Abstract
The protocol detailed below describes an efficient method for Agrobacterium tumefaciens-mediated transformation of Brassica napus. cv Westar based on two earlier published procedures (Radke et al. 1988; De Block et al. 1989). Hypocotyl explants are cocultivated in liquid medium with the nopaline-type Agrobacterium strain MOG301 carrying different binary vectors comprising the gus-intron reporter gene and a selectable marker. Transgenic plants are produced with kanamycin, hygromycin B, phosphinothricin, or chlorsulfuron as selective agents.
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References
Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucl Acids Res 12: 8711–8721
De Block M, Debrouwer D, Tenning P (1989) Transformation of Brassica napus and Brassica oleracea using Agrobacterium tumefaciens and the expression of the bar and neo genes in the transgenic plants. Plant Physiol 91: 694–701
Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50: 151–158
Haughn GW, Smith J, Mazur B, Somerville C (1988) Transformation with a mutant Arabidopsis acetolactate synthase gene renders tobacco resistant to sulfonylurea herbicides. Mol Gen Genet 211: 266–271
Hood EE, Gelvin SB, Melchers LS, Hoekema A (1993) New Agrobacterium helper plasmid for gene transfer to plants. Transgenic Res 2: 208–218
Jefferson RA (1987) Assaying chimaeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5 (4): 387–405
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15: 473–497
Radke SE, Andrews BM, Moloney MM, Crouch ML, Kridl JC, Knauf VC (1988) Transformation of Brassica nap us L. using Agrobacterium tumefaciens: developmentally regulated expression of a reintroduced napin gene. Theor Appl Genet 75: 685–694
Vancanneyt G, Schmidt R, O’Connor-Sanchez A, Willmitzer L, Rocha-Sosa M (1990) Construction of an intron-containing marker gene: Splicing of the intron in transgenic plants and its use in monitoring early events in Agrobacterium-mediated plant transformation. Mol Gen Genet 220: 245–250
Van den Elzen PJM, Townsend J, Lee KY, Bedbrook JR (1985) A chimaeric hygromycin resistance gene as selectable marker in plant cells. Plant Mol Biol 5: 299–302
Wohlleben W, Arnold W, Broer I, Hilleman D, Strauch E, Pühler A (1988) Nucleotide sequence of the phosphinothricin N-acetyltransferase gene from from Streptomyces viridochromogenes Tä494 and its expression in Nicotiana tabacum. Gene 70: 25–37
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© 1995 Springer-Verlag Berlin Heidelberg
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Bade, J.B., Damm, B. (1995). Introduction II Agrobacterium-rhizogenes,a Natural Transformation System. In: Potrykus, I., Spangenberg, G. (eds) Gene Transfer to Plants. Springer Lab Manual. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-79247-2_5
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DOI: https://doi.org/10.1007/978-3-642-79247-2_5
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-48967-9
Online ISBN: 978-3-642-79247-2
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