单环刺螠(Urechis unicinctus)微卫星标记开发及5个地理种群遗传结构分析

采用高通量测序法从单环刺螠基因组中开发微卫星标记,并通过一个秦皇岛单环刺螠野生群体对其进行多态性评价,利用获得的多态性引物对秦皇岛、烟台、潍坊、青岛、大连5个不同海区的野生单环刺螠群体进行了遗传多样性及遗传结构分析。结果表明:本实验设计的50个微卫星标记能稳定扩增的有38个,其中22个微卫星位点在5个群体中均表现为高多态性,等位基因数(Na)介于24—44之间,多态信息含量(PIC)介于0.921—0.967之间。5个群体总的平均有效等位基因数(N_e)范围为6.629—8.850,平均观测杂合度(Ho)和平均期望杂合度(He)范围分别为0.790—0.912、0.851—0.896,大连群体的杂合度最大(H_e=0.8962),潍坊群体的最小(H_e=0.8510),其中有12个微卫星位点在不同群体中出现显著偏离Hardy-Weinberg平衡的现象。平均遗传分化指数(FST)表明,群体分化水平处于中等分化水平(FST值为0.0880—0.1136);聚类分析显示烟台群体先与青岛群体聚为一支,再与秦皇岛群体聚类,然后跟潍坊群体聚为一支,大连群体单独聚为一支;遗传距离模式(IBD)显示单环刺螠群体的地理距离与遗传距离间不存在显著的线性关系。本研究开发的22对微卫星标记可以用于单环刺螠遗传结构分析研究,为下一步单环刺螠种质资源保护和人工繁育提供参考依据。     

Potential transport of plaice eggs and larvae between two apparently self-contained populations in the Irish Sea

A coupled physics particle-tracking model, driven by realistic meteorological forcing was used to examine the dispersal and transport of plaice eggs and larvae in the year 2000 from two spawning grounds in the Irish Sea. The model included passive transport of eggs and early stage larvae, diel vertical movements for larvae between 7 and 10.5 mm in body-length and tidally synchronised, vertical movements for larger larvae (>9 mm body-length). The year 2000 was chosen because of the availability of ichthyoplankton data with which to initialise the model. The majority of larvae originating from spawning in the eastern Irish Sea settled into nursery grounds along the Scottish, English and Welsh coasts, in agreement with previous findings. In contrast, a significant portion of larvae originating from spawning in the western Irish Sea was transported eastwards to these same nursery grounds. Transport across the Irish Sea resulted from the onset of tidally synchronised vertical behaviour encoded in the model for older larvae. Settlement of larvae into local nursery grounds along the Irish coast was limited. Because of the prevailing winds and currents in the region, plaice eggs and larvae are unlikely to be transported from east to west; in most years spawning in the western Irish Sea probably acts as an additional source of juveniles for nursery grounds along the Scottish, English and Welsh coasts.     

许氏平鲉4 个野生群体遗传多样性微卫星分析

利用18对微卫星引物分析了荣成湾群体1、荣成湾群体2、即墨群体和龙口群体共4个许氏平鲉(Sebastes schlegeli)野生群体的遗传多样性。结果表明:4个群体的平均有效等位基因数分别为3.1、2.7、2.8和2.7,平均期望杂合度分别为0.590、0.540、0.552和0.491,平均多态信息含量分别为0.577、0.495、0.538和0.528。4个许氏平鲉群体均表现出较高的遗传多样性水平,但也都存在杂合子缺失现象。Χ2检验显示,4个群体中大部分微卫星位点偏离Hardy-Weinberg平衡(P<0.05)。群体间Fst值为0.04,说明遗传变异主要存在于群体内,群体间的遗传分化程度较低。该研究结果将为许氏平鲉种质资源保护和合理利用以及人工增殖提供理论参考。     

斜带石斑鱼不同地理群体遗传变异的微卫星分析

筛选5时可分析引物对斜带石斑鱼(Epinephelus coioides)台湾群体和广东群体的DNA多态性进行分析.根据电泳检测结果,平均每个引物获得4.2个等位基因,未发现群体的特异位点.获得2个群体平均观测杂合度(H_o)分别为0.672 1和0.682 6,平均期望杂合度(H_e)分别为0.647 9和0.544 4,两群体间的遗传分化系数(F_(ST))为0.011 0.统计结果表明,2个群体有着较高的遗传变异性,群体间的遗传分化程度处于多数海水鱼类的平均水平.     

