大黄鱼连续4代选育群体遗传多样性与遗传结构的微卫星分析

利用13个多态性微卫星位点分析了大黄鱼官井洋优快01品系F1到F44个选育世代的遗传结构与遗传多样性变化情况。结果显示,随着选育的进行,4个世代群体遗传多样性指标值渐次下降,F1到F413个微卫星位点的平均多态信息含量从0.638下降到0.524,平均等位基因数从5.462下降到4.308,平均观测杂合度从0.779下降到0.532,平均Shannon多样性指数从1.356下降为1.092。F1与其后各代遗传相似系数逐渐减小(从0.7194到0.5813),遗传距离逐渐增加,而相邻世代间的遗传相似性逐步升高,遗传分化指数(FST)渐次变小(F1～F2为0.0619,F2～F3为0.0511,F3～F4则为0.0475)。随着选育的进行,微卫星位点LYC0002和LYC0054等位基因频率有规律地发生变化,推测其可能与选育性状存在遗传上的相关。结果表明,经过连续4代的选育,部分不利基因遭到淘汰,选育群体的遗传基础逐步得到纯化,基因型逐渐趋向纯合、稳定,经进一步的选育可望获得较稳定的品系。     

乌鳢群体遗传多样性和遗传结构分析

为了解乌鳢群体遗传变异规律,本研究对8个群体共212个个体的mtDNA控制区全序列进行群体遗传多样性、遗传结构和群体历史动态分析。结果显示,乌鳢控制区全序列长度为907 bp,乌鳢群体单倍型多样性水平变化较大,中国黄河及以北的乌鳢群体单倍型多样性水平比淮河和长江等南方群体相对较低,所有群体表现出较低的核苷酸多样性水平(h0.5%)。基于单倍型构建的系统发育树和群体聚类树结果均未显示出与地理位置相对应的谱系结构。单倍型网络图显示存在多个主单倍型。遗传结构分析显示,不同水系间存在显著的遗传差异,相同水系间遗传差异较小。群体历史动态分析显示,中国乌鳢所有群体有效种群数量在中更新世晚期到晚更新世0.222—0.050百万年出现了一次较明显的快速增长,之后在晚更新世末次冰期0.050—0.010百万年出现了有效种群下降,伴随着全新世到来,在0.010百万年之后,乌鳢群体又发生了一次较小的有效种群增长。洞庭湖群体则发生一次有效种群的快速增长,增长时间大约在0.160百万年。研究表明,青藏高原隆起后,东亚季风在中国南北方气候的差异和秦岭山脉屏障对季风的阻断作用加大了这一差异,可能对乌鳢群体的遗传多样度差异造成一定影响。乌鳢群体不存在显著的谱系结构可能与乌鳢遗传分化时间较短有关,但是地理隔离等因素导致了不同水系间确切的遗传差异。第四纪更新世气候的波动,尤其是中更新世间冰期气候的转暖、末次盛冰期的降温和冰后期全新世的到来可能对乌鳢群体的数量和栖息地的扩缩起着重要影响。     

