Change in nucleotide sequence of mitochondrial DNA from cytochrome b gene in chars of Salvelinus genus

Nucleotide sequences of two (405- and 1050-bp) regions of mitochondrial DNA (mtDNA) cytochrome c gene were established in chars of the genus Salvelinus from Russian Far East and Siberia. Based on the divergence and phylogenetic analysis of nucleotide sequences of the mtDNA cytochrome c gene, S. laecomaenis was shown to carry the most ancient mitochondrial lineage, which is close to the ancestral one. The archaic mtDNA of S. levanidovi occupied an isolated position on the phylogenetic trees. The mtDNA lineage of the southern S. malma was close to the S. alpinus-S. malma complex group. Within the S. alpinus-S. malma complex, three groups of mtDNA types having particular geographic distributions were distinguished. The Kolyma-Chukotka group includes lake S. taranetzi, S. boganidae and S. elgyticus from Chukotka, lake chars from Kolyma. The Okhotsk group is represented by northern S. malma, lake chars from northern Sea of Okhotsk, and anadromous S. taranetzi. The Siberian group is close to the Okhotsk one and consists of Taimyr and Baikal region chars as well as Arctic char from Finland. The divergence of char mitochondrial lineages was dated to the Pliocene-Pleistocene.     

Variability of nucleotide sequences of the mitochondrial DNA cytochrome c gene in dolly varden and taranetz char

Nucleotide sequence of the 307-bp fragment of the mitochondrial DNA cytochrome b gene was determined in representatives of the three species of the Salvelinus genus, specifically, dolly varden char (S. malma), taranetz char (S. taranetzi), and white-spotted char (S. leucomaenis). These results pointed to a high level of mitochondrial DNA (mtDNA) divergence between white-spotted char and dolly varden char, on the one hand, and taranetz char, on the other (the mean d value was 5.45%). However, the divergence between the dolly varden char and taranetz char was only 0.81%, which is comparable with the level of intraspecific divergence in the dolly varden char (d = 0.87%). It was shown that the dolly varden char mitochondrial gene pool contained DNA lineages differing from the main mtDNA pool at least in the taranetz char-specific mitochondrial lineages. One of these dolly varden char mtDNA lineages was characterized by the presence of the restriction endonuclease MspI-D variant of the cytochrome b gene. This lineage was widely distributed in the Chukotka populations but it was not detected in the Yana River (Okhotsk sea) populations. These findings suggest that dolly varden char has a more ancient evolutionary lineage, diverging from the common ancestor earlier than did taranetz char.     

Variation of cytochrome b gene in sympatric chars from kronotsky lake (Kamchatka Peninsula)

In previously identified sympatric char forms (species) from Kronotsky Lake (Kamchatka Peninsula) and a number of other chars from the genus Salvelinus, nucleotide sequences of the mtDNA cytochrome b gene were determined and phylogenetic analysis of these chars was performed. The divergence estimate of the cytochrome b gene in the Kronotsky char group (0.48%) coincided with the interpopulation difference estimates in northern Dolly Varden, which indicates a relatively recent divergence of the chars from Kronotsky Lake. Haplotypes of each form of the Kronotsky Lake chars are divided into two groups, which belonged to two different phylogenetic mtDNA lineages of northern Dolly Varden from Chukotka and Kamchatka. The formation of the Kronotsky char forms proved to be unrelated to the observed variability of the mtDNA cytochrome b gene.     

Genetic divergence of mitochondrial DNA in white char <Emphasis Type="Italic">Salvelinus albus</Emphasis> and northern Dolly Varden char <Emphasis Type="Italic">Salvelinus malma malma</Emphasis>

Comparative analysis of mitochondrial DNA variation was performed in white char Salvelinus albus Glubokovsky and in its putative ancestor species, northern Dolly Varden char Salvelinus malma malma Walbaum. Highly statistically significant differentiation of S. albus and S. m. malma in the areas of sympatric (Kamchatka River basin) and allopatric (Kronotskoe Lake and Kronotskaya River) residence was demonstrated. The mtDNA divergence between S. albus and S. m. malma did not exceed the range of intraspecific variation in the populations of northern Dolly Varden char. At the same time, clusterization pattern of the Salvelinus chars provides hypothesis on the common origin of two allopatric populations of white char. Genealogical analysis of haplotypes indicates that S. albus and S. m. malma currently demonstrate incomplete divergence of mitochondrial lineages. The low nucleotide divergence estimates between S. albus and S. m. malma reflect the short time period since the beginning of the divergence of ancestral lineages. These estimates are determined by ancestral polymorphism and haplotype exchange between the diverged phylogenetic groups as a result of introgressive hybridization.     

