Tangled banks: A landscape genomic evaluation of Wallace's Riverine barrier hypothesis for three Amazon plant species

Wallace's Riverine Barrier hypothesis is one of the earliest biogeographic explanations for Amazon speciation, but it has rarely been tested in plants. In this study, we used three woody Amazonian plant species to evaluate Wallace's Hypothesis using tools of landscape genomics. We generated unlinked single‐nucleotide polymorphism (SNP) data from the nuclear genomes of 234 individuals (78 for each plant species) across 13 sampling sites along the Rio Branco, Brazil, for Amphirrhox longifolia (8,075 SNPs), Psychotria lupulina (9,501 SNPs) and Passiflora spinosa (14,536 SNPs). Although significantly different migration rates were estimated between species, the population structure data do not support the hypothesis that the Rio Branco—an allopatric barrier for primates and birds—is a significant genetic barrier for Amphirrhox longifolia, Passiflora spinosa or Psychotria lupulina. Overall, we demonstrated that medium‐sized rivers in the Amazon Basin, such as the Rio Branco, are permeable barriers to gene flow for animal‐dispersed and animal‐pollinated plant species.     

Contrasting patterns of genetic structure in Caryocar (Caryocaraceae) congeners from flooded and upland Amazonian forests

In the present study, we compare the genetic structure of a flooded forest tree Caryocar microcarpum and a terra firme forest tree Caryocar villosum in the lower Rio Negro region and test the hypothesis that the Rio Negro, the largest tributary on the left bank of the Amazon River, has been acting as a geographical barrier to gene flow between populations from the left and right banks. Seventeen adult individuals on the left bank and 27 on the right bank of Rio Negro were sampled for C. microcarpum, whereas 27 on the left and 20 on the right bank were sampled for C. villosum. Two chloroplast DNA regions were sequenced: the intron of trnL gene and the intergenic region between psbA and trnH genes; and all individuals were genotyped using ten microsatellite loci. The trnL intron and psbA‐trnH intergenic spacer generated fragments of 459 bp and 424 bp, respectively. For C. microcarpum, six haplotypes were identified for trnL and seven for psbA‐trnH. By contrast, only one haplotype was found for C. villosum for both sequences. The results obtained showed that the Rio Negro has not been a barrier to gene flow by pollen and seeds for either species. No genetic differentiation and a high migration rate between populations from the left and right banks of the Rio Negro were detected for the chloroplast sequences and nuclear microsatellites, for both C. villosum and C. microcarpum. Although the two analysed sequences showed a sharp topology difference, both indicated that multiple lineages may have contributed to the origin of C. microcarpum populations in the Rio Negro basin. Nevertheless, for C. villosum, from terra firme, the results obtained may provide evidence of a recent expansion of one maternal lineage from an ancient relic population surviving in one of the few moist forest refuges of the Guiana Shield during extended droughts of the glacial periods. We hypothesize that the contrasting environments colonized by this congener pair may have played an important role in shaping the genetic structure of both species. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 98 , 278–290.     

Phylogeography and diversification of an Amazonian understorey hummingbird: paraphyly and evidence for widespread cryptic speciation in the Plio‐Pleistocene

Straight‐billed Hermit Phaethornis bourcieri inhabits the understorey of upland terra firme forest throughout most of the Amazon basin. Currently, two allopatric taxa regarded as subspecies are recognized: P. b. bourcieri and P. b. major. However, the validity, interspecific limits and evolutionary history of these taxa are not yet fully elucidated. We use molecular characters to propose a phylogenetic hypothesis for populations and taxa grouped under Phaethornis bourcieri. Our results showed that P. bourcieri is part of the ‘Ametrornis’ clade, along with Phaethornis philippii and Phaethornis koepckeae, and that the subspecies major is more closely related to the latter two species than to populations grouped under nominate bourcieri. Our phylogenetic hypotheses recovered three main reciprocally monophyletic clades under nominate bourcieri separated by the lower Negro River and the Branco River or the Branco–Negro interfluve (clades B and C) and the upper Amazon (Solimões) or lower Marañon/Ucayali Rivers (clades C and D). Based on multi‐locus phylogeographic and population genetics approaches, we show that P. b. major is best treated as a separate species, and that P. b. bourcieri probably includes more than one evolutionary species, whose limits remain uncertain. The diversification of the ‘Ametrornis’ clade (P. bourcieri, P. philippii and P. koepckeae) is centred in the Amazon and appears to be closely linked to the formation of the modern Amazon drainage during the Plio‐Pleistocene.     

