呼玛河3种茴鱼遗传分化及属内地位的微卫星分析

茴鱼属(Thymallus)是黑龙江水系特有的珍稀濒危冷水性鱼类,本研究利用6对微卫星引物,对采自黑龙江上游呼玛河同域分布的黑龙江茴鱼(T.grubii)、下游黑龙江茴鱼(T.tugarinae)及待定名种(T.sp.)3种茴鱼的遗传多样性进行了比较研究,以探讨其种群遗传结构、分化水平及分类学地位,为制定保护管理策略提...     

05下游黑龙江茴鱼种群遗传变异及地理分化的微卫星分析

下游黑龙江茴鱼(Thymallus tugarinae)为黑龙江水系中国境内茴鱼属(Thymallus)新纪录种,也是黑龙江水系特有的珍稀濒危冷水性鱼类.本研究利用7对微卫星引物,对乌苏里江(WS)、黑龙江上游的呼玛河(HM)和额木尔河(EM)3个下游黑龙江茴鱼地理种群的遗传多样性进行了比较研究,以探讨其种群遗传结构及地理分化水平,并为制定保护管理策略提供遗传学依据.结果显示,WS、HM和EM种群的平均等位基因数(Na)分别为18.1、14.4,和11.7,平均杂合度(H)分别为0.816 5、0.807 2和0.818 4,多态信息含量(PIC)分别为0.891 3、0.805 3和0.757 7,种群中的特有等位基因数分别为33、16和7.遗传多样性水平以WS最高,HM次之,EM最低,在地理上呈现出由黑龙江下游至上游逐渐下降的趋势.3个种群间等位基因及基因型分布不均衡(P＜0.01),WS与HM、EM的遗传距离(Ds)分别为0.609 3、0.635 7,遗传分化系数(Fst)分别为0.068 0和0.0827,HM与EM亲缘关系最近,Ds和Fst分别为0.221 2和0.035 6,3个种群间的遗传距离及分化程度与地理距离呈正相关.AMOVA分子方差分析显示,6.35%的遗传变异来自于种群间,93.65%的变异来自于群体内个体之间.研究表明,黑龙江水系下游黑龙江茴鱼种群具有很高的遗传多样性水平,3个地理种群间产生了一定的遗传分化,推测其独特的生活史特性及地理距离是导致种群间产生遗传分化的主要原因.综合以上研究,建议将乌苏里江种群和黑龙江上游种群(呼玛河、额木尔河)作为下游黑龙江茴鱼的2个独特进化单元,并制订相应的保护管理策略.     

利用线粒体D—loop基因序列分析黑龙江上游北极茴鱼群体遗传结构

分布在黑龙江上游呼玛河和额木尔河等水域的北极茴鱼一新纪录种Thymallus sp,种群数量少、分布范围狭小,群体资源面临濒危。本研究采用PCR技术分析了黑龙江上游的北极茴鱼线粒体DNA控制区的序列,共获得了1104bp核苷酸全序列,与分布于勒拿河的该种茴鱼群体进行比较。结果显示：在23尾个体中共检测到3个单倍型（Hap1、Hap2,及Hap3）,其中Hap3为黑龙江和勒拿河共享;利用单倍型构建的分子系统树和网络图显示,黑龙江群体可能起源于勒拿河。黑龙江上游群体的遗传多样性较低,单倍型多样性指数（0.530）和核苷酸多样性（0.00053）远低于同域分布的其它种茴鱼;黑龙江群体与勒拿河群体的Fst为38.56%（P〈0.05）,已产生一定的遗传分化。     

黑龙江上游3种茴鱼分类地位的线粒体D-loop序列变异分析

基于茴鱼属(Thymallus)鱼类的线粒体D-loop片段全序列变异,对分布于黑龙江上游中国境内的呼玛河、额木尔河等水系中形态特征差异显著且同域分布的3种茴鱼共24个个体进行分子系统发育关系研究,以探讨其在属内的分类学地位.结果显示,分布于黑龙江上游的茴鱼群体在属内显著分为3个独立进化分支,平均序列分歧为0.048～0.062,已达到种间分化水平.经形态学鉴定,其中2个分支分别为黑龙江茴鱼(T.grubii)和下游黑龙江茴鱼(T.tugarinae),而另外1个分支应为1个新种;且该种与分布于勒拿河上游的勒拿河茴鱼((T.sp.)构成单系群,平均序列分歧仅为0.006,即黑龙江上游和勒拿河上游的待定名茴鱼为同一种,结合文献,该茴鱼被确定为北极茴鱼(T.arcticus)的1个亚种.综合3种茴鱼在黑龙江水系的地理分布现状,对其起源及演化进行了初步推测.     

