Genetic divergence and relationships among smelts of the genus Osmerus from the Russian waters

Smelts of the genus Osmerus, O. eperlanus and O. mordax dentes, inhabiting the basins of the Atlantic, Arctic, and Pacific oceans were investigated using RFLP analysis of the mitochondrial DNA segments A8/A6/COIII/ND3 and ND3/ND4, and sequencing of the cytb and COI genes (mtDNA), and intron 1 of the rpS7 gene (nDNA). A total of 14 samples from the populations from most part of the Russian range were examined. The mean values of haplotype and nucleotide diversity constituted 0.5997 +/- 0.11264 and 0.003201 for O. m. dentex, and 0.3086 +/- 0.09892 and 0.000431 for O. eperlanus, respectively. The high level of interspecific diversity (12.94%) along with the low level of intraspercific diversity (0.049% for O. m. dentex, and 0.001% for O. eperlanus was observed. The dendrograms (UPGMA, NJ, MP, and BA) constructed using the data of RFLP analysis of mtDNA, along with the sequencing data of mitochondrial and nuclear genes were congruent. The representatives of O. eperlanus and O. m. dentex formed steady clusters in accordance with their species affiliation, albeit without subdivision into local populations depending on their geographic locality.     

Comparative Analysis of Mitochondrial DNA Diversity in Smelts

The polymerase chain reaction (PCR) was used to amplify a segment of the mitochondrial DNA coding for NADH-dehydrogenase subunits ND5/ND6 in five smelt species (family Osmeridae). Amplified DNA was screened for restriction fragment length polymorphism (RFLP). Nucleotide sequence divergence of mitochondrial DNA between species ranges from 11.9 (between Hypomesus nipponensis and H. japonicus) to 24.7% (between Osmerus mordax dentex and Mallotus villosus catervarius). The genetic divergence between populations of H. nipponensis, H. japonicus, and Osmerus mordax dentex was 0.32, 0.08 to 0.15, and 0.025%, respectively. The absence of common haplotypes enables differentiation of closely related smelt species and, therefore, can be used for solving current problems in the taxonomy and biogeography of this family.     

Population genetic structure of northern Dolly Varden char <Emphasis Type="Italic">Salvelinus malma malma</Emphasis> in Asia and North America

The level of genetic differentiation of northern Dolly Varden char Salvelinus malma malma from Asia and North America was evaluated using the data on mtDNA variation (regions ND1/ND2, ND5/ND6, and Cytb/D-loop) obtained by means of PCR-RFLP analysis. For S. m. malma, the mean values of haplo-type and nucleotide diversity were 0.5261 ± 0.00388 and 0.001558, respectively. The mean estimate of the population nucleotide divergence constituted 0.055%. It was demonstrated that S. m. malma on the most part of the species range examined (drainages of the Beaufort Sea, Chukotka Sea, Bering Sea, and the Sea of Okhotsk) was characterized by the population genetic structure with the low level of genetic differentiation and divergence. At the same time, populations from the Pacific Ocean Gulf of Alaska demonstrated marked genetic differentiation, supported by the high pairwise G4ST values (from 0.4198 to 0.5211) and nucleotide divergence estimates (mean divergence, 0.129%), from Asian and North American populations. Analysis of molecular variance (AMOVA) showed that most of the mtDNA variation in S. m. malma fell in the intrapopulation component (72.5%). At the same time, the differences between the populations (21.1%) and between the regions (6.4%) made lower contribution to the total variation.     

Molecular characterization of clonal population structure and biogeography of arctic apomictic Daphnia from Greenland and Iceland

