Biology subjects

Kocot, K.

Publications and source records attributed to Kocot, K..

6 recordsLinked to original sources

bioRxiv2026

The phylogenetic affinities of Chaetognathifera, with considerations of systematic error and the robusticity of macrosyntenic results

Chaetognathifera, a superphylum comprising Syndermata (Rotifera including Acanthocephala), Micrognathozoa, Gnathostomulida and Chaetognatha, is a complex grouping generally recovered as the sister to all other Lophotrochozoa. However, phylogenetic relationships within this group are controversial, in part due to poor sampling, resulting in two key questions. The first is whether Gnathostomulida or Chaetognatha represent the sister group to Syndermata+Micrognathozoa. The second is the phylogenetic position of the former phylum Acanthocephala within Syndermata. Here, we present the first study of the phylogenetic affinities of Chaetognathifera with genomic representation from all major phyla, and explore the potential of macrosynteny to better understand these relationships. For this latter aspect, we also developed a new jackknifing procedure to assess the robustness of linkage groups inferred by macrosyntenic analyses. We show that the phylogenetic relationships between these clades are corroborated through a variety of gene selection and analysis methodologies. This provides clear evidence of Acanthocephala as a derived clade within Syndermata as sister to Seisonidea, and that Gnathostomulida is sister to Syndermata+Micrognathozoa, with Chaetognatha as the earliest diverging clade within Chaetognathifera. On the other hand, we found that macrosyntenic patterns cannot resolve this question. Moreover, almost all possible linkage groups involving chaetognathiferan species lack robusticity and hence, should not be considered reliable. As a consequence so far, in Chaetognathifera none of the bilaterian ancestral linkage groups can be reliably found and independent massive chromosomal rearrangements occurred. We therefore strongly suggest that studies of macrosynteny should not only assess the significance of possible linkage groups, but also the robusticity of these linkage group inferences. Furthermore, we also present a script for this purpose, which can be found at: https://github.com/JFFleming/MacrosyntenicJackknife

evolutionary biology↗

bioRxiv2024

VEHoP: A Versatile, Easy-to-use, and Homology-based Phylogenomic pipeline accommodating diverse sequences

Phylogenomics has emerged as a transformative approach in systematics, conservation biology, and biomedicine, enabling the inference of evolutionary relationships by leveraging hundreds to thousands of genes from genomic or transcriptomic data. However, acquiring high-quality genomes and transcriptomes necessitates samples with intact DNA and RNA, substantial sequencing investments, and extensive bioinformatic processing, such as genome/transcriptome assembly and annotation. This challenge is particularly pronounced for rare or difficult-to-collect species, such as those inhabiting the deep sea, where only fragmented DNA reads are often available due to environmental degradation or suboptimal preservation conditions. To address these limitations, we introduce VEHoP (Versatile, Easy-to-use Homology-based Phylogenomic pipeline), a tool designed to infer protein-coding regions from diverse inputs, including raw reads (short and long), draft genomes, transcriptomes, and annotated genomes. VEHoP automates the generation of orthologous sequence alignments, concatenated matrices, and phylogenetic trees, streamlining phylogenomic analyses for researchers across disciplines. The tool aims to (1) expand taxonomic sampling by accommodating a wide range of input data types and (2) simplify phylogenomic workflows, making them accessible to researchers with varying levels of bioinformatic expertise. We evaluated VEHoPs performance using datasets from oysters, catfish, and insects, demonstrating its ability to produce robust phylogenetic trees with strong bootstrap support, outperforming assembly-free methods. Additionally, we applied VEHoP to reconstruct the phylogeny of the enigmatic deep-sea gastropod order Neomphalida, successfully resolving a well-supported phylogenetic backbone for this poorly understood group. VEHoP is freely available on GitHub (https://github.com/ylify/VEHoP), with dependencies easily installable via Bioconda.

bioinformatics↗

bioRxiv2024

Acceleration of genome rearrangement in clitellate annelids

Comparisons of multiple metazoan genomes have revealed the existence of ancestral linkage groups (ALGs), genomic scaffolds sharing sets of orthologous genes that have been inherited from ancestral animals for hundreds of millions of years (Simakov et al. 2022; Schultz et al. 2023) These ALGs have persisted across major animal taxa including Cnidaria, Deuterostomia, Ecdysozoa and Spiralia. Notwithstanding this general trend of chromosome-scale conservation, ALGs have been obliterated by extensive genome rearrangements in certain groups, most notably including Clitellata (oligochaetes and leeches), a group of easily overlooked invertebrates that is of tremendous ecological, agricultural and economic importance (Charles 2019; Barrett 2016). To further investigate these rearrangements, we have undertaken a comparison of 12 clitellate genomes (including four newly sequenced species) and 11 outgroup representatives. We show that these rearrangements began at the base of the Clitellata (rather than progressing gradually throughout polychaete annelids), that the inter-chromosomal rearrangements continue in several clitellate lineages and that these events have substantially shaped the evolution of the otherwise highly conserved Hox cluster.

