Abstract
Plant transformation via Agrobacterium can be limited by both host specificity and the inability of Agrobacterium to reach the proper cells in the target tissue. Described here is a new and efficient Agrobacterium-based transformation technology that overcomes these barriers and enhances DNA transfer in such diverse plant groups as dicots, monocots, and gymnosperms. This new technology, called sonication-assisted Agrobacterium-mediated transformation (SAAT), involves subjecting the plant tissue to brief periods of ultrasound in the presence of Agrobacterium. Scanning electron and light microscopy reveal that SAAT treatment produces small and uniform fissures and channels throughout the tissue allowing the Agrobacterium easy access to internal plant tissues. Unlike other transformation methods, this system has the potential to transform meristematic tissue buried under several cell layers. SAAT increases transient transformation efficiency in several different plant tissues including leaf tissue, immature cotyledons, somatic and zygotic embryos, roots, stems, shoot apices, embryogenic suspension cells and whole seedlings. A 100- to 1400-fold increase in transient β- glucuronid ase expression has been demonstrated in various tissues of soybean, Ohio buckeye, cowpea, white spruce, wheat and maize. Stable transformation of both soybean and Ohio buckeye has been obtained using SAAT of embryogenic suspension culture tissues. For soybean, SAAT treatment was necessary to obtain stable transformation with this tissue
Similar content being viewed by others
References
Bechtold, N., Ellis, J. and Pelletier, G. (1993) In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. C. R. Acad. Sci. Paris, Life Sciences 316, 1194–9.
Becker, D. (1990) Binary vectors which allow the exchange of plant selectable markers and reporter genes. Nucl. Acid Res. 18, 203.
Bidney, D., Scelonge, C., Martich, J., Burrus, M., Sims, L. and Huffman, G. (1992) Microprojectile bombardment of plant tissues increases transformation frequency by Agrobacterium tumefaciens. Plant Mol. Biol. 18, 301–13.
Chee, P.P., Fober, K.A. and Slightom, J.L. (1989) Transformation of soybean (Glycine max) by infecting germinating seeds with Agrobacterium tumefacients. Plant Physiol. 91, 1212–8.
Finer, J.J. and McMullen, M.D. (1991) Transformation of soybean via particle bombardment of embryogenic suspension culture tissue. In Vitro Cell and Develop. Biol.-Plant 27P, 175–82.
Finer, J.J. and Nagasawa, A. (1988) Development of an embryogenic suspension culture of soybean [Glycine max (L.) Merrill]. Plant Cell Tiss. Org. Cult. 15, 125–36.
Finer, J.J., Finer, K.R. and Santarem, E.R. (1996) Plant cell transformation, physical methods for. In Meyers, R.A. ed., Encyclopedia of Molecular Biology and Molecular Medicine, Weinheim; Germany: VCH Publishers, pp. 458–65.
Frizzel, L.A. (1988) Biological effects of acoustical cavitation. In Suslick, K. ed. Ultrasound, its Chemical, Physical, and Biological Effects Weinheim; Germany: VCH Publishers, pp. 287–303.
Gamborg, O.L., Miller, R.A. and Ojima, K. (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50, 151–8.
Hadi, M.Z., McMullen, M.D. and Finer, J.J. (1996) Transformation of 12 different plasmids into soybean via particle bombardment. Plant Cell Reports 15, 500–5.
Hiei, Y., Ohta, S., Komari, T. and Kumashiro, T. (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J. 6, 271–82.
Hood, E.E., Gelvin, S.B., Melchers, L.S. and Hoekema, A. (1993) New Agrobacterium helper plasmids for gene transfer to plants. Transgenic Res. 2, 208–18.
Horsch, R.B., Fry, J.E., Hoffman, N.L., Eicholtz, D., Rogers, S.G. and Fraley, R.T. (1985) A simple and general method for transferring genes into plants. Science 227, 1229–31.
Ishida, Y., Saito, H., Ohta, S., Hiei, Y., Komari, T. and Kumashiro, T. (1996) High efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens. Nature Biotechnol. 14, 745–50.
Jefferson, R.A. (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5, 387–405.
Joersbo, M. and Brunstedt, J. (1990). Direct gene transfer to plant protoplasts by mild sonication. Plant Cell Rep. 9, 207–10.
Joersbo, M. and Brunstedt, J. (1992) Sonication: A new method for gene transfer to plants. Physiol. Plant. 85, 230–4.
Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–97.
Perl, A., Lotan, O., Abu-Abied, M. and Holland, D. (1996) Establishment of an Agrobacterium-mediated transformation system for grape (Vitis vinifera L.): The role of antioxidants during grape-Agrobacterium interactions. Nature Biotechnol. 14, 624–8.
Saghai-Maroof, M.A., Saliman, K.M., Jorgensen, R.A. and Wallard, R. (1984) Ribosomal DNA spacer length polymorphism in barley: Mendelian inheritance; chromosomal location and population dynamics. Proc. Natl Acad. Sci. USA 81, 8014–8.
Santarem, E.R., Pelissier, B. and Finer, J.J. (1997) Effect of explant orientation, pH, solidifying agent and wounding on initiation of soybean somatic embryos. In Vitro Cell Dev. Biol.-Plant 33, 13–19.
Stachel, S.E., Messens, E., Van Montagu, M. and Zambryski, P. (1985) Identification of the signal molecules produced by wounded plant cells which activate the T-DNA transfer process in Agrobacterium tumefaciens. Nature 318, 624–9.
Suslick, K. (1988) Homogenous sonochemistry. In Suslick, K. ed., Ultrasound, its Chemical, Physical, and Biological Effects Weinheim, Germany: VCH Publishers, pp. 123–61.
Tinland, B. and Hohn, B. (1995) Recombination between prokaryotic and eukaryotic DNA: Integration of Agrobacterium tumefaciens T-DNA into the plant genome. In Setlow, J.K. ed., Genetic Engineering Vol. 17, New York USA: Plenum Press, pp. 209–29.
Vancanneyt, G., Schmidt, R., O'Connor-Sanchez, A., Willmitzer, L. and Rocha-Sosa, M. (1990) Construction of an introncontaining 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, 245–50.
Zhang, L., Cheng, L., Xu, N., Zhao, N., Li, C. Yuan, J. and Jia, S. (1991) Efficient transformation of tobacco by ultrasonication. Bio/Technology 9, 996–7.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Trick, H.N., Finer, J.J. SAAT: sonication-assisted Agrobacterium-mediated transformation. Transgenic Res 6, 329–336 (1997). https://doi.org/10.1023/A:1018470930944
Issue Date:
DOI: https://doi.org/10.1023/A:1018470930944