EFFICIENT CALLUS INDUCTION AND PLANT REGENERATION FROM IMMATURE EMBRYO CULTURE OF TRITICUM AESTIVUM

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An efficient plant regeneration system was developed from the immature embryos of Triticum aestivum L. - Thinopyrum intermedium alien disomic addition lines, which resistant to powdery mildew. The protocol was based on a series of experiments involving the callus induction and differentiation. The experiment studied the effects of embryo size on callus induction and differentiation of the immature embryos. We found that the embryo size is critical for the establishment of embryogenic callus. Immature embryos (0.8~1.5 mm) showed high ability to produce embryogenic callus capable of regenerating green plants. The medium Murashige and Skoogs (MS) added with 2mg/L 2, 4-dichlorophenoxyacetic acid (2, 4-D) gave the best embryogenic callus induction, maintenance and regeneration. The embryogenic callus maintained high regeneration during six subcultures in the callus induction medium. Suitable time of partial desiccation could effectively improve the regeneration capacity of the callus cultured for 3~4 month.Bud green spot and root green spot were observed during the differentiation of callus and the difference between them was described. Regenerated shoots were rooted on half-strength MS medium containing 0.2 mg/L Naphthalene acetic acid (NAA). Plants were successfully transferred to soil and grew well. This efficient plant regeneration system provides a foundation for the study of somaclonal variation of Triticum aestivum L. - Thinopyrum intermedium alien disomic addition lines.
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