基于mtDNA 16S rRNA序列的脉红螺(Rapana venosa)与红螺(R.bezoar)的分类学研究

采用线粒体DNA16SrRNA基因序列测定方法,对中国沿海3目8科的14种腹足类的系统演化关系进行了比较分析。结果显示,共测定了102个个体,获得线粒体DNA16SrRNA基因片段507bp的同源碱基序列;用最大简约法检测到保守位点87个,可变位点409个,简约信息位点254个;计算了碱基替换/颠换率的距离(R)和基于碱基替换+颠换率的距离(D),红螺属内的脉红螺和红螺R值为0.9367,大于0.5,属于种间标志值;采用Kimura2-parametermodel构建了邻接法(NJ)、最小进化法(ME)和最大似然法(MP)等3种系统树,脉红螺和红螺各自均为单系群(支持率=92—100),同时表明它们在检测的14种腹足类中属于相当进化种类。分析结果从线粒体基因序列的层次支持脉红螺与红螺属于不同种类的现代分类方法。     

金乌贼(Sepia esculenta)精子超微结构和低温损伤电镜观察

利用扫描和透射电镜技术研究了金乌贼成熟精子的超微结构,以及超低温冷冻对精子超微结构和形态的影响。结果表明,金乌贼精子为鞭毛型精子,外部附有光滑的裙边状被膜,分为头部、中部和尾部三部分。头部由顶体和细胞核组成,顶体位于细胞核前端,呈"囊"状。细胞核为长纺锤状。中部是由线粒体和垂直的中心粒构成,线粒体大小不一,多层分布。尾部细长,轴丝为典型的"9+2"结构。经超低温冷冻的精子被膜上出现很多凹陷;头部顶体破裂,大量的内容物流出,细胞核膜分离;线粒体解离,嵴变形;尾部缠绕在一起,少数鞭毛出现脱落。     

栉江珧第1卵裂抑制型雌核发育二倍体人工诱导的研究

利用6-二甲基氨基嘌呤(60 mg/L;6-DMAP)抑制第一卵裂,成功诱导出栉江珧(Atrina pectinata)雌核发育二倍体.研究结果表明在培养温度为24℃的条件下,受精后60 min用质量浓度为60 mg/L的6-DMAP处理栉江珧受精卵15 min进行雌核发育二倍体的诱导效果理想,D形幼虫发生率和诱导率分别为14.7%和22.7%.细胞学观察显示,6-DMAP阻止了纺锤体的形成和染色体的移动,导致一个融合的二倍性雌性原核的形成.本研究结果首次提供了栉江珧雌核发育二倍体的细胞学证据.     

中国沿海长蛸(Octopus variabilis)自然群体线粒体COI基因遗传多样性研究

对我国沿海5个自然群体长蛸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658bp核苷酸片段,其中碱基T、C、A和G的平均含量分别为37.02%、18.46%、30.15%和14.35%,AT的含量明显高于GC的含量。5个自然群体的长蛸中共发现18个变异位点,得到15个单倍型,包括4个共享单倍型,其中青岛群体的核苷酸差异数K以及平均核苷酸多样性指数Pi最高,烟台群体最低;群体间,青岛群体在群体间核苷酸差异数K、群体间平均每位点核苷酸替代数Dxy和群体间每位点净核苷酸替代数Da三个指标上都表现出比其它群体之间较高的水平,说明青岛群体具有最为丰富的遗传多样性。MEGA3.1软件计算5个群体间的Kimura2-paramter遗传距离,大连群体和青岛群体之间的遗传距离最远为0.0077,而大连群体与烟台群体之间的遗传距离最低,为0.0029。AMOVA分析表明,五群体间总遗传分化系数Fst=0.17410(P0.05),群体间遗传分化远小于群体内。NJ法和UPGMA法构建的分子进化树,5个地理群体的长蛸聚为两个族群,大连、烟台群体聚为一族群,青岛、连云港和舟山群体聚为另一族群。     

密鳞牡蛎的人工繁育

研究密鳞牡蛎(Ostrea denselamellosa)的亲体培育和苗种培育技术;研究盐度、温度、密度和饵料等因子对浮游幼虫生长及存活的影响,首次建立密鳞牡蛎的人工繁育技术。结果表明,排出体外的D形幼体在水温23~25℃,经约28 d出现眼点,进入附着变态阶段;幼虫培育的最佳盐度为30;在24~32℃,幼虫生长速度与温度呈正比,成活率与温度成反比;在培育密度为1,2,3和4/mL 4组中,1/mL组生长最快,但各组间存活率无显著差异;湛江等鞭金藻,50%湛江等鞭金藻+50%扁藻,50%湛江等鞭金藻+50%小球藻和小球藻4种饵料投喂组中,50%湛江等鞭金藻+50%扁藻组的生长显著高于其他组;不同附着基的附苗率依次为:扇贝壳>波纹板>筛绢>网片>聚乙烯薄膜。     

