Species,subspecies, or color morphs? Reconsidering the taxonomy of <Emphasis Type="Italic">Callicebus</Emphasis> Thomas, 1903 in the Purus–Madeira interfluvium

There have been recent disagreements as to how many taxa of titi monkeys, genus Callicebus, occur in the region between the Purus and Madeira rivers in western Brazilian Amazonia. Three parapatric taxa were proposed for the area: Callicebus caligatus, Callicebus stephennashi, and Callicebus dubius, but the latter has recently been considered a synonym of C. caligatus, even though both form monophyletic groups and are morphologically distinct. We analyzed the geographic variation in the pelage of Callicebus occurring between the Madeira and Purus rivers and concluded that the phenotypes attributed to C. caligatus and C. dubius are not individual morphs, but rather well-marked and geographically restricted varieties. For this reason, we classify Callicebus caligatus as a polytypic species with two subspecies: Callicebus caligatus caligatus and Callicebus caligatus dubius. This classification is corroborated by molecular evidence as well. The morphological and distributional data indicate that Callicebus stephennashi is a hybrid form of C. c. caligatus and C. c. dubius, due to the presence of intermediate characters. Therefore, until more precise locality records are provided and further evidence is presented, we consider Callicebus stephennashi to be a homonym of the two parental forms.     

A New Subspecies of <Emphasis Type="Italic">Saguinus fuscicollis</Emphasis> (Primates,Callitrichidae)

Saddleback tamarins (Saguinus fuscicollis, S. melanoleucus, and S. tripartitus) occur in the upper Amazon, west of the rios Madeira and Mamoré–Guaporé to the Andes. They currently comprise 13 species and subspecies in Bolivia, Brazil, Colombia, Ecuador, and Peru. Here we report on a previously undescribed subspecies of Saguinus fuscicollis from the interfluvium of the lower rios Madeira and Purus in the central Amazon of Brazil. It is a distinct form with a very pronounced mottling on the back giving the appearance of a saddle, a characteristic shared by the 3 species, and the reason for their collective common name. The lack of a white superciliary chevron and its highly mottled ochraceous-dark brown to black saddle that extends from the scapular region to the base of the tail distinguish the new taxon from Saguinus fuscicollis weddelli. The latter characteristic also distinguishes it from another saddleback tamarin (Saguinus fuscicollis ssp.) that has been reported immediately to the south of its range in the same interfluvium. We compare pelage coloration, external characters and craniometric measurements with its geographically closest relatives, Saguinus fuscicollis weddelli and S. f. avilapiresi, and indicate its probable geographic distribution in a small area between the rios Madeira and Purus, just south of the Rio Amazonas.     

Rondon’s Marmoset, <Emphasis Type="Italic">Mico rondoni</Emphasis> sp. n., from Southwestern Brazilian Amazonia

We describe Rondon’s marmoset (Mico rondoni sp. n.) from the Rio Jamari in the Brazilian state of Rondônia and differentiate the species from other Amazonian marmosets on the basis of morphological, genetic, and zoogeographic characteristics. Mico rondoni sp. n. is a typical bare-eared marmoset of predominantly grayish coloration and unpigmented face and cheiridia, which contrasts clearly with its more brownish and pigmented parapatric congeners Mico melanurus and M. nigriceps. Genetically, the new species is unquestionably distinct from Mico emiliae, with which it had been synonimized previously. The geographic range of the new species is defined as the area delimited by the Mamoré, Madeira, and Jiparaná rivers to the west, north, and east, respectively, and the Serra dos Pacaás Novos, to the south, where it is substituted by Mico melanurus. Depending on its exact limits, this range may cover an area of little more than 100,000 km². A characteristic of the ecology of Mico rondoni sp. n. is its range-wide sympatry with the similarly sized tamarin Saguinus fuscicollis weddelli, which may be a key factor determining the rarity or absence of the new species from many areas within its geographic range. Its apparent scarcity may make the new species exceptionally vulnerable to the effects of the intense human colonization occurring throughout much of its known range, and provoke cause for concern for its long-term survival.     

