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Influence of hairy root ecotypes on production of tropane alkaloids in Brugmansia candida

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Abstract

Hyoscyamine and scopolamine are tropane alkaloids widely applied in medicine. Differences in alkaloid production and growth kinetics have been observed in Argentinian and Colombian ecotypes of Brugmansia candida hairy roots. The aim of this work was to analyze the production of key intermediates in tropane alkaloid synthesis in both ecotypes to determine differences in the biosynthetic pathway. Additionally, rolC gene expression was analyzed to determine its correlation with hairy root growth. The results showed a higher accumulation of polyamines in Colombian hairy roots, suggesting that there may be a rate-limiting enzyme in the last steps of hyoscyamine biosynthesis. Additionally, rolC gene expression was correlated with an improvement in hairy root growth, which supports the function of rol genes as growth modulators and suggests that metabolic engineering approaches involving rolC manipulation may be useful for the development of more efficient B. candida hairy root cultures for biotechnological applications.

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References

  1. Altamura MM (2004) Agrobacterium rhizogenes rolB and rolD genes: regulation and involvement in plant development. Plant Cell Tissue Organ Cult 77:89–101

  2. Bulgakov VP (2008) Functions of rol genes in plant secondary metabolism. Biotechnol Adv 26:318–324. doi:10.1016/j.biotechadv.2008.03.001

  3. Cardillo AB, Rodriguez Talou J, Giulietti AM (2008) Expression of Brugmansia candida hyoscyamine 6beta-hydroxylase gene in Saccharomyces cerevisiae and its potential use as biocatalyst. Microb Cell Fact 7:17–23. doi:10.1186/1475-2859-7-17

  4. Cardillo AB, Otalvaro Alvarez AM, Calabro Lopez A, Velasquez Lozano ME, Rodriguez Talou J, Giulietti AM (2010a) Anisodamine production from natural sources: seedlings and hairy root cultures of argentinean and colombian Brugmansia candida plants. Planta Med 76:402–405. doi:10.1055/s-0029-1186164

  5. Cardillo AB, Otálvaro AM, Busto VD, Talou JR, Velásquez ME, Giulietti AM (2010b) Scopolamine, anisodamine and hyoscyamine production by Brugmansia candida hairy root cultures in bioreactors. Process Biochem 45:1577–1581

  6. Dehghan E, Hakkinen ST, Oksman-Caldentey KM, Ahmadi FS (2012) Production of tropane alkaloids in diploid and tetraploid plants and in vitro hairy root cultures of Egyptian henbane (Hyoscyamus muticus L.). Plant Cell Tissue Organ Cult 110:35–44. doi:10.1007/s11240-012-0127-8

  7. Di Rienzo JA, Casanoves F, Balzarini MG, Gonzalez L, Tablada M, Robledo CW (2010) InfoStat. Universidad Nacional de Córdoba, Córdoba

  8. Diwan R, Malpathak N (2008) Novel technique for scaling up of micropropagated Ruta graveolens shoots using liquid culture systems: a step towards commercialization. N Biotechnol 25:85–91. doi:10.1016/j.nbt.2008.02.002

  9. Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158. doi:10.1016/0014-4827(68)90403-5

  10. Giulietti AM, Parr AJ, Rhodes MJ (1993) Tropane alkaloid production in transformed root cultures of Brugmansia candida. Planta Med 59:428–431. doi:10.1055/s-2006-959725

  11. Grishchenko OV, Kiselev KV, Tchernoded GK, Fedoreyev SA, Veselova MV, Bulgakov VP, Zhuravlev YN (2013) The influence of the rolC gene on isoflavonoid production in callus cultures of Maackia amurensis. Plant Cell Tissue Organ Cult. doi:10.1007/s11240-012-0283-x

  12. Guillon S, Tremouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006) Hairy root research: recent scenario and exciting prospects. Curr Opin Plant Biol 9:341–346. doi:10.1016/j.pbi.2006.03.008

  13. Hakkinen ST, Moyano E, Cusido RM, Palazon J, Pinol MT, Oksman-Caldentey KM (2005) Enhanced secretion of tropane alkaloids in Nicotiana tabacum hairy roots expressing heterologous hyoscyamine-6beta-hydroxylase. J Exp Bot 56:2611–2618. doi:10.1093/jxb/eri253

  14. Hamill JD, Rounsley S, Spencer A, Todd G, Rhodes JC (1991) The use of the polymerase chain reaction in plant transformation studies. Plant Cell Rep 10:221–224

