Development of a Genotyping‐in‐Thousands by Sequencing (GT‐Seq) Panel for Identifying Individuals and Estimating Relatedness Among Alaska Black Bears (Ursus americanus)

ABSTRACT The management and conservation of large mammals, such as black bears (Ursus americanus), have long been informed by genetic estimates of population size and individual dispersal. Amplicon sequencing methods, also known as ‘genotyping‐in‐thousands‐by sequencing’ (GT‐seq), now enable the eff...

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Main Authors: Eleni L. Petrou, Colette D. Brandt, Timothy J. Spivey, Kristen M. Gruenthal, Cherie M. McKeeman, Sean D. Farley, David Battle, Cory Stantorf, Andrew M. Ramey
Format: Article
Language:English
Published: Wiley 2025-04-01
Series:Ecology and Evolution
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Online Access:https://doi.org/10.1002/ece3.71273
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author Eleni L. Petrou
Colette D. Brandt
Timothy J. Spivey
Kristen M. Gruenthal
Cherie M. McKeeman
Sean D. Farley
David Battle
Cory Stantorf
Andrew M. Ramey
author_facet Eleni L. Petrou
Colette D. Brandt
Timothy J. Spivey
Kristen M. Gruenthal
Cherie M. McKeeman
Sean D. Farley
David Battle
Cory Stantorf
Andrew M. Ramey
author_sort Eleni L. Petrou
collection DOAJ
description ABSTRACT The management and conservation of large mammals, such as black bears (Ursus americanus), have long been informed by genetic estimates of population size and individual dispersal. Amplicon sequencing methods, also known as ‘genotyping‐in‐thousands‐by sequencing’ (GT‐seq), now enable the efficient and cost‐effective genotyping of hundreds of loci and individuals in the same sequencing run. Here, we develop a GT‐seq panel for individual identification and kinship inference in Alaska black bears. Using genomic data from restriction site‐associated DNA sequencing of hunter‐harvested bears from Southcentral Alaska (n = 85), we identified 170 microhaplotype and single nucleotide polymorphism (SNP) loci that were highly heterozygous in local populations. To enable sexing of individuals, we also included a previously published sex‐linked locus in the GT‐seq panel. We empirically validated the GT‐seq panel using samples collected at different spatial scales. These samples included tissues (n = 82) obtained from bears within a small geographic area in Anchorage, Alaska, which were likely to be relatives as well as the hunter‐harvested samples collected from geographically widespread locations throughout Southcentral Alaska. Empirical validation indicated high genotyping success and genotype reproducibility across replicate subsamples. Computer simulations demonstrated that the GT‐seq panel had ample statistical power for distinguishing distinct individuals and first‐order relatives (parent‐offspring and full‐sibling pairs) from unrelated individuals. As a final proof of concept, the panel was used to identify individual bears and close kin sampled from urban and wild habitats in Anchorage, Alaska. We anticipate that the GT‐seq panel will be a useful genomic resource for the monitoring and management of Alaska black bear populations.