Spatial and temporal variability of light attenuation in large rivers of the Amazon

The light field and its relationship with biogeochemical variables were investigated in the Solimões, Negro, Amazon, Madeira, Uatumã, Trombetas, and Tapajós Rivers. In high suspended sediment rivers, total suspended matter is the primary control on light attenuation (r = 0.8), with colored dissolved organic matter (CDOM) being secondary (r = ?0.6) due to scattering and absorption, respectively. Photosynthetically active radiation was the lowest (<100.0 μmol m^?2 s^?1 at the depth of half Z _1%) and was limited to depths of less than 1.0 m and confined to red light. In low suspended sediment rivers, CDOM is the primary control on light attenuation (r = 0.9). The concentrations of chlorophyll a (Chla) and CDOM cause variations among these rivers. High CDOM rivers, Negro and Uatumã, are depleted (<0.5% of incoming irradiance) of blue and green light at the depth of half Z _1%. The light spectra of low CDOM and higher Chla waters, such as the Tapajós, Uatumã, and Trombetas Rivers at rising water stage, are restricted to green and red wavelengths, and marked by high absorption at 620 and 670 nm, due to the presence of Cyanophyceae.     

A checklist of Lecythidaceae in the upper Madeira River,Rondônia,Brazil with comments on diversity and conservation

The Lecythidaceae or Brazil Nut family are ubiquitous in non-flooded lowland forests of the Amazon, where they are indicative of well-preserved or little-disturbed habitats. A recent checklist of the Brazilian flora reported 10 genera and 119 species for that country, of which 104 are found in the Amazon region. However, the botanical knowledge in many regions of the country remains far from complete. This study aimed to analyze the diversity of Lecythidaceae in the upper Madeira River region in Rondônia, an area where botanical data is scarce. Lecythidaceae collections deposited in the CEN, INPA and RON herbaria were examined, and four field expeditions in the study area were carried out. For that region, we recorded 37 species of Lecythidaceae belonging to 7 genera: Eschweilera (21 species), Couratari (6), Cariniana (3), Gustavia (3), Allantoma (2), Bertholletia (1) and Lecythis (1). Among these species, five are listed as threatened in the IUCN Red List. The total number of species found in the upper Madeira River of Rondônia alone is higher than the 28 species reported to date for the entire state of Rondônia in the Brazilian Flora Checklist (Smith et al., 2015), and adds 13 new records of Lecythidaceae for the state. Considering that this study is restricted to the northern part of the state, our results suggest that the number of Lecythidaceae species in Rondônia is severely underestimated. Our findings highlight the need for more intensive floristic studies in the Amazon region, which should target areas that are botanically unexplored but have high biodiversity potential. Such studies will provide valuable information to support both taxonomic studies and species conservation assessments.     

Genetic structure of cultivated <Emphasis Type="Italic">Zanthoxylum</Emphasis> species investigated with SSR markers

Zanthoxylum is an economically and ecologically important genus of the Rutaceae family, of which Z. bungeanum and Z. armatum have a long history of cultivation in China. However, how the natural processes such as selection and drift and agriculture practices have influenced the genetic variation of cultivated Zanthoxylum species during long-term domestication remains elusive. Herein, we determined the population genetic structure of current widely cultivated Zanthoxylum species, Z. bungeanum and Z. armatum. Microsatellite markers revealed a high level of genetic variation and significant genetic differentiation for both species despite Z. bungeanum showed higher genetic diversity than Z. armatum. AMOVA indicated that most of the genetic variation exists within individuals rather than among provenances for both species. Population structure analyses generated three distinct groups within the entire accessions. All Z. bungeanum accessions were distinguished into two major geographic groups, north and south groups, with Qinling Mountains as the main geographic barrier to gene flow while a significant genetic differentiation was observed between cultivated and wild Z. armatum accessions. Mantel test of Z. bungeanum displayed a significant correlation between genetic and geographic distances within each inferred group but no correlation between genetic and geographic distance was observed when comparing genetic and geographic distances focusing only on pairwise of north vs. south provenances, ruling out the hypothesis that gene flow between north and south provenances followed an isolation-by-distance model. Our research provided a fundamental genetic profile that will improve the conservation and responsible exploitation of the extant germplasm of Zanthoxylum.     

