Genetic and morphological variation of three freshwater lake populations of Coilia ectenes (Engraulidae)

Population structure of the important commercial fish, Coilia ectenes was investigated in samples from three freshwater lakes in the Eastern China using a multivariate approach of morphometries and mitochondrial DNA control region sequencing. A total of eighteen morphological distances of truss method and eight morphometric variables were taken from each fish. Multivariate analyses of the morphometric data revealed significant morphological differences among the three lake populations, especially for those samples from Taihu Lake. Discriminant functions were used to compare sites, and these permitted an 83% success rate in distinguishing fish from the three sites. However, no obviously geographical differentiation was found among those populations of C. ectenes based on the genetic data. In the AMOVA analysis, only 2.2% genetic variability came from different populations, and most of them were present within the sub-populations. Experience a recent population expansion and some movement of fish among those areas, quite possibly enough to bring about relative genetic homogeneity, but there is insufficient to prevent the three populations from differing phenotypically. The diversified environmental factors may be playing an important role in shaping morphological variations among those populations.     

A preliminary genetic distinctness of four Coilia fishes (Clupeiformes: Engraulidae) inferred from mitochondrial DNA sequences

In order to understand the genetic distinctness between four Coilia fishes, i.e. (C. ectenes, C. e. taihuensis, C. mystus, and C. grayii) the combined cytochrome b (cytb), 12S rRNA and 16S rRNA gene sequences were obtained and analysed. The results are as follows: (1) Neighbour-joining (NJ) and maximum parsimony (MP) trees were built with 1000 bootstrap replicates. They supported that Coilia fishes consisted of two clades: C. grayii form a monophyletic group, and the remaining samples form the other monophyletic group; (2) The topological differences between the NJ tree and the MP tree are checked by Templeton test, indicating that their differences are insignificant (P > 0.05); and (3) Kimura two-parameter distances between the samples of C. ectenes, C. e. taihuensis, and C. mystus all do not exceed 1.6%, and most of them are between 0 and 0.9%. Coilia ectenes, C. e. taihuensis, and C. mystus are closely related and their differences may be under subspecies. The text was submitted by the authors in English.     

A preliminary genetic distinctness of four Coilia fishes (Clupeiformes: Engraulidae) inferred from mitochondrial DNA sequences

In order to understand the genetic distinctness between four Coilia fishes,i.e., C. ectenes, C. e. taihuensis, C. mystus, and C. grayii, the combined cytochrome b (cytb), 12S rRNA and 16S rRNA gene sequences were obtained and analysed. The results are as follows: 1) neighbour-joining (NJ) and maximum parsimony (MP) trees were built with 1000 bootstrap replicates. They supported that Coilia fishes consisted of two clades: C. grayii form a monophyletic group, and the rest samples form the other monophyletic group; 2) the topological differences between the NJ tree and the MP tree are checked by Templeton test, indicating that their differences are insignificant (P > 0.05); and 3) Kimura-2-parameter distances between the samples of C. ectenes, C. e. taihuensis and C. mystus are all no bigger than 1.6%, and most of them are between 0.0% and 0.9%. C. ectenes, C. e. taihuensis and C. mystus are closely related and that their differences may be under subspecies.     

黄河裸裂尻鱼五种群mtDNA控制区的遗传结构

黄河裸裂尻鱼（Schizopygopsis pylzovi）主要分布于青藏高原东北部兰州以上黄河干支流水系和高原北部柴达木内流水系,由于其对高原特殊生境表现出独特的适应性而受研究者关注。本文采用聚合酶链式反应（PCR）和直接测序的方法获得了一个分布于柴达木内流水系种群和四个黄河支流种群共99条个体的线粒体DNA（mtDNA）控制区（D-100p）821bp核苷酸的序列,探讨了其种群遗传结构和分化。99条个体的序列经比对后,发现77个（9．37％）多态性位点,共定义了53个单倍型,其中有一个单倍型（DT12）为两个种群（大通种群和互助种群）所共享,其余52个单倍型均为某个种群所特有。遗传多样性分析表明,柴达木种群的单倍型多样度（h=0．79±0．06）和核苷酸多样度（π=0．0027±0．0017）均略低于黄河支流各种群。AMOVA分析结果显示,遗传差异主要发生在种群之内,占75．39％,而小同水系的种群间只有24．61％,各种群间成对的Fst及遗传距离均表明种群间出现了一定程度的种群分化,但系统进化树最示出一种混杂的单倍型分布格局,提示青藏高原东北部地质事件所造成的水系间的隔离形成较晚。单倍型歧点分布呈现为单峰以及中性检验Tajima的D（-1．497,P=0．058）和Fu的Fs（-24．741,P=0．001）结果综合表明,黄河裸裂尻鱼在青藏高原隆升过程中曾经历过近期的种群扩张事件[动物学报54（6）：972—980,2008]。     

