Back
 AJPS  Vol.6 No.19 , December 2015
Effect of Nutrient Media and KNO3 on in Vitro Plant Regeneration in Saraca asoca (Roxb.) Willd
Abstract: Forest trees in general and those belonging to family Fabaceae in particular, have proved to be recalcitrant for propagation through tissue culture. Saraca asoca (Roxb.) Willd. (Family-Caesalpinaceae) is one such tree which has become vulnerable in nature due to over exploitation of its bark. Four nutrient media [MS (Murashige and Skoog Medium), WPM (Woody Plant Medium), B5 (Gamborg’s Medium) and NN (Nitsch and Nitsch Medium, 1969)] and five doses of BA (N6-Benzyladenine) (0, 2.2, 4.4, 8.8 and 17.8 μM) and their all possible interactions were tested for shoot induction and proliferation from nodal segments of 3-year-old plants. B5 medium supplemented with 2.2 μM BA was screened out as the most suitable medium shoot induction, proliferation and elongation of regenerated shoots. In order to enhance shoot number, the nitrogen source in B5 medium was modified and five strengths of KNO3 (0.25×, 0.5×, 1.0×, 1.25× and 1.5×) were tested. The different strengths of KNO3 (Potassium nitrate) had statistically significant effect on number of shoots and on 0.25× strength of KNO3, maximum number of shoots (1.92) were obtained. The modified strengths of KNO3 did not significantly affect the elongation of shoots. Effect of 5 durations (quick dip, transfer of shoots after 1 day, after 3 days, after 5 days and after 7 days) of pulse treatment with 200 μM IBA (Indole-3-butyric acid) in 1/2 strength MS liquid medium was tested. Thereafter, the shoots were transferred to semi-solid half strength MS medium supplemented with 0.2 μM IBA and 3.96 μM phloroglucinol. Pulse treatment of 5 days duration resulted in 37.5% in vitro rooting of shoots. Plantlets were hardened in soilrite soaked with half strength MS medium in culture room and later shifted to a soil mixture in shade house.
Cite this paper: Shirin, F. , Parihar, N. and Shah, S. (2015) Effect of Nutrient Media and KNO3 on in Vitro Plant Regeneration in Saraca asoca (Roxb.) Willd. American Journal of Plant Sciences, 6, 3282-3292. doi: 10.4236/ajps.2015.619320.
References

[1]   Troup, R.S. (1983) The Silviculture of Indian Trees: Leguminosae (Caesalpinieae) to Verbenaceae. Vol. 4, The Controller of Publications, Government of India, Delhi.

[2]   Noorunnisa Begum, S., Ravikumar, K. and Ved, D.K. (2014) “Asoka”—An Important Medicinal Plant, Its Market Scenario and Conservation Measures in India. Current Science, 107, 26-28.

[3]   Kirtikar, K.R. and Basu, B.D. (1981) Indian Medicinal Plants. Vol. 2, International Book Distribution, Dehradun.

[4]   Garg, S., Chandra, A., Mazumder, A. and Mazumder, R. (2014) Analgesic Potential of Hydrogels of Silver Nanoparticles Using Aqueous Extract of Saraca indica Bark. International Journal of Pharmaceutical Science Research, 5, 240-245.

[5]   Anon (1985) The Wealth of India, Raw Materials. Vol. 4, CSIR, New Delhi, 232-234.

[6]   IUCN (2010) IUCN Red List of Threatened Species. Version 2010.
www.iucnredlist.org

[7]   Parkash, R., Choudhary, D.C. and Negi, S.S. (1991) Propagation Practices of Important Indian Trees. International Book Distributors, Dehradun.

[8]   Rama Subbu, R., Chandraprabha, A. and Sevuga, P. (2008) In Vitro Clonal Propagation of Vulnerable Medicinal Plant Saraca asoca (Roxb.) De Wilde. Natural Product Radiance, 7, 338-341.

[9]   Shirin, F., Parihar, N., Rana, P.K. and Ansari, S.A. (2011) In Vitro Shoot Regeneration from Embryonic Axis of a Multipurpose Vulnerable Leguminous Tree, Saraca indica L. Tree and Forestry Science and Biotechnology, 5, 45-48.

