Skip to main content
SpringerLink
Log in

Somatic intrachromosomal homologous recombination events in populations of plant siblings

  • Research Article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Intrachromosomal homologous recombination in whole tobacco plants was analyzed using β-glucuronidase as non-selectable marker. We found that recombination frequencies were additive for transgenes in allelic positions and could be enhanced by treatment of plants with DNA-damaging agents. We compared the patterns of distribution of recombination events of different transgenic lines of tobacco and Arabidopsis with the respective Poisson distributions. Some lines showed Poisson-like distributions, indicating that recombination at the transgene locus was occurring in a random fashion in the plant population. In other cases, however, the distributions deviated significantly from Poisson distributions indicating that for specific transgene loci and/or configurations recombination events are not randomly distributed in the population. This was due to overrepresentation of plants with especially many as well as especially few recombination events. Analysis of one tobacco line indicated furthermore that the distribution of recombination events could be influenced by treating the seedlings with external factors. Our results suggest that different plant individuals, or parts of them, might exhibit different transient ‘states’ of recombination competence. A possible model relating ‘recombination silencing’ and transcription silencing to heterochromatization of the transgene locus is discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Allshire RC, Javerzat J-P, Redhead NJ, Cranston G: Position effect variegation at fission yeast centromers. Cell 76: 157–169 (1994).

    Article  PubMed  Google Scholar 

  2. Assaad FA, Signer ER: Somatic and germinal recombination of a direct repeat in Arabidopsis. Genetics 132: 553–566 (1992).

    PubMed  Google Scholar 

  3. Assaad FA, Tucker KL, Signer ER: Epigenetic repeat-induced gene silencing (RIGS) in Arabidopsis. Plant Mol Biol 22: 1067–1085 (1993).

    PubMed  Google Scholar 

  4. Bennett MD, Smith JB: Nuclear DNA amounts in angiosperms. Phil Trans R Soc Lond B274: 227–274 (1976).

    Google Scholar 

  5. Bevan M: Binary Agrobacterium vectors for plant transformation. Nucl Acids Res 12: 8711–8721 (1984).

    PubMed  Google Scholar 

  6. Brooks Low K: Genetic recombination: a brief overview. In: Brooks Low K (ed) The Recombination of Genetic Material, pp. 1–18. Academic Press, San Diego (1988).

    Google Scholar 

  7. Charlesworth B, Sniegowski P, Stephan W: The evolutionary dynamics of repetitive DNA in eukaryotes. Nature 371: 215–220 (1994).

    Article  PubMed  Google Scholar 

  8. Cochran WG: Some methods for strengthening the common chi-square tests. Biometrics 10: 417–451 (1954).

    Google Scholar 

  9. Das OP, Levi-Minzi S, Koury M, Benner M, Messing J: A somatic gene rearrangement contribution to genetic diversity in maize. Proc Natl Acad Sci USA 87: 7809–7813 (1990).

    PubMed  Google Scholar 

  10. deCarvalho F, Geysen G, Kushnir S, VanMontagu M, Inze D, Castresana C: Suppression of β-1,3-glucanase transgene expression in homozygous plants. EMBO J 11: 2595–2602 (1992).

    PubMed  Google Scholar 

  11. Dorer DR, Henikoff S: Expansions of transgene repeats cause heterochromatin formation and gene silencing in Drosophila. Cell 77: 993–1002 (1994).

    Article  PubMed  Google Scholar 

  12. Flavell RB: Repeated sequences and genome change. In: Hohn B, Dennis E (eds) Plant Gene Research, vol. 2. Genetic Flux in Plants, pp. 139–156. Springer-Verlag, Wien/New York (1985).

    Google Scholar 

  13. Gal S, Pisan B, Hohn T, Grimsley N, Hohn B: Genomic homologous recombination in planta. EMBO J 10: 1571–1578 (1991).

    PubMed  Google Scholar 

  14. Gangloff S, Lieber MR, Rothstein R: Transcription, topoisomerases and recombination. Experientia 50: 261–269 (1994).

    PubMed  Google Scholar 

  15. Grossenbacher-Grunder AM: Spontaneous mitotic recombination in Schizoaccharomyces pombe. Curr Genet 10: 95–101 (1985).

