图们江水系绒螯蟹的形态差异与遗传混杂

为了研究图们江水系中华绒螯蟹的种群现状,以图们江水系绒螯蟹为研究材料,将我国黄河和辽河水系的中华绒螯蟹(Eriocheir sinensis),以及日本本土的日本绒螯蟹(Eriocheir japonicus)作为参照对象,应用三种形态多元分析方法与STRUCTURE聚类分析方法,对它们的32个外部形态性状进行分析。判别分析显示,图们江群体的判别准确率最低(83.30%);主成分分析显示图们江群体的12个差异最大的表型性状均位于中华绒螯蟹与日本绒螯蟹之间,呈现为中间类型;传统聚类显示图们江水系绒螯蟹与日本绒螯蟹的形态差异最小;STRUCTURE聚类显示,图们江有60%的个体聚入日本绒螯蟹群体,而只有10%的个体聚入中华绒螯蟹群体,其余30%个体为中华绒螯蟹与日本绒螯蟹的中间类型。形态研究结果表明,图们江水系绒螯蟹为中华绒螯蟹与日本绒螯蟹分布的重叠区与混杂区,但其形态偏向日本绒螯蟹。     

中华绒螯蟹长江、黄河和辽河水系野生和养殖群体的遗传多样性

中华绒螯蟹(Eriocheirsinensis)是中国最重要的淡水养殖蟹类,广泛分布于东亚地区,养殖区域主要集中在长江、黄河和辽河流域。本研究基于线粒体DNAD-loop区评估辽河野生群体(LW)和养殖群体(LC)、黄河野生群体(HW)和养殖群体(HC)及长江野生群体(YW)和养殖群体(YC)的遗传多样性和种群结构。结果显示:(1)用于本研究的D-loop基因片段长度为477 bp,共包含234个变异位点和131个简约信息位点, 6个群体的262个个体中共有110个单倍型,包括90个独有单倍型和20个共享单倍型;(2)6个种群的单倍型多样性指数(Hd)范围为0.88889～0.96522,核苷酸多样性指数(π)范围为0.00887～0.01602,养殖和野生群体遗传多样性水平依次为:HCYCLC及HWLWYW;(3)6个群体的遗传距离(Da)范围为0.0119～0.0173,不论是养殖群体还是野生群体,辽河群体和长江群体之间的遗传距离均最小,且6个群体间遗传分化指数FST为0.12938。对6个群体进行中性检验显示Tajima’s D和Fu’s Fs的值均为负值。综上,基于线粒体D-loop基因的研究结果表明,三水系养殖和野生群体均具有较高的遗传多样性,且辽河和长江水系中华绒螯蟹的亲缘关系相对较近,该研究为中华绒螯蟹的种质资源评估、保护和开发提供了参考。     

Low genetic variability in an endangered population of fiddler crab <Emphasis Type="Italic">Uca arcuata</Emphasis> on Okinawajima Island: analysis of mitochondrial DNA

ABSTRACT:   Restriction fragment length polymorphism analysis was performed on polymerase chain reaction-amplified DNA fragments containing the D-loop , ND2 , and CO I genes of fiddler crab Uca arcuata mitochondrial DNA. In total, 316 individuals from six populations in Japan and two populations in Taiwan were analyzed using five restriction endonucleases ( Afa I, Bcn I, Cfr 13I, Hae III and Hin fI), yielding 85 haplotypes. Samples were taken from Nakagusuku Bay, Okinawajima Island, which is the only known distribution of U. arcuata in the Ryukyu Archipelago. The Okinawajima Island population is isolated geographically from others and showed a marked low genetic variability ( h  = 0.2539, π  = 0.0005) and significant differentiation from other population samples in haplotype composition. We suggest that a substantial decrease in the genetic variability of the Okinawajima Island population was caused by genetic drift under the conditions of small population size and low gene flow from other populations. It is important to conserve the intertidal zone in Nakagusuku Bay for the maintenance of this endangered population.     

