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Effect of Yeast Extract on h6h Gene Expression and Tropane Alkaloids Production in Atropa belladonna L. Hairy Roots

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Abstract

Plant alkaloids, are one of the largest groups of pharmaceutical metabolites. Scopolamine and atropine are most common tropane alkaloids in Atropa belladonna L. with great application in pharmaceutical industry. Phytochemical biosynthetic pathways induction via various elicitors is an effective strategy for increasing the production of valuable substances. Therefore, in this research effect of yeast extract were studied on expression of h6h gene, antioxidant enzymes activity and production of tropane alkaloid in A. belladonna hairy roots. Hairy roots of A. belladonna were obtained from cotyledonary leaf explants by A7 strain of Agrobacterium rhizogenes. Polymerase chain reaction (PCR) analysis with specific primers confirmed the insertion of rolB in putative transgenic hairy roots genome. Different concentrations of yeast extract (YE) (0, 0.5, 1 and 1.5 mg/L) at different exposure time (24 and 48 h) were used as elicitor. Scopolamine and atropine content and activity of antioxidant enzymes such as guaiacol peroxidase, ascorbate peroxidase and catalase increased and expression of h6h (hyoscyamine 6 beta-hydroxylase) gene in elicited hairy roots were investigated. High concentration of YE decreased the growth of hairy roots but increased the atropine content compared to the control. According to the results, the highest content of scopolamine (9.21%) and atropine (43.39%) which were 9 and 5 fold than control, were obtained by 1 and 1.5 mg/L YE at 48 h exposure time, respectively. The results demonstrated that YE can be used as effective elicitor to increase tropane alkaloids production in hairy root cultures of A. belladonna.

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REFERENCES

  1. Yang, C.X., Chen, M., Zeng, J., Zhang, L., and Liu, X.Q., Improvement of tropane alkaloids production in hairy root cultures of Atropa belladonna by overexpressing pmt and h6h genes. Plant Omics, 2011, vol. 4, p. 29.

    CAS  Google Scholar 

  2. Miraldi, E., Masti, A., Ferri, S., and Comparini, I., Distribution of hyoscyamine and scopolamine in Datura stramonium, Fitoterapia, 2001, vol. 72, p. 644.

    Article  CAS  Google Scholar 

  3. Palazon, J., Moyano, E., Bonfill, M., Cusido, R.M., and Pinol, M.T., Tropane alkaloids in plants and genetic engineering of their biosynthesis, in Floriculture, Ornament-al and Plant Biotechnology: Advances and Topical Issues, Teixeira da Silva, J.A., Ed., London: Global Science Books, 2006, p. 209.

    Google Scholar 

  4. Sharafi, A., Hashemi, S.H., Mousavi, A., Azadi, P., and Hosseini, K.B., Metabolic engineering of morphinan alkaloids by over expression of codeinone reductase in transgenic hairy root of Papaver bracteatum, Biotechnol. Lett., 2013, vol. 35, p. 445.

    Article  CAS  Google Scholar 

  5. Arroo, R., Woolley, J., and Oksman-Caldentey, K.M., Henbane, belladonna, datura and duboisia, in Transgenic Crops VI, Biotechnology in Agriculture and Forestry Series vol. 61, Pua, E.C. and Davey, M.R., Eds., Berlin: Springer-Verlag, 2007, p. 189.

    Google Scholar 

  6. Srivastava, S. and Srivastava, A.K., Production of the biopesticide azadirachtin by hairy root cultivation of Azadirachta indica in liquid-phase bioreactors, App. Biochem. Biotechnol., 2013, vol. 171, p. 1351.

    Article  CAS  Google Scholar 

  7. Pirian, K. and Piri, K., Influence of yeast extract as a biotic elicitor on noradrenaline production in hairy root culture of Portulaca oleracea L., Int. J. Agric. Plant Prod., 2013, vol. 4, p. 2960.