The Evolutionary History and Genetic Diversity of Kinkajous, <Emphasis Type="Italic">Potos flavus</Emphasis> (Carnivora,Procyonidae)

The genus Potos (Procyonidae) is currently recognized as a monotypic genus comprising the single species Potos flavus, the kinkajou. Kinkajous are widely distributed throughout forested habitats of tropical Central and South America, extending from eastern Brazil across central Bolivia, eastern Peru, northern Ecuador, Guianas, Suriname, Venezuela, Colombia, and then through Central America and into western Mexico. The taxonomic history of the species is complex, with seven or eight subspecies historically recognized to acknowledge the phenotypic variation among populations. In this study, the systematics and the evolutionary history of Potos flavus were investigated based on the mitochondrial gene cytochrome b, including specimens from a large range of localities, covering most of the distribution of the species, from central Middle America (Costa Rica and Panama) through South America (Ecuador, Peru, Bolivia, Brazil, Guyana, and French Guiana). Analyses of 30 Potos flavus sequences showed 27 haplotypes that were grouped in five main clades in all phylogenetic analyses. These clades suggested a high geographic structure with specimens from (1) Costa Rica, (2) Guianas and North Brazil, (3) North Peru, (4) Ecuador and Panama, (5a) interfluves Branco-Negro rivers in the Brazilian Amazon, (5b) Eastern Atlantic Forest, (5c) Amazonian lowlands east Negro river including Bolivia, Peru, and West Brazil. Each of these clades differs from 4.5 % to 9.3 % in their genetic distance estimates, which suggests that the specific status of some of these lineages should be reconsidered. Divergence dating and biogeographic analysis suggested that crown Potos diversified in the Miocene-Pliocene in South America, and geographic barriers, such as savannas and rivers, may have played a significant role in the kinkajou diversification.     

Geographic patterns of genetic variation in nuclear and chloroplast genomes of two related oaks (<Emphasis Type="Italic">Quercus aliena</Emphasis> and <Emphasis Type="Italic">Q. serrata</Emphasis>) in Japan: implications for seed and seedling transfer

In this study, we assessed geographic patterns of genetic variations in nuclear and chloroplast genomes of two related native oaks in Japan, Quercus aliena and Q. serrata, in order to facilitate development of genetic guidelines for transfer of planting stocks for each species. A total of 12 populations of Q. aliena and 44 populations of Q. serrata were analyzed in this study. Genotyping of nuclear microsatellites in Q. aliena was done with only nine populations (n = 212) due to limited numbers of individuals in two populations, while all 12 populations (n = 89) were used in sequencing chloroplast DNA (cpDNA). In Q. serrata, 43 populations (n = 1032) were genotyped by nuclear microsatellite markers, while cpDNA of 44 populations (n = 350) was sequenced. As anticipated, geographic patterns detected in the variations of Q. aliena’s nuclear genome and its chloroplast haplotype distribution clearly distinguished northern and southern groups of populations. However, those of Q. serrata were inconsistent. The geographic distribution of its chloroplast haplotypes tends to show the predicted differentiation between northern and southern lineages, but geographic signals in the genetic structure of its nuclear microsatellites are weak. Therefore, treating northern and southern regions of Japan as genetically distinct transferrable zones for planting stocks is highly warranted for Q. aliena. For Q. serrata, the strong NE-SW geographic structure of cpDNA should be considered.     

Species complex delimitation and patterns of population structure at different geographic scales in Neotropical silver catfish (<Emphasis Type="Italic">Rhamdia</Emphasis>: Heptapteridae)

The Neotropical catfish genus Rhamdia inhabits rivers and lakes from Mexico to Argentina. Previous studies have found that the taxonomy of this genus, as well as that of R. quelen, remains controversial. The present study aims to contribute to the understanding of Rhamdia systematics by delimiting putative species, and to elucidate the pattern of genetic differentiation of Rhamdia at different geographic levels within the cis-andean region. Species boundaries were defined by Generalized Mixed Yule Coalescent and Automatic Barcode Gap Discovery methods, and by phylogenetic analyses of cytochrome b (cyt b) sequences. Moreover, we performed phylogeographic analyses based on cyt b sequences and microsatellite markers. Patterns of differentiation were analyzed at three nested geographic levels: in the main cis-andean basins (macrogeographic scale); in the second major Neotropical basin system (mesogeographic scale), which encompasses La Plata basin, Patos-Merin basin, and the coastal lagoons draining to SW Atlantic Ocean; and finally, in the three most important coastal lagoons for artisanal fisheries in Uruguay (microgeographic scale). Sixteen species were found within Rhamdia, divided into two clades (cis- and trans-andean clades), each composed of eight putative species. Cis-andean Rhamdia species have probably diverged due to vicariance events occurring between and within basins since late Miocene-Pleistocene. Microgeographic scale analysis based on cyt b and microsatellite data revealed a high genetic structuring among the studied coastal lagoons. Mitochondrial and microsatellite markers enabled to identify three different populations, corresponding to the three coastal lagoons analyzed, which would have diverged recently and could be considered as different Management Units.     

