José J. Nuñez

Amphibian phylogeography in the Antipodes: Refugia and postglacial colonization explain mitochondrial haplotype distribution in the Patagonian frog Eupsophus calcaratus (Cycloramphidae)

Climatic oscillations, heterogeneity in elevation, topographical position, and isolation time in southwestern Patagonia have been important in promoting diversification of the biota. Geological studies have shown that this region had wide ice-free areas during periods of the last glacial maximum and provided forested refugia for the biota during Pleistocene glaciations. In this study, we sampled the endemic frog Eupsophus calcaratus from 20 localities, covering most of its distribution and including glaciated and non-glaciated regions. We collected DNA sequences for three mitochondrial regions (D-loop, cyt b, 16S), and describe patterns of variation consistent with a history of both the displacement to glacial refugia and recent recolonization to extensively glaciated regions. The inferred demographic history and divergence times of the lineages of E. calcaratus suggest that the Pleistocene had profound effects on the genetic patterns within this taxon in which some populations were able to survive in refugia within colder regions followed by demographic increases but without evidence of significant range expansion. The mtDNA gene tree recovers six major haploclades of E. calcaratus, which we consider diagnostic of species lineages. These results contribute to our understanding of how geological events, predominately glacial oscillations, have influenced current population structure of a broad-ranging, ectothermic vertebrate in the Valdivian Forest region of southern South America.

Phylogenetic relationships of a Patagonian frog radiation, the Alsodes + Eupsophus clade (Anura: Alsodidae), with comments on the supposed paraphyly of Eupsophus

The frog clade composed of the alsodid genera Alsodes + Eupsophus is the most species‐rich of the Patagonian endemic frog clades, including nearly 31 of the slightly more than 50 species of that region. The biology of this group of frogs is poorly known, its taxonomy quite complex (particularly Alsodes), and its diversity in chromosome number striking when compared with other frogs (collectively, there are species having 2n = 22, 2n = 26, 2n = 28, 2n = 30 or 2n = 34). We present a phylogenetic analysis of this Patagonian frog clade based on mitochondrial and nuclear gene sequences. We sequenced five mitochondrial genes (cytochrome b, cytochrome oxidase I, 12S, 16S, NADH dehydrogenase subunit 1) with three intervening tRNAs, and fragments of three nuclear genes (seven in absentia homolog 1, rhodopsin exon 1, RAG‐1), for a maximum of 6510 bp for multiple specimens from 26 of the 31 species. We recovered Eupsophus as polyphyletic, with E. antartandicus, E. sylvaticus, and E. taeniatus in Batrachylidae, in accordance with most previous hypotheses. Based on this result, we transfer E. antartandicus and E. taeniatus back to Batrachyla, and E. sylvaticus to Hylorina (resurrected from the synonymy of Eupsophus), remediating the paraphyly of Eupsophus. Our results strongly corroborate the monophyly of Alsodes + Eupsophus (sensu stricto), the individual monophyly of these genera, and the monophyly of the species groups of Eupsophus. They also show the non‐monophyly of all non‐monotypic species groups of Alsodes proposed in the past. Our results expose several taxonomic problems particularly in Alsodes, and to a lesser extent in Eupsophus. This phylogenetic context suggests a rich evolutionary history of karyotypic diversification in the clade, in part corroborating previous hypotheses. In Alsodes, we predict three independent transformations of chromosome number from the plesiomorphic 2n = 26. All these, strikingly, involve increments or reductions of pairs of haploid chromosomes. Finally, the phylogenetic pattern recovered for Alsodes and Eupsophus suggests a trans‐Andean origin and diversification of the group, with multiple, independent ingressions over cis‐Andean regions.

New parental care behaviours in two telmatobiine genera from temperate Patagonian forests: Batrachyla and Eupsophus (Anura: Leptodactylidae)

Parental care behaviour is described for the first time for two frog species of Batrachyla and three species of Eupsophus , from the Patagonian forests of Chile and Argentina. Males of Batrachyla species attend to eggs which are hidden under shelters on damp soil. Males of Eupsophus species remain by the clutch, and later on also by the tadpoles, in small aquatic microhabitats in the ground. The fact that some male of Eupsophus were found in advanced stages of weight loss confirms the hypothesis that they remain by the clutch until metamorphosis takes place. The parental care behaviour observed in Eupsophus and Batrachyla complements the reproductive strategy of their species, which have few eggs relatively rich in yolk and conceal the clutch in semi-terrestrial environments, and in Eupsophus , have unpigmented endotrophic tadpoles which remain confined to a dark, aquatic microhabitat until their metamorphosis is complete.