Continental shelf nurseries and recruitment variability in American plaice and yellowtail flounder on the Grand Bank: insights into stock resiliency

In 1994, a directed fishing moratorium was declared for Grand Bank American plaice (Hippoglossoides platessoides) and yellowtail flounder (Limanda ferruginea) stocks because both stocks showed severe declines in abundance from heavy exploitation during the mid 1980s and early 1990s. Four years later, the fishery for yellowtail re-opened while the plaice stock has shown little recovery and the moratorium is still in effect. To assess the possible causes of the differences in recovery between species, we examined the spatial structure and environmental characteristics of the continental shelf nursery habitats of plaice and yellowtail, and their relationship to recruitment variability and overall population size. Depth plays a major influential role determining the spatial pattern and the abundance of juveniles of both species and in the case of plaice the spatial structure of the adult population also determines the amount of nursery area utilised by juveniles. Recruitment variability was higher in plaice than in yellowtail. We found year class synchrony in both species indicating that common environmental conditions and/or biological processes are affecting recruitment in a similar manner. Density-dependent regulation appears to be more severe in yellowtail and this should contribute to a more stable population when compared to plaice. These results are discussed in terms of resiliency of both stocks to over-exploitation.     

Shifts in the timing of spawning in sole linked to warming sea temperatures

Phenotypic traits such as peak spawning time may vary within and differ between populations in relation to environmental factors, such as temperature. Sole (Solea solea) is a valuable, commercially exploited species that spawns in late winter or spring. The date of peak spawning was estimated for each year for seven stocks from monthly fish samples collected from commercial fisheries since 1970. Four out of seven stocks showed a significant long-term trend towards earlier spawning (Irish Sea, east-central North Sea, southern North Sea, eastern English Channel) at a rate of 1.5 weeks per decade. The other three stocks (Bristol Channel, western English Channel and western-central North Sea) failed to show a relationship, but the available time series were limited for these stocks (< 10 years). Sea surface temperature during winter significantly affected the date of peak spawning, although the effect differed between stocks. The implications of the effect of winter temperature on the timing of spawning for the population dynamics are discussed.     

Population structure of the thornback ray (Raja clavata L.) in British waters

Prior to the 1950s, thornback ray (Raja clavata L.) was common and widely distributed in the seas of Northwest Europe. Since then, it has decreased in abundance and geographic range due to over-fishing. The sustainability of ray populations is of concern to fisheries management because their slow growth rate, late maturity and low fecundity make them susceptible to exploitation as victims of by-catch. We investigated the population genetic structure of thornback rays from 14 locations in the southern North Sea, English Channel and Irish Sea. Adults comprised < 4% of the total sampling despite heavy sampling effort over 47 hauls; thus our results apply mainly to sexually immature individuals. Using five microsatellite loci, weak but significant population differentiation was detected with a global F_ST = 0.013 (P < 0.001). Pairwise F_st was significant for 75 out of 171 comparisons. Although earlier tagging studies suggest restricted foraging distances from home areas, the absence of genetic differentiation between some distant populations suggests that a substantial fraction of individuals migrate over wide areas. Autumn/winter locations appear to have a lower level of differentiation than spring/summer, which could be due to seasonal migration. Management and conservation of thornback ray populations will be challenging as population structure appears to be dynamic in space and time.     

Isolation via enrichment and characterization of ten polymorphic microsatellite loci in the cuttlefish, Sepiella maindroni de Rochebruns

Ten novel polymorphic microsatellite loci were developed using an enrichment and colony hybridization protocol and characterized for the cuttlefish, Sepiella maindroni. Polymorphism was explored by using 30 individuals from the coastal waters of Zhanjiang, Guangdong province, China, in December 2007. The number of alleles ranged from 5 to 13. The ranges of observed and expected heterozygosity were from 0.578 9 to 1.000 0 and 0.682 8 to 0.925 7, respectively, and the average polymorphic information content (PIC) was 0.778 5. These microsatellite loci will certainly facilitate the detection of the genetic variation and population structure of S. maindroni.     

