Zhiqiang Han

Genetic population structure of Liza haematocheilus in north-western Pacific detected by amplified fragment length polymorphism markers

Several divergent sympatry mtDNA lineages have been described in redlip mullet Liza haematocheilus , and this high inter-lineage divergence raises questions about the taxonomic status of L. haematocheilus lineages in the north-western Pacific. In this study, the amplified fragment length polymorphism technique was employed to examine genetic structure of L. haematocheilus and estimate the level of independence of the different mtDNA lineages in the north-western Pacific. A total of 186 bands were amplified from 91 individuals among 8 populations by 4 primer combinations and the percentage of polymorphic bands was 91.74%. The Unweighted Pair Group Method with Arithmetic Mean tree based on Nei genetic distance revealed two clusters (North Clade and South Clade). Molecular variance analysis and pairwise F ST supported the separation of north and south populations of L. haematocheilus in the north-western Pacific. The incongruence between nuclear groups and mitochondrial lineages suggests the three distinct lineages do not represent cryptic species and the presence of divergent mitochondrial lineages in the same sample is a result of secondary contact after an extended period of isolation. The Pleistocene isolation and biological characteristics of species may be responsible for the genetic differentiation of L. haematocheilus .

Genomic evidence for local adaptation in the ovoviviparous marine fish Sebastiscus marmoratus with a background of population homogeneity

Advances in next-generation sequencing techniques have allowed for the generation of genome-wide sequence data, to gain insight into the dynamics influencing genetic structure and the local adaptation of marine fish. Here, using genotyping-by-sequencing (GBS) technique, we identified 31,119 single nucleotide polymorphisms (SNPs) for Sebastiscus marmoratus in 59 individuals from three populations in Chinese coastal waters. Based on all SNPs, there was little evidence of genetic differentiation among populations. However, outlier tests revealed 329 SNPs putatively under divergent selection across populations. Structural and phylogenetic topology analyses based on the outliers showed clear genetic differentiation among populations. Gene Ontology (GO) annotation results revealed that most of these outliers are known or hypothesized to be involved in metabolic process. Together with previous work using mitochondrial cytochrome b sequences, the present results further suggest that the population structure is strongly influenced by locally adaptive pressure. Overall, adaptive evolution in a heterogeneous environment plays an important role in inducing genetic differentiation among local populations. This study increases understanding of the factors (including gene flow and local adaptation) promoting and constraining population genetic differentiation in marine organisms.

The Possible Physical Barrier and Coastal Dispersal Strategy for Japanese Grenadier Anchovy, Coilia nasus in the East China Sea and Yellow Sea: Evidence from AFLP Markers

In order to ascertain the taxonomic status of the Ariake Sea population of Japanese grenadier anchovy, Coilia nasus, and assess the contemporary possible genetic barrier between the west and east coastal waters of the East China Sea, we used amplified fragment length polymorphism (AFLP) markers to detect the genetic structure of C. nasus, in the East China Sea and Yellow Sea. Eighty-one individuals of C. nasus were collected from five locations and 12 individuals of Coilia mystus were sampled from the Yangtze River Estuary. A total of 371 loci were detected by five primer combinations, 310 of which were polymorphic (83.56%). Analysis of molecular variation (AMOVA) and pairwise fixation index (FST) revealed significant genetic differentiation among five samples, indicating limited gene flow among populations. The dendrogram for populations by neighbor-joining (NJ) cluster analysis provided evidence of a clear relationship between genetic and geographic patterns, supporting significant genetic differentiation between China coastal populations and Ariake Sea populations. Compared to the genetic divergence between C. nasus and C. mystus, the level of genetic differentiation between China and the Ariake Sea populations of C. nasus is obvious below the species level, indicating isolated populations of C. nasus in the Ariake Sea. Isolation by distance analysis revealed that direct ocean distance with deep-water at the continental slope and high salinity between west and east coastal waters of the East China Sea served as major physical barrier to C. nasus, supporting the coastal dispersal pattern in this estuarine species, and rejecting offshore dispersal strategy.

