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.     

Genetic variability analysis in five populations of the sea cucumber Stichopus (Apostichopus) japonicus from China,Russia, South Korea and Japan as revealed by microsatellite markers

The genetic structure of populations of the sea cucumber Stichopus (Apostichopus) japonicus was investigated using 10 microsatellite markers. In all, 152 individuals from five natural populations were collected from Aomori, Japan (JA and JR), Yosu, South Korea (KY), Dalian, China (CD) and Vladivostock, Russia (RV). A total of 145 alleles were found at 10 loci. The number of alleles per locus ranged from 9 at PSC03 to 20 at SCZ06, with an average of 14.5. Average H_o and H_e ranged from 0.260 (JR) to 0.434 (JA) and from 0.654 (RV) to 0.778 (KY), respectively. No significant differences at A, H_o and H_e were found, indicating similar genetic diversity in the five populations. A single allele was found at the PSC05 locus in the RV population. Of the 50 loci, 42 significantly deviated from Hardy–Weinberg equilibrium, all showing heterozygote deficiency. The genetic distances were all relatively great, ranging from 0.497 (between JA and KY) to 1.029 (between KY and JR). This suggests the five populations are genetically distinct. Cluster analysis indicated that JA, KY and CD form one branch and RV and JR another in the UPGMA tree. A hypothesis is proposed for the evolution of the Japanese red sea cucumbers and the genetic relationship among the populations.     

舟山近海棘头梅童鱼群体遗传多样性微卫星DNA分析

应用6对可在棘头梅童鱼（Collichthy lucidels）中扩增的大黄鱼（Pseudosciaena Crocea）微卫星引物对浙江舟山近海的棘头梅童鱼群体进行PCR扩增,变性聚丙烯酰胺凝胶电泳分析扩增产物.6对引物在群体中共扩增出46个等位基因,平均每个位点得到5.308 0个有效等位基因,各位点的PIC值为0.412 6~0.893 5（平均值0.670 8）,PC1C4和PC8F5为中度多态性位点,PC4H12、PC5E11、PC10F10及PC10G6为高度多态性位点,这些位点可以做为棘头梅童鱼群体遗传学研究的有效的分子遗传标记.6个位点的连锁分析显示,各位点间不存在明显的连锁关系.各位点在群体中的观测杂合度（Ho）为0.454 5~0.916 7（平均值0.659 1）,期望杂合度（He）为0.468 0~0.901 9（平均值0.699 7）,与其他海水鱼类比较,浙江近海的棘头梅童鱼群体遗传多样性偏低.对各位点进行Hardy-Weinberg平衡检测表明,PC4H12、PC10F10及PC10G6位点的等位基因频率偏离了平衡,综合现有的资源调查资料,遗传多样性的降低与近年来棘头梅童鱼资源的下降有关.     

Assessment by microsatellite analysis of genetic diversity and population structure of Enhalus acoroides from the coast of Khanh Hoa Province, Vietnam

Seagrass beds degraded significantly since the last century on both, global and local scale. The seagrass species Enhalus acoroides (Linnaeus f.) Royle is a common species found in almost all marine ecosystems including bays, lagoons and around offshore islands in tropical regions of the West Pacific. It was shown that genetic diversity is an essential indicator of the conditions of ecosystems. In the present study, microsatellite markers were used to assess the genetic diversity and population structure of six distinct seagrass beds along the coast of the Khanh Hoa Province, Vietnam. The results indicate that the genetic diversity of the populations in the open sea is higher than in the lagoon. Seagrass beds occurring in disturbed sites show reduced genetic diversity. The fixing index value (F_ST) depicts a relatively high genetic structure among populations. Structure analysis clusters the populations into open sea and lagoon populations and cluster analysis and AMOVA indicate a significant difference between the two groups. There are low but non-significant positive correlations between geographic and genetic distances. The different habitats of the open sea and the lagoon are probably responsible for forming two groups.     

