Wheat Neocentromeres Found in F1 Triticale × Tritordeum Hybrids (AABBRHch) After 5-Azacytidine Treatment

  • A. Carvalho
  • H. Guedes-Pinto
  • J. S. Heslop-Harrison
  • J. Lima-BritoEmail author


The DNA hypomethylation effect of 5-azacytine (5-AC; a cytosine analog) is widely known. This agent has been used for rRNA gene expression studies of Triticeae amphiploids and hybrids regarding rye rRNA genes suppression caused by the wheat nucleolar dominance phenomenon. However, this situation is reverted by 5-AC treatment which activates rye rRNA gene expression as it has been intensively observed in triticale. For nucleolar dominance studies, we produced F1 multigeneric hybrids (AABBRHch; 2n = 6x = 42) from crosses between the triticale cultivar ‘Corgo’ (AABBRR; 2n = 6x = 42) and the tritordeum cultivars HT9 and HT31 (AABBHchHch; 2n = 6x = 42). The hybrid seeds were germinated in a low concentration of 5-AC (treatment) and in distilled water (nontreated control plants). Silver nitrate staining performed in one 5-AC-treated F1 hybrid revealed a reduced number of interphase cells with seven nucleoli, metaphases with eight Ag-NORs, and neocentromeres in the long arm of three wheat chromosomes. Nontreated hybrids presented six Ag-NORs per mitotic metaphase cell and a maximum of six nucleoli per interphase because of the 1R Ag-NOR suppression. No neocentromere was found in the control F1 hybrid plants. Both treated and nontreated seedlings were subsequently evaluated by fluorescent in situ hybridization performed with genomic and repetitive DNA probes to identify Hch and rye genomes, to confirm Ag-NORs location, and to detect inactive rDNA loci. DAPI counterstaining was also helpful for the detection of neocentromeres in the long arm of three wheat chromosomes. This study allowed us to suggest that 5-AC treatment specifically induced wheat neocentromeres in the F1 multigeneric triticale × tritordeum hybrids.


5-Azacytidine FISH Neocentromeres NOR Triticum aestivum L. 





fluorescent in situ hybridization


ribosomal DNA



This work was supported by the POCTI/35107/AGR/2000 and PTDC/AGR-GPL/65876/2006 projects and the Ph.D. grant SFRH/BD/17348/2004 financed by the Portuguese Foundation for Science and the Technology (FCT).


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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • A. Carvalho
    • 1
  • H. Guedes-Pinto
    • 1
  • J. S. Heslop-Harrison
    • 2
  • J. Lima-Brito
    • 1
    Email author
  1. 1.Institute for Biotechnology and Bioengineering, Centre of Genetics and Biotechnology (IBB/CGB)University of Trás-os-Montes and Alto Douro (UTAD)Vila RealPortugal
  2. 2.Department of BiologyUniversity of LeicesterLeicesterUK

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