    Google Scholar 

  8. Moharrami, F., Hosseini, B., and Farjaminezhad, M., Enhanced production of hyoscyamine and scopolamine from genetically transformed root culture of Hyoscyamus reticulates L. elicited by iron oxide nanoparticles, In Vitro Cell. Dev. Biol.: Plant, 2017, vol. 53, p. 104.

    Article  CAS  Google Scholar 

  9. Shakeran, Z., Keyhanfar, M., Asghari, G., and Ghanadian, M., Improvement of atropine production by different biotic and abiotic elicitors in hairy root cultures of Datura metel, Turk. J. Biol., 2015, vol. 39, p. 111.

    Article  CAS  Google Scholar 

  10. Shinde, A.M., Malpathak, N., and Fulzele, D.P., Enhanced production of phytoestrogenic isoflavones from hairy root cultures of Psoralea corylifolia L. using elicitation and precursor feeding, Biotechnol. Bioprocess Eng., 2009, vol. 14, p. 288.

    Article  CAS  Google Scholar 

  11. Udomsuk, L., Jarukamjorn, K., Tanaka, H., and Putalun, W., Improved isoflavonoid production in Pueraria candollei hairy root cultures using elicitation, Biotechnol. Lett., 2011, vol. 33, p. 369.

    Article  CAS  Google Scholar 

  12. Singleton, V.L., Orthofer, R., and Lamuela-Raventos, R.M., Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin–Ciocalteu reagent, Methods Enzymol., 1999, vol. 299, p. 152.

    Article  CAS  Google Scholar 

  13. MacAdam, W., Nelson, C.J., and Sharp, R., Peroxidase activity in the leaf elongation zone of Tall fescue (I), Plant Physiol., 1992, vol. 99, p. 872.

    Article  CAS  Google Scholar 

  14. Nakano, Y. and Asada, K., Purification of ascorbate peroxidase in Spinach chloroplasts: its inactivation in ascorbate-depleted medium and reactivation by monodehydroascorbate radical, Plant Cell Physiol., 1987, vol. 28, p. 131.

    CAS  Google Scholar 

  15. Aebi, H., Catalase in vitro, Methods Enzymol., 1984, vol. 105, p. 121.

    Article  CAS  Google Scholar 

  16. Kamada, H., Okamura, N., Satake, M., Harada, H., and Shimomur, K., Alkaloid production by hairy root cultures in Atropa belladonna, Plant Cell Rep., 1986, vol. 5, p. 239.

    Article  CAS  Google Scholar 

  17. Nourozi, E., Hosseini, B., and Hassani, A., A reliable and efficient protocol for induction of hairy roots in Agastache foeniculum, Biologia, 2014, vol. 69, p. 870.

    Article  CAS  Google Scholar 

  18. Samadi, A., Carapetian, J., Heidary, R., Gafari, M., and Hssanzadeh, A., Hairy root induction in Linum mucronatum ssp. mucronatum an anti-tumor lignans production plant, Not. Bot. Horti Agrobot. Cluj-Napoca, 2012, vol. 40, p. 125.

    Article  CAS  Google Scholar 

  19. Li,M., Liao, X., Zhang, D., Du, G., and Chen, J., Yeast extract promotes cell growth and induces production of polyvinyl alcohol-degrading enzymes, Enzyme Res., 2011, vol. 2011, p. 1.

    Google Scholar 

  20. Khan, S., Irfan, Q.M., Kamaluddin, A.T., and Abdin, M.Z., Protocol for isolation of genomic DNA from dry and fresh roots of medicinal plants suitable for RAPD and restriction digestion, Afr. J. Biotechnol., 2007, vol. 6, p. 175.

    CAS  Google Scholar 

  21. Xing, B., Yang, D., Guo, W., Liang, Z. and Yan, X., Ag+ ions as a more effective elicitor for production of tanshinones than phenolic acids in Salvia miltiorrhiza hairy roots, Molecules, 2015, vol. 20, p. 309.