evolutionary biology↗

bioRxiv2024

Multiple Displacement Amplification Facilitates SMRT Sequencing of Microscopic Animals and the Genome of the Gastrotrich Lepidodermella squamata (Dujardin, 1841)

BackgroundObtaining adequate DNA for long-read genome sequencing remains a roadblock to producing contiguous genomes from small-bodied organisms. Multiple displacement amplification (MDA) leverages Phi29 DNA polymerase to produce micrograms of DNA from picograms of input. Few genomes have been generated using this approach, due to concerns over biases in amplification related to GC and repeat content and chimera production. Here, we explored the utility of MDA for generating template DNA for PacBio HiFi sequencing using Caenorhabditis elegans (Nematoda) and Lepidodermella squamata (Gastrotricha). ResultsHiFi sequencing of libraries prepared from MDA DNA produced highly contiguous and complete genomes for both C. elegans (102 Mbp assembly; 336 contigs; N50 = 868 Kbp; L50 = 39; BUSCO_nematoda: S:92.2%, D:2.7%) and L. squamata (122 Mbp assembly; 157 contigs; N50 = 3.9 Mb; L50 = 13; BUSCO_metazoa: S: 78.0%, D: 2.8%). Amplified C. elegans reads mapped to the reference genome with a rate of 99.92% and coverage of 99.75% with just one read (of 708,811) inferred to be chimeric. Coverage uniformity was nearly identical for reads from MDA DNA and reads from pooled worm DNA when mapped to the reference genome. The genome of Lepidodermella squamata, the first of its phylum, was leveraged to infer the phylogenetic position of Gastrotricha, which has long been debated, as the sister taxon of Platyhelminthes. ConclusionsThis methodology will help generate contiguous genomes of microscopic taxa whose body size precludes standard long-read sequencing. L. squamata is an emerging model in evolutionary developmental biology and this genome will facilitate further work on this species.

genomics↗

bioRxiv2021

Benchmarking Oxford Nanopore read assemblers for high-quality molluscan genomes

Choosing the optimum assembly approach is essential to achieving a high-quality genome assembly suitable for comparative and evolutionary genomic investigations. Significant recent progress in long-read sequencing technologies such as PacBio and Oxford Nanopore Technologies (ONT) also brought about a large variety of assemblers. Although these have been extensively tested on model species such as Homo sapiens and Drosophila melanogaster, such benchmarking has not been done in Mollusca which lacks widely adopted model species. Molluscan genomes are notoriously rich in repeats and are often highly heterozygous, making their assembly challenging. Here, we benchmarked 10 assemblers based on ONT raw reads from two published molluscan genomes of differing properties, the gastropod Chrysomallon squamiferum (356.6Mb, 1.59% heterozygosity) and the bivalve Mytilus coruscus (1593Mb, 1.94% heterozygosity). By optimising the assembly pipeline, we greatly improved both genomes from previously published versions. Our results suggested that 40-50X of ONT reads are sufficient for high-quality genomes, with Flye being the recommended assembler for compact and less heterozygous genomes exemplified by C. squamiferum, while NextDenovo excelled for more repetitive and heterozygous molluscan genomes exemplified by M. coruscus. A phylogenomic analysis utilising the two updated genomes with other 32 published high-quality lophotrochozoan genomes resulted in maximum support across all nodes, and we show that improved genome quality also leads to more complete matrices for phylogenomic inferences. Our benchmarking will ensure the efficiency in future assemblies for molluscs and perhaps also other marine phyla with few genomes available.

genomics↗

bioRxiv2019

Pluripotency and the origin of animal multicellularity

The most widely held, but rarely tested, hypothesis for the origin of animals is that they evolved from a unicellular ancestor with an apical cilium surrounded by a microvillar collar that structurally resembled present-day sponge choanocytes and choanoflagellates1-4. Here we test this traditional view of the origin of the animal kingdom by comparing the transcriptomes, fates and behaviours of the three primary sponge cell types - choanocytes, pluripotent mesenchymal archeocytes and epithelial pinacocytes - with choanoflagellates and other unicellular holozoans. Unexpectedly, we find the transcriptome of sponge choanocytes is the least similar to the transcriptomes of choanoflagellates and is significantly enriched in genes unique to either animals or to sponges alone. In contrast, pluripotent archeocytes upregulate genes controlling cell proliferation and gene expression, as in other metazoan stem cells and in the proliferating stages of two closely-related unicellular holozoans, including a colonial choanoflagellate. In the context of the body plan of the sponge, Amphimedon queenslandica, we show that choanocytes appear late in development and are the result of a transdifferentiation event. They exist in a metastable state and readily transdifferentiate into archeocytes, which can differentiate into a range of other cell types. These sponge cell type conversions are similar to the temporal cell state changes that occur in many unicellular holozoans5. Together, these analyses offer no support for the homology of sponge choanocytes and choanoflagellates, nor for the view that the first multicellular animals were simple balls of cells with limited capacity to differentiate. Instead, our results are consistent with the first animal cell being able to transition between multiple states in a manner similar to modern transdifferentiating and stem cells.

evolutionary biology↗