Abstract
These studies report the development of an efficient technique for large-scale cultivation of fast-growing hairy root culture systems for production of bioactive isoflavones. Trifolium pratense L. is an important source of pharmaceutically important isoflavones with immense health care applications. Trifolium pratense was transformed using different strains of Agrobacterium rhizogenes for hairy root induction and establishment of hairy root rhizoclones. Selected fast-growing rhizoclones of T. pratense were evaluated for their growth and isoflavone production. This study is the first report of stable production of isoflavones through successive culture passages from transformed hairy-root rhizoclones of T. pratense. One of the fast-growing hairy-root rhizoclones 2364A displayed significantly higher accumulation of all four pharmaceutically important isoflavones, 8.56 mg (gdw)−1 of daidzein, 2.45 mg (gdw)−1 of genistein, 15.23 mg (gdw)−1 of formononetin, and 1.10 mg (gdw)−1 of biochanin A, compared to other rhizoclones.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abdullah BM, Sharma P, Bernards MA, Guiller JT (2013) Phytochemicals, plant growth, and the environment. In: Gang DR (ed) Recent advances in Phytochemistry 42. Springer Science Business Media, New York, pp 95–142
Ali SS, Kasoju N, Luthra AL, Singh A, Sharanabasava H, Sahu H (2008) Indian medicinal herbs as sources of antioxidants. Food Res Int 41(1):1–15. https://doi.org/10.1016/j.foodres.2007.10.001
Anil Kumar M, Sravanthi Pammi SS, Giri A (2011) Establishment of suspension culture and production of isoflavones from red clover (Trifolium pratense L.) Int J App Biotechnol Biochem 1:35–42
Anil Kumar M, Vishnu Prasanth V, Giri A (2016) A validated RP-HPLC method for simultaneous determination of daidzein, genistein, formononetin and biochanin A from In Vitro cultured cells of Trifolium pratense L (Fabaceae). Der Pharma Chemica 8:16–21
Bertani G (1951) Studies on lysogenesis: the mode of phage liberation by lysogenic Escherichia coli. J Bacteriol 62(3):293–300
Bourgaud F, Gravot A, Milesi S, Gontier E (2001) Production of plant secondary metabolites: a historical perspective. Plant Sci 161(5):839–851. https://doi.org/10.1016/S0168-9452(01)00490-3
Dudareva NN, Klempien A, Muhlemann JK, Kaplan I (2013) Biosynthesis, function and metabolic engineering of plant volatile organic compounds. New Phytol 198(1):16–32. https://doi.org/10.1111/nph.12145
Estrada KR, Heriberto VL, Diego H, Elisabeth M, Rosa MC, Javier P (2016) Elicitation, an effective strategy for the biotechnological production of bioactive high-added value compounds in plant cell factories. Molecules 21(2):182. https://doi.org/10.3390/molecules21020182
Fahy PC, Hayward AC (1983) Media and methods. In: Fahy PC, Persley GJ GJ (eds) Plant Bacterial Diseases. Academic Press, Sydney, pp 337–378
Fu CX, Zhao DX, Xue XF, Jin ZP, Ma FS (2005) Transformation of Saussurea involucrate by Agrobacterium rhizogenes: hairy root induction and syringing production. Process Biochem 40:3789–3794
Georgiev MI, Agostini J, Müller JL, Xu J (2012) Genetically transformed roots: from plant disease to biotechnology. Trends Biotechnol 30(10):528–537. https://doi.org/10.1016/j.tibtech.2012.07.001
Giri A, Narasu ML (2000) Transgenic hairy roots: recent trends and applications. Biotechnol Adv 18:1–22
Giri CC, Zaheer M (2016) Chemical elicitors versus secondary metabolite production in vitro using plant cell, tissue and organ cultures: recent trends and a sky eye view appraisal. Plant Cell Tissue Organ Cult 126(1):1–18. https://doi.org/10.1007/s11240-016-0985-6
Giri A, Ravindra ST, Dingra V, Narasu ML (2001) Influence of different strains of Agrobacterium rhizogenes on induction of hairy roots and artimisinin production in Artemisia annua. Curr Sci 81:378–382
Guillon S, Trémouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006a) Harnessing the potential of hairy roots: dawn of a new era. Trends Biotechnol 24(9):403–409. https://doi.org/10.1016/j.tibtech.2006.07.002