ISSR标记在河蟹种质检测中的应用

将ISSR分子标记技术应用于天然水域河蟹(Eriocheir japonica)的遗传检测,结合来自母系的线粒体DNA分子标记鉴别,并通过对部分样本的细胞色素b基因变异位点测定,对ISSR标记检测结果做进一步分析和评价。对PCR/RFLP检测发现的2个携带中华亚种单元型的湛江青年河样本与来自鸭绿江、长江、闽江和南流江样本的ISSR检测与聚类分析表明:1)来自4个水系的河蟹聚为2大支,与中国大陆绒螯蟹的2个亚种分支相对应,即E.japonica sinensis和E.japonica hepuensis;2)来自南方水系湛江青年河的2个携带中华亚种单元型个体聚在北方的中华亚种(E.j.sinenesis)分支中。进一步对这2个样本的细胞色素b基因全序列测定分析表明,它们分别来源于中华亚种的ES2和ES15支系。提示ISSR标记可以提供天然水域河蟹种质混杂状况的核DNA方面的证据。同时表明,ISSR标记可以为河蟹种质资源的遗传研究提供有用的信息。[中国水产科学,2007,14(1):46-51]     

欧、美中华绒螯蟹源于中国长江水系中华绒螯蟹的遗传证据

用包括Z2和Opp17两个10碱基随机引物在内的10个引物,对来自荷兰斯科克莱（Skodely)、美国加州圣何塞（San Jose)的中华绒螯蟹群体与中国长江水系中华绒螯蟹群体进行RAPD遗传比较分析。结果发现：（1）中华绒螯蟹群体特有的Z2引物扩增的700bp标记带（Z2^700bp）,在荷兰与美国2个中华绒螯蟹群体中同时出现,而不出现日本绒螯蟹南流江种群中特有的880bp标记带（Z2^880bp）,表明欧洲、美国中华绒螯蟹与中国中华绒螯蟹为同种Eriocheir sinensis,而非日本绒螯蟹Eriocheir japonicus;（2）Opp17引物扩增的947bp片段在中国长江、荷兰及美国3个中华绒螯蟹群体内的出现频率均达100%。结合Z2引物扩增结果,欧洲与美国中华绒螯蟹群体极可能是从中国长江水系中华绒螯蟹引入繁衍的。     

Population structure of Japanese spiny lobster Panulirus japonicus inferred by nucleotide sequence analysis of mitochondrial COI gene

ABSTRACT:   The mitochondrial DNA sequence for the COI region was determined for 73 Panulirus japonicus individuals collected at three sample locations in Japan. Fifty-one haplotypes, including 69 polymorphic sites without insertion and deletion, were detected, of which three dominant haplotypes were shared among the three sample locations. The nucleotide and haplotype diversities at the three sample locations were in the ranges of 0.009–0.010 and 0.959–0.990, respectively. No significant population subdivision was detected among the sample locations based on the F _st value and net nucleotide substitution rate, and analysis of the molecular variance model ( amova ). There was no characteristic geographic distribution pattern for these haplotypes. These results indicate there is no genetic differentiation between P. japonicus populations in Japan, and support the hypothesis that benthic individuals of P. japonicus are sustained from a population of a common pool of long-period phyllosoma larvae through long-distance larval transport within the Kuroshio Subgyre.     

中国大陆沿海六水系绒螯蟹_中华绒_省略__群体亲缘关系_生化遗传差异分析

采用聚丙烯酰胺凝胶电泳分析比较了分布于我国大陆沿海 6个主要水系的两种绒螯蟹 (中华绒螯蟹、日本绒螯蟹 )群体间的生化遗传差异。结果发现 ,(1)辽河、黄河、长江和瓯江中华绒螯蟹群体间的生化遗传差异不显著 ,四个群体间的遗传距离D =0 .0 0 0 6～ 0 .0 0 53,属种内群体间差异。 (2 )珠江和南流江日本绒螯蟹群体间的生化遗传差异也不显著 ,两个群体间遗传距离D=0 .0 0 0 7,尚未达到亚种分化水平。 (3)辽河、黄河、长江和瓯江中华绒螯蟹与珠江和南流江日本绒螯蟹在SOD同工酶表型上有明显差异 ,SOD - 2位点仅在辽河、黄河、长江和瓯江中华绒螯蟹中表达 ,而SOD - 4位点仅在珠江和南流江日本绒螯蟹中表达 ,这些可作为中华绒螯蟹与日本绒螯蟹群体区分的生化遗传标志。辽河、黄河、长江和瓯江中华绒螯蟹群体同珠江和南流江日本绒螯蟹群体间的遗传距离较大 ,D =0 .0 72 1～ 0 .0 74 7。 (4 )聚类分析表明 ,在我国大陆沿海水系分布的绒螯蟹可分为两大类群 ,辽河、黄河、长江和瓯江水系的绒螯蟹属北方类群 ,即中华绒螯蟹 ;珠江和南流江的绒螯蟹属南方类群 ,即日本绒螯蟹。     