  15. Hashimoto T, Yamada Y (1986) Hyoscyamine 6β-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, in alkaloid-producing root cultures. Plant Physiol 81:619–625

  16. Hashimoto T, Yukimune Y, Yamada Y (1989) Putrescine and putrescine N-methyltransferase in the biosynthesis of tropane alkaloids in cultured roots of Hyocyamus albus I. Biochemical studies. Planta 178:123–130

  17. Jouhikainen K, Lindgren L, Jokelainen T, Hiltunen R, Teeri TH, Oksman-Caldentey KM (1999) Enhancement of scopolamine production in Hyoscyamus muticus L. hairy root cultures by genetic engineering. Planta 208:545–551

  18. Kursinszki L, Hank H, Laszlo I, Szoke E (2005) Simultaneous analysis of hyoscyamine, scopolamine, 6beta-hydroxyhyoscyamine and apoatropine in Solanaceous hairy roots by reversed-phase high-performance liquid chromatography. J Chromatogr A 1091:32–39

  19. Liu C (2008) Novel plant reactors for pharmaceutical production. In: World congress on in vitro biology, Arkansas State University. NewsRelease, University Communications Office. http://asunews.astate.edu/ABIRelease08.htm

  20. Liu T, Zhu P, Cheng KD, Meng C, Zhu HX (2005) Molecular cloning and expression of putrescine N-methyltransferase from the hairy roots of Anisodus tanguticus. Planta Med 71:987–989. doi:10.1055/s-2005-871260

  21. Moyano E, Fornale S, Palazon J, Cusido RM, Bagni N, Pinol MT (2002) Alkaloid production in Duboisia hybrid hairy root cultures overexpressing the pmt gene. Phytochemistry 59:697–702. doi:10.1016/S0031-9422(02)00044-4

  22. Onrubia M, Moyano E, Bonfill M, Palazon J, Goossens A, Cusido RM (2011) The relationship between TXS, DBAT, BAPT and DBTNBT gene expression and taxane production during the development of Taxus baccata plantlets. Plant Sci 181:282–287. doi:10.1016/j.plantsci.2011.06.006

  23. Palazon J, Cusido RM, Roig C, Piñol MT (1997) Effect of rol genes from Agrobacterium rhizogenes TL-DNA on nicotine production in tobacco root cultures. Plant Physiol Biochem 35:155–162

  24. Palazón J, Cusidó RM, Roig C, Piñol MT (1998) Expression of the rol C gene and nicotine production in transgenic roots and their regenerated plants. Plant Cell Rep 17:384–390

  25. Palazón J, Moyano E, Cusidó RM, Bonfill M, Oksman-Caldentey K, Piñol MT (2003) Alkaloid production in Duboisia hybrid hairy roots and plants overexpressing the h6h gene. Plant Sci 165:1289–1295

  26. Payne J, Hamill JD, Robins RJ, Rhodes MJ (1987) Production of hyoscyamine by ‘hairy root’ cultures of Datura stramonium. Planta Med 53:474–478. doi:10.1055/s-2006-962776

  27. Poupko JM, Baskin SI, Moore E (2006) The pharmacological properties of anisodamine. J Appl Toxicol 27:116–121. doi:10.1002/jat.1154

  28. Robins RJ, Bent EG, Rhodes MJC (1991) Studies on the biosynthesis of tropane alkaloids by Datura stramonium L. transformed root cultures. Planta 185:385–390. doi:10.1007/bf00201061

  29. Roses OE, Miño J, Villamil EC (1988) Acción farmacodinámica de las flores de Brugmansia candida. Fitoterapia 59:120–127

  30. Samuelsson G (ed) (1999) Drugs of natural origin, 4th edn. Gunnar Samuelsson and Apotekarsocieteten—Swedish Pharmaceutical Society, Swedish Pharmaceutical Press, Sweden

  31. Sato F, Hashimoto T, Hachiya A, Tamura K, Choi KB, Morishige T, Fujimoto H, Yamada Y (2001) Metabolic engineering of plant alkaloid biosynthesis. Proc Natl Acad Sci USA 98:367–372

  32. Sheng CY, Gao WY, Guo ZR, He LX (1997) Anisodamine restores bowel circulation in burn shock. Burns 23:142–146. doi:10.1016/S0305-4179(96)00086-1

