biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 55:164-168, 2011 | DOI: 10.1007/s10535-011-0023-9

In vitro cloning of Azadirachta indica from root explants

K. Arora1, M. Sharma1, J. Srivastava2, S. A. Ranade2, A. K. Sharma1,*
1 Tissue Culture Laboratory, National Botanical Research Institute, Lucknow, India
2 Plant Molecular Biology (Genomics) Division, National Botanical Research Institute, Lucknow, India

In vitro cultures of Azadirachta indica A. Juss. were raised by first culturing the root segments on modified Murashige and Skoog (MS) medium supplemented with 8.88 μM 6-benzylaminopurine (BAP), 9.84 μM N6-(2-isopentenyl) adenine (2iP), 5.71 μM indole-3-acetic acid (IAA), 81.43 μM adenine hemisulphate and 2.27 μM putrescine for 2 d followed by their transfer to the same medium except containing one-tenth of the initially used concentrations of BAP, 2iP and IAA. The regenerated shoots sustained proliferation in the basal medium supplemented with 1.11 μM BAP, 1.43 μM IAA and 135.72 μM adenine hemisulphate. The isolated shoots were rooted to produce plantlets in the presence of 2.46 μM indole-3-butyric acid (IBA). The plantlets showed uniform luxuriant growth under field conditions. True-to-type nature of the field-grown root-regenerated plants was ascertained by random amplified polymorphic DNA (RAPD) analysis.

Keywords: auxins; cytokinins; genetic stability; neem; RAPD; root-regenerated plants

Received: April 15, 2009; Accepted: January 15, 2010; Published: March 1, 2011  Show citation

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Arora, K., Sharma, M., Srivastava, J., Ranade, S.A., & Sharma, A.K. (2011). In vitro cloning of Azadirachta indica from root explants. Biologia plantarum, 55(1), 164-168. doi: 10.1007/s10535-011-0023-9
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    References

