PLANT REGENERATION THROUGH IN VITRO AXILLARY MULTIPLICATION IN SHOOT TIPS AND COMPARATIVE STUDY OF ANTIOXIDANT DEFENSE SYSTEM IN ARTEMISIA NILAGIRICA (CLARKE)PAMP.

  • A.K. BALIARSINGH1*, L.SAMANTA 2 AND S.S.MOHANTY 3 1.BIOCHEMISTRY AND MOLECULAR BIOLOGY LABORATORY, P.G.DEPARTMENT OF BOTANY, UTKAL UNIVERSITY, VANI VIHAR, BHUBANESWAR-751004, ODISHA, INDIA. 2. P.G.DEPARTMENT OF ZOOLOGY, REVENSHAW UNIVERSITY, CUTTACK-753003, ODISHA, INDIA. 3.DEPARTMENT OF BOTANY, B.J.B. COLLEGE, BHUBANESWAR-751014, ODISHA, INDIA. Corresponding author’s e-mail: ajitonline75@gmail.com
Keywords: Artemisia nilagirica, In vitro axillary shoot proliferation, Antioxidants, Growth regulators.

Abstract

This paper describes a suitable protocol for in vitro axillary shoot proliferation and plant regeneration using shoot tip explants of an important medicinal and aromatic herb Artemisia nilagirica (Clarke)Pamp. Shoot apices were taken for axillary multiplication in MS medium supplemented with various growth regulators. Combinations of IAA (0.5 mg/L) with BAP (1.0 mg/L) in MS medium exhibited maximum organogenesis (shooting and rooting) after 10 weeks of culture. Plantlets emerged from culture medium were acclimatized and the survival rate of ex vitro plants after soil transplantation was 80-83% with no apparent phenotypic variations. A study was undertaken to compare levels of enzymatic and non-enzymatic antioxidants in natural in vivo plants and in vitro-regenerated plants before and after acclimatization (ex vitro). The total protein, ascorbic acid and phenolics contents as well as activities of antioxidative enzymes such as superoxide dismutase (SOD), catalase (CAT), peroxidise (POX), were found to be   maximum in ex vitro acclimatized plants than in vitro or in vivo plants respectively. Interestingly,  lipid peroxidation (LPx) and peroxide content were recorded at a  lower value in ex vitro acclimatized plants compared to in vitro grown and naturally occurring plants of A. nilagirica.

References

Aebi, H. (1984) Catalase. In: Methods in enzymology, L. Packer (Eds.), Academic Press, Orlando. 105: 121-126.
Ahameethunisa, A.R. and Hopper,W.(2010) Antibacterial activity of Artemisia nilagirica leaf extracts against clinical and phytopathogenic bacteria. BMC Complement Altern Med.106:10-6.
Aileni, M. Kota, S.R. Kokkirala, V.R. Umate, P. and Abbagani, S. (2009). Efficient In vitro regeneration and micropropation of medicinal plant Momordica tuberose Roxb. J. of Herbs, Spices & Med. plants, 15: 241-247.
Bowler, C. Van Montagu, M. and Inze, D. (1992) Superoxide dismutase and stress tolerance. Annu. Rev. Plant Physiol. Plant Mol. Biol. 43:83-116.
Das, K. L. Samanta, L. Chainy, G.B.N. A modified spectrophotometric assay of superoxide dismutase using nitrite formation by superoxide radicals. Ind. J. Biochem. Biophys. 37 (2000) 201–204.
Dev, S. (1997) Ethno therapeutics and modern drug development: The Potential of Ayurveda, Curr. Sci., 73(11): 909-928.
Elstner, E.F. (1982) Oxygen activation and oxygen toxicity. Annu. Rev. Plant Physiol. 33: 73-93.
Elstner, E.F. Wagner, G.A. and Schutz, W. (1988). Activated oxygen in green plants in relation to stress situations. Curr. Topics Plant Boichem. Physiol. 7: 159-187.
Ezenwaka, C. L. Mbagwee, F. N. and Unamba, C. I. N. (2009). Effect of combination of different Levels of Auxins and Cytokinines on in-vitro propagation of Dioscorea rotundata L.(White Yam), New York Science Journal, 2(5).
Govindaraj, S. (2008). Mass propagation and essential oil analysis of Artemisia vulgaris, J. of Biosc. and Bioengg. 105 (3): 176-183.
Halliwell, B. and Gutteridge, J.M.C. (1985) Free Radicals in Biology and Medicine. Clrendon Press, Oxford.
Heath, R. L. and Packer, L.(1968). Photooxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch. Biochem. Biophys.125:189-198.
Kar, M. and Feierabend, J. (1984) Metabolism of activated oxygen in detached wheat and rye leaves and its relevance to the initiation of senescence. Planta 160: 385-391.
Kishore, N. Dubey, N.K. and Chansouria, J.P.N. (2001). Antimycotic activity of the essentialoil of Artemisia nilagirica, Flavour and Fragrance Journal, 16 (1): 61-63.
Lowry,O.H. Rosebrough, N.J. Farr, A.L. and Randall, R.J.(1951) Protein measurement with Folin-phenol reagent. J.Biol.Chem.193:265-275.
Mitsui, A. and Ohta, T.(1961) Peroxidative consumption and photoreductive formation of ascorbic acid in green leaves. Plant Cell Physiol.2:31-44.
Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473-497.
Nam-Cheol, K. Kim, J.G. Lim, J.H. and Hahn, T.R. (1992). Production of secondary metabolities by tissue culture of Artemisia annua L. J. Korean Agri. Chem. Soc. 35 (2): 91-105.
Scandalios, J.G. (1993) Oxygen stress and superoxide dismutases. Plant Physiol. 101: 7-12.
Sgherri, C.L.M. Maffei, M. and Vavari-lzzo, F. (2000) Antioxidative enzymes in wheat subjected to increasing water deficit and re watering. J. Plant Physiol. 157: 273-279.
Spies, J. R.(1975) Colorimetric procedures for amino acids. In:Methods in Enzymology, Vol.3. Edited by Colowick, S.P., Kaplan, N.O. pp. 467-477. Academic Press, New York.
Sukul, N.C. Sarkar, P. Sukul, A. and Sinhababu, S.P. (1999). Antifilarial effect of Artemisia nilagirica extract and its ultra highdilutions against canine dirofilariasis, Japanese J. of Trop. Med. and Hyg. 27 (4): 477-481.
Thurman, R. G. Ley, H.G. and Scholz, R.(1972) Hepatic microsomal ethanol oxidation . Hydrogen peroxide formation and the role of catalase . Eur. J. Biochem. 25:420-430.
Published
2014-08-01