Role of Basal Media, Carbon Sources and Growth Regulators in Micropropagation of Eclipta alba – A Valuable Medicinal Herb
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Published:
Nov 12, 2018
Keywords:
axillary shoot proliferation, hardening, ex vitro, growth regulators, growth characters
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Abstract
An efficient, rapid and large-scale in vitro clonal propagation of the valuable medicinal herb Eclipta alba (Asteraceae) by enhanced auxillary shoot proliferation in cotyledonary nodal segments was designed. The medium type, various carbon sources, plant growth regulators markedly influenced in vitro propagation of E. alba. The in vitro plantlet production system was investigated on Murashige and Skoog (MS) medium with the synergistic combination of BA (4.4μM), Kin (9.2μM), 2iP (2.4 μM) and 3% sucrose which induced maximum number of shoots as well as beneficial shoot length. Subculturing of cotyledonary nodal segments on similar medium enabled continuous production of healthy shoots with similar frequency. Rooting was highest (94.3%) on full strength MS medium containing 9.8μM IBA. Micropropagated plants established in a mixture garden soil, farmyare (manure) and sand (2:1:1) were uniform and identical to the donor plant with respect to growth characteristics as well as floral features. These plants grew normally without showing any morphological variation.
Keywords: axillary shoot proliferation, hardening, ex vitro, growth regulators, growth characters
Corresponding author: E-mail: jayabalan54@yahoo.co.in, Baskaran_research@yahoo.co.in
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References
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[26] Tefera, W. and Wannakrairoj, S. 2004 Micropropagation of Krawan (Amomum krervanh Pierre ex Gagnep), Science Asia, 30, 9-15.
[27] Gao, S.L., Zhu, D.N., Cai, Z.H., Jiang, Y. and Xu, D.R. 1999 Organ Culture of a Precious Chinese Medicinal Plant-Fritillaria unibracteata. Plant Cell Tiss. Organ. Cult., 59, 97-201.
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[29] Ndoye, M., Diallo, I. and Gassama Dia, Y.K. 2003 In vitro Multiplication of the Semi-arid Forest Tree, Balanites aegyptiaca (L) Del. African Journal of Biotechnology, 2(11), 421-424.
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[32] Rajani, M.S. and URS, H.G.V.G. 1998 Multiple Shoot Induction from Cotyledonary Explants of Tabebuia spp in vitro. Phytomorphology, 48(2), 207-213.
[33] Varisai, M.S., Jawahar, M., Thiruvengadam, M., Jeyakumar, M. and Jayabalan, N. 1999 Effect of Cytokinins on the Proliferation of Multiple Shoots in Horsegram (Macrotylona uniflorum (Lam) Verdc). J. Plant Biotechnology, 1(2), 79-83.
[34] Raghava Swamy, B.V., Himabindu, K. and Lakshmi, S. 1992 In vitro Micropropagation of Elite Rosewood (Dalbergia latiflolia Roxb.). Plant Cell Rep., 11, 126-131.
[35] Thiruvengadam, M. and Jayabalan, N. 2000 Mass Propagation of Dallbergia L. in vitro. J. Plant Biotechnology, 2(3), 151-155.
[36] Sivakumar, G. and Krisnamurthy, K.V. 2000 Micropropagation of Gloriosa superba L.- an Endangered Species of Asia and Africa. Current Science, 78(1), 30-32.
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[39] Saini, R. and Jaiwal, P.K. 2000 In vitro Multiplication of Peganum harmala an Important Medicinal Plant. Indian J. Exp. Biol., 38, 499-503.
[40] Martin, K.P. 2000 Rapid Propagation of Holostemma ada-kodien Schult., A Rare Medicinal Plant, through Axillary Bud Multiplication and Indirect Organogenesis. Plant Cell Rep., 21, 112-117.
[41] Marks, T.R. and Simpson, S.E. 1994 Factors Affecting Shoot Development in Apically Dominant Acer Cultivars in vitro. J. Hort. Sci., 69, 543-551.
