The genetic transformation was achieved by pollen_mediated approach on maize ( Zea mays L.) inbred lines Tai 9101 and Zong 31. Plasmid DNA of pGLⅡ_RC_1 was mixed with fresh pollen of maize inbreds in sucrose ...The genetic transformation was achieved by pollen_mediated approach on maize ( Zea mays L.) inbred lines Tai 9101 and Zong 31. Plasmid DNA of pGLⅡ_RC_1 was mixed with fresh pollen of maize inbreds in sucrose solution. The pollens were treated by ultrasonication and collected, pollinated on silks of maize ears. Transformants were confirmed by dot blot hybridization, PCR amplification and PCR_Southern blot hybridization. The pollen_mediated transformation approach could circumvent the tedious tissue culture procedures like in particle bombardment and Agrobacterium infection, etc. This approach is simple, easy to operate, and could be widely used in practice.展开更多
文摘The genetic transformation was achieved by pollen_mediated approach on maize ( Zea mays L.) inbred lines Tai 9101 and Zong 31. Plasmid DNA of pGLⅡ_RC_1 was mixed with fresh pollen of maize inbreds in sucrose solution. The pollens were treated by ultrasonication and collected, pollinated on silks of maize ears. Transformants were confirmed by dot blot hybridization, PCR amplification and PCR_Southern blot hybridization. The pollen_mediated transformation approach could circumvent the tedious tissue culture procedures like in particle bombardment and Agrobacterium infection, etc. This approach is simple, easy to operate, and could be widely used in practice.