Abstract
The effect of 6 years of cultivation and use of table-sugar (TS) on the biomass/terpene alkaloid productivities and rol gene expression were studied in a hairy root (HR) clone of Rauvolfia serpentina. The media cost could be reduced >94 % by replacing sucrose (SUC) with TS—an unexplored avenue for HR cultivation. The overall productivities increased over long-term cultivation with sugar proving superior to SUC for biomass (24.4 ± 2.11 g/l DW after 40 days to 17.31 % higher) and reserpine (0.094 ± 0.008 % DW after 60 days to 193.8 % more) production. The latter however revealed comparatively better yields concerning ajmaline (0.507 ± 0.048 % DW after 60 days to 61.98 % higher) and yohimbine (0.628 ± 0.062 % DW after 60 days to 38.32 % higher), respectively. PCR amplification of rol genes confirmed long-term expression stability.
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Alpizar E, Dechamp E, Lapeyre-Montes F, Guilhaumon C, Bertrand B, Jourdan C, Lashermes P, Etienne H (2008) Agrobacterium rhizogenes-transformed roots of coffee (Coffea arabica): conditions for long-term proliferation and morphological and molecular characterization. Ann Bot 101:929–940
Ayadi R, Tremouillaux-Guiller J (2003) Root formation from transgenic calli of Ginkgo biloba. Tree Physiol 23:713–718
Baiza AM, Quiroz-Moreno A, Ruiz JA, Loyala-vargas VM (1999) Genetic stability of hairy root cultures of Datura stramonium. Plant Cell Tissue Organ Cult 59:9–17
Bulgakov VP (2008) Functions of rol genes in plant secondary metabolism. Biotechnol Adv 26:318–324
Chandra S (2012) Natural plant genetic engineer Agrobacterium rhizogenes: role of T-DNA in plant secondary metabolism. Biotechnol Lett 34:407–415
Dey A, De JN (2011) Ethnobotanical aspects of Rauvolfia serpentina L. Benth ex Kurz. in India, Nepal and Bangladesh. J Med Plant Res 5:144–150
Geerlings A, Hallard D, Martinez CA, Lopes CI, Heijden RV, Verpoorte R (1999) Alkaloid production by a Cinchona officinalis ‘Ledgeriana’ hairy root culture containing constitutive expression constructs of tryptophan decarboxylase and strictosidine synthase cDNAs from Catharanthus roseus. Plant Cell Rep 18:191–196
Guivarc’h A, Boccara M, Prouteau M, Chriqui D (1999) Instability of phenotype and gene expression in long-term culture of carrot hairy root clones. Plant Cell Rep 19:43–50
Hanisch ten cate CH, Loonen AEHM, Ottaviani MP, Ennik L, Eldik GV, Stiekema WJ (1990) Frequent spontaneous deletions of Ri T-DNA in Agrobacterium rhizogenes transformed potato roots and regenerated plants. Plant Mol Biol 14:735–741
Lipp-Joao KH, Brown TA (1994) Long-term stability of root cultures of tomato transformed with Agrobacterium rhizogenes R1601. J Exp Bot 45:641–647
Madhusudanan KP, Banerjee S, Khanuja SPS, Chattopadhyay SK (2008) Analysis of hairy root culture of Rauvolfia serpentina using direct analysis in real time mass spectrometric technique. Biomed Chromatogr 22:596–600
Maldonado-Mendoza IE, Ayora-Talavera T, Loyola-Vargas VM (1993) Establishment of hairy root cultures of Datura stramonium. Characterization and stability of tropane alkaloid production during long periods of sub-culturing. Plant Cell Tissue Organ Cult 33:321–329
Marconi PL, Setten LM, Calcena EN, Alvarez MA, Pitta-Alvarez SI (2008) Changes in growth and tropane alkaloid production in long-term culture of hairy roots of Brugmansia candida. Electron J Integr Biosci 3:38–44
Matsuura HN, Rau MR, Fett-Neto AG (2013) Oxidative stress and production of bioactive monoterpene indole alkaloids: biotechnological implications. Biotechnol Lett 36(2):191–200. doi:10.1007/s10529-013-1348-6
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Peebles CAM, Sander GW, Li M, Shanks JV, San KY (2009) Five year maintenance of the inducible expression of anthranilate synthase in Catharanthus roseus hairy roots. Biotechnol Bioeng 102:1521–1525
Santos PM, Figueiredo AC, Oliveira MM, Barroso JG, Pedro LG, Deans SG, Younus AKM, Scheffer JJC (1999) Morphological stability of Pimpinella anisum hairy root cultures and time-course study of their essential oils. Biotechnol Lett 21:859–864
Sevon N, Hiltunen R, Oksman-Caldentey KM (1998) Somaclonal variation in transformed roots and protoplast-derived hairy root clones of Hyoscyamus muticus. Planta Med 64:37–41
Sudha CG, Obul Reddy B, Ravishankar GA, Seeni S (2003) Production of ajmalicine and ajmaline in hairy root cultures of Rauvolfia micrantha Hook f., a rare and endemic medicinal plant. Biotechnol Lett 25:631–636
Zdravkovic-Korac S, Calic D, Druart PH, Radojevic L (2003) The horse chestnut lines harboring the rol genes. Biol Plant 47:487–491
Acknowledgments
The authors wish to express their sincere thanks to the Director, CSIR-CIMAP, for providing the facilities to carry out this research. Thanks are also due to Council of Scientific & Industrial Research (CSIR), New Delhi, India, for financial support to PP, RK and SS in the form of fellowships.
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Pandey, P., Kaur, R., Singh, S. et al. Long-term stability in biomass and production of terpene indole alkaloids by hairy root culture of Rauvolfia serpentina and cost approximation to endorse commercial realism. Biotechnol Lett 36, 1523–1528 (2014). https://doi.org/10.1007/s10529-014-1495-4
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DOI: https://doi.org/10.1007/s10529-014-1495-4