Is the Kuroshio Current a strong barrier for the dispersal of the gizzard shad (Konosirus punctatus) in the East China Sea

The gizzard shad (Konosirus punctatus) is a common pelagic fish in the East China Sea. To evaluate the influence of Kuroshio Current in shaping the genetic structure of marine species, 10 populations of K. punctatus from Chinese and Japanese coastal waters were collected for analysis based on the mitochondrial DNA marker. All the Japanese populations exhibited higher nucleotide diversity than did Chinese populations. Two distinct clades were identified by the neighbour-joining tree based on haplotypes of Cyt b and the control region. Both AMOVA and pairwise Fst strongly supported the significant genetic divergence between Chinese and Japanese clades, which suggested strictly limited gene exchange. The mismatch distribution of Chinese clade and Japanese Clade B appeared to be unimodal, and Tajima’s D and Fu’s Fs statistics were significantly negative, and, therefore, the hypothesis of selective neutrality was rejected. The results indicated that the Kuroshio Current may not be the strong barrier for the dispersal of K. punctatus in the East China Sea. The climate of Pleistocene periods had played an important role in phylogeographical patterns of K. punctatus and the dispersal strategy of coastal species may be the major current physical barrier for the gene flow among populations from Chinese and Japanese coastal waters.

Genetic structure and unique origin of the introduced blue mussel Mytilus galloprovincialis in the north-western Pacific: clues from mitochondrial cytochrome c oxidase I (COI) sequences

As an introduced species, the Mediterranean blue mussel Mytilus galloprovincialis is widely distributed in the north-western Pacific. However, the mussels introduction pathway, colonisation route and source of introduction are unclear. In the present study, genetic variation among 11 colonised populations of M. galloprovincialis in the north-western Pacific was analysed using fragments of the mitochondrial cytochrome c oxidase I (COI) gene. Analysis of molecular variance (AMOVA) detected no significant differences at any hierarchical level, and all fixation index (FST) values were non-significant, reflecting a founder event and potential high dispersal ability. Phylogenetic analysis showed that populations of M. galloprovincialis in the north-western Pacific were introduced from the middle Mediterranean Sea and that the Dalian population was the first colonised population in the coastal waters of China. By assessing the origin of introduced populations of M. galloprovincialis, the present study provides new insights into the biogeography of M. galloprovincialis.

Morphological characteristics and DNA barcoding of Pampus echinogaster (Basilewsky, 1855)

The morphological similarities of Pampus fishes have led to considerable confusion in species-level identification, and no accurate information on neotype or DNA barcoding of Pampus echinogaster is available. Two hundred and seven specimens of P. echinogaster were collected from the coastal waters of Dandong, Dongying, Qingdao, Nantong, Zhoushan, Wenzhou, Changle, Taiwan, and Wakayama (Japan), from June 2010 to April 2013. The diagnostic characteristics of P. echinogaster are as follows: dorsal fin VIII-XI-43–51, anal fin V-VIII-43–49, pectoral fin 22–27, caudal fin 19–22, pelvic fin absent; first gill rakers sparse, slender (pointed), 3–4+12–16=15–20; vertebrae 39–41; transverse occipital canal on top of head moderately small, wavy ridges not reaching upper origin of pectoral fin; ventral branch of lateral line canal spare, shorter than dorsal branch of lateral line canal. By combining congener sequences of the cytochrome oxidase I (COI) gene from GenBank, two absolute groups were detected among all specimens, which further indicated that two valid species were present based on genetic differences in amino acid sequences and the distance between the groups. The sequences of Group 1 can be regarded as DNA barcoding of P. echinogaster. The correct morphological redescription and DNA barcoding of P. echinogaster are presented here to provide a guarantee for efficient and accurate studies, a theoretical basis for classification, and enable appropriate fishery management and conservation strategies for the genus Pampus in the future.

Genetic structure analysis of mantis shrimp Oratosquilla oratoria based on mitochondrial DNA control region sequence