A morphological and phylogenetic analysis of <Emphasis Type="Italic">Ornithodoros marinkellei</Emphasis> (Acari: Argasidae), with additional notes on habitat and host usage

Ornithodoros marinkellei was described from larvae collected on Pteronotus spp. bats in Colombia and Panama. More recently, this tick was reported in the Brazilian Amazon. Because some morphometric differences were observed between O. marinkellei larvae from Colombia and Brazil, it was proposed that further investigations were needed to assess whether the differences could be attributed to intra- or inter-specific polymorphism. Herein, we collected O. marinkellei specimens in the type locality of Colombia, in Brazil, and in a new locality in Nicaragua, expanding the distribution of the species to Nicaragua. Morphometric analysis of larvae and adults, corroborated by a principal component analysis (PCA), indicated that the Brazilian specimens were larger than specimens from Colombia and Nicaragua. Phylogenetic analysis inferred from the mitochondrial 16S rRNA gene showed ticks from Colombia and Nicaragua more genetically related than any of them with ticks from Brazil, although ticks from the three countries grouped in a clade sister to a major clade containing sequences of various Neotropical Ornithodoros species. We concluded that ticks identified as O. marinkellei from Colombia, Nicaragua, and Brazil represent the same taxon, and that the genetic and morphological differences between them are likely to have a geographical bias. We redescribed the nymph of O. marinkellei, which has a vestigial hypostome, probably incompatible with blood feeding. We also report human infestation by O. marinkellei adults. As all reports of O. marinkellei adults have been from hot caves (temperature?>?35 °C), this abiotic condition could be a limiting factor for the occurrence of this tick species.     

Genetic structure analysis of mantis shrimp <Emphasis Type="Italic">Oratosquilla oratoria</Emphasis> based on mitochondrial DNA control region sequence

The genetic relationships of mantis shrimp Oratosquilla oratoria between the coastal waters of China and Japan were not well studied. To reveal the genetic differentiation and genetic structure among populations, we collected populations of mantis shrimp O. oratoria from the coastal waters of China and Japan to analyze the mtDNA control region variation. A total of 309 individuals of O. oratoria were collected from 13 localities (11 from China and 2 from Japan) and a segment of mitochondrial DNA control region was sequenced. Three hundred nine haplotypes were defined, yielding a very high haplotype diversity and nucleotide diversity. Two lineages of O. oratoria were revealed and displayed strong differences in the geographical distribution. In the coastal waters of China, the geographic distribution of the two lineages was completely separated by the Yangtze River estuary; however, the lineages showed geographic sympatry in two populations from Japan. Based on the lineage distribution, three groups were defined. There was no significant genetic differentiation among the populations within the three groups, indicating high gene flow within each group. Significant and negative values for Tajima D and Fu’s Fs tests, and mismatch distributions for two lineages indicated population expansion. The present result confirmed that the freshwater outflow from the Yangtze River formed a physical barrier and affected gene exchange. The different distribution patterns of the two lineages in coastal waters of China and Japan indicated that the larvae of O. oratoria were transferred from China to the coastal waters of Japan with a one-way gene flow.     

Modern State of Populations of Endemic <Emphasis Type="Italic">Oxytropis</Emphasis> Species from Baikal Siberia and Their Phylogenetic Relationships Based on Chloroplast DNA Markers

The genetic diversity and population structure of the endemic species of Baikal Siberia Oxytropis triphylla, O. bargusinensis, and O. interposita were studied for the first time on the basis of the nucleotide polymorphism of intergenic spacers psbA–trnH, trnL–trnF, and trnS–trnG of chloroplast DNA. All populations of these species were characterized by a high haplotype (0.762–0.924) and relatively low nucleotide (0.0011–0.0022) diversity. Analysis of the distribution of variability in O. triphylla and O. bargusinensis showed that there was no significant genetic differentiation between populations of each species; the gene flow was 4.43 and 8.91, respectively. The high level of genetic diversity in the studied populations indicates a relatively stable state of these populations. A study of the phylogenetic relationships of closely related species confirms the concept of the origin of O. bargusinensis and O. tompudae as a result of intersectional hybridization of the species of the sections Orobia and Verticillares.     