Concordance of microsatellite and mitochondrial DNA markers in detecting genetic population structure in the boring giant clam Tridacna crocea across the Indo‐Malay Archipelago

Mitochondrial DNA (mtDNA) is a single, usually non‐recombining locus, and often uniparentally inherited. Therefore, its ability to reveal recent gene flow among populations is usually questioned. In this study, the genetic population structure of 16 populations of Tridacna crocea (n = 366) from the Indo‐Malay Archipelago (IMA) was examined with 10 microsatellite markers and compared to previous studies using mtDNA, in order to test if the revealed population structure was congruent between the two marker systems. The results showed that the genetic population structure revealed by the two marker systems was mostly congruent, with a high correlation between cytochrome c oxidase subunit I (COI) and microsatellites. The studied populations could be divided by both marker systems as follows: (i) Eastern Indian Ocean, (ii) Central IMA, and (iii) Western Pacific. Populations in the Central IMA showed high gene flow. However, populations in the Java Sea (Karimunjava, Pulau Seribu) were grouped into a separate cluster by mtDNA analysis, while this grouping was not detected by microsatellites. It was also noteworthy that there is obvious heterozygosity deficiency in most of the populations, which may be caused by null alleles, inbreeding or population expansion. Overall, these results indicate that the mitochondrial COI gene is applicable for population genetic analysis and precise recovery of connectivity patterns of giant clams. Therefore, the combination of mtDNA and nuclear DNA markers can lead to a more complete understanding of population genetics. Moreover, this study is expected to facilitate fully displaying the population genetic structure of giant clams combining with other researchers' results.     

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。     

Genetic diversity and population connectivity of the sea urchin Tripneustes gratilla along the South African coast

The collector sea urchin Tripneustes gratilla has been identified as a species with potential for aquaculture production in South Africa. The species’ roe is considered a culinary delicacy in Asia and Europe. However, T. gratilla remains genetically uncharacterised in South Africa. Therefore, the purpose of this study was to provide baseline genetic information consisting of estimates of genetic diversity and population stratification, which may aid in future sustainable use of this urchin resource. A total of 22 species-specific microsatellite markers were used for the genetic characterisation of T. gratilla samples from along the South African coast, at Haga Haga, Coffee Bay, Hibberdene, Ballito Bay and Sodwana Bay. A moderate level of genetic diversity was observed, with an average number of alleles of 7.89 and an average effective number of alleles of 6.57, as well as an average observed heterozygosity of 0.55. Population differentiation tests suggest that the geographically representative samples form part of a single, large interbreeding population, with a global F_st estimate of 0.02 (p > 0.05). This finding is likely explained by high levels of gene flow between these locations caused by extensive larval dispersal during the planktonic larval stage. The panmixia observed within these natural populations of T. gratilla indicate that they could be managed as a single genetic stock.     

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.     

漠斑牙鲆养殖群体微卫星遗传多样性的初步分析

为评估养殖过程对漠斑牙鲆遗传结构的影响, 本研究利用微卫星标记对漠斑牙鲆(Paralichthys lethostigma)山东胶南养殖群体(子一代)和江苏南通养殖群体(子三代)的遗传结构进行分析比较, 结果表明所使用的14 对引物中, 11 对引物在所选取的群体中具有多态性, 两个群体中共检测出了70 个等位基因(42 个有效等位基因), 总群体的平均观察杂合度(Ho)和期望杂合度(He)分别为0.36 和0.60。两个群体的Shannon’s 遗传多样性指数分别为1.18 和1.07, Nei’s 遗传多样性指数分别为0. 56 和0.51, 表明山东胶南群体的遗传多样性显著高于江苏南通养殖群体。两个养殖群体间的两群体间的遗传分化系数Fst 平均为0.1246, 遗传距离(Ds)为0.171, 遗传相似度为0.8432, 表明两个群体中度分化。由此可以得出, 在中国随着漠斑牙鲆累代养殖的开展, 其遗传多样性下降显著, 遗传结构已经出现了显著分化。因此, 在将来的繁育过程中应制定合理的育苗规划以维持漠斑牙鲆的多样性水平并保障产业可持续发展。     

Genetic variation of Chinese and Japanese wild Pacific abalone(Haliotis discus hannai) measured by microsatellite DNA markers

1Introduction ThePacificabaloneHaliotisdiscushannaiisan importantspeciesinthecoastalfisheriesofnorthern China,andisofhighcommercialvalue.Thepro ductionofcultureseedsfortheabalonebeganin Chinaintheearly1980s.Duringthelasttwodec ades,productionofabalonehasi…     

