Plant Production Science
Online ISSN : 1349-1008
Print ISSN : 1343-943X
Improvement of Direct Somatic Embryogenesis in Rice by Selecting the Optimal Developmental Stage of Explant and Applying Desiccation Treatment
Totik Sri MARIANIHiroshi MIYAKEYoji TAKEOKA
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JOURNALS FREE ACCESS

2000 Volume 3 Issue 2 Pages 114-123

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Abstract

By selecting the optimal developmental stage of zygotic embryos used as explants and applying desiccation treatment, we improved direct somatic embryogenesis in rice scutellum from two cultivars, Nipponbare and Sasanishiki.Zygotic embryos isolated 14-17, 21, 28-30 and 35-40 d after anthesis(DAA)from Nipponbare and 14-17, 18-21, 28-30 and 40-42 DAA from Sasanishiki were cultured on the embryo induction medium(EIM).Then they were transferred to embryo maturation medium(EMM)and germinated on the embryo germination medium(EGM).Only immature zygotic embryo isolated 14-17 DAA from Nipponbare and Sasanishiki could develop somatic embryos that germinated.Explants from embryos at other developmental stages could develop somatic embryos only until the elongating or scutellar stage.They enlarged and formed callus without further development.The EIM and EMM consisted of N6 macronutrients, B5 micronutrients, and B5 vitamins, supplemented with 0.1 g L-1 casein hydrolysate, 1.5 g L-1 proline and 1 g L-1 MES buffer.EGM consisted of MS macro-and micronutrients and MS vitamins without organic supplement.In addition, 2 mg L-1 2, 4-D was added to EIM, 1 mg L-1 2, 4-D to EMM and 0.01 mg L-1 zeatin to EGM.Developmental processes of somatic embryos derived from the explants were observed by scanning electron microscopy.Desiccation treatment of maturing somatic embryo was proved to produce fully mature somatic embryos capable of germinating vigorously.

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