鳙基于10个微卫星标记的亲子鉴定分析

为开展鳙(Hypophthalmichthys nobilis)家系选育工作,本研究进行了基于微卫星标记的亲子鉴定研究。试验中筛选了10个扩增效率较高的微卫星标记,通过引物荧光修饰,引物结合毛细管电泳分型技术,对鳙48尾亲本及384尾子代进行了基因分型,并计算了等位基因频率和模拟分析和亲子鉴定等分析。结果发现,各位点的等位基因数介于4~13之间,其中9个位点均具有较高的多态性和杂合度(PIC>0.5,He>0.5),研究发现位点的多态性信息含量(PIC)与亲本对排除率(E-PP)存在显著正相关(p<0.01)。模拟分析结果显示,该10个标记预计可用于已知性别的50组亲本(100尾)或未知性别的50尾亲本的鉴定分析(鉴别成功率>95%)。亲子鉴定发现,对试验中2个交配组(每组12对亲本)的鉴别成功率分别为98.96%和100%;且父母本对子代的贡献率存在极显著差异(p<0.01)。通过累积位点的鉴定分析发现,当标记数为7个和9个时分别能满足试验中12组和24组亲本对应子代的鉴定分析(鉴别率>95%),模拟分析和亲子鉴定分析成功率趋势基本符合。本研究所开发的亲子鉴定技术可为鳙家系选育提供技术支持。     

长江鲢、鳙个体发育过程中的表型变化

采用多元分析方法,对采自长江老江河国家级四大家鱼原种场255尾1—5龄鲢、鳙的30项形态度量数据进行了分析。聚类分析显示,鲢可明显地聚成欧氏距离相对较远的1龄组和2—5龄组,鳙也可聚成欧氏距离较远的1、2龄组与3—5龄组。判别分析显示,144尾鲢个体对所聚类的两个组别的判别准确率高达98.0%,111尾鳙个体对所聚类的两个组别的判别准确率也达90.7%。这表明,在幼体向成体的发育过程中,鲢和鳙在形态上均存在着两个具有显著差异阶段。单因素方差分析显示,鲢两个组别之间有18个参数差异极显著,4个差异显著,其中10个显著或极显著地变小,12个显著或极显著地变大。鳙两个组别间有极显著和显著差异的参数分别有14个和3个,其中极显著或显著差异变小和变大的分别有12个和5个。这表明在鲢、鳙个体发育过程中,分别有73.3%和56.7%的特征参数具有生长异速现象。因此,在根据形态鉴别物种、分析种群差异、确立种质标准和作生长退算时,均应考虑所选用特征的生长异速现象。     

Age, Growth, and Gonadal Characteristics of Adult Bighead Carp, Hypophthalmichthys nobilis, in the Lower Missouri River

Bighead carp were introduced into Arkansas in 1973 to improve water clarity in production ponds. Bighead carp subsequently escaped aquaculture facilities in the early 1980's and dispersed into the Mississippi and Missouri rivers. The first documentation of bighead carp reproduction in the Mississippi River system was in 1989. The population has increased in the Missouri River as is evident in their increased proportion in the commercial harvest since 1990. The effect of this exotic planktivore on native ecosystems of the U.S. has not been examined. Basic biological data on bighead carp Hypophthalmichthys nobilis in the Missouri River are needed to predict potential ecological problems and provide a foundation for manipulative studies. The objectives of this study were to assess age, growth, and gonadal characteristics of bighead carp in the Missouri River. Adult bighead carp in our sample varied from age 3 to age 7 and length varied from 475 to 1050mm. There was a large variation in length at age, and overall bighead carp exhibited fast growth. For example, mean back-calculated length at age 3 was 556mm. The sample was dominated by bighead carp from the 1994 year class. There was no difference in gonad development (i.e., gonadal somatic index, egg diameter) between winter and spring samples. Length of male bighead carp and GSI were not significantly correlated; however, females exhibited a positive linear relationship between length and GSI. In each ovary, egg diameter frequencies exhibited a bimodal distribution, indicating protracted spawning. Mean fecundity was 226213, with a maximum fecundity of 769964. Bighead carp in the Missouri River have similar life history characteristics to Asian and European populations. They have become well established in the Missouri River and it is likely that dispersal and population density will increase.     