Genetic diversity of Setipinna taty (Engraulidae) populations from the China Sea based on mitochondrial DNA control region sequences

The genetic diversity of Setipinna taty, which is commercially fished in the China Sea, was studied based on mitochondrial DNA control region sequences. PCR was used to amplify the control region fragment in 100 individuals of S. taty collected from Weihai (WH), Yantai (YT), Zhoushan (ZS), Xiangshan (XS), and Ninghai (NH) in China. A control region fragment of 656 bp was successfully sequenced in these 100 individuals. The A+T content of this S. taty control region was 71.7%; 172 variable sites and 62 haplotypes were found. Nucleotide diversity in the WH, YT, ZS, XS, and NH groups was 0.0228, 0.0247, 0.0441, 0.0126, and 0.0238, respectively. The haplotype diversity was 0.984, 0.911, 0.989, 0.926, and 0.979, respectively. Analysis of molecular variance showed that 97.95% of genetic variation was within populations, and only 2.05% among populations. The neighbor-joining phylogenetic tree obtained based on genetic distance showed that no significant genealogical structure exists throughout this range of S. taty. These results indicate no apparent geographical differentiation in the comparison of Yellow Sea and East China Sea populations of S. taty. Within the control region, we identified an extended termination-associated sequence domain, a central conserved sequence block domain and a conserved sequence block domain; insertions of short tandem repeat sequence segments were found at the 5' end of the control region.     

基于线粒体控制区的序列变异分析中国东南部沿海凤鲚种群遗传结构

为了解中国东南部沿海凤鲚(Coilia mystus)的种群遗传多样性和遗传结构, 本文分析了长江口(CJ)、钱塘江口(QT)、闽江口(MJ)和九龙江口(JL)4个凤鲚地理群体的mtDNA控制区561 bp片段的序列变异。65尾样本共检测到28个单元型。4个群体总的单元型多样性和核苷酸多样性均较高(h = 0.9433 ± 0.0168, π = 0.0317 ± 0.0158), 但单个群体的核苷酸多样性水平却很低, 其中以CJ最高(π = 0.0080 ± 0.0046), MJ最低(π = 0.0015 ± 0.0013)。MJ与JL群体之间以及CJ与QT群体之间的平均K2P遗传距离很小, 分别为0.3%和0.8%; 而CJ、QT分别与MJ、JL群体之间的遗传距离均较大, 达到了6%。采用最大似然法(ML)、最大简约法(MP)和邻接法(NJ)分别构建的单元型间的系统发育树揭示, 4个凤鲚群体构成CJ-QT 和MJ-JL 2个支系, 且具有极高的支持率。单元型的网络分析也显示这两个支系间有高达28步的突变次数。AMOVA分析显示大部分的遗传变异来自这两支系群体间(90.77%), 表明凤鲚群体间存在着显著的地理分化。种群分化指数和基因流分析也表明, 支系间群体有着明显的遗传分化(F_ST > 0.9, N_m < 0.03)。所有分析结果支持所研究的凤鲚标本属于两个不同的地理种群, 且种群的分化至少已达到亚种水平。采用BEAST和TRACER软件得到凤鲚两个亚种的最近共同祖先约在0.34–0.46百万年前, 处于更新世晚期, 推测可能是第四纪晚期的气候旋回和海平面的升降导致了凤鲚的种群分化。     

Mitochondrial DNA diversity of Coilia mystus (Clupeiformes: Engraulidae) in three Chinese estuaries

