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Analysis of the chromosomal distribution of transposon-carrying T-DNAs in tomato using the inverse polymerase chain reaction

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Abstract

We are developing a system for isolating tomato genes by transposon mutagenesis. In maize and tobacco, the transposon Activator (Ac) transposes preferentially to genetically linked sites. To identify transposons linked to various target genes, we have determined the RFLP map locations of Ac- and Dissociation (Ds)-carrying T-DNAs in a number of transformants. T-DNA flanking sequences were isolated using the inverse polymerase chain reaction (IPCR) and located on the RFLP map of tomato. The authenticity of IPCR reaction products was tested by several criteria including nested primer amplification, DNA sequence analysis and PCR amplification of the corresponding insertion target sequences. We report the RFLP map locations of 37 transposon-carrying T-DNAs. We also report the map locations of nine transposed Ds elements. T-DNAs were identified on all chromosomes except chromosome 6. Our data revealed no apparent chromosomal preference for T-DNA integration events. Lines carrying transposons at known map locations have been established which should prove a useful resource for isolating tomato genes by transposon mutagenesis.

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References

  • Aarts MGM, Dirkse WG, Stiekma WJ, Pereira A (1993) Transposon tagging of a male sterility gene in Arabidopsis. Nature 363:715–717

    Google Scholar 

  • Ambros PF, Matzke AJM, Matzke MA (1986) Localization of Agrobacterium rhizogenes T-DNA in plant chromosomes by in situ hybridization. EMBO J 5:2073–2077

    Google Scholar 

  • 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

    Google Scholar 

  • Bacroft I, Bhatt AM, Sjodin S, Scofield S, Jones JDG, Dean C (1992) Development of an efficient two-element transposon tagging system in Arabidopsis thaliana. Mol Gen Genet 233:449–461

    Google Scholar 

  • Barker RF, Idler KB, Thompson DV, Kemp JD (1983) Nucleotide sequence of the T-DNA region from the Agrobacterium tumefaciens octopine Ti plasmid pTi15955. Plant Mol Biol 2:335–350

    Google Scholar 

  • Barnes DA, Thorner J (1986) Genetic manipulation of Saccharomyces cerevisiae by use of the lys-2 gene. Mol Cell Biol 6:2828–2838

    Google Scholar 

  • Belzile F, Yoder JI (1992) Pattern of somatic transposition in a high copy Ac tomato line. Plant J 2:173–179

    Google Scholar 

  • Chuck G, Robbins T, Nijjar C, Ralston E, Courtney-Gutterson N, Dooner HK (1993) Tagging and cloning of a petunia flower color gene with the maize transposable element Activator. Plant Cell 5:371–378

    Google Scholar 

  • Chyi Y-S, Jorgensen RA, Goldstein D, Tanksley SD, Loaiza-Figueroa F (1986) Locations and stability of Agrobacterium-mediated T-DNA insertions in the Lycopersicon genome. Mol Gen Genet 204:64–69

    Google Scholar 

  • Dean C, Sjodin C, Page T, Jones JDG, Lister C (1992) Behaviour of the maize transposable element Ac in Arabidopsis thaliana L. Plant J 2:69–81

    Google Scholar 

  • Dickinson MJ, Jones DA, Jones JDG (1993) Close linkage between the Cf-2/Cf-5 and Mi resistance loci in tomato. Mol Plant Microbe Interact 6:341–347

    Google Scholar 

  • Dooner HK, Belachew A (1989) Transposition pattern of the maize element Ac from the bz-m2 (Ac) allele. Genetics 122:447–457

    Google Scholar 

  • Dooner HK, Keller J, Harper E, Ralston E (1991) Variable patterns of transposition of the maize element Activator in tobacco. Plant Cell 3:473–482

    Google Scholar 

  • Earp DJ, Lowe B, Baker B (1990) Amplification of genomic sequences flanking transposable elements in host and heterologous plants: a tool for transposon tagging and genome characterization. Nucleic Acids Res 18:3271–3279

    Google Scholar 

  • English JJ, Harrison K, Jones JDG (1993) A genetic dissection of DNA sequence requirements for Dissociation State I activity in tobacco. Plant Cell 5:501–514

