基于线粒体Cytb基因全序列的松江鲈群体遗传结构分析

对松江鲈(Trachidermus fasciatus Heckel)中国沿海7个群体和日本有明海群体的线粒体Cytb基因全序列进行了测定、分析。结果显示: 47个个体共检测到31个单倍型, 8个群体均呈现出较高的单倍型多样性(0.60-1.00)和较低的核苷酸多样性(0.0005-0.0041)的特点。AMOVA分析结果及单倍型邻接关系树和单倍型网络关系图均显示松江鲈分为中国和日本两个世系, 而中国世系的7个群体未呈现出明显的地理遗传结构。基于核苷酸Kimura双参数替代模型计算得出的中国和日本两个世系的净遗传距离, 再参照其他硬骨鱼类线粒体Cytb基因2%/Ma(百万年)的分歧速率, 推测松江鲈中日两个世系间分化时间约为41万年前。对中国世系进行群体历史动态分析, 中性检验结果均为负值且显著, 核苷酸不配对分布呈单峰型, 表明松江鲈中国世系曾发生过群体扩张, 其扩张时间大约为12万年前。       

扬子鳄饲养种群线粒体DNA控制区的序列多态性

扬子鳄(Alligator sinensis)是中国特有的珍稀爬行动物,至2000年,野生扬子鳄的个体数已不足150条,作为保护这一物种的措施之一,先后于80年代初建起了2个养殖场,现人工繁殖的扬子鳄总数已达9000余条。为揭示扬子鳄种群遗传多样性,从两个饲养种群中采集了42个个体的样品,其中宣州样品33个(xZSP),长兴样品9个(CxSP),用PCR方法扩增mtDNA控制区,扩增产物纯化后直接用ABI310全自动遗传分析仪荧光标记测序,得到其中39个个体的血DNA控制区5’端462bP的序列。经比对发现,39个个体间的5’端mtDNA控制区没有任何变异位点,共享一种单元型,提示扬子鳄饲养种群的遗传多样性非常贫乏,造成这一结果的主要原因是近50年来,扬子鳄种群衰退和数量迅速减少导致的遗传多样性丢失,其次是人工繁殖的群体同时受到始创者数量较少产生的瓶颈效应影响。针对扬子鳄遗传多样性的现状,作者最后就这一濒危动物遗传多样性的保护对策提出3点建议。     

基于线粒体DNA的中国亚洲象种群遗传多样性及种群遗传结构

亚洲象是我国国家一级保护动物.本文利用非损伤性取样法,以亚洲象粪便中脱落的肠道上皮细胞为DNA来源,选用线粒体DNA作为分子标记,对分布于我国境内的亚洲象种群的遗传结构和种群遗传多样性进行研究.本研究得到mtDNA序列片段长度为556 bp,经对178个个体进行扩增结果分析,共得到24个单倍型.在5个地理种群中,除南滚河种群外,其他4个种群中的114个个体共享同一单倍型,南滚河种群与其他种群间未观察到共享单倍型.系统发生分析,观察到中国境内现有亚洲象种群在进化上分为两大分支,α和β.其中分支α中包含除南滚河种群外的4个地理种群,分支β仅含有南滚河种群,表明南滚河种群与其他4个地理种群间存在明显分化.遗传多样性分析结果表明,中国境内的亚洲象种群的遗传多样性水平较低,分析原因认为是栖息地破碎化阻断了种群间有效的基因交流.     