We used sequences of mitochondrial cytb and 16SrRNA gene segments in order to clarify the genetic diversity and population structure in three Chinese estuary populations of Coilia mystus: 21 individuals from ChangJiang River (Yangtze River) estuary, 22 from MinJiang River estuary, and 22 from ZhuJiang River (Pearl River) estuary (65 individuals total). We obtained 607 base pairs of consensus cytb sequence. Thirty four distinct haplotypes were detected among the 65 cytb sequences. The indexes of nucleotide diversity (π) in these three populations were ChangJiang 0.533%, MinJiang 1.135%, and ZhuJiang 0.268%. MinJiang is the largest of the three populations. Genetic distances within the populations were between 0.3 and 1.2%, and 0.8 to 10.8% among populations. The largest genetic distance was 10.8% between the ChangJiang and ZhuJiang populations, and the smallest was 0.8% between MinJiang and ZhuJiang populations. Analysis of molecular variance (AMOVA) analysis revealed that variation among populations accounts for 90.25% of total variation, suggesting that this is the main source of total variance. We obtained 470 base pairs of consensus sequence of 16SrRNA. We detected 19 distinct haplotypes among the 65 sequences. The indexes of nucleotide diversity (π) in these three populations were ChangJiang 0.108%, MinJiang 0.843%, and ZhuJiang 0.097%. MinJiang is also the largest among these three populations. Genetic distances were between 0.1 and 0.9% within populations and 0.5 to 1.9% between populations. The largest genetic distance was the 1.9% between the ChangJiang and MinJiang populations, and the smallest was 0.5% between the MinJiang and the ZhuJiang populations. AMOVA analysis disclosed that variation among populations accounts for 74.61% of total variation, suggesting that this is the main source of total variation. The results of this study suggest that the three Coilia mystus populations, especially the most isolated Changjiang population, have developed significant genetic structure.     

喜马拉雅旱獭线粒体DNA控制区遗传多样性及系统发育

喜马拉雅旱獭是青藏高原的优势种,数量多、分布广,全面了解其遗传背景对该地区旱獭资源的保护与合理利用具有重要的意义。本研究以青藏高原云南、西藏和青海三省区共13个地理种群计258只旱獭为研究对象,PCR扩增获得线粒体DNA控制区基因部分序列(887 bp),并运用种群遗传学方法进行遗传多样性分析。结果显示:258份样品共发现了84个变异位点(9.40%),定义了68种单倍型,其单倍型多样性(h)平均值为0.968±0.003、核苷酸多样性(π)平均值为0.017 25±0.016 37,种群总体遗传多样性较高。AMOVA方差分析显示13个地理种群间存在着明显的遗传分化(Fst=0.620 67,P<0.001),种群间基因交流多数较低(Nm<1)。基于单倍型构建的系统发育树中13个地理种群的喜马拉雅旱獭聚为两支,其中来自青藏高原西南地区(西藏安多、青海格尔木、青海囊谦、云南迪庆)的18个单倍型聚成一个大的分支(A支),其余50个单倍型聚为一个大的分支(B支),在NETWORK网络图中也可见到相似网络拓扑结构。研究结果显示青藏高原喜马拉雅旱獭种群以唐古拉山脉为界分为两个大的种群,说明地理隔离是影响喜马拉雅旱獭种群动态变化的主要因素。     

Isolation and characterization of microsatellite loci in the fish Coilia mystus (Clupeiformes: Engraulidae) using PCR-based isolation of microsatellite arrays

Coilia mystus is the most important harvested fish species in China; it inhabits quite different water environments during the different life history stages. Populations of C. mystus have dropped sharply due to overharvesting and water pollution. We developed eight microsatellite loci in C. mystus for conservation genetics studies. These new markers were tested in 20 individuals from the Min River in ChangLe. The number of alleles ranged from 3 to 8, the expected heterozygosity from 0.621 to 0.853 and the observed heterozygosity from 0.473-0.800. Only two loci deviated significantly from Hardy-Weinberg expectations due to heterozygote deficiency. These primers may provide a tool for understanding demography and population structure of this economically important and threatened species.     

四川地区猕猴线粒体DNA 控制区遗传多样性及其种群遗传结构

猕猴是最理想的医学实验灵长类动物,并且是国家二级保护动物。四川地区的猕猴数量多、分布广,全面了解其遗传背景对于该地区猕猴资源的保护与合理利用具有重要的意义。本研究对来自四川8 个地理种群的231个不同猕猴个体的线粒体DNA 控制区部分序列进行了测定和群体分析,发现了110 个变异位点(22. 49% ),定义了56 种单倍型,其单倍型多样性(h)平均值为0. 686、核苷酸多样性(π)平均值为0. 01483,种群总体遗传多样性较高;进一步分析表明,8 个地理种群间存在着明显的遗传分化(F_st = 0. 70412,P < 0. 05),种群间基因交流较低(N_m < 1);系统发育树显示,四川猕猴8 个地理种群的单倍型基本上成簇分布在系统树上,与地理位置呈现一定的对应关系,说明四川猕猴具有明显的系统地理分布格局。地理隔离和人类活动可能是促使四川猕猴种群分化的主要因素。     