    Article  Google Scholar 

  22. Chamnongpol, S., Willekens, H., Moede, W., Langebartels, C., and Sandermann, H.J.R., Defense activation and enhanced pathogen tolerance induced by H2O2 in transgenic tobacco, Proc. Natl. Acad. Sci. U.S.A., 1998, vol. 95, p. 5818.

    Article  CAS  Google Scholar 

  23. Ghorbanpour, M., Hatami, M., and Hatami, M., Activating antioxidant enzymes, hyoscyamine and scopolamine biosynthesis of Hyoscyamus niger L. plants with nano-sized titanium dioxide and bulk application, Acta Agric. Slov., 2015, vol. 105, p. 23.

    Article  Google Scholar 

  24. dos Santos Soares, A.M., de Souza, T.F., Jacinto, T., and Machado, O.L.T., Effect of methyl jasmonate on antioxidative enzyme activities and on the contents of ROS and H2O2 in Ricinus communis leaves, Braz. J. Plant Physiol., 2010, vol. 22, p. 151.

    Article  Google Scholar 

  25. Thiruvengadam, M., Praveen, N., Maria, J.K.M., Yang, Y.S., and Kim, S.H., Establishment of Momordica charantia hairy root cultures for the production of phenolic compounds and determination of their biological activities, Plant Cell Tissue Organ Cult., 2014, vol.118, p. 545.

    Article  CAS  Google Scholar 

  26. Gundlach, H., Müller, M.J., Kutchan, T.M., and Zenk, M.H., Jasmonic acid is a signal transducer in elicitor-induced plant cell cultures, Proc. Natl. Acad. Sci. U.S.A., 1992, vol. 89, p. 2389.

    Article  CAS  Google Scholar 

  27. Zabetakis, I., Edwards, R., and O’Hagan, D., Elicitation of tropane alkaloid biosynthesis in transformed root cultures of Datura stramonium, Phytochemistry, 1999, vol. 50, p. 53.

    Article  CAS  Google Scholar 

  28. Elshorbagy, M., Ibrahim, S.M., Abo El-Seoud, K.A., and Abd El-Maksoud, A.I., Tropane alkaloids production from callus culture of Atropa belladonna as affected by elicitors and precursor feeding, Int. J. Pharm. Sci. Rev. Res., 2018, vol. 9, p. 116.

    Article  CAS  Google Scholar 

  29. Kheradmand, P.M., Shahriari Ahmadi, F., and Moshtaghi, N., The effect of methyl jasmonate elicitor on tropan alkaloid production in hairy roots of Atropa belladonna, Agric. Biotechnol., 2017, vol. 9, p. 62.

    Google Scholar 

  30. Li, J., Brader, G., and Palva, E.T., The WRKY70 transcription factor: a node of convergence for jasmonate-mediated and salicylate-mediated signals in plant defense, Plant Cell, 2004, vol. 16, p. 319.

    Article  CAS  Google Scholar 

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ACKNOWLEDGMENTS

The authors thank for the West Azarbayjan Branch of Academic Center for Education, Culture and Research (ACECR) for support this study.

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Authors and Affiliations

  1. Academic Center for Education, Culture and Research (ACECR), West Azarbayjan Branch, Urmia, Iran

    A. Hedayati, S. Hemmaty & A. Amirsadeghi

  2. Department of Horticulture, Faculty of Agriculture, Urmia University, Urmia, Iran

    A. Hedayati & E. Nourozi

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  1. A. Hedayati
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  2. S. Hemmaty
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Correspondence to S. Hemmaty.

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Hedayati, A., Hemmaty, S., Nourozi, E. et al. Effect of Yeast Extract on h6h Gene Expression and Tropane Alkaloids Production in Atropa belladonna L. Hairy Roots. Russ J Plant Physiol 68, 102–109 (2021). https://doi.org/10.1134/S1021443721010088

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  • DOI: https://doi.org/10.1134/S1021443721010088

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