Morphogenesis,phylogenetic relationships,and distribution of the bryozoans of the genus <Emphasis Type="Italic">Orbipora</Emphasis> Eichwald, 1856 (order Trepostomida)

Five species of the genus Orbipora, including two new species, O. lynnensis sp. nov. and O. ukhakuensis sp. nov., are described from the Middle Ordovician of the Leningrad Region and Estonia. The colonies of the species O. acanthophora Bassler, 1911 and O. solida Bassler, 1911 are measured and figured for the first time. The stratigraphic range and geographic distribution of O. acanthophora are refined. A scheme of possible phylogenetic relationships and distribution maps of the species of the genus are presented.     

Comparative phylogeography of diadromous and freshwater daces of the genus <Emphasis Type="Italic">Tribolodon</Emphasis> (Cyprinidae)

Far Eastern daces, genus Tribolodon (Cyprinidae), are thought to have diversified and developed unique diadromous life histories under changing conditions in the Sea of Japan and the surrounding environment. To examine the relationships between life history traits, distribution, and genetic population structures, we conducted a comparative phylogeographic analysis using partial mtDNA sequence data from samples collected over almost the full ranges of all four Tribolodon species. Phylogenetic analyses revealed several intraspecific haplotype groups that differentiated in the early Pleistocene to the Pliocene with or without geographic overlaps. A time-calibrated phylogeny suggested that the relatively smaller geographic ranges of the strictly freshwater species, T. sachalinensis and T. nakamurai, were explained not by the recent origins of these species, but by their limited dispersal abilities and smaller historical population sizes. The wider-ranging diadromous species, T. brandtii and T. hakonensis, exhibited similar major phylogeographic structures in their distributions, but the chronological order and timing of formation of this structure largely differed between the two species. In addition to those differences, the overlapping patterns of the differentiated intraspecific lineages in these species suggest dynamic, but somewhat restricted dispersal during the Plio-Pleistocene. Tribolodon hakonensis, one of the most widespread species of East Asian freshwater fishes, included both common and unique phylogeographic patterns compared to other fish species; the unique patterns (i.e., its wide range across freshwater biogeographic boundaries like the sea and mountains) would reflect its ecological features as a remarkable generalist inhabiting lakes, upper and lower reaches of rivers, and even coastal areas.     

Delimitation of evolutionary units in Cuvier’s dwarf caiman, <Emphasis Type="Italic">Paleosuchus palpebrosus</Emphasis> (Cuvier, 1807): insights from conservation of a broadly distributed species

An important goal of evolutionary and conservation biology is the identification of units below the species level, such as Evolutionarily Significant Units (ESUs), providing objectively delimited units for species conservation and management. In this study we tested the hypothesis that Cuvier’s dwarf caiman (Paleosuchus palpebrosus)—a species broadly distributed across several biomes and watersheds of South America—is comprised of different ESUs. We analyzed mitochondrial cytochrome b sequences of 206 individuals and 532 unlinked ddRAD loci of 20 individuals chosen from amongst the mitochondrial haplogroups. Analysis of the cytochrome b sequences revealed four mitochondrial clusters, while STRUCTURE analysis of ddRAD loci detected three genomic clusters with different levels of mixture between them. Using the Adaptive Evolutionary Conservation (AEC) framework we identified three ESUs: “Amazon”, “Madeira-Bolivia” and “Pantanal”; one of them composed of two different Management Units (MUs), “Madeira” and “Bolivia”. In general, based on the comparisons with other crocodilian species, genetic diversity of each lineage was moderate however, the Madeira MU showed fivefold lower genetic diversity than other geographic groups. Considering the particularities of each Paleosuchus palpebrosus conservation unit, we recommend that the conservation status of each is evaluated separately. Tropical biodiversity is largely underestimated and in this context the broadly distributed species are the most likely candidates to harbor distinct evolutionary lineages. Thus, we suggest that conservation research should not neglect species that are generally considered of Least Concern by IUCN due to the taxon’s broad geographic distribution.     

