Ultrastructural study of spermiogenesis in far-eastern redfins of the genus Tribolodon (Pisces: Cyprinidae)

The spermiogenesis and sperm morphology of the Far-eastern redfins Tribolodon hakonensis (Gunther, 1880), T. brandtii (Dybowskii, 1872), and T. sachalinensis (Okada, Ikeda, 1937) were studied using scanning and transmission electron microscopy. At the spermatid stage, ultrastructural analysis of male germ cells showed no obvious differences between these species. The spermatozoa are primitive acrosome-lacking cells, which are typically found in many representatives of Teleostei with external insemination. Unlike other cyprinids, the angle of inclination of centrioles relative to each other differs slightly in the species under study. The mitochondrial apparatus contains one to five (usually one to three) mitochondria in T. hakonensis, two in T. sachalinensis, and one mitochondrion in T. brandtii.     

Genetic divergence of Far Eastern dace species belonging to the genus Tribolodon (Pisces,Cyprinidae) and closely related taxa

Based on a biochemical-genetic approach, heterozygosity and divergence of structural genes of 30 enzyme loci were analyzed in six dace species. In addition, intra- and interspecific divergence of gene expression was analyzed based on a sample of 12 to 15 loci. Mean heterozygosities per individual varied as follows: Tribolodon species, Hobs = 0.007 +/- 0.007 and Hexp = 0.007 +/- 0.007; T. ezoe, Hobs = 0.045 +/- 0.016 and Hexp = 0.067 +/- 0.029. Several variants of genetic distances were estimated. Standard Nei's distances (DN) varied from 0.145 to 0.284 in four dace species studied. As related to Tribolodon dace species, the following genetic distances were obtained for two members of other genera: Pseudaspius leptocephalus, DN = 0.269; Leuciscus waleckii, DN = 0.769. Based on the distance matrices, different clustering algorithms were realized. The main feature shared by different dendrograms was a separate position of the cluster joining Far-Eastern dace species, to which P. leptocephalus and L. waleckii are successively added. Among the species studied, the proportion of loci similar by expression (E) varied from 87 to 100%. The greatest difference was found between anadromous and nonanadromous ecotypes of T. hakonensis, E = 67%. The following conclusions can be made: (1) Four studied species of the genus Tribolodon are rather well genetically differentiated. Diagnostic loci are available. (2) A nominal dace species, T. species, should be considered the fourth isolated species of this genus, which is confirmed by its recent zoological acceptance of this species. (3) The origin and divergence of dace species belonging to the genus Tribolodon are relatively late (1 to 3 Myr ago) historical events. (4) Taxonomically, the genus Tribolodon belong to the tribe Pseudaspinini together with P. leptocephalus, which is confirmed by genetic data. (5) Data on heterozygosity and the divergence of structural and regulatory elements of genome, along with the proposed scheme of speciation types, suggest the following speciation modes for the species studied: for four species, adaptive divergence and for two species, genetic transformation.     

Genetic divergence and phylogenetic independence of Far Eastern species in subfamily Leuciscinae (Pisces: Cyprinidae) inferred from mitochondrial DNA analyses

Classification of freshwater fish in the subfamily Leuciscinae, Cyprinidae is hampered by complexity or lack of morphological diversity. In this study, analyses based on mtDNA sequences were undertaken to clarify phylogenetic relationships among Far Eastern, North American and European species in the Leuciscinae. Evolutionary rate in cytochrome b gene (Cyt-b) and D-loop sequences appear to be almost constant in Leuciscinae. The topology of phylogenetic trees generated by neighbor-joining (NJ) and maximum likelihood (ML) methods based on Cyt-b gene and D-loop sequences was similar. Five major clades, designated clades 1-5, and a minor clade were discriminated. Most of the Far Eastern, North American and European species were included in the major clades. Clade 1, comprised almost entirely of Far Eastern phoxinins, is monophyletic and greatly diverged from the other species of Leuciscinae. From the present phylogenetic relationships and the previous studies, we present the following hypotheses with respect to the evolutionary history of the Far Eastern phoxinins. The Far Eastern species should be classified into Far Eastern-specific genera, although ichthyologists have still insisted that the species should be included in the European genera. The Far Eastern clade 1 consists of two subclades, including genera Pseudaspius-Tribolodon and Far Eastern Phoxinus species. According to our phylogenetic analyses, Pseudaspius leptocephalus and Tribolodon species should be reclassified into the same genus. On the basis of evolutionary rate in Cyt-b gene in Cyprinidae, it is estimated that the Far Eastern lineage diverged approximately 10-14 million years ago (mya) from the common ancestor of Leuciscinae. It is deduced that speciation of the Far Eastern species occurred until approximately 4 mya, in relation to the formation of the Sea of Japan and the Japanese Islands.     