Genetic variability of the blue and red shrimp Aristeus antennatus in the Western Mediterranean Sea inferred by DNA microsatellite loci

Genetic variation at eight microsatellite loci was studied in nine populations of the blue and red shrimp Aristeus antennatus to investigate whether distinct stocks are present in the Western Mediterranean Sea. A high level of gene flow and no evidence of genetic partitioning were discovered. No significant variation was found (F_ST = 0.00673, P-value = 0.067) even when shrimps from exploited and those from deep-water unexploited grounds were compared. No evidence of reduction or expansion of population size in the recent past was found, as indicated by the bottleneck and interlocus g-tests. Our results are consistent with previous studies using mitochondrial gene methods and allozymes, indicating that, for this species, extensive pelagic larval dispersal and adult migration are probably responsible for the genetic homogeneity observed. In particular, due to a different bathymetric distribution of males and females, reported to be associated with different water masses and hence with possible differential dispersal capacity between sexes, the hypothesis of sex-biased dispersal was tested. Mean values of corrected assignment indices and mean relatedness values were higher for males, suggesting that females are the more widely dispersing sex. Molecular assessment of A. antennatus from the Western Mediterranean provides data of biological and evolutionary interest for the successful management of such a highly valuable fishery resource.     

基于SSR标记的魁蚶(Scapharca broughtonii)不同群体遗传结构的分析

运用6对微卫星分子标记分析了山东蓬莱(SP)、山东黄岛(SH)、江苏前三岛(JQ)、韩国统营(Korea)等魁蚶4个地理群体的遗传结构。结果表明,4个魁蚶群体在6个位点的等位基因数(a)为11.3333—14.3333,平均有效等位基因数(ae)为6.2977—9.5873;平均杂合度观察值(Ho)为0.4083—0.5641,平均杂合度期望值(He)0.7849—0.8753,各位点平均多态信息含量(PIC)值为0.7703—0.9102,遗传多样性非常丰富。聚类分析结果表明黄岛群体与前三岛群体聚在一起,蓬莱群体与统营群体聚在一起最后聚类。本研究结果可为今后魁蚶种质资源保护和遗传育种提供遗传学资料。     

尼罗罗非鱼Oreochromis niloticus、奥利亚罗非鱼O.aureus和红罗非鱼O.sp.群体遗传多样性的比较

用9对微卫星引物对尼罗罗非鱼Oreochromis niloticus、奥利亚罗非鱼O.aureus和红罗非鱼O.sp.群体的遗传变异进行了比较研究。在3个群体109个个体中共检测到60个等位基因,3个群体的平均等位基因数分别为4.11、1.33和3.44,平均观测杂合度分别为0.528、0.056和0.491,平均期望杂合度分别为0.644、0.091和0.526,平均多态信息含量分别为0.580、0.077和0.466。杂合子偏离度D值分别为0.148、0.222和0.044,表明3个罗非鱼群体存在不同程度的杂合子缺失。卡方检验表明3个群体的大部分位点偏离Hardy-Weinberg平衡,存在遗传漂变现象。群体间遗传分化显著(遗传分化指数FST在0.329到0.656之间,P<0.01)。尼罗罗非鱼和红罗非鱼群体间的遗传距离最小(0.47)。上述分析表明,尼罗罗非鱼的遗传多样性最高,奥利亚罗非鱼的遗传多样性最低,群体间分化显著。表明尼罗罗非鱼和红罗非鱼尚具有一定的选育潜力,而奥利亚罗非鱼遗传多样性低,不利于选择育种,需要引进新的种群。     

