03Apr 2019

OPTIMIZATION OF TISSUE CULTURE PROTOCOL FOR REGENERATION AND DIRECT MULTIPLICATION OF MORINGA OLEIFERA BY SEED GERMINATION.

  • Shaheed Zulfikar Ali Bhutto Institute of Science and Technology (SZABIST) Department of Biosciences, Karachi 75600, Pakistan.
  • Abstract
  • Keywords
  • References
  • Cite This Article as
  • Corresponding Author

Moringaoleifera(G.H.No.:95436,Dr.MuneebaKhan,CentreforPlantConservation,University of Karachi) known as the miracle tree, is a plant of great importance. The plant is well known for its diverse medicinal and nutritional properties. The objective of the research is focused on tissue culture of Moringaoleifera through callus induction, direct multiplication as well as from seed culture.Thestudyisalsofocusedonoptimizationofsterilizationprotocolandmediaforthetissue culture processes. As expected, Moringaoleifera gave promising results using MS Media supplementedwithdifferentconcentrationsofPlantGrowthRegulatorssuchasIAA,NAA,BAP, Kinetin and 2,4-D. Seed initiation gave optimum results on MS Basal media. Direct multiplication of the plant was observed with optimum results on MS media supplemented with 0.1 mgL-1BAP. For callus induction, effects of Plant Growth Regulator and photoperiod were observed. In Dark conditions, maximum growth was seen on MS media supplemented with 2, 4-D (2 mgL-1), and that in light conditions, MS media with 2,4-D (2 mgL-1and 3 mgL-1) gave almost similar results. Initially, seed coats were removed inside the Laminar Flow Hood but this was affecting the sterilization negatively. Seed sterilization was then modified by removing seed coats and sterilizing them outside the Laminar Flow Hood. The seeds were then transferred to the LFH for furthersterilization,andthenculturedonthe growthmedia.Forthesterilizationoftheexplantfor directmultiplicationandcallusinduction,firstlyEthanolwasusedwhichcausedbleachingeffect. Later, Ethanol was removed from the protocol, reducing the bleaching effect to zero, and sterilization was maximized.


