In vitro direct organogenesis and regeneration of Medicago sativa
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A rapid and efficient plant regeneration protocol for a wide range of alfalfa genotypes was developed via direct organogenesis. Through a successive excision of the newly developed apical and axillary shoots, a lot of adventitious buds were directly induced from the cotyledonary nodes when hypocotyl of explants were vertically inserted into modified Murashige and Skoog (MS) medium supplemented with 0.025 mg dm−3 thidiazuron (TDZ) and 3 mg dm−3 AgNO3. When the lower part of shoots excised from explants were immersed into the liquid medium with 1.0 mg dm−3 α-naphthaleneacetic acid (NAA) for 2 min, and then transferred to hormone free half-strength MS medium, over 83.3 % of the shoots developed roots, and all plantlets could acclimatize and establish in soil. The protocol has been successfully applied to eight genotypes, with regeneration frequencies ranging from 63.8 to 82.5 %.
Additional key wordsalfalfa cotyledonary node mature embryo silver nitrate thidiazuron vitrification
Murashige and Skoog
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