| Epicotyls from Citrus mitis seedling were used as explants to investigate the factors which had an influence on differentiation and regeneration of anti-Kan adventitious buds in transformation mediated by Agrobacterium. Among antibiotics, Timentin was the best for differentiation of adventitious buds and growth control of agrobacterium. Growing in darkness for 3~5 days after explants co-culture with agrobacterium was help to anti-Kan buds developing. It was better for anti-Kan adventitious buds developing when explants were cultured in differentiation medium for 2~3 days before infected by agrobacterium. Transgenic plants were obtained by PCR assay, and SOD enzyme activity and content of proline were raised in transgenic plant treated by -4益for 4~6 days. The result showed that transformation system was established for epicotyls in Citrus mitis. The transgenic plant could be obtain in medium with TDZ 0.5~1.0 mg/L, Timentin 300 mg/L, Km 100 mg/L. Frequency of transformation would be increased as explants precultured for 2~3 d and dark culture for 3~5 d after co-culture. |