[10]   Murashige, T. and Skoog, F. (1962) A Revised Medium for Rapid Growth and Bioassay with Tobacco Tissue Culture. Physiologia Plantarum, 15, 473-497.
http://dx.doi.org/10.1111/j.1399-3054.1962.tb08052.x

[11]   Gamborg, O.L., Miller, R.A. and Ojima, K. (1968) Nutrient Requirements of Suspension Cultures of Soyabean Root Cells. Experimental Cell Research, 50, 151-158.
http://dx.doi.org/10.1016/0014-4827(68)90403-5

[12]   Lloyd, G. and McCown, B.H. (1980) Commercially-Feasible Micropropagation of Mountain Laurel, Kalmia latifolia, by Use of Shoot-Tip Culture. Combined Proceedings-International Plant Propagator’s Society, 30, 421-427.

[13]   Nitsch, J.P. and Nitsch, C. (1969) Haploid Plants from Pollen Grains. Science, 163, 85-87.
http://dx.doi.org/10.1126/science.163.3862.85

[14]   Gomez, K.A. and Gomez, A.A. (1984) Statistical Procedures for Agricultural Research. 2nd Edition, John Willey and Sons, New York.

[15]   Gamborg, O.L. and Phillips, G.C. (1995) Media Preparation and Handling. In: Gamborg, O.L. and Phillips, G.C., Eds., Plant Cell, Tissue and Organ Culture—Fundamental Methods, Springer-Verlag, Berlin, 21-34.
http://dx.doi.org/10.1007/978-3-642-79048-5_2

[16]   Smith, R.H. (2000) Plant Tissue Culture Techniques and Experiments. Academic Press, Waltham.

[17]   Rugini, E. (1984) In Vitro Propagation of Some Olive (Olea europaea sativa L.) Cultivars with Different Root-Ability, and Medium Development Using Analytical Data from Developing Shoots and Embryos. Scientia Hotriculturae, 24, 123-134.
http://dx.doi.org/10.1016/0304-4238(84)90143-2

[18]   Mehta, U.J., Krishnamurthy, K.V. and Hazra, S. (2000) Regeneration of Plants via Adventitious Bud Formation from Mature Zygotic Embryo Axis of Tamarind (Tamarindus indica L.). Current Science, 78, 1231-1234.

[19]   Lu, M.C. (2005) Micropropagation of Vitis thunbergii Sieb. et Zucc., a Medicinal Herb, through High-Frequency Shoot Tip Culture. Scientia Hotriculturae, 107, 64-69.
http://dx.doi.org/10.1016/j.scienta.2005.05.014

[20]   Jain, N., Bairu, M.W., Stirk, W.A. and Van Staden, J. (2009) The Effect of Medium, Carbon Source and Explant on Regeneration and Control of Shoot-Tip Necrosis in Harpagophytum procumbens. South African Journal of Botany, 75, 117-121.
http://dx.doi.org/10.1016/j.sajb.2008.08.005

[21]   Vlachova, M., Metz, B.A., Schell, J. and de Bruijn, F.J. (1987) The Tropical Legume Sesbania rostrata: Tissue Culture, Plant Regeneration and Infection with Agrobacterium tumefaciens and Rhizogenes Strains. Plant Science, 50, 213-223.
http://dx.doi.org/10.1016/0168-9452(87)90076-8

[22]   Brandt, E.B. and Hess, D. (1994) In Vitro Regeneration and Propagation of Chick Pea (Cicer arietinum L.) from Meristem Tips and Cotyledonary Nodes. In Vitro Cellular and Developmental Biology-Plant, 30, 75-80.
http://dx.doi.org/10.1007/BF02632124

[23]   Diallo, M.S., Ndiaye, A., Sagna, M. and Gassama-Dia, Y.K. (2008) Plants Regeneration from African Cowpea Variety (Vigna unguiculata L. Walp.). African Journal of Biotechnology, 7, 2828-2833.

[24]   Berger, K. and Schaffner, W. (1995) In Vitro Propagation of the Leguminous Tree Swartzia madagascariensis. Plant Cell, Tissue and Organ Culture, 40, 289-291.
http://dx.doi.org/10.1007/BF00048136

[25]   Ye, G., Mcneil, D.L., Conner, A.J. and Hill, G.D. (2002) Multiple Shoot Formation in Lentil (Lens culinaris) Seeds. New Zealand Journal of Crop and Horticultural Science, 30, 1-8.
http://dx.doi.org/10.1080/01140671.2002.9514193