Cryptic diversity in the mtDNA of the ornamental fish Carnegiella strigata

Mitochondrial DNA ( mtDNA) sequences of the marbled hatchetfish Carnegiella strigata, an ornamental fish exported from the Negro River, was examined to determine its genetic diversity and population structure in blackwater rivers (Negro and Uatumã Rivers) in the central Amazon. Analyses of a 646 bp fragment of the ATPase 6/8 mtDNA gene revealed two monophyletic lineages of C. strigata with considerable genetic distance between them (10–12%), suggesting that these lineages should not be considered a single stock. Furthermore, there were strong differences in the geographical distribution of the lineages. These results indicate a past association between drainages of the Negro and Uatumã Rivers. They also suggest that, in the Negro River, its main tributary, the Branco River, may act as a geographical barrier and potentially an ecological barrier between populations of the middle and lower portions of the river.     

South American electric knifefishes of the genus Archolaemus (Ostariophysi,Gymnotiformes): undetected diversity in a clade of rheophiles

Neotropical electric knifefishes of the family Sternopygidae previously considered to represent a single relatively geographically widespread and morphologically variable species, Archolaemus blax, were analysed and found to represent a complex of five species, four of which are new to science. A fifth undescribed species from the Rio São Francisco basin outside the previous known range of the genus was identified. Recognized species of Archolaemus are: A. blax, previously thought to occur in the Rio Araguari, Rio Branco, Rio Tapajós, Rio Tocantins, and Rio Xingu, but which instead proved to be endemic to the Rio Tocantins; Archolaemus ferreirai sp. nov. from the Rio Mucajaí and Rio Uraricoera in the north‐eastern portions of the Amazon basin; Archolaemus janeae sp. nov. of the Rio Xingu and the upper Rio Tapajós, both southern tributaries of the mainstream Amazon; Archolaemus luciae sp. nov. of the Rio Trombetas, Rio Jari, and Rio Tapajós basins of the eastern Amazon, and the independent Rio Araguari draining into the Atlantic Ocean north of the mouth of the Amazon; Archolaemus orientalis sp. nov. of the São Francisco basin in eastern Brazil; and Archolaemus santosi sp. nov. of the Rio Jamari in the south‐western portion of the Amazon basin. The phylogenetic placements of Archolaemus and the recently described genus Japigny relative to the other members of the Eigenmanninae are discussed. A series of synapomorphies for Archolaemus are proposed and a hypothesis of the relationships within that genus is advanced. Rheophily of all members of Archolaemus is discussed, with the genus found to be the most specious clade within the Gymnotiformes living primarily in high‐energy settings. The reported anterior projection of the dentary teeth in A. blax was found to be a consequence of postmortem displacement.     

Characterization of microsatellite loci for the Amazonian rummy‐nose tetra,Hemigrammus bleheri (Teleostei,Characidae)

The rummy‐nose tetra, Hemigrammus bleheri, is a small fish found in forest streams of the Rio Negro floodplain, central Amazonia. This species is popular in aquaria and used as an ornamental fishery resource by the riverine people from middle Rio Negro. Here we describe eight microsatellite DNA loci for rummy‐nose tetras. Number of alleles and heterozygosity per locus in a sample of 30 fish ranged from five to 22 and from 0.52 to 0.96, respectively. These highly variable genetic markers provide important tools for investigating population history and identifying conservation units in rummy‐nose tetras.     

