凡纳滨对虾（Litopenaeus vannamei）不同世代养殖群体的遗传多样性分析

采用微卫星位点对凡纳滨对虾的海南群体（进口亲虾繁育的第1世代：G1）、山东和饶平群体（G2）、湛江2和湛江3群体（G3）、湛江1和上海群体（G4）共4个世代7个养殖群体的遗传多样性进行了分析。从21对微卫星引物中筛选出12对有效扩增引物,对7个群体共280个个体进行扩增,得到10个多态位点,共获得等位基因数63个。各位点的等位基因数在2-15个之间,各群体的平均等位基因数在4．70—5．80之间。平均观测（期望）杂合度在0.36—0．43（0．53—0．65）之间。杂合子偏离指数（D）为负值,表明存在杂合子缺失现象。对7个群体、10个多态位点进行Hardy—Weinberng平衡检测,共有54个偏离平衡。对7个群体间的遗传分化水平进行检测,得到近交系数FIS在9个多态位点均为正值,两两群体间的固定指数FST值在0．149-O．375之间。FST显著性检验表明,各群体之间的遗传分化极显著（P〈0．01）。分子方差分析结果显示,大部分遗传变异（72．17％）来自于群体内,来自于群体间的遗传变异（27．83％）较小。凡纳滨对虾不同世代间遗传变异大,随着繁育世代的增加,遗传多样性降低。     

刺参（Apostichopus japonicus）野生及两代选育群体间遗传变异的微卫星标记研究

应用微卫星DNA技术对野生刺参和两代选育刺参群体遗传多样性进行了研究。9对微卫星引物在三个刺参群体中共扩增获得43个等位基因,每个微卫星座位检测到的等位基因数为2—7个。三个群体的观测杂合度的平均值分别为0．6556、0．6704和0．6148,多态信息含量平均值分别为0．6654、0．5929和0．5275。结果表明...     

水体Hg^2＋对中华绒螯蟹（Eriocheir sinensis）血淋巴抗氧化酶活力和丙二醛含量的影响

采用单因子实验设计方法,进行了水体汞（Hg^2＋）在不同暴露时间对中华绒螯蟹血淋巴超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPX）和过氧化氢酶（CAT）活力以及脂质过氧化产物丙二醛（MDA）含量的影响研究。结果表明,较低浓度Hg^2＋（0．01和0．05mg／L）处理组中华绒螯蟹血淋巴SOD、GPX、CAT活力在...     

长江、黄河和辽河水系中华绒螯蟹野生和养殖群体遗传变异的微卫星分析

我国中华绒螯蟹(以下简称河蟹)养殖主要集中在长江、黄河和辽河流域,经过多年的人工养殖、跨流域引种和增殖放流等活动可能会对其遗传特性造成一定的影响,因此本研究利用29个微卫星标记对3个水系的野生和养殖群体进行了遗传特征分析。结果表明:(1)6个群体均表现出较高的遗传杂合度水平(Ho=0.702—0.744),香农信息指数(I)显示6群体的遗传多样性水平依次为:长江野生长江养殖黄河野生辽河养殖黄河养殖辽河野生。(2)分子方差分析(AMOVA)结果显示,来源于种群内个体间和个体内部的变异分别为14.02%和85.88%,群体间的遗传分化处于较低水平(FST0.05);(3)瓶颈效应和遗传结构分析显示,所有群体近期均经历过有效种群的降低,且6种群内个体的遗传组成混杂。综上所述,三水系野生和养殖群体均具有较高的遗传多样性,长江和黄河野生群体的遗传多样性略高于养殖群体;三水系河蟹野生群体的遗传分化与地理距离具有一定的相关性,长江养殖群体、黄河野生及养殖群体间的遗传分化较小可能与其跨流域引种及养殖群体逃逸造成其种质混杂有关。     

大菱鲆微卫星标记在亲本后代中的分离分析

为对大菱鲆(Scophthalmus maximus)的养殖群体进行微卫星分析研究,作者用70个微卫星位点在大菱鲆亲本后代中的基因型分离方式进行检测与分析。结果显示,70个位点共产生了12种分离方式(♀×♂)。70个微卫星位点中45个位点符合孟德尔遗传定律(P≥0.05),有25个位点偏离了孟德尔遗传定律(P0.05)。平均等位基因数目为2.2,平均有效等位基因数为1.885。平均期望杂合度和观测杂合度分别为0.55和0.537。多态信息含量为0.182~0.699,平均值为0.361。研究证明这些标记大部分可以用于大菱鲆进一步的图谱构建,QTL定位和分子标记辅助育种研究。同时对群体的分析表明,该大菱鲆养殖群体遗传多样性水平较高,可用于进一步的良种繁育。     

