Tracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.

Eupsophus is a genus of ground frogs, endemic from Southern Chile and Argentina. The taxonomy and systematic of the genus have been controversial since its founding. Currently, Eupsophus is composed by 10 species, which have been match up into two groups: The roseus group that comprise eight species...

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Autor: Quercia-Raty, Camila
Tipo de recurso: tesis de maestría
Estado:Versión publicada
Fecha de publicación:2019
País:Chile
OAI Identifier:oai:repositorio.anid.cl:10533/246639
Acceso en línea:https://hdl.handle.net/10533/246639
Access Level:acceso abierto
Palabra clave:Ciencias Naturales
Otras Ciencias Naturales
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dc.title.es_CL.fl_str_mv Tracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.
title Tracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.
spellingShingle Tracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.
Quercia-Raty, Camila
Ciencias Naturales
Otras Ciencias Naturales
title_short Tracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.
title_full Tracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.
title_fullStr Tracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.
title_full_unstemmed Tracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.
title_sort Tracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.
dc.creator.none.fl_str_mv Quercia-Raty, Camila
author Quercia-Raty, Camila
author_facet Quercia-Raty, Camila
author_role author
dc.contributor.advisor.none.fl_str_mv Nuñez, José J.
dc.contributor.institution.es_CL.fl_str_mv UNIVERSIDAD AUSTRAL DE CHILE
dc.subject.oecd1n.es_CL.fl_str_mv Ciencias Naturales
topic Ciencias Naturales
Otras Ciencias Naturales
dc.subject.oecd2n.es_CL.fl_str_mv Otras Ciencias Naturales
description Eupsophus is a genus of ground frogs, endemic from Southern Chile and Argentina. The taxonomy and systematic of the genus have been controversial since its founding. Currently, Eupsophus is composed by 10 species, which have been match up into two groups: The roseus group that comprise eight species (2n=30), and the vertebralis group with two species E. vertebralis, and E. emiliopugini (2n=28). Within the vertebralis group, geographic distribution, morphologic and genetic variation remain unclear and without hypotheses about the relationships between these lineages. Moreover, morphology karyotype description made by Giemsa stained, evidenced differences on the fundamental number (FN) between E. vertebralis (FN= 54) and E. emiliopugini (FN= 56). Nevertheless, the consistency of this report at intraspecific levels and with others cytogenetics techniques remain unexplored. Thus, the goals of this thesis were, on the one hand, to evaluated phylogenetic relationships, diversification times, and to determine species limits of Eupsophus vertebralis and E. emiliopugini. On the other hand, it was proposed to determine the presences or absences of specie specific cytogenetics patterns for the vertebralis group, and test hypothesis about chromosomal rearrangements, considering individuals from different localities and sexes. For the molecular analyses, three mitochondrial (D-loop, cytb, and coI) and two nuclear (crybA1, and pomc) markers, from 91 individuals of the vertebralis group collected in 19 localities, were sequenced. Then, it was performed phylogenetics analyses with Maximum Likelihood (ML) and Bayesian Inference (IB), and species delimitation analyses using unilocus and multilocus coalescent approaches. For its part, for the cytogenetic analyses it was employed conventional (Giemsa staining, C-banding and, Ag-NOR) and cytomolecular (fluorescence in situ hybridization FISH, using telomeric and 28S ribosomal probes) techniques, on metaphasic plates of 23 individuals of the vertebralis group from 15 different localities. ML and IB phylogenetic reconstructions recovered the monophyly of the vertebralis group (Bootstrap: 100%; Bayesian posterior Probability PP: 1.0), but not the reciprocal monophyly of its species (Bootstrap: < 50; PP: < 80), exhibiting a polyphyletic pattern within the vertebralis group. Incongruence among the six species delimitation analyses carried out, detecting from one to eleven species (Max. congruence index= 0.59, for one putative species; Min. congruence index= 0.18, for eleven putative species). Also, the divergence time between E. emiliopugini and E. vertebralis (0.040 Mya) indicates an evolutionary history probably molded by the interglacial periods of Late Pleistocene. On the other hand, FN differences between E. vertebralis and E. emiliopugini were corroborated through Giemsa staining and C-banding (FN= 54 and 56, respectively). FISH with 28S rDNA probes confirmed the active NORs signal for E. vertebralis and E. emiliopugini (in accordance with the Ag-NORs staining results). Nevertheless, we found polymorphisms relative to the numbers and location of 28S rDNA (four signals) and active NORs (two signals) in one specimen of E. emiliopugini from Puyehue locality. This intraspecific chromosomal variation could be explained by chromosomal rearrangements or derived from hybridization process. FISH using telomeric probe over spreads from both species detected no interstitial fluorescent signals, but clearly stained telomeric regions. The molecular results of this study, suggest low genetic differentiation and the early diversification for the vertebralis group. Also, the cytogenetics results corroborate that the difference in FN between the karyotype of E. vertebralis and E. emiliopugini is conserved at interspecific level. Nonetheless, intraspecific differences for the NORs position were found in one sample of E. emliopugini from Puyehue locality. Therefore, the results do not provide sufficient evidence to support E. emiliopugini and E. vertebralis as one or two species. Hence, the speciation process of E. vertebralis and E. emiliopugini lineages are discussed under an integrative framework, considering the phylogenetic relationships, evolutionary history, geographic antecedents, cytogenetic, and ethologic (calls) data. Finally, new concerning lineage for conservation efforts are proposed.
