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      18S-NemaBase: Curated 18S rRNA Database of Nematode Sequences

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          Abstract

          Nematodes are the most abundant and diverse animals on the planet but lack representation in biodiversity research. This presents a problem for studying nematode diversity, particularly when molecular tools (i.e., barcoding and metabarcoding) rely on well-populated and curated reference databases, which are absent for nematodes. To improve molecular identification and the assessment of nematode diversity, we created and curated an 18S rRNA database specific to nematodes (18S-NemaBase) using sequences sourced from the most recent publicly available 18S rRNA SILVA v138 database. As part of the curation process, taxonomic strings were standardized to contain a fixed number of taxonomic ranks relevant to nematology and updated for the most recent accepted nematode classifications. In addition, apparent erroneous sequences were removed. To test the efficacy and accuracy of 18S-NemaBase, we compared it to an older but also curated SILVA v111 and the newest SILVA v138 by assigning taxonomies and analyzing the diversity of a nematode dataset from the Western Nebraska Sandhills. We showed that 18S-NemaBase provided more accurate taxonomic assignments and diversity assessments than either version of SILVA, with a much easier workflow and no need for manual corrections. Additionally, observed diversity further improved when 18S-NemaBase was supplemented with reference sequences from nematodes present in the study site. Although the 18S-NemaBase is a step in the right direction, a concerted effort to increase the number of high-quality, accessible, full-length nematode reference sequences is more important now than ever.

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          MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability

          We report a major update of the MAFFT multiple sequence alignment program. This version has several new features, including options for adding unaligned sequences into an existing alignment, adjustment of direction in nucleotide alignment, constrained alignment and parallel processing, which were implemented after the previous major update. This report shows actual examples to explain how these features work, alone and in combination. Some examples incorrectly aligned by MAFFT are also shown to clarify its limitations. We discuss how to avoid misalignments, and our ongoing efforts to overcome such limitations.
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            DADA2: High resolution sample inference from Illumina amplicon data

            We present DADA2, a software package that models and corrects Illumina-sequenced amplicon errors. DADA2 infers sample sequences exactly, without coarse-graining into OTUs, and resolves differences of as little as one nucleotide. In several mock communities DADA2 identified more real variants and output fewer spurious sequences than other methods. We applied DADA2 to vaginal samples from a cohort of pregnant women, revealing a diversity of previously undetected Lactobacillus crispatus variants.
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              Cutadapt removes adapter sequences from high-throughput sequencing reads

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                Author and article information

                Contributors
                Journal
                J Nematol
                J Nematol
                jofnem
                jofnem
                Journal of Nematology
                Sciendo
                0022-300X
                2640-396X
                21 April 2023
                February 2023
                : 55
                : 1
                : 20230006
                Affiliations
                Department of Entomology and Nematology, University of Florida , FL 32611 United States of America
                Department of Plant Pathology, University of Nebraska-Lincoln , NE 68588 United States of America
                Plant Sciences Unit, Flanders Research Institute for Agriculture, Fisheries and Food (ILVO) , Burg. Van Gansberghelaan 96, 9820 Merelbeke, Belgium
                Author notes

                Support Received from the University of Florida, University of Nebraska Collaborative Initiative Seed Grant Program, and the European H2020 program.

                This paper was edited by Erik C Andersen.

                Article
                jofnem-2023-0006
                10.2478/jofnem-2023-0006
                10152464
                37143483
                cc93a63d-498e-445c-9531-76c5ed253207
                © 2023 Gattoni et al., published by Sciendo

                This work is licensed under the Creative Commons Attribution 4.0 International License.

                History
                : 1 August 2022
                Page count
                Pages: 16
                Categories
                Research Paper

                biodiversity,database,ecology,metabarcoding,nematodes
                biodiversity, database, ecology, metabarcoding, nematodes

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