    Google Scholar 

  16. Henikoff S: Position-effect variegation after 60 years. Trends Genet 6: 422–426 (1990).

    Article  PubMed  Google Scholar 

  17. Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT: A simple and general method for transferring genes into plants. Science 227: 1229–1231 (1985).

    Google Scholar 

  18. Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).

    Google Scholar 

  19. Jorgenson R: Altered gene expression in plants due to trans interactions between homologous genes. Trends Biotechnol 8: 340–344 (1990).

    Article  PubMed  Google Scholar 

  20. Lebel EG, Masson J, Bogucki A, Paszkowski J: Stress-induced intrachromosomal recombination in plant somatic cells. Proc Natl Acad Sci USA 90: 422–426 (1993).

    PubMed  Google Scholar 

  21. Lichtenstein C, Paszkowski J, Hohn B: Intrachromosomal recombination between genomic repeats. In: Paszkowski J (ed) Homologous Recombination and Gene Silencing in Plants, pp. 95–122. Kluwer Academic Publishers, Dordrecht (1994).

    Google Scholar 

  22. Mattanovich D, Rüker F, deCamara Machado A, Laimer M, Regner F, Steinkellner H, Himmler G, Katinger H: Efficient transformation of Agrobacterium ssp. by electroporation. Nucl Acids Res 17: 6747 (1989).

    PubMed  Google Scholar 

  23. Matzke M, Matzke AJM, Mittelsten Scheid O: Inactivation of repeated genes: DNA-DNA interaction. In: Paszkowski J (ed) Homologous Recombination and Gene Silencing in Plants, pp. 271–307. Kluwer Academic Publishers, Dordrecht (1994).

    Google Scholar 

  24. Matzke AJM, Neuhuber F, Park Y-D, Ambros PF, Matzke MA: Homology-dependent gene silencing in transgenic plants: epistatic silencing loci contain multiple copies of methylated transgenes. Mol Gen Genet 244: 219–229 (1994).

    Google Scholar 

  25. Meins F, Kunz C: Silencing of chitinase expression in transgenic plants: an autoregulatory model. In: Paszkowski J (ed) Homologous Recombination and Gene Silencing in Plants, pp. 335–348. Kluwer Academic Publishers, Dordrecht (1994).

    Google Scholar 

  26. Meyer P, Heidmann I, Niedenhof I: Differences in DNA-methylation are associated with a paramutation phenomenon in transgenic petunia. Plant J 4: 89–100 (1993).

    Article  PubMed  Google Scholar 

  27. Mittelsten Scheid O, Afsar K, Paszkowski J: Gene inactivation in Arabidopsis thaliana is not accompanied by an accumulation of repeat-induced point mutations. Mol Gen Genet 244: 325–330 (1994).

    Article  PubMed  Google Scholar 

  28. Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).

    Google Scholar 

  29. Neuhaus J-M, Ahl-Goy P, Hinz U, Flores S, Meins F: High-level expression of a tobacco chitinase gene in Nicotiana sylvestris. Susceptibility of transgenic plants to Cercospora nicotianae infection. Plant Mol Biol 16: 141–151 (1991).

    Google Scholar 

  30. Neuhuber F, Park Y-D, Matzke AJM, Matzke MA: Susceptibility of transgene loci to homology-dependent gene silencing. Mol Gen Genet 244: 230–241 (1994).

    Article  PubMed  Google Scholar 

  31. Peterhans A, Schlüpmann H, Basse C, Paszkowski J: Intrachromosomal recombination in plants. EMBO J 9: 3437–3445 (1990).

    PubMed  Google Scholar 

  32. Puchta H, Hohn B: A transient assay in plant cells reveals a positive correlation between extrachromosomal recombination rates and length of homologous overlap. Nucl Acids Res 19: 2693–2700 (1991).

