Desiccation resistance of two semiterrestrial isopods, Ligia exotica and Ligia taiwanensis (Crustacea) in Taiwan

The ability to resist desiccation stress was examined in two semiterrestrial Ligia species, Ligia exotica Roux and L. taiwanensis Lee, in Taiwan, under a certain desiccation condition. L. exotica exhibited the longer survival time, lower weight-specific rates of water loss, and the slightly higher ability of tolerance to water loss, compared to L. taiwanensis. In each species, the animal size displays a positive correlation to the survival time and total ability to resist desiccation, yet this size effects on the weight-specific water loss rate is negative. Neither water content nor maximum tolerance to water loss shows the association with the animal size in both species. The path ways and magnitudes of the interactions between these traits of desiccation resistance are analyzed and diagrammed using a stepwise regression model. In this model, the body sizes of animal can explain the most part of the variations in the survival time. The body size has a direct effect and an indirect effect, through the effect on water loss rate, on the time that the experimental animals can survival under this desiccated condition. These results suggest that L. exotica attains larger size than does L. taiwanensis, a lower transpiration rate and, consequently, a greater ability in desiccation resistance. The performances of these interactions in the desiccated resistance are more advantageous for L. exotica to migrate and colonize in variable land habitats within a certain limit, and as a result that L. exotica shows a broader distribution pattern than did L. taiwanensis in Taiwan.     

Microsatellite DNA analysis reveals lower than expected genetic diversity in the threatened leopard cat (<Emphasis Type="Italic">Prionailurus bengalensis</Emphasis>) in South Korea

To optimize conservation efforts, it is necessary to determine the risk of extinction by collecting reliable population information for a given species. We developed eight novel, polymorphic microsatellite markers and used these markers in conjunction with twelve existing markers to measure genetic diversity of South Korean populations of leopard cat (Prionailurus bengalensis), a species for which population size and habitat area data are unknown in the country, to assess its conservation status. The average number of alleles and the observed heterozygosity of the species were 3.8 and 0.41, respectively, and microsatellite diversity was lower than the average genetic diversity of 57 populations of 12 other felid species, and lower than that of other mammal populations occurring in South Korea, including the raccoon dog (Nyctereutes procyonoides), water deer (Hydropotes inermis), and endangered long-tailed goral (Naemorhedus caudatus). Furthermore, analysis of genetic structure in the national leopard cat population showed no clear genetic differentiation, suggesting that it is not necessary to divide the South Korean leopard cat population into multiple management units for the purposes of conservation. These results indicate that the genetic diversity of the leopard cat in South Korea is unexpectedly low, and that the risk of local extinction is, as a result, substantial. Thus, it is necessary to begin appropriate conservation efforts at a national level to conserve the leopard cat population in South Korea.     

New occurrence of the invasive alien leaf‐footed bug Leptoglossus gonagra (Hemiptera: Coreidae) in South Korea

The invasive alien leaf‐footed bug Leptoglossus gonagra (Fabricius 1775), native to the Americas, is herein recorded from South Korea for the first time. Forty nymphs and adults were collected on 22 September 2016 from arable lands of two separate sites. These individuals were identified based on the analysis of morphological characters and cytochrome oxidase c subunit 1 sequences. A neighbor‐joining tree indicated the existence of two clades: Group A from Brazil, and Group B from Asia and Oceania. Mean genetic distance between clades was 8.11%, range 7.60–8.57%. Within clades, mean genetic distance was 0.42% (0.17–0.83%) in Group A and, 0.45% (0.00–0.83%) in Group B. Colonies of 2nd‐final instar nymphs and adults were found, on top of leaves, flowers, fruits, and between the stems of pumpkins (Cucurbita moschata). Nymphs and adults damaged host plants by sucking their leaves, stems, fruits, and buds. After the first discovery in 2016, we were unable to find the bugs again (in 2017–2019). Therefore, the invasion of this species seems to have occurred only temporarily, during the warm season. We suggest that the species may have failed to overwinter as it is normally distributed in regions warmer than Korea. Nevertheless, possible appearances of this species must be continuously monitored as it is a potential pest of various plants cultivated in South Korea.     

