我国沿海不同群体文蛤遗传差异RAPD分析

本文运用RAPD对辽宁、山东、江苏、福建、广东、广西等7个地理群体的文蛤进行了分析。从40条随机引物中,筛选出11个扩增效果好、条带清晰的随机引物进行扩增。结果显示,7群体内遗传多样性丰富,遗传相似度在0.7251~0.8972之间,依次排列为:辽宁营口(0.8972)>广东湛江(0.8756)>山东日照(0.8729)>山东潍坊(0.8643)>福建云霄(0.8245)>江苏启东(0.7653)>广西合浦(0.7251)。群体间遗传距离指数及用UPGMA和NJ法构建的分子系统树结果表明,山东日照与山东潍坊两群体的亲缘关系较近,广东湛江群体与其它群体亲缘关系相对较远。     

合浦珠母贝3个养殖群体遗传多样性的微卫星分析

采用微卫星分子标记对我国广东大亚湾、广西北海、海南三亚的合浦珠母贝养殖群体进行了遗传多样性分析.从49对引物中筛选出31对有效扩增引物,种群扩增获得9个多态位点,多态位点比例为29.03%.它们在3个群体共96个个体中产生了44个等位基因,平均每个位点产生4.889个.3个群体的平均期望杂合度分别为0.590、0.600、0.615;平均观察杂合度分别是0.393、0.425、0.325;平均多态信息含量PIC值分别是0.530、0.551、0.556.表明3个群体的遗传多样性处于较高水平.平均遗传偏离指数分别为-0.324、-0.336、-0.304;哈代-温伯格平衡检测发现,3个群体27个位点中21个位点偏离平衡状态.遗传分化和遗传距离分析表明,海南群体与广东群体之间的亲缘关系较近,而与广西群体的亲缘关系较远.     

青蛤北方3个群体遗传多样性分析

采用随机扩增多态性DNA(RAPD)技术对辽宁省大连庄河海区、锦州海区及山东省东营海区青蛤群体的遗传结构和遗传变异进行分析.15条随机引物共检测到123个位点(250～2000 bp),3个群体的青蛤最大遗传距离为0.0549,遗传相似性系数超过94%,群体分化不明显,未形成不同的地理种群.3个群体总的DNA多态位点百分率为94.2%,表明3个群体青蛤当前种质资源状况良好,遗传多样性水平较高.聚类分析显示辽宁锦州群体和山东东营群体优先聚类,亲缘关系较近.     

银鲳4个地理种群遗传多样性的初步研究

采用随机扩增多态DNA(RAPD)技术对取自海南(HN)、浙江(ZJ)、江苏(JS)、河北(HB)等近海海域的4个地理群体银鲳进行遗传多样性分析。从20个随机引物中筛选出10个,利用POPGEN Version 1.31软件对多态位点比例和遗传距离进行统计分析。共检测出总位点数56个,其中多态位点有39个,多态位点比例为69.64%。4个海域银鲳群体的Shannon多样性指数分别为0.123 2~0.158 7,Ne i基因多样性指数分别为0.077 6~0.100 4。JS群体与HN群体的遗传距离最远,为0.321 6;JS群体与HB群体的遗传距离最近,为0.231 6。结果表明,RAPD技术具有较高的灵敏性和多态性,是分析评估银鲳遗传多样性较为适宜的分子标记之一;JS群体保持着最高的遗传多样性,而HN群体多态性比例最低、ZJ群体Shannon多样性指数最低,应采取一些有效的保护措施,以避免HN群体遗传多样性的进一步丧失。     

