes the condensation of the tryptamine and secologanin to form 3α(S)-strictosidine, which is the common precursor of all TIAs. In this paper, a STR gene designated as DoSTR (GenBank: KX068707) was first cloned and characterized from Dendrobium officinale with rapid amplified cDNA ends method (RACE). DoSTR has a length of 1380bp with 1179bp open reading frame encoding 392 amino acids. BlastP analyses showed that its amino acid sequence was classified into Str_synth superfamily. qRT-PCR showed that DoSTR was expressed in all tissues tested, with a significantly higher level in flower and the lowest in stem. Four different treatments with MeJA, SA, ABA and AgNO3, respectively, could induce the DoSTR expression to a different extent. And the effect of MeJA was the most obvious and transcript level of DoSTR induced by MeJA was 20.7 times greater than that of control at 48 hours after treatment. Furthermore, it was found that DoSTR was localized in vacuole through transient expression in tobacco. The characterization and expression of DoSTR can help in further studying the role of DoSTR in the biosynthesis of TIAs in D. officinale. This study may throw light on the alkaloid biosynthesis pathway of D. officinale.


Keywords

Dendrobium officinale; DoSTR; terpenoid indole alkaloid; tissue expression pattern; subcellular localization

Almagro Armenteros, J.J., Tsirigos, K.D., Sønderby, C.K.
(2019). SignalP 5.0 improves signal peptide predictions using deep neural networks. Nat. Biotechnol., 37, 420–423. DOI: 10.1038/s41587-019-0036-z
Artimo, P., Jonnalagedda, M., Arnold, K., Baratin, D., Csardi, G., Castro, E. de, Duvaud, S., Flegel, V., Fortier, A., Gasteiger, E., Grosdidier, A., Hernandez, C., Ioannidis, V., Kuznetsov, D., Liechti, R., Moretti, S., Mostaguir, K., Redaschi, N., Rossier, G., Xenarios, I., Stockinger, H. (2012). ExPASy: SIB bioinformatics resource portal. Nucleic Acids Res., 40(W1), W597–W603. DOI: 10.1093/nar/gks400
Benkert, P., Tosatto, S.C.E., Schomburg, D. (2008). QMEAN: A comprehensive scoring function for model quality assessment. Proteins Struct. Funct. Bioinf., 71, 261–277. DOI: 10.1002/prot.21715
Chen, R., Liao, Z.H., Chen, M., Wang, Q., Yang, C.X., Yang, Y.J. (2008). Molecular cloning and characterization of the Strictosidine synthase Gene from Rauwolfia verticillata. Russ. J. Plant. Physl., 55, 670–675. DOI: 10.1134/s1021443708050117
Chen, X.M., Xiao, S.Y, Guo, S.X. (2006). Comparison of chemical compositions between Dendrobium candidum and Dendrobium nobile. Acta. Acad. Med. Sin., 28(4), 524–529. DOI: 10.1360/aps040120
Cui, L., Ni, X., Ji, Q., Teng, X., Yang, Y., Wu, C., Zekria, D., Zhang, D., Kai, G. (2015). Co-overexpression of geraniol-10-hydroxylase and strictosidine synthase improves anti-cancer drug camptothecin accumulation in Ophiorrhiza pumila. Sci. Rep., 5, 8227. DOI: 10.1038/srep08227
Flores-Sanchez, I.J., Paniagua-Vega, D., Vera-Reyes, I., Cerda-García-Rojas, C.M., Ramos-Valdivia, A.C. (2016). Alkaloid biosynthesis and metabolic profiling responses to jasmonic acid elicitation in Hamelia patens plants by NMR-based metabolomics. Metabolomics, 12, 1–14. DOI: 10.1007/s11306-016-0999-4
Goklany, S., Rizvi, N.F., Loring, R.H., Cram, E.J., Lee-Parsons, C.W.T. (2013). Jasmonate-dependent alkaloid biosynthesis in Catharanthus roseus hairy root cultures is correlated with the relative expression of Orca and Zct transcription factors. Biotechnol. Progr., 29, 1367–1376. DOI: 10.1002/btpr.1801
Guo, X., Li, Y., Li, C., Luo, H., Wang, L., Qian, J., Luo, X., Xiang, L., Song, J., Sun, C., Xu, H., Yao, H., Chen, S. (2013). Analysis of the Dendrobium officinale transcriptome reveals putative alkaloid biosynthetic genes and genetic markers. Gene, 527, 131–138. DOI: 10.1016/j.gene.2013.05.073
He, T., Huang, Y., Yang, L., Liu, T., Gong, W., Wang, X., Sheng, J., Hu, J. (2016). Structural characterization and immunomodulating activity of polysaccharide from Dendrobium officinale. Int. J. Biol. Macromol., 83, 34–41. DOI: 10.1016/j.ijbiomac.2015.11.038
Hicks, M.A., Barber, A.E., Giddings, L., Caldwell, J., O’Connor, S.E., Babbitt, P.C. (2011). The evolution of function in strictosidine synthase-like proteins. Proteins Struct. Funct. Bioinf., 79, 3082–3098. DOI: 10.1002/prot.23135