The food and growth of 0-group flatfish on nursery grounds in the Clyde Sea Area

In early summer plaice (Pleuronectes platessa L.) in the Clyde Sea Area eat Tellina siphons and spionid palps. As they grow, they take progressively larger siphons, fragments of spionids and some crustaceans. In late summer they eat entire spionids and other sedentary polychaetes. There is no significant predation on fish. The common dab [Limanda limanda (L.)] took a wider range of prey species than the plaice, but also depended mainly on polychaetes, crustaceans and bivalves. Depth distribution may limit both intra- and interspecific competition between these fish and also influence the choice of prey. The results from the main nursery areas of Irvine and Ayr Bays were similar to those from the other bays throughout the Clyde Sea Area and support earlier work at Firemore Bay in north-west Scotland and in the Dutch Wadden Sea.The growth rate of the fish increases during the early summer and may be related to increasing food availability, feeding experience of the fish and longer day length. Growth rate between areas appeared to be correlated with benthic productivity. 0-group plaice in Irvine, Ayr and Firemore Bays grew to different mean lengths by the end of the summer growth season, being 82 mm, 73 mm and 61 mm, respectively, for the period 1972–1974. This agrees with the known differences in standing stocks of intertidal macrobenthos in these bays (17·0, 11·1 and 1·3 g m^?2 dry weight, respectively). The significance of this in relation to survival is discussed. Offshore migration of the larger fish in the autumn may result in apparent slowing of growth rates prematurely in inshore areas. The year-class strength depends more on the food availability in the nursery areas than on the number of plaice that settle on the bottom.     

A comparative analysis of recruitment variability in North Atlantic flatfishes — testing the species range hypothesis

The hypothesis that recruitment variation in flatfishes should be most variable at the northern edge of the species range, least near the centre of the range, and intermediate near the southern limit was tested using stock and recruitment data generated from sequential population analysis for several different flatfish stocks involving four species (plaice Pleuronectes platessa, sole Solea vulgaris from the eastern Atlantic, American plaice Hippoglossoides platessoides, and yellowtail flounder Limanda ferruginae from the western Atlantic). Several groundfish species have been found to conform to this so-called species range hypothesis with the suggestion that density-independent processes predominate at the edges of the distributional range and density-dependent processes dominate in the centre of the range. Our results were generally inconsistent with the hypothesis: the coefficient of variation (CV) of recruitment for plaice in the eastern Atlantic was independent of latitude, the CV of recruitment for sole exhibited a dome-shaped relationship with latitude with the highest CVs occurring at the mid-point of the range, and the CV of recruitment for the western Atlantic stocks exhibited a monotonic decrease with latitude. We extended our latitudinal analyses by assessing both the degree of dependency of recruitment on spawning stock biomass and the spatial and temporal scales of variability in recruitment and pre-recruit survival for the eastern Atlantic stocks. In general our analysis revealed no evidence of a strong stock and recruitment relationship for any of the stocks examined, and previously published analyses revealed no such patterns with latitude. Analysis of both de-trended recruitment and pre-recruit survival time series over the species ranges of sole and plaice revealed strong positive correlations among adjacent stocks and inverse correlations among stocks at the extremes of the range. Recruitment variation in the flatfish stocks examined appears to be dominated by density-independent factors, operating at a local scale, on the egg and larval stages.     

多鳞(Sillago sihama)鱚4个野生地理群体的微卫星标记分析

采用微卫星DNA分子标记方法对中国南部沿海多鳞鱚(Sillago sihama)广西北海、广东湛江、汕尾及海南三亚4个野生地理群体进行了遗传多样性及遗传分化等研究。结果表明:(1)用于研究的34对微卫星引物在4个群体中具有丰富的多态性,有效等位基因数介于1.71—20.46之间,多态信息含量PIC介于0.40—0.95之间。4个群体总的平均期望杂合度(He)为0.598,观测杂合度(Ho)为0.572,SY群体的杂合度最大(He=0.606,Ho=0.623),SW群体的最小(He=0.586,Ho=0.508)。(2)群体间F-统计量分析表明,群体遗传分化处于高度分化水平(Fst平均值为0.306);基于奈氏遗传距离(DA)构建的NJ和UPGMA聚类树均显示地理位置相邻的群体聚在一起。     

