The effect of growth regulators and antibiotics on eggplant transformation

A protocol with a high rate of transformation and regeneration of 'Hibush' eggplant (Solanum,melongena L,) has been developed, This protocol used leaves of in vitro-grown seedlings as a source of explants. The shoot regeneration culture medium contained 0.1 mu M thidiazuron (TDZ) combined with 10 to 20 mu M N-6-[isopentyl] adenine (2iP). Adding TDZ significantly improved regeneration efficiency and produced a mean of 15 buds and 3 to 4 shoots per explant, When explants were cocultivated with Agrobacterium tumefaciens strains Q10, Q20, Q30, Q40, Q201, Q202, Q203, or Q203 containing the native cryIIIB Bacillus thuringiensis (Bt), neomycin phosphotransferase (NPTII), and beta-glucuronidase (uidA) genes, a callus/bud regeneration frequency of 38.8% was observed on the selection medium, Kanamycin at 50 mu g . mL(-1) was most effective in selecting for transgenic buds and shoots, Augmentin at 300 mu . mL(-1) was used to eliminate A. tumefaciens. Augmentin also enhanced shoot proliferation. A transformation/regeneration efficiency of 20.8% was observed for shoot production, More than 400 putative transgenic plants have been produced with this method. From 50 putative transgenic plants, gene integration has been confirmed with Southern blot analysis and progeny tests.