No reduction of hatching rates among F1 hybrids of naturally hybridizing three Far Eastern daces,genus <Emphasis Type="Italic">Tribolodon</Emphasis> (Cypriniformes,Cyprinidae)

Egg hatching rates of F1 hybrids among three fishes, Japanese dace Tribolodon hakonensis, Pacific redfin T. brandtii, and rosyface dace T. sachalinensis, were compared with pure crosses. They are highly divergent (diverged 10–20 million years ago) but naturally hybridizing. Although crosses involving female rosyface dace were not available, hatching success was high among all combinations of hybrids (76 ± 23%) and comparable to pure crosses (86 ± 8%), implying ecological or genetic isolation mechanisms maintaining each species in later life stages.     

Comparative phylogeography of diadromous and freshwater daces of the genus <Emphasis Type="Italic">Tribolodon</Emphasis> (Cyprinidae)

Far Eastern daces, genus Tribolodon (Cyprinidae), are thought to have diversified and developed unique diadromous life histories under changing conditions in the Sea of Japan and the surrounding environment. To examine the relationships between life history traits, distribution, and genetic population structures, we conducted a comparative phylogeographic analysis using partial mtDNA sequence data from samples collected over almost the full ranges of all four Tribolodon species. Phylogenetic analyses revealed several intraspecific haplotype groups that differentiated in the early Pleistocene to the Pliocene with or without geographic overlaps. A time-calibrated phylogeny suggested that the relatively smaller geographic ranges of the strictly freshwater species, T. sachalinensis and T. nakamurai, were explained not by the recent origins of these species, but by their limited dispersal abilities and smaller historical population sizes. The wider-ranging diadromous species, T. brandtii and T. hakonensis, exhibited similar major phylogeographic structures in their distributions, but the chronological order and timing of formation of this structure largely differed between the two species. In addition to those differences, the overlapping patterns of the differentiated intraspecific lineages in these species suggest dynamic, but somewhat restricted dispersal during the Plio-Pleistocene. Tribolodon hakonensis, one of the most widespread species of East Asian freshwater fishes, included both common and unique phylogeographic patterns compared to other fish species; the unique patterns (i.e., its wide range across freshwater biogeographic boundaries like the sea and mountains) would reflect its ecological features as a remarkable generalist inhabiting lakes, upper and lower reaches of rivers, and even coastal areas.     

A taxonomic study of the genusCalamagrostis on Mount Yakeishidake

Populations of seven species ofCalamagrostis growing on Mt. Yakeishidake were studied from a viewpoint of taxonomy. Four of them (C. sesquifolia, C. fauriei, C. matsumurae, C. gigas) were composed of tetraploid plants with 2n=28 and high pollen fertility.C. matsumurae andC. gigas had sympatric contacts, but no sign of their hybridization could be detected. The population ofC. sachalinensis on this mountain included hexaploids (2n=42) and octoploids (2n=56), and no obvious morphological or ecological differences could be found between them. The population ofC. sachalinensis was estimated to include various ecological races which may be continuously graded. InC. langsdorffi, only octoploid plants were found. Plants with 2n=56+0?1B were found in the population ofC. hakonensis. The pollen features of the collections ofC. langsdorffii, C. sachalinesis andC. hakonensis showed that apomictic seed formation may be more or less developed in them. Populations of these species on Mt. Yakeishidake were clearly separated morphologically. It was concluded that the populations of the seven species examined each have a unity and that strains ofC. langsdorffii, C. sachalinensis andC. hakonensis found on Mt. Yakeishidake are confined to those of a derived type.     

