Abstract: To establish the efficient Agrobacterium-mediated transformation system of pepper (Capsicum annuum L.)Flamingo-bill explant, the binary expression vector containing Cry2Aa2 and PamPAP was transferred into pepper mediatedby Agrobacterium tumefaciens and the transformation system was optimized. The results showed that adventitiousdifferential rates of Flamingo-bill was significantly higher than that of cotyledon with petiole in three pepper cultivars, and the adventitious shoots of Flamingo-bill were easier to elongate. The highest adventitious differential rate and resistant rate were obtained when the Flamingo -bill explants were selected at 4 ~6 days later after cocultivation. The optimal concentration for screening the transformed Flamingo-bill explants was 18 g/L mannose and 12 g/L sucrose. The optimal rooting medium for resistant seedlings was MS+NAA 0.2 mg/L+IAA 0.1 mg/L. 28 transgenic pepper plants were obtained with optimizated PMI/Man selection system, of which the positive and transformation rate were 21.4% and 2.5% , respectively.