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.     

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.     

Differentiation of Dolly Varden char Salvelinus malma 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.     

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.     

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.     

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.     

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.     

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.     

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.     

Genetic divergence of sympatric charrs of the genus Salvelinus from Nachikinskoe Lake (Kamchatka)

We studied genetic differentiation of two charr species, Dolly Varden Salvelinus malma malma Walbaum and resident lacustrine charr Salvelinus sp., which sympatrically inhabit Nachikinskoe Lake (the Bol'shaya River basin) in southwestern Kamchatka Peninsula. Using restriction analysis (RFLP), three mitochondrial DNA fragments (ND1/ND2, ND5/ND6, and Cytb/D loop) amplified in polymerase chain reaction (PCR) were compared. The divergence of the mtDNA sequences between Salvelinus sp. and S. malma malma was 2.8%; Salvelinus sp. and S. taranetzi, 0.36%; Salvelinus sp. and S. krogiusae, 0.21%; Salvelinus sp. and S. alpinus, 3.0%. These results point to reproductive isolation of charrs in Nachikinskoe Lake and support the earlier suggestion on a close relationship between Salvelinus sp., S. taranetzi, and S. krogiusae.     

Phylogeography of mitochondrial DNA in South Asian Dolly Varden char Salvelinus curilus 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 DNA 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, an indirect transfer of mtDNA in Alpinoid chars--> S. malma-->S. curilus chain could occur.     

Comparative catches and food habits of dolly varden and arctic charrs,Salvelinus malma and S. alpinus,at Karluk,Alaska, in 1939–1941

Synopsis For many years the seriousness of the predations of dolly varden (Salvelinus malma) upon the eggs, alevins and juveniles of Pacific salmons (Oncorhynchus) has been a controversial subject. Over $ 300,000 was spent in Western Alaska from 1920 to 1941 for bounties on dolly varden in the belief that they were serious salmon predators. The writter undertook this study because of an intense desire to evaluate the need and justification for such a bounty system. The incidental examination of stomach contents of charrs which were captured for tagging or for tag recovery purposes from Karluk Lake and Karluk River on Kodiak Island, Alaska, and from Shelikof Strait and Uyak Bay during the summer months of 1939, 1940 and 1941 presented an excellent opportunity to pursue such a project. Stomachs were collected and contents analyzed from 1,992 arctic charr (Salvelinus alpinus) and 2,565 dolly varden charr (Salvelinus malma), taken by beach seine, gill net, fyke net, hook and line and by weir trap from Karluk Lake; from 956 dolly varden taken by seine and weir trap from the Karluk River; and from 462 dolly varden taken by beach seine and commercial salmon traps in Uyak Bay. Among the more than 5,000 charr stomachs examined, only 42 were found to contain salmon smolts, parr or alevin. Examination of 500 stomachs taken from dolly varden charr captured in the lower Karluk River at the height of the red salmon (Oncorhynchus nerka) smolt migration in May of 1939 revealed very little evidence of predation on the smolts in fresh water. There was some evidence that this relationship may have changed after the migrating charr had become adjusted to salt water, although the examination of 460 stomachs of dolly varden captured in salt water revealed nothing particularly incriminating in its role as a salmon predator. The fact that the downstream migration of the dolly varden reached its peak before that of the red salmon smolts was considered significant. Examination of the 4,500 charr stomachs at Karluk Lake in the months from April through September led to the discovery that two distinct species of charrs were present, one, the dolly varden, primarily stream-inhabiting and anadromous, and the other, the arctic charr, primarily lake-inhabiting and nonanadromous. Although red salmon eggs constituted 32% of the total volume of food ingested by both species of charr at Karluk Lake, there was considerable evidence that they were consumed in salvage or scavenger feeding, and there was practically no evidence that they were consumed as a result of predatory feeding. Only five, or one-tenth of one percent, of the thousands of charr stomachs examined at Karluk Lake contained red salmon parr or alevin.A summary of the contrasts in the feeding and migratory habits and physical characteristics of Salvelinus malma and Salvelinus alpinus led to the conclusion that alpinus was a more serious potential salmon predator than malma at Karluk Lake. In view of the fact that alpinus was seldom reached by the predator-control program whereas malma was being systematically destroyed only because it was more easily captured due to its migratory tendencies, it was recommended that the predator control program be terminated in Karluk waters.Published post mortem in shortened and slightly edited version. Reprint requests to the Editor.     

