Regeneration of Complete Plantlets from Callus Culture of Azadirachta indica A. Juss using Immature Flower Buds
DOI:
10.34062/afs.v4i1.3978Keywords:
Azadirachta indica, flower buds, callus cultureAbstract
Plant regeneration via callus culture was achieved using immature flower buds of mature tree of Azadirachta indica A. Juss on semi-solid MS medium enriched with 9.05µM 2,4-D + additives. The regenerated callus were amplified on lower concentration of 4.52 µM 2,4-D + additives for micro-shoot regeneration study, the best micro-shoot regenerated from callus was achieved on MS medium supplemented with 8.88 µM BAP with 3.78±0.50 numbers of shoot and 3.66±0.57 shoot length. The micro-shoots were amplified on 8.88 µM BAP+ 0.27 µM NAA with 5.22±0.53 numbers of shoot and 4.62±0.59 shoot length respectively. Micro shoot were successfully rooted on MS medium supplemented with IBA 4.92 µM with rooting percent (100%), 4.35±0.22 root number and 3.95±0.23 root length. The callus derived plantlets were hardened in vitro and ex vitro and acclimatized in poly house followed by agro-shade house. The efficiency of the protocol will be helpful for propagation of Azadirachta indica plants on a mass scale.
References
Allan EJ, Easwara JP, Wood A (1994) The production of azadirachtin by in vitro tissue culture of neem, Azadirachta indica. Pesti. Sci. 42: 147-152.
Carvalho DC, Silva ALL, Schuck MR, Purcino M, Tanno GN, Biasi LA (2013) Fox grape cv. Bordô (Vitis labrusca L.) and grapevine cv. Chardonnay (Vitis vinifera L.) cultivated in vitro under different carbohydrates, amino acids and 6-benzylaminopurine levels. Braz Arch Biol Technol, 56: 191-201.
Eeswara JP, Stuchbury T, Allan EJ, Mordue AJ (1998) A standard procedure for the micropropagation of the neem tree (Azadirachta indica A. Juss). Plant Cell Rep. 17: 215–219
Ermel K, Pahlich E, Schmutterer H (1986) Azadirachtin content of neem kernels from different geographical locations, its dependence on temparature, relative humidity and light. In: Schmutterer H, Ascher KRS (eds) Natural pesticides from the neem tree (Azadirachta indica A. Juss) and other tropical plants. In: Proceedings of the Third International Neem Conference. GTZ, Eschborn, pp 171–184
Gautam VK, Nanda K, Gupta SC (1993) Development of shoots and roots in anther-derived callus of Azadirachta indica A. Juss. A medicinal tree. Plant Cell Tissue Organ Culture. 34: 13–18.
Gill AS, Roy RD (1996) Agroforestry Uses, In: Randhawa N.S., Parmar B.S. (eds) Neem, New Age International (P) Ltd, New Delhi, 49-62.
Jacobson M (1988) Focus on phytochemical pesticides, vol 1. The neem tree. CRC, Boca Raton, Fla.
Johnson S, Morgan, ED, Wilson ID, Spraul M, Hofmann M (1994) Photo-isomerization of azadirachtin by high-performance liquid chromatography coupled to high-field proton NMR-spectroscopy. Journal Chem Soc Perkin Trans 1:1499–1502.
Joshi MS, Thengane SR (1996) In vitro propagation of Azadirachta indica A. Juss (neem) by shoot proliferation. Ind. J. Exp. Biol. 5: 480–482.
Kaushik N (2002) Determination of azadirachtin and fatty acid methyl esters of Azadirachta indica seeds by HPLC and GLC, Anal. Bioanal. Chem., 374, 199-204.
Kearney ML, Allan EJ, Hooker J, Mordue (Luntz) AJ, (1994) Antifeedant effects of in vitro culture extracts of the neem tree, Azadirachtaindica against the desert locust [Schistocerca gregaria (Forskål)]. Plant Cell Tissue Organ Cult 37:67–71.
Koul O, Isman MB, Ketkar CM (1990) Properties and Uses of neem, Azadirachta indica, Can. J. Bot., 68, 1-11.
Litz RE, Conover RA (1981) Effect of sex type, season and other factors on in vitro establishment and culture of Carica papaya L. explants, J. Amer. Soc. Hort. Sci.,106, 792-794.