Genetic structure and differentiation of four populations of Afghan Pika (Ochotona rufescens) in Iran based on mitochondrial cytochrome b gene

The population genetics of the Afghan Pika (Ochotona rufescens) was studied in Northern Khorasan Province, Iran. For prediction of the genetic differentiation of four populations, the DNA of mitochondrial cytochrome b of 32 individuals from four areas was sequenced and a Bayesian analysis based on the HKY model was constructed. In total, 15 polymorphic sites, 1125 conserved sites (98.7%) and 14 different haplotypes were found. The phylogenetic tree resulting from the Bayesian analysis and network analysis showed that all samples were clustered in two major groups and the haplotypes of the four populations did not separate geographically. An analysis of molecular variance (AMOVA) indicated that a large majority of the genetic variance was due to the variance within populations. The results of fixation indices showed significant genetic structure among populations in both methods. The pairwise F_st revealed that two northern populations have a significant genetic differentiation from two southern populations, but no significance pairwise F_st value was demonstrated between the closed populations. Nei's genetic distances between closed populations were not significant, while significant values occurred between distant populations. It seems that there is not a major discontinuity between populations of Afghan Pika based on cyt-b mitochondrial gene. However, phylogenetic analysis did not separate populations and a large majority of the genetic variance was found within populations. However, AMOVA analysis showed a significant level of genetic structure among populations (p?<?0.001) and between groups (p?<?0.5). It seems that these results suggest shallow genetic differentiation between populations of different geographic groups.     

Genetic population structure of Hemigrammocypris rasborella (Cyprinidae) inferred from mtDNA sequences

The genetic population structure of the small cyprinid Hemigrammocypris rasborella, distributed widely in lowlands of western Japan, was examined using partial sequence data of mitochondrial DNA (mtDNA). Molecular phylogenetic analysis revealed that the populations of the western Kyushu region were markedly differentiated from all eastern populations, such that the groups would be comparable to different species; their divergence was inferred to have occurred in the Late Miocene–Pliocene. Also, a largely divergent mtDNA group (with divergence in the early Pleistocene) was found in the Sanyo and northeastern Shikoku regions, forming a secondary contact zone in the western Kinki with the eastern mtDNA group. To date, these aspects of the population structure of H. rasborella appear to be unique among lowland fishes in western Japan. Deeper understanding of the formation processes of freshwater faunas in western Japan will require further comparisons of the phylogeographic patterns and ecological traits of constituent species.     

Reconstructing habitat use of Coilia mystus and Coilia ectenes of the Yangtze River estuary, and of Coilia ectenes of Taihu Lake, based on otolith strontium and calcium

The habitat use and migratory patterns of Osbeck's grenadier anchovy Coilia mystus in the Yangtze estuary and the estuarine tapertail anchovy Coilia ectenes from the Yangtze estuary and Taihu Lake, China, were studied by examining the environmental signatures of strontium and calcium in their otoliths using electron probe microanalysis. The results indicated that Taihu C. ectenes utilizes only freshwater habitats, whereas the habitat use patterns of Yangtze C. ectenes and C. mystus were much more flexible, apparently varying among fresh, brackish and marine areas. The present study suggests that the spawning populations of Yangtze C. ectenes and C. mystus in the Yangtze estuary consist of individuals with different migration histories, and individuals of these two Yangtze Coilia species seem to use a variety of different habitats during the non-spawning seasons.     

Genetic differentiation and structure of Hippocrepis valentina (Leguminosae) populations

We present an analysis of isozyme variability in natural populations of the plant species Hippocrepis valentina (Leguminosae), which is endemic to the eastern Mediterranean coast of Spain and currently has endangered species status. Our results, obtained by starch-gel electrophoresis of 15 loci, show normal levels of variability for species with similar biology. The comparison with the patterns of genetic variability of two closely related species, H. balearica and H. grosii, confirms the taxonomic status of H. valentina as a proper species, independent of H. balearica, as previously suggested. The analysis of population subdivision shows that substantial variation among populations is present, and a hierarchical analysis demonstrates that when zones are defined according to their geographic location, a higher differentiation among populations within zones than among zones is found. Indirect estimates of gene flow indicate that levels of migration per generation are relatively low, except for a group composed of four populations, three of which are in close proximity. For the remaining populations, there is evidence of substantial differentiation. These results have implications for the design of a conservation strategy for this species.     