大竹蛏5 个野生群体遗传多样性的微卫星分析

运用微卫星标记对大竹蛏(Solen grandis)辽宁丹东(DD)、河北秦皇岛(QHD)、山东日照(RZ)、江苏吕四(LS)和广西北海(BH)近海5个不同地理野生群体共计150个样品进行了遗传多样性分析。结果表明:14个位点多态信息含量范围为0.696~0.853,均呈现高度多态性,每个位点检测到的等位基因数8~22个,共检测到199个等位基因,平均等位基因数为14.2,等位基因丰富度为11.05,5个群体的期望杂合度分别为0.769(DD),0.791(QHD),0.826(RZ),0.815(LS),0.785(BH),观察杂合度分别为0.837(DD),0.812(QHD),0.875(RZ),0.809(LS),0.858(BH),表明各群体处于较高的遗传多样性水平。Hardy-Weinberg平衡检验显示,仅有Sg16位点在丹东群体显著偏离平衡,其余位点在5个群体均正常,表明各群体遗传较稳定,处于平衡状态。5个群体间的遗传距离在0.141 2~0.340 9,DD和QHD的遗传距离最小,DD和BH的遗传距离最大;基于Da遗传距离构建的UPGMA聚类树显示,距离相邻的DD、QHD渤海湾群体聚为一支,RZ、LS黄海群体聚为另一支,最后与南海群体BH聚在一起,聚类结果与地理位置密切联系,基于贝叶斯遗传聚类得到了相同的结果。分析群体间的Fst值可知,两两群体间的Fst值在0.039 1~0.094 7,群体间产生了中等程度的遗传分化,并且达到了极显著水平(P=0.000 1)。由此可见,中国沿海各地理群体野生大竹蛏种质遗传多样性较为丰富,但不同群体间存在显著的遗传分化,故各增殖放流海区应当加强对放流苗种及繁殖亲本的种质检测,防止异地繁养等人为因素对大竹蛏各野生种群遗传结构造成破坏。     

清水江鲤基于微卫星标记和形态指标的遗传分析

利用12个微卫星标记对鲤(Cyprinus carpio)的4个野生群体 [清水江鲤、太湖鲤、黄河鲤(C.carpio haematopterus)和黑龙江鲤(C.carpio amurensis)] 和2个选育群体 [福瑞鲤(C.carpio var. FFRC)和松浦镜鲤(C. carpio var. specularis 'Song-pu')] 共208尾个体进行遗传分析。结果显示,12个位点共检测到341个等位基因,平均等位基因数为28.67,其中1个位点(HLJ1127)检测到正向选择压力;选育群体的遗传多样性参数普遍低于野生群体,其中松浦镜鲤群体的各项参数均值最低(N_a=6.82,H_o=0.54,PIC=0.50),清水江鲤群体的各项参数均值最高(N_a=21.25,H_o=0.80,PIC=0.91);分子方差分析显示,整体遗传变异主要来自群体内,但群体间呈极显著遗传分化(P<0.01);基于群体Nei's遗传距离的UPGMA聚类树和PCoA分析表明,鲤4个野生群体间遗传距离较近,而与2个人工选育群体间遗传距离较远;基于个体遗传结构及PCoA分析显示部分野生个体遗传结构比较混杂,而选育个体的遗传结构则相对单一。研究表明,中国鲤野生资源具有较高的遗传多态性,而人工选育群体维持着较纯的遗传种质。     

尼罗罗非鱼人工驯养、选育群体遗传多样性及瓶颈效应

人工驯养和选育是家养动物适应性进化的主要动力之一,中国大陆尼罗罗非鱼引进群体经历了长期的人工驯养和选育,是研究鱼类在驯养、选育条件下适应性进化的良好材料。本实验以尼罗罗非鱼1个埃及土著群体为对照组,以中国大陆具有代表性的尼罗罗非鱼5个驯养群体和4个选育群体为实验组,采用12个多态性微卫星位点分析了驯养群体和选育群体的遗传多样性和瓶颈效应。结果显示,土著群体、驯养群体和选育群体平均每个位点的有效等位基因数(AE)分别为5.433、5.113~6.515和3.239~6.734,期望杂合度(HE)分别为0.812、0.796~0.859和0.657~0.858,多态信息含量(PIC)分别为0.768、0.753~0.819和0.601~0.818,近交系数(FIS)分别为0.323、0.166~0.342和0.249~0.314。LSDt检验结果显示,受人工选择的群体(驯养群体、选育群体)与土著群体间遗传多样性水平(AE和HE)无差异,3个驯养群体(EGY群体、WY群体和GD群体)的遗传多样性水平(HE)显著高于1个选育群体(XJF群体)。瓶颈效应分析显示,尼罗罗非鱼土著群体、驯养群体和选育群体在历史上都曾发生过群体缩小的现象。其中,土著群体、2个驯养群体(WY、EGY)、2个选育群体(JNM、XJF)在近期可能经历过遗传瓶颈,其他群体在新的突变和基因流的作用下,群体规模可能已恢复。有效群体大小分析显示,土著群体、驯养群体和选育群体的有效群体数量分别为177、29~117(平均值为57.4)和84~123(平均值为102.8)。本研究结果不仅为尼罗罗非鱼驯养群体的持续利用和选育群体的进一步遗传改良提供了有价值的信息,而且为鱼类在驯养和选育条件下群体遗传结构和种群动态研究提供了新的参考依据。     

