A strategy for overaccumulation of scopolamine in Datura innoxia hairy root cultures

• Autorzy: Dechaux C , Boitel-Conti M.
• Języki publikacji: EN

Abstrakty

EN

Hairy root culture plays an important role in investigation of alkaloid production in culture in vitro. The complexity of scientific work and the production capacity limitations associated with whole plant systems and unorganized cells can be overcome using culture of hairy roots. This paper presents an example of a strategy to produce secondary metabolites from plants, in which we investigated the alkaloid pathway, modified gene expression, and cultivated and optimized hairy root growth in a bioreactor. Datura innoxia whole plant was transformed by Agrobacterium rhizogenes, and the obtained hairy roots were studied for their tropane alkaloid production. Optimization of medium led to an increase of scopolamine production in hairy root culture of Datura innoxia by increasing the produced biomass, but genetic engineering seems to be the best way to increase the accumulation of scopolamine. These root cultures were not able to stably overaccumulate scopolamine. The involvement of putative enzymatic regulation of the tropane alkaloid pathway is discussed.

Słowa kluczowe

EN

tropane alkaloid; overaccumulation; strategy; alkaloid production; root; bioreactor; Datura innoxia; scopolamine; h6h gene; root culture; genetic engineering; hairy root; plant

• Wydawca: -
• Czasopismo: Acta Biologica Cracoviensia. Series Botanica
• Rocznik: 2005
• Tom: 47
• Numer: 1
• Opis fizyczny: p.101-107,fig.,ref.

Twórcy

autor

Dechaux C

• Universite de Picardie Jules Verne, 33, rue Saint Leu, F-80039 Amiens Cedex 01, France

autor

Boitel-Conti M.

Bibliografia

• Boitel-Conti M, Gontier E, Laberche JC, Ducrocq C, and Sangwan-Norreel BS. 1995. Permeabilization of Datura innoxia hairy roots for release of stored tropane alkaloids. Planta Medica 61: 287-290.
• Boitel-Conti M, Gontier E, Laberche JC, Ducrocq C, and Sangwan-Norreel BS. 1996. Inducer effect of Tween 20 permeabilization treatment used for release of stored tropane alkaloids in Datura innoxia Mill. Hairy roots cultures. Plant Cell Reports 16: 241-244.
• Boitel-Conti M, Laberche JC, Lanoue A, Ducrocq C, and Sangwan-Norreel BS. 2000. Influence of feeding precursors on tropane alkaloid production during an abiotic stress in Datura innoxia transformed roots. Plant Cell Tissue and Organ Culture 60: 131-137.
• Doran PM. 1997. Hairy roots, Culture and applications. Harwood Academic Publishers, Amsterdam.
• Giri A, and Narasu ML. 2000. Transgenic hairy roots: recent trends and applications. Biotechnology Advances 18: 1-22.
• Gontier E, Sangwan-Norreel BS, and Barbotin J-N. 1994. Simultaneous determination of some tropane alkaloids by reverse phase HPLC for Datura innoxia Mill. plant material. Agro-food Industry Hi-Tech 3: 26-28.
• Flores HE, Vivanco JM, and Loyola-Vargas VM. 1999. "Radicle” biochemistry: the biology of root-specific metabolism. Trends in Plant Science 4: 220-226.
• Hamill JD, Parr AJ, Rhodes MJC, Robins RJ, and Walton NJ. 1987. New routes to plant secondary products. Bio-Technology 5: 800-804.
• Hashimoto T, Matsuda J, and Yamada Y. 1993. Two-step epoxidation of hyoscyamine to scopolamine is catalyzed by bifunctional hyoscyamine 6b-hydroxylase. FEBS Letters 329: 35-39.
• Jouhikainen K, Lindgren L, Jokelainen T, Hiltunen R, Teeri TH, and Oksman-Caldentey KM. 1999. Enhancement of scopolamine production in Hyoscyamus muticus L. hairy root culture by genetic engineering. Planta 208: 545-551.
• Kamada H, Okamura N, Satake M, Harada H, and Shimomura K. 1986. Alkaloid Production by Hairy Root Cultures in Atropa belladonna. Plant Cell Reports 5: 239-242.
• Lanoue A, Boitel-Conti M, Laberche JC, Portais JC, Barbotin JN, Christen P, and Sangwan-Norreel BS. 2002. 13C-NMR study of hyoscyamine biosynthesis in Datura innoxia transgenic root cultures. Journal of Natural Products 65: 1131-1135.
• Lanoue A, Boitel-Conti M, Dechaux C, Laberche JC, Christen P, and Sangwan-Norreel BS. 2004. Comparison of growth properties, alkaloid production and water uptake of two selected Datura hairy roots lines. Acta Biologica Cracoviensia Series Botanica 46: 185-192.
• Matsuda J, Okabe S, Hashimoto T, and Yamada Y. 1991. Molecular cloning of hyoscyamine 6b-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, from cultured roots of Hyoscyamus niger. Journal of Biological Chemistry 266: 9460-9464.
• Moyano E, Fornale S, Palazon J, Cusido R M, Bagni N, and Pinol MT. 2002. Alkaloid production in Duboisia hybrid hairy root cultures overexpressing the pmt gene. Phytochemistry 59: 697-702.
• Moyano E, Jouhikainen K, Tammela P, Palazon J, Cusido R M, Pinol MT, Teeri TH, and Oksman-Caldentey KM. 2003. Effect of pmt gene overexpression on tropane alkaloid production in transformed root cultures of Datura metel and Hyoscyamus muticus. Journal of Experimental Botany 54: 203-11.
• Palazon J, Moyano E, Cusido RM, Bonfill M, Oksman-Caldentey KM, and Pinol MT. 2003. Alkaloid production in Duboisia hybrid hairy roots and plant overexpressing the h6h gene. Plant Science 165: 1289-1295.
• Ramachandra RAO S, and Ravishankar GA. 2002. Plant cell cultures: Chemical factories of secondary metabolites. Biotechnology Advances 20: 101-153.
• Rothe G, Hachiya A, Yamada Y, Hashimoto T, and Drager B. 2003. Alkaloids in plants and root cultures of Atropa belladonna overexpressing putrescine N-methyltransferase. Journal of Experimental Botany 54: 2065-70.
• Sato F, Hashimoto T, Hachiya A, Tamura K, Choi KB, Morishige T, Fujimoto H, and Yamada Y. 2001. Metabolic engineering of plant alkaloid biosynthesis. Proceedings of the National Academy of Sciences U.S.A. 98: 367-72.
• Shanks JV, and Morgan J. 1999. Plant "hairy root" culture. Current Opinion in Biotechnology 10: 151-155.
• Sporer F, Sauerwein M, and Wink M. 1993. Diurnal and developmental variation of alkaloid accumulation in Atropa belladonna. Acta Horticulturae 331: 381-386.
• Wilson PDG. 1997. The pilot-scale cultivation of transformed roots. In: Doran PM [ed.], Hairy roots, culture and applications, 179-190. Harwood Academic Publishers, Amsterdam.
• Yun DJ, Hashimoto T, and Yamada Y. 1992. Metabolic engineering of medicinal plants: transgenic Atropa belladonna with an improved alkaloid composition. Proceedings of the National Academy of Sciences U.S.A. 89: 11799-803.