Otolith Sr : Ca ratios of the freshwater and anadromous cyprinid genus Tribolodon

The ratio of strontium to calcium concentration in the otolith of four species of Tribolodon (Cyprinidae) was examined to reveal their migratory histories. The ratio was relatively stable from the core to the edge of the otolith in fluvial Tribolodon ezoe and T. hakonensis, whereas it changed from a low level to a higher level in anadromous T. brandtii and T. hakonensis. Four specimens of T. nakamurai from the Mogami River, Yamagata Prefecture, northern Japan, showed relatively stable ratios, indicating a fluvial life mode. On the other hand, the ratio increased once in only one specimen of T. nakamurai from the Koyoshi River of Akita Prefecture, suggesting the possibility that it was born in the Mogami River, the nearest major habitat of this species, and accidentally migrated to the Koyoshi River through the sea.     

Analysis of mitochondrial DNA: Taxonomic and phylogenetic relationships in two fish taxa (Pisces: Mugilidae and Cyprinidae)

To solve some systematic questions as well as to study genetic variability and evolutionary relationships in two groups of fish belonging to the Mugilid (Mugilidae) and Cyprinid (Cyprinidae) families, we have used restriction fragment length polymorphism analysis of mitochondrial DNA (mtDNA) fragments amplified in polymerase chain reaction. The analysis of three mtDNA fragments of 7220 bp total length of six Mugilid species has shown that Mediterranean Liza aurata, L. ramada, L. saliens, and Chelon labrosus form a common cluster, L. aurata and C. labrosus being the closest relatives, whereas L. haematocheilus (syn. C. haematocheilus) of the Sea of Japan forms a sister group to the Mediterranean cluster. It was found that Chelon and Liza genera are paraphyletic, and therefore their division into two genera is unnatural and they should be synonymized. According to priority, Liza species should be ascribed to Chelon genus. Mugil cephalus is the most distant compared to the rest of the species studied. The level of genetic divergence between allopatric samples of M. cephalus from the Sea of Japan and the Mediterranean Sea has proved to be very high—4.5% of nucleotide substitutions. The analysis of four mtDNA fragments of 9340 bp total length of six Cyprinid species has shown that L. waleckii is the most genetically distant. Pseudaspius leptocephalus is a sister group to Tribolodon species. All Tribolodon species form a common cluster with T. sachalinensis as a root. The remaining species form two branches, one of which includes T. nakamurai and T. brandtii, another one combines T. hakonensis and a new form of Tribolodon revealed that is close to T. hakonensis by its mtDNA (2.4% of nucleotide substitutions). This new form might be an independent species. Published in Russian in Biokhimiya, 2007, Vol. 72, No. 12, pp. 1651–1658.     

Differentiation of large-scaled redfin <Emphasis Type="Italic">Tribolodon hakonensis</Emphasis> (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 forms 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.     

Phylogenetic analysis of Altai osmans of the genus <Emphasis Type="Italic">Oreoleuciscus</Emphasis> (Pisces,Cyprinidae, Leuciscinae), based on the analysis of the cytochrome oxidase 1 gene (<Emphasis Type="Italic">Co-1</Emphasis>) sequence

Molecular genetic analysis of Altai osmans of the genus Oreoleuciscus from two different parts of the range was carried out. In this study, based on the mitochondrial Co-1 gene sequence, a total of 25 fish specimens belonging to four genera were examined: (1) O. humilis, 2 specimens; O. potanini, 13 specimens; (2) Pseudaspius leptocephalus, 1 specimen; (3) Tribolodon brandtii, T. hakonensis, and T. sachalinensis from the GanBank database, 9 speciens; and (4) Leuciscus waleckii, 1 specimen (used as an outgroup). The p-distances were very low both within and between the species: (1) 0.20 ± 0.03%; (2) 0.40 ± 0.12%; and (1–2) 0.80 ± 0.04%. To visualize the relationships among all of the species examined, the neighbor joining (NJ), maximum parsimony (MP), Bayesian (BA), and maximum likelihood (ML) trees were constructed. The results obtained using these methods were very similar. It was demonstrated that species assignment of the individuals (barcoding) with the help Co-1 gene was effective, despite of very low divergence of the two osman taxa, which was comparable with typical intraspecific values in other animal groups. Taxonomic status of O. potanini and O. humilis requires further investigation with paying attention to low genetic distances between these species along with the lack of material from sympatric parts of the ranges.     