    1. Arora, K., Sharma, M., Sharma, A.K.: Control of pattern of regenerant differentiation and plantlet production from leaflet segments of Azadirachta indica A. Juss. (neem). - Acta Physiol. Plant. 31: 371-378, 2009. Go to original source...
    2. Arora, K., Sharma, M., Srivastava, J., Ranade, S.A., Sharma A.K.: Rapid in vitro cloning of a 40-year-old tree of Azadirachta indica A. Juss. (Neem) employing nodal stem segments. - Agroforest. Syst. 78: 53-63, 2010. Go to original source...
    3. Bais, H.P., Sudha, G.S., Ravishankar, G.A.: Putrescine and silver nitrate influences shoot multiplication, in vitro flowering and endogenous titers of polyamines in Cichorium intybus L. cv. Lucknow local. - J. Plant Growth Regul. 19: 238-248, 2000. Go to original source...
    4. Chandrika, M., Thoyajaksha, Ravishankar Rai, V., Ramachandra Kini, K.: Assessment of genetic stability of in vitro grown Dictyospermum ovalifolium. - Biol. Plant. 52: 735-739, 2008. Go to original source...
    5. Chaturvedi, H.C., Sharma, M., Sharma, A.K., Jain, M., Agha, B.Q., Gupta, P.: In vitro germplasm preservation through regenerative excised root culture for conservation of phytodiversity. - Indian J. Biotechnol. 3: 305-315, 2004a.
    6. Chaturvedi, R., Razdan, M.K., Bhojwani, S.S.: In vitro clonal propagation of an adult tree of neem (Azadirachta indica A. Juss.) by forced axillary branching. - Plant Sci. 166: 501-506, 2004b. Go to original source...
    7. Chi, G.L., Lin, W.S., Lee, JEE., Pua, E.C.: Role of polyamines on de novo shoot morphogenesis from cotyledons of Brassica campestris spp. Pekinensis (Lour) Olsson in vitro. - Plant Cell Rep. 13: 323-329, 1994. Go to original source...
    8. De Kochko, A., Hamon, S.: A rapid and efficient method for the isolation of restrictable total DNA from plants of genus Abelmoschus. - Plant mol. Biol. Rep. 8: 3-7, 1990. Go to original source...
    9. D'Onofrio, C., Morini, S.: Simultaneous regeneration of different morphogenic structures from quince leaves as affected by growth regulator combination and treatment length. - Biol. Plant. 47: 321-325, 2003/2004. Go to original source...
    10. Dogra, P.D., Thapliyal, R.C.: Gene resources and breeding potential. - In: Randhawa, N.S., Parmar, B.S. (ed.): Neem. Pp. 27-32. New Age International, New Delhi 1996.
    11. Eeswara, J.P., Stuchbury, T., Allan, E.J., Mordue (Luntz), A.J.: A standard procedure for the micropropagation of the neem tree (Azadirachta indica A. Juss.). - Plant Cell Rep. 17: 215-219, 1998. Go to original source...
    12. Ezumah, B.S.: Germination and storage of neem (Azadirachta indica A. Juss.) seed. - Seed Sci. Technol. 14: 593-600, 1986.
    13. Farooqui, N., Ranade, S.A., Sane, P.V.: RAPD profile variation amongst provenances of neem. - Biochem. mol. Biol. Int. 45: 931-939, 1998. Go to original source...
    14. George, E.F.: Plant Propagation by Tissue Culture. Part I The Technology. - Exegetics, Edington 1993.
    15. Hu, Z., Hu, Y., Gao, H.H., Guan, X.Q., Zhuang, D.H.: Callus production, somatic embryogenesis and plant regeneration of Lycium barbarum root explants. - Biol. Plant. 52: 93-96, 2008. Go to original source...
    16. Huang, W.J., Ning, G.G., Liu, G.F., Bao, M.Z.: Determination of genetic stability of long-term micropropagated plantlets of Platanus acerifolia using ISSR markers. - Biol. Plant. 53: 159-163, 2009. Go to original source...
    17. Hussain, Z., Tyagi, R.K., Sharma, R., Agrawal, A.: Genetic diversity in in vitro-conserved germplasm of Curcuma L. as revealed by RAPD markers. - Biol. Plant. 52: 627-633, 2008. Go to original source...
    18. Johansen, D.A.: Plant Microtechnique. McGraw-Hill Book Co., New York 1940.
    19. Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures. - Physiol. Plant. 15: 473-497, 1962. Go to original source...
    20. Nas, M.N.: Inclusion of polyamines in the medium improves shoot elongation in hazelnut (Corylus avellana L.) micropropagation. - Turk. J. Agr. Forest. 28: 189-194, 2004.
    21. Park, S.-Y., Ahn, J.-K., Lee, W.-Y., Murthy, H.N., Paek, K.-Y.: Mass production of Eleutherococcus koreanum plantlets via somatic embryogenesis from root cultures and accumulation of eleutherosides in regenerants. - Plant Sci. 168: 1221-1225, 2005. Go to original source...
    22. Pati, P.K., Sharma, M., Sood, A., Ahuja, P.S.: Direct shoot regeneration from leaf explants of Rosa damascena Mill. - In Vitro cell. dev. Biol. Plant 40: 192-195, 2004. Go to original source...
    23. Purohit, S.D., Singhvi, A., Nagori, R., Vyas, S.: Polyamines stimulates shoot bud proliferation and multiplication in Achras sapota grown in culture. - Indian J. Biotechnol. 6: 85-90, 2007.
    24. Ramesh, K., Padhya, M.A.: In vitro propagation of neem, Azadirachta indica (A. Juss.), from leaf discs. - Indian J. exp. Biol. 28: 932-935, 1990.
    25. Sacande, M., Golovina, E.A., Van Aelst, A.C., Hoekstra, F.A.: Viability loss of neem (Azadirachta indica) seeds associated with memberane phase behavior. - J. exp. Bot. 52: 919-931, 2001. Go to original source...
    26. Singh, A.K., Chand, P.K., Patnaik, S., Chand, S.: Adventitious shoot organogenesis and plant regeneration from cotyledons of Dalbergia sisoo Roxb., a timber yielding tree legume. - Plant Cell Tissue Organ Cult. 68: 203-209, 2002. Go to original source...
    27. Vila, S.K., Gonzalez, A.M., Rey, H.Y., Mroginski, L.A.: In vitro plant regeneration of Melia azedarach L.: shoot organogenesis from leaf explants. - Biol. Plant. 47: 13-19, 2003. Go to original source...
    28. Vila, S.K., Gonzalez, A.M., Rey, H.Y., Mroginski, L.A.: Plant regeneration, origin, and development of shoot buds from root segments of Melia azedarach L. (Meliaceae) seedlings. - In Vitro cell. dev. Biol. Plant 41: 746-751, 2005. Go to original source...
    29. Vinocur, B., Carmi, T., Altman, A., Ziv, M.: Enhanced bud regeneration in aspen (Populus tremula L.) roots cultured in liquid media. - Plant Cell Rep. 19: 1146-1154, 2000. Go to original source...
    30. Williams, J.G., Kubelik, A.R., Livak, K.J., Rafalski, J.A., Tingey, S.V.: DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. - Nucl. Acids Res. 18: 6531-6535, 1990. Go to original source...
    31. Yeoman, M.M., Street, H.E.: General cytology of cultured cells. - In: Street, H.E. (ed.): Plant Tissue and Cell Culture Vol. 2. Pp. 137-176. Blackwell Scientific Publications, Oxford 1977.

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