Phylogeography of the sub-Antarctic notothenioid fish Eleginops maclovinus: evidence of population expansion Santiago Guillermo Ceballos, Enrique Pablo

Lessa, Mariela Fernanda Victorio & Daniel Alfredo Fernndez

Marine Biology International Journal on Life in Oceans and Coastal Waters ISSN 0025-3162 Volume 159 Number 3 Mar Biol (2012) 159:499-505 DOI 10.1007/s00227-011-1830-4

1 23

Your article is protected by copyright and all rights are held exclusively by SpringerVerlag. This e-offprint is for personal use only and shall not be self-archived in electronic repositories. If you wish to self-archive your work, please use the accepted authors version for posting to your own website or your institutions repository. You may further deposit the accepted authors version on a funders repository at a funders request, provided it is not made publicly available until 12 months after publication.

1 23

Author's personal copy

Mar Biol (2012) 159:499505 DOI 10.1007/s00227-011-1830-4

ORIGINAL PAPER

Phylogeography of the sub-Antarctic notothenioid Wsh Eleginops maclovinus: evidence of population expansion
Santiago Guillermo Ceballos Enrique Pablo Lessa Mariela Fernanda Victorio Daniel Alfredo Fernndez

Received: 8 June 2011 / Accepted: 20 October 2011 / Published online: 16 November 2011 Springer-Verlag 2011

Abstract Phylogeography studies add insights into the geographic and evolutionary processes that underline the genetic divergence of populations. This work examines the geographic genetic structure of the Patagonian blennie, Eleginops maclovinus, a notothenioid (Perciformes) endemic to South American temperate and sub-Antarctic waters, using mitochondrial DNA cytochrome b sequences. We found 58 haplotypes in the analysis of 261 individual sequences of 833 base pairs in length. Among-population variance was very low (1.62%) and many haplotypes were shared between several populations across the species geographic range. Genetic diVerentiation was not consistent with a simple model of isolation by distance, possibly suggesting a lack of equilibrium between gene Xow and local genetic drift. The analysis of mismatch distributions, neutrality tests, and the Bayesian Skyline Plot showed a pattern consistent with a recent population expansion event that may have taken place during the Middle Pleistocene.

Introduction Eleginops maclovinus (Cuvier and Valenciennes 1830), known as rbalo in Argentina and Chile, is a notothenioid Wsh (Perciformes) of the monotypic family Eleginopidae. This marine Wsh is endemic to the coastal temperate and sub-Antarctic waters of South America with a range along the Atlantic and PaciWc Patagonian coasts from Valparaiso, Chile (33S) (Pequeo 1989) to San Matias Gulf, Argentina (40S) (Cousseau and Perrotta 2000). This eurithermic and euryhaline species that has been described as a protandrous hermaphrodite (Calvo et al. 1992; Brickle et al. 2005; Licandeo et al. 2006) inhabits costal waters, river mouths and estuaries (Gosztonyi 1980). On the Patagonian coast, E. maclovinus is a common Wsh species and an important component of many trophic webs, both as prey (Goodall and Galeazzi 1985) and as predator, feeding mainly on benthic invertebrates such as crustaceans and polychaetes, but also on algae and Wshes (Isla and San Romn 1995; Licandeo et al. 2006; Martin and Bastida 2008; Pequeo et al. 2010). Additionally, E. maclovinus is important for the recreational and small food Wsheries throughout much of its distribution. Despite its ecological and socioeconomic importance, there is no information regarding its population dynamics and genetic structure, which must be understood for proper conservation and management. Many ecological and evolutionary features make E. maclovinus an interesting species for a phylogeographic study. First, it is an ideal organism to study population responses to environmental variation associated with a broad latitudinal range. Second, E. maclovinus is considered the sister group of the Antarctic notothenioids Wshes (Near and Cheng 2008) that dominate the cold shelf waters of Antarctica (Eastman 2005) and provide a classic example of radiation in the absence of competition from most

Communicated by M. I. Taylor. Electronic supplementary material The online version of this article (doi:10.1007/s00227-011-1830-4) contains supplementary material, which is available to authorized users. S. G. Ceballos (&) M. F. Victorio D. A. Fernndez Centro Austral de Investigaciones CientWcas (CADIC), Bernardo A. Houssay 200, cp 9410, Ushuaia, Tierra del Fuego, Argentina e-mail: sgceballos@gmail.com E. P. Lessa Seccin Evolucin, Facultad de Ciencias, Universidad de la Repblica, 4225 Igu, Montevideo, Uruguay