利用封闭式循环水系统培育北极茴鱼苗种试验

正北极茴鱼(Thymallus arcticus)隶属于鲑形目、鲑科、茴鱼亚科、茴鱼属,地方名花棒子、花翅子,是一种冷水性洄游鱼。主要分布于我国的新疆额尔齐斯河水系的别列孜克河、哈巴河、布尔津河及其上游的喀纳斯湖和库依尔特河等河流中,在黑龙江水系上游区域的额木尔河、呼玛河等支流也有分布。北极茴鱼主要栖息在水质清澈、中低层水域环境中,性凶猛,主食动物性活饵,成鱼个体较小,故种群生存能力较弱,又因人工捕捞和水利设施阻隔等因素,野生种群资源急剧减少。     

同域分布两种鲤科鱼类种群遗传结构的比较

为了解河南省境内黄河水系和卫河水系棒花鱼和（歺又鱼）条的鱼类遗传多样性现状，利用线粒体细胞色素b（cytb）基因，对这两种鱼类开展群体遗传学研究。结果显示，棒花鱼和（歺又鱼）条群体多态性较低；系统发育树和单倍型网络图结果显示棒花鱼和（歺又鱼）条均由3个线粒体谱系组成；棒花鱼和（歺又鱼）条种群间均存在明显的遗传分化，种群间中等偏低的基因流解释了两种鱼类均具有较低的遗传多样性和明显的种群分化；群体历史动态分析显示，棒花鱼和（歺又鱼）条种群数量分别在距今约0.019百万年和0.026百万年开始锐减。鉴于群体多态性，种群分化和群体历史动态研究，这两种鱼类的遗传多样性均较低，需加强对其资源保护。     

长江流域铜鱼和圆口铜鱼的遗传多样性

利用线粒体DNA控制区序列多态性分析了长江流域铜鱼（Coreius heterodon）和圆口铜鱼（Coreius guichenoti）各4个群体的遗传结构；同时利用9对自行开发的多态性微卫星标记分析圆口铜鱼4个群体的遗传结构。结果显示，铜鱼线粒体DNA（mtDNA）D-loop序列共检出22个多态位点，28种单倍型，平均单倍型多样性指数（h）和核苷酸多样性指数（7r）分别为0．849和0．00257。圆口铜鱼线粒体DNA（mtDNA）D-loop序列共检出18个多态位点，28种单倍型，平均单倍型多样性指数和核苷酸多样性指数分别为0．902和0．00424。分子变异分析（AMOVA）结果提示，铜鱼和圆口铜鱼分别有98．80％和99．17％的遗传变异发生于群体内部，表明铜鱼和圆口铜鱼未出现种群分化。选用的9个微卫星标记在圆口铜鱼群体中共检测到48个等位基因；群体平均观测杂合度在0．631～0．753之间；平均期望杂合度为0．598-0．728：平均多态信息含量为0．548～0．670。结果表明，长江流域铜鱼遗传多样性较低，长江上游圆口铜鱼遗传多样性较高，且均未出现种群遗传分化。圆口铜鱼SSR固定指数为0．12l58，高于D．1oop固定指数，显示SSR标记对圆口铜鱼群体间遗传差异的检测更为灵敏。[中国水产科学，2008，15（3）：377-385]     

极具养殖价值的名特优鱼类——黑龙江茴鱼

黑龙江茴鱼系黑龙江省特有的名贵冷水性食用鱼类,其个体较小,生长速度相对稍慢。近年来,随着人们消费观念的转变,肉质鲜美、营养丰富的黑龙江茴鱼逐渐成为广大消费者青睐的名优水产品。由于天然水域的产量非常有限,人工养殖具有广阔的市场前景。一、习性与特征黑龙江茴鱼属鲑形目、茴鱼科、茴鱼属,俗称斑鳟子等。在我国产于黑龙江上游、嫩江上游、牡丹     