On the taxonomic rank of <Emphasis Type="Italic">ciscaucasicus</Emphasis> and its relationships with the pygmy wood mouse <Emphasis Type="Italic">Sylvaemus uralensis</Emphasis> inferred from the mtDNA cytochrome b gene sequence

To specify the taxonomic rank of form ciscaucasicus (independent species Sylvaemus ciscaucasicus, or intraspecific form of pygmy wood mouse, S. uralensis), a 402-bp the mtDNA cytochrome b gene fragment (402 bp) was examined in ciscaucasicus individuals from six geographic localities of the Caucasus and Ciscaucasus (Krasnodar krai and Adygeya Republic) and 17 S. uralensis individuals from seven localities of the Russian Plain (Saratov oblast, Smolensk oblast, Voronezh oblast, Tula oblast, Moscow oblast, Tver’ oblast, and northern Krasnodar krai). For comparison, the cytochrome b gene was partly sequenced in the samples of yellow necked, S. flavicollis (n = 2, Samara oblast), and Caucasian, S. ponticus (n = 6, Krasnodar krai), wood mice. One Mus musculus specimen from Western Europe, whose nucleotide sequences were deposed in the GenBank, was used as intergeneric outgroup. Phylogenetic trees for the forms examined were constructed based on the mtDNA sequence variation and using the neighbor joining and maximum parsimony methods. The network of the cytochrome b haplotypes was also constructed. The level of genetic divergence was evaluated using Kimura’s two-parameter algorithm. Based on the data on the sequence variation in a 402-bp mtDNA cytochrome b gene fragment, the hypothesis on the species status of the ciscaucasicus form was. The mean intergroup distances (d) between the geographic groups of S. uralensis varied from 0.0036 to 0.0152. At the same time, the distances between the pygmy wood mice and the group of S. flavicollis-S. ponticus varies in the range from 0.0860 to 0.0935, and the level of intergeneric genetic differentiation (Sylvaemus-Mus) is higher than the latter index (d = 0.142). Ciscaucasicus should be considered as geographic substitution form of S. uralensis. Furthermore, the Caucasian populations of S. uralensis (= ciscaucasicus) were characterized by a threefold lower value of intergroup genetic divergence (d = 0.0062) than the East European populations (d = 0.0179). This finding pointed to some isolation of Caucasian populations of pygmy wood mouse and depletion of their gene pool. However other molecular genetic data (similarity of nucleotide composition and consistence of the levels of intra-and intergroup distances) suggest the absence of geographic subdivision between Caucasian and East European populations of S. uralensis relative to the molecular marker examined.     

Conservation genetics of the highly endangered Azorean endemics <Emphasis Type="Italic">Euphrasia azorica</Emphasis> and <Emphasis Type="Italic">Euphrasia grandiflora</Emphasis> using new SSR data

In the Azores Islands, two Euphrasia L. (Orobanchaceae) endemic species are recognized: Euphrasia azorica H.C.Watson, an annual herb, in Flores and Corvo, and Euphrasia grandiflora Hochst. ex Seub., a semi-shrub, in Pico, São Jorge and Terceira. Both species are highly endangered and protected by the Bern Convention and Habitats Directive. A population genetics study was conducted with new microsatellite primer pairs in 159 individuals of E. azorica and E. grandifolia, sampled from populations in Flores, Corvo, Pico and São Jorge. Allele sizing suggested that E. azorica is a diploid while E. grandiflora is a tetraploid. Euphrasia grandiflora revealed higher genetic diversity then E. azorica. The E. grandiflora population of Morro Pelado in São Jorge, displayed higher genetic diversity when compared with all others, while the E. azorica population of Madeira Seca in Corvo, showed the lowest. Private and less common bands were also overall higher in E. grandiflora populations. Population genetic structure analysis confirmed a distinctiveness between the two Azorean endemic Euphrasia, in addition to island-specific genetic patterns in E. azorica. The genetic structure obtained for E. grandiflora was complex with the populations of Cabeço do Mistério in Pico Island and of Pico da Esperança in São Jorge sharing the same genetic group, while a putative spatial barrier to gene flow was still retrieved between both islands. Although some populations of both species might benefit from propagation actions, studies are needed on plant host species and translocations between islands or between some populations of a same island should be avoided, due to the occurrence of putative ESUs. Eradication of invasive species and control of grazing will be fundamental to promote in situ restauration.     