Huang, K.W., Li, Y.R., Tao, S.C., Wei, G., Huang, Y., Chen, D., Wu, C. (2016). Purification, characterization and biological activity of polysaccharides from Dendrobium officinale. Molecules, 21, 701. DOI: 10.3390/molecules21060701
Irmler, S., Schroder, G., St-Pierre, B., Crouch, N.P., Hotze, M., Schmidt, J., Strack, D., Matern, U., Schroder, J. (2000). Indole alkaloid biosynthesis in Catharanthus roseus: new enzyme activities and identification of cytochrome P450CYP72A1 as secologanin synthase. Plant. J., 24, 797–804. DOI: 10.1111/j.1365-313X.2000.00922.x
Jiao, C.Y., Song, C., Zheng, S.Y., Zhu, Y.P., Jin, Q., Cai, Y.P., Lin, Y. (2018). Metabolic Profiling of Dendrobium officinale in Response to Precursors and Methyl Jasmonate. Int. J. Mol. Sci., 19(3), 728–746. DOI: 10.3390/ijms19030728
http://www.cbs.dtu.dk/services/SignalP/
http://www.expasy.org
http://www.ncbi.nlm.nih.gov/orffinder/
https://www.predictprotein.org/
Kibble, N.A.J., Sohani, M.M., Shirley, N., Byrt, C., Roessner, U., Bacic, A., Schmidt, O., Schultz, C.J. (2009). Phylogenetic analysis and functional characterisation of strictosidine synthase-like genes in Arabidopsis thaliana. Funct. Plant. Biol., 36, 1098–1109. DOI: 10.1071/FP09104
Kutchan, T.M. (1989). Expression of enzymatically active cloned stritosidine synthase from the higher-plant Rauvolfia serpentina in Escherichia coli. Febs. Lett., 257, 127–130. DOI: 10.1016/0014-5793(89)81802-2
LopezMeyer, M., Nessler, C.L. (1997). Tryptophan decarboxylase is encoded by two autonomously regulated genes in Camptotheca acuminata which are differentially expressed during development and stress. Plant. J., 11, 1167–1175. DOI: 10.1046/j.1365-313X.1997.11061167.x
Lorence, A., Nessler, C.L. (2004). Camptothecin, over four decades of surprising findings. Phytochemistry, 65, 2735–2749. DOI: 10.1016/j.phytochem.2004.09.001
Lu, H., McKnight, T.D. (1999). Tissue-specific expression of the beta-subunit of tryptophan synthase in Camptotheca acuminata, an indole alkaloid-producing plant. Plant. Physiol., 120, 43–51. DOI: 10.1104/pp.120.1.43
Lu, Y., Wang, H., Wang, W., Qian, Z., Li, L., Wang, J., Zhou, G., Kai, G. (2009). Molecular characterization and expression analysis of a new cDNA encoding strictosidine synthase from Ophiorrhiza japonica. Mol. Biol. Rep., 36, 1845–1852. DOI: 10.1007/s11033-008-9389-y
Luo, Q., Tang, Z., Zhang, X., Zhong, Y., Yao, S., Wang, L., Lin, C., Luo, X. (2016). Chemical properties and antioxidant activity of a water-soluble polysaccharide from Dendrobium officinale. Int. J. Biol. Macromol., 89, 219–227. DOI: 10.1016/j.ijbiomac.2016.04.067
Ma, X. Panjikar, S., Koeke, J. (2006). The structure of Rauvolfia serpentina strictosidine synthase is a novel six-bladed β-propeller fold in plant proteins. Plant Cell, 18, 907–920. DOI: 10.2307/20076650
Manjunatha, B.L., Singh, H.R., Ravikanth, G., Nataraja, K.N., Shankar, R., Kumar, S., Shaanker, R.U. (2016). Transcriptome analysis of stem wood of Nothapodytes nimmoniana (Graham) Mabb. identifies genes associated with biosynthesis of camptothecin, an anti-carcinogenic molecule. J. Biosci., 41, 119–131. DOI: 10.1007/s12038-016-9591-3
Mariani, V., Kiefer, F., Schmidt, T., Haas, J., Schwede, T. (2011). Assessment of template based protein structure predictions in CASP9. Proteins Struct. Funct. Bioinf., 79, 37–58. DOI: 10.1002/prot.23177
Mcknight, T.D., Bergey, D.R., Burnett, R.J., Nessler, C.L. (1991). Expression of enzymatically active and correctly targeted strictosidine synthase in transgenic tobacco plants. Planta, 185, 148–152. DOI: 10.1007/BF00194055
Mcknight, T.D., Roessner, C.A., Devagupta, R., Scott, A.I., Nessler, C.L. (1990). Nucleotide-sequence of a cDNA-encoding the vacuolar protein strictosidine synthase from Catharanthus roseus. Nucleic. Acids. Res., 18, 4939. DOI: 10.1093/nar/18.16.4939
Ng, T.B., Liu, J., Wong, J.H., Ye, X., Wing Sze, S.C., Tong, Y., Zhang, K.Y. (2012). Review of research on Dendrobium, a prized folk medicine. Appl. Microbiol. Biot., 93, 1795–1803. DOI: 10.1007/s00253-011-3829-7