Speciation and dispersal of Tribolodon species (Pisces,Cyprinidae) around the Sea of Japan

Gene products of 22 protein coding loci from 29 populations of a uniquely sea-going cyprinid genus Tribolodon, T. hakonensis (anadromous and/or river-resident), T. brandtii (anadromous), T. ezoe (freshwater) and T. nakamurai (freshwater), were analyzed in order to infer the genetic divergence and dispersal of Tribolodon species around the Sea of Japan. According to the genetic distance, T. brandtii is located near the center between T. hakonensis and T. ezoe-T. nakamurai cluster. The outgroup root (Pseudaspius leptocephalus, Rhynchocypris lagowskii and Leuciscus waleckii) is attached between T. hakonensis-T. brandtii and the T. ezoe-T. nakamurai clusters. The origin of the genus Tribolodon and its salinity tolerance may date back to the Miocene Sea-of-Japan Lake and the Pliocene Paleo-Sea-of-Japan, respectively, as judged from the divergence time between Tribolodon and the outgroups. The genetic relationships and distribution pattern of Tribolodon species suggest they speciated on the Sakhalin-Japan side, and the continental populations of T. hakonensis and T. brandtii originated from anadromous colonizers. A new faunal element of Japanese freshwater fishes, the Sakhalin-Japan sub-element, is proposed for Tribolodon and some other fishes.     

Cephalic lateral line systems in the Far Eastern species of the genus Phoxinus (Cyprinidae)

The cephalic lateral line systems of seven Far Eastern Phoxinus species (P. phoxinus, P. kumgangensis, P. semotilus, P. lagowskii, P. oxycephalus, P. perenurus, and P. czekanowskii) were investigated. In this genus, the infraorbital canal is not connected with either the supraorbital canal or the preoperculomandibular canal. Phoxinus phoxinus is unique for having an underdeveloped condition, such as canal formation or remaining as uncovered. A unified supraorbital canal was observed in all species, but the infraorbital canal of both P. perenurus and P. czekanowskii was not unified into a single canal throughout their development. Unification between both sides of the supratemporal canal occurred in larger individuals of P. lagowskii, P. oxycephalus, and P. czekanowskii. The preoperculomandibular canal of P. kumgangensis, large P. lagowskii, and large P. oxycephalus was unified. The pore number of each part of the canal system also varied depending on the species. Intraspecific variations were observed between Korean and Japanese specimens of P. lagowskii in the unification of the supratemporal canal and the preoperculomandibular canal, and the number of pores of the supratemporal canal. It was inferred that the specific characteristic patterns of their cephalic lateral line systems reflected the following two factors: different environmental requirement for their microhabitat and different maximum body size.     

The Formation of Biological Diversity of Far Eastern Redfins of the Genus Tribolodon (Cyprinidae)

Russian Journal of Marine Biology - An evolutionary scenario of the origin and distribution of Far Eastern redfins of the genus Tribolodon, based on the data of their phenotypic and biological...     