Time-calibrated molecular phylogeny reveals a Miocene–Pliocene diversification in the Amazon miniature killifish genus <Emphasis Type="Italic">Fluviphylax</Emphasis> (Cyprinodontiformes: Cyprinodontoidei)

Species of Fluviphylax are widely distributed over the Amazon and Orinoco river drainages and are among the smallest fish in the neotropics, inhabiting areas near the margin of slow-flowing clear and black water streams and lakes. Here, we present the first multigene molecular phylogeny of Fluviphylax, including all five nominal species of Fluviphylax and three undescribed species. The analysis included fragments of one mitochondrial and five nuclear genes, totaling 5880 bp. The dataset was analyzed using maximum parsimony, maximum likelihood, and Bayesian inference approaches providing high-supported well-solved trees. A time-calibrated analysis was performed providing information on the origin and diversification of the miniature genus in the Amazon. We estimate that Fluviphylax lineage splits from its sister group, the Anablepidae and Poeciliidae (Poeciliinae sensu Parenti, 1981), during the Late Eocene, about 36.6 Mya; but lineage diversification started only in the Middle Miocene, about 16 Mya, during the formation of the Pebas system. Subsequent splits within Fluviphylax occurred in the Late Miocene–Pliocene, between 10 and 6 Mya and during the Pliocene, and were probably influenced by paleogeographical events such as the breaching of the Purus arch, the rise of the Vaupés arch, the uplift of the Fitzcarrald arch, and the capture of the Contigo and Uraricoera river drainages by the Branco River. The present time-calibrated analysis provides the first insight on the evolution of one of the smallest vertebrate taxa in the Amazon and Orinoco river drainages.     

Allozyme variability in populations of trout (<Emphasis Type="Italic">Salmo trutta</Emphasis>) from the rivers of Iran

For the first time, an analysis was carried out of allozyme variability in trout (Salmo trutta) from three rivers of Iran. We studied 23 gene loci coding enzymes: glycerol-3-phosphate dehydrogenase (G3PDH), aspartate aminotransferase (AAT), malate dehydrogenase (MDH), lactate dehydrogenase (LDH), creatine kinase (CK), malic enzyme [NADP-dependent MDH] (MEP), superoxide dismutase (SOD), esterase (EST), and esterase D (EST-D). The obtained data demonstrate the similarity between the trout samples from different rivers of Iran according to genetic characteristics. Taking into account the differences by allozyme markers of allele frequencies and allele composition of some loci, we should expect that Iranian trout diverges significantly in genetics from the other trout populations of the Caspian Sea.     

RAPD-PCR Analysis of Ground Squirrels from the Tobol-Ishim Interfluve: Evidence for Interspecific Hybridization between Ground Squirrel Species <Emphasis Type="Italic">Spermophilus major</Emphasis> and <Emphasis Type="Italic">S. erythrogenys</Emphasis>

Populations of two ground squirrel species, Spermophilus major and S. erythrogenys, from the interfluvial area of the Tobol and Ishim rivers, where their ranges overlap, have been examined using RAPD-PCR. We have identified 253 loci, which included taxon-specific markers for S. major and S. erythrogenys as well as markers for geographic populations. Estimation of genetic diversity and construction of phylogenetic relationships were performed using software programs POPGENE, TEPGA, and TREECON. In all, based on morphological traits, animals from the Tobol-Ishim interfluve were assigned to the two parental morphotypes and showed similar levels of genetic variability (H, n_a, n_e). However, the total polymorphism level proved to be higher in ground squirrels with the major morphotype (P = 40.32%,P₉₅ = 27.27%) than in animals with the erythrogenys morphotype (P = 32%,P₉₅ = 22.13%). Nevertheless, the number of rare alleles was high in both cases, constituting about 70% of the total number. Interpopulation differentiation was considerably higher in S. major δ = 0.50) than in S. erythrogenys δ = 0.41). The genetic differentiation between local samples from the Tobol-Ishim interfluvial area was lower than that between the parental species. A significant part of the genetic diversity of the species examined and animals from the zone of overlapping ranges was accounted for by intrapopulation variability. Animals from the northern and southern parts of the Tobol-Ishim interfluve were charac-terized by the core traits of S. major and S. erythrogenys, respectively, falling into two distinct clusters in the UPGMA and NJ reconstructions. In addition to three hybrid individuals, identified by the bioacoustic method, three hybrid animals were distinguished using RAPD analysis. These animals earlier were thought to be “pure” species and formed their own clusters in phylogenetic reconstructions. Thus, the RAPD-PCR results directly showed the existence of stable hybridization (20% genetic hybrids) between S. major and S. erythrogenys in the Tobol-Ishim interfluvial area, which is more extensive than inferred previously from morphological and bioacoustic data.