Landscape-level spatial genetic structure in Quercus acutissima (Fagaceae)

Quercus acutissima (Fagaceae), a deciduous broad-leaved tree, is an important forest element in hillsides of South Korea. We used allozyme loci, Wright's F statistics, and multilocus spatial autocorrelation statistics to examine the distribution of genetic diversity within and among three local populations and the spatial genetic structure at a landscape scale (15 ha, 250 × 600 m) on Oenaro Island, South Korea. Levels of genetic diversity in Q. acutissima populations were comparable to mean values for other oak species. A moderate but significant deficit of heterozygotes (mean F(IS) = 0.069) was detected within local populations and low but significant differentiation was observed among populations (F(ST) = 0.010). Spatial autocorrelation analyses revealed little evidence of significant genetic structure at spatial scales of 100-120 m. The failure to detect genetic structure within populations may be due to intraspecific competition or random mortality among saplings, resulting in extensive thinning within maternal half-sib groups. Alternatively, low genetic differentiation at the landscape scale indicates substantial gene flow among local populations. Although wind-borne pollen may be the primary source of gene flow in Q. acutissima, these results suggest that acorn movement by animals may be more extensive than previously anticipated. Comparison of these genetic data for Oenaro Island with a disturbed isolated inland population suggests that population-to-population differences in internal genetic structure may be influenced by local variation in regeneration environment (e.g., disturbance).     

Morphological and molecular data confirm species assignment and dispersal of the genus Ligia (Crustacea: Isopoda: Ligiidae) along northeastern coastal China and East Asia

The morphological traits and mitochondrial genes of sympatric sea slaters from the genus Ligia were analysed from samples collected along northeastern coastal China and adjacent coastal East Asia to assign the species and to obtain interpretations of their distributions. Using 338 sequences and 199 diagnoses of 16S polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) from 331 specimens sampled from 26 localities, and homological sequences from counterparts from South Korea and the west coast of the USA, we assigned these taxa into two distinct species, Ligia exotica and Ligia cinerascens. Both species are recovered as two monophyletic clades in the robust phylogenetic hypothesis presented here. The northern boundaries of these species in China and the Korean peninsula are more southern than those of counterparts in Japan. Ligia exotica in northeastern coastal China is further split into two geographical subclades (B1 and B2), coexisting near 35°N. The distribution range of L. cinerascens covers the whole range of L. exotica subclade B1. Counterparts in Japan show a similar pattern. Oceanic rafting, shipping, damming and planting of seaweed are proposed as the media for the contemporary connectivity among populations of Ligia species inhabiting coastal China. © 2013 The Linnean Society of London     

Multiplication of Chilo iridescent virus in noninsect arthropods

Attempts to infect noninsect arthropods with Chilo iridescent virus (CIV) originally isolated from Lepidoptera were made by using eight species belonging to four classes. Multiplication of CIV was demonstrated in two species of terrestrial Crustacea (the pill bug, Armadillidium vulgare, and the slater, Porcello scaber) and one species of Chilopoda, the house centipede, Thereuonema higendorfi. The lethality experiment of CIV for A. vulgare suggested that chronic infection is a characteristic of the CIV infection in both classes, Crustacea and Insecta. Neither iridescence nor recovery of virus infectivity was demonstrated in the following arthropod species: the sea slater, Ligia exotica (Crustacea: Isopoda), the grapsid crab, Sesarma haematocheir (Crustacea: Decapoda), the millipede, Oxidus gracillis (Diplopoda: Polydesmoidea), Rhysodesmus semicirculatus (Diplopoda: Polydesmoidea), and the giant crab spider, Heteropoda venatoria (Arachnida: Araneae).     