Comparative restriction enzyme analysis of mitochondrial DNA in several populations of lake resident chars of the genus Salvelinus

Restriction enzyme analysis was employed in studying the mitochondrial DNA (mtDNA) ATPase6/ND4L region in several Northeast Asian populations of resident lake char of the genus Salvelinus. On evidence of mitotypes, genetic similarity was assumed for populations of neiva (Ueginskoe Lake), lake resident char from the northern coast of the Sea of Okhotsk (Mak-Mak Lake and Elekchanskie lakes), and dolly varden. Mitotype AAAA proved to be common for these populations. Lake char populations of the Juliet and Maxi lakes (the basin of the Kolyma river) had mitotype DBAA, which is similar to mitotype DBAB observed earlier in Taranetz char S. taranetzi from Chukotka. The divergence between mitotypes AAA and DBAA was estimated at 0.3%. Different origins were assumed for the lake resident char populations from the basins of the Sea of Okhotsk and of the Kolyma River, the former originating in the Pacific and the latter, in the Arctic basins.     

Genetic Differentiation Inferred from Data on Restriction Analysis of Mitochondrial DNA in the Northern and Southern Forms of the Dolly Varden Char

Restriction fragment length polymorphism of the ATPase6/ND4L and cytochrome b regions of mitochondrial DNA (mtDNA) was studied in populations of the northern and southern forms of the Dolly Varden char Salvelinus malma from the Asian coast of the northern part of the Pacific Ocean. Seven genotypes of mtDNA were found, of which AAAA/ACCC and BBAC/ACEC characterize the northern and southern groups of populations, respectively. By the identity test, the studied populations of the Chukotka and Kamchatka peninsulas and the northern coast of the Sea of Okhotsk significantly differed from the studied populations of Sakhalin Island, Primorye, and Kuril Islands.     

Differentiation of Dolly Varden Char <Emphasis Type="Italic">Salvelinus malma</Emphasis> from Asia and North America Inferred from PCR-RFLP Analysis of Mitochondrial DNA

Genetic differentiation of Dolly Varden char Salvelinus malma Walbaum from the Asian and North American Pacific coasts was studied. We examined restriction fragment length polymorphism of three mitochondrial DNA (mtDNA) fragments amplified in polymerase chain reaction, which encoded four NADH dehydrogenase subunits, the cytochrome b gene, and a D-loop segment. The mtDNA haplotypes were shown to form three phylogenetic groups, whose geographic distribution corresponded to three Dolly Varden subspecies: S. malma malma, S. malma krascheninnikovi, and S. malma lordi. The nucleotide sequence divergence between S. malma malma and S. malma krascheninnikovi was 3.8%; between S. malma malma and S. malma lordi, 3.1%; and between S. malma krascheninnikovi and S. malma lordi, 2.5%. The northern Dolly Varden S. malma malma from Asia was shown to be genetically identical to that from North America.__________Translated from Genetika, Vol. 41, No. 5, 2005, pp. 626–634.Original Russian Text Copyright © 2005 by Oleinik, Skurikhina, Brykov, Crane, Wenburg.     

Introgressive hybridization of chars of the genus <Emphasis Type="Italic">Salvelinus</Emphasis> as inferred from mitochondrial DNA variation

Nucleotide sequence analysis of the mitochondrial cytochrome b gene in the genus Salvelinus revealed hybrids with S. leucomaenis in S. malma malma populations of the Northern Okhotsk Sea basin. Hybrids of S. m. malma and S. m. krascheninnikovi were found in S. m. malma populations from the Northern Okhotsk Sea basin and Kamchatka. The findings testify to a secondary contact and mtDNA transfer between these species and forms. It was assumed that introgressive hybridization took place both long ago and relatively recently and occurred in one direction: from S. leucomaenis or S. m. krascheninnikovi to S. m. malma.Translated from Genetika, Vol. 40, No. 12, 2004, pp. 1678–1685.Original Russian Text Copyright © 2004 by Radchenko.     