方斑东风螺3个选育世代遗传多样性和遗传结构的微卫星分析

采用12个多态性微卫星标记对方斑东风螺(Babylonia areolata)泰国选育系TF₄~TF₆连续3代选育群体的遗传多样性与遗传结构进行了分析。3个群体的多态信息含量分别为0.730、0.717和0.708,均高于0.5,表现出较高的多态性。平均等位基因数(A)、有效等位基因数(A_e)、观测杂合度(H_o)、期望杂合度(H_e)和Shannon’s信息指数(I)的变化范围分别为11.667~12.583、6.334~6.915、0.658~0.672、0.737~0.787和1.837~2.003,整体呈降低趋势,但差异并不显著(P>0.05)。分子方差分析结果显示,群体间的遗传变异仅占总变异的0.95%;群体间的遗传分化系数(F_st)介于0.00569~0.01324,处于低等分化水平(F_st<0.05)。主坐标分析和STRUCTURE分析结果显示,3个群体呈现出相似的遗传背景和遗传结构。以上研究结果表明,经过连续多代的人工选育,方斑东风螺选育群体保持着较高的遗传多样性水平,遗传变异和遗传分化水平较低,遗传结构趋于稳定,仍具有较高的遗传选育潜力。     

Spatio-temporal variations in reproductive patterns and population structure of Pasiphaea multidentata (Decapoda: Caridea) in the Blanes canyon and adjacent margin, North-western Mediterranean Sea

Pasiphaea multidentata is a deep‐water caridean shrimp fished in the Mediterranean in association with the commercially exploited red‐shrimp Aristeus antennatus. A previous study describes seasonality in the reproductive pattern of P. multidentata using external morphological parametres. This study assesses the spatio‐temporal variations in the population structure, sex ratio, ovary cycle and gametogenesis of P. multidentata from three different fishing grounds in the Blanes canyon and adjacent margin (North‐western Mediterranean) over an annual cycle. The oogenetic pattern of this species is typical of a caridean shrimp. There is a pool of previtellogenic oocytes at all times that develop from the periphery of the gonad towards the centre during maturation. Previtellogenic oocytes grow to approximately 200 μm before undergoing vitellogenesis. The vitellogenic oocytes are surrounded by a monolayer of accessory cells. The maximum size observed for a mature oocyte was 1420 μm. The oocyte‐size distribution confirmed the seasonal reproductive pattern of this species; in winter, the ovaries contained mainly previtellogenic oocytes, some of which start maturing in spring, resulting in a slightly bimodal distribution. In summer, the vitellogenic oocytes reach approximately 1000 μm and in late autumn the ovaries are fully mature and ready to spawn. There were no significant differences in the reproductive and population structure patterns of P. multidentata among the three sites, suggesting that the population’s distribution is not affected by the geomorphology of the area, in particular the presence of the canyon. The populations are dominated by females at all sites and all seasons, with the arrival of juveniles in spring. The seasonal variations in the reproduction and recruitment of P. multidentata and the lack of spatial segregation within the population are discussed in terms of the species’ known biology, the effects of canyons in energy supply to the deep‐sea floor and the relationships of this species with the red‐shrimp A. antennatus.     

皱纹盘鲍野生与养殖群体微卫星标记遗传变异研究

采用7个微卫星标记对辽宁大连、山东烟台、荣城、崂山和胶南5个皱纹盘鲍养殖群体,以及山东长岛、荣城、日本岩手、韩国仁川4个皱纹盘鲍野生群体进行了群体遗传多样性与遗传分化的研究.结果表明,从等位基因数与杂合度上分析,养殖群体(N=8.0～9.4,He=0.754～0.787)遗传多样性显著低于野生群体(N=11.3～15.0,He=0.821～0.866);等位基因频率分布揭示出野生群体中一些稀有等位基因在养殖群体中有所丢失;亲鲍选用数量少与性别比例不均衡可能是产生养殖群体中遗传多样性显著减少的原因.通过对等位基因频率比较分析,5个养殖群体之间以及养殖群体与野生群体之间观察到显著差异的Fst与Rst值,野生群体之间日本群体Fst与Rst值与其他3个野生群体差异显著,群体间存在遗传分化;养殖群体间遗传距离与地理距离不相关,可能是皱纹盘鲍育苗过程中亲鲍和苗种频繁交流所造成的结果;日本群体与其他3个野生群体间存在显著遗传分化与皱纹盘鲍短暂的浮游期与有限的扩散能力有关.     

