Summary
Somatic embryos have been successfully used as a target tissue for transformation and regeneration of transgenic walnut plants. Walnut somatic embryos, initiated originally from developing zygotic embryos, proliferate numerous secondary embryos from single cells in the epidermal layer. These single cells in intact somatic embryos are susceptible to transformation by genetically engineeredAgrobacterium tumefaciens and provide a means to regenerate nonchimeric transgenic plants. This gene transfer system has been made more efficient using, a) vector plasmids containing two marker genes encoding β-glucuronidase (GUS) and aminoglycoside phosphotransferase (APH(3′)II) and B) a more virulent strain ofAgrobacterium. This system should be applicable to any crop that undergoes repetitive embryogenesis from singleAgrobacterium-susceptible cells.
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References
Ammirato, P. V. Organizational events during somatic embryogenesis. In: Green, C. E.; Somers, D. A.; Hackett, W. P., et al. eds. Plant tissue and cell culture. New York: Alan R. Liss; 1987:57–81.
Ammirato, P. V. Embryogenesis. In: Evans, D. A.; Sharp, W. R.; Ammirato, P. V., et al., editors. Handbook of plant cell culture, vol. 1. New York: Macmillan; 1983:82–123
Ausubel, F. M.; Brent, R.; Kingston, R. E., et al. Current protocols in molecular biology, vols. 1 & 2. New York: Greene Publishing Associates and Wiley-Interscience; 1987.
Barker, R. F.; Idler, K. B.; Thompson, D. V., et al. Nucleotide sequence of the T-DNA region from theAgrobacterium tumefaciens octopine Ti plasmid pTil5955. Plant Mol. Biol. 2:335–350; 1983.
Beck, E.; Ludwig, G.; Auerswald, E. A., et al. Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene 19:327–336; 1983.
Dandekar, A. M.; Martin, L. A.; McGranahan, G. H. Genetic transformation and foreign gene expression in walnut tissue. J. Am. Soc. Hort. Sci. 113:945–949; 1988.
Dandekar, A. M.; Uratsu, S. L.; Matsuta, N.Agrobacterium-mediated transformation of apple: factors influencing virulence. Acta Hort. 1989. (In press).
Dellaporta, S. L.; Wood, J.; Hicks, J. B. A plant DNA minipreparation: version II. Plant Mol. Biol. Rep. 1(2):19; 1983.
Driver, J. A.; Kuniyuki, A. H. In vitro propagation of Paradox walnut rootstock. Hortscience 19:507–509; 1984.
Feinberg, A. P.; Vogelstein, B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132:6–13; 1983.
Feinberg, A. P.; Vogelstein, B. Addendum to a technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 137:266–267; 1984.
Holsters, M.; De Waele, D.; Depicker, A., et al. Transfection and transformation ofAgrobacterium tumefaciens. Mol. Gen. Genet. 163:181–187; 1978.
Hood, E. H.; Helmer, G. L.; Fraley, R. T., et al. The hypervirulence ofAgrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside T-DNA. J. Bacteriol. 168:1291–1301; 1986.
Horsch, R. B.; Fry, J. E.; Hoffman, N. L., et al. A simple and general method for transferring genes into plants. Science 227:1229–1231; 1985.
Jefferson, R. A. Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol. Biol. Rep. 5:387–405; 1987.
Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685; 1970.
Maniatis, T.; Fritsch, E. F.; Sambrook, J. Molecular cloning: a laboratory manual. New York: Cold Spring Harbor Laboratory; 1982.
McGranahan, G. H.; Driver, J. A. Tulecke, W. Tissue culture ofJuglans. In: Bonga, J. M.; Durzan, D. J., eds. Cell and tissue culture in forestry, vol. 3. Boston: Martinus Nijhoff; 1987:261–271.
McGranahan, G. H.; Leslie, C. A.; Uratsu, S. L., et al.Agrobacterium-mediated transformation of walnut somatic embryos and regeneration of transgenic plants. Bio/Technology 6:800–804; 1988.
McGranahan, G. H.; Leslie, C. A.; Uratsu, S. L., et al. Improved efficiency of the walnut somatic embryo gene transfer system. Plant Cell Rep. 1990. (In press).
Polito, V. S.; McGranahan, G. H.; Pinney, K., et al. Origin of somatic embryos from repetitively embryogenic cultures of walnut (Juglans regia L.): implications forAgrobacterium-mediated transformation. Plant Cell Rep. 8:219–221; 1989.
Reiss, B.; Sprengle, R.; Will, H., et al. A new and sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extract. Gene 30:211–218; 1984.
Rodriguez, R. L.; Tait, R. C. Recombinant DNA Techniques. Menlo Park, CA: Benjamin/Cummings; 1983.
Southern, E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98:503–507; 1975.
Tulecke, W.; McGranahan, G. H. Somatic embryogenesis and plant regeneration from cotyledon tissue of walnut,Juglans regia L. Plant Sci. 40:53–67; 1985.
Tulecke, W. Somatic embryogenesis in woody perennials. In: Bonga, J. M.; D. J. Durzan, D. J., eds. Cell and tissue culture in forestry, vol. 2. Boston: Martinus Nijhoff; 1987:61–91.
Wann, S. R. Somatic embryogenesis in woody species. Hort. Rev. 10:153–182; 1988.
Wahl, G. M.; Stern, M.; Stark, R. G. Efficient transfer of large DNA fragments from agarose gels to diazobenzloxymethal paper and rapid hybridization using dextran sulfate. Proc. Natl. Acad. Sci. USA 76:3683–3687; 1977.
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Dandekar, A.M., McGranahan, G.H., Leslie, C.A. et al. Agrobacterium-mediated transformation of somatic embryos as a method for the production of transgenic plants. Journal of Tissue Culture Methods 12, 145–150 (1989). https://doi.org/10.1007/BF01404441
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DOI: https://doi.org/10.1007/BF01404441