Establishment of efficient transformation system of Brassica parachinensis mediated by Agrobacterium tumefacien

Based on the established regeneration system,an efficient transformation and screening system of Brassica parachinensis mediated by Agrobacterium was developed.Four-day-old cotyledon served as excellent explants for transformation,and the factors affecting the transformation frequency,such as the kinds and concentrations of antibiotics to inhibit the growth of Agrobacterium,Agrobacterium strains,preculture time of cotyledon explants,concentration of Agrobacterium,infection time and co-infection time etc.were also discussed by GUS coloration analysis.2 days preculturation,15 min Agrobacterium(OD_(600)=0.8) infection with 100 μmol/L acetosyringone(AS),and 2 days' co-infection would lead to an increase in transformation efficiency.The differential medium with kanamycin 15 mg/L and Ticarcillin 100 mg/L was found to be most optimal for transgenic plantlets selection.Plantlets resistant to kanamycin were obtained and 8 transgenetic plants were obtained.Stable integration of GUS gene in these plants were confirmed by PCR and Southern blotting analysis.