123

Author's personal copy

500 Mar Biol (2012) 159:499505

other Wsh groups (Clarke and Johnston 1996; Eastman 2000). Therefore, this species likely presents several ancestral traits of the notothenioid radiation (Eastman and Lannoo 2008). And third, the continental shelf area where E. maclovinus is distributed nowadays was largely aVected by Quaternary glacial cycles. Events such as ice sheet calving into the ocean, retraction of the sea coast line and decrease in marine water temperature during glacial periods (Clapperton 1993; Rabassa 2008) might have aVected the habitat suitable for E. maclovinus and thus the population size and geographical structure. Demographic changes may leave genetics footprints in populations (Avise 2000) that can, therefore, be linked to climatic cycles. There are a growing number of studies that associate historical demography with climate changes in a variety of organisms from Patagonia such as grasses species (Jakob et al. 2009), rodents (Lessa et al. 2010), fresh water Wshes (Ruzzante et al. 2008; Zemlak et al. 2010) and crabs (Xu et al. 2009). A population genetic study of E. maclovinus would be an interesting contribution in order to generate an integrated scenario of how the biota has responded to historical climates changes in the entire Patagonian region. To our knowledge, there are no phylogeographic studies on a single marine Wsh species along both Atlantic and PaciWc Patagonian coasts. In this work, we report the Wrst data on population genetics of E. maclovinus along its distributional range using mitochondrial DNA (mtDNA) sequences.

Materials and methods Sample collection E. maclovinus individuals (N = 261) were captured using trammel nets and gill nets at nine sites along the latitudinal range on the Atlantic and PaciWc Patagonian coasts: San Antonio Oeste (SAO) (4050S, 6504W), Puerto Madryn (4246S, 6501W), Rada Tilly (4556S, 6732W) Puerto San Julin (4919S, 6742W), Punta Mara (5357S, 6726W), Canal Beagle (5449S 6810W), Puerto Aysn (4522S, 7251W) Puerto Montt (4132S, 7254W) and Concepcin (3644S, 7311W) (Fig. 1). Muscle samples were collected from each individual and preserved in 99% ethanol. DNA extraction, PCR ampliWcation, and sequencing Total DNA extractions were performed with sodium dodecyl sulfate (SDS)-proteinase K-NaCl-alcohol precipitation (modiWed from Miller et al. 1988). Approximately 900 base pairs of the 3 end of the mitochondrial cytochrome b (cytb)

Fig. 1 E. maclovinus were collected at nine sites (triangles) along the coast of Patagonia, South America. The approximated range distribution of the species is represented in the map by the shadow coastal area. Abbreviations: San Antonio Oeste (SAO)

gene were ampliWed by polymerase chain reaction (PCR) from 261 individuals (2136 per sampling site; Table 1). The PCRs used the forward primer MVZ 05 5-CGA AGC TTG ATA TGA AAA ACC ATC GTT-3 (Smith and Patton 1991) and the reverse primer R1negra 5-CCA GTA CTC CTC CAA GTT TGT CGG GG-3, designed from the alignments of notothenioid cytb sequences. Thirty L of PCR reaction mixtures contained 15 L of total DNA (4 g/mL), 1 unit of Taq DNA polymerase (Promega), 1 Taq polymerase buVer, dNTPs (0.2 mM of each), forward and reverse primers (0.3 mM of each) and MgCl2 (2.5 mM). PCR used the following cycling conditions: an initial denaturation of 3 min at 94C, 30 cycles of 30 s of denaturation at 94C, 30 s of annealing at 50C, and 1 min of extension at 72C, and a Wnal extension of 5 min at 72C. PCRs were performed in a 2720 Thermal Cycler (Applied Biosystems). PCR products were sequenced at Macrogen Korea with both PCR primers. Chromatograms were scored and analyzed using BioEdit (Hall 1999).