石鲽野生群体和养殖群体遗传多样性的ISSR分析

采用简单序列重复区间(ISSR)技术,对石鲽的3个野生群体——青岛群体(QD)、威海群体(WH)、莱州群体(LZ)和一个养殖群体(YZ)进行了遗传多样性研究.结果表明,从60个ISSR引物中筛选出8个ISSR引物对四个群体的基因组DNA进行扩增,共获得183个重复性好且5谱清晰的位点,4个群体的多态位点比例为48.1%~50.8%,Shannon多样性值为15.68~18.53,群体内个体间遗传相似度为0.9053~0.9189,群体间的遗传距离为0.0055~0.0173.同其他鱼类相比,石鲽群体的遗传多样性水平较低.用UPGMA方法构建的群体进化树显示,WH群体和LZ群体遗传距离最近,首先聚在一起,两者然后与YZ群体聚类,最后与QD群体相聚.利用基因分化系数进行遗传变异来源估算,结果表明石鲽群体96.47%的遗传变异来源于群体内,群体间的遗传变异仅占3.53%,不同群体间的遗传相似度较大.     

Bayesian analysis of allozyme markers indicates a single genetic population of kingklip Genypterus capensis off South Africa

Kingklip Genypterus capensis inhabit deep continental shelf and slope waters off southern Africa and are now largely harvested as a bycatch in trawl and longline fisheries for Cape hakes Merluccius spp. Regional differences in growth, vertebral count and otolith morphology indicate that kingklip may consist of three stocks. In this study, allozyme markers are used to test whether the two southern stocks, the putative Cape and South Coast stocks, are genetically distinguishable. Standard electrophoretic methods detected Mendelian polymorphisms at six (23%) of 26 enzyme-encoding loci in five samples extending from about 29°S on the West Coast to about 25°E on the South Coast. No deviations from Hardy-Weinberg proportions were observed in the samples or in the pooled sample. Gene diversities (heterozygosity) ranged from H_S = 0.039 to 0.042 among samples, and averaged 0.041. Genetic differentiation among localities was remarkably small, F_ST = 0.003, and did not differ significantly from zero. A Bayesian analysis of population structure, based on detecting departures from Hardy-Weinberg proportions and gametic phase equilibria between loci, indicated with a high likelihood that the five samples were drawn from a single genetic population. In combination, these results are consistent with the occurrence of a single population off South Africa. However, the most conservative management strategy is to set harvest quotas individually on the Cape and South Coast stocks.     

Genetic heterogeneity in populations of the Mediterranean shore crab, Carcinus aestuarii (Decapoda,Portunidae), from the Venice Lagoon

Heterogeneity in genetic composition among recruits, mostly due to a large variance in reproductive success mediated by oceanographic processes, has been reported for marine species but is less understood in coastal lagoons' organisms. Temporal genetic variation in natural populations of the Mediterranean shore crab Carcinus aestuarii was quantified over a multi-year sample. A total of 486 adult crabs were collected at eight different sites of the Venice Lagoon during the period 2005–2007 and screened for genetic variation using 11 microsatellite loci. Two additional samples (N = 115) from neighbouring sites, located approximately 100 km North and South to the Venice Lagoon, were included for the sake of comparison. Our results show significant differences in allelic frequencies at the micro-geographic scale of the Venice Lagoon, observed between sites of collection, typologies of habitat, and areas with different class of ecological risk or pattern of hemocyanin expression. However, this pattern was not constant between years, with significant differences observed mainly in 2005 and 2006, but not in 2007. Our results indicate significant temporal differences suggesting the existence of dynamic processes that act on the genetic pool of this species. Although natural selection and gene flow might play a role, we suggest that genetic drift linked to large variation in the reproductive success of individuals is the most probable scenario to explain the local genetic patterns of differentiation in the Mediterranean shore crab. Our study, by providing the first evidence for the existence of genetic differences in this species at the micro-geographic scale, suggests that a better comprehension of the link between reproduction, recruitment and oceanography is critical to understand how colonization and maintenance of genetic variation is achieved in ephemeral and vulnerable environments such as coastal lagoons.     

