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Improvement of the banana “Musa acuminata” reference sequence using NGS data and semi-automated bioinformatics methods

Identifieur interne : 001C94 ( Main/Curation ); précédent : 001C93; suivant : 001C95

Improvement of the banana “Musa acuminata” reference sequence using NGS data and semi-automated bioinformatics methods

Auteurs : Guillaume Martin ; Franc-Christophe Baurens ; Gaëtan Droc ; Mathieu Rouard ; Alberto Cenci ; Andrzej Kilian ; Alex Hastie [États-Unis] ; Jaroslav Doležel [République tchèque] ; Jean-Marc Aury [France] ; Adriana Alberti [France] ; Françoise Carreel ; Angélique D Ont

Source :

RBID : PMC:4793746

Descripteurs français

English descriptors

Abstract

Background

Recent advances in genomics indicate functional significance of a majority of genome sequences and their long range interactions. As a detailed examination of genome organization and function requires very high quality genome sequence, the objective of this study was to improve reference genome assembly of banana (Musa acuminata).

Results

We have developed a modular bioinformatics pipeline to improve genome sequence assemblies, which can handle various types of data. The pipeline comprises several semi-automated tools. However, unlike classical automated tools that are based on global parameters, the semi-automated tools proposed an expert mode for a user who can decide on suggested improvements through local compromises. The pipeline was used to improve the draft genome sequence of Musa acuminata. Genotyping by sequencing (GBS) of a segregating population and paired-end sequencing were used to detect and correct scaffold misassemblies. Long insert size paired-end reads identified scaffold junctions and fusions missed by automated assembly methods. GBS markers were used to anchor scaffolds to pseudo-molecules with a new bioinformatics approach that avoids the tedious step of marker ordering during genetic map construction. Furthermore, a genome map was constructed and used to assemble scaffolds into super scaffolds. Finally, a consensus gene annotation was projected on the new assembly from two pre-existing annotations. This approach reduced the total Musa scaffold number from 7513 to 1532 (i.e. by 80 %), with an N50 that increased from 1.3 Mb (65 scaffolds) to 3.0 Mb (26 scaffolds). 89.5 % of the assembly was anchored to the 11 Musa chromosomes compared to the previous 70 %. Unknown sites (N) were reduced from 17.3 to 10.0 %.

Conclusion

The release of the Musa acuminata reference genome version 2 provides a platform for detailed analysis of banana genome variation, function and evolution. Bioinformatics tools developed in this work can be used to improve genome sequence assemblies in other species.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-016-2579-4) contains supplementary material, which is available to authorized users.


Url:
DOI: 10.1186/s12864-016-2579-4
PubMed: 26984673
PubMed Central: 4793746

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PMC:4793746

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Guillaume Martin
<affiliation>
<nlm:aff id="Aff1">CIRAD (Centre de coopération Internationale en Recherche Agronomique pour le Développement), UMR AGAP, TA A-108/03, Avenue Agropolis, F-34398, Montpellier, cedex 5 France</nlm:aff>
<wicri:noCountry code="subfield">cedex 5 France</wicri:noCountry>
</affiliation>
Franc-Christophe Baurens
<affiliation>
<nlm:aff id="Aff1">CIRAD (Centre de coopération Internationale en Recherche Agronomique pour le Développement), UMR AGAP, TA A-108/03, Avenue Agropolis, F-34398, Montpellier, cedex 5 France</nlm:aff>
<wicri:noCountry code="subfield">cedex 5 France</wicri:noCountry>
</affiliation>
Gaëtan Droc
<affiliation>
<nlm:aff id="Aff1">CIRAD (Centre de coopération Internationale en Recherche Agronomique pour le Développement), UMR AGAP, TA A-108/03, Avenue Agropolis, F-34398, Montpellier, cedex 5 France</nlm:aff>
<wicri:noCountry code="subfield">cedex 5 France</wicri:noCountry>
</affiliation>
Mathieu Rouard
<affiliation>
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</affiliation>
Alberto Cenci
<affiliation>
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<wicri:noCountry code="subfield">Cedex 5 France</wicri:noCountry>
</affiliation>
Andrzej Kilian
<affiliation>
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<wicri:noCountry code="subfield">Australian Capital Territory 2600 Australia</wicri:noCountry>
</affiliation>
Françoise Carreel
<affiliation>
<nlm:aff id="Aff1">CIRAD (Centre de coopération Internationale en Recherche Agronomique pour le Développement), UMR AGAP, TA A-108/03, Avenue Agropolis, F-34398, Montpellier, cedex 5 France</nlm:aff>
<wicri:noCountry code="subfield">cedex 5 France</wicri:noCountry>
</affiliation>
Angélique D Ont
<affiliation>
<nlm:aff id="Aff1">CIRAD (Centre de coopération Internationale en Recherche Agronomique pour le Développement), UMR AGAP, TA A-108/03, Avenue Agropolis, F-34398, Montpellier, cedex 5 France</nlm:aff>
<wicri:noCountry code="subfield">cedex 5 France</wicri:noCountry>
</affiliation>