日本绒螯蟹放流群体的遗传变异

利用水平电泳方法研究日本绒鳌蟹放流群体（新县的三面川和岛根县的高津川的遗传变异。计检测了15种同工酶,22个位点。两河川日本绒螫蟹的多态座位比例及平均杂合度）利力0．273和0．020,0．227和0．022,与其他种群基本一致。另外,两群体其他相近两自从群体的根井遗传距离为0．001。结果表明：到目前为止,尚未见到放流群体的遗传变异的明显减少。     

长江、黄河、辽河水系中华绒螯蟹(Eriocheir sinensis)野生和养殖群体遗传变异的微卫星分析*

我国中华绒螯蟹(以下简称河蟹)养殖主要集中在长江、黄河和辽河流域, 经过多年的人工养殖、跨流域引种和增殖放流等活动可能会对其遗传特性造成一定的影响,因此本研究利用29个微卫星标记对3个水系的野生和养殖群体进行了遗传特征分析。结果表明：(1) 29个SSR标记共检测到943个等位基因,各位点等位基因数在11-52之间, 平均为32.52个; 各位点的平均观测和期望杂合度分别为0.723和0.876,仅5个位点处于Hardy-Weinberg平衡中;各位点平均多态信息含量(PIC)为0.864,其中28个为高多态性位点(PIC>0.5)。(2) 6个群体均表现出较高的遗传杂合度水平(H_O=0.702-0.744);香农信息指数(I,介于2.566-2.661)显示6群体的遗传多样性水平依次为：长江野生>长江养殖>黄河野生>辽河养殖>黄河养殖>辽河野生。(3)方差分析(AMOVA)结果显示,来源于种群内个体间和个体内部的变异分别为14.02%和85.88%,群体遗传分化指数F_ST仅为0.0008-0.0050,群体间的遗传分化处于较低水平(F_ST<0.05);基于Nei’s遗传距离的UPGMA聚类结果显示,长江野生群体单独聚为一支, 其余5群体共同聚为大一支, 其中长江养殖群体、黄河野生群体与黄河养殖群体聚为一小支,辽河野生群体和养殖群体聚为另一小支。(4) 瓶颈效应分析显示,所有群体近期均经历过有效种群的降低。群体遗传结构分析显示显示,6种群内个体的遗传组成混杂,不能根据其遗传结构划分出最佳理论群体数。综上所述,三水系野生和养殖群体均具有较高的遗传多样性,长江和黄河野生群体的遗传多样性略高于养殖群体;三水系河蟹野生群体的遗传分化与地理距离具有一定的相关性,长江养殖群体、黄河野生及养殖群体间的遗传分化较小可能与其跨流域引种及养殖群体逃逸造成其种质混杂有关。     

泥蚶人工选育群体的微卫星分析

采用微卫星分子标记技术对人工选育的泥蚶(Tegillarca granosa)群体进行遗传多样性分析。结果表明:在12个标记位点中,每个位点的等位基因数为2~9个,观测杂合度平均范围0.4195~0.4725,期望杂合度平均范围为0.5700~0.6323。哈迪-温伯格平衡检验结果显示,在48个群体-位点组合中有32个群体-位点组合显著偏离平衡。Shannon遗传多样性指数为1.0608~1.2242。群体间的遗传分化系数Fst值从0.0347~0.4549,平均值0.1930;基因流Nm值为0.2996~6.9617,平均值2.3367;群体近交系数Fis范围0.0052~0.7135,平均值0.2468。结果表明,连续的人工选育过程,群体的遗传多样性仍然比较高,但选育过程对群体的遗传多样性和遗传分化有一定程度的影响。     

中华绒螯蟹与日本绒螯蟹线粒体COI基因片段的序列比较研究

以相应引物 PCR扩增了黄河口中华绒螯蟹线粒体细胞色素氧化酶 I亚基基因 (COI)片段 ,PCR产物经 T载体连接之后进行克隆、测序 ,得到 70 9bp的碱基序列 ,其 A,T,G,C含量分别为 34.4 1% ,2 7.93% ,2 0 .0 3%和 17.6 3%。并比较它与珠江流域中华绒螯蟹 COI序列和日本绒螯蟹 COI序列的差异 ,发现黄河口中华绒螯蟹与珠江流域中华绒螯蟹 COI序列完全相同 ,而与日本绒螯蟹差异非常明显 ,70 9或 6 5 8(不计引物 )位点中核苷酸差异数为 32 ,核苷酸差异率为 4 .5 1%或 4 .86 % (不计引物 ) ,其中 2 5个位点为转换 ,7个位点为颠换。作者倾向于支持存在中华绒螯蟹和日本绒螯蟹 ,或它们为同一种的两个地理亚种的观点     