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.     

Comparative Restriction Enzyme Analysis of Mitochondrial DNA in Several Populations of Lake Resident Chars of the Genus Salvelinus

Restriction enzyme analysis was employed in studying the mitochondrial DNA (mtDNA) ATPase6/ND4L region in several Northeast Asian populations of resident lake char of the genus Salvelinus. On evidence of mitotypes, genetic similarity was assumed for populations of neiva (Ueginskoe Lake), lake resident char from the northern coast of the Sea of Okhotsk (Mak-Mak Lake and Elekchanskie lakes), and dolly varden. Mitotype AAAA proved to be common for these populations. Lake char populations of the Juliet and Maxi lakes (the basin of the Kolyma River) had mitotype DBAA, which is similar to mitotype DBAB observed earlier in Taranetz char from Chukotka. The divergence between mitotypes AAAA and DBAA was estimated at 0.3%. Different origins were assumed for the lake resident char populations from the basins of the Sea of Okhotsk and of the Kolyma River, the former originating in the Pacific and the latter, in the Arctic basins.     

Variation of cytochrome b gene in sympatric chars from Kronotsky Lake (Kamchatka oblast)

In previously identified sympatric char forms (species) from Kronotsky Lake (Kamchatka Peninsula) and a number of other chars from the genus Salvelinus, nucleotide sequences of the mtDNA cytochrome b gene were determined and phylogenetic analysis of these forms was performed. The divergence of the cytochrome b gene in the Kronotsky char group (0.48%) coincided with the interpopulation difference estimates in northern Dolly Varden, which indicates a relatively recent divergence of the chars from Kronotsky Lake. Haplotypes of each form of the Kronotsky Lake chars are divided into two groups, which belonged to two different phylogenetic mtDNA lineages of northern Dolly Varden from Chukotka and Kamchatka. The formation of the Kronotsky char forms proved to be unrelated to the observed variability of the mtDNA cytochrome b gene.     

Problems of molecular systematics of the genus Salvelinus based on nucleotide sequence variability of the major noncoding region of mitochondrial DNA

Variability of the major noncoding mitochondrial DNA region of char (Salvelinus alpinus complex) was analyzed by the median network method on the basis of the literature data on nucleotide sequences of this region. A significant portion (21%) of the polymorphic sites of char mtDNA was shown to be evolutionarily unstable, which results in mutation homoplasia at these sites. Using median-joining analysis the chars were divided into three phylogeographical groups: Arctic, Beringian, and chars occurring in the vast region from Lake Baikal and Taimyr Peninsula to the Atlantic Ocean. The latter group includes three subgroups (Siberian, Atlantic, and Acadian), which diverged from a common ancestor. In the Arctic group, the branch of Taranetz char S. taranetzi is clearly separated. It diverged from the common ancestor earlier than S. boganidae and S. elgyticus occurring on the Chukotka Peninsula species and S. a. erythrinus occurring in Alaska. The Beringian group, which includes dolly varden char from various regions of Asia and America, is an intermediate between the two char groups. The main problems of char systematics inferred from the major noncoding mtDNA region variability arise from the high level of mutation homoplasia.     

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.     

Genetic Divergence of Chars of the Genus <Emphasis Type="Italic">Salvelinus</Emphasis> from Kronotsky Lake (Kamchatka Peninsula)

Chars of the genus Salvelinus, inhabiting lakes and lake-river systems, comprise morphological and ecological forms whose taxonomic status is under dispute. In the present work, we have examined genetic variation and divergence in various chars from the Kronotsky lake basin: the lacustrine chars (white, nose, and longhead) and riverine Dolly Varden Salvelinus malma. The study was conducted using analysis of allozyme and microsatellite loci, myogens, RAPD, and restriction analysis of two mtDNA segments. The estimates of heterozygosity at allozyme and microsatellite loci were similar to the corresponding parameters in populations of northern Dolly Varden and Arctic char. Heterozygote deficit was recorded in both samples of separate forms, and in the total sample of all chars from Kronotsky Lake. For allozyme and microsatellite loci, appreciable genetic differentiation among the samples of different char forms was found, which was comparable to that among the spatially isolated populations of northern Dolly Varden. This result indicates reproductive isolation among the char forms examined. However, this isolation is not complete, because no fixed differences between the forms by any of the genetic systems analyzed were found. The genetic differentiation among different forms of lacustrine chars, which corresponds to the interpopulation rather than interspecific level, is thought to be explained by their comparatively recent divergence.__________Translated from Genetika, Vol. 41, No. 8, 2005, pp. 1096–1107.Original Russian Text Copyright © 2005 by Salmenkova, Omel’chenko, Radchenko, Gordeeva, Rubtsova, Romanov.     