Molecular signature of the D-loop in the brown pencilfish Nannostomus eques (Characiformes, Lebiasinidae) reveals at least two evolutionary units in the Rio Negro basin, Brazil

The genetic variability of the brown pencilfish Nannostomus eques was studied, based on an analysis of sequences from the control region (1084 bp) of mitochondrial (mt)DNA in 125 individuals collected from eight tributaries along the upper (Açaituba, Miuá, Jaradi and Arixanã), middle (Demini), and lower (Jacundá, Maguari and Catalão) Rio Negro (Brazil). Phylogenetic inferences using mtDNA data from N. eques revealed two evolutionary units. Genetic distance between them ranged from 5·5 to 8·3% and differed by 8·5–11·8% from the sister species pencilfish Nannostomus unifasciatus. The time of divergence between the two evolutionary units was estimated to be the Middle Pliocene (c. 2·99 million years before present). Population genetic analysis (DNA polymorphism, AMOVA and Mantel test) showed high haplotype diversity (HD, >0·90) in each evolutionary unit, a strong population genetic structure in the Demini River that formed a monophyletic group and a correlation between genetic divergence and geographical distance in only one of these units (evolutionary unit 1). On the basis of molecular data, the rapids and waterfalls near São Gabriel da Cachoeira (Upper Rio Negro) were the main barriers to gene flow within evolutionary unit 1 in some localities. The emergences of the Branco River and the Anavilhanas Archipelago were apparently responsible for the discrepancy in distribution of the two evolutionary units, except at Jacundá, where the evolutionary units were sympatric. In view of the differences between the evolutionary units, N. eques cannot be treated as a single stock in the Rio Negro basin. These results may have important implications for the fishery management of this ornamental fish.     

The potential role of seed banks in the recovery of dune ecosystems after removal of invasive plant species

Question: How resilient is the seed bank of an invaded dune system? Is that resilience dependent on duration of invasion? How does the accumulated litter layer contribute to the soil seed bank? Location: Coastal sand dunes invaded by Acacia longifolia, Portugal. Methods: Seedling emergence was used to quantify and compare soil seed banks in long‐invaded, recently invaded and non‐invaded areas. Changes in seed banks were also compared with areas where A. longifolia and the litter layer were removed. Results: Species richness, seedling density and diversity were higher in non‐invaded and recently‐invaded areas than in long‐invaded areas. Although there was an apparent similarity between non‐invaded and recently‐invaded areas, analyses of species traits revealed differences. Non‐invaded areas had a wider array of traits. Exotic/invasive species dominated invaded seed banks while native species dominated non‐invaded seed banks. Life forms, growth forms, longevity and dispersal mode showed differences between areas, with cleared plots of long‐invaded areas being apparently the most similar to non‐invaded plots. Acacia longifolia seeds were most abundant in long‐invaded areas, particularly where the litter layer remained. Removal of A. longifolia plus the litter had little effect on the seed bank composition of recently‐invaded areas but resulted in noticeable changes in seed banks of long‐invaded areas. Conclusions: Long‐invaded areas are less resilient and show a higher reinvasion potential, despite severe alteration of the seed banks of both areas. Seed bank studies can be a useful tool to guide management, but can give misleading results when invasion periods are protracted.     

Patterns of Genetic Population Differentiation in Four Species of Amazonian Frogs: A Test of the Riverine Barrier Hypothesis1

Patterns and levels of allozyme variation among populations of Amazonian frogs were used to test the riverine barrier hypothesis of species differentiation. Two frog species were sampled from each of the two main forest habitats on both banks of the Juruá River in the southwestern Brazilian Amazon Basin at various points along its course to contrast different barrier strengths. Scarthyla ostinodactyla and Scinax rubra were sampled from flooded forest (varzea), and Physalaemus petersi and Epipedobates femoralis from non-flooded forest (terra firme). All species showed high levels of within-population genetic variation. Average Nei's (1978) and Rogers’ (1972) genetic distances between sampled sites for all species were high indicating substantial among-population differentiation. The observation of low gene flow between sampled sites within species was further substantiated with Slatkin's (1993) M? analyses. Randomization tests suggested that there was some population structure at a few assayed polymorphic loci that was consistent with the riverine barrier hypothesis. However, it was apparent from the raw allozyme frequency data that these results were largely driven by substantial differentiation at one or a few collecting localities rather than by basin-wide patterns of riverine differentiation. Phenograms using genetic distance matrices supported this interpretation. Patterns of geographic variation are probably more consistent with the idea of this region being a zone of secondary contact.     