123

Author's personal copy

Mar Biol (2012) 159:499505 Table 1 Genetic variation at each site Population SAO Pto. Madryn Rada Tilly Pto. San Julin Pta. Mara Canal Beagle Pto. Aysn Pto. Montt Concepcin All pop. n 25 27 25 24 38 36 31 28 27 261 S 16 23 15 20 20 23 21 23 15 65 No. sin 3 1 10 15 7 14 10 17 9 28 k 10 16 12 9 11 16 16 14 13 58 H 0.90 0.96 0.89 0.86 0.87 0.88 0.91 0.87 0.90 0.90 0.0047 0.0054 0.0026 0.0035 0.0036 0.0038 0.0042 0.0039 0.0032 0.0039 501

Beast v1.6.0. The run consisted of 100 million generations sampled every 1,000 generations under the HKY + G model. Samples before the convergence zone were discarded. Arlequin 3.11 was used to Wt the spatial expansion model (Schneider and ExcoYer 1999; ExcoYer 2004), to estimate the relative timing of expansion.

Results Variation among cytochrome b haplotypes The Wnal data set consisted of 261 cytb sequences of 833 bp in length. An alignment of these sequences showed 65 variable sites, including 28 singletons and 37 parsimony-informative sites, which deWned 58 haplotypes (GenBank accession numbers JN010371, JN010428; Table 1; Online Resource 1). Eight of the 65 variables sites were non-synonymous changes that occurred in one (6), two (1) or three (1) individuals. Haplotype diversity was 0.90 for all individuals and ranged from 0.82 to 0.96 across sites. Nucleotide diversity was 0.0039 for all individuals and ranged from 0.0026 to 0.0054 across sites. The majority (28) of unique haplotypes (32) were deWned by single nucleotide changes. Geographic genetic variation No Wxed diVerences were observed between populations, and many haplotypes were shared between several populations across the geographic sampled range (Fig. 2; Online Resource 1). Among-population variance was very low but signiWcant (1.62%; p = 0.019, AMOVA). Pairwise FST

n, sample sizes; S, number of polymorphic sites; No. sin number of singletons, k, number of haplotypes, H, haplotype diversity; and , nucleotide diversity

Data analysis The number of haplotypes, the number of polymorphic sites and molecular diversity indices for cytb haplotypes, such as haplotype diversity (H) and nucleotide diversity ( ), analysis of molecular variance (AMOVA), pairwise FST values, Mantel test, and Tajimas D and Fus FS tests of neutrality were calculated in Arlequin 3.11 (ExcoYer et al. 2005). Observed and expected mismatch distributions under an exponential growth population model were obtained using DNAsp V5.1(Librado and Rozas 2009). A minimum spanning tree of the haplotypes was generated using the medianjoining method (Bandelt et al. 1999) in Network 4.6 (http:// www.fluxus-engineering.com). Species-speciWc mutation rates ( ) for Wve notothenioids species were obtained from calibrations of uncorrelated lognormal relaxed clocks using Beast v1.6.0 (Drummond and Rambaut 2007) under the HKY + I + G derived from JModeltest (Posada 2008) as the most appropriated model of sequence evolution with a data set of eight cyt b sequences: four representative E. maclovinus sequences, one each from the antifreeze glycoprotein (AFGP)-bearing Antarctic notothenioids Dissostichus eleginoides, Notothenia coriiceps and Chionodraco rastropinosus, and one from the early diverging notothenioid Cottoperca trigloides as the outgroup. For the calibration, the most recent common ancestor of the AFGP-bearing Antarctic notothenioids was set to 24 0.5 Mya (Janko et al. 2007; Near 2004) with a normal prior distribution. Parameter estimates were based on posterior probability distribution constructed by sampling the stationary distribution for 10 million generations every 1,000 steps. Historical population sizes of E. maclovinus were estimated using the coalescent-based generalized Bayesian skyline plot (Drummond et al. 2005) as implemented in

Fig. 2 Minimum spanning tree connecting 58 mtDNA haplotypes. The area of each circle is proportional to frequency of the haplotype

