Agrobacterium-mediated transformation of Citrus stem segments and regeneration of transgenic plants

Plant Cell Reports - Tập 11 - Trang 238-242 - 1992
G. A. Moore1, C. C. Jacono1, J. L. Neidigh1, S. D. Lawrence1, K. Cline1
1Fruit Crops Department, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, USA

Tóm tắt

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.

Tài liệu tham khảo

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