Molecular cytogenetic characterisation of the terminal heterochromatic segment of the B-chromosome of rye (Secale cereale) |
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| Authors: | Andreas Houben Ralf G Kynast Ute Heim Hanno Hermann R Neil Jones John W Forster |
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| Institution: | (1) Institut fur Pflanzengenetik und Kulturpflanzenforschung, D-06466 Gatersleben, Germany;(2) Genetics Group, Institute of Biological Sciences, University of Wales, Aberystwyth, SY 23 3DD Dyfed, Wales, UK;(3) Present address: Department of Genetics, The University of Adelaide, 5005, South Australia, Australia |
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| Abstract: | The terminal heterochromatic segments of the long arms of 20 rye B-chromosomes were isolated by means of laser microdissection
technology. Also the remaining portions of the long arms, along with the short arms of the same chromosomes were isolated.
Each sample was used for degenerate oligonucleotide primer-polymerase chain reaction (DOP-PCR) amplification reactions. The
resulting products were used as probes for chromosome in situ hybridisation experiments, and in Southern hybridisation to
digests of 0B and +B DNA. Competition hybridisation of these probes with 0B DNA allowed the detection of B-specific sequences.
The terminal heterochromatin of the rye B-chromosome contains both B-specific sequences and sequences also present on the
A-chromosomes of rye. The B-specific D1100 family is the major repeat species located in the terminal heterochromatin. Primers
designed to the cloned sequence (E1100) were used to search for related low copy sequences in 0B DNA. The sequences of the
PCR products revealed no similarities to that of the clone E1100 except for the primer sequences. The possible origin of this
sequence is discussed in the context of models for the evolution of the rye B-chromosome.
EMBL and Genbank accession numbers: Z54196 (E1199): Z54278 (B1)
Edited by: R. Appels |
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