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<name sortKey="Martin, Guillaume" sort="Martin, Guillaume" uniqKey="Martin G" first="Guillaume" last="Martin">Guillaume Martin</name>
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<name sortKey="Dolezel, Jaroslav" sort="Dolezel, Jaroslav" uniqKey="Dolezel J" first="Jaroslav" last="Doležel">Jaroslav Doležel</name>
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<term>Computational Biology (methods)</term>
<term>Contig Mapping</term>
<term>Genetic Markers</term>
<term>Genome, Plant</term>
<term>High-Throughput Nucleotide Sequencing</term>
<term>Molecular Sequence Annotation</term>
<term>Musa (genetics)</term>
<term>Sequence Analysis, DNA</term>
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<keywords scheme="KwdFr" xml:lang="fr">
<term>Analyse de séquence d'ADN</term>
<term>Annotation de séquence moléculaire</term>
<term>Biologie informatique ()</term>
<term>Cartographie de contigs</term>
<term>Génome végétal</term>
<term>Marqueurs génétiques</term>
<term>Musa (génétique)</term>
<term>Séquençage nucléotidique à haut débit</term>
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<term>Genetic Markers</term>
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<term>Musa</term>
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<term>Computational Biology</term>
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<keywords scheme="MESH" xml:lang="en">
<term>Contig Mapping</term>
<term>Genome, Plant</term>
<term>High-Throughput Nucleotide Sequencing</term>
<term>Molecular Sequence Annotation</term>
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<term>Analyse de séquence d'ADN</term>
<term>Annotation de séquence moléculaire</term>
<term>Biologie informatique</term>
<term>Cartographie de contigs</term>
<term>Génome végétal</term>
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<sec>
<title>Background</title>
<p>Recent advances in genomics indicate functional significance of a majority of genome sequences and their long range interactions. As a detailed examination of genome organization and function requires very high quality genome sequence, the objective of this study was to improve reference genome assembly of banana (
<italic>Musa acuminata</italic>
).</p>
</sec>
<sec>
<title>Results</title>
<p>We have developed a modular bioinformatics pipeline to improve genome sequence assemblies, which can handle various types of data. The pipeline comprises several semi-automated tools. However, unlike classical automated tools that are based on global parameters, the semi-automated tools proposed an expert mode for a user who can decide on suggested improvements through local compromises. The pipeline was used to improve the draft genome sequence of
<italic>Musa acuminata.</italic>
Genotyping by sequencing (GBS) of a segregating population and paired-end sequencing were used to detect and correct scaffold misassemblies. Long insert size paired-end reads identified scaffold junctions and fusions missed by automated assembly methods. GBS markers were used to anchor scaffolds to pseudo-molecules with a new bioinformatics approach that avoids the tedious step of marker ordering during genetic map construction. Furthermore, a genome map was constructed and used to assemble scaffolds into super scaffolds. Finally, a consensus gene annotation was projected on the new assembly from two pre-existing annotations. This approach reduced the total
<italic>Musa</italic>
scaffold number from 7513 to 1532 (i.e. by 80 %), with an N50 that increased from 1.3 Mb (65 scaffolds) to 3.0 Mb (26 scaffolds). 89.5 % of the assembly was anchored to the 11
<italic>Musa</italic>
chromosomes compared to the previous 70 %. Unknown sites (N) were reduced from 17.3 to 10.0 %.</p>
</sec>
<sec>
<title>Conclusion</title>
<p>The release of the
<italic>Musa acuminata</italic>
reference genome version 2 provides a platform for detailed analysis of banana genome variation, function and evolution. Bioinformatics tools developed in this work can be used to improve genome sequence assemblies in other species.</p>
</sec>
<sec>
<title>Electronic supplementary material</title>
<p>The online version of this article (doi:10.1186/s12864-016-2579-4) contains supplementary material, which is available to authorized users.</p>
</sec>
</div>
</front>
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<name sortKey="Baurens, F C" uniqKey="Baurens F">F-C Baurens</name>
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<name sortKey="Cenci, A" uniqKey="Cenci A">A Cenci</name>
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<name sortKey="Ruiz, M" uniqKey="Ruiz M">M Ruiz</name>
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