Leyla Cárdenas

RECOVERY AND GENETIC DIVERSITY OF THE INTERTIDAL KELP LESSONIA NIGRESCENS (PHAEOPHYCEAE) 20 YEARS AFTER EL NIÑO 1982/831

Massive mortality in kelp beds of the Pacific coasts of North and South America was caused by the rise in surface seawater temperature during the El Niño Southern Oscillation (ENSO) event of 1982/83, the strongest in the four and half previous centuries. In northern Chile a stretch of 600 km of coastline showed massive mortality of the intertidal kelp species Lessonia nigrescens Bory, of which only a few individuals managed to survive. Kelps and their associated biodiversity recovered but kelp beds re‐colonization in general was variable in time and space seemingly very slow along northern Chilean coasts. Here we show, effectively, that northward re‐colonization advanced less than 60 km in 20 years. Conversely, kelp beds of the Northern Hemisphere recovered 300 km in only six months after the same ENSO event. Genetic diversity in the two most affected populations of L. nigrescens shows half of the heterozygosity and polymorphism with respect to that observed in six non affected populations. In addition, geographically separated populations seem highly isolated as evidenced by high and significant fixation indices (all FST values over 0.4).

REDUCED GENETIC DIVERSITY AND INCREASED POPULATION DIFFERENTIATION IN PERIPHERAL AND OVERHARVESTED POPULATIONS OF GIGARTINA SKOTTSBERGII (RHODOPHYTA, GIGARTINALES) IN SOUTHERN CHILE1

This study assesses two hypotheses on the genetic diversity of populations of Gigartina skottsbergii Setchell et Gardner (Rhodophyta, Gigartinales) at the border of the species distribution: 1) peripheral populations display a reduced genetic diversity compared with central populations, and 2) genetic differentiation is higher among peripheral than among central populations. Two peripheral and four central populations were sampled along the Chilean coast and 113 haploid individuals were analyzed using 17 random amplification of polymorphic DNA loci. The genetic diversity was estimated by allele diversity (He), allele richness (Â), and the mean pair‐wise differences among multilocus genotypes. All three estimates consistently and significantly indicated a lower genetic diversity within the peripheral than within the central populations. Genetic differentiation between the two peripheral populations was stronger (FST=0.35) than between central populations at similar spatial scales (FST ranging from 0 to 0.25). In addition, it appeared from the distribution of pair‐wise differences that peripheral populations are in demographic expansion after a recent bottleneck. The results are discussed in the specific context of potential overharvesting of these wild populations.

Diversity of Aplochiton Fishes (Galaxiidea) and the Taxonomic Resurrection of A. marinus

Aplochiton is a small genus of galaxiid fishes endemic to Patagonia and the Falkland Islands whose taxonomy is insufficiently resolved. Recent genetic analyses confirmed the existence of only two closely related species, Aplochiton taeniatus and Aplochiton zebra, while a third controversial species, Aplochiton marinus, remained lost to synonymy with A. taeniatus. Using an integrative taxonomy framework, we studied original samples and published sequences from a broad range in western Patagonia and the Falkland Islands, and generated robust species hypotheses based on single-locus (Cytochrome Oxidase subunit I; COI) species-delineation methods and known diagnostic morphological characters analyzed in a multivariate context. Results revealed three distinct evolutionary lineages that morphologically resemble, in important respects, existing nominal species descriptions. Interestingly, the lineage associated with A. marinus was unambiguously identifiable (100% accuracy) both from the genetic and morphological viewpoints. In contrast, the morphology of A. taeniatus and A. zebra overlapped substantially, mainly due to the high variability of A. taeniatus. Discriminant function analysis aided the identification of these species with 83.9% accuracy. Hence, for their unambiguous identification, genetic screening is needed. A. marinus has seldom been documented, and when recorded, it has always been found in sites with clear marine influence. It is possible that only A. marinus preserves a life cycle related to the sea akin to the hypothesized ancestral galaxiid. We did not find evidence of claimed diadromy in A. taeniatus or A. zebra, and, therefore, these should be regarded as freshwater species. Finally, a lack of phylogeographic patterns and overrepresentation of uncommon haplotypes suggested demographic expansions in recent evolutionary time, especially of A. zebra, in line with the hypothesis of large-scale range expansion and lineage spread in western Patagonia.