Mitochondrial Genome Sequencing and Development of Genetic Markers for the Detection of DNA of Invasive Bighead and Silver Carp (Hypophthalmichthys nobilis and H. molitrix) in Environmental Water Samples from the United States

Invasive Asian bighead and silver carp (Hypophthalmichthys nobilis and H. molitrix) pose a substantial threat to North American aquatic ecosystems. Recently, environmental DNA (eDNA), genetic material shed by organisms into their environment that can be detected by non-invasive sampling strategies and genetic assays, has gained recognition as a tool for tracking the invasion front of these species toward the Great Lakes. The goal of this study was to develop new species-specific conventional PCR (cPCR) and quantitative (qPCR) markers for detection of these species in North American surface waters. We first generated complete mitochondrial genome sequences from 33 bighead and 29 silver carp individuals collected throughout their introduced range. These sequences were aligned with those from other common and closely related fish species from the Illinois River watershed to identify and design new species-specific markers for the detection of bighead and silver carp DNA in environmental water samples. We then tested these genetic markers in the laboratory for species-specificity and sensitivity. Newly developed markers performed well in field trials, did not have any false positive detections, and many markers had much higher detection rates and sensitivity compared to the markers currently used in eDNA surveillance programs. We also explored the use of multiple genetic markers to determine whether it would improve detection rates, results of which showed that using multiple highly sensitive markers should maximize detection rates in environmental samples. The new markers developed in this study greatly expand the number of species-specific genetic markers available to track the invasion front of bighead and silver carp and will improve the resolution of these assays. Additionally, the use of the qPCR markers developed in this study may reduce sample processing time and cost of eDNA monitoring for these species.     

新型湘云金鳙遗传多样性的RAPD及微卫星分析

湘云金鳙是本实验室经过近十年的工作选育出来的,该种鱼通体金红色,肉质鲜嫩,生长速度快,是一种很有价值的养殖鱼种和实验材料.随机选取12尾湘云金鳙(RBC)和12尾来自湘江流域的普通鳙鱼BC(作为对照)进行RAPD和微卫星分析.在优化RAPD检测条件基础上,从230个随机引物中筛选出来的45个扩增较好且多态性强的引物对这...     

青草鲢鳙四种鱼同工酶的比较研究

采用聚丙烯酰氨垂直板状连续电泳方法,对青草鲢鳙６种组织、１０种同工酶和蛋白质进行了电泳研究,并结合作者以前做的工作,对所研究的共２１种同工酶和蛋白质在４种鱼中的组织分布,位点表达及活性作了分析和总结。结果表明：４种鱼同工酶的种内组织分布差异大于种间差异;ＡＤＨ、ＡＭＹ、ＣＡＴ、ＥＳＴ、ＭＥ、ＰＯＸ、ＳＤＨ存在不同程度的种间差异,可作为种类的遗传标记;４种鱼ＭＤＨ、ＧＯＴ、ＡＬＰ、ＡＯ、ＳＯＤ同工酶谱非常相似,只在活性上略有差异;ＬＤＨ、Ｇ６ＰＤＨ和ＣＯＸ的酶谱特征可作为青草和鲢鳙两个亚科的鉴别标记。     

长江中游四大家鱼仔鱼营养状况的初步研究

通过形态观察,食性分析及消化系统组织学检测,对长江中游四大家鱼仔鱼的营养状况进行了初步评价,仔鱼具有匀称,消瘦和很消瘦三种体型,空肠率较高,草鱼,鲢,青鱼和鳙分别为45．16％－72．13％,30％－40．01％,31．25％－67．65％和87．50％,依据肝胰脏的肠和组织学特征,可以将天然仔鱼的营养水平划分为很好,较好的饥饿三种,1998年采自九江江段的样本中,遭受饥饿的草鱼,鲢,青鱼和鳙分别为40％,48．15％,46．15％和60％。     

鳙鱼染色体的DAPI核型分析

利用腹腔注射秋水仙素制备肾细胞染色体方法和DAPI（4＇,6＇-diamidino-2-phenylindole）荧光染色的方法,对鳙鱼（Aristichthys,nobills）的染色体组型和染色质的分布进行了研究。结果表明,其二倍体数目为2n=48,核型为30M＋14SM＋2ST＋2T。DAPI荧光染色显示间期细胞核中荧光亮度较为一致,提示异染色质在间期细胞核中分布比较均一。而DAPI荧光染色在第1和第4染色体的短臂上较为明亮,其余染色体上的明亮区都分布在着丝粒区域,表明第1和第4染色体上的异染色质主要集中在染色体的短臂上,其余染色体的异染色质主要分布在着丝粒区域。     

