Čeština
EnglishChromosome sorting and sequencing facilitate cytogenetic mapping of satellite repeat landscape of the U- and M genomes of diploid Aegilops
Molnár I., Jurečka J., Said M., Farkas A., Ivanizs L., Doležel J. and Hřibová E.
Plant Biotechnology: Green for Good V : , 2019
Keywords:
Abstract: Aegilops umbellulata (2n=2x=14; UU) and Ae. comosa (2n=2x=14, MM) species are rich sources of useful genes and alleles for wheat improvement. Mapping satellite DNA repeats using fluorescence in situ hybridization (FISH) is a powerful method to identify Aegilops chromosomes and monitoring introgression of alien chromatin into wheat. In the present work, we used sequence data from flow-sorted Aegilops chromosomes to identify new satellite repeats and tested their suitability for identification of U- and M genome chromosomes by FISH. Sequencing DNA of flow-sorted chromosomes 1U - 7U and chromosome arms 2US and 2UL of Ae. umbellulata by illumina resulted in ~26 Gbp sequence data per chromosome. 316.6 Mbp/chromosome were assembled covering ~40% of the estimated U genome size. Graph-based clustering of 1,420,161 reads resulted in 34,745 clusters where the dominant repeats were different types of LTR retrotransposons: Ty3/gypsy elements (35%), Ty1/copia elements (9%) and putative non-annotated LTR retroelements (5%). Putative satellite repeats were identified based on graph layout and Dotter analysis. Tandem repeats were PCR-amplified using genomic DNA of Ae. umbellulata as template with specific oligo-primers and labelled with biotin. Five out of 12 satellite repeats showed characteristic signals on U- and M genome chromosomes of Ae. umbellulata and Ae. comosa. The repeats showed specific signals on one chromosome (CL199, CL70), on three chromosomes (CL174), or on six chromosomes (CL191) of the two species. Various combinations of the satellite repeats with probes for pSc119.2, Afa family and pTa71 repeats, which were used previously for karyotype analysis in Triticum and Aegilops, will facilitate cytogenetic characterization of wheat–Aegilops chromosome addition, substitution or translocation lines.
DOI:
Fulltext: contact IEB authors
IEB authors: Jaroslav Doležel, Eva Hřibová, Istvan Molnar, Mahmoud Said
Plant Biotechnology: Green for Good V : , 2019
Keywords:
Abstract: Aegilops umbellulata (2n=2x=14; UU) and Ae. comosa (2n=2x=14, MM) species are rich sources of useful genes and alleles for wheat improvement. Mapping satellite DNA repeats using fluorescence in situ hybridization (FISH) is a powerful method to identify Aegilops chromosomes and monitoring introgression of alien chromatin into wheat. In the present work, we used sequence data from flow-sorted Aegilops chromosomes to identify new satellite repeats and tested their suitability for identification of U- and M genome chromosomes by FISH. Sequencing DNA of flow-sorted chromosomes 1U - 7U and chromosome arms 2US and 2UL of Ae. umbellulata by illumina resulted in ~26 Gbp sequence data per chromosome. 316.6 Mbp/chromosome were assembled covering ~40% of the estimated U genome size. Graph-based clustering of 1,420,161 reads resulted in 34,745 clusters where the dominant repeats were different types of LTR retrotransposons: Ty3/gypsy elements (35%), Ty1/copia elements (9%) and putative non-annotated LTR retroelements (5%). Putative satellite repeats were identified based on graph layout and Dotter analysis. Tandem repeats were PCR-amplified using genomic DNA of Ae. umbellulata as template with specific oligo-primers and labelled with biotin. Five out of 12 satellite repeats showed characteristic signals on U- and M genome chromosomes of Ae. umbellulata and Ae. comosa. The repeats showed specific signals on one chromosome (CL199, CL70), on three chromosomes (CL174), or on six chromosomes (CL191) of the two species. Various combinations of the satellite repeats with probes for pSc119.2, Afa family and pTa71 repeats, which were used previously for karyotype analysis in Triticum and Aegilops, will facilitate cytogenetic characterization of wheat–Aegilops chromosome addition, substitution or translocation lines.
DOI:
Fulltext: contact IEB authors
IEB authors: Jaroslav Doležel, Eva Hřibová, Istvan Molnar, Mahmoud Said