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.     

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.     

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.     

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 RFLI-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 modern structure of the isolated populations is discussed in terms of phylogeography.     

Genetic divergence of populations of the white char <Emphasis Type="Italic">Salvelinus Albus</Emphasis>, northern and southern forms of malma <Emphasis Type="Italic">S. malma</Emphasis> (Salmonidae), by microsatellite loci of DNA

A very small but statistically significant genetic differentiation by allele frequencies (in the Kamchatka basin F _ST = 0.016 and in Kronotskoe Lake F _ST = 0.063) is revealed between sympatric populations of the northern malma Salvelinus malma malma and of the white char S. albus by analysis of nine microsatellite loci. Factor analysis and clusterization of individual multilocal genotypes in the sum sample of sympatric S. malma malma and S. albus also demonstrate their extremely weak genetic differentiation. Divergence between geographically isolated (allopatric) populations of each of these chars exceeds by two-three times the interspecies differentiation in the zones of sympatry. Samples from allopatric populations clusterize by the geographic principle but not by the taxonomic principle. Such a situation may be explained by the absence of reproductive isolation and presence of hybridization between sympatric S. malma malma and S. albus with the simultaneous existence of some limitations of genetic exchange related to the ecological specialization of these chars. Analysis of microsatellite loci revealed highly significant genetic differences between the northern and southern forms of malma S. malma krascheninnikovi by many times exceeding genetic differences between S. malma malma and S. albus. The obtained data do not confirm the independent species status of S. albus but support the opinion of the species level of differences between the northern and southern forms of malma.     

The divergence of the dolly varden char Salvelinus malma in Asian Northern Pacific populations inferred from the PCR-RFLP analysis of the mitochondrial DNA

Genetic differentiation of the dolly varden char Salvelinus malma Walbaum 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.     

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.     

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.     

Occurrence of a hybrid between endemic Miyabe charr Salvelinus malma miyabei and introduced masu salmon Oncorhynchus masou masou in the Lake Shikaribetsu system, Hokkaido, Japan

A hybrid specimen between endemic Miyabe charr Salvelinus malma miyabei and introduced masu salmon Oncorhynchus masou masou was collected in an inlet stream (Yamada Creek) of the Lake Shikaribetsu. This specimen showed intermediate external characteristics between two species and was also confirmed as a hybrid by DNA markers. The mtDNA of the specimen was identical with that of S. malma miyabei, suggesting that the mating occurred between a female Miyabe charr and a male masu salmon. Hybridization with introduced masu salmon may cause deleterious effects on S. malma miyabei.     

On brook chars of the southern Kuril Islands and a possible mechanism of formation of dwarf forms of Dolly Varden trout Salvelinus malma curilus

The specific morphological and biological features of dwarf chars Salvelinus malma curilus from three isolated populations of the southern Kuril Islands Kunashir and Iturup were studied. It was shown that brook dwarfs, in comparison with the migratory form of Salvelinus malma curilus, are characterized by a smaller diameter of spawned eggs (which corresponds to smaller yolk reserves and shorter embryos at hatching), retarded growth and morphogenesis of early juveniles, a mosaic pattern of paedomorphic and peramorphic morphological transformations, and a short lifespan. The population and age variability of the relative sizes and shape of otoliths was revealed. Dwarf populations are typically characterized by considerable morphological differences between males and females corresponding to those in migratory fish. The strategy of dwarfism is assumed to be built into the genome of the migratory ecotype of S. malma curilus.     

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.     

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.     

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.     

On morphological diversity of southern Dolly Varden trout Salvelinus malma krascheninnikovi from water bodies of Sakhalin

Morphological and some biological specific features of nine samples of the southern form of Dolly Varden trout Salvelinus malma krascheninnikovi from water bodies of Sakhalin were studied. Comparative analysis was performed of our own estimates and estimates made by other authors of meristic characters of Sakhalin populations of Dolly Varden trout, as well as of samples from water bodies of the Kuril Islands and Western Kamchatka. Sakhalin populations of S. malma krascheninnikovi are characterized by a low level of morphological differentiation and the absence of geographic structure. No clinal latitudinal variation in sample averages was revealed in either of the nine studied meristic characters, including the number of vertebrae for populations of S. malma krascheninnikovi in the range from Bol’shoi Shantar Island to the south of Sakhalin. The cline in vertebral number from Kamchatka, along the Kuril Islands to Hokkaido, is most likely explained by specific features of distribution and introgressive hybridization at the secondary contact of populations of the northern and southern forms of Dolly Varden trout that originated from different glacial refugia and had a different vertebral number rather than by the temperature gradient.     

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.     

Sympatric Diversification of Dolly Varden Salvelinus malma (Salmonidae) in an Extremely Small Ecosystem

Journal of Ichthyology - A planktivorous form of Salvelinus malma characterized by lacustrine winter spawning was discovered for the first time in north-east Asia in an isolated water system with...     

Charrs of the genus Salvelinus (Salmonidae): hybridization,phylogeny and evolution

Hydrobiologia - Evolutionary history, systematics and taxonomy of charrs of the genus Salvelinus and especially of the representatives of the S. alpinus–S. malma species complex remain...