Population genetic structure of the char species of the Northern Kuril Islands and the rank of the Dolly Varden Char in the system of the genus Salvelinus (Salmonidae: Teleostei)

Analysis of the taxonomic position of most species and forms of the char (genus Salvelinus, Salmonidae: Teleostei) was made based on RAPD-PCR. The material was represented by samples from 29 populations from the Kuril Islands, coast of the Sea of Okhotsk, Kamchatka, Chukotka, Taymyr, Transbaikalia, the Kola Peninsula, Svalbard, Finland, and North America. It was shown that the genus Salvelinus splits into three well-justified clusters: (1) all the forms assigned to the Salvelinus alpinus--S. malma complex; (2) two samples of the White-Spotted Char from the Southern Kuril Islands and from Kamchatka; (3) two North American species, S. fontinalis and S. namaycush (samples of the North American species S. confluentis were absent from the collection). Analysis of the absolute values of genetic disctances of the S. alpinus--S. malma forms relative to S. leucomaenis, S. fontinalis, and S. namaycush revealed distances approaching the species rank between the following isolates: Frolikh Char, Mountain Char, Black Lake Char, Goggle-Eyed Char, and Neyva Char. Samples of Dolly Varden currently considered as "S. malma", do not constitute a separate cluster, falling within the group of the Arctic char S. alpinus. This conclusion is supported by the analysis of the results of three series of experiments by R. Phillips on ITS1 ribosome genes (Pleute et al., 1992; Phillips et al., 1995; Phillips et al., 1999). This indicates the infraspecific rank of malma within S. alpinus. Isolated populations of "Salvethymus svetovidovi" from the lake Elgygytgyn (Chukotka Peninsula) and of the char from the lake Chyornoye (Onekotan Island), recently described as S. gritzenkoi (Vasil'eva, Stygar, 2000), fell withing the S. alpinus--S. malma complex, the Onekotan char grouped together with another isolate from the same island. Comparison of genetic distances between the samples showed that the differences between the two isolated of Onekotan and migratory forms of the Kuril Islands are approximately equal, yet the homogeneity of the Chyornoye sample is higher than that of the other samples. The revealed 330-nucleotide diagnostic sequence of the Onekotan lake isolate showed identity of part of the fragment with a section of expressed DNA from the library of EST clones derived from the gills of Salmo salar, this possibly indicates the adaptive character of the evolution.     

Population genetic structure of northern Dolly Varden char Salvelinus malma malma 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 haplotype 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 phi(ST) values (from 0.4198 to 0.5211) and nucleotide divergence estimates (mean divergence, 0.129%), from Asian and North American populations. Nested 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.     

Genetic differentiation of charrs in the Russian north and far east

Genetic differences and relationships within charrs Salvelinus alpinus , northern and southern S. malma, S. taranetzi, S. levanidovi and S. leucomaenis , were studied by examination of 31–35 allozyme loci and PCR-RFLP analysis of mtDNA segment (about 2600 bp) including the cytochrome b gene and D-loop. Data for mtDNA were also obtained for samples of S. drjagini, S. albus and for European samples of S. alpinus. Generally allozyme and mtDNA segment analyses produced similar pictures of genetic divergence and relationships. The southern S. malma exhibited the greatest genetic variability (allozyme heterozygosity and mtDNA haplotype diversity). S. alpinus, S. malma, S. levanidovi and S. leucomaenis had fixed differences from one another at various allozyme loci, and differed in fixed (but polymorphic in S. malma ) individual mtDNA haplotypes produced with MspI restrictase. Such genetic individuality is typical of distinct taxa. S. taranetzi is closest genetically to allopatric S. alpinus , but exhibited fixed differences from the northern S. malma in sympatry. In all samples of S. alpinus the only mtDNA haplotype was shared with S. drjagini. S. albus shared its single haplotype with the northern S. malma from the same locality. S. leucomaenis is differentiated from the other species to the greatest extent, possibly reflecting an early evolutionary divergence. Some evolutionary and taxonomic implications of the genetic relationships of charrs studied are discussed.     