日本沼虾三个野生群体亲缘关系分析

为研究我国野生日本沼虾洞庭湖、鄱阳湖、龙感湖3个不同地理群体之间的亲缘关系,利用微卫星(SSR)分子标记技术对上述3个群体进行了亲缘关系分析。从47对引物中筛选出11对引物对日本沼虾上述3个不同地理群体的基因组进行PCR扩增,共扩增90个条带,其中69个(占76.6 ％)为多态性条带。参照pBR322DNA/MspI Marker估算所扩增的条带大小,根据条带的位置确定其基因型,并利用GENEPOP32软件进行遗传距离和相似指数数据分析。结果发现:日本沼虾3个不同地理群体间遗传距离最大为0.196 5,最小为0.186 1,平均遗传距离为0.188 8。采用NTsys2.20聚类分析软件的UPGMA聚类法进行聚类分析的结果表明:龙感湖群体与鄱阳湖群体的亲缘关系最近,而与洞庭湖群体的亲缘关系最远。     

福建东山和广东阳江褐毛鲿养殖群体遗传多样性的AFLP分析

应用AFLP分子标记技术对福建东山和广东阳江褐毛鲿养殖群体进行了遗传多样性分析。采用6对选择性引物组合对2个群体60个个体进行扩增,共扩增出313个位点,多态位点85个。东山和阳江褐毛鲿养殖群体的多态位点比例、Nei’s基因多样性指数、Shannon遗传多样性指数分别为24.60%和25.56%、0.0795和0.0768、0.1210和0.1176,2个群体的遗传多样性均处于较低水平。遗传分化系数Gst及AMO-VA分析表明,2个群体的遗传变异主要来源于群体内个体间。UPGMA聚类图及PCA分析显示,群体间具有典型的地理特征,且出现了一定程度的遗传分化。     

基于线粒体 Cyt b 基因序列的洞庭湖河蚬遗传多样性分析

研究洞庭湖不同地理群体河蚬的遗传多样性和遗传结构,为其资源合理开发和有效保护提供参考。在6个样点共采集河蚬126只,以线粒体细胞色素b基因部分序列为分子标记,分析扩增与测序后基因序列的单倍型及多样性、核苷酸多样性,计算群体间遗传分化指数等。获得长531bp的序列片段,共检出5种单倍型,变异位点10个;平均单倍型多样性和核苷酸多样性分别为0.559和0.006。6个河蚬群体间的遗传距离为0.006~0.007;遗传分化指数和AMOVA分析显示,河蚬群体间遗传分化低,遗传变异主要来自群体个体内。最大似然法构建的系统进化树中,单倍型DTH1和DTH5聚为一支,DTH2、DTH3、DTH4聚为另一支。中性检验结果显示洞庭湖河蚬群体相对稳定,没有经历过瓶颈效应和突然的群体萎缩等历史事件。洞庭湖6个河蚬群体间不存在显著的遗传差异,可将其作为一个整体进行管理和保护。     

中国东北部山区细鳞鱼4个群体遗传多样性分析

采用微卫星法,利用11对微卫星引物对黑龙江省松花江流域、吉林省图们江支流红旗河流域、吉林省白山市鸭绿江流域及吉林省图们江支流密江流域随机采集的94尾细鳞鱼样本进行遗传多样性分析,探讨4个不同流域细鳞鱼群体的遗传多样性差异及其亲缘关系。结果显示,4个群体共检测到等位基因147个,各群体有效等位基因数为1.6541~2.4143,平均观测杂合度为0.3849~0.5186,平均期望杂合度为0.3488~0.5003,4个群体平均多态信息含量为0.3320~0.4704。亲缘关系分析显示,红旗河流域群体与密江流域群体遗传距离最小(0.0954),亲缘关系最近。松花江流域群体与其他3个群体的遗传距离较大,亲缘关系较远。群体间相似指数在0.7112以上,相似性较高。     