基于线粒体Cytb基因序列的西江流域广西境内卷口鱼遗传多样性分析

为了解西江流域广西境内卷口鱼(Ptychidio jordani)种群遗传结构及分化程度,采用线粒体Cytb基因序列对西江流域广西境内6个江段的139尾野生卷口鱼的遗传多样性进行了分析。结果显示,线粒体Cytb基因长度为1 053 bp,碱基T、C、A、G的平均含量分别为29.1%、27.7%、29.3%、13.9%,其中A+T (58.4%)高于C+G(41.6%)。共定义20个单倍型,并聚为2个分支,未观察到明显的地理聚群。6个卷口鱼群体的平均单倍型多样性和平均核苷酸多样性分别为0.768 2、0.002 3,其中红水河群体(单倍型多样性h=0.748 7,核苷酸多样性π=0.003 3)遗传多样性最高,柳江群体(h=0.274 4,π=0.000 4)和左江群体(h=0.374 7,π=0.000 3)的遗传多样性相对较低。卷口鱼总体的遗传分化指数(F_(ST))为0.461 4 (P0.01),表现出较大的遗传分化。两两群体间遗传分化结果显示,左江和柳江种群之间的遗传分化程度最大,而柳江和西江之间最小。AMOVA分析表明西江流域的卷口鱼群体遗传变异一半来自群体内(53.86%),一半来自群体间(46.14%)。中性检验(Tajima's D=-1.082 8,P0.05;Fu's Fs=-6.572 5, 0.01P0.05)与碱基错配分布分析表明西江流域卷口鱼种群大约在0.07～0.187 Ma经历了种群扩张。综上,西江流域广西境内的卷口鱼柳江群体和左江群体遗传多样性较低,总群体分化程度较大,但仍属于一个种群,其中空间距离与地理阻隔对卷口鱼的遗传分化具有一定的促进作用。     

基于AFLP分子标记的杂色杜父鱼遗传多样性分析

为了解吉林省杂色杜父鱼野生群体遗传多样性，应用扩增片段长度多态性（amplified fragment length polymorphism, AFLP）标记技术，对漫江、三道松江河和鸭绿江上游采捕的3个杂色杜父鱼群体总计30个个体的遗传多样性、遗传变异及遗传相似性进行分析，并构建系统进化树。8组引物组合总计扩增出915个条带，其中多态性位点843个，占比92.1%。3个群体的观测等位基因数（N_a）分别为1.316 6、1.306 7、1.203 7,有效等位基因数（N_e）分别为1.151 7、1.134 2、1.111 9,Nei’s基因多样性指数（H）分别为0.091 2、0.081 6、0.067 0,Shannon’s信息指数（I）分别为0.141 0、0.127 3、0.102 0,其中漫江群体的遗传多样性水平最高，鸭绿江上游群体最低；3个群体的遗传分化系数(G_st)平均值为0.286 0,表明3个群体之间发生了一定程度的分化；30个个体的遗传相似性在0.809 0～0.945 6,表明所采集的30尾杂色杜父...     

Habitat characteristics affecting occurrence of a fluvial species in a watershed altered by a large reservoir

Flooding river valleys following construction of dams restrict fluvial environments to reaches that were formerly headwaters. Whether remaining habitat is suitable for all life stages of fluvial species is poorly understood. A fluvial species, Arctic grayling Thymallus arcticus, showed a dramatic decline following flooding of the Upper Peace River and the formation of the Williston Reservoir. We related landscape and field site‐specific features with occurrence of juvenile Arctic grayling using an information theoretic approach. For the landscape model, an association was identified between stream order and Arctic grayling occurrence although stream order alone was a poor predictor. A positive association between juvenile Arctic grayling occurrence and distance from the Williston reservoir and stream order, as well as a negative association with water temperature and temperature variance, was deemed important for the field site model. Both modelling approaches indicated size of stream system to be an important influence on occurrence of juvenile grayling in the Williston watershed. River length required for suitable river habitat for salmonids has not previously been identified, but should be factored into future management plans when evaluating the impact of proposed hydroelectric dams and subsequent flooding of river systems.     

Post-stocking dispersal of European grayling,Thymallus thymallus (L.), in a semi-natural experimental stream

Abstract Artificially hatched and reared juvenile European grayling, Thymallus thymallus (L.), from three natural populations were released into an experimental stream. Two of the populations were from lakes and the third population was from a river. Post-stocking dispersal was studied by recapturing fish in traps and by electric fishing after each of the stocking exercises. On all three occasions the first fish to be recaptured were caught in a trap about 400 m downstream from the release site, within 5–12 min of release. Proportions of stocked grayling juveniles recaptured by electric fishing in the fast-flowing section of the experimental stream (the rapids) were 9.9%, 46.7% and 16.6% after the first, second and third stocking, respectively. No significant population-specific difference was found between fish caught in the downstream trap and fish recaptured in the rapids. The uppermost part of the rapids (stretch 1) and the catch in the upstream trap was dominated by fish of riverine origin. Post-stocking migration was influenced by sex but not by body size. Females showed a greater propensity to stay in the rapids compared with males. Implications of the results for stocking grayling in natural streams and regulated rivers are discussed.     