基于线粒体控制区的序列变异分析中国东南部沿海凤鲚种群遗传结构

为了解中国东南部沿海凤鲚(Coilia mystus)的种群遗传多样性和遗传结构, 本文分析了长江口(CJ)、钱塘江口(QT)、闽江口(MJ)和九龙江口(JL)4个凤鲚地理群体的mtDNA控制区561 bp片段的序列变异。65尾样本共检测到28个单元型。4个群体总的单元型多样性和核苷酸多样性均较高(h = 0.9433 ± 0.0168, π = 0.0317 ± 0.0158), 但单个群体的核苷酸多样性水平却很低, 其中以CJ最高(π = 0.0080 ± 0.0046), MJ最低(π = 0.0015 ± 0.0013)。MJ与JL群体之间以及CJ与QT群体之间的平均K2P遗传距离很小, 分别为0.3%和0.8%; 而CJ、QT分别与MJ、JL群体之间的遗传距离均较大, 达到了6%。采用最大似然法(ML)、最大简约法(MP)和邻接法(NJ)分别构建的单元型间的系统发育树揭示, 4个凤鲚群体构成CJ-QT 和MJ-JL 2个支系, 且具有极高的支持率。单元型的网络分析也显示这两个支系间有高达28步的突变次数。AMOVA分析显示大部分的遗传变异来自这两支系群体间(90.77%), 表明凤鲚群体间存在着显著的地理分化。种群分化指数和基因流分析也表明, 支系间群体有着明显的遗传分化(F_ST > 0.9, N_m < 0.03)。所有分析结果支持所研究的凤鲚标本属于两个不同的地理种群, 且种群的分化至少已达到亚种水平。采用BEAST和TRACER软件得到凤鲚两个亚种的最近共同祖先约在0.34–0.46百万年前, 处于更新世晚期, 推测可能是第四纪晚期的气候旋回和海平面的升降导致了凤鲚的种群分化。     

基于线粒体COI基因分析钩手水母的群体遗传结构

钩手水母(Gonionemus vertens)为大西洋和太平洋广布种, 是我国习见的有毒水母种类之一。本文对采自黄渤海海域4个地理群体的104个钩手水母线粒体COI基因序列进行扩增, 并结合GenBank上其他182个钩手水母同源序列进行序列变异分析。在286个基因序列中共检测出52个多态位点, 定义了14种单倍型。总群体的单倍型多样性和核苷酸多样性分别为0.743 ± 0.012和1.046% ± 0.097%, 与其他几种大型水母相比, 钩手水母总群体的遗传多样性处于较高水平。AMOVA结果显示, 60.17%的分子变异源于群组间, 13.37%的分子变异源于群体内, 26.46%的分子变异源于组内群体间, 群组间、群体内和组内群体间的遗传分化均极显著。F_st值统计检验表明, 中国厦门群体与乐亭、东营、烟台、大连群体间存在显著的遗传分化, 大连与东营、烟台群体间也存在显著的遗传分化。系统分析结果显示, 钩手水母群体间存在2个明显的单倍型谱系分支。不同的钩手水母地理群体间具有复杂的遗传模式, 钩手水母复杂的生活史、扩散能力、地理隔离和海流分布可能是影响钩手水母遗传结构的重要因素。     

基于线粒体DNA D-loop区部分序列分析舟山海域带鱼种群遗传结构

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草鱼中国土著群体与欧美日移居群体遗传差异的线粒体序列分析

草鱼是中国的土著鱼类,自20世纪60年代以来已被移居到100多个国家和地区,主要用来控制水草或水产养殖。本研究通过线粒体D-Loop区（764bp)和COII＋tRNA基因(719bp)序列分析,了解草鱼的中国土著群体（长江、珠江、黑龙江）和国外移居群体（匈牙利多瑙河、美国密西西比河、日本利根川河）之间以及各地方群体间的遗传差异。结果表明,中国土著草鱼群体的遗传变异高于国外移居群体;分子方差分析（AMOVA）表明,不同草鱼群体的遗传变异主要来自群体内,而不同地域间的差异极少;群体两两间Fst值比较表明,大多数群体之间遗传差异极显著;由TCS构建的单倍型网络结构图显示,长江草鱼群体是最原始的群体,其他水系草鱼均由长江群体演化而来;通过基因流分析发现,匈牙利多瑙河群体和日本利根川河群体来自长江和黑龙江群体,美国密西西比河群体除引自长江、黑龙江水系外,还有部分引自于珠江水系。     

基于ITS序列对栽培中国樱桃遗传多样性及其群体遗传结构的分析

该研究基于ITS序列对栽培中国樱桃[Cerasus pseudocerasus(Lindl.)G.Don]18个群体共154个个体的遗传多样性及其群体遗传结构进行了分析,以期从DNA序列水平上揭示栽培中国樱桃种质资源的遗传背景,为保护和利用中国樱桃种质资源提供理论依据。结果表明:(1)154条ITS序列比对后共定义了11个单倍型,表现出较低的遗传多样性(h=0.559 0,π=0.001 2),群体间遗传多样性也表现出较大差异(h=0~0.905 0,π=0~0.006 1)。(2)群体分析显示,群体间遗传分化水平较低(FST=0.140 0),只有14%的遗传变异来自群体间,而86%的遗传变异来自群体内部。研究认为,栽培中国樱桃在驯化过程中所产生的奠基者效应以及瓶颈效应可能是导致群体遗传多样性丢失的主要原因,而较长的世代周期及较短的分化时间可能导致了群体间低的遗传分化;因此,对栽培中国樱桃种质应采取就地保护策略;若需迁地保护种质建议减少采样群体数而增加群体内个体数量的采样策略。     

