Genome Survey of the Freshwater Mussel Venustaconcha ellipsiformis (Bivalvia: Unionida) Using a Hybrid De Novo Assembly Approach

Freshwater mussels (Bivalvia: Unionida) serve an important role as aquatic ecosystem engineers but are one of the most critically imperilled groups of animals. Here, we used a combination of sequencing strategies to assemble and annotate a draft genome of Venustaconcha ellipsiformis, which will serve as a valuable genomic resource given the ecological value and unique “doubly uniparental inheritance” mode of mitochondrial DNA transmission of freshwater mussels. The genome described here was obtained by combining high coverage short reads (65X genome coverage of Illumina paired-end and 11X genome coverage of mate-pairs sequences) with low coverage Pacific Biosciences long reads (0.3X genome coverage). Briefly, the final scaffold assembly accounted for a total size of 1.54Gb (366,926 scaffolds, N50 = 6.5Kb, with 2.3% of “N” nucleotides), representing 86% of the predicted genome size of 1.80Gb, while over one third of the genome (37.5%) consisted of repeated elements and more than 85% of the core eukaryotic genes were recovered. Given the repeated genetic bottlenecks of V. ellipsiformis populations as a result of glaciations events, heterozygosity was also found to be remarkably low (0.6%), in contrast to most other sequenced bivalve species. Finally, we reassembled the full mitochondrial genome and found six polymorphic sites with respect to the previously published reference. This resource opens the way to comparative genomics studies to identify genes related to the unique adaptations of freshwater mussels and their distinctive mitochondrial inheritance mechanism.

[1]  Nathan A. Johnson,et al.  Deciphering the Link between Doubly Uniparental Inheritance of mtDNA and Sex Determination in Bivalves: Clues from Comparative Transcriptomics , 2018, Genome biology and evolution.

[2]  C. Mazzoni,et al.  A hybrid-hierarchical genome assembly strategy to sequence the invasive golden mussel, Limnoperna fortunei , 2017, GigaScience.

[3]  M. Seddon,et al.  Conservation of freshwater bivalves at the global scale: diversity, threats and research needs , 2018, Hydrobiologia.

[4]  Ruiqiang Li,et al.  Scallop genome reveals molecular adaptations to semi-sessile life and neurotoxins , 2017, Nature Communications.

[5]  Michael C. Schatz,et al.  Hybrid assembly with long and short reads improves discovery of gene family expansions , 2017, BMC Genomics.

[6]  Han Fang,et al.  GenomeScope: Fast reference-free genome profiling from short reads , 2016, bioRxiv.

[7]  L. Rieseberg,et al.  The sunflower genome provides insights into oil metabolism, flowering and Asterid evolution , 2017, Nature.

[8]  Kyudong Han,et al.  The Whole-Genome and Transcriptome of the Manila Clam (Ruditapes philippinarum) , 2017, Genome biology and evolution.

[9]  Justin Chu,et al.  ABySS 2.0: resource-efficient assembly of large genomes using a Bloom filter , 2016, bioRxiv.

[10]  Yanjie Zhang,et al.  Adaptation to deep-sea chemosynthetic environments as revealed by mussel genomes , 2017, Nature Ecology &Evolution.

[11]  Ruiqiang Li,et al.  Scallop genome provides insights into evolution of bilaterian karyotype and development , 2017, Nature Ecology &Evolution.

[12]  V. Simić,et al.  Conservation status of freshwater mussels in Europe: state of the art and future challenges , 2017, Biological reviews of the Cambridge Philosophical Society.

[13]  J. Corbeil,et al.  Draft genome of the American Eel (Anguilla rostrata) , 2014, Molecular ecology resources.

[14]  D. Stewartb,et al.  Sex Determining Mechanisms in Bivalves , 2017 .

[15]  David Haussler,et al.  Long-read sequence assembly of the gorilla genome , 2016, Science.

[16]  D. Puiu,et al.  A First Insight into the Genome of the Filter-Feeder Mussel Mytilus galloprovincialis , 2016, PloS one.

[17]  Evgeny M. Zdobnov,et al.  BUSCO: assessing genome assembly and annotation completeness with single-copy orthologs , 2015, Bioinform..

[18]  S. Schaeffer,et al.  Species Distribution and Population Connectivity of Deep-Sea Mussels at Hydrocarbon Seeps in the Gulf of Mexico , 2015, PloS one.

