Approaches to improve the transgenic efficiency and to rescue seedlings from hyperhydricity for rapeseed (Brassica napus)

摘要

Transgenic techniques play an important role in functional genomics research of Brassica napus, but an optimized, efficient transformation system has not been developed for this specie. The major difficulties come from the long regeneration period, low regeneration rate, and high hyperhydricity rate. In this study, we established a method with modifications on the callus-induction step, which was performed at the absence or low level of selective antibiotic in previous approaches. By adding selective antibiotic in callus-induction medium, the new method greatly enhanced the elimination of untransformed explants and shortened the period of seedling regeneration. We also established a convenient solution to rescue transgenic seedlings from hyperhydricity, a serious problem during the process of rapeseed transformation. The application of this method could produce transgenic seedlings ready for transplantation in less than eight weeks. Taken together, we have established an efficient, simple transformation system to produce transgenic rapeseed seedlings rapidly and efficiently.