THE EFFECT OF GROWTH-REGULATORS AND ANTIBIOTICS ON EGGPLANT TRANSFORMATION

A protocol with a high rate of transformation and regeneration of 'Hib ush' 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 cocultivate d with Agrobacterium tumefaciens strains Q10, Q20, Q30, Q40, Q201, Q20 2, Q203, or Q203 containing the native cryIIIB Bacillus thuringiensis (Bt), neomycin phosphotransferase (NPTII), and beta-glucuronidase (uid A) genes, a callus/bud regeneration frequency of 38.8% was observed on the selection medium, Kanamycin at 50 mu g . mL(-1) was most effectiv e in selecting for transgenic buds and shoots, Augmentin at 300 mu . m L(-1) was used to eliminate A. tumefaciens. Augmentin also enhanced sh oot proliferation. A transformation/regeneration efficiency of 20.8% w as observed for shoot production, More than 400 putative transgenic pl ants have been produced with this method. From 50 putative transgenic plants, gene integration has been confirmed with Southern blot analysi s and progeny tests.