白鱼线粒体DNA控制区结构和种群遗传多样性分析

用特异性引物对白鱼(Anabarilius grahami)DNA进行PCR扩增,获得了白鱼线粒体DNA控制区基因全序列(930bp)。控制区T、C、A和G碱基组成为29.8%、22.5%、33.0%和14.7%。对照其他已报道的鱼类控制区结构,对白鱼控制区结构进行了分析,识别了其终止序列区、中央保守区和保守序列区,找到了终止相关的序列TAS以及保守序列(CSB-F、CSB-D、CSB-1、CSB-2、CSB-3)。同时运用DNA分析软件对白鱼一个驯养种群(中国科学院昆明动物研究所珍稀鱼类繁育中心)及两个自然地理种群(江川县明星鱼洞、江川县牛摩村)进行了遗传多样性分析。结果显示:两个自然种群存在较强基因交流,未出现遗传分化;人工驯养种群遗传多样性最高,种群复壮程度较好。     

白鱼线粒体DNA控制区结构和种群遗传多样性分析

用特异性引物对白鱼（Anabarilius grahami）DNA进行PCR扩增,获得了白鱼线粒体DNA控制区基因全序列（930 bp）。控制区T、C、A和G碱基组成为29.8%、22.5%、33.0%和14.7%。对照其他已报道的鱼类控制区结构,对白鱼控制区结构进行了分析,识别了其终止序列区、中央保守区和保守序列区,找到了终止相关的序列TAS以及保守序列（CSB-F、CSB-D、CSB-1、CSB-2、CSB-3）。同时运用DNA分析软件对白鱼一个驯养种群（中国科学院昆明动物研究所珍稀鱼类繁育中心）及两个自然地理种群（江川县明星鱼洞、江川县牛摩村）进行了遗传多样性分析。结果显示：两个自然种群存在较强基因交流,未出现遗传分化;人工驯养种群遗传多样性最高,种群复壮程度较好。     

松江鲈线粒体DNA控制区结构和遗传多样性分析

为探讨松江鲈(Trachidermus fasciatus Heckel)线粒体控制区特征及其群体遗传结构, 研究测定分析了中国和日本沿海共8个群体的线粒体控制区序列, 分析了其结构特征, 识别出终止序列区(ETAS)、中央保守区(CD)和保守序列区(CSB)的特征序列。遗传多样性分析结果显示: 69个松江鲈个体共检测到47个单倍型, 呈现出核苷酸多样性(0.0079)较低和单倍型多样性(0.978)较高的特点。单倍型邻接关系树和单倍型网络关系图均显示松江鲈分为中国和日本两大世系。遗传分化系数(Fst)和分子方差分析(AMOVA)结果表明, 松江鲈中国群体和日本群体之间存在的遗传差异较显著, 中国沿海各群体之间亦存在着一定程度的遗传差异, 该分化主要由历史环境变动、当代环境因素和自身生态习性等原因造成。     

长竹蛏不同地理居群的遗传多样性

研究以大连、烟台、莱州、青岛和赣榆近海5个不同地理居群的长竹蛏(Solen strictus Gould)为实验材料,利用ISSR分子标记进行了遗传多样性的研究。结果表明,13个ISSR引物在5个居群中共扩增出200个位点,平均每个引物记录15.4个位点,5个居群的多态位点比例为43.56%—60.43%。长竹蛏在物种水平上的Nei’s基因多样性指数和Shannon’s信息指数分别为0.2854和0.4390,在居群水平上分别为0.1674和0.2530。NJ聚类分析显示,青岛居群与赣榆居群的亲缘关系最近,而烟台居群与其他4个居群的亲缘关系较远。经Mantel检测,长竹蛏5个居群间的遗传距离与地理距离并无相关性(r=-0.0834,P>0.1)。AMOVA分子变异分析表明,长竹蛏的遗传变异有47.71%发生在居群间,52.29%发生在居群内,居群内的遗传变异大于居群间的遗传变异。长竹蛏5个居群间的遗传分化系数(Gst)为0.2889,基因流(Nm)为1.3194。结果表明,长竹蛏具有较高的遗传多样性,但居群间已发生了一定程度的遗传分化。       