Limited genetic structure among broad-scale regions for two commercially harvested,tropical deep-water snappers in New Caledonia

We estimated the spatial population connectivity of Etelis carbunculus and E. coruscans based on measures of population genetic structure using the mitochondrial DNA control region. We collected samples from three areas separated by a minimum of 200 km around New Caledonia. We identified two separate genetic groups for E. carbunculus and a single group for E. coruscans. There were many singleton haplotypes distributed among geographic regions, indicating minimal spatial differentiation in genetic structure between regions for each species, although one of the genetic groups for E. carbunculus was in only two of the three regions. Conservation and management directives should consider both species as a single genetic stock among these three widespread regions. Our results provide evidence that stock structure might be less variable in the South Pacific than reported elsewhere, possibly indicating that trans-boundary management may be required in this region for deep-water snapper fisheries. Further refinement (including taxonomy) is required for E. carbunculus to characterize the different lineages observed.     

Genetic population structure of the deep-sea whelk Buccinum tsubai in the Japan Sea

ABSTRACT:   The purpose of the present paper was to examine the genetic population structure of Buccinum tsubai in the Japan Sea. Mitochondrial 16SrRNA gene sequence analysis was performed with specimens from various stations in the Japan Sea. Mitochondrial lineages were clearly patterned geographically in four separate areas: the Hokkaido area, the Yamagata–Toyama area, the Yamato Bank area, and the San'in area. The main distribution depth range of B. tsubai is between 200 m and 1000 m isodepths, and the horizontal distance between the 200 m and 1000 m isodepth lines represents the specific spatial scale of the habitat (SSSH). These four areas were separated either by the complete discontinuity of the SSSH area or by its narrow spatial extension. Genetic distances between the main haplotypes of each area were calculated as Jukes–Cantor distances, the value of which ranged between 0.012 and 0.017. This value seemed to be unrelated to the geographic distance. There was no tendency for clustering according to depth. In future, the morphological characters of the four lineages of B. tsubai should be compared in detail in order to elucidate significant genetic differences among them.     

Identification of currently cultivated Porphyra species by PCR-RFLP analysis

ABSTRACT:   Porphyra yezoensis and P. tenera are the representative species of the marine crop Porphyra (nori) in Japan. Since the two species are extremely similar to each other in morphology, nori breeders have tentatively classified many strains of cultivated Porphyra into the two species without strict species identification. In order to facilitate rapid and reliable identification of the many strains of Porphyra currently cultivated in various Japanese regions, 24 conchocelis strains were examined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis using the plastid RuBisCo spacer and the nuclear internal transcribed spacer regions. From the results, all the strains were identified as P. yezoensis f.  narawaensis , and P. tenera was not detected. Hence, it was confirmed that most of the Porphyra strains currently cultivated in Japan are P. yezoensis f.  narawaensis , and that intensive selective breeding has led to a reduced genetic diversity in the stock used for nori cultivation.     

我国蟹类土池育苗技术研究进展

蟹类养殖产业已成为我国水产养殖的支柱产业之一。中华绒螯蟹(Eriocheir sinensis,以下简称河蟹)、三疣梭子蟹(Portunus trituberculatus,以下简称梭子蟹)、拟穴青蟹(Scylla paramamosain,以下简称青蟹)是我国主要的养殖蟹类。目前3种蟹类土池育苗技术发展不平衡,河蟹、梭子蟹土池育苗技术较为成熟,已经普及推广并在养殖生产中发挥重要作用;青蟹土池育苗尚未有成功的报道。本文综述了我国河蟹、梭子蟹、青蟹苗种生产技术,尤其是土池育苗技术的研究进展,并对河蟹、梭子蟹土池育苗关键技术进行分析,旨在探讨蟹类土池育苗技术应用于青蟹土池育苗的可行性。     

