Lisianthus Leaf Regeneration System For Agrobacterium-mediated Npri Gene Conversion On A Lisianthus The Preliminary Study

Through the orthogonal experiments, a systematic research on the system of bud regenration from the leaf of Eustoma russellianum was carried out. The results indicated that the optimal culture midium for the bud differention was MS+ ZT 0.5mg/L+ NAA 0.01mg/L, pH value of medium was 6.3, the intensity of illumination was 60umol.m-2.s-1, The optimal culture medium for root induction was 1/4 MS+ IBA 1mg/L, containing sugar 50g/L, agar 4g/L, no active charcoal. The high efficiency shoot regeneration system from leaf has been established for the gene transformation of Eustoma russellianum.The suitable concentration of Hyg to screen for resistant shoots was 50 mg/L. The kind of antibiotics to inhibit the growth of Agrobacterium EHA105 was Cef, of which concentration was 250 mg/L. The result of study on the factors affecting the transformation frequency showed: the leaf disc was pre-cultured for 2 days, infected for 20 minutes, co-cultivated with Agrobacterium EHA105 for 2 days, which could improve regeneration efficiency. The highest regeneration efficiency was 13.3% in this experiment.Four Hyg-resistant plants, which were chosen randomly from the plants which grown in the field, were detected through GUS and PCR. The results confirmed that the NPR1 gene have been induced into the genome of Eustoma russellianum.