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The genome of flax (Linum usitatissimum) assembled de novo from short shotgun sequence reads
Authors:Zhiwen Wang  Neil Hobson  Leonardo Galindo  Shilin Zhu  Daihu Shi  Joshua McDill  Linfeng Yang  Simon Hawkins  Godfrey Neutelings  Raju Datla  Georgina Lambert  David W. Galbraith  Christopher J. Grassa  Armando Geraldes  Quentin C. Cronk  Christopher Cullis  Prasanta K. Dash  Polumetla A. Kumar  Sylvie Cloutier  Andrew G. Sharpe  Gane K.‐S. Wong  Jun Wang  Michael K. Deyholos
Affiliation:1. BGI‐Shenzen, Bei Shan Industrial Zone, Yantian District, Shenzhen 518083, China;2. Department of Biological Sciences, University of Alberta, Edmonton, Alberta, T6G 2E9, Canada;3. Université Lille‐Nord de France, Lille 1 Unité Mixte de Recherche Institut National de la Recherche Agronomique 1281, Stress Abiotiques et Différenciation des Végétaux, F‐59650 Villeneuve d’Ascq Cedex, France;4. National Research Council of Canada, Plant Biotechnology Institute, Saskatoon, Saskatchewan, S7N 0W9, Canada;5. University of Arizona, School of Plant Sciences and BIO5 Institute, Tucson, AZ 85721, USA;6. Department of Botany, University of British Columbia, Vancouver, British Columbia, V6T 1Z4, Canada;7. Case Western Reserve University, Cleveland, OH 44106, USA;8. National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute, Pusa Campus, New Delhi 110012, India;9. Agriculture and Agri‐Food Canada, 195 Dafoe Road, Winnipeg, Manitoba, R3T 2M1, Canada;10. Department of Plant Science, University of Manitoba, Winnipeg, Manitoba, R3T 2N2, Canada;11. Department of Medicine, University of Alberta, Edmonton, Alberta, T6G 2E1, Canada;12. The Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Denmark;13. Department of Biology, University of Copenhagen, Denmark
Abstract:Flax (Linum usitatissimum) is an ancient crop that is widely cultivated as a source of fiber, oil and medicinally relevant compounds. To accelerate crop improvement, we performed whole‐genome shotgun sequencing of the nuclear genome of flax. Seven paired‐end libraries ranging in size from 300 bp to 10 kb were sequenced using an Illumina genome analyzer. A de novo assembly, comprised exclusively of deep‐coverage (approximately 94× raw, approximately 69× filtered) short‐sequence reads (44–100 bp), produced a set of scaffolds with N50 = 694 kb, including contigs with N50 = 20.1 kb. The contig assembly contained 302 Mb of non‐redundant sequence representing an estimated 81% genome coverage. Up to 96% of published flax ESTs aligned to the whole‐genome shotgun scaffolds. However, comparisons with independently sequenced BACs and fosmids showed some mis‐assembly of regions at the genome scale. A total of 43 384 protein‐coding genes were predicted in the whole‐genome shotgun assembly, and up to 93% of published flax ESTs, and 86% of A. thaliana genes aligned to these predicted genes, indicating excellent coverage and accuracy at the gene level. Analysis of the synonymous substitution rates (Ks) observed within duplicate gene pairs was consistent with a recent (5–9 MYA) whole‐genome duplication in flax. Within the predicted proteome, we observed enrichment of many conserved domains (Pfam‐A) that may contribute to the unique properties of this crop, including agglutinin proteins. Together these results show that de novo assembly, based solely on whole‐genome shotgun short‐sequence reads, is an efficient means of obtaining nearly complete genome sequence information for some plant species.
Keywords:whole‐genome shotgun  DNA sequencing  Illumina  flax  Malpighiales  industrial crops
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