Microsatellite analysis of genetic diversity and genetic structure of the Chinese freshwater mussel Solenaia carinata (Bivalvia: Unionidae)

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日本沼虾微卫星引物筛选及群体遗传多样性分析

采用(CT)15、(CA)15两种生物素标记及磁珠富集法构建了太湖日本沼虾基因组微卫星富集文库。120个微卫星克隆中83个位点的核心序列重复在10个以上,长度介于191～580bp,平均为311bp;重复类型中,(CA/GT)n及(AG/TC)n分别占59.09%及20.45%,还检测到(GC)n、(AAG)n、(ACAA)n和(GGCAGA)n等17种类型,包括25.75%完美型、20.45%非完美型和53.80%复合性。用其中12条微卫星引物扩增吴江、滨湖、宜兴和吴兴4个群体240个个体的基因组DNA,各位点表现出较高的遗传多样性,除Mni86、Mni103和Mni114外,其它群体位点大都表现出显著的杂合子缺失,4个群体的遗传分化为低度分化(FST<0.05),4个群体中97.58%遗传变异存在于群体内,仅有2.42%的遗传变异来自于群体之间;吴江和吴兴群体亲缘关系最近,与宜兴和滨湖群体亲缘关系渐远。     

中国达氏鳇野生群体和两个养殖群体的线粒体遗传多样性分析

达氏鳇(Huso dauricus)是黑龙江流域土著鲟,近几十年来野生资源急剧下降,被确定为濒危物种之一。本研究采用线粒体DNA的Cyt b基因和D-loop区域的多态性信息评估了黑龙江抚远段野生群体、北京房山国家级鲟鱼原种场的保种群体及浙江衢州国家级鲟鱼良种场的繁殖群体等3个达氏鳇群体的遗传多样性水平。所有测试个体在Cyt b基因位点的核苷酸水平上没有检测到多样性,均具有相同的Cyt b单倍型,而在D-loop区域中发现了9种单倍型,对于D-loop区,单倍型多样性(H_d)达到0.593,但核苷酸多样性(π)仅为0.00213。在8尾野生个体中检测到6种D-loop单倍型,2个养殖群体共计58尾个体共计检出5种D-loop单倍型个体。分析结果显示:野生达氏鳇群体遗传多样性极低,历史上可能经历过严重的遗传瓶颈,同时达氏鳇人工繁殖过程中每批次可能只有极少个体参与了繁殖。此外,基于Cyt b基因部分序列的分析结果提示,达氏鳇与其他太平洋鲟类的亲缘关系较近,而与欧鳇(Huso huso)关系较远,传统上鳇属(Huso)的分类地位得不到有效的分子生物学数据支持。     

Genetic structure of Orbicella faveolata population reveals high connectivity among a marine protected area and Varadero Reef in the Colombian Caribbean