Guillon S, Trémouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006b) Hairy root research: recent scenario and exciting prospects. Curr Opin Plant Biol 9(3):341–346. https://doi.org/10.1016/j.pbi.2006.03.008
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(6):1003–1008
Kaurinovic B, Popovic M, Vlaisavljevic SH, Schwartsova M, Vojinovic M (2012) Antioxidant profile of Trifolium pratense L. Molecules 17(12):11156–11172. https://doi.org/10.3390/molecules170911156
Kim YB, Reed DW, Covello PS (2015) Production of triterpenoid sapogeninsin hairy root cultures of Silene vulgaris. Nat Prod Commun 10(11):1919–1922
Kiranmayee R, Bhuvaneswari C, Narasu ML, Giri A (2012) Agrobacterium-mediated transformation in Alpinia galanga (Linn.) wild. for enhanced acetoxychavicol acetate production. Appl Biochem Biotechnol 168:339–347
Klejdus B, Sterbova D, Stratil P, Kuban V (2003) Identification and characterization of isoflavones in plant material by HPLC-DAD-MS tandem. Chem List 97:530–539
Kokotkiewicz A, Luczkiewicz M, Kowalski W, Badura A, Piekus N, Bucinsk A (2013) Isoflavone production in Cyclopia subternata Vogel (honeybush) suspension cultures grown in shake flasks and stirred-tank bioreactor. Appl Microbiol Biotechnol 97(19):8467–8477. https://doi.org/10.1007/s00253-013-5099-z
Kolodziejczyk-Czepas J (2012) Trifolium species-derived substances and extracts—biological activity and prospects for medicinal applications. J Ethnopharmacol 143(1):14–23. https://doi.org/10.1016/j.jep.2012.06.048
Luczkiewicz M, Kokotkiewicz A (2012) Elicitation and permeabilization affect the accumulation and storage profile of phytoestrogens in high productive suspension cultures of Genista tinctoria. Acta Physiol Plant 4:1–16
Majumdar S, Garai S, Jha S (2012) Use of the cryptogein gene to stimulate the accumulation of bacopa saponins in transgenic Bacopa monnieri plants. Plant Cell Rep 31(10):1899–1909. https://doi.org/10.1007/s00299-012-1303-3
Marisol OV, Susan H, Sun MH, Jang MO, Jin YW, Eun-Kyong Lee E, Gary J, Loake GJ (2016) Plant cell culture strategies for the production of natural products. BMB Rep 49:149–158
Müller JL, Linda J, Annemarie L, Joachim P, Antje W (2014) Improvement of hairy root cultures and plants by changing biosynthetic pathways leading to pharmaceutical metabolites: strategies and applications. Biotechnol Adv 32(6):1168–1179. https://doi.org/10.1016/j.biotechadv.2014.03.007
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
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(10):3067–3073. https://doi.org/10.1007/s11738-013-1327-5
Ooi CT, Syahida A, Stanslas J, Maziah M (2013) Efficiency of different Agrobacterium rhizogenes strains on hairy roots induction in Solanum mammosum. World J Microbiol Biotechnol 29(3):421–430. https://doi.org/10.1007/s11274-012-1194-z
Patisaul HB, Jefferson W (2010) The pros and cons of phytoestrogens. Front Neuroendocrinol 31(4):400–419. https://doi.org/10.1016/j.yfrne.2010.03.003
Pollier J, Morreel K, Geelen D, Goossens AJ (2011) Metabolite profiling of triterpene saponins in Medicago truncatula hairy roots by liquid chromatography Fourier transform ion cyclotron resonance mass spectrometry. Nat Prod 74(6):1462–1476. https://doi.org/10.1021/np200218r
Prathunturarug S, Schaffner W, Berger BK, Pank F (2007) Variation in growth and diterpene lactones among field-cultivated Andrographis paniculata. J Nat Med 61(2):159–163. https://doi.org/10.1007/s11418-006-0115-6
Rassi CM, Lieberherr M, Chaumaz G, Pointillart A, Cournot GJ (2002) Downregulation of osteoclast differentiation by daidzein via caspase3. Bone Miner Res 17(4):630–638. https://doi.org/10.1359/jbmr.2002.17.4.630
Ravindranath MH, Muthugounder S, Presser N, Viswanathan S (2004) Anticancer therapeutic potential of soy isoflavone genistein. In: Cooper EL, Yamaguchi N (eds) Complementary and alternative approaches to biomedicine. Advances in Experimental Medicine and Biology, vol 546. Springer, Boston. https://doi.org/10.1007/978-1-4757-4820-8_11
