Molecular Evidence for Multiple Origins of the European Spined Loaches (Teleostei,Cobitidae)

We present a phylogenetic investigation of the Northern Clade, the major monophyletic clade within the freshwater fish family Cobitidae, one of the most prominent families of freshwater fishes found in Asian and European waters. Phylogenetic reconstructions based on the cytochrome b and RAG-1 genes show the genera Microcobitis, Sabanejewia, Koreocobitis and Kichulchoia as monophyletic groups. These reconstructions also show a Cobitis sensu lato and a Misgurnus sensu lato group. The Cobitis sensu lato group includes all species of Cobitis, Iksookimia, Niwaella and Kichulchoia, while the Misgurnus sensu lato group includes Misgurnus, Paramisgurnus and Koreocobitis. Although the monophyly of both the Cobitis sensu lato and Misgurnus sensu lato groups is supported, relationships within the groups are incongruent with current generic definitions. The absence of monophyly of most genera included in the Cobitis sensu lato group (Cobitis, Iksookimia and Niwaella) or their low genetic differentiation (Kichuchoia) supports their consideration as synonyms of Cobitis. Molecular phylogenies indicate that the Asian species of Misgurnus experienced a mitochondrial introgression from a lineage of Cobitis. We also find two nuclear haplotypes in the same Cobitis species from the Adriatic area that, in the absence of morphological differentiation, may indicate molecular introgression. Most lineages within the Northern Clade consist of species found in East Asia. However, some lineages also contain species from Europe and Asia Minor. The phylogenetic relationships presented here are consistent with previous studies suggesting an East Asian origin of the Northern Clade. According to the current distributions and phylogenetic relationships of the Misgurnus sensu lato and Cobitis clade lineages, particularly of M. fossilis and C. melanoleuca, the range expansion of East Asian species into Europe was most likely via Siberia into Northern and Central Europe. Phylogenetic analyses also show that the Cobitis sensu lato group consists of two clear subgroups (I and II), each presenting geographical differences. Subgroup I is distributed exclusively in East Asian drainages with an Eastern European offshoot (C. melanoleuca), whereas Subgroup II includes species widespread throughout Europe (including the Mediterranean), Asia Minor, the Black Sea and the Caucasus, with some lineages related to species restricted to East Asia.     

Molecular phylogeny and intraspecific structure of loaches (genera <Emphasis Type="Italic">Cobitis</Emphasis> and <Emphasis Type="Italic">Misgurnus</Emphasis>) from the Far East region of Russia and some conclusions on their systematics

Russian Far East loaches of the genera Cobitis and Misgurnus are among members of the family Cobitidae with poorly understood systematics. In this study we present phylogenetic hypotheses based on mitochondrial (cytochrome b) and nuclear (RAG-1) sequences. All analyses recovered comparable topological phylogenies, and all data sets supported the non-monophyly of the genera Cobitis and Misgurnus. Both genera are represented by multiple lineages that in some cases do not correspond to the species described. We found some phylogenetic incongruities for the genus Misgurnus (M. mohoity and M. anguillicaudatus) that are explained by ancient hybridization, as was suggested previously for M. anguillicaudatus. The revealed phylogenetic relationships suggest that Paramisgurnus should be treated as a synonym of Misgurnus and M. bipartitus as a synonym of M. mohoity. All analyses recovered C. choii as a member of the genus Cobitis, confirming previous taxonomic conclusions. Most of the molecular lineages found follow currently recognized taxa with some exceptions, such as M. anguillicaudatus and C. lutheri. Phylogenetic relationships recover several unrelated lineages of M. anguillicaudatus and suggest additional studies to solve current taxonomic uncertainty. We found that C. lutheri is a non-natural group that contains two unrelated lineages: specimens of C. lutheri from the Far East of Russia collected close to the type locality and a second lineage with specimens of C. lutheri from Korea, the identification of which must be revised. The study provides evidence of the presence of the Far East species M. nikolskyi in Sakhalin Island, but simultaneously shows conspicuous genetic distinctiveness between the island and the mainland populations.     