    PubMed  Google Scholar 

  33. Puchta H, Kocher S, Hohn B: Extrachromosomal homologous DNA recombination in plant cells is fast and is not affected by CpG methylation. Mol Cell Biol 12: 3372–3379 (1992).

    PubMed  Google Scholar 

  34. Puchta H, Dujon B, Hohn B: Homologous recombination in plant cells is enhanced by in vivo induction of double strand breaks into DNA by a site-specific endonuclease. Nucl Acids Res 21: 5034–5040 (1993).

    PubMed  Google Scholar 

  35. Puchta H, Swoboda P, Hohn B: Homologous recombination in plants. Experientia 50: 277–284 (1994).

    Google Scholar 

  36. Puchta H, Swoboda P, Hohn B: Induction of intrachromosomal homologous recombination in whole plants. Plant J 7, 203–210 (1995).

    Article  Google Scholar 

  37. Ramanathan B, Smerdon MJ: Enhanced DNA repair synthesis in hyperacetylated nucleosomes. J Biol Chem 264: 11026–11034 (1989).

    PubMed  Google Scholar 

  38. Säll T, Nilsson N-O, Bengtsson BO: When everyone's map is different. Curr Biol 3: 631–633 (1993).

    Article  PubMed  Google Scholar 

  39. Schmid B: Phenotypic variation in plants. Evol Trends Plants 6: 45–60 (1992).

    Google Scholar 

  40. Simpson GG, Roe A, Lewontin RC: Quantitative Zoology. Harcort, Brace and World, New York (1960).

    Google Scholar 

  41. Smith GR: Hotspots of homologous recombination. Experientia 50: 234–241 (1994).

    PubMed  Google Scholar 

  42. Sokal RR, Rohlf FJ: Biometry. Freeman, San Francisco (1973).

    Google Scholar 

  43. Swoboda P, Hohn B, Gal S: Somatic homologous recombination in planta: the recombination frequency is dependent on the allelic state of the recombining sequences and may be influenced by genomic position effects. Mol Gen Genet 237: 33–40 (1993).

    Article  PubMed  Google Scholar 

  44. Swoboda P, Gal S, Hohn B, Puchta H: Intrachromosomal homologous recombination in whole plants. EMBO J 13: 484–489 (1994).

    PubMed  Google Scholar 

  45. Tovar J, Lichtenstein C: Somatic and meiotic chromosomal recombination between inverted duplications in transgenic tobacco plants. Plant Cell 4: 319–332 (1992).

    Article  PubMed  Google Scholar 

  46. Walbot R: On life strategies of plants and animals. Trends Genet 1: 165–169 (1985).

    Article  Google Scholar 

  47. Walter V: Grundlagen der Pflanzenkunde, Das Verhaltenerbgleicher Pflanzen; die Zufallskurve. Ulmer Verlag, Stuttgart, pp. 401–407 (1962).

    Google Scholar 

Download references

Author information

Author notes

  1. Holger Puchta

    Present address: Institut für Pflanzengenetik und Kulturpflanzenforschung, Correnstrasse 3, D-06466, Gatersleben, Germany

Authors and Affiliations

  1. Friedrich Miescher-Institut, P.O. Box 2543, CH-4002, Basel, Switzerland

    Holger Puchta, Peter Swoboda, Susannah Gal & Barbara Hohn

  2. Department of Microbiology, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056, Basel, Switzerland

    Michel Blot

  3. Department of Genetics, University of Washington, 98195, Seattle, WA, USA

    Peter Swoboda

  4. Department of Biological Sciences, The State University of New York at Binghamton, 13902-6000, Binghamton, NY, USA

    Susannah Gal

Authors
  1. Holger Puchta
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter Swoboda
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Susannah Gal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Michel Blot
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Barbara Hohn
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Puchta, H., Swoboda, P., Gal, S. et al. Somatic intrachromosomal homologous recombination events in populations of plant siblings. Plant Mol Biol 28, 281–292 (1995). https://doi.org/10.1007/BF00020247

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00020247

Key words

Search

Navigation