Ricigliano V, Javed C, Jingjing T, Andrew A, Dianella GH (2015) Plant hairy root cultures as plasmodium modulators of the slime mold emergent computing substrate Physarum polycephalum. Front Microbiol 6:1–10
Saviranta NMM, Anttonen MJ, Von Wright A, Karjalainen RO (2008) Red clover (Trifolium pratense L.) isoflavones: determination of concentrations by plant stage, flower colour, plant partand cultivar. J Sci Food Agric 88(1):125–132. https://doi.org/10.1002/jsfa.3056
Saviranta NMM, Julkunen-Tiitto R, Oksanen O, Karjalainen RO (2010) Leaf phenolic compounds in red clover (Trifolium pratense L.) induced by exposure to moderately elevated ozone. Environ Pollut 158(2):440–446. https://doi.org/10.1016/j.envpol.2009.08.029
Sharma V, Goyal S, Ramawat KG (2009) Scale up production of isoflavonoids in cell suspension cultures of Pueraria tuberosa grown in shake flasks and bioreactor. Eng Life Sci 9(3):267–271. https://doi.org/10.1002/elsc.200800114
Sheki H, Nishizawa T, Tanaka N, Niwa Y, Yoshida S, Muranaka T (2005) Hairy root-activation tagging: a high-throughput system for activation tagging in transformed hairy roots. Plant Mol Biol 59(5):793–807. https://doi.org/10.1007/s11103-005-1008-x
Shinde AN, Malpathak N, Fulzele DP (2009) Studied enhancement strategies for phytoestrogens production in shake flasks by suspension culture of Psoralea corylifolia. Bioresour Technol 100(5):1833–1839. https://doi.org/10.1016/j.biortech.2008.09.028
Swain SS, Sahu L, Barik DP, Chand PK (2010) Agrobacterium x plant factors influencing transformation of Joseph coat (Amarnthus tricolour L.) Sci Hortic 125(3):461–468. https://doi.org/10.1016/j.scienta.2010.04.034
Terrier B, Courtois D, Hénault N, Cuvier A, Bastin M, Aknin A, Dubreuil J, Pétiard V (2007) Two new disposable bioreactors for plant cell culture: the wave and undertow bioreactor and the slug bubble bioreactor. Biotechnol Bioeng 96(5):914–923. https://doi.org/10.1002/bit.21187
Thilip C, Raju CS, Varutharaju K, Aslam A, Shajahan A (2015) Improved Agrobacterium rhizogenes-mediated hairy root culture system of Withania somnifera (L.) Dunal using sonication and heat treatment. 3 Biotech 5:949–956
Thwe A, Mariadhas VA, Xiaohua L, Chang HP, Sun JK, Naif AAD, Sang UP (2016) Effect of different Agrobacterium rhizogenes strains on hairy root induction and phenylpropanoid biosynthesis in tartary buckwheat (Fagopyrum tataricum Gaertn). Front Microbiol 7:318–328. https://doi.org/10.3389/fmicb.2016.00318
Tusevski O, Stanoeva JP, Stefova M, Simic SG (2013) Phenolic profile of dark grown & photoperiod exposed Hypericum perforatum L. hairy root cultures. Sci World J 2013:602752. https://doi.org/10.1155/2013
Verma PC, Singh H, Negi AS, Saxena G, Rahman LU, Banerjee S (2015) Yield enhancement strategies for the production of picroliv from hairy root culture of Picrorhiza kurroa Royle ex Benth. Plant Signal Behav 10:1–11
Wei HC, Cai QY, Rahn RO (1996) Inhibition of UV light- and fenton reaction-induced oxidative DNA damage by the soybean isoflavone genistein. Carcinogenesis 17(1):73–77. https://doi.org/10.1093/carcin/17.1.73
Wu QL, Wang M, Simon JE (2003) Determination of isoflavones in red clover and related species by high-performance liquid chromatography combined with ultraviolet and mass spectrometric detection. J Chromatogr 1016(2):195–209. https://doi.org/10.1016/j.chroma.2003.08.001
Wu QL, Yang YH, Simon J (2011) Chemical profiling and quantification of isoflavone phytoestrogens in kudzu using LC/UV/MSD. Am J Anal Chem 2:665–674
Author information
Authors and Affiliations
Corresponding author
Additional information
Editor: Ming Cheng
Rights and permissions
About this article
Cite this article
Anil Kumar, M., Sravanthi Pammi, S.S., Sukanya, M.S. et al. Enhanced production of pharmaceutically important isoflavones from hairy root rhizoclones of Trifolium pratense L.. In Vitro Cell.Dev.Biol.-Plant 54, 94–103 (2018). https://doi.org/10.1007/s11627-017-9873-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11627-017-9873-y