Genetic diversity and genetic structure of the endangered Manchurian trout, <Emphasis Type="Italic">Brachymystax lenok tsinlingensis</Emphasis>, at its southern range margin: conservation implications for future restoration

The Manchurian trout, Brachymystax lenok tsinlingensis (family: Salmonidae), is a cold freshwater fish endemic to Northeast Asia. South Korean populations of this species, which comprise its southern range limit, have recently decreased markedly in size and are now becoming critically endangered. We assessed the current population status of this species in South Korea by estimating the levels of genetic diversity and genetic structure of five natural and four restored populations using mitochondrial DNA (mtDNA) control region sequences and eight nuclear microsatellite loci. Levels of within-population genetic diversity were low, suggesting that past effective population sizes (N _e) have been small. Each population had one or a maximum of two mtDNA haplotypes. Microsatellite allelic richness (AR) was significantly higher for natural populations (mean AR?=?3.51; 95% confidence interval, 3.00–4.03) than for restored populations (mean AR?=?2.61; 2.38–2.98). South Korean populations were significantly genetically isolated from one another, with private mtDNA haplotypes and microsatellite alleles, suggesting that limited gene flow has been occurring among populations. A mtDNA phylogeny revealed that South Korean lineages were more closely related to those of China than to those of North Korea and Russia. Overall, we suggest that future restoration efforts aimed at South Korean populations should consider the genetic characteristics reported here, which should help to fulfil effective conservation strategies for this highly cherished species. Our results will inform other conservation efforts, including assisted migration of freshwater fish populations at the equatorial end of the geographical range limit of the species.     

基于mtDNA-COI基因序列的雷氏按蚊分子群体遗传结构研究

【目的】探讨我国雷氏按蚊Anopheles lesteri的群体差异和分化程度。【方法】采用PCR方法, 从分子水平鉴别了采自我国和韩国的雷氏按蚊共9个群体139个样本, 扩增其线粒体DNA细胞色素氧化酶亚基Ⅰ基因, 并进行序列测定和分析。【结果】本研究共获得49个单倍型, 各单倍型呈高水平的平行演化, 来自云南群体的单倍型显示是扩张的源头。分子变异等级分析(AMOVA)的计算结果显示, 群体内变异占总变异的比例（64.95%）大于群体间（35.05%）, FST值为0.3504, 各群体间出现遗传分化。Mantel检验结果显示基因流水平与地理距离呈负相关关系（R²＝0.1322）, 群体遗传结构符合距离隔离模型。【结论】雷氏按蚊韩国和辽宁群体与其他分布地群体间差异大, 已出现明显分化, 我国其他分布地群体间的遗传差异小。     

Genetic diversity and population structure of the Korean field mouse(Apodumus peninsulae) in South Korea: from 17 previously and newly developed microsatellite markers

To provide more reliable genetic information on species and minimize experimental errors, biologists increase the number of genetic markers available and then carefully select optimal markers from a large candidate pool. We developed nine novel microsatellite markers from the Korean field mouse (Apodemus peninsulae), which is one of the most dominant forest animals in South Korea. The mean observed and expected heterozygosities across nine markers were 0.65 and 0.73, respectively, with an average polymorphic information content of 0.70. Using 17 microsatellite markers (nine polymorphic markers in this study, in combination with eight previously reported for the species), we conducted genetic analysis on the animals from six sampling locations. These locations are divided into the eastern (EAST) and the western (WEST) sides of the Taebaek mountain ranges in South Korea. Genetic diversity was high at both groups, with the mean expected heterozygosity of 0.77 in EAST and 0.78 in WEST. However, we did not observe strong evidence of genetic divergence between two groups. Future genetic research with more samples incorporating ecological study may clarify population structure in the species and the hypothesis of the mountains discontinuity of gene flow.     

Genetic differences between wild and hatchery populations of red sea cucumber (Stichopus japonicus) inferred from microsatellite markers: implications for production and stocking programs design