我国沿海缢蛏群体遗传结构的mtDNA-COⅠ分析

采集了我国沿海共计9个缢蛏(Sinonovacula constricta)地理群体的197个样本,分别是北部组群的3个群体:辽宁省庄河群体(ZH),天津市汉沽群体(HG),山东省海阳群体(HY);中部组群的3个群体:江苏省盐城群体(YC),上海市崇明县东滩群体(DT)和堡镇群体(BZ);以及南部组群的3个群体:浙江省宁波群体(NB),浙江省台州群体(TZ)以及福建省宁德群体(ND)。利用线粒体COⅠ标记分析了9个群体的遗传多样性和遗传分化。结果表明,在共计197个个体中检测到125个单倍型和96个变异位点,核苷酸多样性指数位于2.1764～7.4970之间,其中中部组群的群体遗传多样性指数最高。AMOVA分析结果显示,组间遗传变异量占总变异的80.27%,18.74%来自于群体内,只有0.99%来自于组内群体间。群体间遗传分化系数位于0.0219～0.8706之间,不同群体间具有一定的遗传分化,尤其是中部群体与其他群体间遗传分化值达到了0.8以上,为极高度分化。遗传距离和聚类结果显示,北部3群体和南部3群体首先聚在一起,之后与中部3群体聚类。     

中国大陆梅花鹿mtDNA控制区序列变异及种群遗传结构分析

测定了37只中国大陆梅花鹿(Cervus nippon)不同种群mtDNA控制区5′端351 bp的序列，共发现23个变异位点，定义了5种单元型。分子变异分析表明，中国大陆梅花鹿出现了显著的种群分化(Φm＝0．45，Fst＝0．60，P＜0．001)，支持把分布于东北、华南和四川的梅花鹿种群归入各自独立的管理单元。中国大陆、日本南部和日本北部之间无共享单元型，且有25个鉴别位点。最小跨度网络图(Minimum spannlng network，MSN)和基于最大似然法和邻接法的系统发生分析均把单元型聚类为对应于中国大陆、日本南部和日本北部的三个单系，其中中国大陆和日本南部梅花鹿有相对较近的亲缘关系，支持日本梅花鹿的祖先通过至少两个大陆桥从亚洲迁移到日本的观点。     

Isolation of microsatellite DNA and analysis on genetic diversity of endangered fish,Hucho taimen (Pallas)

Hucho taimen (Pallas) is an endangered species in China. To evaluate the species’ genetic diversity and population genetic structure, we isolated six polymorphic microsatellite markers from its genomic libraries by (ACA)₁₆ enriched, and characterized using 61 wild individuals. The number of alleles per locus ranged from two to seven, expected heterozygosity ranged from 0.3200 to 0.7410, polymorphism information content ranged from 0.3047 to 0.6896. These markers will be useful for the genetic variation assessment of taimen.     

乌苏里江哲罗鲑的年龄结构、性比和生长

We report the current status of Taimen (Hucho taimen ) population in Wusuli River of China. The amount of catch per year was only 1 800 - 5 400 individuals ranging from 2 - 15 years old during 1998 - 2002. Of all the captured, individuals with sexual maturity were up to 30% - 70% (female maturity at 5 years old and male 6 years old). Sex ratio varied greatly among different localities. Its breeding population mainly consisted of males aged 5 - 10 years and females aged 8 - 13 years‘‘old, which spawned in May each year. Females breed once with about 4 000 - 23 000 eggs produced every 2 - 3 years, Taimen grows fast and average growth rate is about 10 cm per year prior to 10 years‘‘s old, which corresponded with von Bertelanffy‘‘s equation calculated as Lt = 246.41 [ 1 -e 0.0407(t 0.4625)] and Wt = 174 075.72 [ 1 -e ^0.0407(t-0.4622)12.9537, and the relationship between body length and weight was W 0.015018L^u2020.     

