厚蟹线粒体16S rRNA基因序列分析及系统发育研究

对我国沿海天津厚蟹6个群体、侧足厚蟹4个群体、伍氏仿厚蟹2个群体和日本仿厚蟹1个群体的线粒体16S rRNA基因片段进行了序列测定;结合从GenBank下载的其他厚蟹序列,分析了厚蟹分子系统发育关系.除天津厚蟹丹东群体的2个个体16S rRNA基因片段长度为526 bp外,其他天津厚蟹和侧足厚蟹均为525 bp;除日照...     

东海三叶原甲藻的形态特征及其ITS序列分析

报道了首次分离于东海海域的三叶原甲藻(Prorocentrum triestinum Schiller)藻株号(LAMB100721),通过利用光学显微镜、荧光显微镜、扫描电镜及分子生物学方法,对其形态特征、显微结构和分子系统进化进行了详尽描述和鉴定。细胞长卵形或披针形,后端细长且尖,前端圆,最宽部位于细胞中央。顶刺长而显,三角状。壳面光滑,无刺或突起物。刺丝胞孔稀疏而不规则地分布于壳面边缘。叶绿体无具体形状,分布于整个细胞中,细胞核球形,位于中下部。老化的细胞可见细胞边缘的间接带,且间接带表面光滑。细胞长为19~25 μm,平均值为(22.6±0.9)μm;宽为11~16 μm,平均值为(13.2±1.1) μm。所测目标藻株的rDNA ITS序列长度为569 bp,其中GC含量为47.6%。三叶原甲藻系赤潮种,为东海原甲藻春季大规模赤潮的伴随种。加强有害赤潮的预防和监测工作是减少危害的有效途径,而对赤潮原因种的准确识别和鉴定则是基础和关键。     

东海三叶原甲藻的形态特征及其ITS序列分析

报道了首次分离于东海海域的三叶原甲藻(Prorocentrum triestinum Schiller)藻株号(LAMB100721),通过利用光学显微镜、荧光显微镜、扫描电镜及分子生物学方法,对其形态特征、显微结构和分子系统进化进行了详尽描述和鉴定。细胞长卵形或披针形,后端细长且尖,前端圆,最宽部位于细胞中央。顶刺长而显,三角状。壳面光滑,无刺或突起物。刺丝胞孔稀疏而不规则地分布于壳面边缘。叶绿体无具体形状,分布于整个细胞中,细胞核球形,位于中下部。老化的细胞可见细胞边缘的间接带,且间接带表面光滑。细胞长为19~25 μm,平均值为(22.6±0.9)μm;宽为11~16 μm,平均值为(13.2±1.1) μm。所测目标藻株的rDNA ITS序列长度为569 bp,其中GC含量为47.6%。三叶原甲藻系赤潮种,为东海原甲藻春季大规模赤潮的伴随种。加强有害赤潮的预防和监测工作是减少危害的有效途径,而对赤潮原因种的准确识别和鉴定则是基础和关键。     

印度洋、西北太平洋龟螺属和小龟螺属种类的分类及DNA条形码鉴定

近来的研究表明,一些所谓的环球或环极地分布的广布种实际上包含着一些局限性分布的隐存种,物种多样性可能被低估。本文采用形态学和DNA条形码技术相结合的方式,对印度洋和西北太平洋海域的龟螺属(Cavolinia)和小龟螺属(Diacavolinia)的种类进行了分类学研究和物种鉴定。结果表明,线粒体16S rRNA基因数据不支持小龟螺属形态种的划分,分布于西北太平洋的D. grayi、D. vanutrechti、D. pacifica、D. elegans、D. angulosa等多个形态种可能属同一个种,即长吻小龟螺(D. longirostris)。COI基因数据也不支持钩龟螺(C. uncinata)亚种和变形的划分。许多形态特征不能作为种或种下分类单元的区分依据。钩龟螺、球龟螺(C. globulosa)和长吻小龟螺在COI系统树中均形成2个地理支系,其内部可能存在隐存种。西北太平洋海域长吻小龟螺的核基因组中存在线粒体假基因,对DNA条形码分析产生严重干扰。     