Red sea cucumber Stichopus japonicus is the most important and valuable commercial sea cucumber species in Korea. Its farming and stock enhancement started in the early 2000s and is still rapid expansion in Korea. Therefore, the analyses of genetic status of wild and hatchery populations are necessary to maintain the genetic diversity of this valuable marine resource. In this study, possible genetic similarity and differences between the wild population and hatchery population in Jeju, Korea were accessed using multiplex assays with eight highly polymorphic microsatellite loci. High levels of polymorphism were observed between the two populations. A total of 93 different alleles were found. Although a considerable loss of unique alleles and relatively high inbreeding coefficient value were observed in the hatchery samples, no statistically significant reductions were found in heterozygosity or allelic diversity in the hatchery population, compared with the wild population. However, significant genetic heterogeneity was found between two populations. These results suggest that genetic drift has probably promoted differentiation between populations, and stocking intensity in wild populations may correlate with loss of genetic integrity. Therefore, the sustainable exploitation plans of the fishery resource should be developed by applying basic genetic principles combined with molecular monitoring. This genetic baseline information of Korean red sea cucumber has important implications for designing of genetically sustainable restocking programs and more efficient conservation management guidelines for these valuable genetic materials.     

Weak large‐scale population genetic structure in a philopatric seabird,the European Shag Phalacrocorax aristotelis

Quantifying population genetic structure is fundamental to testing hypotheses regarding gene flow, population divergence and dynamics across large spatial scales. In species with highly mobile life‐history stages, where it is unclear whether such movements translate into effective dispersal among discrete philopatric breeding populations, this approach can be particularly effective. We used seven nuclear microsatellite loci and mitochondrial DNA (ND2) markers to quantify population genetic structure and variation across 20 populations (447 individuals) of one such species, the European Shag, spanning a large geographical range. Despite high breeding philopatry, rare cross‐sea movements and recognized subspecies, population genetic structure was weak across both microsatellites and mitochondrial markers. Furthermore, although isolation‐by‐distance was detected, microsatellite variation provided no evidence that open sea formed a complete barrier to effective dispersal. These data suggest that occasional long‐distance, cross‐sea movements translate into gene flow across a large spatial scale. Historical factors may also have shaped contemporary genetic structure: cluster analyses of microsatellite data identified three groups, comprising colonies at southern, mid‐ and northern latitudes, and similar structure was observed at mitochondrial loci. Only one private mitochondrial haplotype was found among subspecies, suggesting that this current taxonomic subdivision may not be mirrored by genetic isolation.     

Microsatellite variation in the pinewood nematode, Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle in South Korea

The pinewood nematode Bursaphelenchus xylophilus is the causative agent of pine wilt disease, which has caused heavy economic losses to the South Korean forest industry. In this study, we investigated the genetic variation among South Korean pinewood nematodes using newly developed microsatellite loci. In order to ensure sufficient templates for the amplification of multiple loci required for individual identification, we employed an amplifying step of restricted fragments during the microsatellite development procedure. We found atypical genetic patterns in this non-native pest species: high allelic diversity and population structure. The large number of alleles may be the result of continuous and/or large-scaled introduction, which apparently went unnoticed before the first official report of pine wilt disease in Korea in 1998, or may come from gene pools of closely related species through genetic introgression after hybridization. Ecological properties of this species, such as a vector-mediated life cycle, may have contributed to its population structure, which may be enhanced by governmental efforts to prevent dispersal of this disease. As a geographic population structure was not observed, geographic patterns of genetic variation appear to be more affected by anthropogenic mediation than by natural dispersion through vector insects. And genotypes of Korean populations were compared to genotypes found in neighboring countries such as China and Japan.     

Effects of topography on dispersal of black-billed magpie Pica pica sericea revealed by population genetic analysis

This study was conducted using random amplified polymorphic DNA analysis to examine whether the effects of topographical structure on the dispersal of black-billed magpie Pica pica sericea are reflected in its population genetic structure. The black-billed magpie is typically seen in the lowlands and is remarkably sedentary in Korea and Japan. The unweighted pair-group method of clustering analysis showed two main clusters: five populations in the western region of the Bekdudegan mountains (WRBM) in one cluster and five populations in the eastern region of the Bekdudegan mountains (ERBM), including the Japanese population in the other cluster. The populations in WRBM appeared to be diverged from the populations in ERBM. Analysis of molecular variance revealed that the populations in ERBM were more genetically divergent from each other than were those in WRBM. The high-rising mountains, very rugged topographical features and the sea in ERBM may have resulted in a lower dispersal rate and larger genetic variation among populations in ERBM compared to those in WRBM. These results suggest that the topographical structure may have an influence on the dispersal and population genetic structure of the black-billed magpie. Electronic Publication     