文蛤山东种群与江苏种群杂交及自繁子代的遗传差异分析

利用fAFLP技术研究了文蛤山东种群与江苏种群的杂交子代（SJ，JS）和自繁子代（SS，JJ）群体的遗传差异。3对荧光引物组合在4个F1群体96个个体中共扩增出255个位点，多态位点比例高达97.65%。Nei’s基因多样性和Shannon信息指数显示，4个群体的遗传多样性大小依次为SJ＞SS＞JS＞JJ，而且两杂交组的变异度都大于自繁组，说明杂交增加了文蛤种群的遗传多样性和变异程度。fAFLP扩增位点分析表明，SJ和JS的显性基因频率有所增加，并且正、反杂交群体在遗传结构上均表现出明显的偏母性特征。从群体间遗传相似性指数来看，JS与JJ间的遗传相似性系数最大（0.976 1），而与SS间的遗传相似性为0.945 0；SJ与SS间的遗传相似性系数为0.950 2，与JJ群体间为0.925 4；SJ群体与JS群体间的遗传相似性系数最小（0.907 5）。在依据群体间相对遗传距离构建的系统树上，JS与JJ间的亲缘关系最近，首先聚在一起，SJ与SS间亲缘关系较近，也能聚为一支，但明显可以看出正交组合SJ与其它3个群体间的亲缘关系较远。综合分析表明，文蛤地理种群杂交增加了遗传多样性和变异度，正、反交群体在遗传结构上都明显地偏向母本，正交组合SJ群体的遗传多样性最高、变异度最大，这与其在生长速度上表现出的杂种优势可能有直接关系。     

大泷六线鱼6个群体遗传多样性的微卫星分析

大泷六线鱼资源捕捞量逐渐加大,捕捞个体体型却逐渐减小,研究大泷六线鱼的遗传多样性,对提高在渔业养殖中种群质量具有重要意义。本研究利用7对微卫星分子标记对大泷六线鱼(Hexagrammos otakii)的6个地区群体(大连、丹东和青岛等地)进行了遗传多样性和遗传变异评估。结果表明,获得的等位基因数1~8,平均等位基因数6.29;有效等位基因数1.00~5.72,平均有效等位基因数4.71;各群体的杂合度观测值(Ho)为0.05~0.27;杂合度期望值(He)为0.44~0.75;多态信息含量(PIC)为0.47~0.69,说明这6个群体属于高度多态,遗传多样性水平较高。群体间遗传分化系数(F_(st))为0.27,表明6个大泷六线鱼群体之间存在明显的遗传分化。采用UPGMA法对6个大泷六线鱼群体进行聚类分析,发现大连和长海县大泷六线鱼群体亲缘关系较近且聚为一个分支,其他4个群体聚为另一分支。结合Hardy-Weinberg平衡检验分析,发现各群体表现出极显著的遗传不平衡,不符合Hardy-Weinberg平衡定律,表明6个群体大泷六线鱼遗传多样性较为丰富。综上说明,6个大泷六线鱼群体发生了较大的遗传分化,遗传多样性较为丰富。     

Suspension feeding and growth of ark shell <Emphasis Type="Italic">Anadara granosa</Emphasis>: comparison with ubiquitous species <Emphasis Type="Italic">Scapharca subcrenata</Emphasis>

The ark shell Anadara granosa is a species peculiar to the Ariake Sound. To determine why distribution of A. granosa in Japan is largely confined to this area, we examined feeding and growth of A. granosa as functions of environmental and biological variables. The results were compared with those of another ark shell Scapharca subcrenata, which is ubiquitous in Japan. Feeding experiments indicated that A. granosa is eurythermal and euryhaline, as is S. subcrenata, but is adapted to temperature slightly higher than S. subcrenata. Weight-specific clearance rate (CR) of A. granosa as a function of soft-body dry-weight (w) followed the power function of w (CR=2.7×w ^−0.37), with coefficient and exponent very close to those for S. subcrenata. Growth rate of A. granosa increased linearly with daily ration, similar to S. subcrenata. Thus, feeding and growth characteristics of A. granosa were comparable to those of S. subcrenata and no ‘peculiarities’ of the former were detected. Therefore, the factors that make A. granosa a species restricted to the sound are probably not directly related to feeding or growth characteristics.     