Multiple paternity in the common long‐armed octopus Octopus minor (Sasaki, 1920) (Cephalopoda: Octopoda) as revealed by microsatellite DNA analysis

Multiple paternity was assessed in Octopus minor using microsatellite DNA markers. Octopus minor adults were captured in traps and kept in indoor cement ponds. The females spawned after several days. Ten broods (B1–B10), each containing 15 embryos and the maternal octopus, were maintained until the embryos reached the stage at which DNA could be extracted and used for genotyping. Multiple paternity was tested using five microsatellite DNA markers and the results proved the hypothesis that multiple paternity occurs in O. minor (observed in six out of the 10 broods). Analysis with GERUD software gave the minimum number of fathers and showed that four broods were sired by a single male, three were sired by two different males and three were sired by three different males. In brood B10, the number of fathers identified by the single‐locus method was one fewer than that by the GERUD method. Of the six multiple paternity broods, four (66.7%) showed significant skew from equal paternal contributions, which indicated that sperm competition and/or cryptic female choice may be important for post‐copulatory paternity bias in this species.     

The genetic diversity,individual relatedness and possible mating system of an isolated population of the Cyprinid species Megalobrama pellgrini in upper reaches of the Changjiang (Yangtze) River,China

Megalobrama pellegrini is a cyprinid fish endemic to upper reaches of the Changjiang(Yangtze)River,China,which is also an important economic species in the local area.In recent years,resources of this species have decreased sharply and its conservation has drawn great attention.In the present study,we collected 120 individuals from the Longxi River,a tributary isolated from the main channel of the Changjiang River,where M.pellegrini is still relatively abundant.Using two different molecular markers,mitochondrial cytochrome b(cyt b)gene,and nuclear microsatellite(simple sequence repeat,SSR),we analyzed the genetic diversity of this isolated population.The results show that sequence genetic diversity was low(Hd:0.290 and P i:0.000 77 for cyt b gene),while the SSR genetic diversity was high(Ho:0.824 4±0.147 2,He:0.823 5±0.145 1).Analysis indicated that this population had experienced a bottleneck,with inbreeding and small effective population size(around 50).Based on SSR data,relatedness analyzing revealed that the 120 samples were grouped into 10 completely independent clusters.It was inferred that the mating system of M.pellegrini was polygamy.We suggested that the low genetic diversity could be induced by the overfishing and inbreeding depression.Therefore,we suggested that the urgent conservation measures should be taken to control the overfi shing and give better conditions for the fish to grow and spawn,then to restore population size.     

Spatial and temporal variations in the population structure of the soft-shell clam Mya arenaria in the Pomeranian Bay (southern Baltic Sea)

The population structure of Mya arenaria has been investigated in the sublittoral zone of the Pomeranian Bay (southern Baltic Sea). Box-corer samples were collected during a 1.5-year period in 1993/94 to follow changes in size and age structure of the clam populations in different parts of the study area. Large spatial differences in population structure were found between the sheltered southwest of the Bay and the shallow and exposed Oder Bank in the centre. The stock of the Oder Bank consisted of two different clam types. A slow-growing cohort was assumed to be autochtonous on the Oder Bank, whereas a fast-growing one was assumed to have immigrated from the surrounding area. The contribution of the two cohorts to the total density varied seasonally. Because of bedload transport of clams, the contribution to the reduction of the clam stock by wintering sea ducks could not be quantified. In the southwest of the Pomeranian Bay erosion was of minor importance. High mortality rates during the first two years of life were assumed to be caused by predation. Mortality rates of older cohorts remained stable until old age. Variations in cohort strength were related to interannual differences in the reproductive success. A mild winter presumably lowers the reproductive success in the subsequent summer. Severe oxygen depletion in summer 1994 caused a strong reduction in the clam stock at stations deeper than 10 m.