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An Efficient Protocol for Cyclic Somatic Embryogenesis in Neem (Azadirachta indica A Juss.)
Abstract:Neem is a highly heterozygous and commercially important perennial plant. Conventionally, it is propagated by seeds which loose viability within two weeks. Strictly cross pollinating nature of the plant causes serious barrier to the genetic improvement by conventional methods. Alternative methods of tree improvement such as somatic hybridization, mutagenesis and genetic transformation require an efficient in vitro plant regeneration system. In this regard, somatic embryogenesis particularly secondary somatic embryogenesis may offer an effective system for large scale plant propagation without affecting the clonal fidelity of the regenerants. It can be used for synthetic seed production, which further bolsters conservation of this tree species which is otherwise very difficult The present report describes the culture conditions necessary to induce and maintain repetitive somatic embryogenesis, for the first time, in neem. Out of various treatments tested, the somatic embryos were induced directly from immature zygotic embryos of neem on MS + TDZ (0.1 μM) + ABA (4 μM), in more than 76 % cultures. Direct secondary somatic embryogenesis occurred from primary somatic embryos on MS + IAA (5 μM) + GA3 (5 μM) in 12.5 % cultures. Embryogenic competence of the explant as well as of the primary embryos was maintained for a long period by repeated subcultures at frequent intervals. A maximum of 10 % of these somatic embryos were converted into plantlets.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1079470Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1180
 C.J.J.M. Raemakers, E. Jacobsen, and R.G.F. Visser, Secondary somatic embryogenesis and application in plant breeding. Euphytica, 1995, 81: 93-107.
 S.A. Merkle, W.A. Parrott, and E.G. Williams, Applications of somatic embryogenesis and embryo cloning. In: Bhojwani SS (ed) Plant tissue culture: applications and limitations. Elsevier, Amsterdam, 1990, pp 67- 101.
 C. Akula, A. Akula, and R. Drew, Somatic embryogenesis in clonal neem, Azadirachta indica A. Juss. and analysis for in vitro azadirachtin production. In Vitro Cell Dev-Pl., 2003, 39:304-310.
 R. Chaturvedi., M.K. Razdan, and S.S. Bhojwani, In vitro morphogenesis in zygotic embryo cultures of neem (Azadirachta indica A. Juss.) Plant Cell Rep., 2004, 22:801-809.
 A. Gairi, and A. Rashid, Direct differentiation of somatic embryos on different regions of intact seedlings of Azadirachta in response to thidiazuron. J. Plant Physiol., 2004a, 161:1073-1077.
 Gyana Ranjan Rout, In vitro somatic embryogenesis in callus cultures of Azadirachta indica A. Juss.ÔÇöa multipurpose tree. J. For. Res., 2005, 10:263-267.
 T. Murashige, and F. Skoog, A revised medium for rapid growth and bioassays with tobacco tissue cultures. - Physiol. Plant, 1962, 15: 473- 497.
 E.G. Williams, and G. Maheswaran, Somatic embryogenesis: factors influencing coordinated behaviour of cells as an embryogenic group. Ann. Bot., 1986, 57:443-462.
 O. Karami, A. Deljou, and A.M. Pour, Repetitive somatic embryogenesis in carnation on picloram supplemented media. Plant Growth Regul., 2007, 51:33-39.
 R.R. Nair, and S.D. Gupta, High-frequency plant regeneration through cyclic secondary somatic embryogenesis in black pepper (Piper nigrum L.) Plant Cell Rep., 2006, 24: 699-707.
 S. Agarwal, K. Kanwar, and D.R. Sharma, Factors affecting secondary somatic embryogenesis and embryo maturation in Morus alba L. Sci. Hort., 2004, 102: 359-368.
 P. Giridhar, E.P. Indu, G.A. Ravishankar and A. Chandrasekar, Influence of triacontanol on somatic embryogenesis in Coffea arabica L. and Coffea canephora P. ex. Fr. In Vitro Cell Dev Biol Plant, 2004, 40:200- 203.
 J.-T, Chen, and W.-C. Chang, Induction of repetitive embryogenesis from seed-derived protocorms of Phalaenopsis amabilis var. Formosa Shimadzu. In Vitro Cell Dev Biol Plant, 2004, 40:290-293.
 E.L, Little, Z.V. Magbanua, and W.A. Parrott, A protocol for repetitive somatic embryogenesis from mature peanut epicotyls. Plant Cell Rep., 2000, 19:351-357.
 L.O. das Neves, S.R.L. Duque, J.S. de Almeida and P.S. Fevereiro, Repetitive somatic embryogenesis in Medicago truncatula sp. Narbonensis and M. truncatula Gaertn cv. Jemalong. Plant Cell Rep., 1999, 18:398-405.
 R. Parra, and J.B. Amo-Marco, Secondary somatic embryogenesis and plant regeneration in myrtle (Myrtus communis L.). Plant Cell Rep., 1998, 18:325-330.
 P. Das, S. Samantaray, A.V. Roberts, and G.R, Rout, In vitro somatic embryogenesis of Dalbergia sisoo Roxb.ÔÇöa multipurpose timberyielding tree. Plant Cell Rep., 1997, 16:578-582.
 G. Daigny, H. Paul, R.S. Sangwan, and B.S. Sangwan-Narreel, Factors influencing secondary somatic embryogenesis in Malus x domestica Borkh. (cv. ÔÇÿGloster 69-). Plant Cell Rep., 1996, 16:153-157.
 F. Carimi, F. De Pasquale, and F.G. Crescimanno, Somatic embryogenesis and plant regeneration from pistil thin cell layers of Citrus. Plant Cell Rep., 1999, 18:935-940.
 J.-T, Chen, and W.-C. Chang, Effect of auxins and cytokinins on direct somatic embryogenesis on leaf explants of Oncidium ÔÇÿ Gower Ramsey-. Plant Growth Regul., 2001, 34: 229-232.
 D. calic, S. Zdravkovic-Korac, and Lj. Radojevic, Secondary embryogenesis in androgenic embryo cultures of Aesculus hippocastanum L. Biol. Plant, 2005, 49:435-438.
 P. Pui Puigderrajols, C. Celestino, M. Sulis, M. Toribio, and M. Molinas,. Histology of organogenic and embryogenic responses in cotyledons of somatic embryos of Quercus suber L. Int. J. Plant Sci., 2000, 161: 353-362.
 W.W. Su, W.I. Hwang, S.Y. Kim, and Y. Sagawa, Induction of somatic embryogenesis in Azadirachta indica. Plant Cell Tissue Organ Cult., 1997, 50:91-95.