GC-MS PROFILE OF IN VIVO, IN VITRO AND FUNGAL ELICITED IN VITRO LEAVES OF HYBANTHUS ENNEASPERMUS (L.) F. MUELL
Keywords:
Hybanthus enneaspermus, Suspension culture, In vivo plants-plants growing in the field, In vitro plants-plants raised through tissue culture technique in the laboratory, Fungal elicitors, GC-MSAbstract
Objectives: Investigation of the bioactive compounds from the methanol leaf extracts of in vivo, in vitro and fungal elicited in vitro plants of Hybanthus enneaspermus through GC-MS analysis.
Methods: The leaf explants were cultured on MS (Murashige and Skoog) medium supplemented with different concentrations of NAA (1-naphthalene acetic acid) for callus induction. The calli was treated with four different fungal elicitors, namely, Mucor prayagensis, Trichoderma viride, Fusarium moniliformis and Aspergillus niger on suspension culture containing the same growth regulators for two weeks. Regeneration of shoots was achieved from the fungal treated and untreated calli on MS medium fortified with different concentrations of cytokinins. Rooting was achieved from the isolated shoots on half strength MS medium containing different concentrations of auxins. The phytochemical composition was analyzed from the methanol extracts of in vivo, in vitro and fungal treated leaves of in vitro plants using gas chromatography and mass spectroscopy (GC-MS).
Results: Of the different concentrations and combinations of NAA, well developed green compact reproducible calli were obtained on MS medium supplemented with 10 µmol NAA+4 µmol BAP (6-benzylaminopurine). Shoots were regenerated from the fungal treated and untreated calli on MS medium containing 10 µmol BAP+6 µmol KIN (kinetin-6-furfurylaminopurine). Rooting was achieved on half strength MS medium supplemented with 9 µmol NAA. The GC-MS analysis revealed that the leaves of in vivo and in vitro plants contained 16 different phytochemicals, whereas, the fungal treated in vitro plants showed more number of phytochemicals, i.e., 22 (Mucor prayagensis), 26 (Trichoderma viride), 19 (Fusarium moniliformis) and 21 (Aspergillus niger) compounds.
Â
Â
Downloads
References
Canter PH, Thomas H, Ernst E. Bringing medicinal plants into cultivation: opportunities and challenges for biotechnology. Trends Biotechnol 2005;23:180–5.
Vijaya SN, Udayasri PV, Aswani KY, Ravi BB, Phani KY, Vijay VM. Advancements in the production of secondary metabolites. J Nat Prod 2010;3:112–23.
Sajc LD, Grubisic G, Vunjak N. Bioreactors for plant engineering: an outlook for further research. Biochem Eng J 2000;4:89–99.
Ataei-Azimi A, Delnavaz Hashemloian B, Ebrahimzadeh H, Majd A. High in vitro production of ant-canceric indole alkaloids from periwinkle (Catharanthus roseus) tissue culture. Afr J Biotechnol 2008;7:2834-9.
Deepika A, Vidya P, Uma K. In vitro propagation and quercetin quantification in callus cultures of Rasna (Pluchea lanceolata Oliver & Hiern). Indian J Biotechnol 2008;7:383-7.
Kumar V, Chauhan RS, Sood H. In vitro production and efficient quantification of major phytopharmaceuticals in an endangered medicinal herb, Swertia chirata. Int J Biotechnol Bioeng Res 2013;4:495-506.
Chen, Chia-Chen, Hung-Chi Chang, Chao-Lin Kuo, Dinesh Chandra Agrawal, Chi-Rei Wu, et al. In vitro propagation and analysis of secondary metabolites in Glossogyne tenuifolia (Hsiang-Ju)-a medicinal plant native to Taiwan. Botanical Studies 2014;55:1-9.
Brodelius P, Funk C, Haner A, Villegas M. A procedure for the determination of optimal chitosan concentrations for elicitation of cultured plant cells. Phytochemistry 1989;28:2651-4.
Zhao J, Davis L, Verpoorte R. Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnol Adv 2005;23:283–333.
Namdeo AG. Plant cell elicitation for production of secondary metabolites: a review. Pharmacogn Rev 2007;1:69–79.
Broeckling CD, Huhman DV, Farag MA, Smith JT, May GD, Pedro M, et al. Metabolic profiling of Medicago truncatula cell cultures reveals the effects of biotic and abiotic elicitors on metabolism. J Exp Bot 2005;56:323-36.