Larson LL (1993) Materials under trial: information on composition, formulation and suppliers. Insecticide Acaricide Tests 18:375–390.
Ley SV, Denholm AA, Wood A (1993) The chemistry of azadirachtin. Nat Prod Rep :109–157.
Mohan Ram HY, Nair MNB (1996) Botany. In: Randhawa N.S. and Parmar B.S. (eds) Neem, New Age International (P) Ltd, New Delhi, 6-26.
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 15: 473–497.
Muralidharan EM, Mascarenhas AF (1989) In vitro morphogenesis in Azadirachta indica A. Juss and Eucalyptus citriodora Hook. In: Kukreja AK, Mathur AK, Ahuja PS, Thakur AS (eds). Tissue Culture and Biotechnology of Medicinal and Aromatic Plants. New Delhi:Paramount Publishers, pp. 49-55.
Murthy BN, Saxena P (1998) Somatic embryogenesis and plant regeneration of neem (Azadirachta indica A. Juss.). Plant Cell Rep. 17:469–475.
Narayan P, Jaiswal VS (1985) Plantlet regeneration from leaflet callus of Azadirachta indica Juss. J. Tree Sci., 4: 65-68.
National Research Council (1992) Neem: a tree for solving global problems. National Academy Press, Washington, DC.
Quraishi A, Koche V, Sharma P, Mishra SK (2004) In vitro clonal propagation of neem (Azadirachta indica). Plant Cell Tiss. Organ Cult. 78: 281– 284.
Ramesh K, Padhya MA (1990) In vitro propagation of Neem, Azadirachta indica (A. Juss), from leaf disks. Indian Journal Exp. Biol. 28: 932–935.
Salvi ND, Singh H, Tivarekar S, Eapen S (2001) Plant regeneration from different explants of neem. Plant Cell Tiss. Organ Cult. 65: 159– 162.
Schmutterer H (1990) Properties and potential of natural pesticides from the neem tree, Azadirachta indica. Annu Rev Entomol 35:271–297.
Singh RP (1986) Comparison of antifeedant efficacy and extract yields form different parts and ecotypes of neem (Azadirachtaindica A. Juss) trees. In: Schmutterer H, Ascher KRS (eds) Natural pesticides from the neem tree (Azadirachta indica A. Juss) and other tropical plants. Proceedings of the Third International Neem Conference. GTZ, Eschborn, pp 185–194.
Sanyal M, Banerjee M, Dutta PC (1981) In vitro hormone induced chemical and histological differentiation in stem callus of Neem (Azadirachta indica A.juss). Indian Journal of Experimental Biology. 19,1067-1068.
Snedecor GW, Cochran WG (1967) Statistical Methods. The Iowa State University Press, Iowa USA. pp. 327–9.
Shrikhande M, Thengane SR, Mascarenhas AF (1993) Somatic embryogenesis and plant regeneration in Azadirachta indica A. Juss. In Vitro Cell. Dev. Biol. Plant 29:38–42.
Silva ALL, Costa JL, Gollo AL, Santos JD, Forneck HR, Biasi LA, Soccol VT, Carvalho JC, Soccol CR (2014) Development of a vinasse culture medium for plant tissue culture. Pak. J. Bot., 46(6): 2195-2202.
Su WW, Hwang WI, Kim SY, Sagawa Y (1997) Induction of somatic embryogenesis in Azadirachta indica. Plant Cell Tiss. Organ Cult. 50:91–95.
Tewari DN (1992) Monograph on Neem (Azadirachta indica A. Juss). International Book Distributors, Dehradun, 279.
Thengane S (1995) Somatic Embryogenesis in Neem (Azadirachta indica). In: Jain S.M., Gupta P., Newton R. (eds) SomaticEmbryogenesis in Woody Plants. Vol. II. Important Selected Plants.Kluwer academic Publishers, Dordrecht, 357-374
Yasseen YM (1994) Shoot proliferation and plant formation from neem with thidiazuron. Hortscience 29:515;
Zypman S, Ziv M, Applebaum S (1997) Tissue culture methods and cloning of the neem tree (Azadirachta indica) for bioinsecticide production. Acta Hortic. 447: 235–236.
Downloads
Published
Issue
Section
License
All copyright must be assigned to the Federal University of Mato Grosso.