[42] Preece, J.E,. Huttreman, C.A., Ashby, W.C. and Roth, P.L. 1991 Micro Cutting Propagation of Silver Maple I. Results with Adult and Juvenile Propagules. J. Am. Soc. Hort. Sci., 116, 142-148.
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[44] Thompson, M. and Thorpe, T. 1987 Metabolic and Non-metabolic Roles of Carbohydrates. In: Bonga, J.M. and Durzan, D.J., Ed. Cell and Tissue Culture in Forestry. Martinus Nijhoff Publishers, Dordrecht, pp. 89-112.
[45] Romano, A., Noronha, C. and Martins-Loucao, M.A. 1995 Role of Carbohydrates in Micropropagation of Cork Oak. Plant Cell Tiss. Org. Cult., 40, 159-167.
[46] Fatima, S., Sandeep, K. and Yogeshwar, M. 2000 In vitro Plantlet Production System for Kaempferi galanga, A Rare Indian Medicinal Herb. Plant Cell Tiss. Org. Cult., 63, 193-197.
[47] Fuentes, S.R.L., Calheiros, M.B.P., Manetti-Filho, J. and Vieira, L.G.E. 2000 The Effects of Silver Nitrate and Different Carbohydrate Sources on Somatic Embryogenesis in Coffea canephora. Plant Cell Tiss. Org. Cult., 60, 5-13.
[48] Borkowska, B. and 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.
[49] Hisashi, H. and Yasuhiro, M. 1996 Micropropagation of Prunus Mume. Plant Cell Tiss. Organ. Cult., 46, 265-267.
[50] Marino, G., Magnanini, E., Battistini, S. and Righetti, B. 1991 Effect of Hormones and Main Carbon Energy Sources on in vitro Propagation of Apricot (Prunus armeniaca L.) cvs. ‘San Castrese’ and ‘Portici’. Acta Hort., 293, 355-362.
[51] de Paiva Neto, V.B., da Mota, T.R. and Otani, W.C. 2003 Direct Organogenesis from Hypocotyl-derived Explants of Annatto (Bixa Orellana). Plant Cell Tiss. Org. Cult., 75, 159-167.
[52] Pua, E.C. and Chong, C. 1984 Requirement for Sorbitol (D-glucitol) as Carbon Source for in vitro Propagation of Malus robusta No.5. Canadian Journal of Botany, 62, 1545-1549.
[53] Pritchard, J., Wyn-Jones, R.G. and Tomos, A.D. 1991 Turgor, Growth and Rheological Gradients in Wheat Roots following Osmotic, Stress. J. Exp. Bot., 42, 1043-1049.
[54] Venkatachalam, P. and Jayabalan, N. 1997 Effect of Auxins and Cytokinins on Efficient Plant Regeneration and Multiple Shoot Formation from Cotyledons and Cotyledonary Node Explants of Ground Nut (Arachis hypogaea L.) by in vitro Culture Technology. Applied Biochemistry and Biotechnology, 67, 237-247.
[55] Purohit, S.D. and Ashish, D. 1984 Micropropagation of Sterculia urens Roxb.-an Endangered Tree Species. Plant Cell Rep., 15, 704-706.
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[2] Murphy, R.C., Hammarstrom, S., Samuelsson, B. and Leukotriene, C. 1979 A Slow Reacting Substance from Marine Mastocytoma Cells. Proc. Natl. Acad. Sci., USA, pp 4275-4279.
[3] Orning, L., Hammarstrom, S., Samuelsson, B. and Leukotriene, D. 1980 A Slow Reacting Substance from Rat Basophilic Leukemia Cells. Proc. Natl. Acad. Sci., USA. pp.2014-2017.
[4] Wagner, H., Geyer, B., Kiso, Y. and Rao, G.S. 1986 Coumestans as the Main Active Principles of the Liver Drugs Eclipta alba and Wedelia calendulacea. Planta Med., 52, 370-373
[5] Franca, S.C., Bertoni, B.W., and Pereira, A.M.S. 1995 Antihepatotoxic Agent in Micropropagated Plantlets of Eclipta alba. Plant Cell Tiss. Organ. Cult., 40, 297-299.