Hakkim FL, Kalyani S, Essa M, Girija S, Song H. Production of rosmarinic in Ocimum sanctum cell cultures by the influence of sucrose, phenylalanine, yeast extract, and methyl jasmonate. Int J Biol Med Res 2011;2:1070-4.
Devi CS, Nandi I, Mohana VS, Sriramkalyan P. Enhanced production of gymnemic acid using HR bioelicitor extracted from Xanthomonas spp. Int Res J Pharm 2012;3:221-5.
Ananthi P, Ranjithakumari BD. Gas chromatography-mass spectrometry analysis of methanolic seed and root extracts of Rorippa indica L. Int J ChemTech Res 2013;5:299-306.
Mahalakshmi R, Eganathan P, Parida AK. Salicylic acid elicitation on production of secondary metabolite by cell cultures of Jatropha curcas L. Int J Pharm Pharm Sci 2013;5:655-9.
Malini S, Eganathan P. GC-MS analysis of chemical composition of in vivo plant, in vitro and elicited roots of Bacopa monnieri (L.) Pennell. Anal Chem Lett 2013;3:380-8.
Namdeo AG, Patil S, Fulzele DP. Influence of fungal elicitors on production of ajmalicine by cell cultures of Catharanthus roseus. Biotechnol Prog 2002;18:159-62.
Ajungla L, Patil PP, Barmukh RB, Nikam TD. Influence of biotic and abiotic elicitors on accumulation of hyoscyamine and scopolamine in root cultures of Datura metel L. Indian J Biotechnol 2009;8:317–22.
Wiktorowska E, Dlugosz M, Janiszowska W. Significant enhancement of oleanolic acid accumulation by biotic elicitors in cell suspension cultures of Calendula officinalis L. Enzyme Microb Technol 2010;46:14–20.
Eilert U, Constabel F. Elicitation of sanguinarine accumulation in Papaver somniferum cells by fungal homogenates-An induction process. J Plant Physiol 1986;125:167-72.
Gadzovska-Simic SO, Antevski TS, Atanasova-Pancevska N, Petreska J, Stefova M, Kungulovski D, et al. Secondary metabolite production in Hypericum perforatum L. Cell Suspension upon elicitation with fungal mycelia from Aspergillus flavus. Arch Biol Sci Belgrade 2012;64:113-21.
Srinivasan V, Ciddi V, Bringi V, Shuler ML. Metabolic inhibitors, elicitors and precursors as tools for probing yield limitation in taxane production by Taxus chinensis cell cultures. Biotechnol Prog 1996;12:457-65.
Dixon RA, Achnine L, Kota P, Liu CJ, Reddy MKS, Wang L. The phenylpropanoid pathway and plant defence-a genomics perspective. Mol Plant Pathol 2002;3:371-90.
The Wealth of India. A dictionary of Indian Raw Materials and Industrial Products. Raw Materials Series, Publications and Information Directorate, CSIR, New Delhi; 1959;5:139.
Yoganarasimhan SN. In: Medicinal Plants of India-Tamilnadu, Cyber Media, Bangalore; 2000;2:276.
Hemalatha S, Wahi AK, Singh PN, Chansouria JPN. Anticonvulsant and free radical scavenging activity of Hybanthus enneaspermus: A preliminary screening. Indian J Traditional Knowledge 2003;2:383-8.
Thenmozhi TC, Premalakshmi V. Antioxidant effect of hydroethanolic extract of hybanthus enneaspermus on paracetamol induced oxidative stress in albino rats. Int J Res Pharm Biomed Sci 2011;2:1285-7.
Patel DK, Kumar R, Prasad SK, Sairam K, Hemalatha S. Antidiabetic and in vitro antioxidant potential of Hybanthus enneaspermus (Linn) F. Muell in streptozotocin-induced diabetic rats. Asian Pac J Trop Biomed 2011;1:316–22.
Weniger B, Lagnika L, Vonthron-Sénécheau C, Adjobimey T, Gbenou J, Moudachirou M, et al. Evaluation of ethnobotanically selected Benin medicinal plants for their in vitro antiplasmodial activity. J Ethnopharmacol 2004;90:279-84.