  1. Ahloowalia,B.S.,Prakash,J.,Savangikar,V.A. and Savangikar,C.(2004):Planttissueculture. Low costoptionsfortissue culture technology in developing countries. Int At Energy Agency Vienna.,3-11.
  2. Al Khateeb, W., Bahar, E., Lahham, J., Schroeder, D. and Hussein, E. (2013): Regeneration and assessment of genetic fidelity of the endangered tree Moringaperegrina(Forsk.) Fiori using Inter Simple Sequence Repeat (ISSR). PhysiolMolBiol Plants., 19: 157-164.
  3. Anwar, F., Latif, S., Ashraf, M. and Gilani, A.H. (2007): Moringaoleifera: a food plant with multiple medicinal uses. Phytother Res., 21: 17-25.
  4. Balunas, M.J. and Kinghorn, A.D. (2005): Drug discovery from medicinal plants. Life Sci., 78: 431-441.
  5. Banerji, R., Bajpai, A. and Verma, S.C. (2009): Oil and fatty acid diversity in genetically variable clones of Moringaoleiferafrom India. J Oleo Sci., 58: 9-16.
  6. Bennett,R.N.,Mellon,F.A.,Foidl,N.,Pratt,J.H.,Dupont,M.S.,Perkins, andKroon,P.A.(2003):Profilingglucosinolatesand phenolicsinvegetativeandreproductivetissuesofthemulti-purposetreesMoringaoleiferaL.(horseradishtree) and MoringastenopetalaL. J Agric Food Chem., 51:3546-3553.
  7. Bichi, M.H. (2013): A review of the applications of Moringaoleiferaseeds extract in water treatment. CivEnviron Res., 3: 1-10.
  8. Debergh, P.C. and Read, P.E. (1991): Micropropagation. In Micropropag Springer Neth,. : 1-13.
  9. Faizi, S., Siddiqui, B.S., Saleem, R., Siddiqui, S. and Aftab, K. (1995): Fully acetylated carbamate and hypotensive thiocarbamate glycosides from Moringaoleifera. Phytochem,. 38: 957-963.
  10. Ferreira, P.M.P., Farias, D.F., Oliveira, J.T.D.A. and Carvalho, A.D.F.U. (2008): Moringaoleifera: bioactive compounds and nutritional potential. Revista de Nutri??o,. 21: 431-437.
  11. F?rster, N., Mewis, I. and Ulrichs, C. (2013): MoringaOleifera?Establishment and Multiplication of Different Ecotypes in vitro. GesundePflanzen., 65: 21-31.
  12. Fuglie, L.J. (1999): The Miracle Tree: Moringaoleifera: Natural Nutrition for the Tropics. Church World Serv, Dakar., 68.
  13. Gassenschmidt, U., Jany, K.D., Bernhard, T. and Niebergall, H. (1995): Isolation and characterization of a flocculating protein from Moringaoleifera Biochimica et BiophysicaActa (BBA)-Gen Subj., 1243: 477-481.
  14. Islam, S., Jahan, M.A.A. and Khatun, R. (2005): In vitro regeneration and multiplication of year-round fruit bearing
  15. Moringaoleifera J Biol Sci., 5: 145-148.
  16. Jaskani, M.J., Abbas, H., Khan, M.M., Qasim, M. and Khan, I.A. (2008): Effect of growth hormones on micropropagation of Vitisvinifera cv. Perlette. Pak J Bot., 40: 105.
  17. Kadhimi, A.A., Alhasnawi, A.N., Mohamad, A., Yusoff, W.M.W. and Zain, C. (2014): Tissue culture and some of the factors affecting them and the micropropagation of strawberry. Life Sci J., 11: 484-493.
  18. Kinghorn, A.D. (2001): Pharmacognosy in the 21st century. J Pharm Pharmacol., 53: 135-148. Marfori, E.C. (2010): Clonal micropropagation of Moringaoleifera Philipp Agric Sci., 93:454-457.
  19. Mathur, M., Yadav, S., Katariya, P.K. and Kamal, R. (2014): In vitro propagation and biosynthesis of steroidalsapogenins from various morphogenetic stages of Moringaoleifera, and their antioxidant potential. ActaPhysiolPlant., 36: 1749-1762.
  20. Murakami, T., Ise, K., Hayakawa, M., Kamei, S. and Takagi, S.I. (1989): Stabilities of metal complexes of mugineic acids and their specific affinities for iron (III). ChemLett., 18: 2137-2140.
  21. Murashige, T. and Skoog, F. (1962): A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant., 15: 473-497.
  22. Palada, M.C. (1996): Moringa (Moringaoleifera): a versatile tree crop with horticultural potential in the subtropical United States. HortSci., 31: 794-797.
  23. Pari, L. and Kumar, N.A. (2002): Hepatoprotective activity of Moringaoleiferaon antitubercular drug-induced liver damage in rats. J Med Food., 5: 171-177.
  24. Preece, J.E. and Read, P.E. (1993): The biology of horticulture: an introductory textbook. John Wiley &Son.
  25. Riyathong, T., Dheeranupattana, S., Palee, J. and Shank, L. (2010): Shoot multiplication and plant regeneration from in vitro cultures of Drumstick tree (Moringaoleifera). The IntSympBiocontrol Biotech., 154-159.
  26. Saini,R.K.,Shetty,N.P.,Giridhar,P. andRavishankar,G.A.(2012):RapidinvitroregenerationmethodforMoringaoleiferaand performance evaluation of field grown nutritionally enriched tissue cultured plants. 3 Biotech., 2: 187-192.
  27. Shahzad, U., Khan, M.A., Jaskani, M.J., Khan, I.A. and Korban, S.S. (2013): Genetic diversity and population structure of Moringaoleifera. Conserv Genet., 14: 1161-1172.
  28. Shahzad, U., Jaskani, M.J., Ahmad, S. and Awan, F.S. (2014): Optimization of the micro-cloning system of threatened Moringaoleifera Pak J Agric Sci., 51: 449-457.
  29. Stephenson, K.K. and Fahey, J.W. (2004): Development of tissue culture methods for the rescue and propagation of endangered Moringa spp. Econ Bot., 58: S116-S124.
  30. Verdcourt, B. (1985): A synopsis of the Moringaceae. Kew Bull., 40: 1-23.
  31. Wagner, H. and Bladt, S. (1996): Plant drug analysis: a thin layer chromatography atlas. Springer Sci Bus Media.

[Asfandyar Chaudhary, Kashif Ali, Neha Farid And Nosheen Maqsood. (2019); OPTIMIZATION OF TISSUE CULTURE PROTOCOL FOR REGENERATION AND DIRECT MULTIPLICATION OF MORINGA OLEIFERA BY SEED GERMINATION. Int. J. of Adv. Res. 7 (Apr). 547-555] (ISSN 2320-5407). www.journalijar.com


Kashif Ali
Department of Biosciences, Shaheed Zulfikar Ali Bhutto Institute of Science and Technology, Karachi, Pakistan.

DOI:


Article DOI: 10.21474/IJAR01/8851      
DOI URL: https://dx.doi.org/10.21474/IJAR01/8851