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


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1986-07

AUTHORS

P. F. Ambros, M. A. Matzke, A. J. M. Matzke

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. More... »

PAGES

11-18

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf00293525

DOI

http://dx.doi.org/10.1007/bf00293525

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1049582904


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