Chalcone synthase cosuppression phenotypes in petunia flowers: comparison of sense vs. antisense constructs and single-copy vs. complex T-DNA sequences (original) (raw)

Abstract

Flower pigmentation patterns were scored in 185 sense_Chalcone synthase (Chs)_ transgenotes and 85 antisense_Chs_ transgenotes; upon first flowering, 139 (75%) of sense transgenotes were found to be phenotypically altered, as were 70 (82%) of the antisense transgenotes. The observed patterns document the range of phenotypic variations that occur, as well as confirm and extend the finding that sense_Chs_ constructs produce several types of morphologybased based flower pigmentation patterns that antisense_Chs_ constructs do not. Long-term monitoring for epigenetic variations in one population of 44 sense_Chs_ transgenotes showed that 43 (98%) were capable of producing a cosuppression phenotype. The primary determinant of sense-specific patterns of cosuppression of_Chs_ was found to be the repetitiveness and organization pattern of the transgene, not ‘position effects’ by, or ‘readthrough’ from, flanking plant DNA sequences. The degree of cosuppression observed in progeny of transgenotes carrying multiple, dispersed copies as compared to that observed with a single copy of the transgene suggests that sense cosuppression of_Chs_ is subject to a transgene dosage effect.

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References

  1. de Carvalho F, Gheysen G, Kushnir S, Van Montagu 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
  2. de Carvalho Niebel F, Frendo P, Van Montagu M, Cornelissen M: Post-transcriptional cosuppression of β-1,3-glucanase genes does not affect accumulation of nuclear mRNA. Plant Cell 7: 347–358 (1995).
    Article PubMed Google Scholar
  3. Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: version II. Plant Mol Biol Rep 1: 19–21 (1983).
    Google Scholar
  4. 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
  5. Grierson D, Fray RG, Hamilton AJ, Smith CJS, Watson CF: Does co-suppression of sense genes in transgenic plants involve antisense RNA? Trends Biotechnol 9: 122–123 (1991).
    Article Google Scholar
  6. Jones JDG, Shlumukov L, Carland F, English J, Scofield SR, Bishop GJ, Harrison K: Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants. Transgen Res 1: 285–297 (1992).
    Google Scholar
  7. Jorgensen R: Elicitation of organized pigmentation patterns by a chalcone synthase transgene. In: Amasino RM (ed) Cellular Communication in Plants. pp. 87–92. Plenum Press, New York (1993).
    Google Scholar
  8. Jorgensen R: Developmental significance of epigenetic impositions on the plant genome: a paragenetic function for chromosomes. Devel Genet 15: 523–532 (1994).
    Google Scholar
  9. Jorgensen RA: Cosuppression, flower color patterns, and metastable gene expression states. Science 268: 686–691 (1995).
    Google Scholar
  10. Jorgensen RA, Napoli CA: A responsive regulatory system is revealed by sense suppression of pigment genes in_Petunia_ flowers. In: Gustafson JP, Flavell RB (eds) Genomes, Proceedings of the 22nd Stadler Genetics Symposium, Plenum Press, New York (in press).
  11. Lindbo JA, Silva-Rosales L, Proebsting WM, Dougherty WG: Induction of a highly specific antiviral state in transgenic plants: implications for gene regulation and virus resistance. Plant Cell 5: 1749–1759 (1993).
    Article PubMed Google Scholar
  12. Matzke MA, Primig M, Tronovsky J, Matzke AJM: Reversible methylation and inactivation of marker genes in sequentially transformed tobacco plants. EMBO J 8: 643–649 (1989).
    Google Scholar
  13. Matzke MA, Matzke AJM: How and why do plants inactivate homologous transgenes. Plant Physiol 107: 679–685 (1995).
    PubMed Google Scholar
  14. 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).
    Article PubMed Google Scholar
  15. MeinsJr. F, Kunz C: Gene silencing in transgenic plants: a heuristic autoregulation model. Curr Topics Microbiol Immunol 197: 105–120 (1994).
    Google Scholar
  16. Murray MG, Thompson WF: Isolation of high molecular weight plant DNA. Nucleic Acids Res 8: 4321–4325 (1980).
    PubMed Google Scholar
  17. Napoli C, Lemieux C, Jorgensen R: Introduction of a chimeric chalcone synthase gene into petunia results in reversible cosuppression of homologous genes_in trans_. Plant Cell 2: 279–289 (1990).
    Article PubMed Google Scholar
  18. Taylor LP, Jorgensen R: Conditional male fertility in chalcone synthase-deficient petunia. J Hered 83: 11–17 (1992).
    Google Scholar
  19. van Blokland R, van der Geest N, Mol JNM, Kooter JM: Transgene-mediated suppression in_Petunia hybrida_ results from an increase in RNA turnover. Plant J 6: 861–877 (1994).
    Article Google Scholar

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Author notes

  1. Paul D. Cluster
    Present address: CNR Instituto di Ricerche sul Miglioramento Genetico delle Piante Foraggere, Via Madonna Alta 130, 06128, Perugia, Italy
  2. James English
    Present address: Sainsbury Laboratory, John Innes Centre, NR4 7UH, Norwich, UK

Authors and Affiliations

  1. Environmental Horticulture, University of California, 95616-8587, Davis, CA, USA
    Richard A. Jorgensen, Paul D. Cluster, James English, Qiudeng Que & Carolyn A. Napoli

Authors

  1. Richard A. Jorgensen
  2. Paul D. Cluster
  3. James English
  4. Qiudeng Que
  5. Carolyn A. Napoli

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Jorgensen, R.A., Cluster, P.D., English, J. et al. Chalcone synthase cosuppression phenotypes in petunia flowers: comparison of sense vs. antisense constructs and single-copy vs. complex T-DNA sequences.Plant Mol Biol 31, 957–973 (1996). https://doi.org/10.1007/BF00040715

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