Molecular Method Based on Hydrolysis Probe Assays to Identify the Sex Chromosomes of Iberian Desman (Galemys pyrenaicus) Using Non-Invasive Sampling.

ABSTRACT Desmans belong to the subfamily Desmaninae, which are members of the family Talpidae. Desmans and moles show limited sexual dimorphism, making unclear sex discrimination by phenotypic assessment. The Iberian desman (Galemys pyrenaicus) is an endangered species with a severe population decli...

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Autores: Palacios-Gonzalez, María Jesús, Díaz-Caballero, José A., Espinosa, Antonio, Zalba, Francisco Javier, García-Zapata, Juan Luis, Fernádez-García , José Luis, Ripa Lopez-Barrantes, Adriana|||/items/37da2d92-f130-48c4-873b-810acb409892
Tipo de recurso: artículo
Fecha de publicación:2025
País:España
Institución:Universidad Alfonso X el Sabio
Repositorio:Repositorio Institucional de la Universidad Alfonso X el Sabio
Idioma:inglés
OAI Identifier:oai:archive.uax.com:20.500.12080/54506
Acceso en línea:https://hdl.handle.net/20.500.12080/54506
Access Level:acceso abierto
Palabra clave:DBX/DBY
Galemys pyrenaicus
Muestras no invasivas
RT-qPCR
Identificación del sexo
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spelling Molecular Method Based on Hydrolysis Probe Assays to Identify the Sex Chromosomes of Iberian Desman (Galemys pyrenaicus) Using Non-Invasive Sampling.Palacios-Gonzalez, María JesúsDíaz-Caballero, José A.Espinosa, AntonioZalba, Francisco JavierGarcía-Zapata, Juan LuisFernádez-García , José LuisRipa Lopez-Barrantes, Adriana|||/items/37da2d92-f130-48c4-873b-810acb409892DBX/DBYGalemys pyrenaicusMuestras no invasivasRT-qPCRIdentificación del sexoABSTRACT Desmans belong to the subfamily Desmaninae, which are members of the family Talpidae. Desmans and moles show limited sexual dimorphism, making unclear sex discrimination by phenotypic assessment. The Iberian desman (Galemys pyrenaicus) is an endangered species with a severe population decline. Knowledge of sex and sex ratio is essential for conservation and management. Based on these arguments and although previous conventional PCR studies amplifying DBX/DBY genes were relatively successful in sexing the desman, high-resolution sex-specific PCR has been requested. All these reasons encouraged us to develop new species-specific RT-qPCR assays by TaqMan probes to determine the sex in desman, especially with genetic material from non invasive samples. Accordingly, efficiency, limit of detection (LOD), specificity, and DNA analysis from faeces were verified. The target genes DBX and DBY were amplified with gDNA from both sexes, with Y-chromosome consistently absent in the female. Despite the modest efficiency, regression analysis (R2 > 0.999) indicated a linear range of the DBX and DBY assays extending from 20 to 0.2 ng/µL DNA. LOD analyses estimated that twice as much gDNA was needed in males as in females for DBX detection. Paradoxically, the Y-chromosome required three times as much gDNA as the X-chromosome using a male sample. Therefore, an unexpected dosage imbalance in the genome in favour of the X chromosome was discussed in light of an apparent multicopy nature of the DBX gene and with a sexing success rate of 49.9% of the non-invasive samples, supporting Fisher’s principle for the mammalian XX/XY sex system, as expected.WileyUAX -Campus MadridUAX Archive20252025-01-0120252025-01-01journal articlehttp://purl.org/coar/resource_type/c_6501info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.12080/54506reponame:Repositorio Institucional de la Universidad Alfonso X el Sabioinstname:Universidad Alfonso X el SabioInglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:archive.uax.com:20.500.12080/545062026-05-28T12:40:21Z
dc.title.none.fl_str_mv Molecular Method Based on Hydrolysis Probe Assays to Identify the Sex Chromosomes of Iberian Desman (Galemys pyrenaicus) Using Non-Invasive Sampling.
title Molecular Method Based on Hydrolysis Probe Assays to Identify the Sex Chromosomes of Iberian Desman (Galemys pyrenaicus) Using Non-Invasive Sampling.
spellingShingle Molecular Method Based on Hydrolysis Probe Assays to Identify the Sex Chromosomes of Iberian Desman (Galemys pyrenaicus) Using Non-Invasive Sampling.