Phylogeographic analysis reveals two periods of divergence in large-scaled redfin Tribolodon hakonensis (Pisces, Cyprinidae)

Analysis of mtDNA variation in one of the amphidromic Far Eastern redfins, Tribolodon hakonensis, revealed the presence of three considerably genetically different mtDNA phylogroups in the individuals from the Russian part of the range. These data suggest the presence of the two periods of divergent evolution in the history of the species examined. Comparison of the haplotype distributions from different phylogroups over the species range revealed geographic localization of only one phylogroup in the population samples from southern continental coastal regions of the Sea of Japan. At the same time, two other phylogroups were found in almost equal ratios in northern continental samples and near the Sakhalin Island. These results suggest that the first stage of the divergent evolution, which occured between Pliocene and Pleistocene, resulted in the formation of genetically isolated form (probably, a species) in the region of the Sea of Japan. The second, later period of divergence, probably associated with the separation of the Sea of Okhotsk from the Pacific Ocean then ended with the integration of earlier genetically separated forms into one species with the common gene pool.     

Differentiation of large-scaled redfin Tribolodon hakonensis (Pisces, Cyprinidae) in the Russian part of the range as inferred from the data of karyological analysis and PCR-RFLP analysis of mitochondrial DNA

Considerable differences in karyotypes of Tribolodon hakonensis from Primorye and the rivers of the Sea of Okhotsk drainage were demonstrated. These differences raise doubts that these fishes belong to one species and point to the necessity of more precise definition of the species status of the southern form of T. hakonensis. The karyological evidence is consistent with the data of mtDNA PCR-RFLP analysis on genetic independence of the southern and the northern forma of T. hakonensis. In the northern form of T hakonensis, variation of the chromosomal arm number was revealed. Specifically, the number of chromosomes was constant (2n = 50), whereas the number of chromosomal arms (NF) constituted 88, 92, and 94, which suggests genetic heterogeneity of the northern form. PCR-RFLP analysis of mtDNA showed that haplotypes of northern T. hakonensis split into two groups with 100% support. Based on comparative phylogenetic and karyological analyses of the Tribolodon species, independent divergence of the southern and the northern forms of T. hakonensis was suggested.     

Morphogenesis of the pharyngeal teeth in the japanese dace,Tribolodon hakonensis(Pisces: Cyprinidae)

The appearance pattern of pharyngeal tooth germs was investigated in the larval Japanese dace, Tribolodon hakonensis, which has a bilaterally asymmetrical dentition. Teeth develop in a series of replacement waves beginning with the initial central tooth (Ce) and continuing with teeth of anterior (An) and posterior (Po) positions relative to the initial one. Identified by wave number (n) and tooth position (r), according to the formula _n-1[r], tooth germs appeared in the order of tooth ₀[Ce0], ₁[Po1], ₁[Anl], ₂[Ce0], ₂[An2], ₃[Po1], ₃[An1], ₄[Ce0], 4[An2], ₅[Po1], ₅[An1], ₅[An3], ₆[Ce0], ₆[An2] during the larval period. Dentition on the right side, however, lacks the first tooth at position An2 (tooth ₂[An2]) and teeth at position An3. Tooth germs on the first, second, and third replacement waves appeared simultaneously on the arches of both sides. During following waves, tooth germs on the left side appeared later than those on the right. Delay of tooth germ appearance On the left side is interpreted as an inhibitory influence of existing tooth germs in accordance with Osborn's (Proc. R. Soc. Lond. Ser. B 179:261--289, '71) theory. The delay of tooth germ appearance on the left arch is most pronounced on the seventh replacement wave. Teeth of the right major row in adults of this species are replaced more frequently than those of the left major row, apparently in correlation with the absence of the first larval tooth at position An2 and teeth at position An3. It is hypothesized that cyprinids evolved the minor rows and specialized teeth of their adult dentition as apomorphic characteristics by the process of neoteny.     

Nucleotide sequence variation of the mitochondrial COI gene in several eelpout species of the genus Zoarces (Zoarcidae, Pisces)

The mitochondrial COI gene was for the first time sequenced in eelpout species of the genus Zoarces from the Taui Bay of the Sea of Okhotsk: notched-fin Z. elongatus Kner, 1868 and the new sympatric form Zoarces sp., differing in a large set of morphological traits. The two species were compared with European eelpout Z. viviparus Linnaeus, 1758 from the Finnish Bay of the Baltic Sea. Divergence and phylogenetic analyses demonstrated a greater genetic similarity between Zoarces sp. and Z. viviparous than between sympatric Zoarces sp. and Z. elongatus.     

