Haploid transformation in Brassica napus using an octopine-producing strain of Agrobacterium tumefaciens.
作者： E. B. Swanson , L. R. Erickson
DOI: 10.1007/BF00266666
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摘要: Microspore-derived embryos of Brassica napus were transformed using the disarmed octopine-producing LBA4404 strain Agrobacterium tumefaciens containing binary vector pBin19. Octopine-producing strains have previously been reported to be ineffective in transforming Brassica. Four actively growing yellow/ green sectors selected from on 50 mg/l kanamycin and plants regenerated. Analysis for NPT-II activity these young initially indicated no expression bacterial gene. The nevertheless grown maturity, selfed S1 seed was collected. Three produced microspores which 4 20 times more tolerant than original parent. Southern analysis revealed that one plant (EC-1) had a single site insertion other two (EC-2 EC-6) sites with sequence homology Microspores EC-2 EC-6 transgenics approximately five level tolerated by untransformed plants, while EC-1 transgenic tolerance kanamycin. progeny 100% exhibited trait through both expressing microspore-derived embryos.
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