Čeština
EnglishCharacterization and Dissecting the Complex Nuclear Genome of Crested Wheatgrass by Chromosome Flow Sorting
Said M., Kubaláková M., Karafiátová M., Molnár I., Čížková J., Cabrera A., Doležel J., Vrána J.
10th International Conference Analytical Cytometry : , 2019
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Abstract: Agropyron species are a potential source of beneficial traits for wheat improvement. Among them, Agropyron cristatum comprises a complex of diploid, tetraploid and hexaploid forms with the basic genome P, with some accessions carrying supernumerary B chromosomes (Bs). In this work, we applied flow cytometry to characterize and dissect the complex genome of A. cristatum into individual chromosomes to facilitate its analysis. Flow cytometric analysis classified 26 accessions as diploid, 51 accessions as tetraploid and 3 accessions as hexaploid. Mean nuclear 2C DNA content of diploid plants was estimated as 12.99 pg DNA (1C genome size = 6.352 Gbp). Flow karyotypes obtained after the analysis of DAPI-stained mitotic chromosomes of diploid and tetraploid accessions consisted of three peaks, each corresponding to a group of two or three chromosome types. To improve the resolution, bivariate flow karyotyping after fluorescent labeling of (GAA)7 microsatellite was applied and allowed discrimination and sorting of P genome chromosomes from wheat-A. cristatum addition lines. Chromosomes 1P - 6P and seven telomeric chromosomes could be sorted at purities ranging from 81.7 to 98.2% in disomics and from 44.8 to 87.3% in telosomics. Chromosome 7P was sorted at purities reaching 50.0 and 39.5% in diploid and tetraploid A. cristatum, respectively. In addition to the whole complement chromosomes (A), supernumerary B chromosomes (Bs) could be easily discriminated and sorted from a diploid accession at 95.4% purity. The sorted chromosomes will streamline genome analysis of A. cristatum, facilitating gene cloning and development of molecular tools to support alien introgression into wheat. ACKNOWLEDGEMENT Supported by GACR Grant 16-16992S • ERDF (No. CZ.02.1.01/0.0/0.0/16_019/0000827) • OTKA K116277 • MarieCurie AEGILWHEAT H2020-MSCA-IF-2016-746253 • Grants award AGL2014-52445-R and RTI2018-093367-B-I00 co-financed by FEDER (EU).
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IEB authors: Jana Čížková, Jaroslav Doležel, Miroslava Karafiá..., Istvan Molnar, Mahmoud Said, Jan Vrána
10th International Conference Analytical Cytometry : , 2019
Keywords:
Abstract: Agropyron species are a potential source of beneficial traits for wheat improvement. Among them, Agropyron cristatum comprises a complex of diploid, tetraploid and hexaploid forms with the basic genome P, with some accessions carrying supernumerary B chromosomes (Bs). In this work, we applied flow cytometry to characterize and dissect the complex genome of A. cristatum into individual chromosomes to facilitate its analysis. Flow cytometric analysis classified 26 accessions as diploid, 51 accessions as tetraploid and 3 accessions as hexaploid. Mean nuclear 2C DNA content of diploid plants was estimated as 12.99 pg DNA (1C genome size = 6.352 Gbp). Flow karyotypes obtained after the analysis of DAPI-stained mitotic chromosomes of diploid and tetraploid accessions consisted of three peaks, each corresponding to a group of two or three chromosome types. To improve the resolution, bivariate flow karyotyping after fluorescent labeling of (GAA)7 microsatellite was applied and allowed discrimination and sorting of P genome chromosomes from wheat-A. cristatum addition lines. Chromosomes 1P - 6P and seven telomeric chromosomes could be sorted at purities ranging from 81.7 to 98.2% in disomics and from 44.8 to 87.3% in telosomics. Chromosome 7P was sorted at purities reaching 50.0 and 39.5% in diploid and tetraploid A. cristatum, respectively. In addition to the whole complement chromosomes (A), supernumerary B chromosomes (Bs) could be easily discriminated and sorted from a diploid accession at 95.4% purity. The sorted chromosomes will streamline genome analysis of A. cristatum, facilitating gene cloning and development of molecular tools to support alien introgression into wheat. ACKNOWLEDGEMENT Supported by GACR Grant 16-16992S • ERDF (No. CZ.02.1.01/0.0/0.0/16_019/0000827) • OTKA K116277 • MarieCurie AEGILWHEAT H2020-MSCA-IF-2016-746253 • Grants award AGL2014-52445-R and RTI2018-093367-B-I00 co-financed by FEDER (EU).
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