南蝠线粒体DNA控制区结构及贵州7个种群遗传多样性的分析研究

采用PCR技术获得了贵州7个南蝠(Ia io)自然种群42个个体的线粒体DNA控制区全序列,长度为1256～1340 bp.对控制区结构进行分析,识别了其延伸的终止结合序列区(包括ETAS1和ETAS2元件)、中央保守区(包括F、E、D、C、B元件)和保守序列区(包括CSB1、CSB2和CSB3元件);同时,在延伸的终止结合序列区还发现了若干能形成发夹结构的主体序列TACAT—ATGTA.在7个自然种群42个个体中共定义了16个单倍型.遗传多样性分析表明:贵州南蝠种群具有较高的单倍型多样性(h=0.945)和中等的核苷酸多样性(π=0.012).基因流、AMOVA和系统进化树分析表明贵州这7个南蝠自然群体间没有发生遗传分化.     

Low mtDNA Cytb diversity and shallow population structure of Eleutheronema tetradactylum in the East China Sea and the South China Sea

Eleutheronema tetradactylum is an economically important fish species in China water. To investigate the genetic diversity and describe population structure of it, an 1151 base pair (bp) fragment of the mitochondrial DNA Cytb sequence was analyzed in 120 individuals from four populations in the East China Sea and the South China Sea. A total of 16 haplotypes were defined by 24 variable nucleotide sites. High level of haplotype diversity and low nucleotide diversity were observed in all populations. The results of AMOVA detected that 89.44% of the genetic variation occurred within populations. Significant genetic differentiations were detected among populations (0.05097, P < 0.05), but no large-scale regional differences were detected. Analysis of neutral evolution and mismatch distribution suggested no recent population expansion happened. The present results provided new information for genetic assessment, fishery management and conservation of this species.     

Genetic analysis of the populations of Japanese anchovy Engraulis japonicus from the Yellow Sea and East China Sea based on mitochondrial cytochrome b sequence

Japanese anchovy Engraulis japonicus was not only a commercial species but also a key link in the marine food chain in the Yellow Sea and East China Sea. However, in recent years, E. japonicus experienced a deep recession caused by over fishing. Therefore, determination of population genetic structure is essential to underpin resource recovery and to aid delineating and monitoring populations for fishery management. In this study, sequence variation of a 402-bp fragment of mitochondrial cytochrome b (cyt b) gene from ten populations of E. japonicus in the Yellow Sea and East China Sea was determined, detecting 73 haplotypes. Haplotype diversities and nucleotide diversities for the ten populations ranged from 0.4706 ± 0.1177 (D) to 0.9935 ± 0.210 (WH), and from 0.0014 ± 0.0013 (D) to 0.0090 ± 0.0053 (WH), respectively. Analysis of molecular variance (AMOVA) showed no significant differences at all hierarchical levels, and almost all F_ST values were non-significant, indicating no significant population genetic structure in the Yellow Sea and East China Sea. Two indistinct clades with low support values were detected and the analysis of haplotype frequency distribution showed no significant geographic differences among populations. These results supported the null hypothesis that E. japonicus within the Yellow Sea and East China Sea constituted a panmictic mtDNA gene pool. Based on the present study, the existence of separate genetic stocks in the Yellow Sea and East China Sea were not detected. Strong dispersal capacity of planktonic larvae, and ocean current could be the reasons for genetic homogeneity in this species in the Yellow Sea and East China Sea.     

Genetic diversity of mitochondrial D-LOOP sequences in the spotted scat (Scatophagus argus) from different geographical populations along the northern coast of the South China Sea