As part of a continuing international project to characterize the population genetic (clonal) structure of arctic members of the Daphnia pulex complex, 34 populations from western Iceland (N= 1373), and 76 populations from western Greenland (N= 2917), were surveyed for allozymic variation at six polymorphic enzyme loci. Mean clonal richness (±SE) was 1.91±0.19 and 1.50±0.12 for Iceland and Greenland populations, respectively. Mean clonal diversity (±1 SE) was 1.20±0.07 and 1.13±0.04 for Iceland and Greenland populations, respectively. Four widely distributed clones constituted 92.2% of the total animals surveyed from Iceland, while three locally abundant clones constituted 80.1% of the total animals collected primarily from Disko Island, western Greenland. Selected populations were screened for mitochondrial (mt)DNA variation using PCR-based RFLP analysis of a 2100 bp fragment containing part of the ND4 and ND5 genes. One mtDNA haplotype was very widespread in both western Greenland and western Iceland, although, a number of mutational derivatives were also detected. These data indicate the potential for long-distance dispersal of mtDNA lineages, of the order of hundreds or thousands of kilometers across the arctic. Phylogenetic analysis of the sequence of a 254 base pair (bp) fragment of the control region of the mtDNA molecule revealed two major clades one of which consisted solely of non-melanic lineages, and the other of which consisted almost exclusively of melanic lineages (i.e. one non-melanic lineage also clustered in this clade). Sequence divergence between the two clades averaged 7.3%. Both mitochondrial analyses did not reveal any distinct intraregional clustering of lineages. We discuss our results in reference to previous molecular work done on this arctic Daphnia complex, and we attempt to infer phylogeographic patterning based on geological/glaciological historical events in this region of the arctic.     

Mitochondrial DNA variation in Pacific capelin (<Emphasis Type="Italic">Mallotus villosus catervarius</Emphasis>) from the Sea of Okhotsk,inferred from PCR-RFLP analysis

Four population samples of Pacific capelin Mallotus villosus catervarius (Pennat, 1784) from geographically distant localities in the Sea of Okhotsk, Tauy Bay and the eastern coast of the Sakhalin Island, were examined using PCR-RFLP analysis of three mitochondrial DNA regions (A8/A6/COIII/ND3, ND3/ND4L/ND4, and ND5/ND6). The nucleotide divergence of mtDNA sequences among the samples, as well as the analysis of geographic heterogeneity of the haplotype frequencies and quantitative estimation of genetic differentiation performed by means of AMOVA, showed that the samples examined belonged to one panmictic population. Genealogic analysis of the mtDNA variation structure was carried out. It was demonstrated that the high level of haplotype diversity (0.9639 ± 0.00015) along with the low level of nucleotide diversity (0.003818 ± 0.0000003) pointed to the exponential rate of population growth of the capelin from the Sea of Okhotsk, which rather recently in its evolutionary history faced the bottleneck effect.     

The genetic structure of endangered indigenous populations of the amago salmon, <Emphasis Type="Italic">Oncorhynchus</Emphasis><Emphasis Type="Italic">masou</Emphasis><Emphasis Type="Italic">ishikawae</Emphasis>, in Japan

The amago salmon, Oncorhynchus masou ishikawae, is an endemic subspecies of O. masou in Japan. Owing to the extensive stocking of hatchery fish throughout Japan, indigenous populations of O. m. ishikawae are now on the verge of extinction. We examined the genetic effects of stocking hatchery fish on wild populations in the River Koza, Japan, using microsatellite and mitochondrial DNA (mtDNA) markers. For mtDNA, haplotype mt1, which is common in wild populations, was present exclusively in isolated wild populations assumed to be unaffected by previous stocking, while it was never observed in hatchery fish. Genetic diversity was much higher in wild populations in the stocked area, which shared many mtDNA haplotypes with hatchery fish, than in isolated wild populations with haplotype mt1. Pairwise F _ST estimates based on microsatellites showed significant differentiation among the isolated populations with many microsatellite loci monomorphic. Significant deviation from Hardy–Weinberg equilibrium was observed in wild populations in the area subject to stocking, where a Bayesian-based assignment test showed a high level of introgression with hatchery fish. These results suggest that wild populations with haplotype mt1, which became isolated through anthropogenic environmental change in the 1950–1960s, represent indigenous populations of O. m. ishikawae in the River Koza. They have low genetic diversity, most likely caused by genetic bottlenecks following damming and environmental deterioration, while stocking of hatchery fish over the past 30 years apparently had a large impact on the genetic structure of wild populations in the main channel of the River Koza.     