123

Author's personal copy

502 Mar Biol (2012) 159:499505

values ranged from 0.00 to 0.085, indicating low to moderate genetic diVerentiation between populations (Hartl and Clark 2007). A Mantel test did not show a signiWcant correlation between geographic distances and FST (r = 0.23, p = 0.096), thus failing to reveal a pattern of isolation by distance. Phylogenetic relationships between haplotypes According to the minimum spanning tree (Fig. 2), haplotypes found in relatively high frequencies are present in many populations, reXecting the limited geographic structure. The Wve most frequent haplotypes are centrally located in the minimum spanning tree, and most of the remaining low-frequency haplotypes are separated from the common haplotypes by one or few mutational steps. This pattern, fairly star-like, probably reXects the imprinting of a past population expansion (Slatkin and Hudson 1991). Test of neutrality and mismatch distribution Overall values for both Tajimas D and Fus FS were negative and signiWcant (D = 1.99, p < 0.01; FS = 25.93, p < 0.001). They were also negative in each population. Tajimas D was signiWcant at Rada Tilly, Puerto San Julin, Beagle Channel and Puerto Montt, whereas Fus FS was signiWcant at Puerto Madryn, Rada Tilly, Beagle Channel, Puerto Aysn, Puerto Montt and Concepcin (Table 2). Negative values of neutrality tests indicate an excess of low-frequency variants in relation to the mutation-drift equilibrium expectations and suggest a recent population expansion (Ramos-Onsins and Rozas 2002). The mismatch distribution of all sequences was unimodal, suggesting that demographic events in the past have forced coalescent events into a narrow time window in line with a history of a

Fig. 3 Observed and expected mismatch distributions under a model of exponential growth for E. maclovinus from the Argentine and Chilean Patagonian coasts. Dashed lines correspond to the bound of a 90% conWdence interval (CI) for the expected mismatch distribution

past population expansion (Slatkin and Hudson 1991). The test of goodness of Wt did not Wnd signiWcant deviation from expectations under the spatial expansion model (sum of squared deviation = 0.001 and p = 0.83; Fig. 3). Mutational rate estimation and historical population sizes It is possible to calculate the start of the population expansion if an estimate of the locus mutation rate per year ( ) is available. We estimated = 0.0054 substitutions per site per million years (95% highest probability density (HPD) 00240.0087). Using this as a prior, Bayesian skyline plots (Fig. 4) estimated that population expansion started roughly 0.125 Mya (broadly, the results indicate that demographic expansion might have started from 0.1 to 0.175 Mya as could be inferred from the 95% HPD regions drawn in Fig. 4) and that the populations rapidly increased to the present size. The spatial expansion model suggested that the expansion was older, occurring 0.283 Mya (95%CI: 0.1470.523 Mya).

Table 2 Values of Tajimas D and Fus FS neutrality tests and corresponding p values Population Tajimas D D SAO Pto. Madryn Rada Tilly Pto. San Julin Pta. Mara Canal Beagle Pto. Aysn Pto. Montt Concepcin All populations 0.25 0.87 1.61 1.67 1.24 1.46 1.15 1.61 1.10 1.99 P 0.42 0.19 0.04 0.02 0.11 0.06 0.12 0.04 0.13 0.001 Fus Fs Fs 0.96 5.79 5.94 1.35 1.64 6.46 6.59 5.29 5.56 25.93 P 0.35 0.01 0.00 0.26 0.25 0.00 0.01 0.01 0.01 0.00

Discussion Populations with continuous distributions would show genetic structure if ongoing or historical processes have limited gene Xow or if they are under diversifying local selection (Avise 2009). Many marine species, which are expected to experience modest physical barriers to gene Xow and high dispersal potential, exhibit low levels of global population diVerentiation (Ward et al. 1994; Waples 1998). Although direct estimates are lacking, E. maclovinus likely has high dispersal capacity at adult, larval and egg stages. Adult vagility is expected to be high, since E. maclovinus adults

123

Author's personal copy

Mar Biol (2012) 159:499505 503

Fig. 4 Historical estimates of eVective population size (Ne) constructed using the Bayesian Skyline Plot based on mitochondrial cytochrome b haplotypes. The y-axis is the product of eVective population size (Ne) and generation length ( ) in a log scale. The median estimate (black line) and 95% HPD limits (gray lines) are shown

