Effect of a biotic elicitor and nano elicitor on some alkaloids production in Papaver somniferum L.

Document Type : Full Paper


1 Former Graduate Student, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran

2 Professor, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran

3 Associate Professor , University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran

4 Assistant Professor , Department of Pharmacognosy and Pharmaceutics, Medicinal Plants Research Center, Institute of Medicinal Plants, ACECR, Karaj, Iran

5 Expert, Department of Pharmacognosy and Pharmaceutics, Medicinal Plants Research Center, Institute of Medicinal Plants, ACECR, Karaj, Iran


To develop an optimal bioprocess for benzophenanthridin alkaloids production, various metabolite engineering are studied to increase yields, in which one of them is elicitation. Elicitors are biotic or abiotic molecules that have effectively stimulated the production of plant secondary metabolites in cell and organ cultures. Since this plant serves as the only commercial source of morphine and sanguinarine production and chemical synthesis of those alkaloids are not economically feasible, therefor, in this study, after selection of the best media and explant, the effects of two elicitors, nano TiO2 and yeast extract, were studied at different time points (24 and 48h) following treatments on sanguinarine and thebaine production in meristem and root suspension cell cultures. This research was conducted using a randomized complete design with factorial arrangement and three replications. It was observed that these elicitors had the positive effect on accumulation of sanguinarine and thebaine. Elicitation with yeast extract at 48h intervals enhanced the accumulation of sanguinarine (2.2 times) and thebaine (3.4 times) compared to the control. Treatment of nano TiO2 in 24h increased both alkaloids (2.1times) which decreased over the time. This study showed the higher excitability of meristemic cells compared to the root suspension cell culture. Therefore, due to the more provoking and increasing effects on sanguinarine and thebaine content throughout the study, these elicitors and in particular nano one are proved to be promising and suitable candidate for elicitation in both meristem and root cell suspension cultures.


          Karimaneh, Z., Omidi, M., Khialparast, F., & Sabokdast, M. (2009). Study on the effect of plant growth regulators, concentration, medium and explants on the callus induction, regeneration and suspension culture of Papaver somniferum L. M.Sc Thesis. Faculty of agriculture and natural resources, University of Tehran, Iran. (In Farsi).
          Abraham, F., Bhatt, A., Keng, Ch., Indrayanto, G., & Sulaiman, Sh. (2011). Effect of yeast extract and chitosan on shoot proliferation, morphology and antioxidant activity of Curcuma mangga in vitro plantlets. African Journal of Biotechnology, 10 (40),7787-7795.
          Alcantara, J., Bird, D.A., Franceschi, V.R., & Facchini, P.J. (2005). Sanguinarine biosynthesis is associated with the endoplasmic reticulum in cultured opium poppy cells after elicitor treatment. Plant Physiology, 138, 173–183.
          Angelova, Z., Georgiev, S., & Roos, W. (2006). ELICITATION OF PLANTS. Biotechnology & biotechnology, PP, 72-83.
          Archambault, J., Williams, R.D, & Chavarie, C. (1996). Production of sanguinarine by elicited plant cell culture I. Shake flask suspension cultures. Journal of Biotechnology, 46, 95-105.
           Balažová, A., Blanáriková, V., Bilka, F., & Bilková, A. (2008). Effect of Combined Biotic and Abiotic elicitor on the Sanguinarine Formation in Cell Suspension Cultures of Eschscholtzia californica. ACTA Faccultatis Pharmaceutical, 55, 58-63.
          Byun, S.Y., Ryu, Y.W., Kim, C., & Pedersen, H. (1992). Elicitation of sanguinarine production in two-phase cultures of Eschscholtzia californica, Journal of Ferment Bioengineer, 73,380–385.

          Castiglione, R., Giorgetti, M., Geri, L., & Cremonini, C. (2011). The effects of nano-TiO2 on seed germination, development and mitosis of root tip cells of Vicia narbonensis L. and Zea mays L, Journal of Nanoparticle Research, 13,6-7.

           Chaterjee, A., Shukla S., Mishra, P., Rastogi, A., & Singh, S.P. (2010). Prospects of in vitro production of thebaine in opium poppy (Papaver somniferum L.).Industrial Crops and Products,
), 668–670.