Resolving taxonomic uncertainties using molecular systematics: <Emphasis Type="Italic">Salmo dentex</Emphasis> and the Balkan trout community

The genetic structure of Salmo dentex and its phylogenetic relations to sympatric salmonids in the Neretva and Skadar River basins were evaluated using mitochondrial DNA (mtDNA) control region, eight microsatellites, and somatolactin (SL) gene. In the Neretva River basin of Bosnia–Herzegovina, the results based on mtDNA analysis showed extensive haplotype sharing between S. marmoratus, S. dentex, and S. trutta, and were therefore not conclusive; however, F-statistics and assignment testing based on nuclear DNA markers indicated that S. dentex of the Neretva basin were grouped in a genetically unified cluster with S. marmoratus in the Neretva basin. Using the same analytical approach, S. dentex from the Skadar basin in Montenegro appeared to be genetically distinct from S. marmoratus in the same basin and indistinct from local S. trutta. Molecular data also indicated that S. dentex of the Neretva basin in Bosnia–Herzegovina are not closely related to S. dentex of the Skadar basin in Montenegro. Based on these results, we hypothesize S. dentex to be a particular life history form of S. marmoratus in the Neretva basin and of S. trutta in the Skadar basin. These results clearly demonstrate that S. dentex does not represent a monophyletic lineage and should not be considered a distinct species.     

Spatial and temporal variation in invertebrate consumer diets in forested and herbaceous wetlands

Lake Kronotskoye (the Kronotsky Biosphere State Reserve, south-eastern Kamchatka Peninsula) contains three closely related resident morphotypes charrs, which are considered to be either independent species (white charr Salvelinus albus, longhead charr Salvelinus kronocius, Schmidt’s charr Salvelinus schmidti) or a united lacustrine-riverine charrs, represented by several phenotypes. Salvelinus malma malma is isolated from the lake charr populations by an upstream migration barrier in the Kronotskaya River, which flows out of the lake. We examined the level of genetic variability and divergence of mtDNA both between charrs from Kronotsky Lake and between resident lacustrine charrs and the hypothetical ancestor species S. m. malma. The RFLP-PCR analysis was used to examine six regions (ND1/ND2, ND3/ND4L/ND4, ND5/ND6, COI/COII/A8, A8/A6/COIII/ND3, and Cytb/D-loop), comprising ~80% of the mtDNA. Significantly different levels of diversity were found among the populations of lacustrine charrs. S. albus and S. schmidti had the highest indices of mtDNA diversity among the investigated populations from the different habitats. Heterogeneity tests revealed highly significant differentiation among lake populations and among riverine (Kronotskaya River) and lake (Lake Kronotskoye) populations of charrs, indicating their reproductive isolation. Hierarchical analysis of molecular variance revealed the following regularities of diversity distribution: the high proportion of interpopulation variation (93.25%) and low but statistically significant subdivision between charr populations (6.75%, P < 0.001). Results of the present study suggest that the populations of S. albus, S. kronocius, S. schmidti belong to the S. m. malma phylogenetic group. The divergence value of mtDNA of resident charrs (0.0357–0.0010%) does not exceed the intraspecific variability of S. m. malma. The analysis of the mtDNA haplotypes genealogy of charrs showed that the low values of nucleotide divergence reflect a short period from the beginning of divergence from the ancestral lineages and are due to ancestral polymorphism, as well as to haplotype exchange between the diverged phylogenetic groups as a result of introgressive hybridization.     

Morphology and mitochondrial DNA variation of the Siberian whitefish <Emphasis Type="Italic">Coregonus lavaretus pidschian</Emphasis> (Gmelin) in the upstream water bodies of the Ob and Yenisei Rivers

Combining morphological, ecological and genetic analyses, we compared patterns of diversification within and among populations of the southern Siberian whitefish species Coregonus lavaretus pidschian (Gmelin) to illuminate their evolutionary history. Using sequencing data from 1,930 bp of NADH dehydrogenase subunit 1 (ND1) mitochondrial DNA regions, we documented phylogeographic relationships among populations and developed a phylogeny of mtDNA haplotypes. We found significant differences in the perforated lateral-line scale numbers within and between some populations. Clear differences in the number of gill rakers on the first branchial arch were only exhibited between populations of C. l. pidschian and Coregonus lavaretus pravdinellus Dulkeit. Concordance between different morphological groups based on two meristic traits and mtDNA patterns was also tested.     

