Abstract
Transposition of the Anthirrinum majus Tam3 element and the Zea mays Ac element has been monitored in petunia and tobacco plants. Plant vectors were constructed with the transposable elements cloned into the leader sequence of a marker gene. Agrobacterium tumefaciens-mediated leaf disc transformation was used to introduce the transposable element constructs into plant cells. In transgenic plants, excision of the transposable element restores gene expression and results in a clearly distinguishable phenotype. Based on restored expression of the hygromycin phosphotransferase II (HPTII) gene, we established that Tam3 excises in 30% of the transformed petunia plants and in 60% of the transformed tobacco plants. Ac excises from the HPTII gene with comparable frequencies (30%) in both plant species. When the β-glucuronidase (GUS) gene was used to detect transposition of Tam3, a significantly lower excision frequency (13%) was found in both plant species. It could be shown that deletion of parts of the transposable elements Tam3 and Ac, removing either one of the terminal inverted repeats (TIR) or part of the presumptive transposase coding region, abolished the excision from the marker genes. This demonstrates that excision of the transposable element Tam3 in heterologous plant species, as documented for the autonomous element Ac, also depends on both properties. Southern blot hybridization shows the expected excision pattern and the reintegration of Tam3 and Ac elements into the genome of tobacco plants.
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Baker B, Schell J, Lörz H, Fedoroff N: Transposition of the maize controlling element ‘Activator’ in tobacco. Proc Natl Acad Sci USA 83: 4844–4848 (1986).
Baker B, Coupland G, Fedoroff N, Starlinger P, Schell J: Phenotypic assay for excision of the maize controlling element Ac in tobacco. EMBO J 6: 1547–1554 (1987).
Baulcombe DC, Saunders GR, Bevan MW, Mayo MA, Harrison BD: Expression of biologically active viral satellite RNA from the nuclear genome of transformed plants. Nature 321: 446–449 (1986).
Behrens U, Fedoroff N, Laird A, Muller-Neumann M, Starlinger P, Yoder J: Cloning of Zea mays controlling element Ac from the wx-m7 allele. Mol Gen Genet 194: 346–347 (1984).
Carpenter R, Martin C, Coen ES: Comparison of genetic behaviour of the transposable element Tam3 at two und liked pigment loci in Antirrhinum majus Mol Gen Genet 207: 82–89 (1987).
Cone KC, Burr FC, Burr B: Molecular analysis of the maize anthocyanin regulatory locus C1. Proc Natl Acad Sci USA 83: 9631–9635 (1986).
Day A, Schirmer-Rahire M, Kuchka MR, Mayfield SP, Rochaix J-D: A transposon with an unusual arrangement of long terminal repeats in the green alga Chlamydomonas reinhardtii. EMBO J 7: 1917–1927 (1988).
Dellaporta SL, Woods J, Hicks JB: A plant DNA minipreparation version II. Plant Mol Biol Rep 1(4): 19–21 (1983).
DeVlaming P: Genotype determination of Petunia Mitchell with respect to some flower color genes. Plant Mol Biol News II: 106–108 (1981).
Ditta G, Stanfield S, Corbin D, Helinski DR: Broad host range DNA cloning system for Gram-negative bacteria: Construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 12: 7347–7351 (1980).
Fedoroff NV, Furtek DB, Nelson OE Jr: 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 (1984).
Gielen J, DeBeuckeleer M, Seurinck J, Deboeck F, DeGreve H, Lemmers M, Montagu Mvan, Schell J: The complete nucleotide sequence of the TL-DNA of the Agrobacterium tumefaciens plasmid pTiAch5. EMBO J 3: 835–846 (1984).
Goddard JM, Caput D, Williams SR, Martin DW: Cloning of human purine-nucleoside phosphorylase cDNA sequences by complementation in Escherichia coli. Proc Natl Acad Sci USA 80: 4281–4285 (1983).
Grandbastien M-A, Spielmann A, Caboche M: Tnt-1 a mobile retroviral like transposable element of tobacco isolated by plant cell genetics. Nature 337: 376–380 (1989).
