Factors Affecting Transformation Efficiency by Micro-Injecting Agrobacterium into Flower Bud of Chin

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Three main parameters were selected to study their importance in transformation by bud microinjection in non-head Chinese cabbage [Brassica campestris ssp. Chinensis (L.) Makino var. Communis Tsen et Lee]. The results showed that the developmental stage of floral bud, the concentrations of sucrose and surfactant Silwet L-77 were critical for the successful transformation by this new method. The suitable bud size is 2- 3 mm in length, the favorable concentration of sucrose and surfactant Silwet L-77 are 8 and 0.02% respectively. When the sucrose concentration was greater than 10% or that of Silwet L-77 was above 0.1%, the treated buds became yellow and finally blighted. 4/6 T1 seedlings resistant to kanamycin were positive by PCR analysis, and T2 progeny of all these positive T1 plants have one or more hybridizing bands by Southern blot. Under 5% sucrose, 0.02% Silwet L-77 and grade 2 bud (2- 3 mm in its length) parameters, the most favorable transformation efficiency is about 0.56%, and mean efficiency reaches 0.16% in all experiments indicating that bud microinjection is potential transformation way in non-head Chinese cabbage.
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