Long-term stability of the seasonal succession of different zooplankton species in a brackish water lagoon (southern Baltic Sea)

While it has been widely suggested that freshwater fishes from East Asia invaded the western Palaearctic, details about this process are largely unknown. Here, using the cytochrome b gene, we evaluated the phylogenetic relationships of a small group of Eurasian primary freshwater fishes (Cobitidae), which are widely distributed and species rich in East Asia and Europe, with the purpose of inferring their invasion process of Europe from East Asia. Though phylogenetic relationships of cobitids were not well resolved, our analysis could identify three sister groups formed by the European and East Asian cobitids, which brought new insights into the biogeography of the genera Cobitis, Misgurnus, and Sabanejewia. The present results support the view that Asian cobitid fishes may have invaded Europe at least five times independently, and once reverse colonization of European cobitids to East Asia could also be found. Ancestral Sabanejewia might have been the first cobitids to cross Siberia and invade the EMZS (Euro-Mediterranean zoogeographic subregion) about 33.54 million years ago (MYA). One lineage of Cobitis and the ancestor of Misgurnus fossilis (Linnaeus) almost in the same time invaded the Europe, responding to 16.71 MYA and 16.59 MYA, respectively. Three different lineages of Cobitis were found to have invaded the EMZS from East Asia, and once reverse invasion to East Asia occurred to one subclade of European Cobitis. And our data also suggest that the diversity of East Asian cobitid fishes, especially of the genus Cobitis, is greatly underestimated. Handling editor: C. Sturmbauer     

Evidence of a secondary interspecific mitochondrial DNA introgression in the pond loach Misgurnus dabryanus (Teleostei: Cobitidae) population introduced in Japan

The pond loach Misgurnus dabryanus is a freshwater fish with a distribution range spanning the eastern part of the Asian continent, the Korean Peninsula, and Taiwan. The pond loach was transplanted to the Japanese archipelago through the co-inclusion with dojo loach (Misgurnus anguillicaudatus species complex) populations, which were imported live from China for food materials, and it is currently distributed widely across Japan. A previous mitochondrial DNA (mtDNA) analysis revealed that a pond loach population in Ehime Prefecture (Shikoku Island, Japan) included two highly diverged mtDNA groups (Groups I and II). To examine the origin of these two distinct forms of mtDNA within the Japanese pond loach population, we performed phylogenetic analyses using sequences based on the mtDNA of cytochrome oxidase b (cyt b) and the nuclear DNA recombination activating gene 1 (RAG-1). We also conducted a random amplified polymorphic DNA (RAPD) analysis to examine the establishment of reproductive isolation between sympatric pond loaches with two different mtDNA groups. Our mtDNA phylogenetic results indicated that the two diverged pond loach mtDNA sequences showed polyphyletic relationships among Misgurnus species and its related genus Cobitis. In contrast, there were no clear divergence in nuclear DNA among the pond loaches irrespective of their mtDNA groups, and they all formed monomorphic clades in the phylogenetic relationships among the species. The discrepancy between the mtDNA and nuclear DNA genes support that the existence of two diverged forms of DNA within the pond loach population could be attributed to past mtDNA introgressions from other species rather than convergent evolution. Previous mtDNA phylogenetic studies among Cobitidae revealed that the dojo loach also consisted of two genetically diverged polyphyletic clades: an original Misgurnus mtDNA and an introgressed mtDNA from Cobitis species. In our mtDNA result, the Group II haplotype of the pond loach was included in the mtDNA from the introgressed dojo loach. This suggested that the Group II haplotype was derived from introgressed dojo loach mtDNA. The close relationships between the introgressed dojo loach and the pond loach mtDNA indicated that this secondary introgression had recently occurred via hybridization in a recent artificial aquaculture or transportation process. Common RAG-1 alleles and RAPD bands were shared between the sympatric pond loaches with original and introgressed mtDNAs. This indicates that the introgressed mtDNA haplotype is included as one of the polymorphic genotypes within the pond loach populations, and does not represent existence of different cryptic species.     

