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Cotyledon explants of cucumber (Cucumis sativus L. CV. Poinsett 76) seedlings were cocultivated with disarmed Agrobacterium strain C58Z707 that contained the binary vector plasmid pGA482GG/cpCMV19. The T-DNA region of this binary vector contains plant-expressible genes for neomycin phosphotransferase II (NPT II), β -glucuronidase (GUS), and the coat protein of cucumber mosaic virus strain C (CMV-C). After infection, the cotyledons were placed on Murashige and Skoog medium containing 100 mg kanamycidlite r. Putative transformed embryogenic calli were obtained, followed by the development of mature embryos and their germination to plants. All transformed RO cucumber plants appeared morphologically normal and tested positive for NPT IL Southern blot analysis of selected cucumber DNAs indicated that NPT II, GUS, and CMV-C coat protein genes were integrated into the genomes. Enzyme-linked immunosorbent assay and Western blot analysis indicated that the CMV-C coat protein is present in the protein extracts of progeny plants. These results show that the Agrobacterium-mediated gene transfer system and regeneration via somatic embryogenesis is an effective method for producing transgenic plants in Cucurbitaceae . Many species of the Cucurbitaceae are susceptible to infection by Agrobacterium pathogens (Anderson and Moore, 1979; Smarrelli et al., 1986; Trulson et al., 1986). In addition, pro- cedures for regenerating several of these species have already been established (Chee, 1990a, 1991a, 1991b). The combina- tion of these two facts suggests that the transfer of genetic in- formation into the genome of cucurbit species via an Agrobacterium -mediated transfer system should be feasible (Chee, 1990b). Such a transfer system would be useful for engineered genes that confer protection from viral infection (Cuozzo et al., 1988; Powell-Abel et al., 1986) and herbicides (Comai et al., 1985; dells-Cioppa et al., 1987; Lee et al, 1988; Stalker et al., 1985). Most of these genetic traits are difficult to acquire via conventional plant breeding, even virus protection (Providenti, 1985). The sequence of the coat protein gene from CMV-C has been reported (Quemada et al., 1989) and has recently been engi- neered for expression in plants (Quemada et al., 1991). The development of cucumber plants that use the coat-protein-me- diated mechanism for CMV protection (Powell-Abel et al., 1986) would be of scientific interest because this would allow for side- by-side comparisons of these genetically engineered plants with those plants that possess naturally occurring resistance. Coat- protein-mediated protection from CMV infections has been demonstrated in tobacco (Cuozzo et al., 1988; Quemada et al., 1991). The present report describes the use of a disarmed strain of Agrobacterium tumefaciens to mediate the transfer of the en- gineered CMV-C coat protein into cucumber. In addition, it describes the use of regeneration via somatic embryogenesis to obtain morphologically normal and fertile transformed plants that express the CMV-C coat protein gene.

Transfer and Expression of Cucumber Mosaic Virus Coat Protein Gene in the Genome of Cucumis sativus