Abstract
The cauliflower mosaic virus promoter is commonly used to drive transcription of chimeric genes in transgenic plants, including the cereals. To determine the tissue and cell types of cereal plants that the promoter functions in, transgenic rice plants containing a CaMV 35S promoter/GUS chimeric gene were analyzed for GUS activity. Insertion of a 35S/GUS chimeric gene at low copy number into chromosomal DNA of plants regenerated from electroporated protoplasts was confirmed by gel blot hybridization analysis of uncut and endonuclease-digested DNA. Quantitative measurement showed that GUS activity was some tenfold higher in rice leaves than in tobacco leaves [8] whereas activities obtained for rice roots were similar to those reported for tobacco roots. Histochemical localization of GUS activity confirmed that the CaMV 35S promoter functions in cells of the leaf epidermis, mesophyll and vascular bundle. It is also active in the cortex and vascular cylinder of the root, but only marginally active in the root epidermis. The generally similar distribution and levels of GUS activity obtained in differentiated tissue of stably transformed rice plants indicates the value of the CaMV 35S promoter as a positive control for studies in gene activity in transgenic monocots and dicots.
Similar content being viewed by others
References
Benfey PN, Chua N-H: Regulated genes in transgenic plants. Science 244: 174–181 (1989).
Benfey PN, Ren L, Chua N-H: The CaMV 35S enhancer contains at least two domains which confer different developmental and tissue-specific expression patterns. EMBO J 8: 2195–2202 (1989).
Bennet MD, Smith JB: Nuclear DNA amounts in angiosperms. Philos Trans R Soc London, Ser B 274: 227–274 (1976).
Bradford MM: A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72: 248–254 (1976).
Fromm M, Taylor LP, Walbot V: Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci USA 82: 5824–5828 (1985).
Fromm ME, Taylor LP, Walbot V: Stable transformation of maize after gene transfer by electroporation. Nature 319: 791–793 (1986).
Hauptmann RM, Ozias-Akins P, Vasil V, Tabaeizadeh Z, Rogers SG, Horsch RB, Vasil IK, Fraley RT: Transient expression of electroporated DNA in monocotyledonous and dicotyledonous species. Plant Cell Rep 6: 265–270 (1987).
Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).
Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: β-glucuronidase as a versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).
Labarca C, Paigen K: A simple, rapid, and sensitive DNA assay procedure. Anal Biochem 102: 344–352 (1980).
Lee B, Murdoch K, Topping J, Martin K, Jones MGK: Transient gene expression in aleurone protoplasts isolated from developing caryopses of barley and wheat. Plant Mol Biol 13: 21–29 (1989).
Lyznik LA, Ryan RD, Ritchie SW, Hodges TK: Stable co-transformation of maize protoplasts with the gusA and neo genes. Plant Mol Biol 13: 151–161 (1989).
Marcotte WR, Bayley CC, Quatrano RS: Regulation of a wheat promoter by abscisic acid in rice protoplasts. Nature 335: 454–457 (1988).
Marcotte WR, Russell SH, Quatrano RS: Abscisic acid-responsive sequences from the Em gene of wheat. Plant Cell 1: 969–976 (1989).
Ou-Lee T-M, Turgeon R, Wu R: Expression of a foreign gene linked to either a plant-virus or a Drosophila promoter, after electroporation of protoplasts of rice, wheat, and sorghum. Proc Natl Acad Sci USA 83: 6815–6819 (1986).
Reed KC: Evaluation of Southern transfers. Mol Biol Rep 4: 3–4 (1988).
Reed KC, Mann DA: Rapid transfer of DNA from agarose to nylon membranes. Nucleic Acids Res 13: 7207–7221 (1985).
Rhodes CA, Pierce DA, Mettler IJ, Mascarenhas D, Detmer JJ: Genetically transformed maize plants from protoplasts. Science 240: 204–207 (1988).
Shimamoto K, Terada RT, Izawa T, Fujimoto H: Fertile transgenic rice plants regenerated from transformed protoplasts. Nature 338: 274–276 (1989).
Taylor B, Powell A: Isolation of plant DNA and RNA. Focus 4: 4–6 (1982).
Terada R, Shimamoto K: Expression of CaMV35S-GUS gene in transgenic rice plants. Mol Gen Genet 220: 389–392 (1990).
Toriyama K, Arimoto Y, Uchimiya H, Hinata K: Transgenic rice plants after direct gene transfer into protoplasts. Bio/Technology 6: 1072–1074 (1988).
Yang H, Zhang HM, Davey MR, Mulligan BJ, Cocking EC: Production of kanamycin resistant rice tissue following DNA uptake into protoplasts. Plant Cell Rep 7: 421–425 (1988).
Zhang EM, Yang H, Rech EL, Golds TJ, Davis AS, Milligan BJ, Cocking EC, Davey MR: Transgenic rice plants produced by electroporation-mediated plasmid uptake into protoplasts. Plant Cell Rep 7: 379–384 (1988).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Battraw, M.J., Hall, T.C. Histochemical analysis of CaMV 35S promoter-β-glucuronidase gene expression in transgenic rice plants. Plant Mol Biol 15, 527–538 (1990). https://doi.org/10.1007/BF00017828
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00017828