publishDate 2019
dc.date.issued.es_CL.fl_str_mv 2019
dc.date.accessioned.none.fl_str_mv 2021-04-05T21:50:17Z
2022-08-16T18:21:30Z
dc.date.available.none.fl_str_mv 2021-04-05T21:50:17Z
2022-08-16T18:21:30Z
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dc.type.tesis.none.fl_str_mv Tesis
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dc.identifier.folio.es_CL.fl_str_mv 22180766
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/10533/246639
identifier_str_mv 22180766
url https://hdl.handle.net/10533/246639
dc.relation.none.fl_str_mv instname: Conicyt
reponame: Repositorio Digital RI2.0
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spelling UNIVERSIDAD AUSTRAL DE CHILEQuercia-Raty, Camila2019https://hdl.handle.net/10533/246639http://purl.org/coar/access_right/c_abf2Otras Ciencias NaturalesCiencias NaturalesTracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.Nuñez, José J.UNIVERSIDAD AUSTRAL DE CHILEChileQuercia-Raty, Camila2021-04-05T21:50:17Z2022-08-16T18:21:30Z2021-04-05T21:50:17Z2022-08-16T18:21:30Z2019Eupsophus is a genus of ground frogs, endemic from Southern Chile and Argentina. The taxonomy and systematic of the genus have been controversial since its founding. Currently, Eupsophus is composed by 10 species, which have been match up into two groups: The roseus group that comprise eight species (2n=30), and the vertebralis group with two species E. vertebralis, and E. emiliopugini (2n=28). Within the vertebralis group, geographic distribution, morphologic and genetic variation remain unclear and without hypotheses about the relationships between these lineages. Moreover, morphology karyotype description made by Giemsa stained, evidenced differences on the fundamental number (FN) between E. vertebralis (FN= 54) and E. emiliopugini (FN= 56). Nevertheless, the consistency of this report at intraspecific levels and with others cytogenetics techniques remain unexplored. Thus, the goals of this thesis were, on the one hand, to evaluated phylogenetic relationships, diversification times, and to determine species limits of Eupsophus vertebralis and E. emiliopugini. On the other hand, it was proposed to determine the presences or absences of specie specific cytogenetics patterns for the vertebralis group, and test hypothesis about chromosomal rearrangements, considering individuals from different localities and sexes. For the molecular analyses, three mitochondrial (D-loop, cytb, and coI) and two nuclear (crybA1, and pomc) markers, from 91 individuals of the vertebralis group collected in 19 localities, were sequenced. Then, it was performed phylogenetics analyses with Maximum Likelihood (ML) and Bayesian Inference (IB), and species delimitation analyses using unilocus and multilocus coalescent approaches. For its part, for the cytogenetic analyses it was employed conventional (Giemsa staining, C-banding and, Ag-NOR) and cytomolecular (fluorescence in situ hybridization FISH, using telomeric and 28S ribosomal probes) techniques, on metaphasic plates of 23 individuals of the vertebralis group from 15 different localities. ML and IB phylogenetic reconstructions recovered the monophyly of the vertebralis group (Bootstrap: 100%; Bayesian posterior Probability PP: 1.0), but not the reciprocal monophyly of its species (Bootstrap: < 50; PP: < 80), exhibiting a polyphyletic pattern within the vertebralis group. Incongruence among the six species delimitation analyses carried out, detecting from one to eleven species (Max. congruence index= 0.59, for one putative species; Min. congruence index= 0.18, for eleven putative species). Also, the divergence time between E. emiliopugini and E. vertebralis (0.040 Mya) indicates an evolutionary history probably molded by the interglacial periods of Late Pleistocene. On the other hand, FN differences between E. vertebralis and E. emiliopugini were corroborated through Giemsa staining and C-banding (FN= 54 and 56, respectively). FISH with 28S rDNA probes confirmed the active NORs signal for E. vertebralis and E. emiliopugini (in accordance with the Ag-NORs staining results). Nevertheless, we found polymorphisms relative to the numbers and location of 28S rDNA (four signals) and active NORs (two signals) in one specimen of E. emiliopugini from Puyehue locality. This intraspecific chromosomal variation could be explained by chromosomal rearrangements or derived from hybridization process. FISH using telomeric probe over spreads from both species detected no interstitial fluorescent signals, but clearly stained telomeric regions. The molecular results of this study, suggest low genetic differentiation and the early diversification for the vertebralis group. Also, the cytogenetics results corroborate that the difference in FN between the karyotype of E. vertebralis and E. emiliopugini is conserved at interspecific level. Nonetheless, intraspecific differences for the NORs position were found in one sample of E. emliopugini from Puyehue locality. Therefore, the results do not provide sufficient evidence to support E. emiliopugini and E. vertebralis as one or two species. Hence, the speciation process of E. vertebralis and E. emiliopugini lineages are discussed under an integrative framework, considering the phylogenetic relationships, evolutionary history, geographic antecedents, cytogenetic, and ethologic (calls) data. Finally, new concerning lineage for conservation efforts are proposed.Eupsophus es un género de ranas de hojarasca, endémicas del Sur de Chile y Argentina. La taxonomía y sistemática de este grupo ha sido controversial desde sus inicios. Actualmente son 10 las especies reconocidas para este género, las cuales clásicamente son divididas en dos grupos: El grupo roseus que comprende ocho especies (2n=30) y el grupo vertebralis compuesto por dos especies, E. vertebralis y E. emiliopugini (2n=28). Dentro del grupo vetebralis, la distribución geográfica, las características morfológicas y las variaciones genéticas permanecen confusas y sin hipótesis filogenéticas acerca de las relaciones entre sus linajes. Además, la comparación de los cariotipos entre ambas especies, utilizando citogenética clásica (tinción Giemsa), evidenció diferencias en el número fundamental de cromosomas (FN) entre los cariotipos de E. vertebralis (FN= 54) y E. emiliopugini (FN= 56). Sin embargo, la consistencia de estos reportes aún no ha sido explorada a nivel intraespecífico, así como tampoco mediante otras técnicas de citogenética molecular. Los objetivos abordados en este trabajo de tesis fueron, por una parte, evaluar las relaciones filogenéticas, los tiempos de divergencia, y determinar los límites de especies para E. vertebralis y E. emiliopugini. Por otra parte, se propuso determinar la presencia o ausencia de patrones citogenéticos especie específicos para el grupo vertebralis y probar posibles hipótesis acerca de re-arreglos cromosómicos, considerando individuos de diferentes poblaciones y sexos. Para los análisis moleculares, se secuenciaron tres marcadores mitocondriales (D-loop, cytb y coI) y dos nucleares (CrybA1 y pomc) para 91 individuos del grupo vertebralis, colectados de 19 localidades diferentes. Luego, se realizaron análisis filogenéticos de Máxima Verosimilitud (ML) e Inferencia Bayesiana (IB) y análisis de delimitación de especies unilocus y multilocus coalescente. Por otro lado, para los análisis citogenéticos, se emplearon técnicas clásicas (Tinción Giemsa, Bandeo C y Ag-NORs) y moleculares (hibridación in situ fluorescente FISH, usando zonas teloméricas y ribosomales) sobre placas metafásicas de 23 individuos del grupo vertebralis proveniente de 15 localidades diferentes. Las reconstrucciones filogenéticas de ML e IB, recuperaron la monofilia del grupo vertebralis (Bootstrap: 100%; Probabilidad a posterior Bayesiana PP: 1.0), no así de sus especies (Bootstrap: < 50; PP: < 80) exhibiendo un patrón polifilético dentro del grupo. Los resultados de los seis análisis de delimitación de especies fueron incongruentes entre ellos, recuperando desde una a 11 especies putativas (Máximo índice