Abstract
Transient expression of chloramphenicol acetyl transferase gene has been detected in cultured barley (Hordeum vulgare L. cv. Heartland) cells and freshly isolated immature zygotic embryos (cv. Ellice) following the introduction of the gene by microprojectile bombardment. The DNA expression vector used to introduce the CAT gene, pCaMVI1CN, is a pUC8 derivative and consisted of a CaMV35S promoter, a fragment of alcohol dehydrogenase intron1, a CAT coding region and NOS polyadenylation region. The inclusion of the Adh1 intron1 was essential for the expression of CAT activity in cultured cells as well as immature zygotic embryos. Expression of CAT activity, which was dependent upon the DNA concentration used, could be detected as early as 20 h after bombardment. The results also suggested that the recipient cells have to be in an active state of cell division in order for the introduced gene to be expressed since mature zygotic as well as somatic embryos failed to reveal any gene expression. The effect of other parameters which influence the expression of the introduced gene as well as the potential of this novel technology for cereal transformation are also discussed.
Similar content being viewed by others
Abbreviations
- Adh:
-
Alcohol dehydrogenase-1
- CaMV35S:
-
Cauliflower mosaic virus promoter
- CAT:
-
Chloramphenicol acetyltransferase
- CAP:
-
Chloramphenicol
- AcCAP:
-
Acetylated chloramphenicol derivatives
- Dicamba:
-
3,6-dichloro-2-methoxybenzoic acid
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- 2,4,5-T.:
-
2,4,5-trichlorophenoxyacetic acid
References
Callis J., Fromm M, Walbot V (1987) Genes and Dev 1: 1183–2000.
Draper J, Scott R, Armitage P, Walden R (1988) Plant Genetic Transformation and Gene Expression. A Laboratory Manual. Blackwell Scientific Publications.
Gasser CS, Fraley RT (1989) Science 244: 1293–1299.
Gamborg OL, Miller RA, Ojima K (1968) Exp Cell Res 50: 151–158.
Klein TM, Fromm M, Weissinger A, Tomes D, Schaaf S, Sietten M, Sanford JC (1988) Proc Natl Acad Sci USA 85: 4305–4309.
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory. Cold Spring Harbor, NY, USA.
Oard JH, Paige D, Dvorak J (1989) Plant Cell Reports 8: 156–160.
Rhodes CA, Pierce DA, Mettler IJ, Mascarenhas D, Detmer JJ (1988) Science 240: 204–206.
Sanford JC, Klein TM, Wolf ED, Allen N (1987) Particle Sci Technol. 5: 27–37.
Shimamoto K, Terada R, Izawa T, Fujimoto H (1989) Nature 338: 274–276.
Toriyama K, Arimoto Y, Uchimiya H, Hinata K (1988) Biotechnology 6: 1072–1074.
Uchimiya H, Fushimi T, Hashimoto H, Harada H, Syono K, Sugawara Y (1986) Molec. Gen. Genet 204: 204–207.
Wang CY, Klein TM, Fromm M, Cao J, Sanford JC, Wu R (1988) Plant Molecular Biology 11: 433–439.
Author information
Authors and Affiliations
Additional information
Communicated by F. Constabel
NRCC No. 30706
Rights and permissions
About this article
Cite this article
Kartha, K.K., Chibbar, R.N., Georges, F. et al. Transient expression of chloramphenicol acetyltransferase (CAT) gene in barley cell cultures and immature embryos through microprojectile bombardment. Plant Cell Reports 8, 429–432 (1989). https://doi.org/10.1007/BF00269041
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00269041