[26]   Suri, S.S., Jain, S. and Ramawat, K.G. (1999) Plantlet Regeneration and Bulbil Formation in Vitro from Leaf and Stem Explants of Curculigo orchioides, an Endangered Medicinal Plant. Scientia Hotriculturae, 79,127-134.
http://dx.doi.org/10.1016/S0304-4238(98)00118-6

[27]   Burnouf-Radosevich, M. and Paupardin, C. (1985) Vegetative Propagation of Chenopodium quinoa by Shoot Tip Culture. American Journal of Botany, 72, 278-283.
http://dx.doi.org/10.2307/2443555

[28]   Tabone, T.J., Felker, P., Bungham, R.L., Reyes, I. and Loughery, S. (1986) Techniques in the Shoot Multiplication of the Leguminous Tree Prosopis alba Clone B2V5o. Forest Ecology and Management, 16, 191-200.
http://dx.doi.org/10.1016/0378-1127(86)90019-8

[29]   Shrivastava, S. and Chawla, H.S. (2001) Synergistic Effect of Growth Regulators and Glutamine on Regeneration Response in High Yielding Cultivars of Wheat (Triticum aestivum L.). Indian Journal of Genetics and Plant Breeding, 61, 12-15.

[30]   Zamir, R., Ali, N., Shah, S.T., Muhammad, T. and Shah, S.A. (2007) In Vitro Re-Generation of Guava (Psidium guajava L.) from Shoot Tips of Mature Trees. Pakistan Journal of Botany, 39, 2395-2398.

[31]   Patil, G., Deokar, A., Jain, P.K., Thengane, R.J. and Srinivasan, R. (2009) Development of a Phosphomannose Isomerase-Based Agrobacterium-Mediated Transformation System for Chickpea (Cicer arietinum L.). Plant Cell Reports, 28, 1669-1676.
http://dx.doi.org/10.1007/s00299-009-0766-3

[32]   De Neto, V.B.P. and Otoni, W.C. (2003) Carbon Sources and Their Osmotic Potential in Plant Tissue Culture: Does It Matter? Scientia Hotriculturae, 97, 193-202.
http://dx.doi.org/10.1016/S0304-4238(02)00231-5

[33]   Thorpe, T.A. (1982) Carbohydrate Utilization and Metabolism. In: Bonga, J.M. and Durzan, D.J., Eds., Tissue Culture in Forestry, Martinus Nijhoff/Dr. W. Junk Publishers, The Hague, 325-368.
http://dx.doi.org/10.1007/978-94-017-3538-4_11

[34]   Lemos, E.E.P. and Baker, D.A. (1995) Shoot Regeneration in Response to Carbon Source on Internodal Explants of Annona muricata L. Plant Growth Regulation, 25, 106-112.

[35]   Ahmad, T., Abbasi, N.A., Ahmad, H.I. and Ali, A. (2007) Comparison of Sucrose and Sorbitol as Main Carbon Energy Sources in Micropropagation of Peach Rootstock GF-677. Pakistan Journal of Botany, 39, 1269-1275.

[36]   Kabir, A.H., Bari, M.A., Huda, A.K.M.N., Rezvy, M.A. and Mahfuz, I. (2007) Effect of Growth Regulators and Carbon Sources on Axillary Shoot Proliferation from Shoot-Tip Explant and Successful Transplantation of Papaya (Carica papaya L.). Biotechnology, l6, 268-272.

[37]   Shanjani, P.S. (2003) Nitrogen Effect on Callus Induction and Plant Regeneration of Juniperus excelsa. International Journal of Agriculture and Biological Engineering, 5, 419-422.

[38]   Vinterhalter, B., Ninkovic, S., Zdravkovic-Korac, S., Subotic, A. and Vinterhalter, D. (2007) Effect of Nitrogen Salts on the Growth of Ceratonia siliqua L. Shoot Cultures. Archives of Biological Science, 59, 217-222.
http://dx.doi.org/10.2298/ABS0703217V

[39]   Shekhawat, N.S., Rathore, T.S., Singh, R.P., Deora, N.S. and Rao, S.R. (1993) Factors Affecting in Vitro Clonal Propagation of Prosopis cineraria. Plant Growth Regulation, 12, 273-280.
http://dx.doi.org/10.1007/BF00027208

[40]   Gaspar, T. and Coumans, M. (1987) Root Formation. In: Bonga, J.M. and Durzan, D.J., Eds., Cell and Tissue Culture in Forestry, Vol. 2, Martinus Nijhoff, Dordrecht, 202-217.
http://dx.doi.org/10.1007/978-94-009-4484-8_10

