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      A reference genome for Nicotiana tabacum enables map-based cloning of homeologous loci implicated in nitrogen utilization efficiency

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          Abstract

          Background

          Tobacco ( Nicotiana tabacum) is an important plant model system that has played a key role in the early development of molecular plant biology. The tobacco genome is large and its characterisation challenging because it is an allotetraploid, likely arising from hybridisation between diploid N. sylvestris and N. tomentosiformis ancestors. A draft assembly was recently published for N. tabacum, but because of the aforementioned genome complexities it was of limited utility due to a high level of fragmentation.

          Results

          Here we report an improved tobacco genome assembly, which, aided by the application of optical mapping, achieves an N 50 size of 2.17 Mb and enables anchoring of 64% of the genome to pseudomolecules; a significant increase from the previous value of 19%. We use this assembly to identify two homeologous genes that explain the differentiation of the burley tobacco market class, with potential for greater understanding of Nitrogen Utilization Efficiency and Nitrogen Use Efficiency in plants; an important trait for future sustainability of agricultural production.

          Conclusions

          Development of an improved genome assembly for N. tabacum enables what we believe to be the first successful map-based gene discovery for the species, and demonstrates the value of an improved assembly for future research in this model and commercially-important species.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12864-017-3791-6) contains supplementary material, which is available to authorized users.

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          Most cited references47

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          Gene finding in novel genomes

          Ian Korf (2004)
          Background Computational gene prediction continues to be an important problem, especially for genomes with little experimental data. Results I introduce the SNAP gene finder which has been designed to be easily adaptable to a variety of genomes. In novel genomes without an appropriate gene finder, I demonstrate that employing a foreign gene finder can produce highly inaccurate results, and that the most compatible parameters may not come from the nearest phylogenetic neighbor. I find that foreign gene finders are more usefully employed to bootstrap parameter estimation and that the resulting parameters can be highly accurate. Conclusion Since gene prediction is sensitive to species-specific parameters, every genome needs a dedicated gene finder.
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            Toward almost closed genomes with GapFiller

            De novo assembly is a commonly used application of next-generation sequencing experiments. The ultimate goal is to puzzle millions of reads into one complete genome, although draft assemblies usually result in a number of gapped scaffold sequences. In this paper we propose an automated strategy, called GapFiller, to reliably close gaps within scaffolds using paired reads. The method shows good results on both bacterial and eukaryotic datasets, allowing only few errors. As a consequence, the amount of additional wetlab work needed to close a genome is drastically reduced. The software is available at http://www.baseclear.com/bioinformatics-tools/.
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              Reorganizing the protein space at the Universal Protein Resource (UniProt)

              The mission of UniProt is to support biological research by providing a freely accessible, stable, comprehensive, fully classified, richly and accurately annotated protein sequence knowledgebase, with extensive cross-references and querying interfaces. UniProt is comprised of four major components, each optimized for different uses: the UniProt Archive, the UniProt Knowledgebase, the UniProt Reference Clusters and the UniProt Metagenomic and Environmental Sequence Database. A key development at UniProt is the provision of complete, reference and representative proteomes. UniProt is updated and distributed every 4 weeks and can be accessed online for searches or download at http://www.uniprot.org.
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                Author and article information

                Contributors
                Kieron_edwards@bat.com , Kieron.edwards@cantab.net
                nf232@cornell.edu
                kedrake@ncsu.edu
                matthew_humphry@bat.com
                alan_evans@bat.com
                aurebg@vt.edu
                fraser_allen@bat.com
                robert_hurst@bat.com
                brian_white@bat.com
                sheri_kernodle@ncsu.edu
                Jennifer_Bromley@bat.com
                juan_sanchez@bat.com
                rslewis@ncsu.edu
                lam87@cornell.edu
                Journal
                BMC Genomics
                BMC Genomics
                BMC Genomics
                BioMed Central (London )
                1471-2164
                19 June 2017
                19 June 2017
                2017
                : 18
                : 448
                Affiliations
                [1 ]Plant Biotechnology Division, British American Tobacco, Cambridge, UK
                [2 ]ISNI 000000041936877X, GRID grid.5386.8, , Boyce Thompson Institute, ; Ithaca, NY USA
                [3 ]ISNI 0000 0001 2173 6074, GRID grid.40803.3f, Crop Science Department, , North Carolina State University, ; Raleigh, NC USA
                [4 ]ISNI 0000 0001 0694 4940, GRID grid.438526.e, , Present address Department of Horticulture, Virginia Tech, ; Blacksburg, VA USA
                Article
                3791
                10.1186/s12864-017-3791-6
                5474855
                28625162
                c58bc947-fee6-45f7-8bd1-afc8701b9d75
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 22 November 2016
                : 12 May 2017
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2017

                Genetics
                sequencing,nicotiana,nicotiana tabacum,tobacco,solanaceae,nitrogen use efficiency,nitrogen utilization efficiency,egy1,map-based cloning,polyploidy

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