The genetic diversity of 289 spotted scat (Scatophagus argus) from seven populations along the northern coast of the South China Sea was studied by analyzing the full-length sequences of the mitochondrial control region (D-LOOP). The S. argus D-LOOP sequence was 1,004–1,010 bp long and contained 156 variant sites. The seven studied S. argus populations had a high degree of genetic diversity (haplotype diversity [Hd] = 0.99135; nucleotide diversity (π) = 0.01313). There was no obvious genetic differentiation among the seven geographical populations and gene exchange was frequent (Fst = −0.01867–0.01117, p > .05). Four distinct mitochondrial lineages were identified in the phylogenetic tree and the haplotype network. The between-lineage Fst was 0.71690–0.84940 (p < .001), but these lineages showed no obvious phylogeographic pattern. Based on D-LOOP mutation rates, we estimated that the four lineages diverged approximately 513,800–93,600 years ago, during the Eocene ice age, at which time falling sea levels may have led to population segregation. We estimated that S. argus population expansion occurred approximately 2.29–0.68 million years ago, during the late Pleistocene. During this period, sea levels rose again, allowing previously separated lineages to come into sympatry, which eventually gave rise to a highly genetically diverse population without pyhlogeographic structure. Here, we characterized the genetic structure and differentiation of seven S. argus populations from the northern coast of the South China Sea. Our results suggested that the seven S. argus populations from the northern coast of the South China Sea have a relatively low level of genetic variation and can be considered a single unit for the purposes of fishery development, utilization, and management.     

圆斑星鲽及相关种类线粒体DNA控制区结构分析

采用PCR产物直接测序法测定了圆斑星鲽(Verasper variegatus)的24个个体的线粒体控制区(Control region)核苷酸全序列, 并进行了结构分析。结果表明, 圆斑星鲽线粒体控制区核苷酸全序列具有长度多态性, 得到4种长度单元型, 主要表现为控制区中的串联重复序列的长度不同。对鲽形目鱼类如鲽科的条斑星鲽(Verasper moseri)、黄盖鲽(Limanda feruginea)、马舌鲽 (Reinhardtius hippoglossoides), 美洲拟庸鲽(Heppoglossoides platessoides )和鲆科的牙鲆(Paralichthys olivaceus)以及鳎科的欧洲鳎(Solea solea)、塞内加尔鳎(S. senegalensis)和沙鳎(S. lascari)的控制区的比较研究发现, 鲽形目鱼类的线粒体控制区均存在相似的结构, 即线粒体控制区可分为终止相关序列区(ETAS)、中央保守区(包括CSB-A、CSB-B、CSB-C、CSB-D、CSB-E、CSB-F)以及保守序列区(CSB1、CSB2、CSB3)和重复序列区(Repeat region)4个区域。通过与脊椎动物各个纲线粒体控制区序列的比较分析, 发现只有鲽形目(包括鲆、鲽类和鳎类)鱼类和两栖纲的无尾类在CSB-3之后存在相似的串联重复序列。     

Mitochondrial control region variability of baiji and the Yangtze finless porpoises,two sympatric small cetaceans in the Yangtze river

BaijiLipotes vexillifer (Miller, 1918) and the Yangtze finless porpoiseNeophocaena phocaenoides asiaeorientalis (Pilleri and Gihr, 1972) are two sympatric small cetaceans inhabiting the middle and lower reaches of the Yangtze River. In this study, a fragment (420–428 bp) of the mitochondrial control region was sequenced to provide the first comparative survey of genetic variability and population structure in these two endangered species, with samples of finless porpoises from the Yellow/Bohai Sea, East China Sea, and South China Sea also included. Low values of haplotype diversity and nucleotide diversity were found for both species, especially for the baiji and the Yangtze River and South China Sea populations of finless porpoises. The analysis of molecular variance (AMOVA) supported a high level of overall genetic structure among three porpoise populations in Chinese waters, with greatest differences found between either the Yangtze River population or the Yellow Sea population and the South China Sea population. The differentiation between the Yangtze and Yellow Sea populations was not significant, and the males have higher genetic differentiation than the females, suggesting a significant female-biased dispersal between these two populations. This study showed that the Yangtze finless porpoise, unlike the sympatric baiji, was not a genetically isolated population. The Yangtze and Yellow Sea porpoises should be included in the same management unit, but further studies using more samples and especially based on more molecular markers are urgently needed to confirm this.     