Differentiation of the Dolly Varden Char (Salvelinus malma) and the Taranetz Char (Salvelinus taranetzi) Inferred from the PCR–RFLP Analysis of Mitochondrial DNA

Genetic differentiation of two sympatric char species, the Dolly Varden char (Salvelinus malma Walbaum) and the Taranetz char (S. taranetzi Kaganovski), has been studied. Restriction fragment length polymorphism (RFLP) analysis has been used to compare three mitochondrial DNA (mtDNA) fragments (ND1/ND2, ND5/ND6, and Cytb/D-loop) amplified via polymerase chain reaction (PCR). The divergence between S. malma and S. taranetzi inferred from mtDNA nucleotide sequences is 2.8%; between S. leucomaenis and S. taranetzi, 7.1%; and between S. malma and S. leucomaenis, 7.5%. The absence of common haplotypes and the degree of divergence indicate that the Dolly Varden char and the Taranetz char are genetically isolated and confirm that S. taranetzimay be regarded as a separate species.     

The genetic population structure of lacustrine sockeye salmon, <Emphasis Type="Italic">Oncorhynchus nerka,</Emphasis> in Japan as the endangered species

Lacustrine sockeye salmon (Oncorhynchus nerka) are listed as an endangered species in Japan despite little genetic information on their population structure. In order to clarify the genetic diversity and structure of Japanese populations for evaluating on the bottleneck effect and an endangered species, we analyzed the ND5 region of mitochondrial DNA (mtDNA) and 45 single nucleotide polymorphisms (SNPs) in 640 lacustrine sockeye salmon in Japan and 80 anadromous sockeye salmon in Iliamna Lake of Alaska. The genetic diversity of the Japanese population in both mtDNA and SNPs was significantly less than that of the Iliamna Lake population. Moreover, all Japanese populations had SNP loci deviating from the HWE. In spite of low genetic diversity, the SNP analyses resulted that the Japanese population was significantly divided into three groups. These suggest that Japanese sockeye salmon populations should be protected as an endangered species and genetically disturbed by the hatchery program and transplantations.     

mtDNA indicates profound population structure in Indian tiger (Panthera tigris tigris)

We analyzed mtDNA polymorphisms (parts of control region, ND5, ND2, Cytb, 12S, together 902 bp) in 59 scat and 18 tissue samples from 13 Indian populations of the critically endangered Indian tiger (Panthera tigris tigris), along with zoo animals as reference. Northern tiger populations exhibit two unique haplotypes suggesting genetic isolation. Western populations from Sariska (extinct in 2004) and Ranthambore are genetically similar, such that Ranthambore could serve as a source for reintroduction in Sariska. Zoo populations maintain mitochondrial lineages that are rare or absent in the wild.     

Reconciling molecular signatures across markers: mitochondrial DNA confirms founder effect in invasive North American house finches (<Emphasis Type="Italic">Carpodacus mexicanus</Emphasis>)

Well-characterized species introductions provide opportunities to compare the genetic signatures of known founder effects across classes of molecular markers. The release of small numbers of house finches (Carpodacus mexicanus) into the eastern United States in the 1940s led to substantial interest in the effects of this introduction on genetic diversity in this now abundant species, an issue that has been highlighted by a recent Mycoplasma disease epidemic that most intensively affects the introduced and potentially genetically depauperate house finch populations. Previous studies comparing genetic diversity levels in native and introduced house finch populations produced seemingly disparate results: comparisons based on amplified fragment length polymorphism, RFLP mtDNA, and allozyme markers found essentially equivalent levels of diversity in eastern and western populations, whereas microsatellite markers showed clear reductions in diversity in the introduced populations. Here we employ sequence variation at the ND2 mtDNA locus to further compare levels of diversity between the four native and five introduced house finch populations that were previously examined in the microsatellite study. We found substantially lower ND2 haplotype richness and diversity across all introduced populations of house finches. The majority of sequence variation (78%) was detected within subpopulations, with the remainder (22%) explained by the historical status of each population (native or introduced). Our results are consistent with previous microsatellite evidence for a founder effect during the introduction of eastern house finches, and suggest that the mtDNA founder effect was particularly severe, likely owing to a male-biased sex ratio at the time of introduction coupled with the lower effective population size of clonally inherited markers. We discuss how the inconsistencies between past studies of house finch diversity can inform the usefulness of distinct marker sets for detecting molecular signatures of founder events.     