1. The protection of ecosystems with high diversity, such as coral reefs, is not an approach that guarantees their conservation. Thus, maintaining connectivity among coral populations over the long term is a strategy that should be adopted in order to protect diversity and ecological processes. Although coral reefs in Colombia are highly diverse, the population genetics baseline data of keystone species such as Orbicella faveolata are limited.
2. To provide current information about the connectivity between populations of O. faveolata in the Colombian Caribbean, the genetic diversity and distribution of O. faveolata was evaluated with analysis of molecular variance (AMOVA), principal components, and migration estimations using 113 genets distributed in six populations. Also, a genetic structure analysis that included the available data for the Caribbean population was conducted, seeking to understand how the Colombian populations relate to the broader region.
3. According to the global fixation index (F_ST) for the Colombian Caribbean, there was no genetic structure (F_ST = 0.002). Discriminant analysis of principal components (DAPC) showed that Corales del Rosario Archipelago (CR), Este de Isla Fuerte (EIF), San Bernardo Archipelago (SB), and Varadero Reef (VR) were grouped with Oeste de Isla Fuerte (OIF), with Bushnell (BS) representing the most divergent cluster. Migration analysis showed relatively high migration from VR to CR and SB, highlighting the importance of VR as a genetic reservoir for the region. Structure analysis showed that the Colombian population presented a specific genetic identity (F_ST = 0.254), suggesting that the Colombian Caribbean population could be a peripheral population that contributes significantly to genetic variation and is connected through a complex connectivity process.
4. In conclusion, the estimated genetic connectivity reflects the influence of sea surface dynamics over the interpopulation exchange dynamics and the role of protected and nonprotected coral reef in the Colombian Caribbean. Thus, the Colombian Caribbean population of O. faveolata could be relevant to the conservation of genetic diversity on a larger scale.

     

Isolation and characterization of koi herpesvirus (KHV) from Indonesia: identification of a new genetic lineage

Koi herpesvirus (KHV) is the aetiological agent of an emerging disease (KHVD) associated with mass mortalities in koi and common carp and reported from at least 30 countries. We report the first isolation of KHV from koi and common carp in Indonesia and initial characterization of the isolates. Clinical signs, histopathology and virion morphology are similar to those of isolates from other countries. Phylogenetic analyses using the thymidine kinase gene amplified from each isolate and from carp tissue samples collected from KHVD outbreaks throughout Indonesia indicated that the Indonesian isolates are more closely related to the Asian than the European KHV lineage. Sequence analysis of two other variable regions between ORF29 and ORF31 (marker I) and near the start of ORF 133 (marker II) indicated that all Indonesian isolates displayed a marker I allele (I(++)) previously identified only in isolates of the Asian lineage. However, in the marker II region, all Indonesian isolates displayed the II(-) allele, which has been reported previously only amongst isolates of the European lineage, and nine of these displayed a mixed genotype (II(+)II(-)). The I(++)II(-) genotype has not been reported previously and appears to represent a new intermediate lineage that may have emerged in Indonesia.     

Do dams increase genetic diversity in brown trout (Salmo trutta)? Microgeographic differentiation in a fragmented river

Abstract – Local genetic differentiation may potentially arise in recently fragmented populations. Brown trout is a polytypic species exhibiting substantial genetic differentiation, which may evolve in few generations. Movement (semi‐)barriers in rivers may cause fragmentation, isolation and genetic differentiation in fish. In the Måna River (28 km) flowing from the alpine Lake Møsvatn to the boreal Lake Tinnsjø, construction of four hydropower dams during the period 1906–1957 have fragmented the previously (since last Ice Age) continuous wild resident brown trout population. Samples from the two lakes (N = 40) and six sites in the river (N = 30) isolated at different times were analysed at nine microsatellite loci. All populations showed substantial genetic variation (mean number of alleles per locus 5.3–8.9, observed heterozygosity 0.57–0.65 per population, overall F_st = 0.032). Pairwise multilocus F_st estimates indicated no significant differentiation between populations in the two lakes, and no or little differentiation in the lower river (F_st = 0.0035–0.0091). The microgeographic differentiation among wild resident trout at these sites was less than expected based on similar previous studies. However, results from the upper river, in particular the site immediately below the Lake Møsvatn outlet and dam, indicated isolation (F_st > 0.035). Calculation of genetic distances and assignment tests corroborated these results, as did a significant correlation between years of isolation (since dam construction) and F_st. The population structuring is most likely a result of fragmentation by dams, which has increased overall genetic diversity. This increased local differentiation may be caused by natural selection, but more likely by genetic drift in small, recently fragmented populations. Increased local genetic diversity by genetic drift does not justify conservation measures aiming at preserving genetic diversity.     