光裸方格星虫野生与养殖群体线粒体控制区序列的遗传差异分析

基于线粒体控制区序列对光裸方格星虫（Sipunculus nudus Linnaeus,1766）的2个养殖群体（营盘YP、竹林ZL）和4个野生群体（防城港FC、钦州QZ、大冠沙DG和越南海防YN）的91个个体进行遗传差异分析,研究光裸方格星虫养殖和野生群体的遗传变异情况。结果显示:获得的514 bp DNA序列中,野生与养殖群体的多态性位点数分别为82和60,均显示出对AT的偏倚性。共定义85个单倍型,共享单倍型4个,其中共享单倍型Hap5为原始单倍型,营盘群体均为独享单倍型。各群体的单倍型多样性（H_d）相同,野生群体的平均核苷酸多样性（P_i）（0.01531）略高于养殖群体（0.01514）,6个群体的遗传多样性水平依次为YN > YP > QZ > FC > ZL > DG。各群体间的遗传分化并不显著（P>0.05）,光裸方格星虫的遗传变异主要来自群体内个体间（99.08%）,同时未发现明显的地理谱系结构。研究表明,光裸方格星虫野生群体的遗传多样性水平总体略高于养殖群体;滩涂底播养殖方式较池塘养殖更利于维持光裸方格星虫遗传多样性;各群体间不存在显著的遗传分化,养殖群体正逐渐积累遗传变异,但尚未足够以形成其独立的遗传结构。     

基于线粒体Cyt b基因的龙头鱼群体遗传结构分析

在我国黄海、东海和南海海岸带上, 选取青岛、南通、舟山、三门、宁德、泉州和湛江共7个群体164尾龙头鱼(Harpadon nehereus)为研究对象, 通过PCR扩增共获得长度为1112 bp的线粒体DNA细胞色素b (Cyt b)基因序列, 共检测到32个变异位点, 其中单变异位点27个, 简约信息位点1个。164条序列定义了29个单倍型, 平均单倍型多样性(H_d)和核苷酸多样性(π)分别为0.3026±0.0479和0.000371±0.000379, 其中泉州群体的单倍型多样性和核苷酸多样性均最低。分析比较了不同龙头鱼群体间遗传变异情况, 发现群体间平均遗传距离为0.00035, 遗传分化指数F_ST值均小于0.05, 群体间没有发生明显的遗传分化。AMOVA分析结果显示龙头鱼群体遗传差异主要来源于群体内个体间的变异。中性检验的Tajima’s D和Fu’s F_s统计值均为负值且显著偏离中性, 核苷酸不配对分布图呈现明显的单峰分布, 表明龙头鱼历史上经历了群体扩张事件, 参考已知海洋鱼类Cyt b基因2%每百万年的进化速率, 估算群体扩张发生的时间大约在0.08—0.32百万年前的第四纪更新世中晚期。     

The impact of Yangtze River discharge on the genetic structure of a population of the rock bream,Oplegnathus fasciatus

As a rocky reef species, Oplegnathus fasciatus displays significant geographic differences that may be congruent with patterns of molecular variation. We used 10 polymorphism microsatellite loci to characterize the genetic structure and test the impact of ocean currents and the Yangtze River on the population genetic connectivity of the rock bream, O. fasciatus, sampled from Jiaonan (JN), Zhoushan (ZS) and Taiwan (TW). The average number of alleles was 17.78, with a total of 160 alleles, and the average allele value in the East China Sea (ZS?=?13.70) was higher than that in the Yellow Sea (JN?=?6.70). The individuals at the ZS location showed the highest expected heterozygosity (He?=?0.86), Nei's genetic diversity (H?=?0.84) and Shannon genetic diversity (I?=?2.21). Genetic bottleneck tests showed that individuals had experienced a recent bottleneck effect in the JN location. AMOVA analysis showed that 12.19% of genetic variation existed among populations, indicating significant population genetic differentiation in the species. The values of the population pairwise differentiation index (F_st and Φ_pt) ranged from 0.01 to 0.29. The UPGMA tree showed two diverged genealogical branches corresponding to sampling localities (Clade A?=?JN, Clade B?=?ZS and TW) in the species. The AMOVA and STRUCTURE analyses suggested that O. fasciatus may be composed of two stocks in the examined range, and the freshwater discharge from the Yangtze River might act as a physical barrier to genetic exchange between north and south China stocks of the species.     

