Abstract
Adventitious shoots formed on the proximal cut edges of different cotyledonary explants of watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai; cvs. Sweet Gem and Gold Medal] cultured on Murashige and Skoog's (MS) medium with 1 mgl-1 6-benzyladenine (BA). Light (16-h photoperiod, about 7 Wm-2 cool-white fluorescent lamps) was essential for shoot formation. To obtain transformed plants, cotyledonary explants of ‘Sweet Gem’ were cocultured with Agrobacterium tumefaciens LBA4404, a disarmed strain harboring a binary vector pBI121 carrying the CaMV 35S promoter-β-glucuronidase (GUS) gene fusion used as a reporter gene and NOS promoter-neomycin phosphotransferase gene as a positive selection marker, for 48 h on MS medium with 1 mgl-1 BA and 200 μM β-hydroxyacetosyringone. After 48 h of culture, explants were transferred to medium with 1 mgl-1 BA 250 mgl-1 carbenicillin, and 100 mgl-1 kanamycin and cultured in the light. Adventitious shoots formed on the explants after 4 weeks of culture. When subjected to GUS histochemical assay, young leaves obtained from the shoots showed a positive response at a frequency of up to 16%. Preculturing cotyledonary explants on MS medium with 1 mgl-1 BA for 5 d enhanced the competence of the cells to be transformed by Agrobacterium. Southern blot analysis confirmed that the GUS gene was incorporated into the genomic DNA of the GUS-positive regenerants. The transformed plants were grown to maturity.
Similar content being viewed by others
References
Anghel I, Rosu A (1985) Rev Roum Biol-Biol Végét 30:43–55
Broglie K, Chet I, Hollyday M, Cressman R, Biddle P, Knowltion S, Mauvais CJ, Brgolie R (1991) Science 254:1194–1197
Compton ME, Gray DJ (1993a) J Amer Soc Hort Sci 118:151–157
Compton ME, Gray DJ (1993b) Plant Cell Rep 12:61–65
Deblaere R, Reynaerts A, Hofte H, Hernalsteens JP, Leemans J, Van Montagu M (1987) Methods in Enzymol 153:277–292
Dong J-Z, Jia S-R (1991) Plant Cell Rep 9:559–562
Jefferson RA, Kavanaugh TA, Bevan MW (1987) EMBO J 6:3901–3907
Müller-Röber B, Sonnewald U, Willmitzer L (1992) EMBO J 11:1229–1238
Murashige T, Skoog F (1962) Physiol Plant 15:156–163
Srivastava DR, Andrianov VM, Piruzian ES (1989) Plant Cell Rep 8:300–302
Author information
Authors and Affiliations
Additional information
Communicated by G. C. Phillips
Rights and permissions
About this article
Cite this article
Choi, P.S., Soh, W.Y., Kim, Y.S. et al. Genetic transformation and plant regeneration of watermelon using Agrobacterium tumefaciens . Plant Cell Reports 13, 344–348 (1994). https://doi.org/10.1007/BF00232634
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00232634