基于CO I和Cyt b序列的丹江口水库鲢群体遗传结构特征分析

有效的人工增殖放流应监控放流群体和野生群体的遗传结构特征,以避免放流群体对天然群体遗传多样性的负面影响。本研究利用线粒体CO I和Cyt b基因分析了丹江口鲢库区、鲢亲本和鲢子代3 个群体的遗传结构特征。分析结果显示,104 条646 bp线粒体CO I序列中共检测到多态位点15 个,简约信息位点5 个,单一变异位点10 个,定义了7 个单倍型,单倍型多样性为0.544~0.676,核苷酸多样性为0.00221~0.00254;103 条1058 bp线粒体Cyt b序列中共检测到多态位点19 个,简约信息位点13 个,单一变异位点6 个,定义了14 个单倍型,单倍型多样性为0.609~0.714,核苷酸多样性为0.00262~0.00424,总体上处于较高单倍型多样性和较低核苷酸多样性。CO I和Cyt b序列遗传距离、遗传分化指数以及基因流分析结果显示,群体间遗传距离为0.002（CO I）、0.003~0.004（Cyt b）,总的遗传分化指数为-0.00468（P>0.05）（CO I）、0.03180（P>0.05）（Cyt b）,差异均不显著,群体间基因流为14.69～41.47（CO I）、5.49～40.47（Cyt b）,分子方差分析（AMOVA）结果表明群体的遗传变异主要来自群体内。单倍型聚类关系树表明,3 个鲢群体间均存在共享单倍型,不同地理群体间单倍型散乱分布于各支,未形成地理群体的聚集。以上分析结果显示,3 个鲢群体间不存在明显的遗传分化,表明增殖放流鲢群体与丹江口库区野生群体的遗传多样性和遗传结构相近,可开展增殖放流。本研究结果可为丹江口鱼类增殖站鲢群体的增殖放流提供科学依据。     

中华绒螯蟹大规格亲蟹ES-P2017家系规模化构建

基于中华绒螯蟹“长江1号”核心群体、苏南养殖群体、苏北养殖群体、长江野生群体和洪泽湖野生群体中选择大规格亲蟹,规模化构建ES-P2017家系,成功定向构建全同胞家系96个,家系构建成功率96.00%,选择56个发育良好的家系进行育苗,最终获得40个家系大眼幼体。     

Identification of alfonsino, Beryx mollis and B. splendens collected in Japan, based on the mitochondrial cytochrome b gene, and their comparison with those collected in New Caledonia

ABSTRACT:   The sequences spanning 307 bp of the mitochondrial cytochrome b gene were determined for 45 sepcimens of Beryx splendens and seven specimens of B. mollis collected in Japan, resulting in identification of 11 and three haplotypes in the two species, respectively. The parsimony tree was constructed from the determined sequences and those registered into the GenBank database as species A and W of B. splendens collected in New Caledonia, featuring with two clades. The first clade comprised species W from New Caledonia and B. mollis in the present study, whereas the second one contained species A from New Caledonia and B. splendens in the present study. These results demonstrate a large geographic distribution for both B. splendens and B. mollis. Some of the haplotypes found in Japan were identical to those of New Caledonia for both B. mollis and B. splendens , suggesting levels of gene flow at the trans -oceanic scale.     

Genetic and phenotypic comparison of Nocardia seriolae isolated from fish in Japan

The phenotypic and genetic characterizations of 58 isolates of the fish pathogen Nocardia seriolae , from amberjack, Seriolae dumerili , yellowtail, Seriola quinqueradiata , Japanese flounder, Paralichthys olivaceus , and chub mackerel, Scomber japonicus, in Japan from 1970–2005, were examined to investigate the epidemiological relationship between isolates. The phenotypic and genetic characterizations were determined by α-glucosidase activity and biased sinusoidal field gel electrophoresis (BSFGE) analysis, respectively. There was no α-glucosidase activity in strains isolated from 2000–05 ( n  = 50) with a few exceptions ( n  = 3), while all strains isolated from 1970–90 ( n  = 8) were positive. In BSFGE analysis, digestions with restriction enzymes Xba  I and Ase  I produced 15 and 16 restriction patterns, respectively. All restriction patterns obtained from 50 strains isolated during 2000–05 were unrelated to those obtained from eight strains isolated during 1970–90, with the exception of two strains isolated during recent outbreaks. Based on the phenotypic and genetic characterizations, recent outbreaks of nocardiosis in Japan are suggested to be epidemiologically unrelated to earlier outbreaks in Japan. Although a low genetic relationship was observed in the restriction pattern between recent and earlier isolates, identity was confirmed between these groups of isolates because five representative strains showed 99.9% homology with N. seriolae ATCC43993^T in the 16S rRNA sequence.     

