Hypervariation associated with a 12-nucleotide direct repeat and inferences on intergenomic homogenization of ribosomal RNA gene spacers based on the DNA sequence of a clone from the wheat Nor-D3 locus

Abstract

A ribosomal RNA gene (rDNA) unit from the Nor-D3 locus (D genome) of Triticum aestivum L. was cloned and the "nontranscribed spacer" (NTS) was sequenced. The DNA sequence was compared with previously reported Nor-B2 locus (B genome) NTS sequences to study the molecular basis of evolution of these repeated genes and to look for evidence of homogenization between B- and D-genome rDNA. The NTS has seven subrepeats with a modal repeat length of 120 nucleotides; the subrepeats are shorter than Nor-B2 subrepeats owing to loss of one element of a 12-bp duplication present in Nor-B2 subrepeats. This 12 nucleotide sequence or its permutation, whose consensus sequence is CACGTACACGGA, is found at all sites where the B- and D-genome rDNA spacers differ by insertions or deletions longer than two nucleotides. The DNA sequence information was used to identify restriction sites unique to each locus that could be used in search of conversions between the B- and D-genome rDNA loci. Despite the coexistence of rDNA of the B- and D-genomes in the same nucleus for a minimum of 8000 years, no evidence for frequent interchromosomal conversion events between chromosomes 1B or 6B and 5D was found.