Genetic relationships among the Japanese and Korean striated spined loach complex (Cobitidae: Cobitis) and their phylogenetic positions

The striata complex, a group of spined loaches included in the genus Cobitis and characterized by a striped coloration pattern on the lateral midline, is distributed in rivers in northeastern Asia to western Japan. The complex comprises 2 continental species (Cobitis tetralineata and Cobitis lutheri) and 3 Japanese races of species rank (large race, middle race, and small race), the small race further comprising 6 local forms of subspecific rank (Tokai form, Biwa form, Yodo form, Sanyo form, San-in form, and Kyushu form). Previous karyological studies have revealed that the large race is an allotetraploid, the others being diploid. In this study, mitochondrial (mt) DNA analyses were conducted for 30 diploid populations of the Cobitis striata complex from Japan and Korea to examine: (1) their phylogenetic relationships and the position of the complex among the major lineages of Cobitis; and (2) the genetic relationships among the Japanese and Korean populations. The results, based on cytochrome b sequences (724 base pairs) analyzed with those of the main lineage of European and Japanese Cobitis, indicated that the striata complex should be considered as a monophyletic group, which evolved in northeastern Asia. Initially considered as a subspecies of Cobitis taenia, widely distributed from Europe to Asia, the striata complex does not have a sister-relationship with the former. Although the Korean species C. tetralineata was formerly believed to be closely related to the middle race in Japan, and a second continental species, C. lutheri, closely related to the Kyushu or San-in forms of the small race in Japan, the trees resulting from the present study revealed that the two Korean species were clustered with each other and separated from all Japanese races.     

A new loach, Cobitis shikokuensis (Teleostei: Cobitidae), from Shikoku Island, Japan

A new species of spinous loach, Cobitis shikokuensis, is described based on 297 specimens from Shikoku Island, Japan. The new species was formerly known as the Shikoku group of Cobitis takatsuensis. It can be distinguished from other species of Cobitis and closely related genera by a combination of the following characters: dorsal fin with 6 branched soft rays; anal fin with 5 branched soft rays; one brownish streak across eye from the tip of nose, no streak on cheek; a black spot smaller than eye diameter near the dorsal corner of the caudal fin base; 3–5 small brownish speckles on ventral side of caudal peduncle; high caudal peduncle with well-developed fleshy keels on dorsal and ventral side; a lamina circularis at base of dorsal part of pectoral fin absent; first branched soft ray of pectoral fin broad in males; pectoral soft rays widely branched from the approximate midpoint; last anal fin ray with 2 elements; interorbital width 11.2–17.1% of head length.     

Embryonic and larval development of a Japanese spinous loach,Cobitis takatsuensis

The embryonic and larval development ofCobitis takatsuensis, a mountain stream spinous loach, was surveyed by incubating artificially inseminated eggs. The mean diameter of the inflated eggs and mean total length of newly-hatched larvae were 2.7 mm and 5.7 mm, respectively. The eggs were spherical, transparent and unpigmented, with a pale yellow yolk and no oil globule. The daily cumulative temperature to hatching was estimated to be 70–110°C. day. Hatched larvae were unpigmented with outer gill filaments on their cheeks, as in otherCobitis species, but the melanophores were comparatively less obvious at each developmental stage. The larvae started feeding eleven days after hatching yolk absorption being completed sixteen days after hatching. All the fin rays were fully developed and the juvenile stage reached at 16 mm TL, 38 days after hatching. Embryonic and larval developmental traits ofC. takatsuensis, such as egg size, clutch size and larval pigmentation, were similar to the Korean species,Niwaella multifasciata, that lives in the upper reaches of the Nak-tong river, andN. delicata, which inhabits Japanese mountain streams, rather than to its congeners. Among cobitine fishes, the spawning of a small number of larger eggs yielding larger larvae without pigmentation, characteristics shared byC. takatsuensis, N. multifasciata andN. delicata, is attributable to adaptation to cold mountain streams.     

Allozyme divergence between two groups of the Japanese spinous loach, <Emphasis Type="Italic">Cobitis takatsuensis</Emphasis>

Phylogenetic relationships of the Chugoku-Kyushu and Shikoku groups of the Japanese spinous loach, Cobitis takatsuensis, among the diploid congeners in Japan, C. biwae and Cobitis sp. complex with Niwaella delicata as an outgroup, were investigated by analyzing 20 protein-coding loci. The two groups of C. takatsuensis are clearly diverged genetically, with the genetic distance corresponding to the species level (average D=0.27). The two groups form a monophyletic cluster (bootstrap probability, P=94.9%) that is a sister cluster of C. biwae (P=63.1%). The monophyletic cluster of the Cobitis sp. complex (P=93.2%) is the most distantly related in the genus. The present results differ mostly from the mitochondrial phylogeny previously known in which the Chugoku-Kyushu group of C. takatsuensis is connected to the eastern group of C. biwae and the Shikoku group is connected to the Cobitis sp. complex with the western group of C. biwae. The contradiction between the allozymic and mitochondrial phylogenies suggests that the Shikoku group of C. takatsuensis and the western group of C. biwae received mitochondrial introgression from the Cobitis sp. complex.     