Nucleotide sequence variation of the mitochondrial COI gene in several eelpout species of the genus Zoarces (Zoarcidae, Pisces)

The mitochondrial COI gene was for the first time sequenced in eelpout species of the genus Zoarces from the Taui Bay of the Sea of Okhotsk: notched-fin Z. elongatus Kner, 1868 and the new sympatric form Zoarces sp., differing in a large set of morphological traits. The two species were compared with European eelpout Z. viviparus Linnaeus, 1758 from the Finnish Bay of the Baltic Sea. Divergence and phylogenetic analyses demonstrated a greater genetic similarity between Zoarces sp. and Z. viviparus than between sympatric Zoarces sp. and Z. elongatus.     

Local and global multivariate analysis of geographical mitochondrial DNA variation in Leuciscus cephalus L. 1758 (Pisces: Cyprinidae) in the Balkan Peninsula

Geographical variation of cytochrome b mitochondrial DNA (mtDNA) in chub (Leuciscus cephalus L.) was analysed in 31 samples from 20 different river basins in the Balkan Peninsula and Danube catchment. Multivariate methods of ordination were used to analyse variation of the data sets. The results were interpreted in the context of the proposed ichthyogcographic districts separating the Balkan Peninsula into two main ichthyogeographic divisions (Eastern Greece/Ponto-Acgcan and Western Greece/South Adriatic-Ionian). Boundary detection supported these two ichthyogeographic districts for L. cephalus , revealing a boundary that ran from north to south through the Balkan Peninsula and the middle of Greece. The results also revealed the existence of a third division in Central Greece. The results of ordination techniques on homogeneous zones and analysis of the molecular variance confirmed the results obtained in studying local variability (boundaries). They also allowed us to test the existence of possible subdivisions proposed by different authors inside the two main ichthyogeographic districts. These subdivisions were not supported. The multivariate methods used in this study allowed us to propose a coherent picture of chub ichthyogeographic districts in terms of boundary detection and maximal autocorrelation between populations and to explain the patterns of chub mtDNA variation. A complete interpretation of results concerning L. cephalus requires careful consideration of both boundary analysis and autocorrelative approach. Results from an the autocorrclativc approach alone could lead to substantial misinterpretations.     

Phylogeography of Altai osmans (Oreoleuciscus sp., Cyprinidae, Pisces) based on sequence variation of the mitochondrial DNA cytochrome b gene

Representatives of the genus Oreoleuciscus (Altai osmans) from the water bodies of Western Mongolia were examined for sequence polymorphism of the mitochondrial DNA fragment containing the cytochrome b (Cyt-b) gene. A total of 17 sequence variants (haplotypes) were discovered, which formed two clusters, A and B, with nonoverlapping geographic localization. Cluster A included haplotypes of Altai osmans from the Valley of Lakes water bodies. Cluster B consisted of two subclusters, the first of which (B1) united sequence variants of the populations from Hollow of the Great Lakes and the lakes of the Great Altai Range. The second subcluster (B2) was formed by the haplotypes originating from the Hollow of the Lake Uvs, some isolated lakes of the Northwestern Khangay region, and the water bodies of the Arctic Ocean Basin (basins of Selenga and Orchon rivers). Based on the genetic divergence estimates and the radiation time of genetic geographic groups identified, the existence of three allopatric species of Altaic osmans in the genus Oreoleuciscus was substantiated.     