Genetic diversity and population connectivity of the Asian green mussel Perna viridis in South China Sea,inferred from mitochondria DNA markers

The Asian green mussel Perna viridis is ecologically and economically important in the coastal regions of China. In order to characterize the genetic diversity and population connectivity of P. viridis in South China Sea, a 664 bp region of mitochondrial COI gene and a 293 bp region of 16S rRNA gene were sequenced and analyzed for 78 and 92 individuals from four populations in South China Sea, respectively. A total of 15 haplotypes were defined by 14 variable nucleotide sites in COI gene, and 7 haplotypes by 6 variable nucleotide sites in 16S rRNA gene. High haplotype diversity and low nucleotide diversity were observed in COI gene, while moderate haplotype diversity and low nucleotide diversity were observed in 16S rRNA gene. Pairwise F_ST values of COI gene were all negative and those of 16S rRNA gene ranged from −0.01409 to 0.10289. The results showed that no significant genetic divergence (or shallow genetic structure) and high levels of population connectivity among the four populations of P. viridis in South China Sea.     

Mitochondrial DNA differentiation between two forms of trout Salmo letnica, endemic to the Balkan Lake Ohrid, reflects their reproductive isolation

Mitochondrial haplotype diversity in sympatric populations of Ohrid trout, Salmo letnica was investigated by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mtDNA control region and ND1, ND3/4, ND5/6 segments. A 310 bp fragment at the 5' end, and a 340-572 bp fragment at the 3' end of the control region were sequenced from representatives of the populations studied. Based on pairwise comparison of the sequences, five new haplotypes were identified plus one identical with the brown trout Andalusian haplotype from the southern Iberian Peninsula. The combination of both RFLP and sequence data sets yielded a total of 10 composite haplotypes. A high degree of genetic subdivision between S. letnica typicus and S. letnica aestivalis populations was observed. The notion of a sympatric origin for the two morphs is discussed. Length variation of the mtDNA control region due to the presence of an 82 bp unit, tandemly repeated one to four times, in the region between the conserved sequence block-3 (CSB-3) and the gene for phenylalanine tRNA is reported. Further, we demonstrate that a single duplication of the approximately 82 bp repeat unit is a common element of the salmonid mitochondrial control region. The unique genetic structure of Ohrid trout represents a highly valuable genetic resource that deserves appropriate management and conservation.     

Genetic differentiation of Mediterranean horse mackerel (Trachurus mediterraneus) populations as revealed by mtDNA PCR-RFLP analysis

The genetic population structure of Mediterranean horse mackerel, Trachurus mediterraneus , from seven locations throughout the Black, Marmara, Aegean and eastern Mediterranean seas was investigated using restriction fragment length polymorphism (RFLP) analysis of the mtDNA 16S rDNA region. An approximately 2000-bp segment was screened in 280 individuals using six restriction enzymes, resulting in 10 composite haplotypes. The most common haplotype was present in 56.42% individuals; the next most frequent haplotype was present in 22.85% individuals. Average haplotype diversity within samples was moderate (0.38), and nucleotide diversity was low (0.00435). Mean nucleotide divergence for the seven sampling sites was 0.0028. Nucleotide divergence among samples was moderate, with the highest value detected between the Aegean Sea (Izmir) and the eastern Black Sea (Trabzon) populations (0.007055), and the lowest (−0.000043) between the Marmara Sea (Adalar) and the western Black Sea (Sile) populations. In Monte Carlo pairwise comparisons of haplotype frequencies, the Sinop from the middle Black Sea, Trabzon from the eastern Black Sea, and Iskenderun Bay from the north-eastern Mediterranean Sea exhibited highly significant (P   <   0.001) geographical differentiation from each other and from all other populations. Mantel's test indicated that the nucleotide divergence among populations of T. mediterraneus was not significantly associated with their geographical isolation ( r  = −0.2963; P   >   0.05). Consequently, the mtDNA 16S rDNA region provided evidence for the existence of three distinct T. mediterraneus populations (Sinop, Trabzon and Iskenderun Bay) in the Black and north-eastern Mediterranean seas.     