reach a maximum length of 80/90 cm (Gosztonyi 1974; Licandeo et al. 2006) and are highly mobile, at least in aquaria (personal observations). In addition, the E. maclovinus metabolic rate is comparable with that of other highly mobile sub-Antarctic teleosts (Vanella and Calvo 2005). As pointed out by Brickle et al. (2005), E. maclovinus is characterized by high fecundity and small pelagic eggs. Brickle et al. (2005) also suggested that, while inshore waters and estuaries are used for foraging, spawning may occur in deeper waters (30100 m). In this case, the small pelagic eggs and early larval stages would be subjected to strong oVshore currents, which would enhance dispersal. Given the extended pelagic phases of many notothenioid Wshes, larval dispersal by marine currents has been suggested for a number of species in which populations genetic studies commonly found non-signiWcant diVerentiation across thousands of kilometers (Matschiner et al. 2009 and references therein). E. maclovinus has been described as a protandrous hermaphrodite (Calvo et al. 1992; Brickle et al. 2005; Licandeo et al. 2006). In such sex-changing species, the naturally skewed sex ratio is expected to reduce eVective population size and increase genetic drift. As a result, Chopelet et al. (2009) hypothesized that sex-changing species would have more genetic structure than gonochoristic species. However, no diVerence in genetic structure was found among reproductive modes. Our results are concordant with those of previous studies on other notothenioids or marine species which are expected to have high gene Xow between populations. The AMOVA revealed low global genetic variance among populations (1.62%), and allele sharing was extensive. In spite of the low global genetic structure, moderate but signiWcant genetic heterogeneity was observed in some pairwise FST values (data not shown). Nearly one-dimensional spatial

systems (as those of species linked to coastal waters, such as E. maclovinus) have a reduced variance in geographical genetic attributes relative to 2-dimensional spatial systems, increasing the chance to detect a pattern of isolation by distance (Slatkin and Barton 1989). Nonetheless, we failed to detect a signiWcant correlation between FST and geographic distance. This result might come from a relatively recent event of demographic expansion coupled with fairly unrestricted gene Xow. Although FST is an excellent measure of genetic diVerentiation among populations (but see Jost 2008 for qualiWcations), inferences on the extent of migration based on FST are based on a number of assumptions (Whitlock and McCauley 1999) that are likely violated by the data in the present study. The estimate of gene Xow from FST is based on equilibrium expectations derived from neutrality theory as applied to idealized models of population structure (Slatkin and Barton 1989; Avise 2004). Our results (mainly the negative values of neutrality tests, the unimodal mismatch distribution, the absence of a pattern of isolation by distance and the event of rapid increase in population size indicated by the Bayesian Skyline Plot) are suggesting that E. maclovinus is far from both mutationdrift and migration-drift equilibrium and that may have only recently invaded the area it occupies. Therefore, the observed weak global population structure could reXect not only the actual level of gene Xow but also the historical connectivity of populations. As a result, FST-based estimates of gene Xow would be, almost certainly, higher than the actual gene Xow. As in any single locus analysis, we cannot rule out departures from strict neutrality as an alternative (or additional) cause of the observed patterns of genetic variation. Population size changes at high latitudes are generally interpreted in relation to quaternary climatic cycles. Patagonia and Tierra del Fuego experienced signiWcant climatic Xuctuations during the Cenozoic, particularly since the Miocene. Pliocene and Pleistocene glaciations were frequent in this region. Glacial climatic episodes lasted long enough for the formation of a single, continuous mountain ice sheet that extended almost 2,500 km, at least between 36S and 56S. This ice sheet almost completely covered the Patagonian Andean ranges and extended over the piedmont areas to the east (and to the current submarine platform south of the Ro Gallegos) and to sea level in the PaciWc side. The Great Patagonian Glaciation (GPG) that represents the maximum expansion of the ice in extra-Andean Patagonia developed sometime between 1.1 and 1 Mya (Rabassa 2008). Following the GPG, there is evidence of four major glacial episodes, post-GPG 1, post-GPG 2, post-GPG 3 and the Last Glaciation (LG) (following Coronato et al. 2004). Post-GPG 1 event would have taken place possibly before the EarlyMiddle Pleistocene boundary (>0.710 Mya). Post-GPG 2 attained

123

Author's personal copy

504 Mar Biol (2012) 159:499505 Acknowledgments This work was founded by grants from the Agencia Nacional de Promocin CientWca y Tecnolgica (PICT906) and Proyectos Federales de Innovacin Productiva 2007 (PFIP 1622/ 07). We would like to specially thank Roberto Licandeo for sampling support. We also thank Juan Federico Ponce (CADIC-CONICET), Mara Natalia Paso Viola (MACN-CONICET), Alejandro DAnatro and Carolina Abud from Universidad de la Repblica, Montevideo, Ururguay, for their constant technical support and advice.