中国经济头足类增养殖现状及展望

头足类(Cephalopods)是软体动物门的重要组成部分,在中国海洋渔业中占有重要的经济地位。开展头足类增养殖,对于保护中国种质资源、提高头足类产量具有重要意义。本文就目前中国主要的经济头足类,包括金乌贼(Sepia esculenta)、拟目乌贼(S.lycidas)、虎斑乌贼(S.pharaonis)、无针乌贼(Sepiella japonica)、长蛸(Octopus minor)、真蛸(O.vulgaris)以及短蛸(Amphioctopus fangsiao)的繁殖习性、幼体培育、成体养成等各阶段生活史的研究进展和存在问题进行阐述,并对其发展前景进行展望,旨在为中国头足类养殖产业提供借鉴。     

Inheritance mode of microsatellite loci and their use for kinship analysis in the Pacific oyster （Crassostrea gigas）

Five full-sib families of the Pacific oyster(Crassostrea gigas) larvae were used to study the mode of inheritance at eight microsatellite loci,and the feasibility of these markers for kinship estimate was also examined.All eight microsatellite loci were compatible with Mendelian inheritance.Neither evidence of sex-linked barriers to transmission nor evidence of major barriers to fertilization between gametes from the parents was shown.Three of the eight loci showed the presence of null alleles in four families,demonstrating the need to conduct comprehensive species-specific inheritance studies for microsatellite loci used in population genetic studies.Although the null allele heterozygotes were considered as homozygotes in the calculation of genetic distance,offspring from five full-sib families were unambiguously discriminated in the neighbor-joining dendrogram.This result indicates that the microsatellite markers may be capable of discriminating between related and unrelated oyster larvae in the absence of pedigree information,and is applicable to the investigation of the effective number of parents contributing to the hatchery population of the Pacific oyster.     

Inheritance mode of microsatellite loci and their use for kinship analysis in the Pacific oyster （Crassostrea gigas）

Five full-sib families of the Pacific oyster （Crassostrea gigas） larvae were used to study the mode of inheritance at eight microsatellite loci, and the feasibility of these markers for kinship estimate was also examined. All eight microsatellite loci were compatible with Mendelian inheritance. Neither evidence of sex-linked barriers to transmission nor evidence of major barriers to fertilization between gametes from the parents was shown. Three of the eight loci showed the presence of null alleles in four families, demonstrating the need to conduct comprehensive species-specific inheritance studies for microsatellite loci used in population genetic studies. Although the null allele heterozygotes were considered as homozygotes in the calculation of genetic distance, offspring from five full-sib families were unambiguously discriminated in the neighbor-joining dendrogram. This result indicates that the microsatellite markers may be capable of discriminating between related and unrelated oyster larvae in the absence of pedigree information, and is applicable to the investigation of the effective number of parents contributing to the hatchery population of the Pacific oyster.     

不同浓度氨氮胁迫下长蛸(Octopus minor)的半致死浓度和急性毒性评估

作为重要的水质参数,高浓度的氨氮对水生动物产生有害影响。氨氮的安全浓度对海水养殖物种至关重要。本文采用静水法,在盐度30.0±1.0, pH 7.8±0.2,温度20.8±0.5°C条件下,以荣成月湖海区的长蛸(Octopus minor)成体为研究对象,分析了水体氨氮对其急性毒性作用。通过预实验获得长蛸成体可耐受的浓度范围,从中选取浓度0(对照)、87.0、124.5、143.5、180.3、204.0、242.5mg/L等7个浓度梯度测定长蛸成体耐受性,采用概率单位法、寇氏法和线性回归法分别计算48h、72h和96h的半致死浓度(LC50)。实验结果显示,氨氮毒性效应随着浓度和胁迫时间的增长而增加,氨氮胁迫48h、72h和96h的半致死浓度分别为187.93、154.61和124.92mg/L (χ2检验P0.05);显示氨氮对长蛸成体具有显著毒性作用。与其他水生动物相比,长蛸对于氨氮的耐受性较强,对氨氮毒性的自我调节能力较强,具体的调节机制有待进一步研究。     

魁蚶雌核发育的人工诱导

研究了魁蚶（Scapharca broughtonii）雌核发育的人工诱导条件。用强度为2561uw／（cm2．s）的紫外线（254nm）照射不同时间的精子与正常卵子受精,发现随照射时间的增加,卵裂率、早期胚胎存活率和D形幼虫发生率总体呈下降趋势,遗传失活的精子与正常卵子受精后其胚胎发育至D形幼虫前期停止。经紫外线照射...