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Efficient Transfer of a Glyphosate Tolerance Gene into Tomato Using a Binary Agrobacterium Tumefaciens Vector

Abstract

An efficient transformation system was developed for tomato by studying several factors that affect the rate of Agrobacterium mediated transformation of explant tissue. Cotyledons of tomato were co–cultivated with Agrobacterium tumefaciens harboring a binary vector with two neomycin phosphotransferase (NPT) II genes and a mutant aroA gene. Over 100 transgenic plants were regenerated and rooted on medium containing kanamycin and eighty percent of these plants exhibited both NPT II enzyme activity and produced the mutant aroA protein. Progeny of aroA positive plants were tolerant to glyphosate at concentrations of 0.84 kg active ingredient/ha. The tolerance phenotype segregated in a manner consistent with Mendelian inheritance.

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References

  1. Baird, D.D., Upshiurch, R.P., Homesely, W.B. and Franz, J.E. 1971. Introduction of a new broad spectrum post-emergence herbicide class with utility for herbaceous perennial weed control. Proc. North Cent. Weed Control Conf. 26: 6468.

    Google Scholar 

  2. Barker, R.F., Idler, K.B., Thompson, D.V. and Kemp, J.D. 1983. Nucleotide sequence of the T-DNA region from the Agrobacterium tumefaciens octopine Ti plasmid pTi15955. Plant Mol. Biol. 2: 335–350.

    Article  CAS  Google Scholar 

  3. Bernatzky, R. and Tanksley, S.D. 1986. Towards a saturated linkage map in tomato based on isozymes and random cDNA sequences. Genetics 112: 887–898.

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Bevan, M., Flavell, R.B., and Chilton, M.-D. 1983. A chimeric antibiotic resistance gene as a selectable marker for plant cell transformation. Nature 304: 184–187.

    Article  CAS  Google Scholar 

  5. Comai, L., Sen, L., and Stalker, D. 1983. An altered aroA gene product confers resistance to the herbicide glyphosate. Science 221: 370–371.

    Article  CAS  Google Scholar 

  6. Comai, L., Facciotti, D., Hiatt, W.R., Thompson, G., Rose, R., and Stalker, D. 1985. Expression in plants of a mutant aroA gene from Salmonella typhimurium confers tolerance to glyphosate. Nature 317: 741–744.

    Article  CAS  Google Scholar 

  7. Dellaporta, S.L., Wood, J., and Hicks, J.B. 1983. A plant DNA minipreparation: version II. Plant Mol. Bio. Rep. 1: 19–21.

    Article  CAS  Google Scholar 

  8. Ditta, G., Stanfield, S., Corbin, D., Helinski, D. 1980. Broad host range DNA cloning system for gram-negative bacteria: Construction of a gene bank of Rhizobium meliloti. Proc. Natl. Acad. Sci USA 77: 7347–7351.

    Article  CAS  Google Scholar 

  9. Fillatti, J.J., Sellmer, J., McCown, B., Haissig, B., and Comai, L. 1987. Agrobacterium mediated transformation and regeneration of Populus. Mol. Gen. Genet. 206: 192–199.

    Article  CAS  Google Scholar 

  10. Folmar, L.C., Sanders, H.O., Julin, A.M. 1979. Toxicity of the herbicide glyphosate and several of its formulations to fish and aquatic invertebrates. Arch. Environ. Contam. Toxicol. 8: 269–278.

    Article  CAS  Google Scholar 

  11. Fraley, R.T., Rogers, S.G., Horsch, R.B., Sanders, P., Flick, J., Adams, S., Bittner, M., Brand, L., Fink, C., Fry, J., Galluppi, G., Goldberg, S., Hoffmann, N., and Woo, S. 1983. Expression of bacterial genes in plant cells. Proc. Natl. Acad. Sci. USA 80: 4803–4807.

    Article  CAS  Google Scholar 

  12. Garfinkel. 1980. Agrobacterium tumefaciens mutants affected in crown gall tumorigenesis and octopine catabolism. J. Bacteriol. 144: 732–743.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Herrera-Estrella, L., Depicker, A., Van Montagu, M. and Schell, J. 1983. Expression of chimeric genes transferred into plant cells using Ti plasmid-derived vector. Nature 303: 209–213.

    Article  CAS  Google Scholar 

  14. Hoekema, A., Hirsch, P.R., Hooykaas, P.J.J. and Schilperoort, R.A. 1983. A binary plant vector strategy based on separation of vir and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303: 179–180.

    Article  CAS  Google Scholar 

  15. Horsch, R.B., Fry, J.B., Hoffmann, N.L., Wallroth, M., Eichholtz, D., Rogers, S.G. and Fraley, R.T. 1985. A simple and general method for transferring genes into plants. Science 227: 1229–1231.

    Article  CAS  Google Scholar 

  16. Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

  17. McCormick, S., Niedermeyer, J., Fry, J., Barnason, A., Horsch, K., Fraley, R. 1986. Leaf disc transformation of cultivated tomato (L. esculentum) using Agrobacterium tumefaciens. Plant Cell Reports 5: 81–84.

    Article  CAS  Google Scholar 

  18. Melton, D.A., Krieg, P.A., Rebagliati, M.R., Maniatis, T., Zinn, K. and Green, M.R. 1984. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acid. Res. 12: 7035–7056.

    Article  CAS  Google Scholar 

  19. Reiss, B., Sprengle, R., Will, H. and Schaller, H. 1984. A new sensitive method for qualitative and quantitative assay of neomycin phospho-transferase in crude cell extracts. Gene 30: 211–217.

    Article  CAS  Google Scholar 

  20. Rick, C.M. 1983. Tomato (Lycopersicon) In: Isozymes in Plant Genetics and Breeding. S. D. Tanksley and T.J. Orton (eds.). Elsevier Co., Amsterdam.

    Google Scholar 

  21. Shah, D., Horsch, R., Klee, H., Kishore, G., Winter, J., Tumer, N., Hironaka, C., Sanders, P., Gasser, C., Aykent, S., Siegel, N., Rogers, S. and Fraley, R. 1982. Engineering herbicide tolerance in transgenic plants. Sci. 233: 478–481.

    Article  Google Scholar 

  22. Shahin, E.A., Spielmann, A., Sukhapinda, K., Simpson, R.B. and Yashar, M. 1986. Transformation of cultivated alfalfa using disarmed Agrobacterium tumefaciens. Crop Sci. 26: 1235–1239.

    Article  CAS  Google Scholar 

  23. Sinich, 1982. Hypothesis testing, p. 313–362. In: Statistics by Example. Peller, Santa Clara, CA.

    Google Scholar 

  24. Stachel, S.E., Messens, E., Van Montagu, M. and Zambryski, P. 1985. Identification of the signal molecules produced by wounded plant cells that activate T-DNA transfer in Agrobacterium tumefaciens. Nature 318: 624–629.

    Article  Google Scholar 

  25. Thomas, B.R. and Pratt, D. 1981. Efficient hybridization between Lycopersicon esculentum and L. peruvanum via embryo callus. Theor. Appl. Genet. 59: 215–219.

    Article  CAS  Google Scholar 

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Fillatti, J., Kiser, J., Rose, R. et al. Efficient Transfer of a Glyphosate Tolerance Gene into Tomato Using a Binary Agrobacterium Tumefaciens Vector. Nat Biotechnol 5, 726–730 (1987). https://doi.org/10.1038/nbt0787-726

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  • DOI: https://doi.org/10.1038/nbt0787-726

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