呼玛河哲罗鱼遗传多样性的AFLP分析

2002年4月在黑龙江水系呼玛河塔河河段100 km范围内,用三刺网捕获索饵洄游哲罗鱼(Hucho taimenPallas)个体20尾,用12对引物对其进行AFLP分析,每对引物组合扩增的位点在39~53之间,共扩增出了565个位点,多态性位点为477个,多态性比例为84.43%,平均Nei氏基因多样性指数H为0.386 7,平均Shannon氏指数I为0.5102,个体间平均遗传距离为0.4552,UPGMA聚类结果显示呼玛河水系哲罗鱼群体内有严重的近亲交配现象。研究结果表明呼玛河水系哲罗鱼具有适度的遗传多样性,这对呼玛河水系哲罗鱼资源量的恢复有着至关重要的作用。     

Mass-marking of grayling, Thymallus thymallus (L.), larvae by fluorochrome tagging of otoliths

Abstract Larvae of grayling, Thymallus thymallus (L.) were immersed in tetracycline hydrochloride (TC) and alizarin red S (AL) solutions for 3–4 h at concentrations of 300 and 70 ppm, respectively. Fluorescent marks on the otoliths from the marked fish were identified using a UV-light microscope. Results obtained with AL were superior to TC in marking otoliths. Fish kept in an aquarium retained good marks for 842 days. Stocking of the Carpathian River Ochotnica with marked grayling larvae revealed good mark retention for 128 days and a survival rate of approximately 12.2%.     

Interspecific germ cell transplantation: a new light in the conservation of valuable Balkan trout genetic resources?

Interspecific transplantation of germ cells from the brown trout Salmo trutta m. fario and the European grayling Thymallus thymallus into rainbow trout Oncorhynchus mykiss recipients was carried out in order to improve current practices in conservation of genetic resources of endangered salmonid species in the Balkan Peninsula. Current conservation methods mainly include in situ efforts such as the maintenance of purebred individuals in isolated streams and restocking with purebred fingerlings; however, additional ex situ strategies such as surrogate production are needed. Steps required for transplantation such as isolation of high number of viable germ cells and fluorescent labeling of germ cells which are to be transplanted have been optimized. Isolated and labeled brown trout and grayling germ cells were intraperitoneally transplanted into 3 to 5 days post hatch rainbow trout larvae. Survival of the injected larvae was comparable to the controls. Sixty days after transplantation, fluorescently labeled donor cells were detected within the recipient gonads indicating successful incorporation of germ cells (brown trout spermatogonia and oogonia—27%; grayling spermatogonia—28%; grayling oogonia—23%). PCR amplification of donor mtDNA CR fragments within the recipient gonads additionally corroborated the success of incorporation. Overall, the transplantation method demonstrated in this study presents the first step and a possible onset of the application of the germ cell transplantation technology in conservation and revitalization of genetic resources of endangered and endemic species or populations of salmonid fish and thus give rise to new or improved management strategies for such species.     

Development of sperm vitrification protocols for two endangered salmonid species: the Adriatic grayling, <Emphasis Type="Italic">Thymallus thymallus</Emphasis>, and the marble trout, <Emphasis Type="Italic">Salmo marmoratus</Emphasis>

Vitrification was applied to the sperm of two endangered fish species of So?a River basin in Slovenia, the Adriatic grayling (Thymallus thymallus) and marble trout (Salmo marmoratus) following testing different cooling devices and vitrifying media. Sperm was collected, diluted in species-specific non-activating media containing cryoprotectants, and vitrified by plunging directly into liquid nitrogen without pre-cooling. Progressive motility, curvilinear velocity, and straightness of fresh and vitrified-warmed sperm were evaluated with computer-assisted sperm analysis (CASA). Fertilization trials were carried out to test the effectiveness of vitrification in the case of grayling. A protocol utilizing a glucose-based extender, 30% cryoprotectants (15% methanol + 15% propylene glycol), 1:1 dilution ratio, and droplets of 2 μl on a Cryotop as cooling device yielded the highest post-thaw motility values for both Adriatic grayling (7.5?±?6.5%) and marble trout (26.6?±?15.8%). Viable embryos were produced by fertilizing eggs with vitrified grayling sperm (hatching 13.1?±?11.7%, control hatching 73.9?±?10.4%). The vitrification protocol developed in this study can be utilized in the conservation efforts for the two species as an alternative to slow-rate freezing when working in field conditions or when specific equipment necessary for slow-rate freezing is not available.