The 5S DNA sequences in Triticum tauschii are organised in large clusters containing units that are primarily either 420 ("short") or 490 base pairs (bp) in length ("long"). The main cluster of short units was shown to be located on chromosome 1D in hexaploid wheat and is designated 5SDna-D1, while the cluster of long units was shown to be on chromosome 5D and is designated 5SDna-D2. The chromosomal locations in hexaploid wheat most likely correspond to those in T. tauschii and this could be shown directly for the 5SDna-D2 locus by using a T. tauschii 5D substitution in 'Chinese Spring' wheat. The sequence alignment of units derived from 5SDna-D1 and 5SDna-D2 revealed three apparent deletions in the noncoding spacer region, which were fixed in units from 5SDna-D1, and one deletion, which was fixed in units from 5SDna-D2. A minor size class, 400 bp long and closely related to the units from 5SDna-D1, was found in 2 of 415 accessions surveyed. A continuous range of quantitative changes in the number of 5S DNA units at the two loci was evident with up to a 10-fold relative abundance level of units being found in some accessions. Triticum tauschii var. typica was particularly noteworthy in that many accessions showed more units at 5SDna-D2 relative to 5SDna-D1. Partial thermal dissociation experiments with radioactive probes, synthesized from either the short or long 5S DNA units, hybridized to genomic DNA showed that the population of units at the respective loci were relatively homogeneous and clearly distinct from each other.(ABSTRACT TRUNCATED AT 250 WORDS)