Abstract
Antibiotic resistance genes can act as either cell autonomous or non-cell autonomous genetic markers with which to monitor the excision of plant transposons. To convert spectinomycin resistance from a noncell autonomous resistance to cell autonomous resistance, a transit peptide for chloroplast localization from a petunia ribulose bisphosphate carboxylase (rbcS) gene was fused in-frame to the aadA gene, which confers spectinomycin and streptomycin resistance. Constructs were generated in which the expression of this chimeric gene was prevented by the presence, in the 5′ untranslated leader, of the maize transposons Activator (Ac) or Dissociation (Ds). When progeny of tobacco or tomato plants transformed with these constructs were germinated on spectinomycin-containing medium, germinally revertant and somatically variegated individuals could be distinguished.
Similar content being viewed by others
References
Chua N-H, Schmidt GW (1978) Post-translational transport into intact chloroplasts of a precursor to the small subunit of ribulose-1,5-bisphosphate carboxylase. Proc Natl Acad Sci USA 75:6110–6114
Coen ES, Carpenter R, Martin C (1986) Transposable elements generate novel spatial patterns of gene expression in Antirrhinum majus. Cell 47:285–296
Coen ES, Robbins TP, Almeida J, Hudson A, Carpenter R (1989) Consequences and mechanisms of transposition in Antirrhinum majus. In: Berg DE, Howe MM (eds) Mobile DNA ASM Press, Washington DC
Dean C, van den Elzen P, Tamaki S, Dunsmuir P, Bedbrook J (1985) Differential expression of the eight genes of the petunia ribulose bisphosphate carboxylase small subunit multi-gene family. EMBO J 4:3055–3061
Dean C, van den Elzen P, Tamaki S, Black M, Dunsmuir P, Bedbrook J (1987) Molecular characterization of the rbcS multigene family of Petunia (Mitchell). Mol Gen Genet 206:465–474
Dean C, Sjodin C, Page T, Jones JDG, Lister C (1992) Behaviour of the maize transposable element Ac in Arabidopsis thaliana L. Plant 2:69–81
Dooner HK, Belachew A (1989) Transposition pattern of the maize element Ac from the bz-m2 (Ac) allele. Genetics 122:447–457
Fedoroff NV (1989) Maize transposable elements. In: Berg DE, Howe MM, Mobile DNA. (eds) ASM Press, Washington DC, pp 375–411
Figurski DH, Helinski DR (1979) Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 76:1648–1652
Fillatti JJ, Kiser J, Rose R, Comai L (1987) Efficient transfer of a glyphosate tolerance gene into tomato using a binary Agrobacterium tumefaciens vector. Bio/technology 5:726–730
Greenblatt IM (1984) A chromosome replication pattern deduced from pericarp phenotypes resulting from movements of the transposable element, Modulator, in maize. Genetics 108:471–485
Highfield PE, Ellis RJ (1978) Synthesis and transport of the small subunit of ribulose bisphosphate carboxylase. Nature 271:420–424
Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA (1983) A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303:179–180
Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method of transferring genes into plants. Science 227:1229–1231
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
Jones JDG, Harper L, Carland F, Ralston E, Dooner H (1991) Reversion and altered variegation of an SPT::Ac allele in tobacco. Maydica 36:329–335
Jones JDG, Shlumukov L, Carland F, English J, Scofield S, Bishop G, Harrison K (1992) Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants. Transgene Res 1:285–297
Jones JDG, Jones DA, Bishop G, Harrison K, Carroll B, Scofield S (1993) Use of the maize transposons Activator and Dissociation to show that chimeric genes for resistance to phosphinotricin and spectinomycin act non cell autonomously in tobacco and tomato seedlings. Transgene Res 2:63–78
Keller J, Lim E, James DW Jr, Dooner HK (1992) Germinal and somatic activity of the maize element Activator (Ac) in Arabidopsis. Genetics 131:449–459
Keller J, Jones JDG, Harper E, Lim E, Carland F, Ralston E, Dooner HK (1993) Effects of gene dosage and sequence modification on the frequency and timing of transposition of the maize element Activator in tobacco. Plant Mol Biol 21:159–170
Kunkel T (1985) Rapid and efficient site-directed mutagenesis without phenotypic selection. Proc Natl Acad Sci USA 82:488–492
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
McClintock B (1948) Mutable loci in maize. Carnegie Inst Washington Yearb 47:155–169
McClintock B (1978) Development of the maize endosperm as revealed by clones. In: Subteleny S, Sussex I. (eds) The clonal basis of development. Academic Press, New York, pp 217–237
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assay with tobacco tissue cultures. Physiol Plant 15:473–497
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
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Spring Harbor Laboratory, Cold Spring Harbor, New York
Scofield S, Harrison KA, Nurrish SJ, Jones JDG (1992) Promoter fusions to the Ac transposase gene confer distinct patterns of Ds somatic and germinal excision in tobacco. Plant Cell 4:573–582
Scofield SR, English JJ, Jones JDG (1993) High level expression of the transposase gene from the maize transposon Activator (Ac) inhibits the excision of Dissociation (Ds) in tobacco cotyledons. Cell 75 507–517
Smeekens S, Bauerle C, Hageman J, Keegstra K, Weisbeek P (1986) The role of the transit peptide in the routing of precursors towards different chloroplast compartments. Cell 46:365–375
Spena A, Aalen RB, Schulze SC (1989) Cell autonomous behaviour of the rolC gene of Agrobacterium rhizogenes during leaf development: a visual assay for transposon excision in transgenic plants. Plant Cell 1:1157–1164
Swinburne J, Balcells L, Scofield S, Jones JDG, Coupland G (1992) Elevated levels of Ac transposase mRNA are associated with high frequencies of Ds excision in Arabidopsis. Plant Cell 4:583–592
van den Broeck G, Timko MP, Kausch AP, Cashmore AR, Van Montagu M, Herrera-Estrella L (1985) Targeting of a foreign protein to chloroplasts by fusion to the transit peptide from the small subunit of ribulose 1,5-bisphosphate carboxylase. Nature 313:358–363
Wessler SR (1988) Phenotypic diversity mediated by the maize transposable elements Ac and Spm. Science 242:399–405 1993
Yang C-H, Carroll B, Scofield S, Jones JDG, Michelmore R (1993) Transactivation of Ds elements in plants of lettuce (Lactuca sativa). Mol Gen Genet 241:389–398
Author information
Authors and Affiliations
Additional information
Communicated by H. Saedler
Rights and permissions
About this article
Cite this article
Scofield, S.R., Jones, D.A., Harrison, K. et al. Chloroplast targeting of spectinomycin adenyltransferase provides a cell-autonomous marker for monitoring transposon excision in tomato and tobacco. Molec. Gen. Genet. 244, 189–196 (1994). https://doi.org/10.1007/BF00283522
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00283522