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Role of phytohormones and nitrogen in somatic embryogenesis induction in cell culture derived from leaflets of Azadirachta indica

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Biologia Plantarum


A protocol for somatic embryogenesis in Azadirachta indica A Juss. has been standardized using in vivo leaflets. Experiments were carried out to examine the effect of various auxins, cytokinins, sucrose, inorganic and organic salts on subsequent somatic embryo induction and maturation. Embryogenic calli were induced on Murashige and Skoog (MS) medium supplemented with 1.5 mg dm−3 kinetin and 1.5 mg dm−3 indole-3-acetic acid and subsequently all the stages of somatic embryo development (globular, cordate, torpedo and cotyledonary) were observed. Maturation of these embryos was accomplished with the same growth regulators after three subcultures. The histological study of somatic embryos showed resembles to zygotic embryos. The matured somatic embryos were transferred onto half strength MS-medium devoid of growth regulators for their germination (82 %). Plantlets were acclimatized in the field with a survival rate of 80–83.5 %.

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2,4-dichlorophenoxyacetic acid


N 6-benzylaminopurine


indole-3-acetic acid


indole-3-butyric acid




Murashige and Skoog medium


naphthaleneacetic acid


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Correspondence to G. S. Shekhawat.

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Acknowledgement: G.S. Shekhawat greatly acknowledges the grant received from CSIR, New Delhi, as Senior Research fellow.

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Shekhawat, G.S., Mathur, S. & Batra, A. Role of phytohormones and nitrogen in somatic embryogenesis induction in cell culture derived from leaflets of Azadirachta indica . Biol Plant 53, 707–710 (2009).

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