Distribution and genetic characteristics of Ephoron shigae (Ephemeroptera: Polymitarcyidae) in Korea

Ephoron shigae is distributed in Far East Russia, Korea and Japan, where it inhabits rivers, streams and lakes. Although this mayfly is a geographically parthenogenetic species, unisex populations have been observed only in Japan. We aimed to answer the following three questions: (i) whether there is a unisex population of E. shigae in Korea; (ii) whether migration and gene flow exists between E. shigae populations in Korea and Japan; and (3) whether E. shigae populations are fragmented in Korea because of mountains running to the north and south. We observed 20 populations across Korea and investigated the sex ratio in 12 populations. Our results showed that all the populations were bisex, and we believe that there is no unisex population of E. shigae in Korea. In addition, we examined the genetic structure of E. shigae in Korea by using the COI gene. The results showed that the Korean populations were clearly divergent from the Japanese populations; the level of genetic distance between the Korean and Japanese populations was interspecific rather than intraspecific. Therefore, parthenogenetic populations of E. shigae probably belong to unisex populations that originated in Japan, while the individuals would have not migrated to Korea. The Korean populations did not show clear fragmentation by geographic barrier, although this mayfly with extremely short adult stage (up to 2 h) could be a comparatively strong disperser across Korea.     

Population genetic structure,introgression, and hybridization in the genus Rhizophora along the Brazilian coast

Mangrove plants comprise plants with similar ecological features that have enabled them to adapt to life between the sea and the land. Within a geographic region, different mangrove species share not only similar adaptations but also similar genetic structure patterns. Along the eastern coast of South America, there is a subdivision between the populations north and south of the continent's northeastern extremity. Here, we aimed to test for this north‐south genetic structure in Rhizophora mangle, a dominant mangrove plant in the Western Hemisphere. Additionally, we aimed to study the relationships between R. mangle, R. racemosa, and R. × harrisonii and to test for evidence of hybridization and introgression. Our results confirmed the north‐south genetic structure pattern in R. mangle and revealed a less abrupt genetic break in the northern population than those observed in Avicennia species, another dominant and widespread mangrove genus in the Western Hemisphere. These results are consistent with the role of oceanic currents influencing sea‐dispersed plants and differences between Avicennia and Rhizophora propagules in longevity and establishment time. We also observed that introgression and hybridization are relevant biological processes in the northeastern coast of South America and that they are likely asymmetric toward R. mangle, suggesting that adaptation might be a process maintaining this hybrid zone.     

Isolation and characterization of eight microsatellite loci from <Emphasis Type="Italic">Lycorma delicatula</Emphasis> (White) (Hemiptera: Fulgoridae) for population genetic analysis in Korea

Lycorma delicatula (White) is native to China but is becoming an important insect pest in Korea. Polymorphic DNA markers like microsatellites are widely used for characterizing dispersal patterns and capacity of invasive insect pests which can contribute to designing effective management of the species. To facilitate such population genetic studies of L. delicatula in Korea, we isolated and characterized eight microsatellite loci for L. delicatula using a hybridization-biotin enrichment method. We further used these novel microsatellite loci to determine population genetic parameters for 33 L. delicatula specimens collected from Cheonan, South Korea where outbreaks of this species were first reported in Korea. The number of alleles per locus ranged from three to ten, with an average of 6.25. The mean expected (H _E) and observed heterozygosities (H _O) were 0.575 and 0.626, respectively. The eight loci showed no deviation from Hardy–Weinberg equilibrium according to the adjusted significance threshold (P = 0.00625), and there was no linkage disequilibrium between each pair of these eight markers. Bayesian cluster analysis using the program structure revealed no evidence of genetic structuring in L. delicatula samples from Cheonan. These new microsatellite markers will be widely applicable to future ecological genetic studies of L. delicatula, including assessment of the level of gene flow and genetic connectivity among populations that are necessary for effective management and monitoring of the species.     