Genetic diversity and population history of Tanichthys albonubes (Teleostei: Cyprinidae): Implications for conservation

1. Tanichthys albonubes is a cyprinid fish of South China and North Vietnam. Although this species has been sold worldwide in the aquarium trade for more than a century, it is listed as a second‐class state‐protected animal in China and classified as ‘extinct in nature’ in the China Red Data Book.
2. To investigate the population history of T. albonubes and evaluate the genetic conditions among the wild and hatchery populations, mitochondrial genes (mtDNA, 2032 bp from the d‐loop and cyt b), nuclear genes (nuDNA, 2241 bp from RAG1 and ENC1) and 13 microsatellite loci were used to assess the genetic structure throughout the range of this species.
3. In total, 358 specimens were collected from three hatcheries and all known wild populations. This study found that the discordant population structure among these genetic markers was accounted for by differences in the effective population sizes. The results indicated that (1) the three hatchery stocks originated from a single source in China and that the hatchery stock in China maintained almost all the T. albonubes genetic diversity in the downstream reaches of the Pearl River; (2) the wild population near Baiyun Mountain originated from hatchery releases; (3) the population history reflects the complex geological history of South China; and (4) the habitat destruction and fragmentation that have resulted in small and isolated populations may have shaped the genetic structure of T. albonubes.
4. The low‐level genetic diversity of T. albonubes supports the need for conservation interventions. The geographically distinct genotypes indicate a need for the development of management strategies directed towards the conservation of localized populations. The genetic status of all populations, including the hatchery stocks, should be evaluated and monitored continuously.

     

基于线粒体Cyt b基因的虾夷扇贝群体遗传结构分析

本研究利用线粒体细胞色素b(Cyt b)基因标记对虾夷扇贝(Patinopecten yessoensis)6个群体(长岛底播增殖群、海洋岛底播增殖群、獐子岛底播增殖群、旅顺自然群、"獐子红"人工选育群以及日本青森陆奥湾群)进行种质状况评估,研究结果表明,6个群体120个个体的Cyt b基因序列共检测到20种单倍型,其中"獐子红"人工选育群单倍型最少,日本群体单倍型最为丰富,两者单倍型多样性指数分别为0.10000和0.88400。分子方差分析(AMOVA)发现,日本群体与中国群体间变异百分比为15.34%,明显高于作为一个基因池时群体间遗传变异水平;就中国组群而言,其83.41%的遗传变异来自于群体内部,组内群体间的变异只占1.52%,说明中国群体个体间的遗传变异远大于群体间的遗传变异。F_(st)分析显示,中国群体与日本群体之间发生了中等水平的遗传分化(0.07455~0.17895,Fst0.05)。以遗传距离矩阵构建群体间分子系统树(UPGMA树),拓扑结构图显示中国5个群体先聚为一支,再与日本群体聚为一支。由此可见,中国养殖区的虾夷扇贝与日本原产地群体间已出现明显的遗传分化,且虾夷扇贝中国群体的遗传多样性处于较低水平。本研究结果可为虾夷扇贝的种质资源保护与可持续开发利用提供理论依据。     

Genetic diversity and structure of Northern Sheatfish (Silurus soldatovi) assessed by newly developed and cross‐species microsatellites

Twenty‐five polymorphic microsatellite markers, including 13 newly isolated markers and 12 existing cross‐species markers, were used to assess the genetic variation of 141 individuals from three cultured stocks of northern sheatfish, Silurus soldatovi, in China. Samples were collected from Heilongjiang (H), Tongerpu (T) and Liaoning (L). Allelic richness, expected and observed heterozygosity among 25 loci were calculated, and the mean A_e, H_o and H_e were estimated in three stocks. In low polymorphic locus HLJcf19 and HLJcf38, most individuals in all three stocks were homozygous and had only one same allele, while only few heterozygous individuals had the other allele in one certain stock. And all the three stocks showed excess of heterozygotes, because of significant negative F_IS values. Genetic distance, UPGMA dendrogram, as well as structure analysis, all indicated that Stock H and L were closer among three stocks. The results revealed that though all stocks had no reduction in recent time, their effective population size were significantly lower to the ideal value.     