An alternative hypothesis for the origin of Amazonian bird diversity

Aim To determine the origin of the high diversity of birds and other animals and plants in Amazonia. Previous hypotheses are: palaeogeography hypothesis, river hypothesis, river-refuge hypothesis, refuge hypothesis, disturbance-vicariance hypothesis, gradient hypothesis, pest-pressure hypothesis, intermediate disturbance hypothesis, riverine disturbance hypothesis, models of fine-scale habitat heterogeneity, lake hypothesis, and museum hypothesis. Methods At present there is agreement between areas of high species diversity and sites located over 100 m. As these sites would have been islands during a sea-level rise of this value, it was important to determine the probable distribution of these islands in Amazonia during a marine transgression. For that purpose, I traced the 100 m contour line from topographic maps. Results Two broad marine transgressions would have been produced from the Atlantic Ocean via the Amazon and Orinoco rivers. Two very large islands would have been formed to the north of the Amazon River, and other islands and archipelagos would have been formed along the coastal lowlands of Guiana, and at the periphery of the Amazon basin. The area located between the Solimoýes and Negro rivers and in the lower Branco would have been completely covered by the sea. Main conclusions A substantial part of the high diversity of forest and nonforest birds in lowland Amazonia can be hypothetysed to have originated during sea-level rises of about 100 m in the Quaternary and late Tertiary. These transgressions would have fragmented the Amazonian lowland into a large number of true islands and archipelagos, thus favouring active allopatric speciation. Values appreciably higher than 100 m above the present sea-level during previous periods of the Tertiary would have produced segregation of the biota earlier than the Tertiary–Quaternary boundary. Sea-level rises and vegetational changes (by aridity or cooling) would thus have driven the speciation pump, and local disturbances and other processes, would maintain the diversity.     

Phylogeography and intraspecific genetic variation of prochilodontid fishes endemic to rivers of northern South America

Phylogeography and intraspecific genetic variation were studied in prochilodontids endemic to the Orinoco, Essequibo and Amazon River basins of northern South America. Portions of two protein‐encoding mitochondrial (mt) DNA genes, ND4 and COI, were examined using single‐strand conformational polymorphisms (SSCPs) and nucleotide sequencing. Phylogeographic analysis indicated that the geographically widespread Prochilodus rubrotaeniatus is paraphyletic, with individuals from the Orinoco sharing most recent common ancestry with co‐occurring Prochilodus mariae . A second Prochilodus rubrotaeniatus clade was composed of haplotypes found in the Rio Cuyuní(Essequibo Basin) and tributaries of the Rio Negro (Amazon Basin). Intraspecific genetic analysis suggested that a complex set of processes have influenced patterns of genetic variation in prochilodontid lineages. Prochilodus rubrotaeniatus is monomorphic at both loci in the Rio Negro and probably recently colonized this basin from the Rio Essequibo. Only two of 55 P. mariae exhibited variant haplotypes, and both had resulted from non‐synonymous changes in the ND4 region. These observations were counter to neutral expectation and consistent with the action of natural selection on the mitochondrion. Overall, these analyses implicate vicariance, demography and selection for driving diversification of prochilodontids in northern South America.     

Ridges and rivers: a test of competing hypotheses of Amazonian diversification using a dart-poison frog (Epipedobates femoralis).

