Abstract
Transgenic wheat plants without the selectable marker gene were obtained either in the presence or in the absence of selective pressure during the transformation protocol. When using hygromycin as selective agent in a co-transformation experiment involving a mixture of plasmids pGL2, containing the hpt gene, and pAI1Gus, containing the uidA gene, 3 out of 19 transgenic wheat plants had the uidA gene alone as shown by Southern blots. The gene was transmitted to the progeny following Mendelian rules. Segregation and loss of the selectable marker gene was also found in three out of six events from other experiments where high-molecular-weight glutenin genes were expressed or over-expressed. On the other hand, in 7 experiments where no selective pressure was applied and that involved 1016 bombarded explants, 23 transgenic wheat plants were obtained. The uidA gene was stably integrated as suggested by its transmission to the progeny.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alvarez, M.L., Guelman, S., Halford, N.G., Lustig, S., Reggiardo, M.I., Ryabushkina, N., Shewry, P., Stein, J. and Vallejos, R.H. 2000. Silencing of HMW glutenins in transgenic wheat expressing extra HMW subunits. Theor. Appl. Genet. 100: 319–327.
Birch, R.G. 1997. Plant transformation: Problems and strategies for practical application. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48: 297–326.
Christensen, A.H., Sharrock, R.A. and Quail, P.H. 1992. Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation. Plant Mol. Biol. 18: 675–689.
Christou, P. 1996. Transformation technology. Trends Plant Sci. 1: 423–431.
Daley, M., Knauf, V.C., Summerfelt, K.R. and Turner, J.C. 1998. Co-transformation with one Agrobacterium tumefaciens strain containing two binary plasmid as a method for producing marker-free transgenic plants. Plant Cell Rep. 17: 489–496.
Ebinuma, H., Sugita, K., Matsunaga, E. and Yamakado, M. 1997. Selection of marker-free transgenic plants using the isopentenyl transferase gene. Proc. Natl. Acad. Sci. USA 94: 2117–2121.
Edwards, K., Johnstone, C. and Thompson, C. 1991. A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucl. Acids Res. 19: 1349.
Goldsbrough, A. 1992. Marker gene removal: a practical necessity? Trends Biotechnol. 10: 417.
Goldsbrough, A.P., Lastrella, C.N. and Yoder, J.I. 1993. Transposition mediated re-positioning and subsequent elimination of marker genes from transgenic tomato. Bio/technology 11: 1286–1292.
Hansen, G. and Wright, M.S. 1999. Recent advances in the transformation of plants. Trends Plant Sci. 4: 226–231.
Hohn, B., Levy, A.A. and Puchta, H. 2001. Elimination of selection markers from transgenic plants. Curr. Opin. Biotechnol. 12: 139–143.
Maniatis, T., Fritsch, E.F. and Sambrook, J. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Plainview, NY.
Odell, J., Caimi, P., Sauer, B. and Russell, S. 1990. Site-directed recombination in the genome of transgenic tobacco. Mol. Gen. Genet. 223: 369–378.
Ortiz, J.P.A., Reggiardo, M.I., Ravizzini, R.A., Altabe, S.G., Cervigni, G.D.L., Spitteler, M.A., Morata, M.M., Elías, F.E. and Vallejos, R.H. 1996. Hygromycin resistance as an efficient selectable marker for wheat stable transformation. Plant Cell Rep. 15: 877–881.
Ortiz, J.P.A., Ravizzini, R.A., Morata, M.M. and Vallejos, R.H. 1997. A rapid system for studying foreign gene expression in wheat (Triticum aestivum L.). J. Appl. Genet. 38: 123–130.
Puchta, H. 2000. Removal selectable marker genes: taking the shortcut. Trends Plant. Sci. 5: 273–274.
Reggiardo, M.I., Arana, J.L., Orsaria, L.M., Permingeat, H.R., Spitteler, M.A. and Vallejos, R.H. 1991. Transient transformation of maize tissues by microparticle bombardment. Plant Sci. 75: 237–243.
Saghai-Maroof, M.A., Soliman, K.M., Jorgensen, R.A. and Allard, R.W. 1984. Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proc. Natl. Acad. Sci. USA 81: 8014–8018.
Shimamoto, K., Terada, R., Izawa, T. and Fujimoto, H. 1989. Fertile transgenic rice plants regenerated from transformed protoplasts. Nature 338: 274–276.
Smalla, K., Bporin, S., Heuer, H., Gebhard, F., van Elsas, J.D. and Nielsen, K. 2000. Horizontal transfer of antibiotic resistance genes from transgenic plants to bacteria: are there new data to fuel the debate? In: C. Fairbaim, G. Scoles and A. McHughen (Eds.) Proceedings of the 6th International Symposium on ‘The Biosafety of Genetically Modified Organisms’ (Saskatoon, Canada), University Extension Press, Saskatoon, Canada, pp. 146–154.
Author information
Authors and Affiliations
Additional information
these authors contributed equally to the work
Rights and permissions
About this article
Cite this article
Permingeat, H.R., Alvarez, M.L., Cervigni, G.D. et al. Stable wheat transformation obtained without selectable markers. Plant Mol Biol 52, 415–419 (2003). https://doi.org/10.1023/A:1023969501440
Issue Date:
DOI: https://doi.org/10.1023/A:1023969501440