Effect of isolation on the genetic characteristics of populations in charrs of the genus Salvelinus

To study the effect of isolation on the genetic variation of mDNA, Salvelinus malma krascheninnikovi and S. malma malma nonanadromous populations from different model ecosystems were examined by RT-PCR. The haplotype and nucleotide diversities of mtDNA were significantly decreased in three isolated populations as a result of random gene drift. The genealogy of mtDNA haplotypes was reconstructed. The formation of the modem structure of the isolated populations is discussed in terms of phylogeography.     

Evidence for bimodal hybrid zones between two species of char (Pisces: Salvelinus) in northwestern North America

Dolly Varden (Salvelinus malma, Pisces: Salmonidae) and bull trout (Salvelinus confluentus) have widely overlapping, but largely parapatric ranges in watersheds in northwestern North America from Washington State to northern British Columbia. Genetic analysis of natural populations using diagnostic molecular markers revealed widespread local sympatry and hybridization with hybrids comprising 0-25% of the local samples. In a detailed analysis of hybridization using four nuclear DNA markers and mitochondrial DNA within the Thutade Lake watershed, northcentral British Columbia, hybrid genotypes constituted up to 9% of the population of juvenile char. There were significant deviations from Hardy-Weinberg, gametic, and cytonuclear equilibria, and local samples showed bimodal frequency distributions of genotypes. Pure parental and inferred backcross genotypes were most common, and F1 and F(n) hybrids were comparatively rare. Interspecific hybridization was asymmetrical, with most F1 hybrids (five of six) bearing S. confluentus mtDNA. The introgression of nuclear and mitochondrial alleles was asymmetrical, with S. confluentus mtDNA and Growth Hormone 2 introgressing into S. malma significantly more than either introgression of the three other nuclear loci, or introgression of S. malma alleles into S. confluentus. Substantial prezygotic isolation between the species likely depends on the large body size difference between them in sympatry: S. malma have small bodies and a stream resident life history (12-21 cm adult fork length at maturity), while S. confluentus are larger and adfluvial, i.e., they migrate to Thutade Lake where they grow to maturity before returning to tributary streams to spawn (40-90 cm at maturity). These traits may limit interspecific pairings because of size assortative pairing and size-dependent reproductive habitat use.     

Differentiation of Salvelinus malma charr and Taranetz Charr Salvelinus taranetzi from PCR-RFLP analysis of mitochondrial DNA

Genetic differentiation of two sympatric charr species, the Dolly Varden charr (Salvelinus malma Walbaum) and the Taranetz charr (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 charr and the Taranetz charr are genetically isolated and confirm that S. taranetzi may be regarded as a separate species.     

Introgressive hybridization off chars of the genus Salvelinus as inferred from mitocchondrial DNA variation

Nucleotide sequence analysis of the mitochondrial cyttochrome b gene in the genus Salvelinus revealed hybrids with S. leucomaenis in S. malma malma populations ofthe 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.     

A Rare Case of Permanent Introgressive Hybridization in Char of the Genus Salvelinus (Salmonidae: Salmoniformes) in the Utkholok River,Western Kamchatka

Russian Journal of Marine Biology - Hybrids between the whitespotted char Salvelinus leucomaenis and the northern Dolly Varden S.&nbsp;malma malma have been found to regularly occur and exist...