Gene mapping of 28S rDNA sites in allotriploid Cobitis females (Pisces: Cobitidae) from a diploid-polyploid population

We performed cytogenetic analysis on triploid Cobitis hybrid specimens from a natural diploid-polyploid population in the Bug River (Poland). Chromosomes were examined by fluorescence in situ hybridization method (FISH) with 28S rDNA as a probe. The rDNA sites were commonly identified in the terminal position of p arms of two small submetacentric chromosomes, in the centromere area of two acrocentric chromosomes and in the terminal position of q arm of one acrocentric chromosome. Some of the observed signals were stronger than others. In few cases, one small acrocentric chromosome showing rDNA sites located terminally on both p and a q arm was observed. This study brings useful information about polymorphism of 28S rDNA sites regarding their number, location and size in triploid specimens. Moreover, it gives preliminary data for comparison with similar researches on the parental species of Cobitis hybrid females, viz. C. taenia, C. taurica and C. tanaitica, and other related Cobitis taxa of different ploidy level. The present study increases our knowledge about ribosomal genes inheritance between Cobitis species and their polyploid hybrids.     

Genetic and environmental variations in quantitative characters in fishes: a comparative analysis of monoclonal triploid and bisexual tetraploid spined loaches (<Emphasis Type="Italic">Cobitis</Emphasis>, Cobitidae)

The heritability estimates of 25 external morphometric characters and 23 craniometric indices are obtained by use of variances in monoclonal all-female triploids and bisexual tetraploids of spined loaches (genus Cobitis, Cobitidae) collected from the same breeding biotope. Most of studied traits demonstrate low heritability confirming previous conclusion on the similarity between external morphometric characters and craniological indices in relative effects of genetic and environmental components in their total phenotypic variation. Low heritability estimates in most of external morphological traits correspond to their low diagnostic value in Cobitis species. As a whole, in spite of certain deviations, studies on clonal forms do not refute the concept on higher heritability estimates in diagnostically significant traits in comparison with traits without diagnostic values in the same taxonomic group. Low heritability in most morphometric traits more probably is resulted from their low additive genetic variation caused by strong selection of evolutionary developed specific body shape in spined loaches, because strong selection should reduce the genetic variance in body proportions to minimal size. Sex differences observed in heritability estimates should be interpreted as a result of linkage of several additive genes controlling these traits to sex chromosomes. A few characters demonstrating high heritability estimates up to 0.492–0.580 are of great interest for taxonomic and phylogenetic studies in genus Cobitis and related taxa.     

Taxonomic Study of Korean <Emphasis Type="Italic">Scirpus</Emphasis> L. s.l. (Cyperaceae) II: Pattern of Phenotypic Evolution Inferred from Molecular Phylogeny

Scirpus L. s.l. is well known as one of the polyphyletic groups in Cyperaceae. Recent molecular phylogenetic studies clearly suggested that Scirpus s.l. should be separated into several independent genera. In this study, we intend to present the morphological variations and patterns of phenotypic evolution based on molecular phylogeny of Korean Scirpus s.l. Five genera, including 21 taxa from Korean Scirpus s.l., were examined: three species of Bolboschoenus, three species of Schoenoplectus, eight species of Schoenoplectiella including one hybrid, five species of Scirpus, and two species of Trichophorum. For morphological analyses, 23 and 48 characters were selected from vegetative and reproductive organs, respectively. Molecular phylogeny was inferred from a nuclear ribosomal internal transcribed spacer, the chloroplast rbcL gene, and the chloroplast trnL-F region. Distinct characteristics and quantitative variation was presented for identification of the five genera and their species. A pronounced pattern of morphological character change was reduction, although many other character states seem to be homoplastic. We suggest that the reduction of phenotypic characteristics has been expressed in terms of condensation of internodes, reduction of leaf blade, and simplification of inflorescence among five genera of Korean Scirpus s.l.     

