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      Insights into the species evolution of Calanus copepods in the northern seas revealed by de novo transcriptome sequencing

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          Abstract

          Copepods of the zooplankton genus Calanus play a key role in marine ecosystems in the northern seas. Although being among the most studied organisms on Earth, due to their ecological importance, genomic resources for Calanus spp. remain scarce, mostly due to their large genome size (from 6 to 12 Gbps). As an alternative to whole‐genome sequencing in Calanus spp., we sequenced and de novo assembled transcriptomes of five Calanus species: Calanus glacialis, C. hyperboreus, C. marshallae, C. pacificus, and Chelgolandicus. Functional assignment of protein families based on clusters of orthologous genes (COG) and gene ontology (GO) annotations showed analogous patterns of protein functions across species. Phylogenetic analyses using maximum likelihood (ML) of 191 protein‐coding genes mined from RNA‐seq data fully resolved evolutionary relationships among seven Calanus species investigated (five species sequenced for this study and two species with published datasets), with gene and site concordance factors showing that 109 out of 191 protein‐coding genes support a separation between three groups: the Cfinmarchicus group (including Cfinmarchicus, C. glacialis, and C. marshallae), the Chelgolandicus group (including Chelgolandicus, C. sinicus, and Cpacificus) and the monophyletic C. hyperboreus group. The tree topology obtained in ML analyses was similar to a previously proposed phylogeny based on morphological criteria and cleared certain ambiguities from past studies on evolutionary relationships among Calanus species.

          Abstract

          This study contributes and improve the currently available transcriptomic resources for Calanus spp. and explore de novo transcriptome data as an alternative to whole genome sequencing to infer evolutionary relationships within the genus Calanus. Here, we sequenced, assembled, and annotated de novo transcriptomes of two species for the first time ( C. hyperboreus and C. marshallae), and three species with some but limited transcriptomic data available ( Cglacialis, Chelgolandicus, and C. pacificus).

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          Cutadapt removes adapter sequences from high-throughput sequencing reads

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            IQ-TREE: A Fast and Effective Stochastic Algorithm for Estimating Maximum-Likelihood Phylogenies

            Large phylogenomics data sets require fast tree inference methods, especially for maximum-likelihood (ML) phylogenies. Fast programs exist, but due to inherent heuristics to find optimal trees, it is not clear whether the best tree is found. Thus, there is need for additional approaches that employ different search strategies to find ML trees and that are at the same time as fast as currently available ML programs. We show that a combination of hill-climbing approaches and a stochastic perturbation method can be time-efficiently implemented. If we allow the same CPU time as RAxML and PhyML, then our software IQ-TREE found higher likelihoods between 62.2% and 87.1% of the studied alignments, thus efficiently exploring the tree-space. If we use the IQ-TREE stopping rule, RAxML and PhyML are faster in 75.7% and 47.1% of the DNA alignments and 42.2% and 100% of the protein alignments, respectively. However, the range of obtaining higher likelihoods with IQ-TREE improves to 73.3-97.1%.
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              Trinity: reconstructing a full-length transcriptome without a genome from RNA-Seq data

              Massively-parallel cDNA sequencing has opened the way to deep and efficient probing of transcriptomes. Current approaches for transcript reconstruction from such data often rely on aligning reads to a reference genome, and are thus unsuitable for samples with a partial or missing reference genome. Here, we present the Trinity methodology for de novo full-length transcriptome reconstruction, and evaluate it on samples from fission yeast, mouse, and whitefly – an insect whose genome has not yet been sequenced. Trinity fully reconstructs a large fraction of the transcripts present in the data, also reporting alternative splice isoforms and transcripts from recently duplicated genes. In all cases, Trinity performs better than other available de novo transcriptome assembly programs, and its sensitivity is comparable to methods relying on genome alignments. Our approach provides a unified and general solution for transcriptome reconstruction in any sample, especially in the complete absence of a reference genome.

                Author and article information

                Contributors
                apollo.m.lizano@nord.no
                Journal
                Ecol Evol
                Ecol Evol
                10.1002/(ISSN)2045-7758
                ECE3
                Ecology and Evolution
                John Wiley and Sons Inc. (Hoboken )
                2045-7758
                22 February 2022
                February 2022
                : 12
                : 2 ( doiID: 10.1002/ece3.v12.2 )
                : e8606
                Affiliations
                [ 1 ] Faculty of Biosciences and Aquaculture Nord University Bodø Norway
                [ 2 ] Department of Medical Biochemistry and Microbiology Uppsala University Uppsala Sweden
                Author notes
                [*] [* ] Correspondence

                Apollo Marco Lizano, Faculty of Biosciences and Aquaculture, Nord University, Bodø, Norway.

                Email: apollo.m.lizano@ 123456nord.no

                Author information
                https://orcid.org/0000-0002-8216-3078
                https://orcid.org/0000-0002-0205-7663
                https://orcid.org/0000-0001-6719-2332
                Article
                ECE38606
                10.1002/ece3.8606
                8861592
                35228861
                1b48e9c0-7111-41a5-a7ff-8dedcd0384a4
                © 2022 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 14 January 2022
                : 27 September 2021
                : 19 January 2022
                Page count
                Figures: 5, Tables: 2, Pages: 15, Words: 11288
                Funding
                Funded by: Nord universitet , doi 10.13039/100015995;
                Categories
                Ecological Genetics
                Research Article
                Research Articles
                Custom metadata
                2.0
                February 2022
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.1.1 mode:remove_FC converted:22.02.2022

                Evolutionary Biology
                calanus,concordance factor,de novo transcriptome,phylotranscriptomics,rna‐seq

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