Differential population structuring of two closely related fish species, the mackerel (Scomber scombrus) and the chub mackerel (Scomber japonicus), in the Mediterranean Sea

Population genetic structures of the mackerel (Scomber scombrus) and chub mackerel (Scomber japonicus) were studied in the Mediterranean Sea. Fragments of 272 bp (S. scomber) and 387 bp (S. japonicus) of the 5'-end of the mitochondrial control region were sequenced from spawning individuals collected off the coasts of Greece, Italy, Spain, and Portugal. High levels of mitochondrial control region haplotypic diversity (> 0.98) were found for both Scomber species. Nucleotide diversity was higher in the mackerel (0.022) than in the chub mackerel (0.017). Global F(ST) values were also higher and significant in the mackerel (0.024, P < 0.0001) as opposed to the chub mackerel (0.003, P > 0.05). Molecular variance analyses showed differential genetic structuring for these two closely related species. There is extensive gene flow between Mediterranean Sea and Atlantic Ocean populations of chub mackerel, which are organized into a larger panmictic unit. In contrast, Mediterranean Sea populations of mackerel show some degree of genetic differentiation and are structured along an east-west axis. The analysed eastern Mediterranean Sea mackerel populations (Greece, Italy) are clearly separated from that of the western Mediterranean Sea (Barcelona), which forms a panmictic unit with eastern Atlantic Ocean populations. The genetic structures of both species showed asymmetric migration patterns and indicated population expansion.     

Genetic diversity of wild reindeer <Emphasis Type="Italic">Rangifer tarandus</Emphasis> L. from the eastern part of the Kola Peninsula: Polymorphism of the mtDNA control region

Based on the results of the analysis of nucleotide sequence polymorphism in the hypervariable fragment (left domain) of the mtDNA control region (D loop), the genetic diversity of wild reindeer from the eastern part of the Kola Peninsula (Murmansk oblast, Terskii region) was studied. Low values of the genetic diversity indices for Eurasian wild reindeer were detected. The effect of domestic reindeer breeding on the development of genetic diversity of the wild reindeer population in the studied region (indicating a low degree of hybridization of domesticated and wild reindeer) was estimated.     

Mitochondrial genetic diversity and population structure of a vulnerable freshwater fish,rock carp (Procypris rabaudi) in upper Yangtze River drainage

Rock carp (Procypris rabaudi (Tchang)) is an endemic species in the upper Yangtze River drainage. The genetic variability and population structure of eight populations (Mudong (MD), Hejiang (HJ), Wanzhou (WZ), Xishui (XS), Nantan (NT), Cangxi (CX), Tongjiang (TJ) and Wulong (WL)) were investigated based on mitochondrial DNA control region. According to the river dimensions, MD, HJ, WZ, WL and CX were defined as large river populations, while TJ, XS and NT were defined as small. The total genetic diversity level of rock carp was moderate (h = 0.858, π = 0.0079). Genetic differentiations among populations in small rivers were greater than those among populations in large rivers, and small rivers showed significant differentiation from large rivers. Analysis from the coalescent-based method showed that there were asymmetric gene flows among four large river populations (CX, HJ, MD and WZ). Demographic analyses suggested that the species as a whole and the WZ population experienced population expansion, while the HJ, NT, and MD populations experienced bottlenecks. The analysis of genetic diversity and population structure suggested that the WL, TJ, CX, NT and XS populations should be protected and managed separately, and that the HJ, MD and WZ populations should be considered as a genetic management unit.     

POPULATION GENETIC STRUCTURE OF FINLESS PORPOISES, NEOPHOCAENA PHOCAENOIDES, IN CHINESE WATERS, INFERRED FROM MITOCHONDRIAL CONTROL REGION SEQUENCES

Seven hundred and twenty base pairs (bp) of the mitochondrial control region from 73 finless porpoises, Neophocaena phocaenoides , in Chinese waters were sequenced. Thirteen variable sites were determined and 17 haplotypes were defined. Of these, 5 and 7 were found only in the Yellow Sea population and the South China Sea population, respectively, whereas no specific haplo-type was found in the Yangtze River population. Phylogenetic analyses using NJ and ML algorithm did not divide the haplotypes into monophyletic clades representing recognized geographic populations of finless porpoises in Chinese waters, suggesting the existence of migration and gene flow among populations. Analysis of molecular variance showed the obvious population genetic structure (φ_st= 0.41, P < 0.05); however, the structure was mainly between either the Yangtze River population or the Yellow Sea population and the South China Sea population. The genetic diversity (nucleotide diversity and haplotypic diversity) of the Yellow Sea population was significantly higher than those of the Yangtze River population and the South China Sea population, suggesting the relatively later divergence of the latter two populations and supporting the Yellow Sea population as the original center of Neophocaena .