Comparative morphological analysis of big-scaled redfin <Emphasis Type="Italic">Tribolodon hakonensis</Emphasis> Günther, 1877 (Cyprinidae,Cypriniformes) from Sakhalin and southern Primorye

Comparative analysis of plastic and meristic characteristics of big-scaled redfin Tribolodon hakonensis from Sakhalin and southern Primorye has been performed. The considerable differences revealed are supported by results of genetic studies. The results of comparison of morphology of big-scaled and Pacific T. brandtii redfins with the purpose of assessing the probability of the previously advanced hypothesis on the origin of the “southern” form of big-scaled redfin are provided.     

Web image search revealed large-scale variations in breeding season and nuptial coloration in a mutually ornamented fish, <Emphasis Type="Italic">Tribolodon hakonensis</Emphasis>

Geographic variations in reproductive traits are important for evolutionary biology, but often difficult to investigate because of the need for a large-scale survey and the ephemeral nature of secondary sexual characteristics. Here, using web image searches (Google Images and Twitter), we revealed large-scale geographic variations (>1500 km) in breeding timing and nuptial coloration in the mutually ornamented fish, Japanese dace (Tribolodon hakonensis). As this fish is easily caught and many anglers upload their photographs on the web, we were able to find a total of 401 high-resolution photographs from all over Japan. Breeding periods were determined from dates of photographs with/without nuptial coloration, which matched with previous studies. We also found that breeding periods might have advanced three weeks over the last 80 years in the Chitose River, potentially because of climate change. Additionally, the pattern of latitudinal cline for breeding timing revealed delayed timing in higher latitudes, although regional variations were also high. Finally, we quantified the patterns of nuptial coloration for this mutually ornamented fish, and confirmed that over 80% of individuals showed typical colorations, but others showed rare phenotypes that were never previously described, including an intermediate coloration between T. hakonensis and a potentially hybridizing species, T. brandtii. Our web-based method for estimating breeding phenology could be adopted for organisms with temporal sexual characteristics, such as plants (e.g., flowering time) and some fishes. Web image analyses are still preliminarily with many limitations, but could be promising for investigating variations in visible traits.     

Single and group organizations of individual animals in the community of bats (Chiroptera: Vespertilionidae) hibernating in artificial caves of the Samarskaya Luka

The spatial arrangement of individual bats of seven species and their tendency to form groups were studied in artificial caves of the Samarskaya Luka during five winter periods (1999 and 2003–2006). It was demonstrated that formation of dense groups did not depend on the hibernating population size or density and was related to the biological characteristics of the species. Most Myotis brandtii and M. dasycneme (60–80% of the populations) wintered in groups. M. mystacinus, M. daubentonii, Plecotus auritus, M. nattereri, and Eptesicus nilssonii usually wintered singly (87% of the animals). In M. brandtii and M. dasycneme, as many as 78% of individual animals were in conspecific groups, whereas this proportion for the other species was no higher than 11%. All pairs of species exhibited negative assortativeness, which indicated a higher preference of spending winter in conspecific groups than in groups containing representatives of other species. Estimation of the ratios between groups containing animals of one and several species showed that conspecific groups were prevailing in M. brandtii and M. dasycneme, whereas all other species usually formed mixed groups.     