    Google Scholar 

  • Ganal MW, Lapitan NLV, Tanksley SD (1988) A molecular and cytogenetic survey of major repeated DNA sequences in tomato (Lycopersicon esculentum). Mol Gen Genet 213:262–268

    Google Scholar 

  • 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

    Google Scholar 

  • Hehl R, Baker B (1990) Properties of the maize transposable element Activator in transgenic tobacco plants: a versatile interspecies genetic tool. Plant Cell 2:709–721

    Google Scholar 

  • Herman L, Jacobs A, Van Montagu M (1990) Plant chromosome/marker gene fusion assay for study of normal and truncated T-DNA integration events. Mol Gen Genet 224:248–256

    Google Scholar 

  • Jacobs JP, Yoder JI (1989) Ploidy levels in transgenic tomato plants determined by chloroplast number. Plant Cell Rep 7:662–664

    Google Scholar 

  • 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

    Google Scholar 

  • Jones JDG, Carland FC, Lim E, Ralston E, Dooner HK (1990) Preferential transposition of the maize element Activator to linked chromosomal locations in tobacco. Plant Cell 2:701–707

    Google Scholar 

  • Jones JDG, Shlumukov L, Carland FJ, Scofield S, Bishop G, Harrison K (1992a) Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants. Transgene Res 1:285–297

    Google Scholar 

  • Jones JDG, Bishop G, Carroll B, Dickinson M, English J, Harrison K, Jones D, Scofield S, Thomas CM (1992b) Prospects for establishing a tomato gene tagging system using the maize transposon Activator (Ac). Proc R Soc Edinburgh 99B (3/4):107–119

    Google Scholar 

  • Jones DA, Dickinson MJ, Balint-Kurti PJ, Dixon MS, Jones JDG (1993a) Two complex resistance loci revealed in tomato by classical and RFLP mapping of the Cf-2, Cf-4, Cf-5 and Cf-9 genes for resistance to Cladosporium fulvum. Mol Plant Microbe Interact 6:348–357

    Google Scholar 

  • Jones JDG, Jones DA, Bishop G, Harrison K, Carroll B, Scofield S (1993b) Use of the maize transposons Activator and Dissociation to show that chimeric genes for resistance to phosphinothricin and spectinomycin act non cell autonomously in tobacco and tomato seedlings. Transgene Res 2:63–78

    Google Scholar 

  • 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 21:157–170

    Google Scholar 

  • Klimyuk VI Carroll BJ, Thomas CM, Jones JDG (1993) Alkali treatment for rapid preparation of plant tissue for reliable PCR analysis. Plant J 3:493–494

    Google Scholar 

  • 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

    Google Scholar 

  • Koncz C, Martini N, Mayerhofer R, Koncz-Kalman Z, Koerber H, Redei GP, Schell J (1989) High-frequency T-DNA-mediated gene tagging in plants. Proc Natl Acad Aci USA 86:8467–8471

    Google Scholar 

  • Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln SE, Newburn L (1987) MAPMAKER: An interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174–181

    Google Scholar 

  • Martin C, Prescott A, Lister C, MacKay S (1989) Activity of the transposon Tam3 in Antirrhinum and tobacco: possible role of DNA methylation EMBO J 8:997–1004

    Google Scholar 

  • Ochmann H, Gerber AS, Hartl DL (1988) Genetic applications of an inverse polymerase chain reaction. Genetics 120:621–623

    Google Scholar 

  • Osborne BI, Corr CA, Prince JP, Hehl R, Tanksley SD, McCormick S, Baker B (1991) Ac transpositions from a T-DNA can generate linked and unlinked clusters of insertions in the tomato genome. Genetics 129:833–844

    Google Scholar 

  • Ralston E, English J, Dooner HK (1989) Chromosome-breaking structure in maize involving a fractured Ac element. Proc Natl Acad Sci USA 86:9451–9455

    Google Scholar 

  • Rick CM, Yoder JI (1988) Classical and molecular genetics of tomato: highlights and perspectives. Annu Rev Genet 22:281–300

    Google Scholar 

  • Rosenthal A, Jones DSC (1990) Genomic walking and sequencing by oligo-cassette mediated polymerase chain reaction. Nucleic Acids Res 18:3095–3096