Mitochondrial DNA cytochrome b sequence data from a dart-poison frog, Epipedobates femoralis, were used to test two hypotheses of Amazonian diversification: the riverine barrier and the ridge hypotheses. Samples were derived from sites located on both banks of the Rio Juruá and on both sides of the Iquitos Arch in western Amazonia. The phylogeographic structure was inconsistent with predictions of the riverine barrier hypothesis. Haplotypes from opposite river banks did not form monophyletic clades in any of our phylogenetic analyses, nor was the topology within major clades consistent with the riverine hypothesis. Further, the greatest differentiation between paired sites on opposite banks was not at the river mouth where the strongest barrier to gene flow was predicted to occur. The results instead were consistent with the hypothesis that ancient ridges (arches), no longer evident on the landscape, have shaped the phylogeographic relationships of Amazonian taxa. Two robustly supported clades map onto opposite sides of the Iquitos Arch. The mean haplotypic divergence between the two clades, in excess of 12%, suggests that this cladogenic event dates to between five and 15 million years ago. These estimates span a period of major orogenesis in western South America and presumably the formation of these ancient ridges.     

Modeling the Geographical Distribution and Fundamental Niches of <Emphasis Type="Italic">Cacajao</Emphasis> spp. and <Emphasis Type="Italic">Chiropotes israelita</Emphasis> in Northwestern Amazonia via a Maximum Entropy Algorithm

We modeled the geographical distribution of 4 pithecine primate species: brown-backed bearded sakis (Chiropotes israelita) and 3 black uakaris (Cacajao melanocephalus, C. hosomi, and C. ayresi) that inhabit remote regions of western Amazonas, Brazil. We applied a maximum entropy algorithm modeling program (MAXENT) to field data Boubli collected from 1991 to 2007. We used 23 environmental coverage variables to model the distribution of the primates. The layers were related to precipitation, temperature, topography, and ecological bioregions or Ecoregions. The predicted distribution for Cacajao hosomi was strongly associated with the Negro-Branco Moist Forest and Guianan Highlands Moist Forests Ecoregions, and the Worldclim variables Bio3 (isothermality), Bio4 (temperature seasonality) and Bio17 (precipitation of the driest quarter). Cacajao melanocephalus was strongly associated with Japurá/ Solimões-Negro Moist Forests, Caquetá Moist Forests, Purús Várzea Flooded Forests, Rio Negro Campinaranas, and Cordillera Oriental Montane Forests, Ecoregions. Cacajao ayresi was strongly associated with Negro-Branco Moist Forest and Rio Negro Campinarana Ecoregions as well as Worldclim Bio3 (isothermality). Chiropotes israelita was also strongly associated with Worldclim Bio3 (isothermality) followed by the Negro Branco Moist Forests and Guianan Piedmont and Lowland Moist Forests Ecoregion, and to the Guianan Highland moist forests. These results show a great overlap between the bearded saki and 2 black uakaris, Cacajao hosomi and C. ayresi. Given that one cannot attribute the separation between the species in the Rio Negro-Rio Branco interfluvium to the existence of geographical barriers such as rivers, we suggest that the present geographical boundaries and thus coexistence of the 3 pithecines north of the Rio Negro is maintained by competitive exclusion or stochastic events. Until more surveys are conducted, the present geographical distributions of the pithecines and the mechanism maintaining their boundaries in the Rio Negro-Rio Branco interfluvium will remain uncertain. One important contribution of our model is to identify areas of higher probability of occurrence that might be helpful in guiding future survey expeditions and choices of areas for future conservation of pithecines.     

Phylogeography of a flooded forest specialist fish from central Amazonia based on intron DNA: the cardinal tetra Paracheirodon axelrodi