Chromosomal inheritance of parental rDNAs distribution pattern detected by FISH in diploid F1 hybrid progeny of Cobitis (Teleostei,Cobitidae) species has non-Mendelian character

This study was conducted to describe the major and the minor rDNA chromosome distribution in the spined loach Cobitis taenia (2n = 48) and the Danubian loach Cobitis elongatoides (2n = 50), and their laboratory-produced diploid reciprocal F₁ hybrid progeny. It was tested by fluorescence in situ hybridisation (FISH) whether the number of 28s and 5s rDNA sites in the karyotypes of diploid hybrids corresponds to the expectations resulting from Mendelian ratio and if nucleolar organiser regions (NOR)were inherited from both parents or nucleolar dominance can be observed in the induced F₁ hybrid progeny. Ten (females) or twelve (males) 28s rDNA loci were located in nine uniarm chromosomes of C. taenia. Two of such loci terminally bounded on one acrocentric chromosome were unique and indicated as specific for this species. Large 5s rDNA clusters were located on two acrocentric chromosomes. In C. elongatoides of both sexes, six NOR sites in terminal regions on six meta-submetacentric chromosomes and two 5s rDNA sites on large submetacentrics were detected. The F₁ hybrid progeny (2n = 49) was characterised by the intermediate karyotype with the sites of ribosome synthesis on chromosomes inherited from both parents without showing nucleolar dominance. 5s rDNA sites were detected on large submetacentric and two acrocentric chromosomes. The observed number of both 28s and 5s rDNAs signals in F₁ diploid Cobitis hybrids was disproportionally inherited from the two parental species, showing inconsistency with the Mendelian ratios. The presented rDNA patterns indicate some marker chromosomes that allow the species of the parental male and female to be recognised in hybrid progeny. The 5s rDNA was found to be a particularly effective diagnostic marker of C. elongatoides to partially discern genomic composition of diploid Cobitis hybrids and presumably allopolyploids resulting from their backcrossing with one of the parental species. Thus, the current study provides insight into the extent of rDNA heredity in Cobitis chromosomes and their cytotaxonomic character.     

Comparative histology of the testes of the spined loach Cobitis taenia L. and natural allotetraploids of Cobitis (Pisces, Cobitidae)

The spined loach Cobitis taenia L. creates exclusively diploid and mixed diploid–polyploid populations. Allotriploid females, which co-exist with C. taenia or C. elongatoides and a few tetraploid males and females dominate in most Cobitis mixed populations. They reproduce gynogenetically and produce triploid eggs that are stimulated to development by sperm from Cobitis males. Some of these eggs are fertilized, which leads to the production of bisexual tetraploids. Males of C. taenia (2n = 48) from a diploid population in Lake Klawój, Northern Poland (46 individuals) and from a mixed Cobitis population in the Bug River, Eastern Poland (7 individuals), and three tetraploid males (4n = 98) from the same mixed population were examined. All the fish were analyzed karyologically and histologically. Tubules with cysts of the testes of C. taenia from both populations were filled with germ cells at various developmental stages. Among fishes from Lake Klawój sperm maturation in batches simultaneous with the batch spawning of C. taenia females was found. The testes of the loach C. taenia, from a mixed population in the Bug River, were filled with spermatozoa over the entire reproductive season. Sperm maturation in batches was not observed. Sperm maturation in batches seems to be only connected with a few diploid males in this population. So, a continuous process of spermatogenesis in their testes is required. Only in the testes of all tetraploid Cobitis males were cells characteristic of the early stages of spermatogenesis observed, i.e. without spermatids and spermatozoa. Furthermore, the histological sections of the testis of a male captured in August, revealed fragments with connective tissue between the germ cells. However the participation of tetraploid, infertile Cobitis males in the process of reproduction in the investigated mixed population remains controversial. The results obtained so far, reveal that even the infertile sperm of tetraploid males may induce gynogenesis in Cobitis triploid females.     