从线粒体控制区全序列变异看短颌鲚和湖鲚的物种有效性

经克隆测序获得我国七丝鲚(Coilia grayii)、凤鲚(C. mystus)、刀鲚(C. nasus)和短颌鲚(C. brachygnathus)以及太湖湖鲚(C. nasus taihuensis)等4个种和1亚种32尾个体的mtDNA D-loop区全序列, 以日本鳀(Engraulis japonicus)和秘鲁鳀(E. ringens)为外类群构建了中国鲚属的分子系统发育树, 并讨论了短颌鲚和湖鲚的物种有效性。结果显示, 七丝鲚的D-loop区全序列长1,208 bp, 凤鲚1,279–1,361 bp, 刀鲚1,252–1,290 bp, 短颌鲚1,214–1,252 bp, 湖鲚1,252–1,442 bp, 除七丝鲚外的其他种类个体间均表现出序列长度的多态性。短颌鲚、刀鲚和湖鲚三者间的平均K 2-P遗传距离仅为0.011–0.020, 明显小于它们与凤鲚、七丝鲚及外类群间的遗传距离(0.051–0.349)。以邻接法和最大简约法构建的系统发育树表明, 刀鲚、短颌鲚及湖鲚均未各自构成单系, 而是共同构成一个单系群, 三者并未发生显著分化。研究表明, 短颌鲚和湖鲚为刀鲚的淡水生态型种群, 并非有效物种。系统发育分析表明, 中国鲚属3个有效物种间以凤鲚最为原始, 刀鲚和七丝鲚为姐妹群, 处于较进化的位置。推测凤鲚可能是鲚属祖先种最早从起源中心扩散到西北太平洋的后裔, 而刀鲚和七丝鲚则是凤鲚在演化过程中分别适应寒冷和温暖气候而分化出的物种。     

Genetic diversity and population structure analysis of Qinghai-Tibetan plateau schizothoracine fish (Gymnocypris dobula) based on mtDNA D-loop sequences

Gymnocypris dobula is a commercially important fishery species and mostly distributed in the freshwater of Tibetan plateau. In this study, genetic diversity and intraspecific population differentiation were examined by using mitochondrial DNA D-loop sequences in 97 individuals sampled from three localities (Pali, Lasa and Yanghu) in the Tibetan plateau. Two hundred and fourteen polymorphic sites and 50 haplotypes were defined among the three localities. Genetic diversity analysis showed that the highest genetic diversity level was found in Pali population. Phylogenetic relationships analysis results indicated that closer phylogenetic relationships were found between the Yanghu and Lasa populations. Genetic population differentiation analysis indicated that the majority of variation (84.91%) was attributed to variations among populations and the largest differentiation was found between Pali and Lasa localities (F_st = 0.874, Nm = 0.036). Moreover, the historical demographic events were assessed by implementing the mismatch distribution analysis, Fajima's D test and Fu's Fs test. The results indicated that the Pali population had undergone a demographic expansion, possibly within the last 0.163 MYA (Million Years Ago). Our study firstly identified the population genetic structure of the G. dobula, which could be helpful for artificial breeding, fishery stock identification and resource conservation for this species.     

Phylogeographic population structure of great reed warblers: an analysis of mtDNA control region sequences

A portion of the mitochondrial control region (494 bp) was sequenced in 106 great reed warblers sampled in six breeding populations in Europe and one wintering population in Africa. In total, 33 different haplotypes were found. There was little evidence of divergence between populations in northern and western Europe whereas the sample from Greece differed significantly from the other European breeding populations. The lowest haplotypc diversity was found near the distribution range limit in Sweden and in The Netherlands suggesting recent effects of bottlenecks/founder events in these areas. A neighbour-joining analysis of the different haplotypes placed the haplotypes into two distinctive clades, A and B. The divergence of the two clades was on average 1.29%. Accounting for the within cladc variation suggested a divergence time between these lines approximately 70 000 years BP. The frequency of the two clades changed longitudinally across Europe with the A haplotypc in the west and the B haplotypc in the east. All birds from Kenya carried the B haplotypc suggesting an origin of these birds east of Latvia/Greece. The long-term female effective population size was estimated to be 20 000 individuals, which is approximately 2% of current population size.