Abstract
Rosa rugosa Thunb. is a popular ornamental and medicinal plant native to eastern Asia. In this study, a successful bioreactor culture system was established for the production of secondary metabolites of rugosa roses. We tested different concentrations and combinations of plant hormones in growth media for maximum shoot proliferation and production of bioactive compounds, different bioreactor systems for maximum biomass production and production of bioactive compounds, and different ratios of nitrogen sources for maximum shoot growth and accumulation of bioactive compounds. For multiple shoot proliferation, Murashige and Skoog (MS) medium was used, supplemented with different concentrations and combinations of plant hormones: 6-benzylaminopurine (BA; 0-13.2 µM), thidiazuron (TDZ; 0-13.5 µM), and indole butyric acid (IBA) at 2.5 µM, used alone or in a combination of IBA with BA or TDZ. Rapid micropropagation of multiple shoots of rugosa roses was successfully achieved using shoot tips explanted in semisolid MS medium supplemented with 4.4 µM BA. The average number of shoots grown was 15.6 per explant and the maximum shoot length was 2.7 cm at 8 weeks of culture. To investigate the effect of nitrogen sources on shoot growth and bioactive compound accumulation, shoots were treated with different ratios of nitrogen sources (NH4 +:NO3 -) for 1 week after 7 weeks of shoot culture. Next, to scale up biomass production for the generation of useful phytochemicals, multiple-shoot cultures were developed in large-scale bioreactors. Four bioreactor systems were used: continuous immersion bioreactor (CIB), continuous immersion bioreactor with net (CIB-N), temporary immersion bioreactor (TIB), and temporary immersion bioreactor with net (TIB-N). Solid and liquid media were used as controls. Of the different bioreactor types, the CIB system produced the highest biomass, followed by the TIB system. Multiple shoots grown in the CIB system resulted in the accumulation of 39.21 mg·g-1 dry weight (DW) of total phenolics and 13.28 mg·g-1 DW of total flavonoids. The productivity of total phenolics and flavonoids was highest in the shoots harvested from the CIB system. The results of this study suggest that multiple shoots of rugosa roses can be used in commercial-scale bioreactors to produce useful bioactive compounds for the pharmaceutical and cosmetic industries.
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Literature Cited
Amid, A., N.N. Johan, P. Jamal, and W.N.W.M. Zain. 2013. Observation of antioxidant activity of leaves, callus and suspension culture of Justicia gendarusa. Afr. J. Biotechnol. 10;18653–18656.
Al Khateeb, W., E. Hussein, L. Qouta, M. Alu’datt, B. Al-Shara, and A. Abu-Zaiton. 2012. In vitro propagation and characterization of phenolic content along with antioxidant and antimicrobial activities of Cichorium pumilum Jacq. Plant Cell Tissue Organ Cult. 110;103–110.
Alvard, D., F. Cote, and C. Teisson. 1993. Comparison of methods of liquid medium culture for banana micropropagation. Effects of temporary immersion of explants. Plant Cell Tissue Organ Cult. 32;55–60.
Ara, K.A., M.M. Hossain, M.A. Quasim, M. Ali, and J.U. Ahmed. 1997. Micropropagation of rose (Rosa sp. cv). Plant Tissue Cult. 7;135–42.
Behrend, J. and R.I. Mateles. 1975. Nitrogen metabolism in plant cell suspension cultures I. Effect of amino acids on growth. Plant physiol. 56;584–589.
Carelli, B.P. and S. Echeverrigaray. 2002. An improved system for the in vitro propagation of rose cultivars. Sci. Hortic. 92;69–74.
Ciesla, L., J. Kryszen, A. Stochmal, W. Oleszeck, and M. Waksmundzka-Hajnos. 2012. Approach to develop a standardized TLC-DPPH test for assessing free radical scavenging properties of selected phenolic compounds. J. Pharma. Biomed. Anal. 70;126–135.
Czyzowska, A., E. Klewicka, E. Pogorzelski, and A. Nowak. 2015. Polyphenols, vitamin C and antioxidant activity in wines from Rosa canina L. and Rosa rugosa Thunb. J. Food Comp. Anal. 39;62–68.
Dai J. and R.J. Mumper. 2010. Plant phenolics: Extraction, analysis and their antioxidant and anticancer properties. Molecules 15; 7313–7352.
Dandin, V.S. and H.N. Murthy. 2012. Enhanced in vitro multiplication of Nothapodytes nimmoniana Graham using semisolid and liquid cultures and estimation of camptothecin in the regenerated plants. Acta Physiol. Plant 34;1381–1386.
Diwan, R., A. Shinde, and N. Malpathak. 2012. Phytochemical composition and antioxidant potential of Ruta graveolens L. in vitro culture lines. J. Bot. 2012:685427.
