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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· ets 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. Effi cient 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 of DREB2A gene in the genomic DNA of regenerated plants was detected by means of PCR analysis.
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. Ets 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 for 3 days, delayed 7 days for selecting by 30 mg · L-1 kanamycin containing regeneration medium. 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 of DREB2A gene in the genomic DNA of regenerated plants was detected by means of PCR analysis.