[6] Mors, W.B., Nascimento, M.C., Parente, J.P., Silva, M.H., Melo, P.A. and Suarez-Kurtz, G. 1989 Neutralization of Lethal and Myotoxic Activities of South American Rattlesnakes Venom by Extracts and Constituents of the Plant Eclipta prostate (Asteraceae). Toxicon., 27, 1003-1009.
[7] Syed, S.D., Deepak, M., Yogisha, S., Chandrashekar, A.P., Muddarachappa, K.A., D’Souza, P., Agarwal, A. and Venkataraman, B.V. 2003 Trypsin Inhibitory Effect of Wedelolactone and Demethylwedelolactone. Phytother. Res.,17(4), 420-421.
[8] Upadhyay, R.K., Pandey, M.B., Jha, R.N. and Pandey, V.B. 2001 Eclalbatin, a Triterpene Saponin from Eclipta alba. J. Asian Nat. Prod. Res., 3(3), 213-217.
[9] Sawant, M., Isaac, J.C. and Narayanan, S. 2004 Analgesic Sudies on total Alkaloids and Alcohol Extracts of Eclipta alba (Linn.) Hassk. Phytother. Res., 18(2), 111-113.
[10] Sen, J. and Sharma, A.K. 1991 Micropropagation of Withania somnifera from Germinating Seeds and Shoot Tips. Plant Cell Tiss. Org. Cult., 26, 71-73.
[11] Pandey, N.K., Tewari, K.C., Tewari, R.N., Joshi, G.C., Pande, V.N. and Pandey, G. 1993 Medicinal Plants of Kumaon Himalaya, Strategies for Conservation. In: Dhar, U., Ed. Himalayan Biodiversity Conservation Strategies. Himavikas publication, pp. 293-302.
[12] Sing, 1998 Himalayan Herbs and Drugs, Importance and Extinction Threat. J.Sci. Res. Plants Med., 10, 47-52.
[13] Sharma, B.L. and Kumar, A. 1998 Biodiversity of Medicinal Plant of Triyugi Narain (Garhwal Himalaya) and their Conservation. National Conference on Recent Trends in Spices & Medicinal Plant Research, Calcutta, WB, India, A-78.
[14] Ajithkumar, D. and Seeni, S. 1998 Rapid Clonal Multiplication through in vitro Axillary Shoot Proliferation of Aegle marmelos (L.) Corr., A Medicinal Tree. Plant Cell Rep., 17, 422-426.
[15] Sahoo, Y. and Chand, P.K. 1998 Micropropagation of Vitex negundo L., a Woody Aromatic Medicinal Shrub, Through High Frequency Axillary Shoot Proliferation. Plant Cell Rep., 18, 301-307.
[16] Prakash, E., Sha Valli Khan, P.S., Sairam Reddy. P. and Rao, K.R. 1999 Regeneration of Plants from Seed-derived Callus of Hybanthus enneaspermus L. Muell., a Rare Ethnobotanical Herb. Plant Cell Rep., 18, 873-878.
[17] Tejavathi, D.H. and Shailaja, K.s. 1999 Regeneration of Plants from the Cultures of Bacopa monnieri (L.) Pennell. Phytomorphology, 49(4), 447-452.
[18] Borthakur, M., Dutta, K., Nath, S.C. and Sing, R.S. 2000 Micropropagation of Eclipta alba and Eupatorium adenophorum using a Single-step Nodal Cutting Technigue. Plant Cell Tiss. Organ. Cult., 62, 239-242.
[19] Murashige, T. and Skoog, F. 1962 A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Culture. Physiol. Plant, 15, 473-479.
[20] Gamborg, O.L., Miller, R.A. and Ojima, K. 1968 Nutrient Requirements of Suspension Cultures of Soybean Root Cells. Exp. Cell Res., 50, 151-158.
[21] Schenk, R.U. and Hildebrandt, A.C. 1972 Medium and Technique for Induction and Growth of Monocotyledonous and Dicotyledonous Plant Cell Cultures. Can. J. Bot., 50, 199-204.