Cheek pouch capacities and loading rates of heteromyid rodents

Rodents of the family Heteromyidae are proficient gatherers and hoarders of seeds. A major component of their adaptive specialization for harvesting and transporting seeds is their spacious, fur-lined cheek pouches. Precise measurements of cheek pouch capacities are essential if ecologists are to understand the foraging ecology, possible constraints on locomotion patterns, and competitive relationships of heteromyid rodents. To measure the size of these cheek pouches and the rate at which animals load seeds into their pouches during seed harvest, we attracted 56 individuals representing ten species of heteromyid rodents to bait stations in the field and allowed them to fill their cheek pouches with seeds several times while we observed and timed the events with the aid of night-vision equipment. The largest load taken by each individual was used as an estimate of its cheek pouch capacity. At the end of observations, each subject was captured and its mass and other data gathered. The allometric relationship between cheek pouch capacity and body mass for ten species of heteromyids was significant [pouch capacity (ml) = 0.148 body mass (g)^0.992, r ²=0.91, P<0.0001]. The regression coefficient is ≈1.0, which indicates that the volume of the cheek pouches scales in direct proportion to body size. When the data were subdivided into quadrupeds (Perognathus and Chaetodipus) and bipeds (Dipodomys) (n=5 for each), the relationships between pouch capacity and body mass were significant, but the two regressions were not significantly different from each other. When all loads (full and partial) were considered, subjects filled their cheek pouches an average of 93 ± 10% of pouch capacity (n=185). Cheek pouch capacities from published studies of artificially filled pouches of heteromyids in the laboratory averaged about 40% below the field measurements obtained here. The allometric relationship between mean loading rate and body mass was also significant [seeds/s=1.067 bodymass (g)^0.830, r ²=0.85,P=0.0011), but when quadrupeds and bipeds were considered separately, the relationships were not significant. Seed densities and bulk densities were used to calculate packing coefficients for seed species, which, when used in conjunction with the allometric relationship between cheek pouch capacity and body size, can be used to estimate the maximum load carried by a heteromyid. Except for the very largest kangaroo rat species, a full pouch load of Indian ricegrass seeds represents less than the daily energy requirements of an active heteromyid. Received: 3 March 1997 / Accepted: 15 July 1997     

Taxonomic revision of the genus Jassidophaga Aczél (Diptera: Pipunculidae) in Korea

In this paper, three Korean species of the big‐head fly, genus Jassidophaga Aczél were treated: J. chiiensis (Ouchi), J. pala (Morakote) and J. villosa (Roser). Of these, J. pala is reported in Korea for the first time. A key to Korean species and photographs on external features are given.     

Molecular phylogeny evidence of altitudinal distribution and habitat adaptation in Korean Ephemera species (Ephemeroptera: Ephemeridae)

Molecular phylogeny of the four Korean Ephemera species, Ephemera orientalis, E. sachalinensis, E. strigata, and E. separigata, was inferred from 630 bp sequences of the partial mitochondrial cytochrome c oxidase I (COI) gene. Results indicated that mean intraspecific sequence divergences were 0.70%, whereas mean interspecific divergences were 15.75%, and 17 samples were distinguished to four species correctly by COI sequences. The results also demonstrated that four species of Korean Ephemera assembled a monophyletic group with high support in maximum parsimony and maximum likelihood analyses. This Ephemera group was divided into two major clades of E. orientalis–E. sachalinensis and E. strigata–E. separigata. Furthermore, we demonstrated that this phylogeny explained altitudinal and habitat adaptations of Korean Ephemera species. The E. orientalis–E. sachalinensis clade, a widespread and lowland‐adapted mayfly group, retained plesiomorphic traits such as paired stripes on abdominal segments and was regarded as plesiotypic in terms of habitat adaptation, compared to the geographically more limited and upland‐adapted E. strigata–E. separigata clade.     

DNA barcoding of the South Korean Tettigoniidae (Orthoptera) using collection specimens reveals three potential species complexes

We carried out DNA barcoding on 24 Korean tettigonid species of 19 genera deposited in the National Institute of Biological Resources to reevaluate the preliminary identification of each specimen. Sequence divergence of DNA barcodes obtained from 113 samples of the 24 species ranged from 0 to 30.4%, the intraspecific variation was 0–7.3%, and the interspecific divergence was 1.1–30.4%; we could not examine the barcoding gap. In the neighbor‐joining tree, the branch length among individuals of Tettigonia ussuriana, Paratlanticus ussuriensis, and Hexacentrus japonicus were relatively longer than those in other species. The detailed analysis of the morphological characters and DNA barcodes of the above three species revealed that these three species represent species complexes. The T. ussuriana complex comprised T. jungi, T. uvarovi, and T. ussuriana. Paratlanticus ussuriensis cluster contained four species; one cluster was identified as P. palgongensis based on morphological characteristics, but the other three clusters, including the P. ussuriensis cluster, require further detailed taxonomic analysis. Lastly, two species clusters were identified within the Hexacentrus japonicus clade. Based on the 99% sequence similarity obtained by blast search of the NCBI GenBank database, one of the clusters was identified as H. unicolor. Thus, the DNA barcoding revealed the presence of at least three cryptic species in Korean Tettigoniidae, although more detailed taxonomic analyses are required to establish their status. Therefore, we suggest that DNA barcoding is a very useful tool for increasing the identification accuracy of insect collections.     

