Transgenic muskmelon (Cucumis melo L.) plants were produced efficiently by inoculating cotyledon explants with Agrobacterium tumefaciens strain LBA4404 bearing a Ti plasmid with the NPT II gene for kanaymcin resistance. After co-cultivation for three days, expiants were transferred to melon regeneration medium with kanamycin to select for transformed tissue. Shoot regeneration occurred within 3–5 weeks; excised shoots were rooted on medium containing kanamycin before transferring to soil. Morphologically normal plants were produced in three months. Southern blot analysis confirmed that ca. 85% of the regenerated plants contained the NPT gene. Dot blot analysis and leaf callus assay of progeny of transgenic plants verified transmission of the introduced gene(s) to the next generation. Factors affecting transformation efficiency are discussed.
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indole 3 acetic acid
- NPT II:
neomycin phosphotransferase II
Catlin D, Ochoa O, McCormick S, Quiros CF (1988) Celery transformation by Agrobacterium tumefaciens: Cytological and genetic analysis of transgenic plants. Plant Cell Reports 7:100–103
Chilton MD, Currier T, Ferrand K, Bendich J, Gordon M, Nester E (1974) Agrobacterium tumefaciens DNA and PS8 bacteriophage DNA not detected in crown gall tumors. Proc Natl Acad Sci USA 71:3672–3676
Chyi YS, Jorgensen AJ, Goldstein D, Tanksley SD, Loaiza-Figueroa (1986) Locations and stability of Agrobacterium-mediated T-DNA insertions in the Lycopersicon genome. Mol Gen Genet 204:64–69
Dellaporta SL, Wood J, Hicks JB (1985) Maize DNA miniprep. In Molecular Biology of Plants. Malmberg, Messing, Sussex (eds.) p. 36–37
Dirks, R, Van Buggenum M (1989) In vitro plant regeneration from leaf and cotyledon expiants of Cucumis melo L. Plant Cell. Rep. 7:626–627
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Gasser CS, Fraley RT (1989) Genetically engineering plants for crop improvement. Science 244:1293–1299
Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1331
James DJ, Passey AJ, Barbara DJ, Bevan M (1989) Genetic transformation of apple (Malus pumila Mill.) using a disarmed Ti-binary vector. Plant Cell Reports 7:658–661
Maniatis T, Fritsch EF, Sambrook J (1982) “Molecular Cloning: A Laboratory Manual” Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Michelmore R, Marsh E, Seely S, Landry B (1987) Transformation of lettuce (Lactuca sauva) mediated by Agrobacterium tumefaciens. Plant Cell Reports 6:439–442
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–498
Neidz RP, Smith SS, Dunbar KB, Stephens CT, Murakishi HH (1989) Factors influencing shoot regeneration from cotyledonary expiants of Cucumis melo. Plant Cell Tissue and Org Cult 18:313–319
Orts MC, Garcia-Sogo B, Roche MV, Roig LA, Moreno V (1987) Morphpgenetic response of calli derived from primary explants of diverse cultivars of melon. HortSci 22:666
Rogers SG, Horsch RB, Fraley RT (1986) Gene transfer in plants: production of transformed plants using Ti plasmid vectors. Methods in Enzymology 118:627–640
Rothstein SJ, Lahners KM, Lotstein RJ, Carozi NB, Jayne SM, Rice DA (1987) Promoter cassettes, antibiotic-resistance genes, and vectors for plant transformation. Gene 53:153–161
Trulson AJ, Simpson RB, Shahin EA (1986) Transformation of cucumber (Cucumis sativa L.) plants with Agrobacterium rhizogenes. Theor Appl Genet 73:11–15
Communicated by C. Quiros
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Fang, G., Grumet, R. Agrobacterium tumefaciens mediated transformation and regeneration of muskmelon plants. Plant Cell Reports 9, 160–164 (1990). https://doi.org/10.1007/BF00232095
- Transgenic Plant
- Southern Blot Analysis
- Transformation Efficiency