Karyological Differentiation of Northern Dolly Varden and Sympatric Chars of the Genus <Emphasis Type="Italic">Salvelinus</Emphasis> in Northeastern Russia

In many cases the taxonomic status of sympatric chars (with exception of white-spotted char Salvelinus leucomaenis) is not clear and is actively debated. We karyotyped three pairs of sympatric chars inhabiting the Russian Far East-northern Dolly Varden, S. malma malma, and Lavanidov's char, S. levanidovi, from the Yama River (northern coast of the Sea of Okhotsk, Magadan Region), northern Dolly Varden and Taranetz's char, S. taranetzi, from Lake Achchen (east Chukotka), and northern Dolly Varden and white char, S. albus, from the Kamchatka River (Kamchatka Peninsula). Three of them had similar chromosome numbers. But all chars studied had an individual and discrete set of karyotypic characters, which enabled reliable identification each of them by chromosome number, chromosome arm number, and number and location of active nuclear organizer regions.     

Problems of Molecular Systematics of the Genus Salvelinus Based on Nucleotide Sequence Variability in the Major Noncoding Region of Mitochondrial DNA

Variability of the major noncoding mitochondrial DNA region of char (Salvelinus alpinus complex) was analyzed by the median network method on the basis of the literature data on nucleotide sequences of this region. A significant portion (21%) of the polymorphic sites of char mtDNA was shown to be evolutionarily unstable, which results in mutation homoplasia at these sites. Using median-joining analysis the chars were divided into three phylogeographical groups: Arctic, Beringian, and chars occurring in the vast region from Lake Baikal and Taimyr Peninsula to the Atlantic Ocean. The latter group includes three subgroups (Siberian, Atlantic, and Acadian), which diverged from a common ancestor. In the Arctic group, the branch of Taranetz char S. taranetzi is clearly separated. It diverged from the common ancestor earlier than S. boganidae andS. elgyticus occurring on the Chukotka Peninsula species and S. a. erythrinus occurring in Alaska. The Beringian group, which includes dolly varden char from various regions of Asia and America, is an intermediate between the two char groups. The main problems of char systematics inferred from the major noncoding mtDNA region variability arise from the high level of mutation homoplasia.     

The Divergence of the Dolly Varden Char Salvelinus malma in the Asian Northern Pacific Populations Inferred from the PCR–RFLP Analysis of Mitochondrial DNA

Genetic differentiation of the Dolly Varden char Salvelinus malmaWalbaum was studied in five populations from the western part of the Northern Pacific. Using restriction analysis (RFLP), we examined polymorphism of three mitochondrial DNA (mtDNA) fragments amplified in polymerase chain reaction (PCR). MtDNA haplotypes were shown to fall into two phylogenetic groups, which probably reflect the existence of two previously described subspecies of Asian Dolly Varden,S. malma malma and S. malma krascheninnikovi. The divergence of mtDNA nucleotide sequences in the Dolly Varden subspecies (about 4%) corresponds to the differences between the valid char species from the genus Salvelinus.     

