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.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Allshire RC, Javerzat J-P, Redhead NJ, Cranston G: Position effect variegation at fission yeast centromers. Cell 76: 157–169 (1994).
Assaad FA, Signer ER: Somatic and germinal recombination of a direct repeat in Arabidopsis. Genetics 132: 553–566 (1992).
Assaad FA, Tucker KL, Signer ER: Epigenetic repeat-induced gene silencing (RIGS) in Arabidopsis. Plant Mol Biol 22: 1067–1085 (1993).
Bennett MD, Smith JB: Nuclear DNA amounts in angiosperms. Phil Trans R Soc Lond B274: 227–274 (1976).
Bevan M: Binary Agrobacterium vectors for plant transformation. Nucl Acids Res 12: 8711–8721 (1984).
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).
Charlesworth B, Sniegowski P, Stephan W: The evolutionary dynamics of repetitive DNA in eukaryotes. Nature 371: 215–220 (1994).
Cochran WG: Some methods for strengthening the common chi-square tests. Biometrics 10: 417–451 (1954).
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).
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).
Dorer DR, Henikoff S: Expansions of transgene repeats cause heterochromatin formation and gene silencing in Drosophila. Cell 77: 993–1002 (1994).
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).
Gal S, Pisan B, Hohn T, Grimsley N, Hohn B: Genomic homologous recombination in planta. EMBO J 10: 1571–1578 (1991).
Gangloff S, Lieber MR, Rothstein R: Transcription, topoisomerases and recombination. Experientia 50: 261–269 (1994).
Grossenbacher-Grunder AM: Spontaneous mitotic recombination in Schizoaccharomyces pombe. Curr Genet 10: 95–101 (1985).
Henikoff S: Position-effect variegation after 60 years. Trends Genet 6: 422–426 (1990).
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).
Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).
Jorgenson R: Altered gene expression in plants due to trans interactions between homologous genes. Trends Biotechnol 8: 340–344 (1990).
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).
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).
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).
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).
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).
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).
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).
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).
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).
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).
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).
Peterhans A, Schlüpmann H, Basse C, Paszkowski J: Intrachromosomal recombination in plants. EMBO J 9: 3437–3445 (1990).
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).
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).
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).
Puchta H, Swoboda P, Hohn B: Homologous recombination in plants. Experientia 50: 277–284 (1994).
Puchta H, Swoboda P, Hohn B: Induction of intrachromosomal homologous recombination in whole plants. Plant J 7, 203–210 (1995).
Ramanathan B, Smerdon MJ: Enhanced DNA repair synthesis in hyperacetylated nucleosomes. J Biol Chem 264: 11026–11034 (1989).
Säll T, Nilsson N-O, Bengtsson BO: When everyone's map is different. Curr Biol 3: 631–633 (1993).
Schmid B: Phenotypic variation in plants. Evol Trends Plants 6: 45–60 (1992).
Simpson GG, Roe A, Lewontin RC: Quantitative Zoology. Harcort, Brace and World, New York (1960).
Smith GR: Hotspots of homologous recombination. Experientia 50: 234–241 (1994).
Sokal RR, Rohlf FJ: Biometry. Freeman, San Francisco (1973).
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).
Swoboda P, Gal S, Hohn B, Puchta H: Intrachromosomal homologous recombination in whole plants. EMBO J 13: 484–489 (1994).
Tovar J, Lichtenstein C: Somatic and meiotic chromosomal recombination between inverted duplications in transgenic tobacco plants. Plant Cell 4: 319–332 (1992).
Walbot R: On life strategies of plants and animals. Trends Genet 1: 165–169 (1985).
Walter V: Grundlagen der Pflanzenkunde, Das Verhaltenerbgleicher Pflanzen; die Zufallskurve. Ulmer Verlag, Stuttgart, pp. 401–407 (1962).
Author information
Authors and Affiliations
Rights 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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00020247