基于SSR-seq技术评估中间球海胆多代选育群体和普通养殖群体的遗传多样性

为明确不同选育群体中间球海胆的遗传多样性和遗传结构,利用SSR-seq技术和15个微卫星位点,对1个家系选育群体(FP)、1个群体选育群体(IP)和1个未经选育的普通养殖群体(CP)的遗传多样性及遗传结构进行了分析。结果显示,15个微卫星位点共检测出112个等位基因,FP、IP、CP 3个群体的平均观测等位基因数(N_a)分别为5.077、5.133和6.133个,平均有效等位基因(N_e)分别为2.816、2.873和3.638个,平均观测杂合度(H_o)分别为0.522、0.441和0.501,平均期望杂合度(H_e)分别为0.595、0.599和0.667,平均多态性信息含量(PIC)分别为0.546、0.543和0.623。家系选育群体(FP) H_e与H_o的差值(0.073)低于IP (0.158)和CP (0.166),平均固定指数(F)(0.115)低于IP (0.248)和CP (0.246)。3个群体间遗传分化系数(F_st)介...     

High microsatellite genetic variability of the stone loach, Barbatula barbatula, in anthropogenically disturbed watercourses

Abstract  Genetic variation within and among stone loach, Barbatula barbatula L., populations inhabiting anthropogenically degraded watercourses in Flanders (northern part of Belgium) was assessed using five microsatellite markers. High levels of genetic diversity were observed at all sampling sites, (MNA: 6.2–11.2; H _O: 0.64–0.75; H _E: 0.67–0.85). Estimates of the effective population size varied between 1535 and 3021 individuals and there were no indications of recent severe bottlenecks. Significant genetic differentiation was observed among sites belonging to different river systems and drainage basins. These results suggest human activities, such as pollution and river engineering, have not impacted significantly on genetic variability in the stone loach populations investigated. It is possible that this lack of genetic erosion may be attributed to species-specific characteristics such as pollution tolerance and ecological flexibility.     

结合线粒体D-loop序列和SSR标记对宝石鲈养殖群体遗传多样性的分析

采用线粒体D-loop序列及SSR标记分析广东4个宝石鲈(Scortum barcoo)养殖群体的遗传多样性。D-loop序列分析显示,长度为598bp的D-loop片段上有14个多态性位点,共定义8个单倍型。4个群体单倍型多样性指数、核苷酸多样性指数分别为0.0000~0.5435、0.00000~0.00176,表明4个群体遗传多样性水平均较低。利用11个SSR位点分析显示,4个群体期望杂合度(He)=0.396~0.516、PIC=0.320~0.420,说明各群体遗传多样性处于中低水平。虽然二种方法获得的遗传分化指数FST上存在差异,分别为0.4035(D-loop)和0.0445(SSR),但二种分析结果均显示4个广东群体的遗传多样性较低、亲缘关系较近,因此有必要引进原种以丰富国内养殖群体的遗传多样性。     

津鲢与长江鲢遗传多样性分析

随机选取津鲢和长江鲢各36尾,用16个微卫星标记进行遗传多样性分析。结果显示:16个微卫星位点在2个群体中共检测到等位基因105个,基因型241种,每个位点等位基因数3～15个,平均6.6个,基因型4～37种,平均15.1种。2个鲢群体平均观察杂合度(Ho)分别为0.5393和0.5392,平均期望杂合度(He)分别为0.5887和0.5762,结果表明该2个鲢群体遗传多样性水平较高。2个鲢群体平均多态信息含量(PIC)分别为0.5602和0.54,固定系数(FIS)表明这2个鲢群体表现为杂合子缺乏(FIS0)。2个鲢群体之间的遗传距离为0.0566,平均遗传分化系数(Fst)值为0.021,说明仅有2.1%的遗传变异来源于群体间。     

Genetic connectivity and phylogeography of the night shark (Carcharhinus signatus) in the western Atlantic Ocean: Implications for conservation management