Microsatellites of Laminaria digitata tested in Lessonia nigrescens: Evaluation and improvement of cross amplification between kelps of two different families

The use of primers designed originally to amplify DNA for one species in a different one can save time and resources, particularly for microsatellite loci. Microsatellite amplification improvements across two kelp families are reported, where loci originally described in Laminaria digitata (Laminariaceae) were tested in Lessonia nigrescens) was observed in two localities affected by massive mortality events. Nei’s distances among five populations presented similar patterns to those of 30 multilocus dominant loci (RAPD) evaluated in the same localities. Although some success might be achieved in cross-species microsatellite amplifications, the strong mutations detected between these two Laminarian families suggests that better results of cross-amplifications should be expected at much lower taxonomic levels. Thus, although more expensive, construction of new gene libraries is strongly recommended.

Viability selection on body size in a non-marine ostracod

One of the most important research topics in evolutionary ecology is body size evolution. Actually, several hypotheses have been proposed to explain the many observed patterns—also known as “rules”—of body size variation in across latitude, temperature, and time. The temperature–size rule (TSR), describes an inverse relationship between body size and temperature. We took advantage of the “natural laboratory” that the crustacean populations at the Chilean altiplano offers, to study the TSR in ostracods. We studied three populations of Limnocythere atacamae that are physically separated by several kilometers, and differ mainly by their permanent thermal regime. We found larger individuals in the hotspring compared to the cold ponds. Also, in the hotspring we found a significant quadratic selection coefficient, suggesting stabilizing selection in this population. The fitness profiles showed stabilizing selection in the hotspring, and positive directional selection in the ponds. Our results suggest the existence of an optimal body size above the population means. This optimal size is apparently attained in the hotspring population. Then, natural selection appears to be promoting a shift in the mean phenotype that, for some reason, is not attained in the cold environments. Genetic slippage and population bottleneck would explain this absence of response to selection.

Phylogenetic Approaches to Delimit Genetic Lineages of the Mytilus Complex of South America: How Many Species are there?

ABSTRACT The aim of the present work is to increase the general knowledge about an economically important mussel species in Chile. Species of Mytilus are present in the southern cone of South America; however, there is still some controversy about species identification of samples from this area. The study herein presented attempts to: (1) corroborate the phylogenetic hypothesis defined for the Mytilus edulis species complex including taxa from a worldwide distribution; (2) evaluate the possible presence of the species Mytilus trossulus along the Chilean coast and determine if M. trossulus hybridizes with the local species; and (3) provide detailed data collected along the Chilean coast to help define the taxonomic status of Mytilus in South America. To this end, exhaustive sampling was conducted; Mytilus was collected from the Chilean coast and from the coasts of Argentina and Uruguay. Phylogenetic analysis and genetic divergence estimators were used to compare 426 Cytochrome oxidase I mitochondrial gene sequences and 190 16S RNA sequences of Mytilus species sampled from around the world. Following this, the time of divergence between northern hemisphere and southern hemisphere clades of Mytilus species was estimated. In addition, neither M. trossulus nor any associated hybrids were found along the Chilean coast. Finally, the identification of samples from the southern cone of South America is discussed including whether the samples should be identified as Mytilus planulatus or Mytilus platensis. In accordance with the taxonomic priority rules and the results presented here, the species identification frequently used in the literature for samples from the Chilean coast (Mytilus chilensis) may cease to be used.

Molecular and Morphological Evidence Demonstrating Two Species of Helicometrina Linton 1910 (Digenea: Opecoelidae) in Northern Chile

Abstract:  The opecoelid Helicometrina nimia Linton, 1910 has been reported from numerous marine fishes along the Pacific and Atlantic coasts of the Americas. Along the Chilean coast, H. nimia is found in fishes belonging to at least 9 families. This surprisingly low host specificity of H. nimia raises question about the correct identification of specimens assigned to this species. Here we evaluate whether H. nimia specimens isolated from sympatric fish species in northern Chile but with different diets and found in different habitats (water column and demersal) are the same species. Our results demonstrate that specimens from the shallow benthic fish Labrisomus philippii (Steindachner) do not correspond to H. nimia but instead belong to a new species of Helicometrina. This species is described and distinguished from H. nimia using morphological descriptions and 2 molecular markers (the cytochrome c oxidase subunit 1 gene and the V4 region of the SSU rRNA gene). The new species Helicometrina labrisomi (Digenea: Opecoelidae), is found in the intestine of L. philippii (Steindachner, 1866) (Pisces: Labrisomidae), a shallow benthic fish that inhabits the northern coast of Chile. We also studied the related Helicometrina nimia Linton, 1910 from the benthopelagic fishes Paralabrax humeralis (Valenciennes, 1828) and Acanthistius pictus (Tschudi, 1846) (Serranidae). The new species differs from H. nimia by a combination of characters that include ovary shape, number of uterine loops, and position of the genital pore. Our results indicate that morphological characteristics, such as body size, extent of the vitellarium, shape of the testes, and cirrus sac size and extent, traditionally used in the taxonomy of Helicometrina are highly variable. In contrast, meristic and morphological characteristics, such as a lobed ovary, the number of uterine loops, dimensions of the pharynx, and the opening of the genital pore, are highly constant.

