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.
Similar content being viewed by others
References
Bennett MB, Smith JB (1976) Nuclear DNA amounts in Angiosperms. Philos Trans R Soc Lond [Biol] 274:227–274
Bevan MW, Flavell RB, Chilton M-D (1983) A chimaeric antibiotic resistance gene as a selectable marker for plant cell transformation. Nature 304:184–187
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 3:1681–1689
De Cleene M, De Ley J (1976) The host range of crown gall. Bot Rev 42:389–466
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 80:4803–4807
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 199:161–168
Hernalsteens J-P, Thia-Toong L, Schell J, Van Montagu M (1984) An Agrobacterium-transformed cell culture from the monocot Asparagus officinalis. EMBO J 3:3039–3041
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 2:987–995
Hooykaas-Van Slogteren GMS, Hooykaas PJJ, Schilperoort RA (1984) Expression of Ti plasmid genes in monocotyledonous plants infected with Agrobacterium tumefaciens. Nature 311:763–764
Horsch RB, Fraley RT, Rogers SG, Sanders PR, Loyd A, Hoffmann N (1984) Inheritance of functional foreign genes in plants. Science 223:496–498
Jones MGK, Dale PJ (1982) Reproducible regeneration of callus from suspension culture protoplasts of the grass Lolium multiflorum. Z Pflanzenphysiol 105:267–274
Krens FA, Molendijk L, Wullems GJ, Schilperoort RA (1982) In vitro transformation of plant protoplasts with Ti-plasmid DNA. Nature 296:72–74
Lörz H, Baker B, Schell J (1985) Gene transfer to cereal cells mediated by protoplast transformation. Mol Gen Genet 199:178–182
Messing J, Vieira J (1982) A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene 19:269–276
Paszkowski J, Shillito RD, Saul M, Mandak V, Hohn T, Hohn B, Potrykus I (1984) Direct gene transfer to plants. EMBO J 3:2717–2722
Paszkowski J, Saul MW (1985) Direct gene transfer. Methods Enzymol (in press)
Potrykus I, Harms CT, Lörz H (1979) Callus formation from cell culture protoplasts of corn. Theor Appl Genet 54:209–214
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 243–254
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 199:169–177
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 30:211–218
Author information
Authors and Affiliations
Additional information
Communicated by H. Saedler
Dedicated to professor Georg Melchers to celebrate his 50-year association with the journal
Rights and permissions
About this article
Cite this article
Potrykus, I., Saul, M.W., Petruska, J. et al. Direct gene transfer to cells of a graminaceous monocot. Molec Gen Genet 199, 183–188 (1985). https://doi.org/10.1007/BF00330257
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00330257