Current state of knowledge of the Korean Cynipini: subspecies description,new combinations and checklist (Hymenoptera,Cynipidae)

Prior to this study, only 8 species of oak gall-forming wasps were known from Korea. After material examination and an exhaustive bibliographic revision, a total of 17 oak gall wasps species have been reported. The following species correspond to new records: Andricus hakonensis (Ahmead); A. pseudocurvator Tang & Melika; A. songshui Tang & Melika; Cerroneuroterus folimargo (Monzen); C. monzeni (Dettmer); C. vonkuenburgi (Dettmer); Neuroterus hakonensis Ashmead, Trichagalma acutissimae (Monzen), T. formosana Melika & Tang. It was impossible to obtain adult specimens for all the species, and and the following species are recorded only based on galls: Andricus kashiwaphilus Abe; A. mukaigawae (Mukaigawa); A. pseudoflos (Monzen); A. targionii Kieffer; Cerroneuroterus folimargo (Monzen); C. monzeni (Dettmer); Neuroterus hakonensis Ashmead; Trichagalma acutissimae (Monzen). The species Dryocosmus kunugiphagus Ide & Abe, 2015 is transferred to the genus Plagiotrochus: Plagiotrochus kunugiphagus (Ide & Abe, 2015) n. comb. Additionally a new subspecies of Trichagalma formosana is described to include the material from Korea and Mainland China, closely related but morphologically different to the Taiwan populations. For each species live cycle, morphology of galls, hosts, phenology, and distribution are commented. In some of them taxonomic or morphological comments are mentioned and a key of agamic Trichagalma with dark smoky pigmented spots in the forewing is added. All galls are illustrated. An up-to-date checklist of the Korean Cynipini is provided. The previously reported species from Korea (Andricus noliquercicola Shinji, 1938; Aphelonyx crispulae Mukaigawae, 1920; Neuroterus narae Shinji, 1941 and Neuroterus nawai Ashmead, 1904) are here excluded from the Korean species list.     

Seed surface morphology in some representatives of the Genus Rhodiola sect. Rhodiola (Crassulaceae) in the Russian Far East

The seed coat morphology and anatomy of Rhodiola angusta, Rh. alsia, Rh. integrifolia, Rh. ishidae, R. rosea subsp. arctica, Rh. rosea subsp. rosea and Rh. rosea subsp. sachalinensis were examined comparatively using scanning and light microscopy methods in order to evaluate their characteristics for use in systematic studies. Based on the features of the arrangement of cells and cell outline, three morphological types of seed coats were identified – laticostate (Rh. rosea subsp. rosea), tenuicostate (Rh. integrifolia, Rh. rosea subsp. rosea R. rosea subsp. arctica and Rh. rosea subsp. sachalinensis) and colliculate (Rh. alsia, Rh. angusta, Rh. ishidae). Exotesta sculpture features are stable across a range of habitats in all taxa except Rh. rosea subsp. rosea. Considerable variability of seed coat morphology is reported in the latter species and its possible implication for species taxonomy is discussed.     

The karyotype of the great sculpin, <Emphasis Type="Italic">Myoxocephalus polyacanthocephalus</Emphasis> (Pallas, 1814) (Pisces: Cottidae), from the Russian part of the species range

The karyotype of the great sculpin, Myoxocephalus polyacanthocephalus (Pallas, 1814) (Pisces: Cottidae) from the Sea of Okhotsk and the Sea of Japan has been studied for the first time. The karyotype is stable; it consists of 40 chromosomes (4 metacentric, 2 submeta-subtelocentric, 20 subtelocentric, and 14 acrocentric chromosomes); the number of chromosomal arms is 44 + 2. Nucleolar organizer regions (NOR) are found in the telomeric region of the arm in one homologue of a pair of small metacentric chromosomes, using the Ag-NOR banding technique. A comparative analysis of the karyotype of M. polyacanthocephalus and the karyotypes of other Myoxocephalus species (M. stelleri, M. brandtii, M. jaok, M. ochotensis, and M. scorpius) has been carried out based on the main karyotype characters, as well as on the number and localization of NORs. The identified differences make it possible to differentiate the studied species, whereas the general traits indicate their taxonomic proximity.     