Morphological and molecular analysis of centropagids from the high Andean plateau (Copepoda: Calanoidea)

In small metazoan invertebrates classical taxonomic analyses can be ambiguous due to the limited number of morphological characters available. This difficulty can yield incorrect estimates of species richness or taxa distribution. The Boeckella genus has been described as the dominant taxon of zooplankton assemblages in the Andean biogeographical region. In this genus, taxonomic classification and delimitation of boundaries between species has long been problematic and controversial. Among South American centropagids Boeckella gracilipes has been regarded as one of the most broadly distributed species, its presence having been reported from Ecuador to Tierra del Fuego. However, in the high Andean plateau some centropagid populations identified as Boeckella gracilipes have also been considered as B. gracilipes titicacae or even identified as a different species, namely Boeckella titicacae. In an attempt to resolve the taxonomic status of the Centropagidae family from the high Andean plateau, we combined traditional and multivariate morphological analyses (integral approach) with the molecular phylogenetic approach. The results obtained allow us to conclude that centropagids collected from the high Andean plateau actually represent a different species, B. titicacae, not B. gracilipes. The phylogenetic reconstruction of the South American Centropagidae family indicated that B. gracilipes represents a sister taxon to B. titicacae. The present study stresses the usefulness of integrating alfa-taxonomy with morphometric and molecular approaches in order to resolve species boundaries, to determine geographical distributions and to investigate evolutionary processes.

Functional insights into the testis transcriptome of the edible sea urchin Loxechinus albus.

The edible sea urchin Loxechinus albus (Molina, 1782) is a keystone species in the littoral benthic systems of the Pacific coast of South America. The international demand for high-quality gonads of this echinoderm has led to an extensive exploitation and decline of its natural populations. Consequently, a more thorough understanding of L. albus gonad development and gametogenesis could provide valuable resources for aquaculture applications, management, conservation and studies about the evolution of functional and structural pathways that underlie the reproductive toolkit of marine invertebrates. Using a high-throughput sequencing technology, we explored the male gonad transcriptome of this highly fecund sea urchin. Through a de novo assembly approach we obtained 42,530 transcripts of which 15,544 (36.6%) had significant alignments to known proteins in public databases. From these transcripts, approximately 73% were functionally annotated allowing the identification of several candidate genes that are likely to play a central role in developmental processes, nutrient reservoir activity, sexual reproduction, gamete generation, meiosis, sex differentiation, sperm motility, male courtship behavior and fertilization. Additionally, comparisons with the male gonad transcriptomes of other echinoderms revealed several conserved orthologous genes, suggesting that similar functional and structural pathways underlie the reproductive development in this group and other marine invertebrates.

Next-generation transcriptome characterization in three Nacella species (Patellogastropoda: Nacellidae) from South America and Antarctica

The southern tip of South America and Antarctica are particularly interesting due to many genera and also species currently sharing between both areas. The genus Nacella (Patellogastropoda: Nacellidae) is distributed in different regions of South America and Antarctica living preferentially on rocks and boulders and grazing on algae, diatoms and bacterial films. We described the transcriptomes of three Nacella species, Nacella concinna (Strebel, 1908), inhabiting the Antarctic Peninsula; Nacella magallanica (Gmelin, 1791), from Patagonia and Nacella clypeater (Lesson, 1831), from central Chile. In total, we obtained over 20,000 contigs with an average length of 583bp. Homologous protein coding genes (PCGs) for mitochondrial genome of the three species were characterized and a database of molecular markers was also generated. This study represents the first publicly available report on pyrosequencing data for patellogastropod species, and provides an important comparative resource for studies in ecophysiology and evolutionary adaptation in marine invertebrate species.