An efficient procedure was described for the transformation of the monocotyledonous oriental hybrid lily, Lilium cv. Siberia. by Agrobacterium-mediated genetic transformation via leaves regeneration, The leaves of lea...An efficient procedure was described for the transformation of the monocotyledonous oriental hybrid lily, Lilium cv. Siberia. by Agrobacterium-mediated genetic transformation via leaves regeneration, The leaves of leaflets which derived from bulbs were sliced into 1.0 cm long and were co-cultivated with A. tumefaciens strain LBA4404/pB2AE12, which harbored a vector carrying the neomycin phosphotransferase, DREB2A genes in the T-DNA region. The suitable genetic transformation condition was determined as follows： the bacterial concentration reached 0.5-0.6 （OD600）, 15 min infection time, 20 mg.L^-1 acetosyingone, and 10.6 mmol.L^-1 NH4NO3 medium was used for co-cultivation 3 days, delayed 7 days for selecting by 30 mg.L^-1 kanamycin containing regeneration medium. Efficient shoot regeneration was observed on MS medium supplemented with 1.0 mg.L^-1 naphthaleneacetic acid, 0.5 mg.L^-1 benzyladenine and 0.1 mg.L^-1 Kinetin after about 6 weeks culture. The presence ofDREB2A gene in the genomic DNA of regenerated plants was detected by means of PCR analysis.展开更多
基金Supported by Postdoctoral Fund of Settling Down in Heilongjiang Province (LBH-Z08259)Program for Dr. Research Fund of Northeast Agricultural University
文摘An efficient procedure was described for the transformation of the monocotyledonous oriental hybrid lily, Lilium cv. Siberia. by Agrobacterium-mediated genetic transformation via leaves regeneration, The leaves of leaflets which derived from bulbs were sliced into 1.0 cm long and were co-cultivated with A. tumefaciens strain LBA4404/pB2AE12, which harbored a vector carrying the neomycin phosphotransferase, DREB2A genes in the T-DNA region. The suitable genetic transformation condition was determined as follows： the bacterial concentration reached 0.5-0.6 （OD600）, 15 min infection time, 20 mg.L^-1 acetosyingone, and 10.6 mmol.L^-1 NH4NO3 medium was used for co-cultivation 3 days, delayed 7 days for selecting by 30 mg.L^-1 kanamycin containing regeneration medium. Efficient shoot regeneration was observed on MS medium supplemented with 1.0 mg.L^-1 naphthaleneacetic acid, 0.5 mg.L^-1 benzyladenine and 0.1 mg.L^-1 Kinetin after about 6 weeks culture. The presence ofDREB2A gene in the genomic DNA of regenerated plants was detected by means of PCR analysis.
