Agrobacterium-mediated transformation of cucumber (Cucumis sativus L.) using a sense mitogen-activated protein kinase gene (CsNMAPK)

The objective of this study was to develop an efficient regeneration and transformation protocol for cucumber (Cucumis sativus L.). Cotyledonary node explants showed a very high regeneration frequency (94.3 %) when incubated on Murashige and Skoog medium (MS) supplemented with 1.5 mg l−1 6-benzyladenine (BA). An Agrobacterium-mediated transformation methodology for cucumber was developed using this protocol. A construct containing a sense mitogen-activated protein kinase gene (CsNMAPK) was used to transform cucumber cotyledonary nodes. Adventitious shoots were obtained in selection media containing 150 mg l−1 kanamycin and 250 mg l−1 carbenicillin 4 weeks after bacterial inoculation. A total of twelve transgenic plants were confirmed using PCR-specific analysis. The transgene copy number of four transgenic plants (random selection) was found to be a single gene copy using Southern blot analysis and the expression level of the CsNMAPK gene in the transgenic plants was significantly higher than in the wild-type plants. The transgenic rate was approximately 4.8 %.