Karyotype and relationships of the endemic char Salvelinus krogiusae from Dal'nee Lake (Kamchatka)

Resident (lacustrine) chars, inhabiting many lakes of Kamchatka, Chukotka, and continental coast of the Sea of Okhotsk, are of particular interest for understanding the mechanisms of speciation and evolution of Salvelinus chars. Since one of these, the char from Dal'nee Lake (Paratunka River basin, southeastern Kamchatka), is substantially different from lacustrine-riverine and anadromous chars from the Paratunka River basin in several morphological traits, hemoglobin spectrum, ecology, and reproduction, it is considered an isolated species Salvelinus krogiusae. The karyotype of the resident char from Dal'nee Lake was shown to be variable in most individuals, containing 76 to 80 chromosomes in different cells, with NF = 98. This suggests that the variability of the chromosome number in this char form is explained by Robertsonian translocations. In females and males, cells with respectively 2n = 78 and 2n = 77 prevailed. Heteromorphic sex chromosomes were not found in the karyotype of this char species. Nucleolar-organizing regions (NORs) were mostly detected in the telomeric regions of short arms of large submetacentric chromosomes. These features of the Krogius char karyotype demonstrate that this species is isolated from other Kamchatka chars, being similar to Taranetz char S. taranetzi from Chukotka Peninsula, which is in good agreement with morphological data.     

Occurrence of sympatric charr groups,Salvelinus, Salmonidae,in the lakes of Kamchatka: a legacy of the last glaciations

Six postglacial lakes were studied along both sides of the Kamchatka central mountain range, Russia. Pairs of local morphotypes of species of Arctic charrs, Salvelinus spp., have previously been described from the southernmost lakes while the fish fauna of the four northernmost lakes was studied here for the first time. Phenotypic data support the division of Kamchatkan lacustrine charrs into two groups according to the number of gill rakers and pyloric caeca, as well as snout–dorsal and snout–ventral distances (MANOVA, P < 0·001). These groups respectively correspond to phenotypes commonly referred to as Salvelinus malma and Salvelinus taranetzi. To clarify the identity of these groups, D‐loop and cytochrome b (cytb) region sequences were analysed. Haplotype network analysis of mtDNA shows the salmonids inhabiting four lakes on the south and north are phylogenetically close to either Beringian S. malma or to S. taranetzi from the Chukotka and Kolyma River basins (the mean ± s.e. pairwise per cent sequence divergence is 0·006 ± 0·001). Phenotype–genotype discordance suggests that mitochondrial introgression between species has occurred in the two smallest lakes (<0·5 km²) in the central part of the peninsula. Identical haplotypes of D‐loop and cytb regions were found for the populations of S. taranetzi from the most distant southern and northern lakes, indicating all lakes were colonized by both species simultaneously after the last glacial maximum. Salvelinus taranetzi may have colonized the Kamchatka peninsula from one or both of two different source regions: the Arctic Beringia and the northern coast of the Okhotsk Sea.     

Phylogeography of mitochondrial DNA in South Asian Dolly Varden char <Emphasis Type="Italic">Salvelinus curilus</Emphasis> Pallas, 1814 (Salmoniformes,Salmonidae): Mediated gene introgression?

In 41 individuals of South Asian Dolly Varden char Salvelinus curilus, nucleotide sequences of tRNA-Pro gene fragment (27 bp) and mtDNA control region (483–484 bp) were analyzed. The fish were collected in 20 localities covering virtually the whole range of the species: Kuril Islands, Sakhalin Island, and Primorye. In addition, six individuals of three other char species (S. albus, S. malma, and S. leucomaenis), which are closely related to S. curilus and inhabit the Russian Far East, were examined. In all, we detected 12 different variants of mtDNA haplotypes that formed three distinct groups differing in 14–20 nucleotide positions. The first group consisted of six haplotypes found in S. curilus in Kuril Islands, Sakhalin, and Primorye (mtDNA phylogroup OKHOTSKIA). The second group comprised four haplotypes representing the mtDNA phylogroup BERING, which had been described earlier (Brunner et al, 2001); they were found in S. curilus in Kuril Islands and Sakhalin, as well as in S. albus and S. malma in Kamchatka and northern Kurils. The third group included two haplotypes detected in S. leucomaenis. The existence of two mtDNA lineages (OKHOTSKIA and BERING) in S. curilus from Kurils and Sakhalin was explained by hybridization and mtDNA transfer from S. malma to S. curilus. The absence of the BERING haplotypes in S. curilus from Primorye water reservoirs is related to the physical isolation of the Sea of Okhotsk and Sea of Japan basins in past epochs. On the basis of comparing phylogenetic trees, constructed from the data on allozyme and mtDNA variation, we suggest that in this case, a mediated transfer of mtDNA in Alpinoid chars → S. malma → S. curilus chain could occur.