花尾胡椒鲷养殖群体遗传多样性的等位酶研究

应用聚丙烯酰胺凝胶电泳（PAGE）技术对取自厦门火烧屿海水网箱的花尾胡椒鲷人工鱼养殖群体的遗传多样性进行等位酶分析。结果表明：所检测的16种同工酶的24个基因座位中，在XDH和GCDH基因座位上发现多态性，其平均多态位点比例为8．33％，基因座位有效等位基因的平均数Ne为1.083，平均杂合度的观测值Ho为0．00556，预期值He为0．00537,Hardy-Weinberg遗传偏离指数（D)为0．035。与其他鱼类相比较说明了其遗传多样性处于较低水平，文中对此进行了探讨并提出相应的种质资源管理保护措施。     

广东沿海杂色鲍养殖群体遗传多样性的微卫星分析

实验选取的7个杂色鲍(Haliotis diversicolor)养殖群体分别来自广东汕头、汕尾、惠东、湛江和徐闻,深圳野生驯养,利用7对微卫星引物对上述群体进行了遗传多样性和群体遗传结构的分析。结果显示:7个位点产生的等位基因数从7～13个不等,共检测到68个等位基因,平均每个位点的等位基因数为9.71个。有效等位基因数的范围为3.8～8.1,平均观测杂合度范围为0.422～0.906,平均期望杂合度范围为0.748～0.917。7个群体的多态信息含量变化范围为0.657～0.883,PIC值均在0.500以上,表现为高度多态性。与两个野生驯养的养殖群体相比,其他5个养殖群体均存在不同程度的遗传多样性降低。AMOVA分析显示,4.79%的遗传变异来自于群体之间,95.21%的变异来自于群体内的个体之间。研究表明,广东沿岸的杂色鲍具有较高的遗传多样性水平,7个群体之间呈现一定的遗传分化。     

Genetic architecture of three Turbinella pyrum varieties (Linnaeus, 1758) from the southeast coast of India

The genetic diversity and variation of three pseudo‐species of the commercially important Indian conch Turbinella pyrum from the southeast coast of India were studied. A total of 88 individuals was genotyped at 12 loci and differences in the genetic diversity among genetic groups were correlated with known population histories. The study identified 535 alleles and all the studied loci were highly polymorphic. The number of alleles ranged between four and 29 with a global mean of 14.861. The global means of the observed and expected heterozygosities were 0.774 and 0.891, respectively. The estimated within‐population inbreeding value (F_IS = 0.131) indicated a shortfall of heterozygosity in the populations. An analysis of molecular variance showed that 5% of the total variation was due to differences between genetic groups. The Bayesian approach revealed strong distinctiveness in the three genetic groups studied. Overall microsatellite analysis revealed less genetic diversity in all studied genetic groups and significant genetic variation among the three pseudo‐species. The study highlights the significant distinctiveness of the pseudo‐species of T. pyrum and the need for appropriate conservation efforts in order to prevent further losses of their genetic diversity.     

大刺鳅(Mastacembelus armatus)二、三、四碱基重复微卫星标记的筛选和特征分析

对大刺鳅(Mastacembelus armatus)进行简化基因组测序,开发二、三、四碱基重复类型的微卫星引物共105对(重复次数≥6),其中可稳定扩增的引物有50对(47.6%),通过多态性分析,发现有38个微卫星位点表现出多态性,从中随机选取三种重复类型微卫星各10对对北江群体进行遗传多样性分析,及各6对对澜沧江大刺鳅群体进行遗传多样性分析。结果显示,(1)在大刺鳅中二碱基重复微卫星位点筛选效率为50%,而三、四碱基重复的筛选效率分别为31.1%、35%。(2)北江群体的平均多态信息含量为0.649,比澜沧江群体(0.572)高,30个微卫星位点在北江群体中共检到235个等位基因,且93.3%的引物表现为高度多态水平,18个微卫星位点在澜沧江群体中共检到120个等位基因,66.7%的引物表现为高度多态水平。(3)二碱基重复的微卫星标记在两个野生群体检出的五项多态性指标高于三、四碱基重复。结果表明,二碱基重复的微卫星比三四碱基重复的微卫星具有更高的筛选效率和多态性,北江群体的遗传多样性比澜沧江群体高,本研究所筛选的引物可以应用于群体间遗传多样性分析和遗传图谱的构建,以及亲缘关系的鉴定等方面,为大刺鳅种质资源保护研究奠定分子基础。     