Floral morphology and systematics of the genus Thottea Rottb. (Aristolochiaceae) from the Western Ghats, India

The medicinal genus Thottea (Aristolochiaceae) is represented in the Western Ghats, India, by eight species, seven of which are endemic to this region. In the present study, diversity in floral structure and organization of these eight species was analyzed from a systematic perspective. Floral morphology was thoroughly studied and discussed with emphasis on 16 qualitative and 11 quantitative characters. Statistical tools such as UPGMA cluster analysis, PCoA, PCA, and one-way ANOVA were used for elucidating species boundaries and inter-relationships. Significance of the selected qualitative and quantitative characters for species level systematics was revealed by the PCA and one-way ANOVA. Based on the nature of sepals (free/fused), two species groups were identified (first group with T. duchartrei and T. idukkiana, and the second group with the remaining six species). By considering the number and arrangement of stamens, yet another species grouping was proposed (one with T. abrahamii and T. dinghoui, and the other with the remaining six species). The UPGMA phenogram and the PCoA scatter plot clearly pointed out the existence of two species groups: the first group with T. abrahamii, T. barberi, T. dinghoui, T. ponmudiana, T. siliquosa, and T. sivarajanii, and the second one with T. duchartrei and T. idukkiana. With regard to the morphometric tools and the taxonomic key, it does not seem possible to distinguish T. abrahamii from T. dinghoui, and T. idukkiana from T. duchartrei.     

Differential photosynthetic characteristics between seedlings and saplings of Abies sachalinensis and Picea glehnii, in the field

Abies sachalinensis and Picea glehnii are co-dominant tree species and major components of the forests of Hokkaido, Japan. Recent work suggests that a reversal in potential competitive superiority at different developmental stages could be important to explain their coexistence. Such shifts in competitive advantage can be mechanistically understood by studying the corresponding physiological differences between distinct life stages. Accordingly, our objective was to investigate in the field the photosynthesis of shade-growing juveniles of these species from two different size-classes, seedlings and saplings. Our results show that seedlings of both species had higher concentrations of photoprotective xanthophylls than saplings, especially in spring, and suggest that seedlings have a lower threshold of stress tolerance than saplings. Photosynthetic capacity per needle area and lateral shoot extension rate decreased from the seedling to the sapling stage in A. sachalinensis, while in P. glehnii, both increased from the seedling to the sapling stage. Abies sachalinensis had higher photosynthetic rates at the seedling stage but lower rates at the sapling stage than P. glehnii. Nevertheless, A. sachalinensis had a higher lateral shoot extension rate than P. glehnii at both stages. Our physiological results support previous ecological observations that A. sachalinensis is a superior competitor to P. glehnii in the understory, and show that its competitive advantage is higher at the seedling stage than at the sapling stage.     

Post‐glacial history and introgression in Abies (Pinaceae) species of the Russian Far East inferred from both nuclear and cytoplasmic markers

Aim The main aim of the present study is to infer the post‐glacial history of Abies species from north‐east Asia and to test the hypotheses that coastal Abies populations suffered less from climatic fluctuations during Pleistocene glacial periods than their more continental counterparts, and that Sakhalin was a major area of introgression. Location Natural ranges of the fir species Abies nephrolepis, Abies sachalinensis and Abies holophylla in the Russian Far East, and of Abies gracilis, which is endemic to the Kamchatka Peninsula. Methods Nineteen populations were sampled for allozyme analysis. Seventeen of these populations were also screened for variation at two paternally inherited chloroplast DNA microsatellite loci (cpSSR) and variation at one maternally inherited mitochondrial marker (nad4‐3/4). Finally a subset of 11 populations was analysed with amplified fragment length polymorphism (AFLP). Comparisons were made with already available Abies sibirica data. For all sets of markers, we estimated genetic diversity and differentiation using an analysis of molecular variance (AMOVA). Population clustering was assessed with a Bayesian approach implemented in structure v.2.3. Results Among the three major species, A. sibirica, A. nephrolepis and A. sachalinensis, A. sachalinensis demonstrated the highest cytoplasmic and nuclear diversity and the most continental species, A. sibirica, the lowest. Both nuclear and mitochondrial DNA markers revealed the presence of a transitional zone on Sakhalin Island between A. nephrolepis and A. sachalinensis of south Sakhalin. The structure analysis delivered very clear results confirming the admixed origin of A. sachalinensis, with a genetic contribution from A. nephrolepis. No variation in cytoplasmic markers was found in A. gracilis, suggesting the occurrence of a recent bottleneck. Main conclusions There is a clear reduction of genetic diversity in Abies species from the Pacific coast into the continent. The higher diversity in A. sachalinensis could have two causes: a larger effective population size in the islands due to relatively stable climatic conditions and consequently less pronounced demographic fluctuations in population size and/or hybridization with continental and Japanese populations. Sakhalin Island is a major transitional zone for conifer species. Finally, the fir from Kamchatka, A. gracilis, should be regarded as a separate species closely related to the A. nephrolepis–A. sachalinensis complex.     