基于CMSY和BSM的东海区重要渔业种类资源评估

有限数据方法(data-limited method)可结合少量易获得数据和相关生物学信息对渔业资源状况、生物学参考点以及生物量等进行评估,已经成为全球区域性渔业管理组织和资源评估学者的关注热点。本研究采用基于渔获量的最大可持续渔获量(catch-based maximum sustainable yield,CMSY)和基于贝叶斯状态空间的Schaefer产量模型(Bayesian Schaefer production model,BSM)评估了东海区19个重要经济种类的资源状况,并提出了基于最大可持续渔获量(maximum sustainable yield,MSY)的渔业管理建议。结果显示,19个种类中有1个种类衰竭,3个种类严重衰退,5个种类过度捕捞,5个种类轻度过度捕捞,5个种类健康。种群状态长期评估结果表明,处于生物可持续水平的鱼类种群占比已由1980年的95%下降至2019年的26%。同时对CMSY和BSM方法的结果进行了比较,整合单位捕捞努力量渔获量(catch per unit effort,CPUE)数据的BSM方法导致了置信区间较宽,并调节了生物量轨迹的变化形态...     

我国五大湖日本沼虾线粒体COI基因部分片段序列比较

对我国五大湖日本沼虾100个野生个体的线粒体细胞色素氧化酶亚基I（COI）部分序列进行了测定和分析，经比对获得578bp核苷酸片段，发现49个变异位点，得到35个单倍型，包括7个共享单倍型，各群体都具有较好的单倍型多态性和核苷酸多态性，其中鄱阳湖群体遗传多样性相对最高。AMOVA分析表明，五群体间总遗传分化系数F_st＝0.31873 （P<0.05）， 群体间具有较高的遗传分化。MEGA3.1软件计算五群体的Kimura 2-paramter遗传距离，洞庭湖群体和巢湖群体之间的遗传距离最远为0.0191，巢湖群体和洪泽湖群体之间的遗传距离最近为0.0051。以同属胖掌沼虾（Macrobrachium inflatum）为外群分别构建了NJ和UPGMA系统树，结果显示洞庭湖和鄱阳湖为一族群，太湖、巢湖和洪泽湖为一族群。     

西北太平洋鸢乌贼种群遗传结构

为检测西北太平洋鸢乌贼种群遗传结构,采用线粒体DNA细胞色素b基因(Cytb)序列分析方法对鸢乌贼东海群体、南海群体与菲律宾海群体进行遗传变异分析。结果显示,(1)所有群体总的单倍型多样度与核苷酸多样度分别为0.982±0.006、0.012±0.006;菲律宾海群体对应的遗传多样度均最高,分别为0.973±0.014、0.015±0.008;南海群体与东海群体的单倍型多样度分别为0.959±0.026、0.943±0.031,核苷酸多样度均为0.006±0.003。3个地理群体均具有较高的遗传多样性水平。(2)分子方差分析结果显示,34.6%的遗传变异来自于群体间,群体间遗传分化极显著。两两群体间Fst分析表明,西北太平洋鸢乌贼群体间均具有极显著的遗传分化。构建的单倍型邻接系统树和最小跨度树显示,西北太平洋鸢乌贼群体存在明显的系统发育谱系结构(谱系A、B、C),3个谱系单倍型类群间也存在极显著的遗传分化(Fst=0.735~0.805)。(3)中性检验和核苷酸不配对分析结果均表明,谱系B可能经历过近期群体扩张事件,发生群体扩张的时间在10.3~12.5万年前。综合分析认为,西北太平洋鸢乌贼的种群遗传结构模式及系统发育地理格局模式是由其栖息地海洋环境与更新世气候变化共同塑造的。建议在渔业管理上将3个地理群体划分为3个独立的管理单元。     

Characterization of Glycerophospholipid Molecular Species in Two Species of Arcidaes (Scapharca subcrenata and Scapharca broughtonii)

