High-Quality de Novo Genome Assembly of the Dekkera bruxellensis Yeast Using Nanopore MinION Sequencing

Genetic variation in natural populations represents the raw material for phenotypic diversity. Species-wide characterization of genetic variants is crucial to have a deeper insight into the genotype-phenotype relationship. With the advent of new sequencing strategies and more recently the release of long-read sequencing platforms, it is now possible to explore the genetic diversity of any nonmodel organisms, representing a fundamental resource for biological research. In the frame of population genomic surveys, a first step is to obtain the complete sequence and high-quality assembly of a reference genome. Here, we sequenced and assembled a reference genome of the nonconventional Dekkera bruxellensis yeast. While this species is a major cause of wine spoilage, it paradoxically contributes to the specific flavor profile of some Belgium beers. In addition, an extreme karyotype variability is observed across natural isolates, highlighting that D. bruxellensis genome is very dynamic. The whole genome of the D. bruxellensis UMY321 isolate was sequenced using a combination of Nanopore long-read and Illumina short-read sequencing data. We generated the most complete and contiguous de novo assembly of D. bruxellensis to date and obtained a first glimpse into the genomic variability within this species by comparing the sequences of several isolates. This genome sequence is therefore of high value for population genomic surveys and represents a reference to study genome dynamic in this yeast species.

[1]  Leonid Kruglyak,et al.  Comprehensive polymorphism survey elucidates population structure of Saccharomyces cerevisiae , 2009, Nature.

[2]  Walter Pirovano,et al.  SSPACE-LongRead: scaffolding bacterial draft genomes using long read sequence information , 2014, BMC Bioinformatics.

[3]  Keith Bradnam,et al.  CEGMA: a pipeline to accurately annotate core genes in eukaryotic genomes , 2007, Bioinform..

[4]  S. Salzberg,et al.  Versatile and open software for comparing large genomes , 2004, Genome Biology.

[5]  G. Fischer,et al.  Population Genomics Reveals Chromosome-Scale Heterogeneous Evolution in a Protoploid Yeast , 2014, Molecular biology and evolution.

[6]  Christian Brion,et al.  Evolution of intraspecific transcriptomic landscapes in yeasts , 2015, Nucleic acids research.

[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]  Pedro Almeida,et al.  Distinct Domestication Trajectories in Top-Fermenting Beer Yeasts and Wine Yeasts , 2016, Current Biology.

[9]  M. DePristo,et al.  The Genome Analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. , 2010, Genome research.

[10]  Robert P. Davey,et al.  Population genomics of domestic and wild yeasts , 2008, Nature.

[11]  Daniel A. Skelly,et al.  The 100-genomes strains, an S. cerevisiae resource that illuminates its natural phenotypic and genotypic variation and emergence as an opportunistic pathogen , 2015, Genome research.

[12]  K. Verstrepen,et al.  Assessing Genetic Diversity among Brettanomyces Yeasts by DNA Fingerprinting and Whole-Genome Sequencing , 2014, Applied and Environmental Microbiology.

[13]  Francesco Vezzi,et al.  De novo assembly of Dekkera bruxellensis: a multi technology approach using short and long-read sequencing and optical mapping , 2015, GigaScience.

[14]  Richard Durbin,et al.  Sequence analysis Fast and accurate short read alignment with Burrows – Wheeler transform , 2009 .

[15]  I. Pretorius,et al.  Genomic insights into the evolution of industrial yeast species Brettanomyces bruxellensis. , 2014, FEMS yeast research.

[16]  G. Bell,et al.  Speciation driven by hybridization and chromosomal plasticity in a wild yeast , 2015, Nature Microbiology.

[17]  Brendan D. O'Fallon,et al.  The genomic and phenotypic diversity of Schizosaccharomyces pombe , 2015, Nature Genetics.

[18]  Maitreya J. Dunham,et al.  Loss of Heterozygosity Drives Adaptation in Hybrid Yeast , 2017, Molecular biology and evolution.

[19]  Anthony R. Borneman,et al.  De-Novo Assembly and Analysis of the Heterozygous Triploid Genome of the Wine Spoilage Yeast Dekkera bruxellensis AWRI1499 , 2012, PloS one.

[20]  Pavel A. Pevzner,et al.  Assembly of long error-prone reads using de Bruijn graphs , 2016, Proceedings of the National Academy of Sciences.

[21]  J. Landolin,et al.  Assembling large genomes with single-molecule sequencing and locality-sensitive hashing , 2014, Nature Biotechnology.

[22]  Christian Brion,et al.  Differences in environmental stress response among yeasts is consistent with species-specific lifestyles , 2016, Molecular biology of the cell.

[23]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[24]  J. Piškur,et al.  The genome of wine yeast Dekkera bruxellensis provides a tool to explore its food-related properties. , 2012, International journal of food microbiology.

[25]  A. Couloux,et al.  A population genomics insight into the Mediterranean origins of wine yeast domestication , 2015, Molecular ecology.

[26]  J. Piškur,et al.  The wine and beer yeast Dekkera bruxellensis , 2014, Yeast.

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

[28]  D. Sherman,et al.  A Gondwanan imprint on global diversity and domestication of wine and cider yeast Saccharomyces uvarum , 2014, Nature Communications.

[29]  Christina A Cuomo,et al.  Genetic and phenotypic intra-species variation in Candida albicans , 2015, Genome research.

[30]  A. Borneman,et al.  Insights into the Dekkera bruxellensis Genomic Landscape: Comparative Genomics Reveals Variations in Ploidy and Nutrient Utilisation Potential amongst Wine Isolates , 2014, PLoS genetics.

[31]  J. Piškur,et al.  Complex Nature of the Genome in a Wine Spoilage Yeast, Dekkera bruxellensis , 2009, Eukaryotic Cell.

[32]  Paul M. Magwene,et al.  Outcrossing, mitotic recombination, and life-history trade-offs shape genome evolution in Saccharomyces cerevisiae , 2011, Proceedings of the National Academy of Sciences.

[33]  Joseph Schacherer,et al.  Population genomics of yeasts: towards a comprehensive view across a broad evolutionary scale , 2016, Yeast.

[34]  I. Masneuf-Pomarède,et al.  Development of microsatellite markers for the rapid and reliable genotyping of Brettanomyces bruxellensis at strain level. , 2014, Food microbiology.

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

[36]  William Stafford Noble,et al.  Integrative phenomics reveals insight into the structure of phenotypic diversity in budding yeast , 2013, Genome research.

[37]  T. C. White,et al.  The evolution of drug resistance in clinical isolates of Candida albicans , 2015, eLife.

[38]  K. Ohta,et al.  Population Genomics of the Fission Yeast Schizosaccharomyces pombe , 2014, PloS one.

[39]  I. Masneuf-Pomarède,et al.  The Genetics of Non-conventional Wine Yeasts: Current Knowledge and Future Challenges , 2016, Front. Microbiol..

[40]  Hugh E. Olsen,et al.  The Oxford Nanopore MinION: delivery of nanopore sequencing to the genomics community , 2016, Genome Biology.

[41]  Leopold Parts,et al.  A High-Definition View of Functional Genetic Variation from Natural Yeast Genomes , 2014, Molecular biology and evolution.

[42]  G. Sherlock,et al.  Whole Genome Analysis of 132 Clinical Saccharomyces cerevisiae Strains Reveals Extensive Ploidy Variation , 2016, G3: Genes, Genomes, Genetics.

[43]  Guy Baele,et al.  Domestication and Divergence of Saccharomyces cerevisiae Beer Yeasts , 2006, Cell.