五大连池鳙的生长分析

为了分析五大连池鳙(Aristichthys nobilis)的生长情况,于2013和2014年在五大连池三池及二池随机采集223尾鳙样本,采用鳞片鉴定其年龄,退算体长、体重。同时采用von Bertalanffy生长方程分析了五大连池鳙体长、体重的生长规律。结果显示,鳙渔获物的平均体长、体重分别为54.61 cm和3 280.53 g,由3+~8+共6个年龄组组成,以5+龄个体为主,占总数的50.67%。五大连池鳙体长(L,cm)、体重(W,g)之间关系式为W=0.0289 L2.901(n=223,R=0.974)。体长、体重von Bertalanffy生长方程分别为:Lt=97.619[1﹣e﹣0.140(ti﹣0.004)]和Wt=17113.260[1﹣e~(﹣0.140(ti﹣0.004))]2.901。鳙体长生长速度曲线无拐点,随着年龄的增加生长速度逐渐降低。体重的生长速度曲线具有拐点,拐点年龄为7.61龄,所对应的体长和体重分别为63.94 cm和5 012.82 g。采用生长特征指数?对比分析五大连池与其他水域鳙的生长,结果显示五大连池鳙的生长较为缓慢,栖息环境的水温、饵料生物密度是影响五大连池鳙生长的重要因素。有效提高鳙生长速度的关键措施在于降低放养密度。     

不同日照时数对鲤,鲢,鳙鱼苗生长和存活的影响

不同日照时数对鲤、鲢、鳙鱼苗生长和存活的影响王吉桥,赵德树,张景全（大连水产学院养殖系,１１６０２３）（大连市碧流河水库渔场,１１６４２８）ＥｆｆｅｃｔｏｆＰｈｏｌｏｐｅｒｉｏｄｏｎｔｈｅＧｒｏｗｔｈａｎｄＳｕｒｖｉｖａｌｏｆＳｉｌｖｅｒＣａｒｐ,Ｂ...     

Centromere Localization for Bighead Carp (Aristichthys nobilis) through Half-Tetrad Analysis in Diploid Gynogenetic Families

Gene-centromere (G-C) mapping provides insights into structural and behavioural properties of chromosomes. In this study, G-C mapping using microsatellite markers and meiogynogenetic (meiotic gynogenetic) families were performed in bighead carp (Aristichthys nobilis, 2N = 48), which belongs to Cyprinidae. A total of 218 microsatellites were selected across 24 linkage groups (LGs) of a recently well-defined genetic linkage map for bighead carp, with 151 being heterozygous in at least one of six dams in diploid meiogynogenetic families. After tests for Mendelian segregation in two diploid control families, 103 microsatellites were used for G-C distance calculation in 383 gynogens. The second division segregation frequency (y) was computed through half-tetrad analyses, and the values ranged from 0 to 0.97 (mean 0.40). High G-C recombination frequencies (over 0.667) were observed in 18 (17.5%) of the loci examined, which revealed a low level of chiasma interferences compared with other fishes studied previously. Distribution of G-C distances across LGs ranged from 0 cM to 48.5 cM (mean 20 cM) under the assumption of complete interference. All 24 centromeres were localized according to their closest-related microsatellites at 95% confident intervals. The average distance between centromeres and their closest-linked markers was 6.1 cM with 15 out of 24 LGs having a distance below 5 cM. Based on the centromere positions in this study, we proposed a formula of 24 m/sm+24 t/st chromosomes with 92 arms for bighead carp, which was mostly in accordance with a previously reported karyotype for bighead carp (24 m/sm+24 st). These results of centromere localization provide a basic framework and important resources for genetics and comparative genomics studies in bighead carp and its closely-related cyprinid species.     

云斑尖塘鳢胚胎和早期仔鱼的发育

对云斑尖塘鳢(Oxyeleotris marmoratus)胚胎和早期仔鱼的发育进行了观察,详细描述了各发育阶段的形态特征。在28℃定水温条件下,云斑尖塘鳢的胚胎发育历时80h 30min,可分为24个发育分期。在整个发育过程中,眼、耳囊、心脏、消化道、肾脏、鳔、胸鳍和尾鳍等得到了优先发育。     

粘皮鲻虎鱼胚胎及仔鱼的发育

粘皮鲻鰕虎鱼(Mugilogobius myxodermus)的卵产出时为球型,直径0.40~0.60mm,具黏性,卵颜色淡黄,半透明,油球多且数量不固定。受精5~10min后吸水膨胀成椭圆形。水温16~18℃的条件下,胚胎发育共需138h。初孵仔鱼体长2.30~3.10mm,体高0.35~0.40mm,总肌节29~30。出膜30min~24h内鳔充气,从孵出到各鳍分化完全约需要40d。     

Maxillary Apparatus in the Feeding of the Silver Carp Hypophthalmichthys molitrix (Cyprinidae)

Journal of Ichthyology - The paper describes in detail the structure of the connective tissue and of the bone components of the maxillary apparatus of the silver carp Hypophthalmichthys molitrix as...     