Genetic variability of half-smooth tongue sole Cynoglossus semilaevis populations using microsatellite markers

Ten highly variable microsatellite loci were performed in order to evaluate the genetic variability of wild and hatchery samples of half-smooth tongue sole Cynoglossus semilaevis. A group of 200 genotypes belonging to four wild samples, Laizhou (LZ), Weihai (WH), Qingdao (QD), Rizhao (RZ) and one hatchery sample, Mingbo (MB), were screened. All of the ten microsatellite loci screened in this study showed marked polymorphism. A total of 70 different alleles were observed over all loci. The number of alleles per locus ranged from 4.2 to 10.6. The average of expected and observed heterozygosity ranged from 0.67 to 0.82, and from 0.77 to 0.87, respectively. A total of 10, 16, 10, 11 and 5 unique alleles each were found in LZ, WH, QD, RZ and MB populations. The effective number of alleles varied from 2.87 for HSTS_7 to 6.83 for HSTS-h. The number of average genotypes ranged from 6.0 for HSTS_a to 15.4 for HSTS_h. As compared with the wild populations, the hatchery population, MB, showed significant genetic changes such as fewer alleles per locus (P <0.05), a smaller number of low frequency alleles (P <0.05), a small number of unique alleles and a small number of genotypes (P <0.05), all indicative of a reduction in genetic diversity.     

The influence of environmental variability of a coastal lagoon ecosystem on genetic diversity and structure of white seabream [Diplodus sargus (Linnaeus 1758)] populations

As coastal lagoons serve as nursery areas for some marine and estuarine fish, selective pressures of these brackish or hypersaline lagoons may influence the genetic structure of species and populations. We examined spatial and temporal genetic patterns at eight microsatellite loci in white seabream [Diplodus sargus (Linnaeus 1758)] recruits from the Mar Menor (Southeast Spain) and compared these loci with those in coastal populations from the open sea, observing a high degree of genetic diversity and spatio‐temporal genetic stability. However, the results suggest the presence of subpopulations or genetic substructures in the Mar Menor D. sargus population that could be interpreted as a homogeneous mixture of individuals from three differentiated subpopulations in the Mediterranean and evidence of the Wahlund effect. It also suggests that D. sargus adults return to their original spawning habitat, thus conserving the genetic differences among the respective populations over time. Overall, this study demonstrates the importance of the Mar Menor coastal lagoon as a nursery area for the conservation of genetic diversity of D. sargus populations.     

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…     

基于微卫星(SSR)分子标记的中国对虾渤海湾增殖亲虾群体及回捕群体的遗传多样性分析

Eight microsatellite markers were used to analyze genetic diversity, level of inbreeding, and effective population size of spawner and recaptured populations of Chinese shrimp(Fenneropenaeus chinensis) during stock enhancement in the Bohai Bay in 2013. A total of 254 and 238 alleles were identified in the spawner and recaptured populations, respectively, and the numbers of alleles(N_a) were 8–63 and 6–60, respectively. The numbers of effective alleles(N_e) were 2.52–21.60 and 2.67–20.72, respectively. The polymorphism information content ranged from 0.529 to 0.952. The observed heterozygosity(H_o) values(0.638–0.910 and 0.712–0.927) were lower than the expected heterozygosity(H_e) values(0.603–0.954 and 0.625–0.952), which indicated that the two populations possessed a rich genetic diversity. In 16 tests(2 populations×8 loci), 13 tests deviated from the HardyWeinberg equilibrium. F_(is) values were positive at seven loci and the inbreeding coefficients(F) of the two populations estimated by trio ML were 13.234% and 11.603%, suggesting that there was a relatively high degree of inbreeding. A certain level of inbreeding depression had occurred in the Chinese shrimp population. F_(st) values ranged from 0 to 0.059, with a mean of 0.028, displaying a low level of genetic differentiation in the two populations. Effective population sizes(3 060.2 and 3 842.8) were higher than the minimum number suggested for retaining the evolutionary potential to adapt to new environmental conditions. For enhancement activity in 2014,the ideal number of captured shrimp spawners should have ranged from 7 686 to 19 214 to maintain genetic diversity and effective population size. Further strategies to adjust the balance of economic cost, fishing effort and ideal number of shrimp spawners to maintain a satisfactory effective population size for ensuring the sustainability of Chinese shrimp are proposed.     

