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Chromosoma

, Volume 94, Issue 1, pp 11–18 | Cite as

Detection of a 17 kb unique sequence (T-DNA) in plant chromosomes by in situ hybridization

  • P. F. Ambros
  • M. A. Matzke
  • A. J. M. Matzke
Article

Abstract

An approach is described for the detection of a unique sequence, the T-DNA region of the Agrobacterium rhizogenes root-inducing (Ri) plasmid, in plant chromosomes by in situ hybridization. This sequence was introduced into the Crepis capillaris genome (2n=6) by infecting Crepis stem segments with A. rhizogenes. Roots growing from the infection site contain T-DNA and synthesize mannopine, which can be used as a convenient biochemical marker for T-DNA transformation. Southern analysis of DNA isolated from one transformed Crepis root line verified the presence of a single copy of T-DNA (approximate size 17 kb) per diploid Crepis genome. To localize T-DNA, both DNA and RNA probes, labelled with either tritium or biotin, were hybridized to Crepis chromosomes prepared from transformed root tips by a novel spreading method. Biotinylated probes were visualized using reflection-contrast microscopy. In the hybridization experiments described, T-DNA was detected in one homologue of chromosome 3, where it could be assigned to a paracentromeric position in the neighbourhood of the nucleolar organizing region. These results demonstrate that it is possible to localize unique sequences in plant chromosomes by in situ hybridization.

Keywords

Developmental Biology Biotin Tritium Single Copy Biochemical Marker 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • P. F. Ambros
    • 1
  • M. A. Matzke
    • 1
  • A. J. M. Matzke
    • 1
  1. 1.Institute of Molecular BiologyAustrian Academy of SciencesSalzburgAustria

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