Invasiveness Does Not Predict Impact: Response of Native Land Snail Communities to Plant Invasions in Riparian Habitats

Studies of plant invasions rarely address impacts on molluscs. By comparing pairs of invaded and corresponding uninvaded plots in 96 sites in floodplain forests, we examined effects of four invasive alien plants (Impatiens glandulifera, Fallopia japonica, F. sachalinensis, and F.×bohemica) in the Czech Republic on communities of land snails. The richness and abundance of living land snail species were recorded separately for all species, rare species listed on the national Red List, and small species with shell size below 5 mm. The significant impacts ranged from 16–48% reduction in snail species numbers, and 29–90% reduction in abundance. Small species were especially prone to reduction in species richness by all four invasive plant taxa. Rare snails were also negatively impacted by all plant invaders, both in terms of species richness or abundance. Overall, the impacts on snails were invader-specific, differing among plant taxa. The strong effect of I. glandulifera could be related to the post-invasion decrease in abundance of tall nitrophilous native plant species that are a nutrient-rich food source for snails in riparian habitats. Fallopia sachalinensis had the strongest negative impact of the three knotweeds, which reflects differences in their canopy structure, microhabitat humidity and litter decomposition. The ranking of Fallopia taxa according to the strength of impacts on snail communities differs from ranking by their invasiveness, known from previous studies. This indicates that invasiveness does not simply translate to impacts of invasion and needs to be borne in mind by conservation and management authorities.     

Genetic,Epigenetic, and HPLC Fingerprint Differentiation between Natural and Ex Situ Populations of Rhodiola sachalinensis from Changbai Mountain,China

Rhodiola sachalinensis is an endangered species with important medicinal value. We used inter-simple sequence repeat (ISSR) and methylation-sensitive amplified polymorphism (MSAP) markers to analyze genetic and epigenetic differentiation in different populations of R. sachalinensis, including three natural populations and an ex situ population. Chromatographic fingerprint was used to reveal HPLC fingerprint differentiation. According to our results, the ex situ population of R. sachalinensis has higher level genetic diversity and greater HPLC fingerprint variation than natural populations, but shows lower epigenetic diversity. Most genetic variation (54.88%) was found to be distributed within populations, and epigenetic variation was primarily distributed among populations (63.87%). UPGMA cluster analysis of ISSR and MSAP data showed identical results, with individuals from each given population grouping together. The results of UPGMA cluster analysis of HPLC fingerprint patterns was significantly different from results obtained from ISSR and MSAP data. Correlation analysis revealed close relationships among altitude, genetic structure, epigenetic structure, and HPLC fingerprint patterns (R² = 0.98 for genetic and epigenetic distance; R² = 0.90 for DNA methylation level and altitude; R² = –0.95 for HPLC fingerprint and altitude). Taken together, our results indicate that ex situ population of R. sachalinensis show significantly different genetic and epigenetic population structures and HPLC fingerprint patterns. Along with other potential explanations, these findings suggest that the ex situ environmental factors caused by different altitude play an important role in keeping hereditary characteristic of R. sachalinensis.