Phylogeography of Altai osmans fishes (Oreoleuciscus sp., Cyprinidae, Pisces) inferred from nucleotide variation of the mitochondrial DNA cytochrome b gene

In this study, analyses based on cytochrome b gene (Cyt-b) mtDNA sequences were undertaken to clarify phylogenetic and phylogeographical relationships among populations Oreoleuciscus sp. (Pisces: Cyprinidae) from Western Mongolia. Two major clusters, designated clusters A and B, and minor clusters were discriminated. Cluster A comprised specimens of Oreoleuciscus from the Valley of Lakes. The cluster B consists of two subclusters; one of them (B1) including populations Oreoleuciscus of the Hollow of the Big Lakes and lakes Big Altai Range. Second subclusters (B2) combines specimens of Oreoleuciscus from Hollow of the Lake Uvs, some isolated lakes of the North-Western Khangay and waterbodies of the Arctic Ocean Basin (Selenge and Orchon basins). On the basis of estimations of sizes genetic and time of a divergence of the revealed genetikal-geographical groups existence of three allopatric species of the within genera Oreoleuciscus is proved.     

Genetic Divergence of Far-Eastern Dace Species Belonging to the Genus Tribolodon (Pisces, Cyprinidae) and Closely Related Taxa

Based on a biochemical–genetic approach, heterozygosity and divergence of structural genes at 30 enzyme loci were analyzed in six dace species. In addition, intra- and interspecific divergence of gene expression was analyzed based on a sample of 12 to 15 loci. Mean heterozygosities per individual varied as follows: Tribolodon species, H _obs = 0.007 ± 0.007 and H _exp = 0.007 ± 0.007; T. ezoe, H _obs = 0.045 ± 0.016 and H _exp = 0.067 ± 0.029. Several variants of genetic distances were estimated. Standard Nei's distances (D _N) varied from 0.145 to 0.284 in four dace species studied. As related to Tribolodon dace species, the following genetic distances were obtained for two members of other genera:Pseudaspius leptocephalus, D _N = 0.269; Leuciscus waleckii, D _N = 0.769. Based on the distance matrices, different clustering algorithms were realized. The main feature shared by different dendrograms was a separate position of the cluster joining Far-Eastern dace species, to whichP. leptocephalus and L. waleckii are successively added. Among the species studied, the proportion of loci similar by expression (E) varied from 87 to 100%. The greatest difference was found between landlocked and landlocked ecotypes of T. hakonensis, E = 67%. The following conclusions can be made: (1) Four studied species of the genus Tribolodon are rather well genetically differentiated. Diagnostic loci are available. (2) A nominal dace species, T. species, should be considered the fourth unambigous species of this genus, which is confirmed by its recent zoological acceptance of this species. (3) The origin and divergence of dace species belonging to the genus Tribolodon are relatively late (1 to 3 Myr ago) historical events. (4) Taxonomically, the genus Tribolodon belong to the tribe Pseudaspinini together with P. leptocephalus, which is confirmed by genetic data. (5) Data on heterozygosity and the divergence of structural and regulatory elements of genome, along with the proposed scheme of speciation types, suggest the following speciation modes for the species studied: for four species, adaptive divergence and for two species, genetic transformation.     

Taxonomic differences and ecological conditionality of scale structure in three morphologically similar species of Far Eastern redfins of the genus <Emphasis Type="Italic">Tribolodon</Emphasis> (Teleostei: Cyprinidae)

Using the example of three morphologically similar species of the Far Eastern redfins of the genus Tribolodon, it has been shown that an analysis of scale structure allows species identification of not only adults, when they are not in their characteristic spawning dress, but also immature individuals. The most distinct interspecies differences of redfins have been found in the case of the combined use of three basic parameters of scales: lesser diameter, greater diameter, and number of radii. The differences between redfins in the shape and structure of their scales can be explained by the features of ecology of these species, e.g. the velocity of water flow in the breeding grounds and during overwintering in rivers.     

Hexaploidy in yellowfish species (Barbus, Pisces, Cyprinidae) from southern Africa

Five small-scaled yellow fish (large Burbus spp.) from southern Africa are shown to have modal 148 or 150 chromosomes. Themajority ofcyprinid species have 2N = 50 chromosomes, indicating that the yellow fish karyotype is hexaploid in origin. However, as there is no indication that the species are unisexual or that normal reproduction occurs by any means other than bisexual fertilization, the yellowfish karyotype is considered to have reverted to a diploid condition.