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Transformation of cotton (Gossypium hirsutum L.) by Agrobacterium tumefaciens and regeneration of transgenic plants

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Abstract

Cotton (Gossypium hirsutum L.) cotyledon tissues have been efficiently transformed and plants have been regenerated. Cotyledon pieces from 12-day-old aseptically germinated seedlings were inoculated with Agrobacterium tumefaciens strains containing avirulent Ti (tumor-inducing) plasmids with a chimeric gene encoding kanamycin resistance. After three days cocultivation, the cotyledon pieces were placed on a callus initiation medium containing kanamycin for selection. High frequencies of transformed kanamycin-resistant calli were produced, more than 80% of which were induced to form somatic embryos. Somatic embryos were germinated, and plants were regenerated and transferred to soil. Transformation was confirmed by opine production, kanamycin resistance, immunoassay, and DNA blot hybridization. This process for producing transgenic cotton plants facilitates transfer of genes of economic importance to cotton.

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References

  1. Fraley R, Rogers S, Horsch R: Genetic transformation in higher plants. CRC Critical Reviews in Plant Science 4: 1–46 (1986).

    Google Scholar 

  2. Horsch RB, Fry JS, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT: A simple and general method for transferring genes into plants. Science 227: 1229–1231 (1985).

    Google Scholar 

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

    Google Scholar 

  4. Shahin EA, Simpson R: Gene transfer system for potato. HortScience 21: 1199–1201 (1986).

    Google Scholar 

  5. Shahin EA, Spielmann A, Sukhapinda K, Simpson R, Yashar M: Transformation of cultivated alfalfa using disarmed Agrobacterium tumefaciens. Crop Science 26: 1235–1239 (1986).

    Google Scholar 

  6. Zhou G-Y, Weng J, Zeng Y, Huang J, Qian S, Liu G: Introduction of exogenous DNA into cotton embryos. Meth Enzymol 101: 433–481 (1983).

    Google Scholar 

  7. Umbeck P, Johnson G, Barton K, Swain W: Genetically transformed cotton (Gossypium hirsutum L.) plants. Bio-Technology 5: 263–266 (1987).

    Google Scholar 

  8. Firoozabady E, DeBoer DL, Maroney MJ: Transformation and regeneration of cotton, Gossypium hirsutum L. Tailoring Genes for Crop Improvements: An Agricultural Perspective. University of California, Davis, Abstracts (1986) p. 24.

    Google Scholar 

  9. Davidonis GH, Hamilton RH: Plant regeneration from callus tissue of Gossypium hirsutum L. Plant Sci Lett 32: 89–93 (1983).

    Google Scholar 

  10. Shoemaker RC, Couche LJ, Galbraith DW: Characterization of somatic embryogenesis and plant regeneration in cotton (Gossypium hirsutum L.). Plant Cell Rep 3: 178–181 (1986).

    Google Scholar 

  11. Gawel NJ, Rao AP, Robacker CD: Somatic embryogenesis from leaf and petiole callus cultures of Gossypium hirsutum. Plant Cell Rep 5: 457–459 (1986).

    Google Scholar 

  12. Trolinde NL, Goodin JR: Somatic embryogenesis and plant-regeneration in cotton (Gossypium hirsutum L). Plant Cell Rep 6: 231–234 (1987).

    Google Scholar 

  13. Firoozabady E, Sutton DW, Merlo DJ: “Micro” Ti plasmid vectors in plant transformation: open reading frame 1 promoter of pTi15955 TL-DNA directs expression of kanamycin resistance in transformed plant cells (to be submitted to Plant Physiol).

  14. Casadaban MJ, Cohen SN: Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol 138: 179–207 (1980).

    Google Scholar 

  15. Wood WB: Host specificity of DNA produced by Escherichia coli: Bacterial mutations affected the restriction and modification of DNA. J Mol Biol 16: 118–133 (1966).

    Google Scholar 

  16. Ruvkun GB, Ausubel FM: A general method for site-directed mutagenesis in prokaryotes. Nature 289: 85–88 (1981).

    Google Scholar 

  17. Garfinkel DJ, Simpson RB, Ream LW, White FF, Gordon MP, Nester EW: Genetic analysis of crown gall: Fine structure map of the T-DNA by site-directed mutagenesis. Cell 27: 143–153 (1981).

    Google Scholar 

  18. Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA: A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303: 179–180 (1983).

    Google Scholar 

  19. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Press, New York (1982).

    Google Scholar 

  20. Klapwijk PM, deFonge AJR, Schilperoort RA, Rörsch A: An enrichment technique for auxotrophs of Agrobacterium tumefaciens using a combination of carbenicillin and lysozyme. J Gen Microbiol 91: 177–182 (1975).

    Google Scholar 

  21. Ditta G, Schmidhauser T, Yakobson E, Lu P, Liang X-W, Finlay DR, Guiney D, Helinski DR: Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid 13: 149–153 (1985).

    Google Scholar 

  22. Sutton DW, Merlo DJ, Talbot DR: TL-based sub-T-DNA plasmids. European Patent Publication 0, 223, 417: May 27 (1987).

    Google Scholar 

  23. Kemp JD, Sutton DW, Fink C, Barker RF, Hall TC: Agrobacterium mediated transfer of foreign genes into plants. In: Owens LD (ed.) Genetic Engineering: Applications to Agriculture. Rowman and Allanheld, Toronto (1983) pp 215–228.

    Google Scholar 

  24. Koziel MG, Adams TL, Hazlet MA, Damm D, Miller J, Dahlbeck D, Jayne S, Staskawicz BJ: A cauliflower mosaic virus promoter directs expression of kanamycin resistance in morphogenic transformed plant cells. J Molec Appl Genet 2: 549–562 (1984).

