Summary
A method for Agrobacterium-mediated transformation of Citrus and organogenic regeneration of transgenic plants is reported. Internodal stem segments were co-cultured with Agrobacterium harboring binary vectors that contained the genes for the scorable marker ß-glucuronidase (GUS) and the selectable marker NPT-II. A low but significant percentage (≤ 5%) of the shoots regenerated in the presence of 100 μg/ml kanamycin were GUS+. Polymerase chain reaction (PCR) analysis confirmed that GUS+ shoots contained T-DNA. Two plants established in soil were shown to be transgenic by Southern analysis.
Similar content being viewed by others
References
An G (1987) Methods Enzymol 153:292–305
Watson BD, Stachel S, Gordon MP, Nester EW (1985) EMBO J 4:277–284
Church GM, Gilbert W (1984) Proc Natl Acad Sci 81:1991–1995
Dellaporta SL, Wood J, Hicks JB (1983) Plant Mol Biol Reporter 1:19–21
Durham RE (1990) PhD Dissertation, University of Florida
Gmitter FG, Moore GA (1986) Cell Tissue and Organ Culture 6:139–147
Hidaka T, Omura M, Ugaki M, Tomiyama M, Kato A, Ohshima M, Motoyoshi F (1990) Japan J Breed 40:199–207
Hood EE, Helmer GL, Fraley RT, Chilton M-D (1986) J Bacteriol 168:1291–1301
Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) Science 227:1229–1231
Jayaswal RK, Veluthambi K, Gelvin SB, Slightom, JL (1987) J Bacteriol 169:5035–5045
Jefferson RA (1987) Plant Mol Biol Reporter 5:387–405
Jordan MC, McHughen A (1988) Plant Cell Rep 7:285–287
Knbayashi S, Uchimiya H (1989) Japan J Genet 64:91–97
Komari T, Halperin W, Nester EW (1986) J Bacteriol 166:88–94
Lassner MW, Peterson P, Yoder JI (1989) Plant Mol Biol Reporter 7:116–128
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, New York.
Murashige T, Tucker DPH (1969) Proc First Inti Citrus Symp 3:1155–1161
Rogers, SG, Klee HJ, Horsch RB, Fraley RT (1987) Meth Enzymol 153:253–277
Rogers SO, Bendich AJ (1985) Plant Mol Biol 5:69–76
Vardi A, Bleichman S, Aviv D (1990) Plant Sci 69:199–206
Author information
Authors and Affiliations
Additional information
Communicated by G. C. Phillips
Rights and permissions
About this article
Cite this article
Moore, G.A., Jacono, C.C., Neidigh, J.L. et al. Agrobacterium-mediated transformation of Citrus stem segments and regeneration of transgenic plants. Plant Cell Reports 11, 238–242 (1992). https://doi.org/10.1007/BF00235073
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00235073