Regeneration and microprojectile -mediated ransformation in Vida faba L. [electronic resource].

Includes references.

Ten Egyptian cultivars of faba bean (Vicia faba L.) were evaluated for their regeneration capability. Regeneration frequency varied among the cultivars from 30-92% when using embryo axes, while the shoot apex explants revealed lower regeneration frequency (12-57%). Two cultivars (Giza 716 and Giza 843) exhibited no regeneration when using the shoot apex explants. Explants were cultured on Murashige and Skoog (MS) basal salts, with 1 mg/l B5 vitamins, 4.5 mg/l BAP and 0.5 mg/l NAA for mature embryo explants, or 5 mg/l BAP and 1 mg/l NAA for shoot apex explants. The cultures were incubated for one week in the dark during the shoot induction stage to eliminate the browning caused by the accumulation of phenolic compounds. Addition of activated charcoal to the regeneration medium showed negative impact on the regeneration frequency. In vitro grafting method was successfully used for all tested faba bean cultivars. Transformation using microprojectile bombardment protocol was developed using the mature embryo as the explant. The plasmid pCGP1258, harboring the herbicide resistant gene (bar) as a selectable marker and the reporter β- glucuronidase gene (gus), was used for adapting transformation in faba bean cultivars. Mannitol at a concentration of 0.4 M, 1100-psi pressure and 6 cm shooting distance were the most efficient transformation condition. Integration and expression of the transgenes were confirmed by Southern blotting, PCR and histochemical GUS assay. The transformation frequency was as high as 2% for the cultivar Giza 40.

Summary in Arabic.

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