1. Historic and extant landscape structures and environmental conditions are known to influence phylogeographic patterns and population histories in organisms from Amazonia. Recent work suggests that events of the Tertiary Period have played a significant role in generating biodiversity in this exceptionally rich but understudied region. However, species distributions and population structures are also affected by recent environmental and physical changes, especially within highly dynamic ecosystems. 2. The cardinal tetra is a small luminous fish native to the dynamic flooded forest ecosystems of the Rio Negro and Orinoco basins of South America. This very popular aquarium fish represents the single most important species in the ornamental fishery of the Rio Negro, an activity of high socio‐economic value for local riverine communities. Here we use a fine‐scale sampling regime and sequence data from the second intron of the Ribosomal protein S7 (S7 2) to investigate population structure, colonisation history and genealogical relationships in cardinal tetras of the Rio Negro. 3. High levels of S7 2 polymorphisms revealed phylogeographic patterns across several temporal settings that appear associated with the complex dynamics of the region. Our results suggest a long history of isolation and persistence of cardinal tetra populations in the headwaters and upper regions of the Rio Negro and recent events of colonisation within the incipient Rio Negro floodplain. These colonisation events were followed by recent population expansions likely facilitated by the establishment of the extensive Rio Negro floodplain, an anabranching ecosystem of Holocene age. 4. Our reconstruction of the phylogeographic history of cardinal tetras based on S7 2 intron sequence data reflects the complex influence of both geomorphological and climatic events through time in the Rio Negro basin. Our findings also indicate that in order to maintain the cardinal tetra fishery as a sustainable activity for rural Amazonians, the fishery should be restricted to the middle Rio Negro, the region of the basin where population replenishment is more likely to occur.     

Evidence of two lineages of the dengue vector Aedes aegypti in the Brazilian Amazon, based on mitochondrial DNA ND4 gene sequences

Genetic variation was estimated in ten samples populations of Aedes aegypti from the Brazilian Amazon, by using a 380 bp fragment of the mitochocondrial NADH dehydrogenase subunit 4 (ND4) gene. A total of 123 individuals were analyzed, whereby 13 haplotypes were found. Mean genetic diversity was slightly high (h = 0.666 ± 0.029; π = 0.0115 ± 0.0010). Two AMOVA analyses indicated that most of the variation (~70%-72%) occurred within populations. The variation found among and between populations within the groups disclosed lower, but even so, highly significant values. F(ST) values were not significant in most of the comparisons, except for the samples from Pacaraima and Rio Branco. The isolation by distance (IBD) model was not significant (r = 0.2880; p = 0.097) when the samples from Pacaraima and Rio Branco were excluded from the analyses, this indicating that genetic distance is not related to geographic distance. This result may be explained either by passive dispersal patterns (via human migrations and commercial exchange) or be due to the recent expansion of this mosquito in the Brazilian Amazon. Phylogenetic relationship analysis showed two genetically distinct groups (lineages) within the Brazilian Amazon, each sharing haplotypes with populations from West Africa and Asia.     

Avian distribution patterns in the Guiana Shield: implications for the delimitation of Amazonian areas of endemism

Aim To develop a methodology for defining the boundaries of the Guianan area of endemism using complementary approaches that include GIS tools, multivariate statistics and analyses of physical barriers in the distribution patterns of an entire endemic avifauna. As a case study, I used the distribution patterns of lowland terra firme forest birds. Location Guiana Shield, northern South America. Methods I identified Guianan endemics using the ornithological literature, subsequently gathering distributional data for these taxa using mainly museum collections and my own fieldwork in the region. I used these distributional data to map the spatial patterns of endemicity in the region and to compare distributions across taxa. I employed community composition data from 34 localities from throughout the Guiana Shield to identify spatial patterns of clustering using an ordination analysis (non‐metric multidimensional scaling), and to recognize the region’s main biogeographical barriers for birds using Monmonier’s algorithm. Results At least 88 avian taxa are restricted to the terra firme forests of the Guianan area of endemism, which is roughly delimited by the Amazon, Negro and Branco rivers. These large rivers, however, are not the only boundaries. I identified seven additional barriers, including medium‐sized rivers, non‐forested areas and mountains, which also contribute to delimiting the area of endemism. Within the endemic avifauna, I identified three distinct distribution patterns. The ordination analysis shows the presence of two distinct avifaunas within the Guiana Shield. Main conclusions Although the proposed boundaries of the Guianan area of endemism are consistent with previously postulated configurations, this study reveals a more complex delimitation than formerly recognized, highlighting the importance of several landscape features besides large rivers, and the existence of three distinct distributional patterns within one endemic avifauna. The Branco/Negro interfluvium, often included within this area of endemism, actually represents a transition zone between two distinct avifaunas. The longstanding view of the Amazon Basin as a mosaic of parapatric areas of endemism delimited by major rivers appears to be an oversimplification, at least for the Guiana Shield. This finding suggests the need for more rigorous approaches to re‐evaluate the traditional boundaries of such areas.     

