Summary
The autonomous mobile element Activator from Zea mays was introduced into Arabidopsis thaliana via Agrobacterium-mediated gene transfer. The use of a chimaeric construct, where the Ac element is located in the leader of the neomycin phosphotransferase (NPT II) gene, enabled the excision of Ac to be monitored by assaying for the reconstitution of NPT II gene activity. Using this approach, the transpositional activity of AC was initially studied in primary transformants. About 50% of the regenerating Ac transformants showed evidence for excision of the element. Reintegration of Ac was confirmed by Southern blot analysis. Transposition events are transmitted to the F1 generation with a minimal frequency of 0.3%. In a few exceptional cases they are detected in a high proportion of the F1 generation. Seedlings from the F2 and F3 generations were assayed for the rate of germinal excisions by scoring for kanamycin resistance. The minimal frequency of germinal excision events amounts to 0.2%–0.5% and hence allows the use of the Ac element for gene tagging purposes in A. thaliana.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Amasino RM (1986) Acceleration of nucleic acid hybridization rate by polyethylene glycol. Anal Biochem 152:304–307
Baker B, Schell J, Lörz H, Fedoroff N (1986) Transposition of the maize controlling element “Activator” in tobacco. Proc Natl Acad Sci USA 83:4844–4848
Baker B, Coupland G, Fedoroff N, Starlinger P, Schell J (1987) Phenotypic assay for excision of the maize controlling element Ac in tobacco. EMBO J 6:1547–1554
Brink RA, Williams E (1973) Mutable R-Navajo alleles of cyclic origin in maize. Genetics 73:273–296
Estelle MA, Somerville CR (1986) The mutants of Arabidopsis. Trends Genet 2:89–93
Fedoroff NV, Furtek DB, Nelson OE (1984) Cloning of the bronze locus in maize by a simple and generalizable procedure using the transposable controlling element Activator (Ac). Proc Natl Acad Sci USA 81:3825–3829
Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Feldmann KA, Marks MD, Christianson ML, Quatrano RS (1989) A dwarf mutant of Arabidopsis generated by T-DNA insertion mutagenesis. Science 243:1351–1354
Greenblatt IM (1968) The mechanism of Modulator transposition in maize. Genetics 58:585–597
Hain R, Stabel P, Czernilofsky AP, Steinbiß HH, Herrera-Estrella L, Schell J (1985) Uptake, integration, expression and genetic transmission of a selectable chimaeric gene by plant protoplasts. Mol Gen Genet 199:161–168
Jones JDG, Carland FM, Maliga P, Dooner HK (1989) Visual detection of transposition of the maize element Activator (Ac) in tobacco seedlings. Science 244:204–207
Knapp S, Coupland G, Uhrig H, Starlinger P, Salamini F (1988) Transposition of the maize transposable element Ac in Solanum tuberosum. Mol Gen Genet 213:285–290
Lassner MW, Palys JM, Yoder JI (1989) Genetic transactivation of Dissociation elements in transgenic tomato plants. Mol Gen Genet 218:25–32
Larkin PJ, Scowcroft WR (1981) Somaclonal variation — a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:197–214
Leutwiler LS, Hough-Evans BR, Meyerowitz EM (1984) The DNA of Arabidopsis thaliana. Mol Gen Genet 194:15–23
Martin C, Carpenter R, Sommer H, Saedler H, Coen ES (1985) Molecular analysis of instability in flower pigmentation of Antirrhinum majus, following isolation of the pallida locus by transposon tagging. EMBO J 4:1625–1630
Meyerowitz EM, Pruitt RE (1985) Arabidopsis thaliana and plant molecular genetics. Science 229:1214–1218
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325
O'Reilly C, Shepherd NS, Pereira A, Schwarz-Sommer Z, Bertram I, Robertson DS, Peterson PA, Saedler H (1985) Molecular cloning of the a1 locus of Zea mays using the transposable elements En and Mu1. EMBO J 4:877–882
Pang PP, Meyerowitz EM (1987) Arabidopsis thaliana: A model system for plant molecular biology. Biotechnology 5:1177–1181
Paz-Ares J, Wienand U, Peterson PA, Saedler H (1986) Molecular cloning of the c locus of Zea mays: a locus regulating the anthocyanin pathway. EMBO J 5:829–833
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
Schmidt R, Willmitzer L (1988) High efficiency Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana leaf and cotyledon explants. Plant Cell Rep 7:583–586
Schreier PH, Seftor EA, Schell J, Bohnert HJ (1985) The use of nuclear encoded sequences to direct the light regulated synthesis and transport of a foreign protein into plant chloroplasts. EMBO J 4:25–32
Theres N, Scheele T, Starlinger P (1987) Cloning of the Bz2 locus of Zea mays using the transposable element Ds as a gene tag. Mol Gen Genet 209:193–197
Van Sluys MA, Tempé J, Fedoroff N (1987) Studies on the introduction and mobility of the maize Activator element in Arabidopsis thaliana and Daucus carota. EMBO J 6:3881–3889
Yoder JI, Palys J, Alpert K, Lassner M (1988) Ac transposition in transgenic tomato plants. Mol Gen Genet 213:291–296
Author information
Authors and Affiliations
Additional information
Communicated by H. Saedler
Rights and permissions
About this article
Cite this article
Schmidt, R., Willmitzer, L. The maize autonomous element Activator (Ac) shows a minimal germinal excision frequency of 0.2%–0.5% in transgenic Arabidopsis thaliana plants. Molec. Gen. Genet. 220, 17–24 (1989). https://doi.org/10.1007/BF00260850
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00260850