Herrmann A, Schulz W, Hahlbrock K: Two alleles of the single-copy chalcone synthase gene in parsley differ by a transposon-like element. Mol Gen Genet 212: 93–98 (1988).
Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA: A binary vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303: 179–180 (1983).
Hudson A, Carpenter R, Coen ES: De novo activation of the transposable element Tam2 of Antirrhinum majus. Mol Gen Genet 207: 54–59 (1987).
Inzé D, Follin A, Lijsebettens Mvan, Simoens C, Genetello C, Montagu Mvan, Schell J: Genetic analysis of the individual T-DNA genes of Agrobacterium tumefaciens; further evidence that two genes are involved in indole-3-acetic acid synthesis. Mol Gen Genet 194: 265–274 (1984).
Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).
Jones JDG, Dunsmuir P, Bedbrook J: High level expression of introduced chimaeric genes in regenerated transformed plants. EMBO J 4: 2411–2418 (1985).
Knapp S, Coupland G, Uhrig H, Starlinger P, Salamini F: Transposition of the maize transposable element Ac in Solanum tuberosum. Mol Gen Genet 213: 285–290 (1988).
Maliga P, Breznovitz A, Marton L: Streptomycin resistant plants from callus culture of resistant tobacco. Nature 244: 29–30 (1973).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Martin C, Carpenter R, Sommer H, Saedler H, Coen ES: Molecular analysis of instability in flower pigmentation of Antirrhinum majus, following isolation of the pallida locus by transposon tagging. EMBO J 4: 1625–1630 (1985).
Motto M, Maddaloni M, Ponziani G, Brembilla M, Marotta R, Di Fonzo N, Soave C, Thompson R, Salamini F: Molecular cloning of the o2-m5 allele of Zea mays using transposon marking. Mol Gen Genet 212: 488–494 (1988).
Nevers P, Shepard N, Saedler H: Plant transposable elements. Adv Bot Res 12: 103–203 (1986).
O'Reilly C, Shepherd NS, Pereira A. Schwarz-Sommer Z, Bertram I, Robertson DS, Peterson PA, Saedler H: Molecular cloning of the al locus of Zea mays using the transposable elements En and Mul. EMBO J 4: 877–882 (1985).
Paz-Ares J, Wienand U, Peterson PA, Saedler H: Molecular cloning of the c locus regulating the anthocyanin pathway. EMBO J 5: 829–833 (1986).
Sanger F, Nicklen S, Coulson R: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1980).
Shirsat AH: A transposon-like structure in the 5′ flanking sequence of a legumin gene from Pisum sativum. Mol Gen Genet 212: 129–133 (1988).
Sommer H, Carpenter R, Harrison BJ, Saedler H: The transposable element Tam3 of Antirrhinum majus generates a novel type of sequence alterations upon excision. Mol Gen Genet 199: 225–231 (1985).
Sutton WD, Gerlach WL, Schwartz D, Peacock WJ: Molecular analysis of Ds Controlling element mutations at the Adhl locus of maize. Science 233: 1265–1269 (1984).
VanSluys MA, Tempé J, Fedoroff N: Studies on the introduction and mobility of the maize Activator element in Arabidopsis thaliana and Daucus carota. EMBO J 6: 3881–3889 (1987).
Vodkin LO, Rhodes PR, Goldberg RB: A Lectin gene insertion has the structural features of a transposable element. Cell 34: 1923–1031 (1983).
Yoder JI, Palys J, Alpert K, Lassner M: Ac transposition in transgenic tomato plants. Mol Gen Genet 213: 291–296 (1988).
Martin C, Prescott A, Lister C, MacKay A: Activity of the transposon Tam3 in antirrhinum and tobacco: possible role of DNA methylation EMBO J 8: 997–1004 (1989).
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Haring, M.A., Gao, J., Volbeda, T. et al. A comparative study of Tam3 and Ac transposition in transgenic tobacco and petunia plants. Plant Mol Biol 13, 189–201 (1989). https://doi.org/10.1007/BF00016137
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DOI: https://doi.org/10.1007/BF00016137