[22] Gomez, K.A. and Gomez, A.A. 1976 Statistical Procedures for Agricultural Research with Emphasis on Rice, Los Banos, Philippines, International Rice Research Institute.
[23] Shekhawat, N.S., Rathore, T.S., Singh, R.P., Deora, N.S. and Rao, S.R. 1993 Factors Affecting In vitro Clonal Propagation of Prosopid vinrtis. Plant Growth Reg., 12, 273-280.
[24] Das, S., Timir, B.J. and Sumita, J. 1996 In vitro Propagation of Cashewnut. Plant Cell Rep., 15, 615-619.
[25] Ramage, C.M. and Williams, R.R. 2002 Mineral Nutrition and Plant Morphogenesis. In Vitro Cell Dev. Biol-Plant, 38, 116-124.
[26] Tefera, W. and Wannakrairoj, S. 2004 Micropropagation of Krawan (Amomum krervanh Pierre ex Gagnep), Science Asia, 30, 9-15.
[27] Gao, S.L., Zhu, D.N., Cai, Z.H., Jiang, Y. and Xu, D.R. 1999 Organ Culture of a Precious Chinese Medicinal Plant-Fritillaria unibracteata. Plant Cell Tiss. Organ. Cult., 59, 97-201.
[28] Komalavalli, N. and Rao, M.V. 2000 In vitro Micropropagation of Gymnema sylvestre-A Multipurpose Medicinal Plant. Plant Cell Tiss. Org. Cult., 61, 97-105.
[29] Ndoye, M., Diallo, I. and Gassama Dia, Y.K. 2003 In vitro Multiplication of the Semi-arid Forest Tree, Balanites aegyptiaca (L) Del. African Journal of Biotechnology, 2(11), 421-424.
[30] Bajaj, Y.P.S., Reghunath, B.R. and Gopalakrishnan, P.K. 1993 Elettaria cardamomum Maton(cardamom): Aromatic Compounds, in vitro Culture Studies and Clonal Propagation. In: Biotechnology in Agriculture and Forestry, Medicinal and Aromatic Plants IV, Springer- Verlag Berlin Heidelberg. Germany pp.132-47.
[31] Indra, D. and Uppeandra, D. 2000 Micropropagation of Indian Wild Strawberry. Plant Cell Tiss. Org. Cult., 60, 83-88.
[32] Rajani, M.S. and URS, H.G.V.G. 1998 Multiple Shoot Induction from Cotyledonary Explants of Tabebuia spp in vitro. Phytomorphology, 48(2), 207-213.
[33] Varisai, M.S., Jawahar, M., Thiruvengadam, M., Jeyakumar, M. and Jayabalan, N. 1999 Effect of Cytokinins on the Proliferation of Multiple Shoots in Horsegram (Macrotylona uniflorum (Lam) Verdc). J. Plant Biotechnology, 1(2), 79-83.
[34] Raghava Swamy, B.V., Himabindu, K. and Lakshmi, S. 1992 In vitro Micropropagation of Elite Rosewood (Dalbergia latiflolia Roxb.). Plant Cell Rep., 11, 126-131.
[35] Thiruvengadam, M. and Jayabalan, N. 2000 Mass Propagation of Dallbergia L. in vitro. J. Plant Biotechnology, 2(3), 151-155.
[36] Sivakumar, G. and Krisnamurthy, K.V. 2000 Micropropagation of Gloriosa superba L.- an Endangered Species of Asia and Africa. Current Science, 78(1), 30-32.
[37] Hu, C.Y. and Wang, P.J. 1983 Meristem Shoot Tip and Bud Culture. In: Evans, D.A., Sharp, W.R., Ammirato, P.V. and Yamada, Y., Ed. Hand Book of Plant Cell Culture. Macmillan, New York, pp. 177-227.
[38] Gulati, A. and Jaiwal, P.K. 1994 Plant Regeneration from Gotyledonary Node Explants of Mung Bean (Vigna radiata). Plant Cell Rep., 13, 523-527.