  33. Smolka A, Li XY, Heikelt C, Welander M, Zhu LH (2010) Effects of transgenic rootstocks on growth and development of non-transgenic scion cultivars in apple. Transgenic Res 19:933–948. doi:10.1007/s11248-010-9370-0

  34. Spollansk TC, Pitta Alvarez SI, Giulietti AM (2000) Effect of jasmonic acid and aluminum on production of tropane alkaloids in hairy root cultures of Brugmansia candida. Electron J Biotechnol 3:72–75

  35. Taiz L, Zeiger E (2010) Plant physiology, 5th edn. Sinauer Associates, Inc., Sunderland

  36. Torrigiani P, Rabiti AL, Betti L, Marani F, Brizzi M, Bagni N, Canova A (1995) Improved method for polyamine determination in TMV, a rod-shaped virus. J Virol Methods 53:157–163. doi:10.1016/0166-0934(95)00008-I

  37. van der Salm TPM, van der Toorn CG, Bouwer R, Hanisch ten Cate CH, Dons HJM (1997) Production of ROL gene transformed plants of Rosa hybrida L. and characterization of their rooting ability. Mol Breed 3:39–47

  38. Wang H, Lu Y, Chen HZ (2003) Differentiating effects of anisodamine on cognitive amelioration and peripheral muscarinic side effects induced by pilocarpine in mice. Neurosci Lett 344:173–176. doi:10.1016/S0304-3940(03)00444-0

  39. Wu YF, Lü W, Lu Q, Zhang WS (2005) Asymmetric synthesis of anisodine. Chin Chem Lett 16:1287–1289

  40. Yamada Y, Yun DJ, Hashimoto T (1994) Genetic engineering of medicinal plants for tropane alkaloid production. In: Ryu DDY, Furusaki S (eds) Studies in plant sci. Advances in plant biotechnology, vol 4. Elsevier, Amsterdam, pp 83–93

  41. Zarate R, el Jaber-Vazdekis N, Medina B, Ravelo AG (2006) Tailoring tropane alkaloid accumulation in transgenic hairy roots of Atropa baetica by over-expressing the gene encoding hyoscyamine 6beta-hydroxylase. Biotechnol Lett 28:1271–1277. doi:10.1007/s10529-006-9085-8

  42. Zarate R, el Jaber-Vazdekis N, Cequier-Sanchez E, Gutierrez-Nicolas F, Ravelo AG (2008) Biotechnology for the production of plant natural products. Stud Nat Prod Chem 34:309–392

  43. Zhang L, Ding R, Chai Y, Bonfill M, Moyano E, Oksman-Caldentey KM, Xu T, Pi Y, Wang Z, Zhang H, Kai G, Liao Z, Sun X, Tang K (2004) Engineering tropane biosynthetic pathway in Hyoscyamus niger hairy root cultures. Proc Natl Acad Sci USA 101:6786–6791. doi:10.1073/pnas.0401391101

  44. Zhao B, Agblevor FA, Ritesh KC, Jelesko JG (2013) Enhanced production of the alkaloid nicotine in hairy root cultures of Nicotiana tabacum L. Plant Cell Tissue Organ Cult. doi:10.1007/s11240-012-0256-0

  45. Zia M, Mirza B, Malik SA, Chaudhary MF (2010) Expression of rol genes in transgenic soybean (Glycine max L.) leads to changes in plant phenotype, leaf morphology, and flowering time. Plant Cell Tissue Organ Cult 103:227–236. doi:10.1007/s11240-010-9771-z

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Acknowledgments

This work was supported by the program of short stays abroad for postdoctoral fellows from CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina), and the projects BIO2008-01210, BIO2011-29856-C02-01 (Ministerio de Ciencia e Innovación, España) and 2009SGR2013 (Generalitat de Catalunya). A.B.C. and A.M.G. are researchers from CONICET.

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Correspondence to Alejandra B. Cardillo.

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Cardillo, A.B., Giulietti, A.M., Palazón, J. et al. Influence of hairy root ecotypes on production of tropane alkaloids in Brugmansia candida . Plant Cell Tiss Organ Cult 114, 305–312 (2013). https://doi.org/10.1007/s11240-013-0326-y

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Keywords

  • Hyoscyamine
  • Scopolamine
  • 6β-Hydroxyhyoscyamine
  • Hyoscyamine-6β-hydroxylase
  • Polyamines
  • rolC