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The loss of genetic diversity in Dabry's sturgeon (Acipenser dabryanus,Dumeril) as revealed by DNA fingerprinting

• 1. Dabry's sturgeon, a large, long‐lived migratory fish is endemic to the Yangtze River. Over‐fishing and habitat destruction have caused large‐scale declines in natural stocks in the last two decades.
• 2. Examining patterns of genetic diversity has become an integral component of many management plans for endangered species. DNA fingerprinting was applied to detect genetic diversity in Dabry's sturgeon collected in 1958–1959, 1980–1981 and 1998–1999.
• 3. Studies on direct genetic parameters (genetic variability, hypervariable loci and heterozygosity) and indirect parameters (band‐sharing coefficient and allelic frequency) showed that the continuous decline in wild populations has caused the loss of genetic diversity in present‐day sturgeon.
• 4. The present‐day populations have the lowest genetic variability; thus, effective management is needed to preserve genetic diversity.
• 5. A conservation strategy is urgently required, comprising artificial rearing facilities coupled with breeding management plans.

Copyright © 2003 John Wiley & Sons, Ltd.     

黄颡鱼群体遗传变异分析

为了解中国黄颡鱼群体遗传变异规律,基于线粒体DNA控制区片段对本研究采集到的5个群体和文献收集的4个群体共258尾黄颡鱼进行群体遗传多样性、遗传结构、基因交流和群体历史动态分析。结果显示,在长度为413 bp的控制区片段上,9个群体的单倍型多样度在0.336±0.095~0.700±0.078之间,核苷酸多样度为0.087%±0.096%~0.258%±0.208%。基于所有单倍型构建的系统发育树结果显示不存在明显的谱系结构。单倍型网络图显示存在两个主单倍型。遗传结构分析显示不同水系间存在显著的遗传结构差异,其中洪泽湖和射阳河群体遗传结构位置不确定。群体历史动态分析表明,黄颡鱼群体存在群体扩张事件,扩张时间发生在末次间冰期时期。研究表明,9个群体遗传多样性呈现中—低等水平;射阳河和洪泽湖群体与长江和黄河水系在历史上尤其在黄河夺淮期间存在广泛的基因交流,导致其遗传结构位置不确定。黄颡鱼有效种群数量变化与第四纪冰期—间冰期气候波动有一定关系,中更新世气候转型后的末次间冰期升温可能导致了黄颡鱼群体扩张。     

An assessment of genetic diversity in wild and captive populations of endangered Japanese bitterling <Emphasis Type="Italic">Tanakia tanago</Emphasis> (Cyprinidae) using amplified fragment length polymorphism (AFLP) markers

ABSTRACT:   The Japanese bitterling Tanakia tanago (Cyprinidae) is on the verge of extinction in the wild, placing great importance on captive breeding programs for current conservation of the species. However, the loss of genetic diversity during captive breeding is an ongoing matter of concern. Since some captive populations have been almost monomorphic in mitochondrial DNA (mtDNA), this hampers assessments of their genetic diversity during captive breeding. To more accurately assess their genetic diversity, one wild and three captive populations were examined using amplified fragment length polymorphism (AFLP) markers. Estimates of average heterozygosity and nucleotide diversity ranged 0.0479–0.1920 and 0.0023–0.0088, respectively, enabling comparison of genetic diversity among the wild and captive populations, and among year-classes of captive populations. Significant differences in numbers of amplified fragments and proportions of polymorphic fragments were observed among year-classes of all populations. The indices of genetic diversity calculated from AFLP seemed to be, however, less sensitive to weak bottlenecks. No continuous decrease in genetic diversity in nuclear DNA was detected in presently captive populations. This supports the possibility of re-introduction of the captive populations into the original habitats, although survival and reproductive ability in the wild must be taken into consideration.     

A new map of the tiger shark (Galeocerdo cuvier) genetic population structure in the western Atlantic Ocean: Hypothesis of an equatorial convergence centre

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