In this study, the molecular species of glycerophosphocholine (GPCho), glycerophosphoethanolamine (GPEtn), glycerophosphoserine (GPSer), glycerophosphatidylinositol (GPIns), lysoglycerophosphocholine (LGPCho), and lysoglycerophosphoethanolamine (LGPEtn) from arcidaes Scapharca subcrenata and Scapharca broughtonii were characterized using direct infusion-tandem mass (shotgun) method for the first time. At least 201 and 303 glycerophospholipid (GP) molecular species were characterized, respectively. Most of the predominant GP molecular species contained polyunsaturated fatty acids (PUFA), mainly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Meanwhile, the arcidae lipids contained a high proportion of polar lipids (81.57–86.72% of total lipids). Among PL, phosphatidylcholine (30.53–32.04 mol%) and phosphatidylethanolamine (21.48–26.71 mol%) were dominant.     

Genetic variation of hatchery and wild stocks of the pearl oyster <Emphasis Type="Italic">Pinctada fucata martensii</Emphasis> (Dunker, 1872), assessed by mitochondrial DNA analysis

In order to provide baseline information for the genetic resources, genetic variation in wild and cultured Pinctada fucata martensii from southern Korea and Japan was studied using nucleotide sequence analysis of 379 base pairs (bp) in the mitochondrial cytochrome oxidase subunit I gene (COI). The study included three hatchery stocks from Korea (Tongyeong) and Japan (Mie and Tsushima) and one wild stock from Korea (Geoje). A total of 3 haplotypes were identified in hatchery stocks of 78 individuals, of which 63 individuals shared 1 haplotype. Overall, nucleotide diversity (π) was low, ranging from 0.000 to 0.002, and haplotype diversity (h) ranged from 0.000 to 0.541. Considerably low haplotype and nucleotide diversities in hatchery stock indicated that low effective population size and consecutive selective breeding of P. fucata martensii could be responsible for the reduction in genetic variation. The wild stock exhibited low haplotype diversity (0.507 ± 0.039) with two shared haplotypes. The results of the present study with first record of wild pearl oyster in Korea support the possibility that the transplanted pearl oyster for overwintering experiments could have survived in winter. In order to enhance and/or maintain genetic diversity in the hatchery stock, further research should be directed toward genetic monitoring and evaluation of the hatchery and wild pearl oysters.     

Phylogeography,genetic stocks,and conservation implications for an Australian endemic marine turtle

1. Identification of the geographic extent of population boundaries, the distribution of genetic lineages, and the amount of genetic exchange among breeding groups is needed for effective conservation of vulnerable marine migratory species. This is particularly true of the flatback turtle (Natator depressus), which only breeds in Australia but has extensive migrations that can include international waters.
2. This study investigated the phylogeography and genetic structure among 17 flatback turtle rookeries across their range by sequencing an 810 bp portion of the mitochondrial DNA in 889 samples and genotyping 10 microsatellite loci in 598 samples.
3. There was low phylogenetic divergence among haplotypes and evidence of recent population expansion, likely in the late Pleistocene. A predominant haplotype was found across all rookeries, but other haplotype groups were regionally specific.
4. In general, there was agreement in patterns of genetic differentiation in the mitochondrial DNA and microsatellite data, and in some pairwise comparisons a higher mutation rate of microsatellites provided stronger evidence of differentiation.
5. These results suggest natal philopatry operates in the choice of breeding locations for males as well as females.
6. Evidence of genetic connectivity among neighbouring rookeries led to the identification of seven genetic stocks. Geographic boundaries of rookeries used by genetic stocks varied widely (160–1,300 km), highlighting a need for field studies to better understand movement patterns.
7. Hierarchical analysis of molecular variance identified significant genetic differentiation based upon genetic stock, nesting phenology (summer vs. winter nesters), and a west–east discontinuity across Torres Strait. A pattern of isolation by distance was identified, which was most strongly observed in the microsatellite data.
8. In combination with tagging and telemetry studies, these results will allow better quantification of stock‐specific threats along migratory routes and in foraging habitats. Implications of climate change will be stock specific and may depend upon the extent of genetic connectivity between neighbouring stocks.