Advertisement

Molecular and General Genetics MGG

, Volume 199, Issue 2, pp 183–188 | Cite as

Direct gene transfer to cells of a graminaceous monocot

  • Ingo Potrykus
  • Michael W. Saul
  • Jirina Petruska
  • Jerzy Paszkowski
  • Raymond D. Shillito
Article

Summary

Definitive evidence is presented for the first time for stable gene transfer to cultured cells in a plant of the family Gramineae, Lolium multiflorum (Italian Ryegrass), using DNA transformation of protoplasts from a non-morphogenic cell culture. A construction consisting of expression signals from gene VI of Cauliflower Mosaic virus joined to the aminoglycoside (neomycin) phosphotransferase gene (APH(3′)II) from transposon Tn5 conferred resistance to the antibiotic G-418 to cell colonies arising from transformed protoplasts. By demonstrating a tight correlation between the resistant phenotype, the physical presence of the foreign gene and the presence of the active gene product we have shown that these colonies are true transformants and that a gene which is expressed well in dicotyle-denous plants is also expressed in cells of graminaceous monocots.

Keywords

Gene Transfer Mosaic Virus Aminoglycoside Neomycin Foreign Gene 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bennett MB, Smith JB (1976) Nuclear DNA amounts in Angiosperms. Philos Trans R Soc Lond [Biol] 274:227–274Google Scholar
  2. Bevan MW, Flavell RB, Chilton M-D (1983) A chimaeric antibiotic resistance gene as a selectable marker for plant cell transformation. Nature 304:184–187Google Scholar
  3. De Block M, Herrera-Estrella L, Van Montagu M, Schell J, Zambryski P (1984) Expression of foreign genes in regenerated plants and their progeny. EMBO J 3:1681–1689Google Scholar
  4. De Cleene M, De Ley J (1976) The host range of crown gall. Bot Rev 42:389–466Google Scholar
  5. Fraley RT, Rogers SG, Horsch RB, Sanders PR, Flick JS, Adams SP, Bittner ML, Brand LA, Fink CL, Fry JS, Galluppi GR, Goldberg SB, Hoffmann NL, Woo SC (1983) Expression of bacterial genes in plant cells. Proc Natl Acad Sci USA 80:4803–4807Google Scholar
  6. Hain R, Stabel P, Czernilofsky AP, Steinbiss HH, Herrera-Estrella L, Schell J (1985) Uptake, integration, expression and genetic transmission of a selectable chimeric gene by plant protoplasts. Mol Gen Genet 199:161–168Google Scholar
  7. Hernalsteens J-P, Thia-Toong L, Schell J, Van Montagu M (1984) An Agrobacterium-transformed cell culture from the monocot Asparagus officinalis. EMBO J 3:3039–3041Google Scholar
  8. Herrera-Estrella L, De Block M, Messens E, hernalsteens J-P, Van Montagu M, Shell J (1983) Chimeric genes as dominant selectable markers in plant cells. EMBO J 2:987–995Google Scholar
  9. Hooykaas-Van Slogteren GMS, Hooykaas PJJ, Schilperoort RA (1984) Expression of Ti plasmid genes in monocotyledonous plants infected with Agrobacterium tumefaciens. Nature 311:763–764Google Scholar
  10. Horsch RB, Fraley RT, Rogers SG, Sanders PR, Loyd A, Hoffmann N (1984) Inheritance of functional foreign genes in plants. Science 223:496–498Google Scholar
  11. Jones MGK, Dale PJ (1982) Reproducible regeneration of callus from suspension culture protoplasts of the grass Lolium multiflorum. Z Pflanzenphysiol 105:267–274Google Scholar
  12. Krens FA, Molendijk L, Wullems GJ, Schilperoort RA (1982) In vitro transformation of plant protoplasts with Ti-plasmid DNA. Nature 296:72–74Google Scholar
  13. Lörz H, Baker B, Schell J (1985) Gene transfer to cereal cells mediated by protoplast transformation. Mol Gen Genet 199:178–182Google Scholar
  14. Messing J, Vieira J (1982) A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene 19:269–276Google Scholar
  15. Paszkowski J, Shillito RD, Saul M, Mandak V, Hohn T, Hohn B, Potrykus I (1984) Direct gene transfer to plants. EMBO J 3:2717–2722Google Scholar
  16. Paszkowski J, Saul MW (1985) Direct gene transfer. Methods Enzymol (in press)Google Scholar
  17. Potrykus I, Harms CT, Lörz H (1979) Callus formation from cell culture protoplasts of corn. Theor Appl Genet 54:209–214Google Scholar
  18. Potrykus I (1980) The old problem of protoplast culture: cereals. In: Ferenczy L, Farkas GL (eds) Advances in protoplast research. Hungarian Academiy of Science Press, pp 243–254Google Scholar
  19. Potrykus I, Paszkowski J, Saul MW, Petruska J, Shillito RD (1985) Molecular and general genetics of a hybrid foreign gene introduced to tobacco by direct gene transfer. Mol Gen Genet 199:169–177Google Scholar
  20. Reiss B, Sprengel R, Will H, Schaller H (1984) A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30:211–218Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Ingo Potrykus
    • 1
  • Michael W. Saul
    • 1
  • Jirina Petruska
    • 1
  • Jerzy Paszkowski
    • 1
  • Raymond D. Shillito
    • 1
  1. 1.Friedrich Miescher InstitutBaselSwitzerland

Personalised recommendations