Parentage determination of black sea bream(Acanthopagrus schlegelii) for stock enhancement:effectiveness and loss of genetic variation

The stock enhancement programs for black sea bream Acanthopagrus schlegelii have been conducted in China for a fewyears.However,little information has been reported concerning the effectiveness and genetic effect of black sea bream stock enhancement.In order to detect the contribution of released individuals in Zhujiang River Estuary(ZRE) and Daya Bay(DB),six microsatellite markers were used to identify the hatchery-released individuals.In addition,this pedigree of hatchery populations(broodfish and hatchery-released offspring) was traced to detect the number of effective parents(N_e),the inbreeding coefficient and the decrease of genetic variability in the reproduction.The pedigree reconstruction showed that at least 69(out of 93) broodfish had offspring.The estimated N_e was 54.8,consequently the inbreeding coefficient was 0.91%.The genetic diversity of hatchery-released offspring was lower than that in that of broodfish(heterozygosity alleles,0.727-0.774),some alleles(number of alleles,61-69) and genetic variance were lost during reproduction.It was observed that wild samples had higher levels of genetic diversity compared with hatchery populations as well as recaptured samples in releasing area.A total of 128 hatchery-released black sea bream were identified among 487 recaptured samples in ZRE,while a total of 15 samples were identified among 96 samples in DB.In summary,there was a high survival of released fish.Nevertheless,the results provided evidence to consider a loss of genetic variation in hatcheryreleased stock and a negative genetic effect of the stock enhancement.     

许氏平鲉(Sebastes schlegeli)微卫星标记开发及野生、养殖群体遗传多样性分析

对许氏平鲉进行简化基因组测序,设计微卫星引物200对,可稳定扩增的引物190对,占95%。利用一个荣成野生群体对24个多态性较高的微卫星标记进行了评价,每个位点的等位基因数(Na)为2—21个,观测杂合度(Ho)为0.0417—0.9167,期望杂合度(He)为0.0278—0.9722,多态信息含量(PIC)为0.1948—0.9496,结果显示有20个微卫星位点为中高度多态。利用这些引物对荣成野生群体和烟台养殖群体的遗传多样性进行了比较分析,野生和养殖群体的平均等位基因数(Na)分别为8.5000、6.9583,有效等位基因数(Ne)的均值分别为4.5484、3.6365,期望杂合度(He)均值分别为0.6421、0.5840,多态信息含量均值(PIC)分别为0.6088、0.5490,平均香农-威纳指数均值为1.4605、1.2834,但F检验发现无显著差异,发现两个群体的遗传多样性都处于高度多态水平,但养殖群体遗传多样性水平低于野生群体。本研究结果说明许氏平鲉的人工繁育中,通过使用较大数量的亲本进行繁育可有效防止选育群体的遗传多样性降低,但人工定向选育对选育群体的遗传多样性也产生了一定的影响。Bonferroni校正后在两个群体中各有4个位点偏离Hardy-Weinberg平衡。本研究开发的微卫星标记为许氏平鲉遗传图谱构建、分子标记辅助育种等提供了更多标记选择,对野生和养殖群体遗传多样性分析为下一步的遗传育种提供参考。     

长牡蛎(Crassostrea gigas)野生与选育群体的微卫星遗传多样性分析

为了评估长牡蛎(Crassostrea gigas) 2个壳长性状(掌心形)快速生长选育群体(LY2-K4、LY2-K7)、1个壳高性状(速生型)快速生长选育群体(LY2-K11)和6个野生群体(QHD、LS、HD、ZH、WD、KTD)的遗传多样性和遗传结构, 用21对多态性丰富的微卫星引物对9个长牡蛎群体的269个个体进行了遗传分析。结果显示: 21个微卫星位点共检测出了460个等位基因(N_a),平均等位基因数为21.905; 21个微卫星位点的多态信息含量(PIC)均大于0.5, 具有高度遗传多态性; 选育群体LY2-K11的遗传多样性最低(N_a=13, I=2.128,H_e=0.831, PIC=0.825), 野生群体KTD的遗传多样性最高(N_a=29,I=3.112, H_e=0.941, PIC=0. 938); 189个群体位点组合有66%偏离哈代-温伯格平衡, 表明这些群体存在一定程度的杂合子缺失; 9个群体间的遗传分化指数(F_st)为0.012~0.064, 处于较低的遗传分化水平; AMOVA分析显示遗传变异主要来自于个体内; PCoA分析结果与UPGMA聚类树一致, LY2-K11群体单独聚为一类, QHD和HD群体聚为一类, 其他6个群体聚为一类。综上所述, 长牡蛎3个选育群体和6个野生群体遗传多样性均较高, 遗传分化水平较低; 选育群体LY2-K11多样性略有下降, 选育过程中应保证亲本的数量及质量, 防止因近交衰退造成遗传多样性降低, 苗种抗逆性变差。该结果将为长牡蛎新品种的选育和野生种质资源的保护提供科学指导。