Genetic diversity and population structure of two South American marine gastropods, Crepipatella dilatata and C. fecunda (Gastropoda: Calyptraeidae): Distinct patterns based on developmental mode

Abstract We have linked phylogenetics with population genetics in order to investigate the effects of development mode on the genetic diversity and population structure of two southern South American marine gastropod species, Crepipatella dilatata and C. fecunda. DNA sequences were obtained for the mitochondrial gene cytochrome oxidase (COI). We have found that the mean level of genetic diversity was significantly greater in C. dilatata than in C. fecunda, the latter having the highest number of shared haplotypes. The AMOVA analysis shows that C. dilatata has a marked population genetic structure, with 56.86% of the observed genetic variation explained by the variation between populations, in contrast to the genetic homogeneity exhibited by C. fecunda in which most of the variance (97.33%) is explained by the intrapopulational variation. For C. dilatata the multimodal curve obtained is consistent with a long-term stationary population model, whereas in the case of C. fecunda, the mismatch distribution shows a significant fit to a population growth model. Both, the level of genetic differentiation among populations as well as the patterns of genetic structure, correlate well with the larval developmental mode. The role of the environment, selection and historical events on the level of genetic differentiation and the population genetic structure are discussed for both species.

Alsodes Australis, A New Species of Leptodactylid Frog from the Temperate Nothofagus Forest of Southern Chile and Argentina

AbstractA new species of frog, Alsodes australis, is described from the temperate Nothofagus forest of southern Chile and Argentina, based on adults and tadpoles. Up to now, no sympatric occurrence of another congeneric species is known from the type locality. The genus comprises three species groups. From a karyological point of view this species is included in the monticola group, whose species have 26 chromosomes.

Evolutionary history of the Chilean frog genus Telmatobufo (Leptodactylidae): an immunological approach

The quantitative micro-complement fixation technique was used to estimate degrees of relatedness and times of divergence between Caudiverbera and Telmatobufo . The data suggest that the lineages leading to Caudiverbera and Telmatobufo diverged about 35 million years ago. The divergence time among the three species of Telmatobufo was estimated to be 20-25 million years. Ouchterlony tests did not show immunological identity among T. australis and T. bullocki with Caudiverbera , while T. venustus showed partial identity. These analyses suggest that two groups can be recognized within the genus Telmatobufo . The molecular weights of albumins were similar among the Telmatobufo species and slightly lower in C. caudiverbera . Se estimo el grado de parentesco y tiempo de divergencia entre Caudiverbera y Telmatobufo a traves del ensayo de microfijacion de complemento. Los resultados sugieren que el linaje que condujo a Caudiverbera y Telmatobufo divergio hace 35 millones de anos. El tiempo de divergencia entre las especies de Telmatobufo se estimo entre 20-25 millones de anos. El ensayo de Ouchterlony no mostro identidad inmunologica entre T. australis y T. bullocki con Caudiverbera , mientras que T. venustus mostro identidad parcial. Mediante estos analisis fue posible reconocer dos grupos dentro de Telmatobufo . Los pesos moleculares de las albuminas fueron similares entre las especies de Telmatobufo y un poco mas bajo en Caudiverbera .

Relaciones filogenéticas de Rhodolirium montanum Phil. y especies afines, basadas en secuencias nucleotídicas de la región ITS y análisis cariotípico

La taxonomia de los generos chilenos de Amaryllidaceae es confusa, persistiendo problemas taxonomicos tanto a nivel de generos como de especies. Actualmente existe controversia acerca de la correcta clasificacion de especies hasta ahora asignadas a Rhodophiala C.Presl. A traves del estudio de aspectos morfologicos, recientemente se ha sugerido la rehabilitacion del genero Rhodolirium Phil., donde Rhodolirium montanum Phil. es considerada como especie tipo. Esta especie ha sido denominada Rhodophiala rhodolirion (Baker) Traub en las clasificaciones tradicionales. Dado que los datos morfologicos han provisto informacion limitada y ambigua en la clasificacion de las Amarilidaceas chilenas, este trabajo evalua la pertinencia de formalizar la rehabilitacion del genero Rhodolirium y en particular de la especie Rhodolirium montanum, a traves de un analisis de morfologia cromosomica y de secuencias nucleotidicas de la region ITS de seis especies nativas de Amaryllidaceae. Los datos moleculares indican que las especies Rhodophiala bagnoldii, R. montana, R. splendens, R. phycelloides y R. ananuca forman un grupo monofiletico mas emparentado con Hippeastrum Herb. que con Rhodolirium, genero que se presenta como grupo hermano de Phycella en el analisis filogenetico. La separacion de Rhodolirium con respecto de Rhodophiala esta respaldada por diferencias en numero y morfologia cromosomica, presentando el primero un numero cromosomico de 2n = 16 y un indice de asimetria intracromosomica (Isc) de 0,46; mientras que el segundo posee 2n = 18 e Isc de 0,59-0,62. De esta manera, los resultados de los estudios cromosomicos y moleculares, junto a diferencias en exomorfologia, tales como forma del estigma, perigonio y paraperigonio, apoyan la validez del genero Rhodolirium Phil. y por ende de la especie Rhodolirium montanum Phil.