    Google Scholar 

  25. Barker RF, Idler KB, Thompson DV, Kemp JD: Nucleotide sequence of the T-DNA region from the Agrobacterium tumefaciens octopine Ti plasmid 15955. Plant Molec Biol 2: 335–350 (1983).

    Google Scholar 

  26. Firoozabady E, DeBoer DL: Isolation, culture, and cell division in cotyledon protoplasts of cotton (Gossypium hirsutum and G. barbadense). Plant Cell Rep 5: 127–131 (1986).

    Google Scholar 

  27. Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).

    Google Scholar 

  28. Stewart JMcD, Hsu CL: In ovulo embryo culture and seedling development of cotton (Gossypium hirsutum L.). Planta 137: 113–117 (1977).

    Google Scholar 

  29. Christou P, Platt SG, Ackerman MC: Opine synthesis in wild-type plant tissue. Plant Physiol 82: 218–221 (1986).

    Google Scholar 

  30. Guesdon J, Ternynck T, Avrameas S: The use of avidin-biotin interaction in immunoenzymatic techniques. J Histochem Cytochem 27: 1131–1139 (1979).

    Google Scholar 

  31. Murai N, Sutton DW, Murray MG, Slightom JL, Merlo DJ, Reichert NA, Sengupta-Gopalan C, Stock CA, Barker RF, Kemp JD, Hall TC: Phaseolin gene from bean is expressed after transfer to sunflower via Ti plasmid vectors. Science 222: 476–482 (1983).

    Google Scholar 

  32. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72: 248–254 (1976).

    Google Scholar 

  33. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).

    Google Scholar 

  34. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354 (1979).

    Google Scholar 

  35. Gilead Z, Jeng Y-H, Wold WSM, Sugawara K, Rho HM, Harter ML, Green M: Immunological identification of two adenovirus 2-induced early proteins possibly involved in cell transformation. Nature 264: 263–266 (1976).

    Google Scholar 

  36. Blake MS, Johnston KH, Russell-Jones GJ, Gotschlich CE: A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on western blots. Anal Biochem 136: 175–179 (1984).

    Google Scholar 

  37. Murray MG, Kennard WC: Altered chromatin conformation of the higher plant gene phaseolin. Biochemistry 23: 4225–4232 (1984).

    Google Scholar 

  38. Southern EM: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517 (1975).

    Google Scholar 

  39. Balazs E, Bouzoubaa S, Guilley H, Jonard G, Paszkowski J, Richards K: Chimeric vector construction for higher-plant transformation. Gene 40: 343–348 (1985).

    Google Scholar 

  40. Beck E, Ludwig G, Auerswald EA, Reiss B, Schaller H: Nucleotide sequence and exact localization of the neomycin-phosphotransferase gene from transposon Tn5. Gene 19: 327–336 (1982).

    Google Scholar 

  41. Franck A, Guilley H, Jonard G, Richards K, Hirth L: Nucleotide sequence of cauliflower mosaic virus DNA. Cell 21: 285–294 (1980).

    Google Scholar 

  42. Oka A, Sugisaki H, Takanami M: Nucleotide sequence of the kanamycin resistance transposon Tn903. J Mol Biol 147: 217–226 (1981).

    Google Scholar 

  43. Mazodier P, Cossar P, Giraud E, Gasser F: Completion of the nucleotide sequence of the central region of Tn5 confirms the presence of three resistance genes. Nucl Acids Res 13: 195–205 (1985).

    Google Scholar 

  44. Sutcliff JG: Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harbor Symp Quant Biol 43: 77–90 (1978).

    Google Scholar 

  45. Sukhapinda K, Spivey R, Simpson RB, Shahin EA: Transgenic tomato (Lycopersicon esculentum L.) transformed with a binary vector in Agrobacterium rhizogenes: Non-chimeric origin of callus clone and low copy numbers of integrated vector T-DNA. Mol Gen Genet 206: 491–497 (1987).

    Google Scholar 

  46. Firoozabady E, Galbraith DW: Presence of a plant cell wall is not required for transformation of Nicotiana by Agrobacterium tumefaciens. Plant Cell Tissue Organ Cult 3: 175–188 (1984).

    Google Scholar 

  47. Firoozabady E, Galbraith DW: The effects of cell wall and cell cycle on crown gall tumorigenesis. HortScience 18: 441 (1983).

    Google Scholar 

  48. Otten LABM, Schilperoort RA: A rapid micro scale method for the detection of lysopine and nopaline dehydrogenase activities. Biochim Biophys Acta 527: 497–500 (1978).

    Google Scholar 

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Authors and Affiliations

  1. Agrigenetics Advanced Science Company, 5649 E. Buckeye Road, 53716, Madison, WI, USA

    Ebrahim Firoozabady, David L. DeBoer, Donald J. Merlo, Edward L. Halk, Lorraine N. Amerson, Kay E. Rashka & Elizabeth E. Murray

Authors
  1. Ebrahim Firoozabady
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  2. David L. DeBoer
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  3. Donald J. Merlo
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  4. Edward L. Halk
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  5. Lorraine N. Amerson
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  6. Kay E. Rashka
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  7. Elizabeth E. Murray
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Firoozabady, E., DeBoer, D.L., Merlo, D.J. et al. Transformation of cotton (Gossypium hirsutum L.) by Agrobacterium tumefaciens and regeneration of transgenic plants. Plant Mol Biol 10, 105–116 (1987). https://doi.org/10.1007/BF00016148

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  • DOI: https://doi.org/10.1007/BF00016148

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