[39] Saini, R. and Jaiwal, P.K. 2000 In vitro Multiplication of Peganum harmala an Important Medicinal Plant. Indian J. Exp. Biol., 38, 499-503.
[40] Martin, K.P. 2000 Rapid Propagation of Holostemma ada-kodien Schult., A Rare Medicinal Plant, through Axillary Bud Multiplication and Indirect Organogenesis. Plant Cell Rep., 21, 112-117.
[41] Marks, T.R. and Simpson, S.E. 1994 Factors Affecting Shoot Development in Apically Dominant Acer Cultivars in vitro. J. Hort. Sci., 69, 543-551.
[42] Preece, J.E,. Huttreman, C.A., Ashby, W.C. and Roth, P.L. 1991 Micro Cutting Propagation of Silver Maple I. Results with Adult and Juvenile Propagules. J. Am. Soc. Hort. Sci., 116, 142-148.
[43] Kozai, T. 1991 Micropropagation under Photoautotrophic Conditions. In: Debergh, P.C., Zimmerman, R.H., Ed. Micropropagation: Technology and Application. Kluwer Academic Publishers, Dordrecht, pp. 447-469.
[44] Thompson, M. and Thorpe, T. 1987 Metabolic and Non-metabolic Roles of Carbohydrates. In: Bonga, J.M. and Durzan, D.J., Ed. Cell and Tissue Culture in Forestry. Martinus Nijhoff Publishers, Dordrecht, pp. 89-112.
[45] Romano, A., Noronha, C. and Martins-Loucao, M.A. 1995 Role of Carbohydrates in Micropropagation of Cork Oak. Plant Cell Tiss. Org. Cult., 40, 159-167.
[46] Fatima, S., Sandeep, K. and Yogeshwar, M. 2000 In vitro Plantlet Production System for Kaempferi galanga, A Rare Indian Medicinal Herb. Plant Cell Tiss. Org. Cult., 63, 193-197.
[47] Fuentes, S.R.L., Calheiros, M.B.P., Manetti-Filho, J. and Vieira, L.G.E. 2000 The Effects of Silver Nitrate and Different Carbohydrate Sources on Somatic Embryogenesis in Coffea canephora. Plant Cell Tiss. Org. Cult., 60, 5-13.
[48] Borkowska, B. and 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.
[49] Hisashi, H. and Yasuhiro, M. 1996 Micropropagation of Prunus Mume. Plant Cell Tiss. Organ. Cult., 46, 265-267.
[50] Marino, G., Magnanini, E., Battistini, S. and Righetti, B. 1991 Effect of Hormones and Main Carbon Energy Sources on in vitro Propagation of Apricot (Prunus armeniaca L.) cvs. ‘San Castrese’ and ‘Portici’. Acta Hort., 293, 355-362.
[51] de Paiva Neto, V.B., da Mota, T.R. and Otani, W.C. 2003 Direct Organogenesis from Hypocotyl-derived Explants of Annatto (Bixa Orellana). Plant Cell Tiss. Org. Cult., 75, 159-167.
[52] Pua, E.C. and Chong, C. 1984 Requirement for Sorbitol (D-glucitol) as Carbon Source for in vitro Propagation of Malus robusta No.5. Canadian Journal of Botany, 62, 1545-1549.
[53] Pritchard, J., Wyn-Jones, R.G. and Tomos, A.D. 1991 Turgor, Growth and Rheological Gradients in Wheat Roots following Osmotic, Stress. J. Exp. Bot., 42, 1043-1049.
[54] Venkatachalam, P. and Jayabalan, N. 1997 Effect of Auxins and Cytokinins on Efficient Plant Regeneration and Multiple Shoot Formation from Cotyledons and Cotyledonary Node Explants of Ground Nut (Arachis hypogaea L.) by in vitro Culture Technology. Applied Biochemistry and Biotechnology, 67, 237-247.
[55] Purohit, S.D. and Ashish, D. 1984 Micropropagation of Sterculia urens Roxb.-an Endangered Tree Species. Plant Cell Rep., 15, 704-706.
[56] Constantine, D. 1978 Round Table Conference, Gembloux, Belgium. p 134