Pasquali, G., Goddijn, O.J., De, W.A., Verpoorte, R., Schilperoort, R.A. (1992). Coordinated regulation of two indole alkaloid biosynthetic genes from Catharanthus roseus by auxin and elicitors. Plant. Mol. Biol., 18, 1121–1131. DOI: 10.1007/BF00047715
Pauwels, L., Morreel, K., De Witte, E., Lammertyn, F., Van Montagu, M., Boerjan, W., Inzé, D., Goossens, A. (2008). Mapping methyl jasmonate-mediated transcriptional reprogramming of metabolism and cell cycle progression in cultured Arabidopsis cells. PNAS, 105(4), 1380–1385. DOI: 10.1073/pnas.0711203105
Pfaffl, M.W. (2001). A new mathematical model for relative quantification in real-time RT-PCR. Nucleic. Acids. Res., 29, 2002–2009. DOI: 10.1093/nar/29.9.e45
Stevens, L.H., Giroud, C., Pennings, E., Verpoorte, R. (1993). Purification and characterization of strictosidine synthase from a suspension-culture of Cinchona robusta. Phytochemistry, 33, 99–106. DOI: 10.1016/0031-9422(93)85403-E
Venditto, V.J., Simanek, E.E. (2010). Cancer therapies utilizing the camptothecins: a review of the in vivo literature. Mol. Pharm., 7, 307–349. DOI: 10.1021/mp900243b
Wang, C.T., Liu, H.T., Gao, X.S., Zhang, H. (2010). Overexpression of G10H and ORCA3 in the hairy roots of Catharanthus roseus improves catharanthine production. Plant. Cell. Rep., 29, 887–894. DOI: 10.1007/s00299-010-0874-0
Wei, W., Feng, L., Bao, W.R., Ma, D.L., Leung, C.H., Nie, S.P., Han, Q.B. (2016). Structure characterization and immunomodulating effects of polysaccharides isolated from Dendrobium officinale. J. Agric. Food. Chem., 64(4), 881–889. DOI: 10.1021/acs.jafc.5b05180
Wheeler, D.L., Barrett, T., Benson, D.A., Bryant, S.H., Canese, K., Chetvernin, V. (2007). Database resources of the national center for biotechnology information. Nucleic Acids Res., 35(Database), D5–D12. DOI: 10.1093/nar/gkl1031
Wungsintaweekul, J., Choo-malee, J., Charoonratana, T., Keawpradub N. (2012). Methyl jasmonate and yeast extract stimulate mitragynine production in Mitragyna speciosa (Roxb.) Korth. shoot culture. Biotechnol. Lett., 34, 1945–1950. DOI: 10.1007/s10529-012-0968-6
Xu, J., Han, Q., Li, S., Chen, X., Wang, X., Zhao, Z., Chen, H. (2013). Chemistry, bioactivity and quality control of Dendrobium, a commonly used tonic herb in traditional Chinese medicine. Phytochem. Rev., 12, 341–367. DOI: 10.1007/s11101-013-9310-8
Yachdav, G., Kloppmann, E., Kajan, L., Hecht, M., Goldberg, T., Hamp, T., Hönigschmid, P., Schafferhans, A., Roos, M., Bernhofer, M., Richter, L., Ashkenazy, H., Punta, M., Schlessinger, A., Bromberg, Y., Schneider, R., Vriend, G., Sander, Ch., Ben-Tal, N., Rost, B. (2014). PredictProtein – an open resource for online prediction of protein structural and functional features. Nucleic Acids Res., 42(Web Server issue), W337–W343. DOI: 10.1093/nar/gku366
Yamazaki, Y., Sudo, H., Yamazaki, M., Aimi, N., Saito, K. (2003). Camptothecin biosynthetic genes in hairy roots of Ophiorrhiza pumila: cloning, characterization and differential expression in tissues and by stress compounds. Plant. Cell. Physiol., 44, 395–403. DOI: 10.1093/pcp/pcg051
Yan, L., Wang, X., Liu, H., Tian, Y., Lian, J., Yang, R., Hao, S., Wang, X., Yang, S., Li, Q., Qi, S., Kui, L., Okpekum, M., Ma, X., Zhang, J., Ding, Z., Zhang, G., Wang, W., Dong, Y., Sheng, J. (2015). The Genome of Dendrobium officinale Illuminates the Biology of the Important Traditional Chinese Orchid Herb. Mol. Plant., 8, 922–934. DOI:10.1016/j.molp.2014.12.011
Zhang, X., Zhang, S., Gao, B., Qian, Z., Liu, J., Wu, S., Si, J. (2019). Identification and quantitative analysis of phenolic glycosides with antioxidant activity in methanolic extract of Dendrobium catenatum flowers and selection of quality control herb-markers. Food Res. Int., 123, 732–745.
Zhu, N., Han, S., Yang, C., Qu, J., Sun, Z., Liu, W., Zhang, X. (2016). Element-tracing of mineral matters in Dendrobium officinale using ICP-MS and multivariate analysis. SpringerPlus, 5, 1–9. DOI: 10.1186/s40064-016-2618-2
Xu, R.X., Cao, F.X., Peng, J.Q., Si, S.B. (2012). Cloning and analysis of strictosidine synthase in Rauvolfia yunnanensis. J. Cent. South. Univ. For Technol., 32, 128–131. DOI: 10.1007/s11783-011-0280-z
Download

