Study On Pollen-mediated Transformation Of Rapeseed(Brassica Napus L.) For Breeding

Plasmid DNA of pBI121 were mixed with fresh pollen of rapeseed lines, Jinyou 7, in 7.5% saccharose solution, respectively. The pollen suspension were treated with ultrasonication and collected, pollinated on stigmas of rapeseed. Transformants were confirmed by field comparetion, histochemical examination of GUS expression and PCR amplification. 6 transgenetic T1 plants were obtained, and the transformation rate was 2.8%. In the study, some regeneration conditions, such as the thickness of saccharose solution, the intensity of ultrasonic and the effective of auxiliary for transformation system were discussed. 7.5% of saccharose solution or 5% of manna sugar, 300W of ultrasonic were perfect. The highly efficient transformation system on rapeseed was established. Compare with other transformations, the pollen-mediated transformation approach could circumvent the tedious tissues culture procedures like in Agrobacterium infection and particle bombardment, etc. This approach is simple, easy to operate, and could be widely used in breeding practice.