Ducos, J.P., B. Terrier, B. Coutois, and V. Petiard. 2008. Improvement of plastic based disposable bioreactors for plant science needs. Phytochem. Rev. 7;607–613.
Folin, O. and V. Ciocalteu. 1927. On trysonic and tryptophane determination in proteins. J. Biol. Chem. 27;627–650.
Hahn, E.J. and K.Y. Paek. 2005. Multiplication of Chrysanthemum shoots in bioreactors as affected by culture method and inoculation density of single node stems. Plant Cell Tissue Organ Cult. 81; 301–306.
Hashidoko, Y. 1996. The phytochemistry of Rosa rugosa. Phytochemistry 43;535–549.
Ibrahim, M.H., H.Z.E. Jaafar, A. Rahmat, and Z.A. Rahman. 2012. Involvement of nitrogen on flavonoids, glutathione, anthocyanin, ascorbic acid and antioxidant activities of Malaysian medicinal plant Labisia pumila Blume (Kacip Fatimah). Int. J. Mol. Sci. 13;393–408.
Ivanova, M. and J. Van Staden. 2009. Nitrogen source, concentration, and NH +4 : NO -3 ratio influence shoot regeneration and hyperhydricity in tissue cultured Aloe polyphylla. Plant Cell Tissue Organ Cult. 99;167–174.
Joo, S.S., Y.B. Kim, and D.I. Lee. 2010. Antimicrobial and antioxidant properties of secondary metabolites from white rose flower. Plant Pathol. J. 26;57–62.
Jung, H., J. Nam, J. Choi, K. Lee, and H. Park. 2005. 19a-hydroxyursane-type triterpenoids: Antiinociceptive anti-inflammatory principles of the roots of Rosa rugosa. Biol. Pharm. Bull. 28; 101–104.
Kaul, K. and S.A. Hoffman. 1993. Ammonium ion inhibition of Pinus strobus L. callus growth. Plant Sci. 88;169–173.
Konstas, J. and S. Kintzios. 2003. Developing a scale-up system for the micropropagation of cucumber (Cucumis sativus L.): the effect of growth retardants, liquid culture and vessel size. Plant Cell Rep. 21;538–548.
Krishnan, E.N., C.G. Sudha, and S. Seeni. 1995. Rapid propagation through shoot tip culture of Trichopus zeylanicus Gaetrn. a rare ethno medicinal plant. Plant Cell Rep. 14;708–711.
Krussmann G. 1982. Roses. B.T. Batsford Ltd., London. p. 269–272.
Liangxiong, X., Z. Youwei, L. Gang, and P. Yonghong. 2005. The antioxidant activities and their relationship with the relative polyphenols and flavonols contents of several flowers extracts. Chin. Wild Plant Res. 24;51–54.
Mark, T.R. and S.E. Simpson. 1994. Factors affecting shoot development in apically dominant Acer cultivars in vitro. J. Hortic. Sci. 69;543–551.
Miliauskas, G., P.R. Venskutonis, and T.A. van Beek. 2004. Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chem. 85;231–237.
Mimura, T., C. Shindo, M. Kato, E. Yokota, K. Sakano, H. Ashihara, and T. Shimmen. 2000. Regulation of cytoplasmic pH under extreme acid conditions in suspension cultured cells of Catharanthus roseus: a possible role of inorganic phosphate. Plant Cell Physiol. 41;424–431.
Naik, P.M., S.H. Manohar, H.N. Murthy. 2011. Effects of macro elements and nitrogen source on biomass accumulation and bacoside A production from adventitious shoot cultures of Bacopa monnieri (L.). Acta Physiol. Plant. 33;1553–1557.
Ng, T.B., J.S. He, S.M. Niu, L. Zhao, Z.F. Pi, W. Shao, and F. Liu. 2004. A gallic acid derivative and polysaccharides with antioxidative activity from rose (Rosa rugosa) flowers. J. Pharm. Pharmacol. 56;537–545.
Niemenak, N., K. Saare-Surminski, C. Rohsius, D.O. Ndoumou, and R. Lieberei. 2008. Regeneration of somatic embryos in Theobroma cacao L. in temporary immersion bioreactor and analyses of free amino acids in different tissues. Plant Cell Rep. 27;667–676.
Olech, M., R. Nowak, R. Los, J. Rzymowska, A. Malm, and K. Chrusciel. 2012. Biological activity and composition of teas and tinctures prepared from Rosa rugosa Thunb. Cent. Eur. J. Biol. 7;172–182.
