A chromosome-scale assembly of the sorghum genome using nanopore sequencing and optical mapping

The advent of long-read sequencing technologies has greatly facilitated assemblies of large eukaryotic genomes. In this paper, Oxford Nanopore sequences generated on a MinION sequencer were combined with BioNano Genomics Direct Label and Stain (DLS) optical maps to generate a chromosome-scale de novo assembly of the repeat-rich Sorghum bicolor Tx430 genome. The final hybrid assembly consists of 29 scaffolds, encompassing in most cases entire chromosome arms. It has a scaffold N50 value of 33.28Mbps and covers >90% of Sorghum bicolor expected genome length. A sequence accuracy of 99.67% was obtained in unique regions after aligning contigs against Illumina Tx430 data. Alignments showed that 99.4% of the 34,211 public gene models are present in the assembly, including 94.2% mapping end-to-end. Comparisons of the DLS optical maps against the public Sorghum Bicolor v3.0.1 BTx623 genome assembly suggest the presence of substantial genomic rearrangements whose origin remains to be determined.

[1]  Bernardo J. Clavijo,et al.  Improving and correcting the contiguity of long-read genome assemblies of three plant species using optical mapping and chromosome conformation capture data. , 2017, Genome research.

[2]  Christina A. Cuomo,et al.  Pilon: An Integrated Tool for Comprehensive Microbial Variant Detection and Genome Assembly Improvement , 2014, PloS one.

[3]  Dawn H. Nagel,et al.  The B73 Maize Genome: Complexity, Diversity, and Dynamics , 2009, Science.

[4]  T. Sakurai,et al.  Genome sequence of the palaeopolyploid soybean , 2010, Nature.

[5]  S. Oliver,et al.  Estimating the total number of phosphoproteins and phosphorylation sites in eukaryotic proteomes , 2017, GigaScience.

[6]  Ryan R. Wick,et al.  Completing bacterial genome assemblies with multiplex MinION sequencing , 2017, bioRxiv.

[7]  Stefan Engelen,et al.  de novo assembly and population genomic survey of natural yeast isolates with the Oxford Nanopore MinION sequencer , 2016, bioRxiv.

[8]  Jeffry D Sander,et al.  Developing a flexible, high‐efficiency Agrobacterium‐mediated sorghum transformation system with broad application , 2018, Plant biotechnology journal.

[9]  Jeffrey Ross-Ibarra,et al.  Improved maize reference genome with single-molecule technologies , 2017, Nature.

[10]  E. Pennisi New technologies boost genome quality. , 2017, Science.

[11]  Ute Roessner,et al.  The genome of Chenopodium quinoa , 2017, Nature.

[12]  S. Koren,et al.  Canu: scalable and accurate long-read assembly via adaptive k-mer weighting and repeat separation , 2016, bioRxiv.

[13]  Heng Li,et al.  Minimap and miniasm: fast mapping and de novo assembly for noisy long sequences , 2015, Bioinform..

[14]  Mihaela M. Martis,et al.  The Sorghum bicolor genome and the diversification of grasses , 2009, Nature.

[15]  I. Godwin,et al.  Highly efficient sorghum transformation , 2012, Plant Cell Reports.

[16]  R. Wing,et al.  An improved method for plant BAC library construction. , 2003, Methods in molecular biology.

[17]  S. Koren,et al.  Nanopore sequencing and assembly of a human genome with ultra-long reads , 2017, bioRxiv.

[18]  S. Salzberg,et al.  Using MUMmer to Identify Similar Regions in Large Sequence Sets , 2003, Current protocols in bioinformatics.

[19]  Mick Watson,et al.  A single chromosome assembly of Bacteroides fragilis strain BE1 from Illumina and MinION nanopore sequencing data , 2015, GigaScience.

[20]  L. Mao,et al.  The Aegilops tauschii genome reveals multiple impacts of transposons , 2017, Nature Plants.

[21]  Detlef Weigel,et al.  High contiguity Arabidopsis thaliana genome assembly with a single nanopore flow cell , 2018, Nature Communications.

[22]  Karl G. Kugler,et al.  Genome sequence of the progenitor of the wheat D genome Aegilops tauschii , 2017, Nature.

[23]  N. Loman,et al.  A complete bacterial genome assembled de novo using only nanopore sequencing data , 2015, Nature Methods.

[24]  Steven L Salzberg,et al.  Fast gapped-read alignment with Bowtie 2 , 2012, Nature Methods.

[25]  Sergey Koren,et al.  De Novo Assembly of a New Solanum pennellii Accession Using Nanopore Sequencing[CC-BY] , 2017, Plant Cell.

[26]  Haibao Tang,et al.  Single-molecule sequencing of the desiccation-tolerant grass Oropetium thomaeum , 2015, Nature.