          Decker, G., Wanner, G., Zenk, M.H., & Lottspeich, F. (2000). Characterization of proteins in latex of the opium poppy (Papaver somniferum) using two-dimensional gel electrophoresis and microsequencing. Electrophoresis, 21, 3500-3516.
          Desgagné-Penix, I., Khan, M., Schriemer, D., Cram, D., Nowak, J., & Facchini, P. (2010). Integration of deep transcriptome and proteome analyses reveals the components of alkaloid metabolism in opium poppy. Cell cultures, 10, 252-268
          Dumontet, C., & Jordan, MA. (2010). Microtubule-binding agents: a dynamic field of cancer therapeutics. Nature Review Drug Discovery, 9(10),790-803
          Facchini, P., & De Luca, V. (1995). Phloem-specific expression of tyrosine/dopa decarboxylase genes and biosynthesis of isoquinoline alkaloids in opium poppy. Plant Cell, 7, 1811-1821.
          Facchini, P., Penzes, C., Johnson, A., & Bull, D. (1996). Molecular Characterization of Berberine Bridge Enzyme Genes from Opium Poppy. Plant physiology, 112, 1669-1 677.
          Fan, Y., Chao, Liu., Fengqing, G., Mingyu, S., Xiao, W., Lei, Z., Fashui, H. & Ping, Y. (2007). The Improvement of Spinach Growth by Nano-anatase TiO2 Treatment Is Related to Nitrogen Photoreduction. Biological Trace Element Research, 119, 77-88.
          Gottschalk F., Sonderer T., Scholz R.W. & Nowack B. (2009) Modeledenvironmental concentrations of engineered nanomaterials (TiO2, ZnO, Ag, CNT,Fullerenes) for different regions. Environmental Science & Technology, 43, 9216–9222
          Holková, I., Bezáková, L., sek Bilka, F., Bala_zová, A. & Blanáriková, V. (2010). Involvement of lipoxygenase in elicitor-stimulated sanguinarine accumulation in Papaver somniferum suspension cultures. Plant Physiology and Biochemistry, 48, 887-892.
          Hong, F.S., Yang, F., Liu, C., Gao, Q., Wan, Z.G., Gu, F.G., Wu, C., Ma, Z.N., Zhou, J., & Yang, P. (2005). Influences of nano-TiO2 on the chloroplast ageing of spinach under light. Biological trace element research, 104(3), 249–260.
          Huang, F., & Kutchan, T. M. (2000). Distribution of morphinan and benzophenanthridine alkaloid gene transcript accumulation in Papaver somniferum. Phytochemistry, 53, 555-564
          Kavalco, R., Prioli, K., Pinto, S., Prioli, A., & Bertollo, L. (2007). Chromosome polymorphism in Astyanax fasciatus (Teleostei, Characidae), Part 3: Analysis of the RAPD and ISSR molecular markers. Biochemical Systematics and Ecology, 35, 843-851.

          Lei, Z., Mingyu, S., Chao, L., Liang, C., Hao, H., Xiao, W., Xiaoqing, L., Fan, Y., Fengqing, G., & Fashui, H. (2008). Effects of Nanoanatase TiO2 on photosynthesis of spinach chloroplasts under different light illumination photoreduction. Biological Trace Element Research, 119(1), 68-76.

          Liscombe, D.K., & Facchini, P.J. (2008). Evolutionary and cellular webs in benzylisoquinoline alkaloid biosynthesis. Current Opinion in Biotechnology, 19, 173-180.
          Lovkova, M., Buzuk, G.N., & Solkolova, S.M. (2005). Regulatory Role of Elements in the Formation and Accumulation of Alkaloids in Papaver somniferum L. Seedlings. BMC Plant Biology, 10, 252-255.
          Milo, J., Levy, A., & Palevitch D. (2006). Analternative raw- the cultivation breeding ofPapaver bracteatum, The Genus Papaver.Hardwood. Academic Publishers.
          Namdeo. A. G. (2007). Plant Cell Elicitation for Production of Secondary Metabolites Review. Pharmacognosy Reviews, 1, 69-79.
          Radman, R., Saez, T., Bucke, C., & Keshavarz, T. (2003). Elicitation of plants and microbial cell systems. Biotechnology and Applide Biochemistry, 37, 91–102.
          Rush, M. D., Kutchan, T. M, & Coscia, C. J. (1985). Correlation of the appearance of morphinan alkaloids and latificer cells in germinating Papaver bracteatum seedlings. Plant Cell Report, 4, 237-240.  
          Weid, M., Ziegler, J., & Kutchan, T.M. (2004). The roles of latex and the vascular bundle in morphine biosynthesis in the opium poppy, Papaver somniferum. Proceedings of the National Academy of Sciences,101, 13957-13962
          Weiss, D., Baumert, A., Vogel, M., & Roos, W. (2006). Sanguinarine reductase, a key enzyme of benzophenanthridine detoxification. Plant Cell Environment, 29, 291–302.
          Yang, H., Sun, CH., Zhang, Qiao., Zou, J., Liu, G., & Smith, S.C., et al. (2008). Anatase TiO2 single crystals with a large percentage of reactive facets. Nature, 453, 638-641.
          Young, Choa, H., Young Sona, S., Soon Rheea, H., Sung-Yong, H., Yoonb Carolyn, W.T., Parsonsc Jong, L., & Parka M. (2008). Synergistic effects of sequential treatment with methyl jasmonate, salicylic acid and yeast extract on benzophenanthridine alkaloid accumulation and protein expression in Eschscholtzia californica suspension cultures. Journal of Biotechnology, 135, 117–122.
          Zdařilová, A., Malíková, J., Dvořák, Z., Ulrichová, J., & Šimánek, V. )2006(. Quaternary isoquinoline alkaloids sanguinarine and chelerythrine In vitro and in vivo effects. Chemicke Listy, 100, 30-41.
          Zhao, J., & Sakai, K. (2003). Multiple signaling pathways mediate fungal elicitor-induced beta-thujaplicinbiosynthesis in Cupressus lusitanica. cell cultures, 4, 647-656
          Ziegler, J., & Facchini, P. (2008) Alkaloid biosynthesis: metabolism and trafficking. Annual Review of Plant Biology, 59,735-69.