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Acetosyringone promotes high efficiency transformation of Arabidopsis thaliana explants by Agrobacterium tumefaciens

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Abstract

High frequency transformation of Arabidopsis thaliana leaf explants has been obtained using a disarmed Ti plasmid containing the coding region of a neomycin phosphotransferase gene (NPT II) as a selectable marker. The rate of transformation ranged from 55 to 63 percent when acetosyringone (AS), a natural wound response molecule, was added to an Agrobacterium tumefaciens culture prior to incubation with leaf segments. Without acetosyringone, the transformation rate was approximately 2 to 3 percent. Calli resistant to G418 were regenerated into mature flowering plants in the presence of 10 μg/ml G418. Southern analysis and neomycin phosphotransferase assays confirmed the insertion and expression of the NPT II gene in regenerated Arabidopsis plants.

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References

  1. AertsM, JacobsM, HernalsteensJP, VanMontaguM, SchellJ: Induction and in vitro culture of Arabidopsis thaliana crown gall tumours. Plant Sci Lett 17: 43–50, 1979.

    Google Scholar 

  2. AnG, WatsonBD, ChiangCC: Transformation of tobacco, potato, and Arabidopsis thaliana using a binary Ti vector system. Plant Physiol 81: 301–305, 1986.

    Google Scholar 

  3. BevanMW, FlavellRB, ChiltonM-D: A chimaeric antibiotic resistance gene as a selectable marker for plant cell transformation. Nature 304: 184–187, 1983.

    Google Scholar 

  4. ChristiansonML, WarnickDA: Temporal requirement for phytohormone balance in the control of organogenesis in vitro. Dev Biol 112: 494–497, 1985.

    Google Scholar 

  5. EstelleMA, SommervilleCR: The mutants of Arabidopsis. Trends in Genetics 2: 89–93, 1986.

    Google Scholar 

  6. Feldmann K, Marks MD: Rapid and efficient regeneration of plants from explants of Arabidopsis thaliana. Plant Sci (in press).

  7. FraleyRB, RogersSG, HorschRB, SandersPR, FlickJS, AdamsSP, BittnerML, BrandLA, FinkCL, FryJS, GallupiGR, GoldbergSB, HoffmanNL, WooSC: Expression of bacterial genes in plant cells. Proc Natl Acad Sci USA 80: 4803–4807, 1983.

    Google Scholar 

  8. Herrera-EstrellaL, DeBlockM, MessensE, HernalsteensJ-P, VanMontaguM, SchellJ: Chimeric genes as dominant selectable markers in plant cells. EMBO J 2: 987–995, 1983.

    Google Scholar 

  9. HorschRB, FryJE, HoffmanNL, EichholtzD, RogersSG, FraleyRT: A simple and general method for transferring genes into plants. Science 227: 1229–1231, 1984.

    Google Scholar 

  10. Koukolikova-NicolaZ, ShillitoRD, HohnB, WangK, VanMontaguM, ZambryskiP. Involvement of circular intermediates in the transfer of T-DNA from Agrobacterium tumefaciens to plant cells. Nature 313: 191–196, 1985.

    Google Scholar 

  11. MaligaP, Sz-BreznovitisA, MartonL: Streptomycin-resistant plants from callus culture of haploid tobacco. Nature New Biol 244: 29–30, 1973.

    Google Scholar 

  12. ManiatisT, FritschEF, SambrookJ: In: Molecular Cloning. Cold Spring Harbor Press, Cold Spring Harbor, New York, 1982.

    Google Scholar 

  13. MeyerowitzEM, PruittRE: Arabidopsis thaliana and plant molecular genetics. Science 229: 1214–1218, 1985.

    Google Scholar 

  14. MillerJ: Experiments in Molecular Genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1972.

    Google Scholar 

  15. MurashigeT, SkoogF: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Pl 15: 473–497, 1962.

    Google Scholar 

  16. NorthG: A plant joins the pantheon at last? Nature 315: 366–367, 1985.

    Google Scholar 

  17. PruittRE, MeyerowitzEM: Characterization of the genome of Arabidopsis thaliana. J Mol Biol 187: 169–183, 1986.

    Google Scholar 

  18. ReissB, SprengelR, WillH, SchallerH: A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30: 217–223, 1984.

    Google Scholar 

  19. SommervilleCR, McCountP, CasparT, EstelleM, KeithK: Arabidopsis thaliana as a model system for plant genetics and molecular biology. In: FreelingM (ed) Plant Genetics. UCLA Symposia on Molecular and Cellular Biology, Vol 35, Alan R. Liss, New York, 1985, pp 651–660.

    Google Scholar 

  20. StachelSE, MessensE, VanMontaguM, ZambryskiP: Identification of the signal molecules produced by wounded plant cells that activate T-DNA transfer in Agrobacterium tumefaciens. Nature 318: 624–629, 1985.

    Google Scholar 

  21. StachelS, NesterEW, ZambryskiP. A plant cell factor induces Agrobacterium tumefaciens Vir gene expression. Proc Natl Acad Sci USA 83: 379–383, 1986.

    Google Scholar 

  22. StachelS, TimmermanB, ZambryskiP: Generation of single-stranded T-DNA molecules during the initial stages of T-DNA transfer from Agrobacterium tumefaciens to plant cells. Nature 322: 706–712, 1986.

    Google Scholar 

  23. VeltenJ, VeltenL, HainR, SchellJ: Isolation of a dual plant promoter fragment from the Ti plasmid of Agrobacterium tumefaciens. EMBO J 12: 2723–2730, 1984.

    Google Scholar 

  24. WangK, Herrera-EstrellaL, VanMontaguM, ZambryskiP: Right 25 bp terminus sequence of the nopaline T-DNA is essential for and determines direction of DNA transfer from Agrobacterium to the plant genome. Cell 38: 455–462, 1984.

    Google Scholar 

  25. WeeksDP, BeermanN, GriffithOM: A small-scale five-hour procedure for isolating multiple samples of CsCl-purified DNA: Application to isolations from mammalian, insect, higher plant, alga, yeast, and bacterial sources. Anal Biochem 152: 376–385, 1986.

    Google Scholar 

  26. Zambryski P, Herrera-Estrella L, De Block M, Van Montagu M, Schell J: In: Hollaender A, Setlow J (eds) Genetic Engineering, Principles and Methods, Vol VI. Plenum Press.

  27. ZambryskiP, JoosH, GenetelloC, LeemansJ, VanMontaguM, SchellJ: Ti plasmid vector for the introduction of DNA plant cells without alteration of their normal regeneration capacity. EMBO J 2: 2143–2150, 1983.

    Google Scholar 

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Sheikholeslam, S.N., Weeks, D.P. Acetosyringone promotes high efficiency transformation of Arabidopsis thaliana explants by Agrobacterium tumefaciens . Plant Mol Biol 8, 291–298 (1987). https://doi.org/10.1007/BF00021308

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  • DOI: https://doi.org/10.1007/BF00021308

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