a major glacial advance at 0.260 Mya (Hein et al. 2009) and post-GPG 3 at 0.145 Mya (Kaplan et al. 2005) during the Middle Pleistocene. The LG of the late Pleistocene attained a maximum (LGM) at approximately 0.025 Mya and ended around 0.015 Mya (Rabassa 2008). The estimate of the time of population expansion from spatial expansion model was older than that of the Bayesian Skyline Plot. Nonetheless, the conWdence intervals of both estimates overlap and suggest that the expansion may have taken place during the Middle Pleistocene, clearly before the LGM but after the post-GPG 1. During the preparation of the manuscript, we became aware of a recent work by Matschiner et al. (2011), who have used six calibration points to address the study of notothenioid divergences. Our estimate of the divergence time between E. maclovinus and the Antarctic notothenioid clade was 42.3 Mya. This estimate is in close agreement with the estimate of 42.9 Mya from Matschiner et al. (2011). The timing of this important event in the phylogenetic history of notothenioids aligns with the early opening of the Drake Passage approximately 41 Mya that led to the formation of the Antarctic Circumpolar Current and the subsequent thermal isolation of Antarctica (Scher and Martin 2006). However, time estimates based on molecular clocks should serve only as rough approximations. A northward retraction during glacial phases is generally suggested in species that have been aVected by glaciations in Patagonia (Ruzzante et al. 2008; Lessa et al. 2010). Under this scenario, E. maclovinus may have had at least two separate refugia during glacial phases, one in the PaciWc Ocean and one in the Atlantic Ocean. This scenario is not supported by our data, since there is extensive haplotype sharing between Atlantic and PaciWc populations and no evidence of substantial subdivision. Furthermore, the pairwise FST between the most northern PaciWc population (Concepcin) and the most northern Atlantic population (SAO) was zero. By contrast, the two most northern Atlantic populations (SAO and Madryn) had somewhat distinct signatures of variation. These locations exhibited higher nucleotide diversity (Table 1) and less negative values of both neutrality tests (Table 2) (except for Fus FS at Madryn) than other sites. These patterns suggest that these populations have historically been more stable and could be the evidence of a single refugium at relatively low latitudes on the Atlantic. This scenario of relative stability in the northeastern portion of the range is compatible with the PaciWc coast being most aVected by glacial phases due to glaciers calving into the coast (Clapperton 1993). Future studies with species-speciWc microsatellite markers developed for E. maclovinus (Ceballos et al. 2011) are currently underway to reveal further information on its population dynamics, structure and phylogeography.