Molecular evidence for the taxonomic identity of Korean Adonis (Ranunculaceae)

 Although Korean Adonis has been traditionally recognized as Adonis amurensis Regel and Radde with various infraspecific taxa described, its taxonomic identity is still in dispute. We investigated the genetic variation in 60 individuals from 12 populations in Korea to elucidate the taxonomic identity of the Korean Adonis complex. Random amplified polymorphic DNA (RAPD) analysis revealed that Korean Adonis comprises three species: A. amurensis, A. pseudoamurensis, and A. multiflora. Adonis amurensis is distributed in northern central inland regions of Korea and A. pseudoamurensis is found in southern parts of the Korean peninsula. Adonis multiflora grows only on Cheju Island, which is the southernmost part of Korea. Phylogenetic analysis of nuclear ribosomal internal transcribed spacer (ITS) sequences partially supported the presence of three Adonis taxa in Korea as detected by RAPD analysis. The Adonis population on Jangbong Island located in the West Sea, which was referred to as A. pseudoamurensis on the basis of morphological examination, was separated from the other populations of A. pseudoamurensis. Otherwise, the molecular evidence is well congruent with the recent morphological study that proposes that Korean Adonis consists of these three species. Received: November 6, 2001 / Accepted: February 26, 2002     

Wing and hindlimb myology of vultures and raptors (Accipitriformes) in relation to locomotion and foraging

Herein, we compare functional muscle properties among convergent forms of large, obligate avian scavengers. We performed quantitative analyses of all the muscle masses and cross‐sectional areas (CSA) of the wings and hindlimbs of five species from two families, Cathartidae (New World vultures; Cathartes aura and Vultur gryphus) and Accipitridae (Old World vultures and raptors; Gyps africanus, Aquila rapax, and Buteo lineatus). These species comprise two paraphyletic functional groups, raptors (Aquila and Buteo) and vultures (Gyps, Cathartes, and Vultur). Our ordination analyses based on all of the muscles of the wings, hindlimbs, and wing and hindlimb muscles combined produced patterns that appeared to be more related to phylogeny, with a hint of a functional signal in wing muscle CSA. However, when wing muscles were grouped according to their functional roles (upstroke, downstroke, and wing stabilization), the percentages of mass and CSA allocated to the upstroke and stabilizing muscles were 1.4–5% greater in the vultures than in the raptors. Conversely, when hindlimb muscles were grouped according to their roles in grasping and terrestrial locomotion, the percentages of mass and CSA allocated to grasping muscles were 5.9–14% greater in the raptors. Our results provide a baseline for future lines of inquiry aimed at understanding how muscle mass and CSA are affected differentially across locomotor modules, possibly in response to differential demands on wing and hindlimb function experienced by these disparate accipitriform clades.     

Phylogenetic analysis of six species of pacific abalone (Haliotidae) based on DNA sequences of 16s rRNA and cytochrome c oxidase subunit I mitochondrial genes

Six species of abalones (Haliotidae) are found on the Korean coasts. Identification and characterization of these abalones are usually based on morphologic characters. In this research we compared the partial sequences of the mitochondrial 16S ribosomal RNA and cytochrome c oxidase subunit I genes to identify species using molecular data and to determine their phylogenetic relationships. Sequence alignments and phylogenetic analysis revealed that the 6 species fell into 2 distinct groups which were genetically distant from each other and exhibited little internal phylogenetic resolution. One group included Haliotis discus hannai, H. discus discus, H. madaka, and H. gigantea, while the other group contained H. diversicolor supertexta and H. diversicolor diversicolor. The 16S rRNA sequences were relatively more conserved than to the COI sequences, but both gene sequences provided sufficient phylogenetic information to distinguish among the 6 species of Pacific abalone, and thus could be valuable molecular characters for species identification.