A genetic comparison of Brucella abortus isolates from animals and humans by using an MLVA assay

The MLVA assay is known to have a high ability to identify and discriminate Brucella species, so that it can be used as an epidemiological tool to discriminate Brucella isolates originating from restricted geographic sources. In this study, the genetic profiles of 38 B. abortus isolates from humans were analyzed and compared with genotypes from animal isolates in South Korea. As a result, it was found that they did not show high genetic diversity and were compacted. They were clustered together with animal isolates, showing a significant correlation to regional distributions. With its ability to prove a significant genetic correlation among B. abortus isolates from animals and humans in South Korea, the MLVA assay could be utilized as part of a program to control and eradicate brucellosis, one of the major zoonoses. This study represents the first data of genetic correlation of B. abortus isolates from humans and animals in South Korea.     

Developmental changes in dopamine modulation of the heart in the isopod crustacean Ligia exotica: reversal of chronotropic effect

Developmental changes in dopamine modulation of the heart were examined in the isopod crustacean Ligia exotica. The Ligia cardiac pacemaker is transferred from the myocardium to the cardiac ganglion during juvenile development and the heartbeat changes from myogenic to neurogenic. In the myogenic heart of early juveniles, dopamine affected the myocardium and caused a decrease in the frequency and an increase in the duration of the myocardial action potential, resulting in negative chronotropic (decrease in beat frequency) and positive inotropic (increase in contractile force) effects on the heart. Contrastingly, in the heart of immature adults just after juvenile development, dopamine caused effects of adult type, positive chronotropic and positive inotropic effects on the heart affecting the cardiac ganglion and myocardium. During the middle and late juvenile stages, dopamine caused individually a negative or a positive chronotropic effect on the heart. These results suggest that the chronotropic effect of dopamine on the Ligia heart is reversed from negative to positive in association with the cardiac pacemaker transfer from the myocardium to the cardiac ganglion during juvenile development.     

Mitochondrial cytochrome b sequence variations and population structure of Siberian chipmunk (Tamias sibiricus) in Northeastern Asia and population substructure in South Korea

Twenty-five chipmunk species occur in the world, of which only the Siberian chipmunk, Tamias sibiricus, inhabits Asia. To investigate mitochondrial cytochrome b sequence variations and population structure of the Siberian chipmunk in northeastern Asia, we examined mitochondrial cytochrome b sequences (1140 bp) from 3 countries. Analyses of 41 individuals from South Korea and 33 individuals from Russia and northeast China resulted in 37 haplotypes and 27 haplotypes, respectively. There were no shared haplotypes between South Korea and Russia--northeast China. Phylogenetic trees and network analysis showed 2 major maternal lineages for haplotypes, referred to as the S and R lineages. Haplotype grouping in each cluster was nearly coincident with its geographic affinity. In particular, 3 distinct groups were found that mostly clustered in the northern, central and southern parts of South Korea. Nucleotide diversity of the S lineage was twice that of lineage R. The divergence between S and R lineages was estimated to be 2.98-0.98 Myr. During the ice age, there may have been at least 2 refuges in South Korea and Russia--northeast China. The sequence variation between the S and R lineages was 11.3% (K2P), which is indicative of specific recognition in rodents. These results suggest that T. sibiricus from South Korea could be considered a separate species. However, additional information, such as details of distribution, nuclear genes data or morphology, is required to strengthen this hypothesis.     

Characterization of 15 microsatellite loci and genetic analysis of Heterodera schachtii (Nematoda: Heteroderidae) in South Korea

Heterodera schachtii, the sugar beet cyst nematode, is a major pest of agricultural crops worldwide. We report the development of fifteen polymorphic microsatellite markers and assess the genetic diversity and structure of three populations following a recent invasion of a previously unaffected region. Populations had low levels of heterozygosity, likely indicative of population structure, history, and inbreeding. Genetic diversity analysis suggested that the current infestation in South Korea may have come either from a single source population of mixed ancestry, or from multiple sources, indicating that implementing adequate prevention measures is still an unmet challenge. Much more work is needed on this species to identify global patterns of spread, and the microsatellite loci we develop here should be useful in many regions for modeling range expansion, studying the evolution of resistance, and increasing the effectiveness of pest management strategies.