Genetic population structure of a reef goby, <Emphasis Type="Italic">Bathygobius cocosensis</Emphasis>, in the northwestern Pacific

To investigate the genetic population structure of a reef goby, Bathygobius cocosensis, in the northwestern Pacific in relation to major oceanic currents (North Equatorial and Kuroshio Currents), the mitochondrial DNA (mtDNA) phylogeny and demography of the species were inferred from partial nucleotide sequences of the mitochondrial NADH dehydrogenase 5 subunit (ND5) gene (945 bp). The resultant mtDNA tree and population genetic analyses showed that the Bonin Islands’ population apparently differed from the other populations in the Japan–Ryukyu–Guam region, which lie along the routes of the North Equatorial and Kuroshio Currents. A test calculation indicated the divergence date to be approximately 1 million years ago. The population expansion parameters indicated that the Bonin Islands’ population had survived during the fluctuations in sea level in the Pleistocene–Holocene age and that the population had expanded thereafter.     

Mitochondrial DNA variation and population genetic structure of the northern redbelly dace (Phoxinus eos)

Two sections of the control region and the genes coding for NADH dehydrogenase sub-units 5 and 6 (ND-5/6) of mitochondrial DNA (mtDNA) were amplified from Phoxinus eos with the polymerase chain reaction. Both sections of the control region were sequenced directly while the ND-5/6 fragment was sequenced in from each end only. Additionally, the entire ND-5/6 fragment was examined for sequence variation using RFLP analysis. No sequence variation was detected in the control region among 70 individuals sampled from 18 populations across three Ontario regions (Spanish River, Madawaska R. and Cataraqui R.). To examine ND-5/6 variation, a total of 75 individuals were sampled from five populations representing two of the three regions (Madawaska River and Cataraqui R.). Six haplotypes were detected by direct sequencing and four by RFLP analysis. Estimates of population subdivision from RFLP data, sequence analysis, and the two data sets combined for the ND-5/6 fragment, suggest that gene flow is restricted within and between regions. However, estimates of sequence divergence for both sequence and RFLP analysis of this fragment suggested that populations were either founded by already differentiated populations or that populations were founded by a single stock and divergence between regions occurred prior to isolation of populations within regions. These estimates of population structure are much greater than those obtained from allozyme analysis. Additionally, high levels of heterozygosity in nuclear DNA, but low mtDNA diversity suggests that populations have experienced reductions in population size sufficient to reduce only mtDNA variation. Random lineage extinction and limited time for the accumulation of new mutations are likely responsible for low levels of mtDNA variation in ND-5/6 and the control region, while functional constraints may limit variation more than expected in the control region in dace and other fishes.     

Genetic Differentiation of Three Sympatric Charr Species from the Genus SalvelinusInferred from PCR–RFLP Analysis of Mitochondrial DNA

We examined differentiation of three sympatric charr species Dolly Varden charr Salvelinus malmaWalbaum, white-spotted charrS.leucomaenisPallas, and Levanidov charrS. levanidovi Chereshnev, Scopetz, Gudkov. Using restriction fragment length polymorphism analysis (RFLP), three mitochondrial DNA fragments (ND1/ND2, ND5/ND6, and Cytb/D-loop) amplified in PCR were compared. The divergence of mtDNA nucleotide sequences betweenS. levanidovi and S. leucomaenis was about 9.7%, between S. levanidovi and S. malma, 8.7%, and between S. malma andS. laecomaenis, 7.5%. These results indicate genetic isolation of the species examined and support the earlier suggestion on closeness of Levanidov charr to the common ancestor of the genus Salvelinus.     