Minimum sample sizes for population genomics: an empirical study from an Amazonian plant species

High‐throughput DNA sequencing facilitates the analysis of large portions of the genome in nonmodel organisms, ensuring high accuracy of population genetic parameters. However, empirical studies evaluating the appropriate sample size for these kinds of studies are still scarce. In this study, we use double‐digest restriction‐associated DNA sequencing (ddRADseq) to recover thousands of single nucleotide polymorphisms (SNPs) for two physically isolated populations of Amphirrhox longifolia (Violaceae), a nonmodel plant species for which no reference genome is available. We used resampling techniques to construct simulated populations with a random subset of individuals and SNPs to determine how many individuals and biallelic markers should be sampled for accurate estimates of intra‐ and interpopulation genetic diversity. We identified 3646 and 4900 polymorphic SNPs for the two populations of A. longifolia, respectively. Our simulations show that, overall, a sample size greater than eight individuals has little impact on estimates of genetic diversity within A. longifolia populations, when 1000 SNPs or higher are used. Our results also show that even at a very small sample size (i.e. two individuals), accurate estimates of F_ST can be obtained with a large number of SNPs (≥1500). These results highlight the potential of high‐throughput genomic sequencing approaches to address questions related to evolutionary biology in nonmodel organisms. Furthermore, our findings also provide insights into the optimization of sampling strategies in the era of population genomics.     

Testing Wallace's intuition: water type,reproductive isolation and divergence in an Amazonian fish

Alfred Russel Wallace proposed classifying Amazon rivers based on their colour and clarity: white, black and clear water. Wallace also proposed that black waters could mediate diversification and yield distinct fish species. Here, we bring evidence of speciation mediated by water type in the sailfin tetra (Crenuchus spilurus), a fish whose range encompasses rivers of very distinct hydrochemical conditions. Distribution of the two main lineages concords with Wallace's water types: one restricted to the acidic and nutrient‐poor waters of the Negro River (herein Rio Negro lineage) and a second widespread throughout the remaining of the species’ distribution (herein Amazonas lineage). These lineages occur over a very broad geographical range, suggesting that despite occurring in regions separated by thousands of kilometres, individuals of the distinct lineages fail to occupy each other's habitats, hundreds of metres apart and not separated by physical barrier. Reproductive isolation was assessed in isolated pairs exposed to black‐water conditions. All pairs with at least one individual of the lineage not native to black waters showed significantly lower spawning success, suggesting that the water type affected the fitness and contributed to reproductive isolation. Our results endorse Wallace's intuition and highlight the importance of ecological factors in shaping diversity of the Amazon fish fauna.     

Three new species of Curimatopsis (Characiformes: Curimatidae) from the Amazon basin

Three new species of Curimatopsis are described from major tributaries of the Amazon basin. Curimatopsis guaporensis n. sp., from the Rio Madeira, belongs to the Curimatopsis evelynae clade and can be distinguished by the distinctive shape of the dark blotch on the caudal peduncle and by the position and shape of the nostrils. Curimatopsis pallida n. sp., from the Rio Negro, also related to C. evelynae, is distinguished from all congeners by the complete absence of pigmentation on the lateral surface of the caudal peduncle. Curimatopsis jaci n. sp., apparently endemic to the upper Rio Tapajós, belongs to the Curimatopsis macrolepis clade and differs from all congeners in details of body pigmentation. Meristic and morphometric features supplement diagnoses for the three new species. These species are hypothesized to belong to the two main clades of Curimatopsis on the basis of previous studies of osteology and external morphology and supplement a recent genetic study that revealed several cryptic and yet undescribed species within the genus. An updated identification key to the species of Curimatopsis is also provided.