Plant Molecular Biology

, Volume 32, Issue 6, pp 1197–1203 | Cite as

Details of T-DNA structural organization from a transgenic Petunia population exhibiting co-suppression

  • Paul D. Cluster
  • Michael O'Dell
  • Michael Metzlaff
  • Richard B. Flavell
Short Communication


Analysis of Agrobacterium-transferred DNA (T-DNA) revealed strong correlations between transgene structures and floral pigmentation patterns from chalcone synthase (chs) co-suppression among 47 Petunia transformants. Presented here are the full details of T-DNA structural organization in that population. Sixteen transformants (34%) carried one T-DNA copy while 31 (66%) carried 106 complete and partial T-DNA elements in 54 linkage groups. Thirty linkage groups contained multiple T-DNA copies; 15 of these contained only contiguously repeated copies, 8 contained only dispersed copies and 7 contained both. Right-border inverted repeats were three times more frequent than left-border inverted or direct repeats. Large fragments of binary-vector sequences were linked to the T-DNA in seven plants.

Key words

Agrobacterium-mediated transformation binary-verector transfer direct repeats inverted repeats repeat-induced gene silencing transgene inheritance 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Chen DF, Dale PJ, Heslop-Harrison JS, Snape JW, Harwood W, Bean S, Mullineaux PM: Stability of transgenes andpresence of N6 methyladenine DNA in transformed wheat cells. Plant J 5: 429–436 (1994).Google Scholar
  2. 2.
    Deroles SC, Gardner RC: Analysis of the T-DNA structure in a large number of transgenic petunias generated by Agrobacterium-mediated transformation. Plant Mol Biol 11 365–377 (1988).Google Scholar
  3. 3.
    Flavell RB: Inactivation of gene expression in plants as a consequence of specific sequence duplication. Proc Natl Acad Sci USA 91: 3490–3496 (1994).Google Scholar
  4. 4.
    Jones JDG, Gilbert DE, Grady KL, Jorgensen RA: T-DNA structure and gene expression in petunia plants transformed by Agrobacterium tumefaciens C58 derivatives. Mol Gen Genet 207: 478–485 (1987).Google Scholar
  5. 5.
    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
  6. 6.
    Jorgensen R, Snyder C, Jones JDG: T-DNA is organized predominantly in inverted repeat structures in plants transformed with Agrobacterium tumefaciens C58 derivatives. Mol Gen Genet 207: 471–477 (1987).Google Scholar
  7. 7.
    Jorgensen RA, Cluster PD, English J, Que Q, Napoli CA: 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).Google Scholar
  8. 8.
    Maizonnier D: Cytology. In: Sink KC (ed) Petunia, pp. 21–33. Springer-Verlag, New York (1984).Google Scholar
  9. 9.
    Marano MR, Carrillo N: Chromoplast formation during tomato fruit ripening: no evidence for plastid DNA methylation. Plant Mol Biol 16: 11–19 (1991).Google Scholar
  10. 10.
    Martineau B, Voelker TA, Sanders RA: On defining T-DNA. Plant Cell 6: 1032–1033 (1994).Google Scholar
  11. 11.
    Matzke MA, Matzke AJM: How and why do plants inactivate homologous transgenes. Plant Physiol 107: 679–685 (1995).Google Scholar
  12. 12.
    McClelland M, Nelson M, Raschke E: Effect of site-specific modification on restriction endonucleases and DNA modification methyltransferases. Nucl Acids Res 22: 3640–3659 (1994).Google Scholar
  13. 13.
    Meyer P, Niedenhof I, ten Lohuis M: Evidence for cytosine methylation of non-symmetrical sequences in transgenic Petunia hybrida. EMBO J 13: 2084–2088 (1994).Google Scholar
  14. 14.
    Muller E, Brown PTH, Hartke S, Lorz H: DNA variation in tissue-culture-derived rice plants. Theor Appl Genet 80: 673–679 (1990).Google Scholar
  15. 15.
    Napoli C, Lemieux C, Jorgensen R: Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. Plant Cell 2: 279–289 (1990).Google Scholar
  16. 16.
    Ngernprasirtsiri J, Kobayashi H, Akazawa T: Trancriptional regulation and DNA methylation of nuclear genes for photosynthesis in nongreen plant cells. Proc Natl Acad Sci USA 86: 7919–7923 (1989).Google Scholar
  17. 17.
    Pintor-Toro JA: Adenine methylation in zein genes. Biochem Biophysical Res Comm 147: 1082–1087 (1987).Google Scholar
  18. 18.
    Zambryski P: Chronicles from the Agrobacterium-plant cell DNA transfer story. Annu Rev Plant Physiol Plant Mol Biol 43: 465–490 (1992).Google Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Paul D. Cluster
    • 1
  • Michael O'Dell
    • 1
  • Michael Metzlaff
    • 1
  • Richard B. Flavell
    • 1
  1. 1.John Innes CentreNorwichUK

Personalised recommendations