The genetic relationships of mantis shrimp Oratosquilla oratoria between the coastal waters of China and Japan were not well studied. To reveal the genetic differentiation and genetic structure among populations, we collected populations of mantis shrimp O. oratoria from the coastal waters of China and Japan to analyze the mtDNA control region variation. A total of 309 individuals of O. oratoria were collected from 13 localities (11 from China and 2 from Japan) and a segment of mitochondrial DNA control region was sequenced. Three hundred nine haplotypes were defined, yielding a very high haplotype diversity and nucleotide diversity. Two lineages of O. oratoria were revealed and displayed strong differences in the geographical distribution. In the coastal waters of China, the geographic distribution of the two lineages was completely separated by the Yangtze River estuary; however, the lineages showed geographic sympatry in two populations from Japan. Based on the lineage distribution, three groups were defined. There was no significant genetic differentiation among the populations within the three groups, indicating high gene flow within each group. Significant and negative values for Tajima D and Fu’s Fs tests, and mismatch distributions for two lineages indicated population expansion. The present result confirmed that the freshwater outflow from the Yangtze River formed a physical barrier and affected gene exchange. The different distribution patterns of the two lineages in coastal waters of China and Japan indicated that the larvae of O. oratoria were transferred from China to the coastal waters of Japan with a one-way gene flow.

Morphological characters and DNA barcoding of Syngnathus schlegeli in the coastal waters of China

A Syngnathus species widely distributed in Chinese seas was permanently identified as Syngnathus acus by native ichthyologists, but the taxonomic description about this species was inadequate and lacking conclusively molecular evidence. To identify this species, 357 individuals of this species from the coastal waters of Dandong, Yantai, Qingdao and Zhoushan were collected and measured. Morphological results showed that these slender specimens were mainly brownish, usually mottled with pale. Standard length ranged from 117 mm to 213 mm with an average length of 180.3 mm. The above characters were consistent with S. schlegeli distributed in Japan but colored differently from and much smaller than typical S. acus reported in Europe. Thus, morphological studies revealed that this species was previously misidentified as S. acus and might be S. schlegeli in reality. In addition, a fragment of cytochrome oxidase subunit I (COI) gene of mitochondrial DNA was also sequenced for species identification, and 15 COI sequences belonging to different Syngnathus species were also used for the molecular identification. COI sequences of our specimens had the minimum genetic distance from recognized S. schlegeli from Japan and clustered with it firstly. The phylogenetic analysis similarly suggested that the species previously identified as S. acus in the coastal waters of China was S. schlegeli actually.

Restricted Gene Flow for Gadus macrocephalus from Yellow Sea Based on Microsatellite Markers: Geographic Block of Tsushima Current

The Pacific cod Gadus macrocephalus is a demersal, economically important fish in the family Gadidae. Population genetic differentiation of Pacific cod was examined across its northwestern Pacific range by screening variation of eight microsatellite loci in the present study. All four populations exhibited high genetic diversity. Pairwise fixation index (Fst) suggested a moderate to high level of genetic differentiation among populations. Population of the Yellow Sea (YS) showed higher genetic difference compared to the other three populations based on the results of pairwise Fst, three-dimensional factorial correspondence analysis (3D-FCA) and STRUCTURE, which implied restricted gene flow among them. Wilcoxon signed rank tests suggested no significant heterozygosity excess and no recent genetic bottleneck events were detected. Microsatellite DNA is an effective molecular marker for detecting the phylogeographic pattern of Pacific cod, and these Pacific cod populations should be three management units.

Effects of Pleistocene glaciation on the phylogeographic and demographic histories of chub mackerel Scomber japonicus in the north-western Pacific

Pleistocene glacial cycles associated with geological and climatic changes have been suggested to affect the distribution and abundance of marine organisms in the north-western Pacific. In addition to historical processes, several contemporary forces are associated with spatial distributions and genetic structuring of marine species. Herein, we gathered partial mitochondrial control region sequences of chub mackerel Scomber japonicus from 14 localities over most of its geographical range in the north-western Pacific to infer the effect of Pleistocene climatic fluctuations on its historical demography, and to assess the role of historical process and contemporary factors in shaping present-day patterns of genetic differentiation within this species. Phylogeographic patterns revealed two distinct lineages that originated in vicariance during the Middle Pleistocene. However, there was no evidence of phylogeographic partitioning of haplotypes over the sampled range. Population structure analyses indicated a high degree of genetic homogeneity among chub mackerel populations. Demographic analyses indicated both lineages experienced Late Pleistocene population expansion. The observed genetic pattern of chub mackerel is consistent with a scenario of the survival of this species in at least two allopatric glacial refugia during the glacial maxima of the Pleistocene followed by massive dispersals throughout the north-western Pacific and species-specific ecological processes facilitating contemporary gene flow.