We established improved methods for Agrobacterium-mediated transformation of cucumber (Cucumis sativus L.) and kabocha squash (Cucurbita moschata Duch). Vacuum infiltration of cotyledonary explants with Agrobacterium suspension enhanced the Agrobacterium infection efficiency in the proximal regions of explants. Wounding treatment was also essential for kabocha squash. Cocultivation on filter paper wicks suppressed necrosis of explants, keeping regeneration efficacy. Putative transgenic plants were screened by kanamycin resistance and green fluorescent protein (GFP) fluorescence. These putative transgenic plants grew normally and T1 seeds were obtained, and stable integration and transmission of the transgene in T1 generations were confirmed by Southern hybridization and PCR. The average transgenic efficiency for cucumber and kabocha squash was 11.9 ± 3.5 and 9.2 ± 2.9 %, respectively.
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