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Schmidt R, Willmitzer L (1989) The maize autonomous element Activator (Ac) shows a minimal germinal excision frequency of 0.2%–0.5% in transgenic Arabidopsis thaliana. Mol Gen Genet 220:17–24

    Google Scholar 

  • Scofield S, Harrison K, 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

    Google Scholar 

  • Sherman JD, Stack SM (1992) Two-dimensional spreads of synaptonemal complexes from solanaceous plants. V. Tomato (Lycopersicon esculentum) karyotype and idiogram. Genome 35:354–359

    Google Scholar 

  • Shyamala V, Ferro-Luzzi Ames G (1989) Genomic walking by single-specific-primer polymerase chain reaction: SSP-PCR*. Gene 84:1–8

    Google Scholar 

  • 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

    Google Scholar 

  • Swinburne J, Balcells L, Scofield S, Jones JDG, Coupland G (1992) Elevated levels of Ac transposase mRNA are associated with high frequencies of Ds escision in Arabidopsis. Plant Cell 4:583–592

    Google Scholar 

  • Tanksley SD, Ganal MW, Prince JP, de Vicente MC, Bonierbale MW, Broun P, Fulton TM, Giovanonni JJ, Grandillo GB, Martin GB, Messeguer R, Miller JC, Miller L, Paterson AH, Pineda O, Röder MS, Wing RA, Wu W, Young ND (1992) High density molecular linkage maps of the tomato and potato genomes. Genetics 132:1141–1160

    Google Scholar 

  • Triglia T, Peterson MG, Kemp DJ (1988) A procedure for in vitro amplification of DNA segments that lie outside the boundaries of known sequences. Nucleic Acids Res 16:8186

    Google Scholar 

  • Walbot V (1992) Strategies for mutagenesis and gene cloning using transposon tagging and T-DNA insertional mutagenesis. Annu Rev Plant Physiol Plant Mol Biol 43:49–82

    Google Scholar 

  • Wallroth M, Gerats AM, Rogers SG, Fraley RT, Horsch RB (1986) Chromosomal localization of foreign genes in Petunia hybrida. Mol Gen Genet 202:6–15

    Google Scholar 

  • Wang K, Stachel SE, Timmerman B, Van Montagu M, Zambryski PC (1987) Site-specific nick in the T-DNA border sequence as a result of Agrobacterium vir gene expression. Science 235:587–590

    Google Scholar 

  • Waye MMY, Verhoeyen ME, Jones PT, Winter G (1985) EcoK selection vectors for shotgun cloning into M13 and deletion mutagenesis. Nucleic Acids Res 13:8561–8571

    Google Scholar 

  • Yoder JI (1990) Rapid proliferation of the maize transposable element Activator in transgenic tomato. Plant Cell 2:723–730

    Google Scholar 

  • Zamir D, Tanksley SD (1988) Tomato genome is comprised largely of fast-evolving, low copy-number sequences. Mol Gen Genet 213:254–261

    Google Scholar 

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Authors and Affiliations

  1. The Sainsbury Laboratory, John Innes Centre, NR4 7UH, Colney Lane, Norwich, UK

    Colwyn M. Thomas, David A. Jones, James J. English, Bernard J. Carroll, Kate Harrison & Jonathan D. G. Jones

  2. Department of Biological Sciences, Purdue University, 47907, West Lafayette, Indiana, USA

    Jeffrey L. Bennetzen

  3. Department of Physiology and Environmental Science, University of Nottingham, Faculty of Agricultural and Food Sciences, Sutton Bonnington, Nr. Loughborough, Leicestershire, UK

    Alan Burbidge

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  1. Colwyn M. Thomas
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  2. David A. Jones
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  3. James J. English
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  4. Bernard J. Carroll
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  5. Jeffrey L. Bennetzen
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  6. Kate Harrison
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  7. Alan Burbidge
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  8. Gerard J. Bishop
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  9. Jonathan D. G. Jones
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Communicated by D.M. Lonsdale

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Thomas, C.M., Jones, D.A., English, J.J. et al. Analysis of the chromosomal distribution of transposon-carrying T-DNAs in tomato using the inverse polymerase chain reaction. Molec. Gen. Genet. 242, 573–585 (1994). https://doi.org/10.1007/BF00285281

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  • DOI: https://doi.org/10.1007/BF00285281

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