Agrobacterium-mediated Transformation of Ground-cover Chrysanthemum(Dendranthema grandiflorum cv. White Snow) with Transcription Factor DREB1A Gene

Using in vitro leaves as explants, the adventitious shoots of ground-cover chrysanthemum(Dendranthema grandiflorum cv. White Snow) were derived from the basal MS media supplemented with different plant growth regulators through organogenesis. The results indicated that the regeneration rate of 93.8% was obtained by the combination of 2.0 mg/L 6-BA and 2 mg/L NAA. The binary vector pBI-DREB1A containing transcription factor DREB1A gene from Arabidopsis thaliana was introduced into Agrobacterium tumefaciens LBA4404. The leaf disks of ground-cover chrysanthemum, White Snow, were infected through Agrobacterium-mediated transformations. The DREB1A gene was integrated into the genome of transgenic plants confirmed by PCR and PCR-Southern analysis. The highest transformation efficiency was obtained through the following transformation procedure: after preculture for 2 d, the explants were infected for 10 min with diluted OD600=0.5~0.7 Agrobacterium tumefaciens culture liquid with a ratio of 1/30, and then co-cultivated for 2 d.