de congruencia para una especie= 0.59; Mínimo índice de congruencia para once especies= 0.18). El tiempo de divergencia entre E. emiliopugini y E. vertebralis (0.040 Mya) indicaría que la historia evolutiva de este grupo fue moldeada por periodos interglaciares del Pleistoceno tardío. Por otro lado, las diferencias en FN entre los cariotipos de ambas especies, fueron corroborados mediante la tinción de Giemnsa y Bandeos C (E. vertebralis FN= 54; E. emiliopugini FN=56). Los resultados de FISH, usando la sonda ribosomal 28S rDNA, confirmaron la señal obtenida para los NOR activos (de acuerdo con la tinción Ag-NORs) en E. vertebralis y en E. emiliopugini. Aún más, estos resultados mostraron un polimorfismo en relación al número y a la posición de los genes 28S rDNA (cuatro señales) y de los NORs (dos señales) activos para un espécimen de E. emiliopugini, colectado en la localidad de Puyehue. Esta variación cromosómica a nivel intraespecífico, podría explicarse por la ocurrencia de arreglos cromosómicos o procesos de hibridación. De los resultados de FISH con sonda telomérica, no se encontraron señales fluorescentes intersticiales, pero si teloméricas terminales para ambas especies. En definitiva, los resultados genético moleculares sugieren baja diferenciación genética y diversificación reciente para el grupo E. vertebralis. Además, los resultados citogenéticos corroboraron que la diferencia en el FN de los cariotipos de E. emiliopugini y E. vertebralis, es conservada a nivel intraespecífico, respectivamente. Sin embargo, diferencias intraespecíficas para la posición de NORs en E. emiliopugini fueron encontradas (solo una muestra analizada fue polimórfica). Por lo tanto, los resultados encontrados no brindan evidencia suficiente que soporte a E. emiliopugini y E. vertebralis como una o como dos especies. Por consiguiente, el proceso de especiación de los linajes de E. vertebralis and E. emiliopugini es discutido bajo un marco integrativo, considerando relaciones filogenéticas, historia evolutiva, antecedentes geográficos, citogenéticos y etológicos (cantos). Finalmente son mencionados nuevos linajes con importancia para conservación.22180766https://hdl.handle.net/10533/246639instname: Conicytreponame: Repositorio Digital RI2.0info:eu-repo/grantAgreement//22180766info:eu-repo/semantics/dataset/hdl.handle.net/10533/93488info:eu-repo/semantics/openAccessAttribution-NonCommercial-NoDerivs 3.0 Chilehttp://creativecommons.org/licenses/by-nc-nd/3.0/cl/Ciencias NaturalesOtras Ciencias NaturalesTracing the evolutionary history of two endemic ground frogs of temperate forest of southern Chile, through molecular and cytogenetic approaches.info:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/publishedVersionTesisTesishttps://hdl.handle.net/10533/24663911424ce9-522e-4422-9ccc-84c6fb39d67cvirtual::57721-111424ce9-522e-4422-9ccc-84c6fb39d67cvirtual::57721-1THUMBNAILTesis_QuerciaRatyCA.pdf.jpgIM Thumbnailimage/jpeg3781https://repositorio.anid.cl/bitstreams/4d1f6119-74df-442b-b442-e98aa8692c10/downloada17be843890e2d0317ed7dfdb253d1a8MD51TEXTTesis_QuerciaRatyCA.pdf.txtExtracted texttext/plain203680https://repositorio.anid.cl/bitstreams/4dbce0f9-fb6d-4ad8-a405-672052f077f5/download693a922335d10d28a0aae09b405d3609MD52CC-LICENSElicense_rdfapplication/octet-stream1232https://repositorio.anid.cl/bitstreams/a9cfeee6-65aa-44c1-871e-9bbc4b94de3d/downloadf97bcfdf58f3e17b5cec231112dab5b1MD53LICENSElicense.txttext/plain1779https://repositorio.anid.cl/bitstreams/40d049de-dede-4011-a75d-84d3d9563649/download593a6e7305c66c56041a9f9e15a649c1MD54ORIGINALTesis_QuerciaRatyCA.pdfTesis Formato Magisterapplication/pdf10511019https://repositorio.anid.cl/bitstreams/d0fd5d88-99d7-42b0-aba9-1b4385ae11e4/download1717882710fe784669dd10ea15e64d48MD5510533/246639oai:repositorio.anid.cl:10533/2466392023-07-24 10:51:07.923http://creativecommons.org/licenses/by-nc-nd/3.0/cl/info:eu-repo/semantics/openAccesshttps://repositorio.anid.clRepositorio ANIDaletelier@anid.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