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spelling doaj-art-e98e2817244d4101b68c29df04a53d612025-08-20T02:35:33ZengWileyEcology and Evolution2045-77582025-04-01154n/an/a10.1002/ece3.71273Development of a Genotyping‐in‐Thousands by Sequencing (GT‐Seq) Panel for Identifying Individuals and Estimating Relatedness Among Alaska Black Bears (Ursus americanus)Eleni L. Petrou0Colette D. Brandt1Timothy J. Spivey2Kristen M. Gruenthal3Cherie M. McKeeman4Sean D. Farley5David Battle6Cory Stantorf7Andrew M. Ramey8U.S. Geological Survey, Alaska Science Center 4210 University Drive Anchorage Alaska USAU.S. air Force, Joint Base Elmendorf‐Richardson JBER Alaska USAAlaska Department of Fish and Game Anchorage Alaska USAAlaska Department of Fish and Game Anchorage Alaska USAU.S. Geological Survey, Alaska Science Center 4210 University Drive Anchorage Alaska USAAlaska Department of Fish and Game Anchorage Alaska USAAlaska Department of Fish and Game Anchorage Alaska USAAlaska Department of Fish and Game Anchorage Alaska USAU.S. Geological Survey, Alaska Science Center 4210 University Drive Anchorage Alaska USAABSTRACT The management and conservation of large mammals, such as black bears (Ursus americanus), have long been informed by genetic estimates of population size and individual dispersal. Amplicon sequencing methods, also known as ‘genotyping‐in‐thousands‐by sequencing’ (GT‐seq), now enable the efficient and cost‐effective genotyping of hundreds of loci and individuals in the same sequencing run. Here, we develop a GT‐seq panel for individual identification and kinship inference in Alaska black bears. Using genomic data from restriction site‐associated DNA sequencing of hunter‐harvested bears from Southcentral Alaska (n = 85), we identified 170 microhaplotype and single nucleotide polymorphism (SNP) loci that were highly heterozygous in local populations. To enable sexing of individuals, we also included a previously published sex‐linked locus in the GT‐seq panel. We empirically validated the GT‐seq panel using samples collected at different spatial scales. These samples included tissues (n = 82) obtained from bears within a small geographic area in Anchorage, Alaska, which were likely to be relatives as well as the hunter‐harvested samples collected from geographically widespread locations throughout Southcentral Alaska. Empirical validation indicated high genotyping success and genotype reproducibility across replicate subsamples. Computer simulations demonstrated that the GT‐seq panel had ample statistical power for distinguishing distinct individuals and first‐order relatives (parent‐offspring and full‐sibling pairs) from unrelated individuals. As a final proof of concept, the panel was used to identify individual bears and close kin sampled from urban and wild habitats in Anchorage, Alaska. We anticipate that the GT‐seq panel will be a useful genomic resource for the monitoring and management of Alaska black bear populations.https://doi.org/10.1002/ece3.71273amplicon sequencingGT‐seqindividual identificationparentagepopulation structureUrsus americanus
spellingShingle Eleni L. Petrou
Colette D. Brandt
Timothy J. Spivey
Kristen M. Gruenthal
Cherie M. McKeeman
Sean D. Farley
David Battle
Cory Stantorf
Andrew M. Ramey
Development of a Genotyping‐in‐Thousands by Sequencing (GT‐Seq) Panel for Identifying Individuals and Estimating Relatedness Among Alaska Black Bears (Ursus americanus)
Ecology and Evolution
amplicon sequencing
GT‐seq
individual identification
parentage
population structure
Ursus americanus
title Development of a Genotyping‐in‐Thousands by Sequencing (GT‐Seq) Panel for Identifying Individuals and Estimating Relatedness Among Alaska Black Bears (Ursus americanus)
title_full Development of a Genotyping‐in‐Thousands by Sequencing (GT‐Seq) Panel for Identifying Individuals and Estimating Relatedness Among Alaska Black Bears (Ursus americanus)
title_fullStr Development of a Genotyping‐in‐Thousands by Sequencing (GT‐Seq) Panel for Identifying Individuals and Estimating Relatedness Among Alaska Black Bears (Ursus americanus)
title_full_unstemmed Development of a Genotyping‐in‐Thousands by Sequencing (GT‐Seq) Panel for Identifying Individuals and Estimating Relatedness Among Alaska Black Bears (Ursus americanus)
title_short Development of a Genotyping‐in‐Thousands by Sequencing (GT‐Seq) Panel for Identifying Individuals and Estimating Relatedness Among Alaska Black Bears (Ursus americanus)
title_sort development of a genotyping in thousands by sequencing gt seq panel for identifying individuals and estimating relatedness among alaska black bears ursus americanus
topic amplicon sequencing
GT‐seq
individual identification
parentage
population structure
Ursus americanus
url https://doi.org/10.1002/ece3.71273
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