Paek, K.Y., D. Chakrabarty, and E.J. Hahn. 2005. Application of bioreactor systems for large scale production of horticultural and medicinal plants, p. 95–116. In: Hvoslef-Eide, A.K., and W. Preil. (eds.). Liquid Culture System for in vitro Plant Propagation. Springer.
Park, S.Y., W.Y. Lee, J.K. Ahn, Y.J. Kwon, and H.C. Park. 2004. High frequency bioreactor culture system for mass proliferation and bulblet formation of Allium victorialis var platyphyllum Makino. Korean J. Plant Biotechnol. 31;127–132.
Park, S.Y. and K.Y. Paek. 2014. Bioreactor culture of shoots and somatic embryos of medicinal plants for production of bioactive compounds, p. 337–368. In: K.Y., Paek, H.N. Murthy, and J.J. Zhong (eds.). Production of biomass and bioactive compounds using bioreactor technology. Springer.
Park, J.A., B.J. Park, A.H. Kim, S.Y. Park, and K.Y. Paek. 2015. Airlift bioreactor system and nitrogen sources for biomass and antioxidant compound production from in vitro culture of Vitis flexuosa plantlets. Hortic. Environ. Biotechnol. 56;358–365.
Park, J.C., S.C. Kim, M.R. Choi, S.H. Song, E.J. Yoo, S.H. Kim, H. Miyashro, and M. Hattori. 2005. Anti-HIV protease activity from rosa family plant extracts and rosamultin from Rosa rugosa. J. Med. Food 8;107–109.
Praveen, N., N.P.M Naik, A. Manohar, S.H. Nayeem, and H.N. Murthy. 2009. In vitro regeneration of brahmi shoots using semisolid and liquid cultures and quantitative analysis of bacoside A. Acta. Physiol. Plant. 31;723–728.
Piatczak, E., I. Grzegorczyk-Karolak, and H. Wysokinska. 2014. Micropropagation of Rehmannia glutinosa Libosch: production of phenolics and flavonoids and evaluation of antioxidant activity. Acta Physiol. Plant. 36;1693–1702.
Sajc, L., D. Grubisic, and G. Vunjac-Novakovic. 2000. Bioreactors for plant engineering: an outlook for further research. Biochem. Bioeng. J. 4;89–99.
Sakanaka, S., Y. Tachibana, and Y. Okada. 2005. Preparation and antioxidant properties of extracts of Japanese persimmon leaf tea (kakinoha-cha). Food Chem. 89;569–575.
Sathyanarayana, B.N. and J. Blake. 1994. The effect of nitrogen sources and initial pH of the media with or without buffer on in vitro rooting of jackfruit (Artocarpus heterophyllus Lam.), p. 77–82. In: P.J., Lumsden, J.R. Nicholas, and W.J. Davies. (eds.). Physiology, Growth and Development of Plants in Culture. Springer Netherlands.
Verpoorte, R., A. Contin, and J. Memelink. 2002. Biotechnology for the production of plant secondary metabolites. Phytochemistry 1;13–25.
Wu, C.H., Y.H. Dewir, E.J. Hahn, and K.Y. Paek. 2006. Optimization of culturing conditions for the production of biomass and phenolics from adventitious roots of Echinacea angustifolia. J. Plant Biol. 49;193–199.
Xing, W., M. Bao, H. Qin, and G. Ning. 2010. Micropropagation of Rosa rugosa through axillary shoot proliferation. Acta Biol. Cracov. Bot. 52;69–75.
Yin, S., Y. Liang, W. Gao, J. Wang, S. Jing, Y. Zhang, and H. Liu. 2013. Influence of medium salt strength and nitrogen source on biomass and metabolite accumulation in adventitious root cultures of Pseudostellaria heterophylla. Acta Physiol. Plant. 35;2623–2628.
Youwei, Z. and P. Yonghong. 2007. Changes in antioxidant activity in Rosa rugosa flowers at different stages of development. N. Z. J. Crop Hortic. Sci. 35;397–401.
Yoshizawa, Y., S. Kawaii, M. Urashima, T. Fukase, T. Sato, R. Tanaka, N. Murofushi, and H. Nishimura. 2000. Antiproliferative effects of small fruit juices on several cancer cell lines. Anticancer Res. 20;4285–4289.
Zhong, J.J. and S.J. Wang. 1998. Effects of nitrogen source on the production of ginseng saponin and polysaccharide by cell cultures of Panax quinquefolium. Process Biochem. 33;671–675.
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Jang, HR., Lee, HJ., Shohael, A.M. et al. Production of biomass and bioactive compounds from shoot cultures of Rosa rugosa using a bioreactor culture system. Hortic. Environ. Biotechnol. 57, 79–87 (2016). https://doi.org/10.1007/s13580-016-0111-z
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DOI: https://doi.org/10.1007/s13580-016-0111-z