Plant Molecular Biology

, Volume 5, Issue 4, pp 235–246 | Cite as

Transformation of plant protoplasts with DNA: cotransformation of non-selected calf thymus carrier DNA and meiotic segregation of transforming DNA sequences

  • R. Peerbolte
  • F. A. Krens
  • R. M. W. Mans
  • M. Floor
  • J. H. C. Hoge
  • G. J. Wullems
  • R. A. Schilperoort
Article
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Summary

With the DNA transformation procedure developed in our laboratory (13) several transformed tobacco SR1 tissues were obtained which, apart from selected and non-selected pTi sequences (T+), also had acquired non-selected calf thymus carrier DNA sequences (C+), being integrated in their nuclear genomes. From one such tissue (cNT4), with a shooty crown gall phenotype and expressing mannopine synthesis activity (Mas+), shoots were grafted and mature, flowering plants (gNT4) were obtained. After cross pollination with wild type SR1 tobacco pollen 49% of the seedlings obtained, had the maternal NT4-like crown gall phenotype and 51% showed wild type (SR1) features. The mannopine locus segregated independently from the locus determining the crown gall phenotype. When screened for integrated (‘transforming’) foreign DNA sequences 97% of the NT4-like seedlings turned out to be C+T+. Most of the SR1-like seedlings, having a wild type tobacco morphology, proved to be transformed as well: roughly a 1:1:1:1 ratio as found for C+T+:C-T+: C+T:C T SR1-like seedlings. Based on the segregation of transforming sequences during meiosis a model is presented showing the integration of these sequences in three different host chromosomes.

Keywords

cotransformation DNA-transformation N. tabacum progeny Ti plasmid 
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Copyright information

© Martinus Nijhoff Publishers 1985

Authors and Affiliations

  • R. Peerbolte
    • 1
  • F. A. Krens
    • 1
  • R. M. W. Mans
    • 1
  • M. Floor
    • 1
  • J. H. C. Hoge
    • 1
  • G. J. Wullems
    • 1
  • R. A. Schilperoort
    • 1
  1. 1.Dept. of Plant Molecular BiologyState University of LeidenLeidenThe Netherlands

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