Testing species boundaries between Atlantic and Pacific lineages of the Patagonian rockfish Sebastes oculatus (Teleostei: Scorpaenidae) through mitochondrial DNA sequences

Se investigo la diferenciacion genetica y limites de especies entre los linajes Atlantico y Pacifico de Sebastes oculatus a traves de secuencias de la region D-loop del ADN mitocondrial (541 bp). Las secuencias se obtuvieron de 47 individuos de siete localidades de las costas del Pacifico y Atlantico de Sudamerica (S. oculatus) y una localidad de Sudafrica (S. capensis). Adicionalmente, se obtuvieron dos secuencias de Helicolenus lengerichi que fueron utilizadas como grupo externo. Estos datos fueron combinados con secuencias disponibles en GenBank correspondientes a 21 especies de Sebastes. Las aproximaciones de maxima verosimilitud y analisis Bayesiano mostraron distintividad topologica entre las poblaciones sudamericanas y africanas de Sebastes, apoyando la existencia de dos especies filogeneticas: S. oculatus y S. capensis. Sin embargo, las poblaciones del Pacifico y del Atlantico de S. oculatus no formaron grupos monofileticos reciprocos. La aplicacion del protocolo de Wiens & Penkrot para evaluar limites entre especies no apoyo la existencia de dos especies en las costas de Sudamerica. El flujo genico entre las poblaciones de S. oculatus podria ser explicado por una extensa dispersion larval favorecida tanto por la corriente de Humboldt y la Corriente de la Deriva del Oeste a lo largo de las costas de Sudamerica.

Catastro de vertebrados terrestres y análisis en seis habitats presentes en la Reserva Nacional Futaleufu, provincia de Palena, X Región, Chile

Se realizo un catastro de los mamiferos, aves, reptiles y anfibios presentes en seis habitats de la Reserva Nacional Futaleufu, provincia de Palena, Chile. Estos habitats se encuentran desde la cota 700 hasta los 2000 msnm, ellos son: cipresal, matorral mixto, bosque de lenga y coigue, renoval de Nothofagus, mallin y bosque achaparrado de nirres en sectores altos de roquerios con nieves estacionales. Se registraron un total de 60 especies de vertebrados en la Reserva: 17 especies de mamiferos, 38 especies de aves, cuatro especies de anfibios y una especie de reptil. Se destaca la presencia de huemules en casi todos los habitats. El analisis por habitat muestra una mayor riqueza de especies en los bosques de lenga-coigue, los cuales presentan ademas una mayor heterogeneidad estructural. El matorral mixto, que limita con praderas y campos agricolas, y los renovales de Nothofagus representan el segundo y tercer habitat con mayor riqueza. Se advierte la fragilidad del sistema para mantener poblaciones de vertebrados, pues hacia el borde de la Reserva se produce una interfase entre el uso por animales domesticos (perros, ganado y caballos) y la fauna silvestre endemica o con problemas de conservacion.

Osteología, taxonomía y relaciones filogenéticas de las ranas del género Telmatobufo (Leptodactylidae)

Se hizo un analisis osteologico de las especies del genero Telmatobufo (T. australis, T. bullocki, T. venustus), el que completa la definicion generica fundamentada exclusivamente en la osteologia de T. bullocki y caracteres larvarios. Para cada especie se entrega nueva informacion distribucional, ecologica, reproductiva, morfometrica, cromosomica, alozimica y molecular. Se presenta una clave para identificar individuos adultos de las tres especies de Telmatobufo. Se publica por primera vez una lamina de Philippi perdida por casi un siglo, lo que permite aclarar de manera definitiva la identidad taxonomica de T. venustus. Con las distancias de Mahalanobis y las distancias geneticas de Rogers se determinan las similitudes morfologicas y geneticas entre las especies de Telmatobufo. Usando las secuencias para ARN 12S mitocondrial, frecuencias alelicas polarizadas de genes estructurales y caracteres morfologicos (osteologicos y morfologicos externos) se propone una hipotesis de relaciones filogeneticas basada en los principios de maxima parsimonia y maxima probabilidad. Finalmente se entrega una proposicion acerca de la especiacion e historia evolutiva de este grupo de ranas endemicas de los bosques templados del centro-sur de Chile