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Somatic Embryogenesis from Immature Zygotic Embryos of Sterculia alata Roxb. and Monitoring the Ontogeny and the Genetic Fidelity of Somatic Embryos

Madhu Tripathi, Nishi Kumari, Vijay S. Jaiswal, Uma Jaiswal, Major Singh

Abstract


A protocol for the induction of somatic embryogenesis from immature zygotic embryos of Sterculia alata was established. The highest induction frequency for somatic embryogenesis was achieved using 3 mm size of zygotic embryos as explants on Murashige and Skoog (MS) medium supplemented with 2,4-D (2.0 mgl–1) + BAP (0.1 mgl–1) and 3% sucrose. The embryogenic calli differentiated when transferred to MS basal medium (maintenance medium) after three weeks treatment in induction medium. Frequency of embryogenesis was enhanced by the addition of organic nitrogen (glutmine, proline) in the maintenance medium but optimum frequency was observed by the supplementation of 400 mgl–1 glutamine. The best medium for embryo maturation was MS medium containing 3% sucrose and 0.01 mgl–1 ABA. Approximately 64% of selected mature SEs were able to germinate in the modified WPM having 3% sucrose but the conversion frequency was very low and it was only 7.07%. Histology of somatic embryos showed that fusion of somatic embryo is the main reason which hinders the conversion process. The random amplified polymorphic DNA was used to analyze genetic fidelity of different stages of somatic embryos and also to identify the problem in conversion if any at genetic level. Somatic embryos of different stages showed no aberration in RAPD banding patterns with respect to donor explants.

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