Abstract
We have applied a simple method for evaluation of gfp gene expression in plants using a CCD camera and computerized processing of images. Transgenic tobacco plants were obtained by Agrobacterium tumefaciens-mediated transfer of plasmid T-DNA bearing a m-gfp5-ER sequence governed by the 35S promoter together with the nptII selectable marker gene. Presence of the gfp gene in plants was confirmed by a polymerase chain reaction method. Mean brightness values measured using image analysis software showed differences between transgenic and control plants and suggest the possibility of rapid selection of transgenic individuals among regenerants and their progenies.
This is a preview of subscription content, log in via an institution to check access.
Abbreviations
- GFP:
-
green fluorescent protein
- MB:
-
mean brightness
- MS:
-
Murashige and Skoog (1962) culture medium
- PCR:
-
polymerase chain reaction
References
Edwards, K., Johnstone, C., Thompson, C.: A simple and rapid method for the preparation of plant genomic DNA for PCR analysis.-Nucl. Acids Res. 19: 1349, 1991.
Halfhill, M.D., Richards, H.A., Mabon, S.A., Stewart, C.N., Jr.: Expression of GFP and Bt transgenes in Brassica napus and hybridization with Brassica rapa.-Theor. appl. Genet. 130: 659–667, 2001.
Harper, B.K., Mabon, S.A., Leffel, S.M., Halfhill, M.D., Richards, H.A., Moyer, K.A., Stewart, C.N., Jr.: Green fluorescent protein as a marker for expression of a second gene in transgenic plants.-Nature Biotechnol. 17: 1125–1129, 1999.
Horsch, R.B., Fry, J.E., Hoffmann, N.L., Eicholtz, D., Rogers, S.G., Fraley, R.T.: A simple and general method for transferring genes into plants.-Science 227: 1229–1231, 1985.
Hu, W., Cheng, C.-L.: Expression of Aequorea green fluorescent protein in plant cells.-FEBS Lett. 369: 331–334, 1995.
Maliga, P., Sz.-Breznovits, A., Márton, L.: Streptomycin-resistant plants from callus culture of haploid tobacco.-Nature 244: 29–30, 1973.
Miki, B., McHugh, S.: Selectable marker genes in transgenic plants: applications, alternatives and biosafety.-J. Biotechnol. 107: 193–232, 2004.
Millwood, R.J., Halfhill, M.D., Harkins, D., Russotti, R., Stewart, C.N., Jr.: Instrumentation and methodology for quantitative GFP fluorescence in intact transgenic plants.-Biotechniques 24: 638–643, 2003.
Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures.-Physiol. Plant. 15: 473–497, 1962.
Prasher, D.C., Eckenrode, V.K., Ward, W.W., Prendergast, F.G., Cormier, M.J.: Primary structure of the Aequorea victoria green-fluorescent protein.-Gene 111: 229–233, 1992.
Soukupová, J., Albrechtová, J.: Image analysis — tool for quantification of histochemical detections of phenolic compounds, lignin and peroxidases in neeedles of Norway spruce.-Biol. Plant. 46: 595–601, 2003.
Stewart, C.N., Jr.: The utility of green fluorescent protein in transgenic plants.-Plant Cell Rep. 20: 376–382, 2001.
Voinnet, O., Baulcombe, D.C.: Systemic signalling in gene silencing.-Nature 389: 553, 1997.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hraška, M., Rakouský, S. & Kocábek, T. Use of a simple semiquantitative method for appraisal of green fluorescent protein gene expression in transgenic tobacco plants. Biol Plant 49, 313–316 (2005). https://doi.org/10.1007/s10535-005-3316-z
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10535-005-3316-z