The variation at the internal transcribed spacer (ITS) region of the ribosomal DNA has been correlated with the number of nucleolar organizer regions (NORs) in some plant species. Besides, the number of NORs might influence the rate of homogenization of the rDNA repeats. In recent studies, ITS length variants were detected in bread wheat cultivars but no reports about their presence in durum wheat were found. In the present study, we localized and identified the NORs of 51 old Portuguese durum wheat cultivars by using sequential silver staining and fluorescence in situ hybridization performed with the pTa71 rDNA probe. We also detected ITS length variants by PCR-RFLP. No variation at the number of Ag-NORs per metaphase was found among the 51 durum wheat cultivars, but the PCR-RFLP technique carried out with the restriction enzyme HpaII, allowed the detection of ITS length variants among them. The molecular data was used in order to establish the genetic relationships among cultivars and botanical varieties of durum wheat. The knowledge of this feature could be useful for future design of breeding strategies, involving this collection that constitutes an excellent repository of germplasm in Portugal.

The intergenic spacer of the ribosomal DNA is highly variable, but is location specific in the nucleolar organizer region of the chromosomes. This study provides an event of high level of polymorphism / size variation and occurrence of 14 unique phenotypes in 48 landraces of Portuguese bread wheat cultivars for IGS-amplified products obtained by PCR-RFLP technique performed with TaqI. The attendant IGS polymorphism has been used to deduce affinities between landraces. Some of the high molecular weight IGS allelic variants were also probed for their chromosomal localization by sequential silver nitrate staining and fluorescence in situ hybridization. However, only the intergenic spacer allelic variant of 3.1 kb could be successfully hybridized, and was observed to be physically located on the chromosome pair 1B in the NOR loci of the cultivar ‘Magueija’.