Palacios-Gonzalez, María Jesús
DBX/DBY
Galemys pyrenaicus
Muestras no invasivas
RT-qPCR
Identificación del sexo
title_short Molecular Method Based on Hydrolysis Probe Assays to Identify the Sex Chromosomes of Iberian Desman (Galemys pyrenaicus) Using Non-Invasive Sampling.
title_full Molecular Method Based on Hydrolysis Probe Assays to Identify the Sex Chromosomes of Iberian Desman (Galemys pyrenaicus) Using Non-Invasive Sampling.
title_fullStr Molecular Method Based on Hydrolysis Probe Assays to Identify the Sex Chromosomes of Iberian Desman (Galemys pyrenaicus) Using Non-Invasive Sampling.
title_full_unstemmed Molecular Method Based on Hydrolysis Probe Assays to Identify the Sex Chromosomes of Iberian Desman (Galemys pyrenaicus) Using Non-Invasive Sampling.
title_sort Molecular Method Based on Hydrolysis Probe Assays to Identify the Sex Chromosomes of Iberian Desman (Galemys pyrenaicus) Using Non-Invasive Sampling.
dc.creator.none.fl_str_mv Palacios-Gonzalez, María Jesús
Díaz-Caballero, José A.
Espinosa, Antonio
Zalba, Francisco Javier
García-Zapata, Juan Luis
Fernádez-García , José Luis
Ripa Lopez-Barrantes, Adriana|||/items/37da2d92-f130-48c4-873b-810acb409892
author Palacios-Gonzalez, María Jesús
author_facet Palacios-Gonzalez, María Jesús
Díaz-Caballero, José A.
Espinosa, Antonio
Zalba, Francisco Javier
García-Zapata, Juan Luis
Fernádez-García , José Luis
Ripa Lopez-Barrantes, Adriana|||/items/37da2d92-f130-48c4-873b-810acb409892
author_role author
author2 Díaz-Caballero, José A.
Espinosa, Antonio
Zalba, Francisco Javier
García-Zapata, Juan Luis
Fernádez-García , José Luis
Ripa Lopez-Barrantes, Adriana|||/items/37da2d92-f130-48c4-873b-810acb409892
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv UAX -Campus Madrid
UAX Archive
dc.subject.none.fl_str_mv DBX/DBY
Galemys pyrenaicus
Muestras no invasivas
RT-qPCR
Identificación del sexo
topic DBX/DBY
Galemys pyrenaicus
Muestras no invasivas
RT-qPCR
Identificación del sexo
description ABSTRACT Desmans belong to the subfamily Desmaninae, which are members of the family Talpidae. Desmans and moles show limited sexual dimorphism, making unclear sex discrimination by phenotypic assessment. The Iberian desman (Galemys pyrenaicus) is an endangered species with a severe population decline. Knowledge of sex and sex ratio is essential for conservation and management. Based on these arguments and although previous conventional PCR studies amplifying DBX/DBY genes were relatively successful in sexing the desman, high-resolution sex-specific PCR has been requested. All these reasons encouraged us to develop new species-specific RT-qPCR assays by TaqMan probes to determine the sex in desman, especially with genetic material from non invasive samples. Accordingly, efficiency, limit of detection (LOD), specificity, and DNA analysis from faeces were verified. The target genes DBX and DBY were amplified with gDNA from both sexes, with Y-chromosome consistently absent in the female. Despite the modest efficiency, regression analysis (R2 > 0.999) indicated a linear range of the DBX and DBY assays extending from 20 to 0.2 ng/µL DNA. LOD analyses estimated that twice as much gDNA was needed in males as in females for DBX detection. Paradoxically, the Y-chromosome required three times as much gDNA as the X-chromosome using a male sample. Therefore, an unexpected dosage imbalance in the genome in favour of the X chromosome was discussed in light of an apparent multicopy nature of the DBX gene and with a sexing success rate of 49.9% of the non-invasive samples, supporting Fisher’s principle for the mammalian XX/XY sex system, as expected.
publishDate 2025
dc.date.none.fl_str_mv 2025
2025-01-01
2025
2025-01-01
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/20.500.12080/54506
url https://hdl.handle.net/20.500.12080/54506
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv reponame:Repositorio Institucional de la Universidad Alfonso X el Sabio
instname:Universidad Alfonso X el Sabio
instname_str Universidad Alfonso X el Sabio
reponame_str Repositorio Institucional de la Universidad Alfonso X el Sabio
collection Repositorio Institucional de la Universidad Alfonso X el Sabio
repository.name.fl_str_mv
repository.mail.fl_str_mv
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