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Reconstructing the Gigabase Plant Genome of Solanum pennellii using Nanopore Sequencing

Maximilian H.-W. Schmidt, Alxander Vogel, Alisandra K. Denton, Benjamin Istace, Alexandra Wormit, Henri van de Geest, Marie E. Bolger, Saleh Alseekh, Janina Maβ, Christian Pfaff, Ulrich Schurr, Roger Chetelat, Florian Maumus, Jean-Marc Aury, Alisdair R. Fernie, Dani Zamir, Anthony M. Bolger, Bjöern Usadel
doi: https://doi.org/10.1101/129148
Maximilian H.-W. Schmidt
1Institute for Botany and Molecular Genetics, BioEconomy Science Center, RWTH Aachen University, Aachen, Germany
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Alxander Vogel
1Institute for Botany and Molecular Genetics, BioEconomy Science Center, RWTH Aachen University, Aachen, Germany
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Alisandra K. Denton
1Institute for Botany and Molecular Genetics, BioEconomy Science Center, RWTH Aachen University, Aachen, Germany
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Benjamin Istace
2Commissariat à l’Energie Atomique et aux Energies Alternatives (CEA), Genoscope, 2 rue Gaston Crémieux, 91057 Evry, France
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Alexandra Wormit
1Institute for Botany and Molecular Genetics, BioEconomy Science Center, RWTH Aachen University, Aachen, Germany
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Henri van de Geest
3Wageningen Plant Research, Droevendaalsesteeg 1, 6708 PB, Wageningen, The Netherlands
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Marie E. Bolger
4Institute for Bio-and Geosciences (IBG-2: Plant Sciences), Forschungszentrum Jülich, Jülich, Germany
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Saleh Alseekh
5Department of Molecular Physiology, Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany
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Janina Maβ
4Institute for Bio-and Geosciences (IBG-2: Plant Sciences), Forschungszentrum Jülich, Jülich, Germany
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Christian Pfaff
4Institute for Bio-and Geosciences (IBG-2: Plant Sciences), Forschungszentrum Jülich, Jülich, Germany
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Ulrich Schurr
4Institute for Bio-and Geosciences (IBG-2: Plant Sciences), Forschungszentrum Jülich, Jülich, Germany
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Roger Chetelat
6C. M. Rick Tomato Genetics Resource Center, Department of Plant Sciences, University of California, Davis, California 95616
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Florian Maumus
7French National Institute for Agricultural Research (INRA), UR1164 Research Unit in Genomics Info (URGI), INRA de Versailles-Grignon, Versailles, France
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Jean-Marc Aury
2Commissariat à l’Energie Atomique et aux Energies Alternatives (CEA), Genoscope, 2 rue Gaston Crémieux, 91057 Evry, France
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Alisdair R. Fernie
5Department of Molecular Physiology, Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany
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Dani Zamir
8The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agriculture, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
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Anthony M. Bolger
1Institute for Botany and Molecular Genetics, BioEconomy Science Center, RWTH Aachen University, Aachen, Germany
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Bjöern Usadel
1Institute for Botany and Molecular Genetics, BioEconomy Science Center, RWTH Aachen University, Aachen, Germany
3Wageningen Plant Research, Droevendaalsesteeg 1, 6708 PB, Wageningen, The Netherlands
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  • For correspondence: usadel@bio1.rwth-aachen.de
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Abstract

Recent updates in sequencing technology have made it possible to obtain Gigabases of sequence data from one single flowcell. Prior to this update, the nanopore sequencing technology was mainly used to analyze and assemble microbial samples1-3. Here, we describe the generation of a comprehensive nanopore sequencing dataset with a median fragment size of 11,979 bp for the wild tomato species Solanum pennellii featuring an estimated genome size of ca 1.0 to 1.1 Gbases. We describe its genome assembly to a contig N50 of 2.5 MB using a pipeline comprising a Canu4 pre-processing and a subsequent assembly using SMARTdenovo. We show that the obtained nanopore based de novo genome reconstruction is structurally highly similar to that of the reference S. pennellii LA7165 genome but has a high error rate caused mostly by deletions in homopolymers. After polishing the assembly with Illumina short read data we obtained an error rate of <0.02 % when assessed versus the same Illumina data. More importantly however we obtained a gene completeness of 96.53% which even slightly surpasses that of the reference S. pennellii genome5. Taken together our data indicate such long read sequencing data can be used to affordably sequence and assemble Gbase sized diploid plant genomes.

Raw data is available at http://www.plabipd.de/portal/solanum-pennellii and has been deposited as PRJEB19787.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted April 21, 2017.
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Reconstructing the Gigabase Plant Genome of Solanum pennellii using Nanopore Sequencing
Maximilian H.-W. Schmidt, Alxander Vogel, Alisandra K. Denton, Benjamin Istace, Alexandra Wormit, Henri van de Geest, Marie E. Bolger, Saleh Alseekh, Janina Maβ, Christian Pfaff, Ulrich Schurr, Roger Chetelat, Florian Maumus, Jean-Marc Aury, Alisdair R. Fernie, Dani Zamir, Anthony M. Bolger, Bjöern Usadel
bioRxiv 129148; doi: https://doi.org/10.1101/129148
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Reconstructing the Gigabase Plant Genome of Solanum pennellii using Nanopore Sequencing
Maximilian H.-W. Schmidt, Alxander Vogel, Alisandra K. Denton, Benjamin Istace, Alexandra Wormit, Henri van de Geest, Marie E. Bolger, Saleh Alseekh, Janina Maβ, Christian Pfaff, Ulrich Schurr, Roger Chetelat, Florian Maumus, Jean-Marc Aury, Alisdair R. Fernie, Dani Zamir, Anthony M. Bolger, Bjöern Usadel
bioRxiv 129148; doi: https://doi.org/10.1101/129148

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