Published : 2020-06-29


Zhu, Y.-F., Fan, H.-H., Li, D.-H., Jin, Q., Zhang, C.-M., Zhu, L.-Q., Song, C., Cai, Y.-P., & Lin, Y. (2020). MOLECULAR CLONING, BIOINFORMATION ANALYSIS AND EXPRESSION OF THE STRICTOSIDINE SYNTHASE IN Dendrobium officinale. Acta Scientiarum Polonorum Hortorum Cultus, 19(3), 111-124. https://doi.org/10.24326/asphc.2020.3.10

Yan-Fang Zhu  yfangok@163.com
Key Laboratory of Resource Plant Biology of Anhui Province, School of Life Sciences, Huaibei Normal University, Huaibei 235000, P.R. China; School of Life Sciences, Anhui Agricultural University, Hefei, 230036, P.R. China  China
Hong-Hong Fan 
School of Life Sciences, Anhui Agricultural University, Hefei, 230036, P.R. China  China
Da-Hui Li 
School of Life Sciences, Anhui Agricultural University, Hefei, 230036, P.R. China  China
Qing Jin 
School of Life Sciences, Anhui Agricultural University, Hefei, 230036, P.R. China  China
Chuan-Ming Zhang 
School of Life Sciences, Anhui Agricultural University, Hefei, 230036, P.R. China  China
Li-Qin Zhu 
School of Life Sciences, Anhui Agricultural University, Hefei, 230036, P.R. China  China
Cheng Song 
School of Life Sciences, Anhui Agricultural University, Hefei, 230036, P.R. China  China
Yong-Ping Cai 
School of Life Sciences, Anhui Agricultural University, Hefei, 230036, P.R. China  China
Yi Lin 
School of Life Sciences, Anhui Agricultural University, Hefei, 230036, P.R. China  China




 

Articles are made available under the CC BY-NC-ND 4.0 (recognition by authorship, non-commercial use, no dependent works).
The author signs a statement on the originality of the work and the contribution of individuals.

Submission of the paper implies that it has not been published previously, that it is not under consideration for publication elsewhere.