[19]  M. Czarnoleski,et al.  Factors affecting trematode infection rates in freshwater mussels , 2014, Hydrobiologia.

[20]  S. Varandas,et al.  Biology and conservation of freshwater bivalves: past, present and future perspectives , 2014, Hydrobiologia.

[21]  Björn Usadel,et al.  Trimmomatic: a flexible trimmer for Illumina sequence data , 2014, Bioinform..

[22]  Paul Medvedev,et al.  Informed and automated k-mer size selection for genome assembly , 2013, Bioinform..

[23]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[24]  M. Lascoux,et al.  Ecological genomics of local adaptation , 2013, Nature Reviews Genetics.

[25]  A. T. Harris,et al.  Phylogeography and Genetic Variability of the Freshwater Mussels (Bivalvia: Unionidae) Ellipse, Venustaconcha ellipsiformis (Conrad 1836), and Bleeding Tooth, V. Pleasii (Marsh 1891) , 2013 .

[26]  Alexey A. Gurevich,et al.  QUAST: quality assessment tool for genome assemblies , 2013, Bioinform..

[27]  Jian Wang,et al.  SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler , 2012, GigaScience.

[28]  Qiang Wang,et al.  The oyster genome reveals stress adaptation and complexity of shell formation , 2012, Nature.

[29]  W. Haag North American Freshwater Mussels: Natural History, Ecology, and Conservation , 2012 .

[30]  E. Zouros Biparental Inheritance Through Uniparental Transmission: The Doubly Uniparental Inheritance (DUI) of Mitochondrial DNA , 2012, Evolutionary Biology.

[31]  M. Schatz,et al.  Hybrid error correction and de novo assembly of single-molecule sequencing reads , 2012, Nature Biotechnology.

[32]  Hideo Aoki,et al.  Draft Genome of the Pearl Oyster Pinctada fucata: A Platform for Understanding Bivalve Biology , 2012, DNA research : an international journal for rapid publication of reports on genes and genomes.

[33]  E. Chapman,et al.  Novel protein genes in animal mtDNA: a new sex determination system in freshwater mussels (Bivalvia: Unionoida)? , 2011, Molecular biology and evolution.

[34]  Carl Kingsford,et al.  A fast, lock-free approach for efficient parallel counting of occurrences of k-mers , 2011, Bioinform..

[35]  A. Rypel,et al.  Growth and longevity in freshwater mussels: evolutionary and conservation implications , 2011, Biological reviews of the Cambridge Philosophical Society.

[36]  A. Gnirke,et al.  High-quality draft assemblies of mammalian genomes from massively parallel sequence data , 2010, Proceedings of the National Academy of Sciences.

[37]  Aaron R. Quinlan,et al.  Bioinformatics Applications Note Genome Analysis Bedtools: a Flexible Suite of Utilities for Comparing Genomic Features , 2022 .

[38]  P. Blier,et al.  Comparative Mitochondrial Genomics of Freshwater Mussels (Bivalvia: Unionoida) With Doubly Uniparental Inheritance of mtDNA: Gender-Specific Open Reading Frames and Putative Origins of Replication , 2009, Genetics.

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

[40]  Steven J. M. Jones,et al.  Abyss: a Parallel Assembler for Short Read Sequence Data Material Supplemental Open Access , 2022 .

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

[42]  M. Passamonti,et al.  Doubly uniparental inheritance: two mitochondrial genomes, one precious model for organelle DNA inheritance and evolution. , 2009, DNA and cell biology.

[43]  D. C. Aldridge,et al.  Freshwater mussel abundance predicts biodiversity in UK lowland rivers , 2007 .

[44]  P. Blier,et al.  The unusual system of doubly uniparental inheritance of mtDNA: isn't one enough? , 2007, Trends in genetics : TIG.

[45]  C. Vaughn,et al.  Context-dependent effects of freshwater mussels on stream benthic communities , 2006 .

[46]  Steven Salzberg,et al.  TigrScan and GlimmerHMM: two open source ab initio eukaryotic gene-finders , 2004, Bioinform..

[47]  David L. Strayer,et al.  Mollusks as ecosystem engineers: the role of shell production in aquatic habitats , 2003 .

[48]  I. Longden,et al.  EMBOSS: the European Molecular Biology Open Software Suite. , 2000, Trends in genetics : TIG.

[49]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[50]  Burton H. Bloom,et al.  Space/time trade-offs in hash coding with allowable errors , 1970, CACM.