Nucleotide diversity of a ND5 fragment confirms that population expansion is the most suitable explanation for the mtDNA haplotype polymorphism of <Emphasis Type="Italic">Drosophila subobscura</Emphasis>

Results from mitochondria (mt) DNA restriction site analyses (RSAs) have revealed that wild populations of Drosophila subobscura are formed by two common (I and II) and some rare, often endemic, low-frequency haplotypes. In the study reported here, we analysed nucleotide diversity in a 942-bp fragment of the mtDNA ND5 gene in 48 D. subobscura individuals captured from three populations that showed haplotypes I, II or the less common ones, as well as in one additional individual belonging to D. guanche that was taken as an outgroup. RSAs and sequencing results were compared. The two approaches yielded similar nucleotide variability parameters, suggesting a consistency in the results obtained from mtDNA dynamics in natural populations of D. subobscura. Patterns of polymorphism at ND5 are most consistent with the hypothesis of population expansion after a bottleneck that may have occurred since the last glaciation or which may occur seasonally after the summer and winter. However, we cannot rule out that selection has a role in maintaining the two major haplotypes at intermediate frequencies in worldwide populations of D. subobscura.     

PCR–RFLP analysis of mitochondrial DNA in tench Tinca tinca

Polymorphism was detected at ND1, ND6, D‐loop and cyt b segments of mtDNA in 105 tench (Tinca tinca L.), using the polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) technique and five composite haplotypes were identified. The diversity indices and the results of the population comparisons revealed that the identified markers provide a powerful tool for further studies on this species.     

Phylogeography of northern Dolly Varden Salvelinus malma malma based on analysis of mitochondrial DNA

The northern Dolly Varden, Salvelinus malma malma, is a typical representative of arctic fauna distributed in northeastern Asia and northwestern North America. Because its spawning habitats were affected by Pleistocene glacial advances over most of its natural range, S. m. malma is among the most interesting objects of phylogeographic and microevolutionary studies. We reconstructed the genealogy of mtDNA haplotypes from 27 Alaskan and Asian populations to study the influence of glacial and geological vicariance events on the contemporary population genetic structure, phylogeographic subdivision and distribution of the northern Dolly Varden. Analysis of restriction site states in three PCR‐amplified mtDNA regions (ND1/ND2, ND5/ND6, Cytb/D‐loop; 47% of the mitochondrial genome) resolved 75 haplotypes in 436 fish. Similar patterns of subspecific variation apparent from hierarchical diversity and nested clade analyses of mtDNA haplotypes identify weak spatial differentiation and low levels of divergence. Our results suggest that (1) demographic history has been influenced by historical range expansions and recent isolation by distance, (2) present populations from Asia and North America were colonized from one main Beringian Refugium, and (3) that this taxon's ancestral population probably experienced a bottleneck in the Beringian Refugium during the late Pleistocene (Wisconsin) glacial period.     

Phylogeographic differentiation in the mitochondrial control region in the koala, Phascolarctos cinereus (Goldfuss 1817)

The koala, Phascolarctos cinereus, is a geographically widespread species endemic to Australia, with three currently recognized subspecies: P.c. adustus, P.c. cinereus, and P.c. victor. Intraspecific variation in the mitochondrial DNA (mtDNA) control region was examined in over 200 animals from 16 representative populations throughout the species’ range. Eighteen different haplotypes were defined in the ≈ 860 bp mtDNA control region, as determined by heteroduplex analysis/temperature gradient gel electrophoresis (HDA/TGGE). Any single population typically possessed only one or two haplotypes yielding an average within-population haplotypic diversity of 0.180 ± 0.003, and nucleotide diversity of 0.16%. Overall, mtDNA control region sequence diversity between populations averaged 0.67%, and ranged from 0% to 1.56%. Nucleotide divergence between populations averaged 0.51%, and ranged from 0% to 1.53%. Neighbour-joining methods revealed limited phylogenetic distinction between geographically distant populations of koalas, and tentative support for a single evolutionarily significant unit (ESU). This is consistent with previous suggestions that the morphological differences formalized by subspecific taxonomy may be interpreted as clinal variation. Significant differentiation in mtDNA-haplotype frequencies between localities suggested that little gene flow currently exists among populations. When combined with microsatellite analysis, which has revealed substantial differentiation among koala populations, we conclude that the appropriate short-term management unit (MU) for koalas is the local population.