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Transgenic tomato lines containing Ds elements at defined genomic positions as tools for targeted transposon tagging

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We have introduced a genetically marked Dissociation transposable element (Ds HPT) into tomato (Lycopersicon esculentum) by Agrobacterium tumefaciens-mediated transformation. Probes for the flanking regions of the T-DNA and transposed Ds HPT elements were obtained with the inverse polymerase chain reaction (IPCR) technique and used in RFLP linkage analyses. The RFLP map location of 11 T-DNAs carrying Ds HPT was determined. The T-DNAs are distributed on 7 of the 12 tomato chromosomes. To explore the feasibility of gene tagging strategies in tomato using Ds HPT, we examined the genomic distribution of Ds HPT receptor sites relative to the location of two different, but very closely linked, T-DNA insertion sites. After crosses with plants expressing Ac transposase, the hygromycin phosphotransferase (HPT) marker on the Ds element and the excision markers β-glucuronidase (GUS) and Basta resistance (BAR) facilitated the identification of plants bearing germinally transposed Ds HPT elements. RFLP mapping of 21 transposed Ds HPT elements originating from the two different T-DNA insertions revealed distinct patterns of reintegration sites.

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  1. Baker B, Schell J, Lörz H, Fedoroff NV (1986) Transposition of the maize controlling element Activator in tobacco. Proc Natl Acad Sci USA 83:4844–4848

  2. Bancroft I, Jones JDG, Dean C (1993) Heterologous transposon tagging of the DRL1 locus in Arabidopsis. Plant Cell 5:631–638

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

  4. Brandstädter J, Roßbach C, Theres K (1994) The pattern of histone H4 expression in the tomato shoot apex changes during development. Planta 192:69–74

  5. 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

  6. De Block M, Botterman J, Vandewiele M, Dockx J, Thoen C, Gossele V, Rao Movva N, Thompson C, Van Montagu M, Leemans J (1987) Engineering herbicide resistance in plants by expression of a detoxifying enzyme. EMBO J 6:2513–2518

  7. Dellaporta SL, Greenblatt I, Kermicle JL, Hicks JB, Wessler SL (1988) Molecular cloning of the maize R-nj allele by transposon tagging with Ac. Stadler Symp. 18:263–282

  8. Dooner HK, Belachew A (1989) Transposition pattern of the maize element Ac from the bz-m2(Ac) allele. Genetics 122:447–475

  9. 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

  10. Fedoroff NV, Furtek DB, Nelson OE (1984) Cloning the bronze locus in maize by a simple and generalizable procedure using the controlling element Activator (Ac). Proc Natl Acad Sci USA 81:3829–3835

  11. Fillatti J, Kiser J, Rose R, Comai L (1987) Efficient transfer of a glyphosphate tolerance gene into tomato using a binary Agrobacterium tumefaciens vector. Biotechnology 5:726–728

  12. Greenblatt IM (1984) A chromosomal replication pattern deduced from pericarp phenotypes resulting from movements of the transposable element Modulator in maize. Genetics 108:471–485

  13. Healy J, Corr C, DeYoung J, Baker B (1993) Linked and unlinked transposition of a genetically marked Dissociation element in transgenic tomato. Genetics 134:571–584

  14. 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

  15. Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907

  16. Jones JDG, Carland F, 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

  17. Keller J, Lim E, Dooner HK (1993) Preferential transposition of Ac to linked sites in Arabidopsis. Theor Appl Genet 86:586–588

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

  19. 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

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

  21. Kunze R, Starlinger P (1989) The putative transposase of transposable element Ac from Zea mays L. interacts with subterminal sequences of Ac. EMBO J 8:3177–3185

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

  23. Lassner MW, Palys JM, Yoder JI (1989) Genetic transactivation of Dissociation elements in transgenic tomato plants. Mol Gen Genet 218:25–32

  24. 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

  25. Martin GB, de Vincente MC, Tanksley SD (1993) High-resolution linkage analysis and physical characterization of the Pto bacterial resistance locus in tomato. Mol Plant-Microbe Interact 6:26–34

  26. Masterson RV, Furtek DB, Grevelding C, Schell J (1989) A maize Ds transposable element containing a dihydrofolate reductase gene transposes in Nicotiana tabacum and Arabidopsis thaliana. Mol Gen Genet 219:461–466

  27. Meissner R (1990) Versuche zur Inhibition eines Gens von Lycopersicon esculentum mit Hille von “antisense” RNA. Diplomathesis, Universität zu Köln, Köln

  28. Müller-Neumann M, Yoder JI, Starlinger P (1984) The DNA sequence of the transposable element Ac of Zea mays L. Mol Gen Genet 198:19–24

  29. Ochman H, Gerber AS, Hard DL (1988) Genetic applications of an inverse polymerase chain reaction. Genetics 120:621–623

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

  31. Rommens CMT, Rudenko GN, Dijkwel PP, van Haaren MJJ, Ouwerkerk PBF, Blok KM, Nijkamp HJJ, Hille J (1992) Characterization of the Ac/Ds behaviour in transgenic tomato plants using plasmid rescue. Plant Mol Biol 20:61–70

  32. Rommens CMT, Munyikwa TRI, Overduin B, Nijkamp HJJ, Hille J (1993) Transposition pattern of a modified Ds element in tomato. Plant Mol Biol 21:1109–1119

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

  34. Scofield S, Harrison KA, Nurrish SJ, Jones JDG (1992) Promotor fusions to the Ac transposase gene confer distinct patterns of Ds somatic and germinal excision in tobacco. Plant Cell 4:573–582

  35. Stevens MA, Rick CM (1986) Genetics and breeding. In: Atherton JG, Rudich J (eds) The tomato crop. Chapman and Hall, New York, pp 35–109

  36. Tanksley SD, Ganal MW, Prince JP, Vicente MC, Bonierbale MW, Broun P, Fulton TM, Giovannoni JJ, Grandillo S, 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

  37. 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

  38. 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 16:81–86

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

  40. Yoder JI (1990b) A genetic analysis of mutations recovered from tomato following Agrobacterium-mediated transformation with the maize transposable elements Activator and Dissociation. Theor Appl Genet 79:657–662

  41. Yoder JI, Palys J, Alpert K, Lassner M (1988) Ac transposition in transgenic tomato plants. Mol Gen Genet 213:291–296

  42. Van Sluys MA, Tempe J, Fedoroff NV (1987) Studies on the introduction and mobility of the maize Activator in Arabidopsis thaliana and Daucus carota, EMBO J 6:3881–3889

  43. Velten J, Velten L, Hain R, Schell J (1984) Isolation of a dual promoter fragment from the Ti plasmid of Agrobacterium tumefaciens. EMBO J 3:2723–2730

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Correspondence to Klaus Theres.

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Communicated by H. Saedler

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Knapp, S., Larondelle, Y., Roßberg, M. et al. Transgenic tomato lines containing Ds elements at defined genomic positions as tools for targeted transposon tagging. Molec. Gen. Genet. 243, 666–673 (1994). https://doi.org/10.1007/BF00279576

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Key words

  • Lycopersicon esculentum
  • Ac/Ds
  • Transposon tagging
  • Inverse PCR
  • RFLP
  • linkage analysis