[41]   Zaerr, J.B. and Mapes, M.O. (1982) Action of Growth Regulators. In: Bonga, J.M. and Durzan, D.J., Eds., Tissue Culture in Forestry, Martinus Nijhoff/Dr. W. Junk Publishers, The Hague, 231-255.
http://dx.doi.org/10.1007/978-94-017-3538-4_9

[42]   Epstein, E. and Ludwig-Muller, J. (1993) Indole-3-Butyric Acid in Plants: Occurrence, Synthesis, Metabolism and Transport. Physiologia Plantarum, 88, 382-389.
http://dx.doi.org/10.1111/j.1399-3054.1993.tb05513.x

[43]   Martin, K.P. (2002) Rapid Propagation of Holostemma ada-kodein Schult., a Rare Medicinal Plant, through Axillary Bud Multiplication and Indirect Organogenesis. Plant Cell Reports, 21, 112-117.
http://dx.doi.org/10.1007/s00299-002-0483-7

[44]   Devi, Y.S., Mukherjee, B.B. and Gupta, S. (1994) Rapid Cloning of Elite Teak (Tectona grandis Linn.) by in Vitro Multiple Shoot Production. Indian Journal of Experimental Biology, 32, 668-671.

[45]   Tiwari, S.K., Tiwari, K.P. and Siril, E.A. (2002) An Improved Micropropagation Protocol for Teak. Plant Cell Tissue and Organ Culture, 71, 1-6.
http://dx.doi.org/10.1023/A:1016570000846

[46]   Shelby, C., Kennedy, J. and Harvey, M.R. (1992) Adventitious Root Formation in Hypocotyl Cuttings of Picea sitchensis (Bong.) Carr.—The Influence of Plant Growth Regulators. New Phytologist, 120, 453-457.
http://dx.doi.org/10.1111/j.1469-8137.1992.tb01792.x

[47]   Purohit, S.D. and Kudka, G. (2004) Micropropagation of an Adult Tree—Wrightia tinctoria. Indian Journal of Biotechnology, 3, 216-220.

[48]   Chabukswar, S. and Deodhar, M. (2005) Rooting and Hardening of in Vitro Plantlets of Garcinia indica Chois. Indian Journal of Biotechnology, 4, 409-413.

[49]   Husain, M.K., Anis, M. and Shahzad, A. (2007) In Vitro Propagation of Indian Kino (Pterocarpus marsupium Roxb.) Using Thidiazuron. In Vitro Cellular and Developmental Biology-Plant, 43, 59-64.
http://dx.doi.org/10.1007/s11627-006-9011-8

[50]   Shasthree, T., Imran, M.A. and Mallaiah, B. (2009) In Vitro Rooting from Callus Cultures Derived from Seedling Explants of Erythrina variegate L. Current Trends in Biotechnology and Pharmacy, 4, 447-452.

[51]   De Klerk, G.J., Vander Kreiken, W. and de Jong, J.C. (1999) The Formation of Adventitious Roots: New Concepts, New Possibilities. In Vitro Cellular and Developmental Biology-Plant, 35,189-199.
http://dx.doi.org/10.1007/s11627-999-0076-z

[52]   James, D.J. (1983) Adventitious Root Formation in Vitro in Apple Rootstocks (Malus pumila) I. Factors Affecting the Length of the Auxin-Sensitive Phase in M.9. Physiologia Plantarum, 57, 149-153.
http://dx.doi.org/10.1111/j.1399-3054.1983.tb00745.x

[53]   Zanol, G.C., de Fortes, G.R.L., Campos, A.D., Centellas, A.Q. and da Silva, J.B. (1998) In Vitro Rooting and Peroxidase Activity of Apple Rootstock cv, Marubakadio Treated with Indole Butyric Acid and Phloroglucinol. Revista Brasileira de Fisiologia Vegetal, 10, 65-68.

[54]   Hammatt, N. and Grant, N.J. (1997) Micropropagation of Mature British Wild Cherry. Plant Cell, Tissue and Organ Culture, 47, 103-110.
http://dx.doi.org/10.1007/BF02318945

[55]   Bhojwani, S.S. and Razdan, M.K. (1996) Plant Tissue Culture: Theory and Practice. Elsevier Science, Amsterdam.

 
 
Top