References
Avise JC (2000) Phylogeography: the history and formation of species. Harvard University Press, Cambridge Avise J (2004) Molecular markers, natural history and evolution, 2nd edn. Sinauer Associates, Massachusetts Avise JC (2009) Phylogeography: retrospect and prospect. J Biogeogr 36:315 Bandelt HJ, Forster P, Rhl A (1999) Median-joining networks for inferring intraspeciWc phylogenies. Mol Biol Evol 16(1):37 Brickle P, Laptikhovsky V, Arkhipkin A (2005) Reproductive strategy of a primitive temperate notothenioid Eleginops maclovinus. J Fish Biol 66(4):10441059 Calvo J, Morriconi E, Rae GA, San Romn NA (1992) Evidence of protandry in a sub-Antarctic nototheniid Eleginops maclovinus (Cuv. & Val. 1830) from the Beagle Channel, Argentine. J Fish Biol 40:157162 Ceballos SG, Victorio MF, Lyra ML, Fernndez DA (2011) Isolation and characterization of ten microsatellite loci from the Patagonian notothenioid Wsh Eleginops maclovinus. Conserv Genet Resour 3:689691 Chopelet J, Waples RS, Mariani S (2009) Sex change and the genetic structure of marine Wsh populations. Fish Fish 10(3):329343 Clapperton CM (1993) Quaternary geology and geomorphology of South America, vol 779. Elsevier, Amsterdam Clarke A, Johnston IA (1996) Evolution and adaptative radiation of Antarctic Wshes. TREE 11(5):212217 Coronato AMJ, Martnez O, Rabassa J (2004) Glaciations in Argentine Patagonia, Southern South America. In: Ehlers J, Gibbard P (eds) Quaternary glaciations: extent and chronology. Part III: South America, Asia, Africa, Australia and Antarctica. Quaternary book series. Elsevier, Amsterdam, pp 4967 Cousseau MB, Perrotta R (2000) Peces Marinos de Argentina. INIDEP, Mar del Plata Cuvier G, Valenciennes A (1830) Histoire naturelle des poissons, vol 1. F.-G. Levrault Drummond AJ, Rambaut A (2007) BEAST: Bayesian evolutionary analysis by sampling trees. BMC Evolut Biol 7(1):214 Drummond AJ, Rambaut A, Shapiro B, Pybus OG (2005) Bayesian coalescent inference of past population dynamics from molecular sequences. Mol Biol Evol 22(5):1185 Eastman JT (2000) Antarctic Notothenioid Wshes as subjects for research in evolutionary biology. Antarct Sci 12(3):276287 Eastman J (2005) The nature of the diversity of Antarctic Wshes. Polar Biol 28:93107 Eastman JT, Lannoo MJ (2008) Brain and sense organ anatomy and histology of the Falkland Islands mullet, Eleginops maclovinus (Eleginopidae), the sister group of the Antarctic notothenioid Wshes (Perciformes: Notothenioidei). J Morphol 269(1):84103 ExcoYer L (2004) Patterns of DNA sequence diversity and genetic structure after a range expansion: lessons from the inWnite island model. Mol Ecol 13(4):853864 ExcoYer L, Laval G, Schneider S (2005) Arlequin ver. 3.0: an integrated software package for population genetics data analysis. Evol Bioinform Online 1:4750

