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BioNano genome mapping of individual chromosomes supports physical mapping and sequence assembly in complex plant genomes
Authors:Helena Staňková  Alex R Hastie  Saki Chan  Jan Vrána  Zuzana Tulpová  Marie Kubaláková  Paul Visendi  Satomi Hayashi  Mingcheng Luo  Jacqueline Batley  David Edwards  Jaroslav Dole?el  Hana ?imková
Institution:1. Institute of Experimental Botany, Centre of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic;2. BioNano Genomics, San Diego, CA, USA;3. Australian Centre for Plant Functional Genomics, University of Queensland, Brisbane, QLD, Australia;4. School of Agriculture and Food Sciences, University of Queensland, Brisbane, QLD, Australia;5. Department of Plant Sciences, University of California, Davis, CA, USA;6. School of Plant Biology, University of Western Australia, Crawley, WA, Australia
Abstract:The assembly of a reference genome sequence of bread wheat is challenging due to its specific features such as the genome size of 17 Gbp, polyploid nature and prevalence of repetitive sequences. BAC‐by‐BAC sequencing based on chromosomal physical maps, adopted by the International Wheat Genome Sequencing Consortium as the key strategy, reduces problems caused by the genome complexity and polyploidy, but the repeat content still hampers the sequence assembly. Availability of a high‐resolution genomic map to guide sequence scaffolding and validate physical map and sequence assemblies would be highly beneficial to obtaining an accurate and complete genome sequence. Here, we chose the short arm of chromosome 7D (7DS) as a model to demonstrate for the first time that it is possible to couple chromosome flow sorting with genome mapping in nanochannel arrays and create a de novo genome map of a wheat chromosome. We constructed a high‐resolution chromosome map composed of 371 contigs with an N50 of 1.3 Mb. Long DNA molecules achieved by our approach facilitated chromosome‐scale analysis of repetitive sequences and revealed a ~800‐kb array of tandem repeats intractable to current DNA sequencing technologies. Anchoring 7DS sequence assemblies obtained by clone‐by‐clone sequencing to the 7DS genome map provided a valuable tool to improve the BAC‐contig physical map and validate sequence assembly on a chromosome‐arm scale. Our results indicate that creating genome maps for the whole wheat genome in a chromosome‐by‐chromosome manner is feasible and that they will be an affordable tool to support the production of improved pseudomolecules.
Keywords:optical mapping  wheat  sequencing  physical map  flow sorting  chromosomes
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