Discriminating larvae of two syntopic  Cychramus  species (Coleoptera, Nitidulidae) by means of bar-HRM analysis

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Abstract

Molecular genetic methods are increasingly used to supplement or substitute classical morphology-based species identification. Here, we employ a COI mini-barcode coupled high-resolution melting analysis to quickly, cost-efficiently and reliably determine larvae of two closely related Cychramus (Coleoptera, Nitidulidae) species. Euclidean distance comparison (p < 0.01) and a Welch t-test of the melting point temperatures (p < 0.01) provide highly significant statistical evidence for species specific differences in melting and fluorescence curves, thus allowing the assignment of larvae to either of the two species. This protocol serves as a fast, low-cost and low-tech method to discriminate between pairs or groups of closely related species and can be adapted and applied to various ecological research questions.

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MEGA6: Molecular Evolutionary Genetics Analysis version 6.0.

Koichiro Tamura, Glen Stecher, Daniel Walter Peterson … (2013)

We announce the release of an advanced version of the Molecular Evolutionary Genetics Analysis (MEGA) software, which currently contains facilities for building sequence alignments, inferring phylogenetic histories, and conducting molecular evolutionary analysis. In version 6.0, MEGA now enables the inference of timetrees, as it implements the RelTime method for estimating divergence times for all branching points in a phylogeny. A new Timetree Wizard in MEGA6 facilitates this timetree inference by providing a graphical user interface (GUI) to specify the phylogeny and calibration constraints step-by-step. This version also contains enhanced algorithms to search for the optimal trees under evolutionary criteria and implements a more advanced memory management that can double the size of sequence data sets to which MEGA can be applied. Both GUI and command-line versions of MEGA6 can be downloaded from www.megasoftware.net free of charge.

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Hadley Wickham, Mara Averick, Jennifer Bryan … (2019)

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Biological identifications through DNA barcodes.

Paul Hebert, Alina Cywinska, Shelley Ball … (2003)

Although much biological research depends upon species diagnoses, taxonomic expertise is collapsing. We are convinced that the sole prospect for a sustainable identification capability lies in the construction of systems that employ DNA sequences as taxon 'barcodes'. We establish that the mitochondrial gene cytochrome c oxidase I (COI) can serve as the core of a global bioidentification system for animals. First, we demonstrate that COI profiles, derived from the low-density sampling of higher taxonomic categories, ordinarily assign newly analysed taxa to the appropriate phylum or order. Second, we demonstrate that species-level assignments can be obtained by creating comprehensive COI profiles. A model COI profile, based upon the analysis of a single individual from each of 200 closely allied species of lepidopterans, was 100% successful in correctly identifying subsequent specimens. When fully developed, a COI identification system will provide a reliable, cost-effective and accessible solution to the current problem of species identification. Its assembly will also generate important new insights into the diversification of life and the rules of molecular evolution.

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Author and article information

Contributors

Lukas Zangl:

ORCID: https://orcid.org/0000-0002-1175-564X

lukas.zangl@uni-graz.at

Stephan Koblmüller:

ORCID: https://orcid.org/0000-0002-1024-3220

stephan.koblmueller@uni-graz.at

Journal

Journal ID (nlm-ta): Mol Biol Rep

Journal ID (iso-abbrev): Mol Biol Rep

Title: Molecular Biology Reports

Publisher: Springer Netherlands (Dordrecht )

ISSN (Print): 0301-4851

ISSN (Electronic): 1573-4978

Publication date (Electronic): 8 September 2020

Publication date PMC-release: 8 September 2020

Publication date (Print): 2020

Volume: 47

Issue: 10

Pages: 8251-8257

Affiliations

[1 ]GRID grid.5110.5, ISNI 0000000121539003, Institute of Biology, , University of Graz, ; Universitätsplatz 2, 8010 Graz, Austria

[2 ]GRID grid.472881.0, ISNI 0000 0001 1348 1753, Universalmuseum Joanneum, Studienzentrum Naturkunde, ; Weinzöttlstraße 16, 8045 Graz, Austria

[3 ]ÖKOTEAM - Institute for Animal Ecology and Landscape Planning, Bergmanngasse 22, 8010 Graz, Austria

[4 ]Present Address: 4651 Stadl-Paura, Austria

[5 ]GRID grid.5110.5, ISNI 0000000121539003, Institute of Biology, , University of Graz, ; Schubertstraße 51, 8010 Graz, Austria

Author information

Lukas Zangl https://orcid.org/0000-0002-1175-564X

Hannes Oberreiter https://orcid.org/0000-0003-1019-1245

Edith Stabentheiner https://orcid.org/0000-0001-5228-8390

Christian Sturmbauer https://orcid.org/0000-0003-4170-2765

Stephan Koblmüller https://orcid.org/0000-0002-1024-3220

Article

Publisher ID: 5786

DOI: 10.1007/s11033-020-05786-9

PMC ID: 7588358

PubMed ID: 32901359

SO-VID: 18192e31-6aef-45cf-835b-8ce11d1c9887

License:

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

History

Date received : 16 June 2020

Date accepted : 28 August 2020

Funding

Funded by: Austrian Federal Ministry of Science, Research and Economy

Custom metadata

ScienceOpen disciplines: Molecular biology

Keywords: coleoptera,dna-barcoding,high-resolution melting analysis,larvae,mini-barcodes,sap beetles

Data availability:

ScienceOpen disciplines: Molecular biology

Keywords: coleoptera, dna-barcoding, high-resolution melting analysis, larvae, mini-barcodes, sap beetles

Discriminating larvae of two syntopic Cychramus species (Coleoptera, Nitidulidae) by means of bar-HRM analysis

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Abstract

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Coleoptera

Most cited references 24

MEGA6: Molecular Evolutionary Genetics Analysis version 6.0.

Welcome to the Tidyverse

Biological identifications through DNA barcodes.

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