123

Author's personal copy

Mar Biol (2012) 159:499505 Goodall RNP, Galeazzi AR (1985) A review of the food habits of the small cetaceans of the Antarctic and Sub-Antarctic. Antarctic nutrient cycles and food webs. Springer-Verlag, Berlin Gosztonyi AE (1974) Edad y crecimiento del rbalo Eleginops maclovinus (Osteichthyes, Nototheniidae) en aguas de la ria Deseado y sus adyacencias. Physis 33(86):18 Gosztonyi A (1980) Biologa del rbalo (Eleginops maclovinus Cuv. & Val., 1830). Universidad Nacional de La Plata, La Plata Hall TA (1999) BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 41:9598 Hartl DL, Clark AG (2007) Principles of population genetics. 4th edn. Sinauer associates, Sunderland Hein AS, Hulton NRJ, Dunai TJ, Schnabel C, Kaplan MR, Naylor M, Xu S (2009) Middle Pleistocene glaciation in Patagonia dated by cosmogenic-nuclide measurements on outwash gravels. Earth Planet Sci Lett 286(12):184197 Isla MS, San Romn NA (1995) Alimentacin de Eleginops maclovinus (Pisces, Nototheniidae) en el Canal Beagle, Argentina. Naturalia Patagnica 3:107127 Jakob SS, Martinez-Meyer E, Blattner FR (2009) Phylogeographic analyses and paleodistribution modeling indicate Pleistocene in situ survival of Hordeum species (Poaceae) in southern Patagonia without genetic or spatial restriction. Mol Biol Evol 26(4):907 Janko K, Lecointre G, DeVries A, Couloux A, Cruaud C, Marshall C (2007) Did glacial advances during the Pleistocene inXuence diVerently the demographic histories of benthic and pelagic Antarctic shelf Wshes?Inferences from intraspeciWc mitochondrial and nuclear DNA sequence diversity. BMC Evol Biol 7(1):220 Jost L (2008) GST and its relatives do not measure diVerentiation. Mol Ecol 17(18):40154026 Kaplan MR, Douglass DC, Ackert RP, CaVee MW (2005) Cosmogenic nuclide chronology of pre-last glaciation maximum moraines at Lago Buenos Aires, 46 S. Argentina Quat Res 63:301315 Lessa EP, DEla G, Pardias UFJ (2010) Genetic footprints of late Quaternary climate change in the diversity of Patagonian Fueguian rodents. Mol Ecol 19(15):30313037 Librado P, Rozas J (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25(11):1451 Licandeo RR, Barrientos CA, Gonzlez MA (2006) Age, growth rates, sex change and feeding habits of notothenioid Wsh Eleginops maclovinus from the central-southern Chilean coast. Environ Biol Fish 77:5161 Martin JP, Bastida R (2008) Contribution of benthic communities to the diet of the Patagonian blenny (Eleginops maclovinus) at Ra Deseado (Santa Cruz, Argentina). Lat Am J Aquat Res 36(1):113 Matschiner M, Hanel R, Salzburger W (2009) Gene Xow by larval dispersal in the Antarctic notothenioid Wsh Gobionotothen gibberifrons. Mol Ecol 18(12):25742587 Matschiner M, Hanel R, Salzburger W (2011) On the origin and trigger of the notothenioid adaptive radiation. PLoS ONE 6(4):e18911 Miller SA, Dikes DD, Polesky HF (1988) A simple salting procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 16:215 505 Near TJ (2004) Estimating divergence times of notothenioid Wshes using a fossil-calibrated molecular clock. Antarct Sci 16(1):3744 Near TJ, Cheng CHC (2008) Phylogenetics of notothenioid Wshes (Teleostei: Acanthomorpha): inferences from mitochondrial and nuclear gene sequences. Mol Phylogenet Evol 47(2):832840 Pequeo G (1989) The geographical distribution and taxonomic arrangement of South American Nototheniid Wshes (Osteichthyes, Nototheniidae). Bol Soc Biol Concepci!n 60:183200 Pequeo G, Pavs H, Bertrn C, Vargas-ChacoV L (2010) Seasonal limnetic feeding regime of therobalo Eleginops maclovinus (Valenciennes 1830), in the Valdivia river, Chile. Gayana (Concepc) 74(1):4756 Posada D (2008) jModelTest: phylogenetic model averaging. Mol Biol Evol 25(7):1253 Rabassa J (2008) Late cenozoic glaciations in Patagonia and Tierra del Fuego. Dev Quat Sci 11:151204 Ramos-Onsins SE, Rozas J (2002) Statistical properties of new neutrality tests against population growth. Mol Biol Evol 19(12):2092 Ruzzante DE, Walde SJ, Gosse JC, Cussac VE, Habit E, Zemlak TS, Adams EDM (2008) Climate control on ancestral population dynamics: insight from Patagonian Wsh phylogeography. Mol Ecol 17(9):22342244 Scher HD, Martin EE (2006) Timing and climatic consequences of the opening of Drake Passage. Science 312(5772):428 Schneider S, ExcoYer L (1999) Estimation of past demographic parameters from the distribution of pairwise diVerences when the mutation rates vary among sites: application to human mitochondrial DNA. Genetics 152(3):1079 Slatkin M, Barton NH (1989) A comparison of three indirect methods for estimating average levels of gene Xow. Evolution 43(7):1349 1368 Slatkin M, Hudson RR (1991) Pairwise comparisons of mitochondrial DNA sequences in stable and exponentially growing populations. Genetics 129(2):555562 Smith MF, Patton JL (1991) Variation in mitochondrial cytochrome b sequence in natural populations of South American akodontine rodents (Muridae: Sigmodontinae). Mol Biol Evol 8(1):85 Vanella F, Calvo J (2005) InXuence of temperature, habitat and body mass an routine metabolic rates of sub-antarctic teleosts. Scientia Marina 69(2):317323 Waples RS (1998) Separating the wheat from the chaV: patterns of genetic diVerentiation in high gene Xow species. J Hered 89(5):438 Ward RD, Woodwark M, Skibinski DOF (1994) A comparison of genetic diversity levels in marine, freshwater, and anadromous Wshes. J Fish Biol 44(2):213232 Whitlock MC, McCauley DE (1999) Indirect measures of gene Xow and migration: FST 1/(4Nm + 1). Heredity 82(2):117125 Xu J, Prez Losada M, Jara CG, Crandall KA (2009) Pleistocene glaciation leaves deep signature on the freshwater crab Aegla alacaluW in Chilean Patagonia. Mol Ecol 18(5):904918 Zemlak TS, Habit EM, Walde SJ, Carrea C, Ruzzante DE (2010) Surviving historical Patagonian landscapes and climate: molecular insights from Galaxias maculatus. BMC Evolut Biol 10(1):67

123