Tripathy S, Sahoo SP, Pradhan D, Sahoo S, Satapathy DK. Evaluation of anti arthritic potential of Hybanthus enneaspermus. Afr J Pharm Pharacol 2009;3:611-4.
Anand T, Gokulakrishnan K. GC–MS analysis and anti-microbial activity of bioactive components of Hybanthus enneaspermus. Int J Pharm Pharm Sci 2012;4:646-50.
Nandy S, Chakraborty B, Chatterjee P. Evaluation of anti-inflammatory activity of Hybanthus Enneaspermus Muell. leaves. Pharm Rev; 2012. p. 98-112.
Nathiya S, Senthamilselvi R. Anti infertility effect of Hybanthus enneaspermus on endosulfan induced toxicity in male rats. Int J Med Biosci 2013;2:28-32.
Setty MM, Narayanaswamy VB, Sreenivasan KK, Shirwaikar A. Free radical scavenging and nephroprotective activity of Hybanthus Enneaspermus (L) F. Muell. Pharmacologyonline 2007;2:158-71.
Velayutham P, Karthi C, Nalini P, Jahirhussain G. In vitro regeneration and mass propagation of Hybanthus enneaspermus (L.) F. Muell. from the stem explants through callus culture. J Agric Technol 2012;8:1119-28.
Murashige T, Skoog F. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 1962;15:473–97.
Staniszewska I, Krolicka A, Malinski E, Jojkowska E, Szafranek JZ. Elicitation of secondary metabolites in in vitro cultures of Ammi majus L., Enzy. Microbial Technol 2003;33:565-8.
Kim HJ, Sung MK, Kim JS. Anti-inflammatory effects of glyceollins derived from soybean by elicitation with Aspergillus sojae. Inflamm Res 2011;60:909–17.
Dr. Duke's Phytochemical and Ethnobotanical Databases. Available from: URL: http://www.ars-grin.gov/duke. [Last accessed on 10 May 2015]
Collin HA. Secondary product formation in plant tissue cultures. Plant Growth Regul 2001;34:119–34.
Mulabagal V, Tsay HS. Plant cell cultures-An alternative and efficient source for the production of biologically important secondary metabolites. Int J Appl Sci Eng 2004;2:29–48.
Martin EK, Vishal G, Susan CR. Pharmaceutically active natural product synthesis via plant cell culture technology. Mol Pharm 2008;5:243–56.
Karuppusamy S. A review on trends in production of secondary metabolites from higher plants by in vitro tissue, organ and cell cultures. J Med Plant Res 2009;3:1222–39.
Shalaka DK, Sandhya P. Micropropagation and organogenesis in Adhatoda vasica for estimation of vasine. Pharmacogn Mag 2009;5:359–63.
Facchini PJ, DeLuca VD. Opium poppy and madagascar periwinkle: model non-model systems to investigate alkaloid biosynthesis in plants. Plant J 2008;54:763–84.
Verma P, Mathur AK, Srivastava A, Mathur A. Emerging trends in research on spatial and temporal organization of terpenoid indole alkaloid pathway in Catharanthus roseus. Protoplasma 2012;249:255–68.
Harman GE, Howell CR, Viterbo A, Chet I, Lorito M. Trichoderma species—opportunistic, avirulent plant symbionts. Nat Rev Microbiol 2004;2:43–56.
Cordo CA, Monaco CL, Segarra CI, Simon MR, Mansilla AY, Perello AE, et al. Trichoderma spp. as elicitors of wheat plant defence responses against Septoria tritici. Biocontrol Sci Technol 2007;17:687–98.
Vinale F, Sivasithamparam K, Ghisalberti EL, Marra R, Barbetti MJ, Li H, et al. A novel role for Trichoderma secondary metabolites in the interactions with plants. Physiol Mol Plant Pathol 2008;72:80–6.
Archana P, Archana M, Alok K, Gupta MM, Lal RK, Mathur AK. Fungal elicitor-mediated enhancement in growth and asiaticoside content of Centella asiatica L. shoot cultures. Plant Growth Regul 2013;69:265–73.
Navazio L, Baldan B, Moscatiello R, Zuppini A, Woo SL, Mariani P, et al. Calcium-mediated perception and defense responses activated in plant cells by metabolite mixtures secreted by the biocontrol fungus Trichoderma atroviride. BMC Plant Biol 2007;7:41.