Maria PETROVA, Ely ZAYOVA, İvayla DINCHEVA, İlian BADJAKOV, Mariana VLAHOVA

Influence of carbon sources on growth and GC-MS based metabolite profiling of Arnica montana L. hairy roots

Arnica montana L. (Asteraceae) is an economically important herb that contains numerous valuable biologically active compounds accumulated in various parts of the plant. The effects of carbon sources (sucrose, maltose, and glucose) at different concentrations (1%, 3%, 5%, 7%, and 9%) on growth were studied and GC-MS based metabolite profiling of A. montana hairy roots was conducted. The optimal growth and biomass accumulation of transformed roots were observed on an MS nutrient medium containing 3% or 5% sucrose. GC-MS analysis of hairy roots of A. montana showed the presence of 48 compounds in polar fractions and 22 compounds in apolar fractions belonging to different classes of metabolites: flavones, phenolic acids, organic acids, fatty acids, amino acids, sugars, sugar alcohols, hydrocarbons etc. Among the various metabolites identified, only the sugars and sugar alcohols were influenced by the concentration of the respective carbon sources in the nutrient medium.

Anahtar Kelimeler:

Arnica, transformed roots, carbon sources, GC-MS analysis

Influence of carbon sources on growth and GC-MS based metabolite profiling of Arnica montana L. hairy roots

Keywords:

Arnica, transformed roots, carbon sources, GC-MS analysis,

PDF

___

Arsenault PR, Vail DR, Wobbe KK, Weathers PJ (2010). Effect of sugars on artemisinin production in Artemisia annua L.: transcription and metabolite measurements. Molecules 15: 2302–2318.
Bakkali AT, Jaziri M, Foriers A, Vander Heyden Y, Vanhaelen M, Homès J (1997). Lawsone accumulation in normal and transformed cultures of henna, Lawsonia inermis. Plant Cell Tiss Org 51: 83–87.
Borkowska B, Szezebra J (1991). Influence of different carbon sources on invertase-activity and growth of sour cherry (Prunus cerasus L.) shoot cultures. J Exp Bot 42: 911–915.
Cui Xi-Hua, Chakrabarty D, Lee Eun-Jung, Paek Kee-Yoeup (2010). Production of adventitious roots and secondary metabolites by Hypericum perforatum L. in a bioreactor. Bioresource Technol 101: 4708–4716.
Danphitsanuparn P, Boonsnongcheep P, Boriboonkaset T, Chintapakorn Y, Prathanturarug S (2012). Effects of Agrobacterium rhizogenes strains and other parameters on production of isoflavonoids in hairy roots of Pueraria candollei Grah. ex Benth. var. candollei. Plant Cell Tiss Org 111: 315–322.
Dupraz JM, Christen P, Kapetanidis I (1994). Tropane alkaloids in transformed roots of Datura quercifolia. Planta Med 60: 158– 162.
He HJ, Liang P, Shi HP (2005). Effects of sucrose and light on the growth and production of secondary metabolites in Pueraria phaseoloides hairy roots. Sheng Wu Gong Cheng Xue Bao 21: 1003–1008.
John KMM, Joshi SD, Mandal AKA, Kumar SR, Kumar RR (2009). Agrobacterium rhizogenes mediated hairy root production in tea leaves (Camellia sinensis (L.) O. Kuntze). Indian J Biotechnol 8: 430–434.
Jung KH, Kwak SS, Kim SW, Lee H, Choi CY, Liu JR (1992). Improvement of the catharanthine productivity in hairy root cultures of Catharanthus roseus by using monosaccharides as a carbon source. Biotechnol Lett 14: 695–700.
Kochan E, Szymańska G, Szymczyk P (2014). Effect of sugar concentration on ginsenoside biosynthesis in hairy root cultures of Panax quinquefolium cultivated in shake flasks and nutrient sprinkle bioreactor. Acta Physiol Plant 36: 613–619.
Kopka J, Schauer N, Krueger S, Birkemeyer C, Usadel B, Bergmuller E, Dormann P, Weckwerth W, Gibon Y, Stitt M (2005). GMD@ CSB. DB: the Golm Metabolome Database. Bioinformatics 21: 1635–1638.
Kozai T (1991). Micropropagation under photoautotrophic conditions. In: Debergh PC, Zimmerman RH, editors. Micropropagation: Technology and Application. London, UK: Kluwer Academic Press, pp. 447–469.
Krapp A, Hofmann B, Schäfer C, Stitt M (1993). Regulation of the expression of rbcS and other photosynthetic genes by carbohydrates: a mechanism for the ‘sink regulation’ of photosynthesis. Plant J 3: 817–828.
Liang P, Shi HP, Qi Y (2004). Effect of sucrose concentration on the growth and production of secondary metabolites in Pueraria phaseoloides hairy roots. Shi Yan Sheng Wu Xue Bao 37: 384– 390.
Liu CZ, Wang YC, Zhao B, Guo C, Ouyang F, Ye HC, Li GF (1999). Development of a nutrient mist bioreactor for growth of hairy roots. In Vitro Cell Dev-Pl 35: 271–274.
Liu Q, Cui L, Guo Y, Ni X, Zhang Y, Kai G (2013). Optimization of nutritive factors in culture media for growth and tropane alkaloid production from Anisodus acutangulus hairy roots. JAPS 3: 001–004.
Merfort I, Wendisch D (1985). Flavonoids from Arnica montana and Arnica chamissonis. Planta Med 51: 136–138.
Merfort I, Wendisch D (1992). New flavonoid glycosides from Arnicae flos DAB 9. Planta Med 58: 355–357.
Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plantarum 15: 473–497.
Murthy HN, Dijkstra C, Paul A, White DA, Davey MR, Power JB, Hahn EJ, Paek KY (2008). Establishment of Withania somnifera hairy root cultures for the production of withanolide A. J Integr Plant Biol 50: 975–981.
Nagella P, Thiruvengadam M, Jung SJ, Murthy HN, Chung IM (2013). Establishment of Gymnema sylvestre hairy root cultures for the production of gymnemic acid. Acta Physiol Plant 35: 3067–3073.
Neto VBP, Otoni WC (2003). Carbon sources and their osmotic potential in plant tissue culture: does it matter? Sci Hortic 97: 193–202.
Nikiforova V, Kopka J, Tolstikov V, Fiehn O, Hopkins L, Hawkesford M. (2005). Systems rebalancing the metabolism in response to sulfur deprivation, as revealed by metabolome analysis of Arabidopsis plants. Plant Physiol 138: 304–318.
Pavlov A, Berkov S, Weber J, Bley TH (2009). Hyoscyamine biosynthesis in Datura stramonium hairy root in vitro systems with different ploidy levels. Appl Biochem Biotech 157: 210– 225.
Petrova M, Zayova E, Vlahova M (2013). Induction of hairy roots in Arnica montana L. by Agrobacterium rhizogenes. Cent Eur J Biol 8: 470–479.
Pljevljakušić D, Rančić D, Ristić M, Vujisić L, Radanović D, Dajić- Stevanović Z (2012). Rhizome and root yield of the cultivated Arnica montana L., chemical composition and histochemical localization of essential oil. Ind Crop Prod 39: 177–189.
Praveen N, Murthy HN (2012). Synthesis of withanolide A depends on carbon source and medium pH in hairy root cultures of Withania somnifera. Ind Crop Prod 35: 241–243.
Putalun W, Pimmeuangkao S, De-Eknamkul W, Tanaka H, Shoyama Y (2006). Sennosides A and B production by hairy roots of Senna alata (L.) Roxb. Z Naturforsch C. 61: 367–371.
Romero FR, Delate K, Kraus GA, Solco AK, Murphy PA, Hannapel DJ (2009). Alkamide production from hairy root cultures of Echinacea. In Vitro Cell Dev-Pl 45: 599–609.
Rossetti V, Lombard A, Sancin P, Buffa M, Di Stefano R (1984). Characterization of Arnica montana dried roots. Int J Crude Drug Res 22: 53–60.
Schmidt TJ, Stausberg S, von Raison J, Berner M, Willuhn G (2006). Lignans from Arnica species. Nat Prod Res 20: 443–453.
Shakeran Z, Keyhanfar M, Asghari G, Ghanadian M (2015). Improvement of atropine production by different biotic and abiotic elicitors in hairy root cultures of Datura metel. Turk J Biol 39: 111–118.
Sivakumar G, Yu KW, Hahn EJ, Paek KY (2005). Optimization of organic nutrients for ginseng hairy roots production in large- scale bioreactors. Curr Sci India 89: 641–649.
Sivanesan I, Jeong BR (2009). Induction and establishment of adventitious and hairy root cultures of Plumbago zeylanica L. Afr J Biotechnol 8: 5294–5300.
Slesak H, Skoczowski A, Przywara L (2004). Exogenous carbohydrate utilisation by explants of Brassica napus cultured in vitro. Plant Cell Tiss Org 79: 45–51.
Srinivasan V, Pestchanker L, Moser S, Hirasuna TJ, Taticek RA, Shuler ML (1995). Taxol production in bioreactors: kinetics of biomass accumulation, nutrient uptake, and taxol production by cell suspension of Taxus baccata. Biotechnol Bioeng 47: 666–676.
Thiruvengadam M, Praveen N, John KM, Yang YS, Kim SH, Chung IM (2014). Establishment of Momordica charantia hairy root cultures for the production of phenolic compounds and determination of their biological activities. Plant Cell Tiss Org 118: 545–557.
Thompson M, Thorpe T (1987). Metabolic and non-metabolic roles of carbohydrates. In: Bonga JM, Durzan DJ, editors. Cell and Tissue Culture in Forestry. Dordrecht, the Netherlands: Martinus Nijhoff Publishers, pp. 89–112.
Udomsuk L, Jarukamjorn K, Tanaka H, Putalun W (2009). Isoflavonoid production in a hairy roots culture of Pueraria candollei. Z Naturforsch C 64: 687–691.
Weremczuk-Jezyna I, Kalemba D, Wysokinska H (2011). Constituents of the essential oil from hairy roots and plant roots of Arnica montana. J Essent Oil Res 23: 91–97.
Weremczuk-Jeżyna I, Kisiel W, Wysokińska H (2006). Thymol derivatives from hairy roots of Arnica montana. Plant Cell Rep 25: 993–996.
Willuhn G (1972a). Studies on the contents of Arnica species. V. Content and variations in content of volatile oils in the various organs of Arnica species. Planta Med 21: 221–245.
Willuhn G (1972b). Studies on the components of Arnica species. VI. Characterization and preparative separation of volatile oils from roots, rhizoma, leaves, and flower heads of various Arnica species. Planta Med 21: 329–342.
Willuhn G (1998). Arnica flowers: pharmacology, toxicology, and analysis of the sesquiterpene lactones-their main active substances. In: Lawson LD, Bauer R, editors. Phytomedicines of Europe – chemistry and biological activity. Washington DC: Am Chem. Soc 691, pp. 118–132.
Xu H, Park JH, Kim YK, Park NI, Lee SY, Park SU (2009). Optimization of growth and pyranocoumarins production in hairy root culture of Angelica gigas Nakai. J Med Plants Res 3: 978–981.