Agrobacterium-mediated transformation of Brassica campestris spp. Parachinensis with synthetic Bacillus thuringiensis cry1Ab and cry1Ac genes

An effective plant regeneration procedure and a gene transfer system via Ag robacterium tumefaciens were developed in Brassica campestris ssp. parachin ensis. Hypocotyls from 5-day-old seedlings with 2 days pre-culture were inf ected with Agrobacterium strain MOG301 harboring a binary vector containing a synthetic Bacillus thuringiensis (B.t.) cry1Ab or cry1Ac gene with full codon-modification. After culture and selection on MS medium supplemented w ith 4.0 mg/l BAP, 2.0 mg/l NAA, 70 mu M AgNO3 and 50 mg/l kanamycin, a numb er of kanamycin-resistant plantlets were regenerated. PCR and Southern blot ting analysis were used to identify and characterize the transgenic plants with the integrated cry1Ab or cry1Ac gene. Western blotting analysis of the transgenic plants confirmed the expression of insecticidal proteins encode d by cry1Ab or cry1Ac. Subsequent bioassay with larvae of the Diamondback m oth, Plutella xylostella, demonstrated that the transgenic plants were resi stant to feeding damage.