蟹类原肌球蛋白基因的克隆与表达

原肌球蛋白是甲壳类动物的主要过敏原之一。本文通过分子生物学技术方法，分别克隆得到锯缘青蟹(Scylla serrata)、中华绒鳌蟹(Eriocheir sinensis)和三疣梭子蟹(Portunus trituberculatus)的原肌球蛋白基因序列。测序结果表明，三个基因的序列长度均为855 bp，编码284个氨基酸残基，三者氨基酸序列同源性为99.3%。三种蟹的原肌球蛋白基因序列与GenBank中其他甲壳类动物的原肌球蛋白具有很高的同源性。将锯缘青蟹的原肌球蛋白基因与pGEX-4T-3载体连接后，经IPTG诱导得到分子量约为61 kDa的融合表达蛋白。通过与甲壳类过敏患者血清的免疫印迹反应，证实融合表达的原肌球蛋白具有过敏原性，表明原肌球蛋白是蟹类的主要过敏原之一。该融合蛋白有望用于甲壳类食物过敏诊断试剂的开发与应用。     

白洋淀中华绒螯蟹种质资源形态判别与评价

对中华绒螯蟹外部形态32个测量点进行测量．利用形态判别法对白洋淀的中华绒螯蟹种质资源分析评价．建立了白洋淀中华绒鳌蟹形态判别式。结果表明．白洋淀河蟹种群属于北方蟹的辽河蟹．判别准确率达100％。结果有利于白洋淀中华绒螯蟹均质种群的维持，建议应加强白洋淀中华绒鳌蟹资源的管理。     

Genetic diversity and population history of the migratory catfishes Pangasianodon hypophthalmus and Pangasius bocourti in the Cambodian Mekong River

ABSTRACT:   Polymerase chain reaction (PCR) restriction fragment length polymorphism analysis of mitochondrial DNA was applied to the genetic structure and evolutionary history of the more ancestral Pangasianodon hypophthalmus ( n  = 82), and the recently speciated catfish Pangasius bocourti ( n  = 90) from the Cambodian Mekong River. Both pangasiids were characterized by a lack of genetic population structure that may result from high levels of contemporary gene flow. Genetic diversity was lower in P. hypophthalmus than in P. bocourti . However, a different evolutionary history was inferred for both species based on genealogical and demographic analyses (mismatch analysis, Tajima's D- and Fu's F _S-tests). The genetic profile of the more ancestral P. hypophthalmus shows indications of a recent population bottleneck, whereas the recently speciated P. bocourti shows signatures of historical population expansion. This study stresses the importance of preserving the migration routes in the Cambodian Mekong basin in order to maintain the genetic diversity and long-term integrity of both species.     

七里海中华绒螯蟹遗传多样性的RAPD和ISSR分析

利用RAPD-PCR及ISSR-PCR两种分子标记技术,分析了七里海中华绒螯蟹第6代繁育群体的遗传多样性。用10个多态性高的RAPD引物对24个七里海中华绒螯蟹样本个体进行扩增,共检测出107个不同的扩增位点,扩增片段大小为300~2300bp。其中多态位点总数为66个,平均多态位点比例为61.32%,群体内香农-维纳多样性值为0.1714。用8个多态性高的ISSR引物对24个个体共检测到106个位点,扩增片段为200~1500bp。其中多态性位点79个,平均多态位点百分率为74.53%,群体内香农-维纳多样性值为0.1778。结果显示,该群体的多态位点比例和香农-维纳多样性值均较大,说明其有较丰富的遗传多样性,同时也说明RAPD和ISSR技术用于七里海中华绒螯蟹核基因组的遗传多样性分析,具有较高的检出率和灵敏度。     

Genetic identification of native populations of fluvial white-spotted charr <Emphasis Type="Italic">Salvelinus leucomaenis</Emphasis> in the upper Tone River drainage

ABSTRACT:   Stocking of exogenous, hatchery-reared white-spotted charr Salvelinus leucomaenis has been conducted throughout much of their range in Honshu Island, Japan, to increase angling opportunities. Although the native charr populations are thought to have declined because of hybridization with introduced fish, their distribution and genetic status have been uncertain. Fine population structures of charr in the upper Tone River drainage were examined using mitochondrial DNA and microsatellite analyses so as to clarify the presence of native populations. One common mtDNA haplotype was detected in all populations in the Ohashi River and Watarase River, and four and one tributary populations were monomorphic for such haplotypes, respectively. However, several haplotypes, considered to have originated from stocked hatchery fish, were observed in the stocked and the remaining populations. Judging from the genetic integrity over a fine geographic scale, the former were considered as indicative of native populations and the latter as admixtures with hatchery fish. Comparisons of genetic diversity, deviations from the Hardy–Weinberg equilibrium, principal component analysis, and relatedness estimations based on microsatellite DNA can also provide evidence for distinguishing native populations from those influenced by hatchery fish.