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We review and extend data showing concerted evolution of parental 18–5.8–26S nuclear ribosomal DNA (18–26S rDNA) gene families in three natural Nicotiana allotetraploids (N. tabacum, N. rustica and N. arentsii, each 2n = 4x = 48) and one synthetic N. tabacum line (Th37, ♀ N. sylvestris (2n = 24) × ♂N. tomentosiformis (2n = 24)). The origin of the gene families was analysed by sequence polymorphisms in the intergenic spacer (IGS) region and the number of chromosomal loci by fluorescence in situ hybridization (FISH). FISH revealed that the number and locations of 18–26S rDNA in the natural allopolyploids was the sum of those found in the diploid progenitors. However, the rDNA restriction patterns showed polymorphisms in the IGS that were not additive, suggesting that parental rDNA clusters were partially (N. tabacum, N. rustica) or completely (N. arentsii) overwritten by hybrid-specific units. Thus …