Genetic structure of the common sole (Solea solea) in the Bay of Biscay: Nurseries as units of selection?

Coastal and estuarine areas in the Bay of Biscay are essential nursery habitats for sole (Solea solea). Using three intronic markers, we investigated patterns of genetic differentiation among cohorts of juveniles (0+, 1+, sub-adults) inhabiting four major nurseries in the Vilaine and Loire river estuaries, the Pertuis d'Antioche strait, and the Gironde estuary. Genetic differentiation was not significant for the age 0- and 1-groups, but was among sub-adult samples (θ_WC = 0.017; P = 0.031). Hierarchical analyses of genotypic frequencies by nested multivariate ANOVA using redundancy analysis indicated that northern nurseries of Loire and Vilaine rivers had different patterns of genotypic variation with age than southern nurseries of the Pertuis d'Antioche and Gironde estuaries (F = 1.36; P = 0.007). Bay of Biscay nurseries appeared to be spatially structured between a southern and a northern group that exhibit different genotypic distributions. Genetic drift and gene flow explains this pattern poorly, but a statistical test (P < 0.035) suggested that the metallothionein (MT) locus may be significantly impacted by selection in southern estuaries. As metallothionein is involved in heavy-metal detoxification, and southern nurseries are known to be affected by cadmium, this indicates a possible correlation between observed genotypic distributions and some nursery habitat features in sole. Some selective patterns might also be proposed to explain patterns of linkage disequilibrium observed at amylase loci in three of the four nurseries.     

马氏珠母贝家系遗传结构的微卫星分析

利用6个微卫星位点研究了9个马氏珠母贝(Pinctada fucata)养殖家系的遗传结构,为后续育种工作提供了指导。从60对引物中筛选出6对能够稳定扩增且多态性较好的引物。6个微卫星位点在9个家系中共检测到17个等位基因,每个位点等位基因数(A)为2~4个,平均2.833个。6个位点在9个家系上的有效等位基因数(Ne)平均值为2.030~2.632,平均杂合度观测值(Ho)为0.4306~0.6354,平均杂合度期望值(He)为0.4380~0.5962。家系间遗传分化系数(FST)为0.0749。采用NJ法进行聚类分析显示,9个家系分为明显的两支,根据遗传距离计算结果, F7和F9家系之间的遗传距离最近, F11与F12家系遗传距离最远。结果表明,9个家系的遗传多样性和遗传分化处于中等水平,可以考虑F11与F12家系杂交,以期获得较好的杂种优势, F11自交培育出遗传稳定的优良家系。     

Relationships between sediments and tissue contamination and allozymic patterns in Hediste diversicolor (Polychaeta Nereididae) in the Pialassa lagoons (north Adriatic Sea)

An increasing number of investigations revealed that responses of populations to environmental stressors may be genetically dependent. Occurrence of genetic adaptation to high concentrations of heavy metals in Hediste diversicolor local ecotypes has also been hypothesised. Analyses of allozymic patterns are a promising tool for the genetic monitoring of environmental stressors. The aim of this study was to investigate whether the allozymic patterns of H. diversicolor could be affected by levels of contamination. Five sites were investigated: four in a polluted brackish habitat, the Pialassa lagoons, and a reference site in the Lamone estuary. Total mercury was used as a tracer of the overall level of contamination. Concentrations of mercury were quantified in both sediments and H. diversicolor tissues by means of cold vapour atomic fluorescence spectroscopy. A gradient of mercury concentration in both sediments and H. diversicolor tissues was found. Moreover, significant differences in concentration of mercury were observed among and within sites. Allozyme electrophoresis was used to test the occurrence of genetic differentiation among H. diversicolor samples. A total of 19 putative gene loci were analysed. For six gene loci, significant variability in genotypic and allelic frequencies was detected. Patterns of differentiation, which could be related to the contamination levels, were found at loci LDH, PGI, SDH. Other loci (ALD, FH, HBDH) showed a significant genetic structuring among samples, not related to concentrations of mercury.