Analysis of genetic diversity in the endangered Hucho taimen from China

Hucho taimen are listed as endangered in China. The population size has declined recently, prompting an increase in the level of listing from grade three in 2002 to grade five in 2006. We analyzed the genetic diversity of wild populations using 17 microsatellite markers to establish a scientific basis for conservation of this species. We collected tissue samples from four populations in the Heilongjiang River basin: Huma River (HM), Hutou (HT), Haiqing (HQ), and Zhuaji (ZJ). A total of 21 loci were amplified, 18 of which were polymorphic. The number of alleles per locus ranged from 2 to 9 (mean: 4.1905). There were 13 highly polymorphic loci and 5 moderately polymorphic loci. Analysis of five genetic diversity parameters (Na, Ne, Ho, He, and PIC) suggested moderate levels of diversity within the populations. The populations were ranked HT > HQ > ZJ > HM, but the differences in diversity were not statistically significant (P > 0.05). A comparison of variation among all four populations suggested Hardy–Weinberg disequilibrium at 20% of the loci. Genetic differentiation (Fst) was 0.0644 and the gene flow among populations was estimated at 3.36 individuals per generation. The majority of diversity (93.88%) occurred among individuals within a population. In contrast, relatively little (6.12%) of the genetic diversity was distributed between the populations. An analysis of genetic differentiation and genetic distance between pairs of populations revealed that both parameters were higher in comparisons of the HM population to the HT, HQ, and ZJ populations than among the three latter populations. This suggests that the HM population has a distinct genetic structure. We hypothesize that habitat degradation and excessive fishing, not low genetic diversity, has caused the decline in H. taimen populations. However, this species should be protected from further declines in genetic diversity.     

Comparative mtDNA sequence (control region, ATPase 6 and NADH‐1) divergence in Hucho taimen(Pallas) across four Siberian river basins

Hucho taimen from eight populations spanning four drainage basins (Amur, Lena, Enisei and Khatanga) were analysed for nucleotide sequence variation across three mitochondrial genes (ATP6, NADH‐1 and control region). Samples of H. hucho , Brachymystax lenok (sharp‐snouted and blunt‐snouted forms) and Parahucho perryi were also included for comparison. Nucleotide variation across a total of 1826 base pairs in H. taimen revealed shared haplotypes between the Amur and Lena basins, further supporting a previous hypothesis of late to post‐Pleistocene hydrological exchange between these now disjunct basins. In contrast to an earlier study using the control region alone, clear phylogeographic structure was seen at a large geographic scale, reflected by two phylogroups, one corresponding to the Amur and Lena basins, and the other to the Enisei and Khatanga basins. Comparative rates of divergence revealed considerably faster and less heterogeneous substitution rates for the two coding genes, especially at interspecific levels compared to the mtDNA control region.     

不同月龄哲罗鲑主要形态性状与体重的关系

为确定影响哲罗鲑(Hucho taimen)体重的主要形态性状,随机选取5个生长阶段共计710尾哲罗鲑为实验对象(3月龄240尾、6月龄180尾、12月龄110尾、18月龄90尾、36月龄90尾),测量其体重(y)、体长(x_1)、体宽(x_2)、体高(x_3)、头长(x_4)、眼径(x_5)、尾鳍长(x_6)、尾鳍高(x_7)、尾柄高(x_8)9个性状。通过相关分析、通径分析和回归分析,计算相关系数、通径系数和决定系数,获得相关回归方程,对不同月龄间的形态性状对体重的影响程度进行了比较分析。结果表明,3月龄、6月龄、18月龄和36月龄个体体长对体重的直接作用较大,12月龄个体则是体宽对体重的影响较大。利用逐步回归分析方法,以体重为依变量(y),形态性状为自变量(x_1~x_8),建立了哲罗鲑不同月龄最优多元线性回归方程分别为:y3=﹣14.911+1.561 x_1+3.448 x_2+5.232 x_3;y6=﹣70.321+5.192 x_1+0.962 x_2+0.473 x_3;y12=﹣217.446+7.129 x_1+23.147 x_2+44.055 x_3;y18=﹣891.041+20.234 x_1+65.786x_2+53.266 x_3;y36=﹣5748.099+88.187 x_1+301.984 x_2+66.702 x_7。该研究结果为哲罗鲑选择育种的具体应用提供了重要参考依据。     

