Flow sorting of C-genome chromosomes from wild relatives of wheat Aegilops markgrafii, Ae. triuncialis and Ae. cylindrica, and their molecular organization

Molnár, István and Vrána, Jan and Farkas, András and Kubaláková, Marie and Cseh, András Attila and Lángné Molnár, Márta (2015) Flow sorting of C-genome chromosomes from wild relatives of wheat Aegilops markgrafii, Ae. triuncialis and Ae. cylindrica, and their molecular organization. ANNALS OF BOTANY, 116 (2). pp. 189-200. ISSN 0305-7364

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Abstract

Background and Aims Aegilops caudata (CC) and its natural hybrids Ae. triuncialis (U t U t C t C t ) and Ae. cylindrica (D c D c C c C c ) represent a rich reservoir of useful genes for wheat improvement, but the limited information available on their gen ome structure and the shortage of molecular (cyto-) genetic tools hamper the utilization of th e extant genetic diversity. This study provides the complete karyotypes obtained after FISH with repetitive DNA probes, and evaluates the potential of flow cytometric chromosome sorting i n the three species. • Methods The flow karyotypes obtained after the analysis of DAPI-stained chromosomes were characterized and the chromosome content of the peaks on t he flow karyotypes was determined by FISH. Twenty-nine conserved orthologous set (COS) markers covering all seven wheat homoeologous chromosome groups were used for PCR wi th DNA amplified from flow-sorted chromosomes and genomic DNA. • Key Results FISH with repetitive DNA probes revealed that chromosomes 4C, 5 C, 7C t , T6U t S.6U t L-5C t L, 1C c and 5D c could be sorted with purities ranging from 66 to 91%, while the remaining chromosomes could be sorted in groups of two to f ive. This identified a partial wheat–C genome homology for group 4 and 5 chromosomes. In addit ion, 1C chromosomes were homologous with group 1 of wheat; a small segment from gro up 2 indicated 1C-2C rearrangement. The extensively rearranged structure of chromoso me 7C relative to wheat was also detected.Conclusions The possibility of purifying Aegilops chromosomes provides an attractive opportunity to investigate the structure and evolution of the Aegilops C genome and to develop molecular tools to facilitate the identification of ali en chromatin and support alien introgression breeding in wheat.

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