大红甜橙遗传转化高效再生体系的建立

Using epicotyl and hypocotyl pieces and cotylcdons of Citrus sinensis Osbeck cv．Dahong seedlings grown in vitro as explants，an efficient regeneration system was establlished. The optimal conditions were obtained as follows. 1) The epicotyl pieces were used as explants．2)The optimal advendtitious shoot regeneration medium was MS salts+100mg/L inositol +0.2 mg/L V-B1+1 mg/L V-B6+1 mg/L niacin +3%sucrose +0.8% agar +3 mg/L 6-BA, pH5.8. The best response was obtained when tIhe explants were incubated at 26℃ for 7days in darkness, followed by 33 days on a16/8 h ligh/dafk cycle, adventitious shoot regenerated from 90% of the epicotyl explants. The best adventitious root regeneration medium was 1/2MS+3％sucrose+0.7％ agar 2.0 mg/L IBA, pH5.8. Adventitious root regenerated from 94.7%of the adventitious shoot after growing in vitro for 40 d. 3)Tlhe suitable selection scheme was culturing the epicotyl pieces on the adventitious shoot regeneration medium containing 50 mg/L Km．