Embryonic and Larval Development and Early Behavior in Grass Carp,Ctenopharyngodon idella: Implications for Recruitment in Rivers

With recent findings of grass carp Ctenopharyngodon idella in tributaries of the Great Lakes, information on developmental rate and larval behavior is critical to efforts to assess the potential for establishment within the tributaries of that region. In laboratory experiments, grass carp were spawned and eggs and larvae reared at two temperature treatments, one “cold” and one “warm”, and tracked for developmental rate, egg size, and behavior. Developmental rate was quantified using Yi’s (1988) developmental stages and the cumulative thermal units method. Grass carp had a thermal minimum of 13.5°C for embryonic stages and 13.3°C for larval stages. Egg size was related to temperature and maternal size, with the largest eggs coming from the largest females, and eggs were generally larger in warmer treatments. Young grass carp larvae exhibited upward and downward swimming interspersed with long periods of lying on the bottom. Swimming capacity increased with ontogeny, and larvae were capable of horizontal swimming and position holding with gas bladder emergence. Developmental rates, behavior, and egg attributes can be used in combination with physical parameters of a river to assess the risk that grass carp are capable of reproduction and recruitment in rivers.     

Phylogeography and population genetics of introduced Silver Carp (<Emphasis Type="Italic">Hypophthalmichthys molitrix</Emphasis>) and Bighead Carp (<Emphasis Type="Italic">H. nobilis</Emphasis>) in North America

The success of a biological invasion and the ability to control an invader may partially depend on the genetic diversity of the invasive species and the amount of dispersal and gene flow occurring throughout its introduced range. Here, we used nuclear microsatellites to analyze genetic diversity and structure and whole mitogenomic sequences to analyze the phylogeography of Silver Carp (SC; Hypophthalmichthys molitrix) and Bighead Carp (BHC; H. nobilis) across their North American ranges. Our objectives were to assess: (1) the number of mitochondrial haplotypes that were introduced and how they are distributed in North America, which may provide insight into the history of the invasion, (2) how genetic diversity compares between the native Asian and introduced North American populations, (3) how genetic variation is structured across the North American ranges of SC and BHC as well as between the two species, and (4) whether patterns of genetic diversity and structure are likely to affect success of environmental DNA programs for monitoring these species. In both species, we found relatively few mitochondrial haplotypes, and most were present throughout the range. For both SC and BHC, unique haplotypes were found only in a portion of the species’ range, possibly indicating the location of additional, more recent introductions. In both species, genetic diversity was moderately lower in North American populations (i.e., 75–90% of that found in Asian populations), but genetic diversity still remained high. We found very little population genetic structure, consistent with a rapidly spreading invasive species, and did not find evidence of cryptic interspecific hybrids. The markers developed for eDNA monitoring will be effective for detecting the majority of individuals of these species in North America. The relatively high level of genetic variation and lack of population structure of SC and BHC in North America indicate that genetic diversity likely will not limit their persistence and that high connectivity will likely complicate efforts to control these invasive species.     

草鱼与鲢鱼肥胖基因克隆与序列分析

肥胖基因产物leptin是调节哺乳动物摄食、能量代谢等生命活动的重要细胞因子。 应用RT-PCR和RACE法获得了草鱼(Ctenopharyngodon idellus)和鲢鱼(Hypophthalmichthys molitrix) leptin基因的全长cDNA序列分别为1 096 bp和1 176 bp,编码173和172个氨基酸。氨基酸序列同源性分析表明,草鱼和鲢鱼的leptin序列与其它鲤科鱼类leptin的同源性较高,而与其他鱼类的leptin同源性很低,但所有鱼类的leptin均含有用于形成二硫键的高度保守的半胱氨酸。系统进化树分析显示,草鱼和鲢鱼leptin与其他鱼类leptin聚于一进化分支。应用PCR和Genome Walker方法,进一步获得了草鱼和鲢鱼leptin基因的内含子和5′侧翼区序列。结果表明,获得的草鱼和鲢鱼leptin基因长度分别为2 129 bp和2 192 bp,含有与其他脊椎动物leptin相似的基因结构（含三个外显子和两个内含子）。本研究为深入研究鱼类肥胖基因结构功能关系与鱼类抗肥胖品系定向遗传选育奠定了良好的基础。     

Erythrocyte measurements of diploid and triploid Ctenopharyngodon idella×Hypophthalmichthys nobilis hybrids

Diploid and triploid hybrids resulted when grass carp, Ctenopharyngodon idella , and big-head carp, Hypophthalmichthys nobilis , were experimentally hybridized. Erythrocyte size can be used to distinguish C. idella × H. nobilis hybrids of different ploidies.     

鲢鱼轻酶解肌球蛋白的cDNA克隆及结构解析

与栖息温度相关的鲢鱼两种轻酶解肌球蛋白重链(light meromyosin,LMM)同工型(低温型,sc-w;高温型,sc-s)的氨基酸序列解析结果表明:sc-w与sc-s在LMM的氨基酸序列上显示91.8%的同源性,但与已经报道的草鱼(Ctenopharyngodon idella)低温型(gc10)有96.9%的...