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Optimal levels of antibiotics to eliminate Agrobacterium and in vitro selection in guava


, : Optimal levels of antibiotics to eliminate Agrobacterium and in vitro selection in guava. Hortscience 36(3): 535, June

Five concentrations each of the antibiotics hygromycin, kanamycin, cefatoxime and carbenecillin were evaluated to determine minimum inhibitory concentrations for in vitro shoot regeneration from guava (Psidium guajava L.) expants. Young nodes and internodes from two guava genotypes 'Redfleshed' and 'Lucknow-49' grown in the greenhouse in Fort Valley, were employed as explants. The Murashiege and Skoog (MS) basal medium was supplemented separately with hygromycin (0, 2.5, 5, 10, 15 mg/L), cefatoxime (0, 10, 20, 50, 100 mg/L), kanamycin (0, 50, 100, 250, 500 mg/L), and carbenecillin (0, 50, 100, 250, 500 mg/L). The cultures were evaluated after four weeks of incubation at 26 +- 2degreeC with a 16 h light/8 h dark photoperiod. The guava genotypes showed no significant differences in shoot regeneration due to either antibiotics or their concentrations. Although hygromycin at 2.5 mg/L concentration did not kill explants, it inhibited shoot regeneration. Cefatoxime, on the other hand, was not lethal to explants even at 100 mg/L since they responded to shoot regeneration from nodes. Those nodal explants receiving 500 mg/L kanamycin were still green, but produced no shoots from explants. The nodal explants treated with carbenecillin at 500 mg/L successfully produced shoots. Thus, it was concluded that hygromycin at 2.5 mg/L can be used as an effective selection tool while cefatoxime at 100 mg/L can be used to eliminate Agrobacterium from explants after the cocultivation in guava genetic transformation.

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