哲罗鱼消化系统器官发生发育的组织学观察

采用形态观察与连续组织切片技术,对哲罗鱼(Hucho taimen)胚胎期(水温7~8℃)和胚后期(水温3~14℃)消化系统的发生发育进行观察和研究。结果表明,哲罗鱼受精11d形成原始的消化管。受精18d肝原基出现,U型胃雏型形成。受精30d鱼体破膜,口能自由闭合。破膜8d后,齿形成,肛门与外界相通,消化道初步形成口咽腔、食道、U型胃、肠和肛门。破膜16d,胰及瓣囊出现,仔鱼消化系统初步具备了摄食和消化外源性食物的能力。破膜24d后,鱼体开始上浮,主动摄食,由内源性营养转向混合性营养。破膜30d后,卵黄囊完全被吸收,各消化器官功能和结构逐步发育完善,鱼体由混合性营养进入外源性营养阶段。此后随着鱼体的生长消化器官逐步发育成熟,舌齿和下颌齿分别为双排,胃腺发达,形成网状结构,幽门盲囊较多,肠为直行,肝和胰为相互分开的独立器官。     

Spawning migration and redd construction of Sakhalin taimen, Hucho perryi (Salmonidae) on northern Hokkaido Island, Japan

Spatial and temporal patterns of redd construction by Sakhalin taimen, Hucho perryi , were studied in small streams on northern Hokkaido, Japan. Spawning took place in late April with close relation to discharge patterns. Observed redd size (mean ± 95% C.I.) was 145 ± 11·8 cm in length and 84 ± 5·8 cm in width. The number of egg pockets per redd was either one or two, which is fewer than that of other salmonids reported in the literature. Spawning ground distributions in 1991 and 1992 were similar to each other in terms of the distribution mean and dispersion of the redds. Unlike other salmonids Sakhalin taimen probably constructed multiple redds during one spawning season.     

汉江上游太白河川陕哲罗鲑(Hucho bleekery Kimura)再发现

川陕哲罗鲑(Hucho bleekeri Kimura)是我国长江流域特有鱼类,近几十年来资源量急剧下降,陕西和四川等地历史分布区已多年未见其踪迹,濒临灭绝。2013年在汉江上游陕西省太白县太白河发现了19尾成体样本。本次发现是该物种在太白河消失15年后的再次发现,也是近30多年来该物种在分布区范围内同时被发现成体数量最大的一次。本次发现对川陕哲罗鲑资源保护和恢复有重要意义。     

Genetic structure and historical demography of Collichthys lucidus inferred from mtDNA sequence analysis

Fluctuations of global climate changes during the Pleistocene had a huge impact on the marginal seas of the Northwestern Pacific. To examine historical demography and the population genetic structure of Collichthys lucidus, a total of 151 individuals from 7 locations were sequenced at the 5′ end of mitochondrial DNA control region and 380-bp fragments were obtained. The results of AMOVA showed that the genetic variation among the two groups was 78.39 % (P?=?0.78) and among populations within groups was 0.43 % (P?=?0.02). Both mismatch distribution analysis and neutrality tests showed C. lucidus has experienced a recent population expansion. Three distinct clades suggested that C. lucidus might be isolated in the three marginal seas during the Pleistocene periods. No genetic divergence within groups should be explained by insufficient time to attain migration-drift equilibrium. The changes in sea level during the Pleistocene had a major effect on the C. lucidus phylogeographical pattern because it limited its dispersal and at times created isolated populations.     

The structure and dimensions of redds and egg pockets of the endangered salmonid, Sakhalin taimen

Each redd of the Sakhalin taimen Hucho perryi had a V–shaped pot that is specific to this species. The shape of each multiple–egg–pocket redd was a combination of one–egg–pocket redds, and the number of egg pockets in the redd could be estimated from the shape. False redds were small and did not have any V–shaped pots. The factors affecting redd size were examined numerically; consequently, numbers of egg pockets and eggs and female fork length were correlated significantly with tail length. Each egg pocket contained 11–920 eggs (mean: 546·7±95% CL 81·9) and the egg pocket depth was correlated significantly with female fork length. Mean number of redds per female was three, and the number of females could be estimated as one third of the redds.