基于线粒体COI和16S rRNA基因的中国沿海相手蟹系统发育研究

相手蟹科的诸多种类因其形态极其相似成为方蟹总科分类中疑问较多的一个类群。通过对中国沿海相手蟹线粒体COI和16S rRNA基因序列进行分子系统发育分析,结果表明14种相手蟹COI和16S rRNA基因序列之间差异分别为5.7%～14.5%和1.5%～12.1%,均达到了种间差异水平。构建的系统发育树显示,14种相手蟹分别为独立有效物种,但分属于拟相手蟹属和近相手蟹属的4种拟相手蟹和3种近相手蟹,没有分别形成2个独立的支系,而是混合聚成一大支系。而属于螳臂相手蟹属的无齿螳臂相手蟹则首先与属于中相手蟹属的中华中相手蟹聚成一支,再与红螯螳臂相手蟹聚为一大支,表现出与形态分类的不一致。错综复杂的分子系统关系预示着相手蟹类为多系起源,也表明它们之间的种间关系乃至于属间关系尚有诸多问题有待进一步厘定。     

Patterns of genetic and morphometric diversity in the marbled crab (Pachygrapsus marmoratus, Brachyura, Grapsidae) populations across the Tunisian coast

The present study reports on population structure analysis of the marbled crabPachygrapsus marmoratus (Fabricius, 1787) from the Tunisian coast, an appropriate location to study biogeographical process...     

基于线粒体COI和16S rRNA基因研究3个地理群体黑龙江河蓝蛤的遗传多样性

采用通用引物对辽宁盘锦、辽宁大连、山东日照的3个地理群体黑龙江河蓝蛤(Potamocorbula amurensis)COI和16S r RNA序列进行扩增、测序分析,得到30条658bp的COI基因部分序列和27条450 bp的16S r RNA基因部分序列。其中COI和16S r RNA基因部分序列T、C、A、G和A+T的平均含量分别为45.4%和32.0%、13.5%和13.3%、20.7%和29.3%、20.4%和25.3%、66.1%和61.3%,AT含量高于GC含量,这与其他软体动物门动物的COI和16S r RNA的观测结果相近。COI和16S r RNA分别检测到了24个单倍型、43个核苷酸多态位点和9个单倍型、19个核苷酸多态性位点。AMOVA分析表明,3个群体间COI和16S r RNA部分基因总遗传分化系数分别为Fst=0.0090(P0.001)和Fst=0.0674(P0.001),群体内遗传分化远大于群体间、群体内存在较高的遗传分化。用NJ法构建分子进化树,3个地理群体的黑龙江河蓝蛤聚为一个族群,有少数个体和其他群体的个体聚在一起。     

基于S7核糖体蛋白基因部分序列的石鲈属鱼类分子系统进化关系

基于核DNA分子标记S7核糖体蛋白基因内含子1部分序列对分布于南海及美洲,非洲,大洋洲等周边海区的石鲈属鱼类11种,结合石鲈亚科其他属如仿石鲈属,异孔石鲈属,八带石鲈属及锯鳃石鲈属等21种鱼类的系统进化关系进行了研究分析。利用最大简约法以及贝叶斯法构建了系统进化树。进化树上,石鲈属的种类并没有形成一个单系,除了大斑石鲈,断斑石鲈等6种石鲈属鱼类聚成一石鲈属的分支外,其他石鲈属种类都位于进化树的不同位置。侧扁石鲈与八带石鲈聚在一起,大棘石鲈与异孔石鲈属聚在一起。红海石鲈与纵带石鲈与仿石鲈属的种类关系较近,佩氏石鲈最先分化出来,位于整个石鲈亚科分支基部。该分类关系与其各自的地理分布情况有着一定的联系。     

长心卡帕藻的形态结构观察与分子鉴定

通过形态构造观察和分子生物学相结合的方法,对栽培于我国海南的长心卡帕藻(Kap-paphycus alvarezii)株系XC-03(陵水)和麒麟菜族未知种株系LD-02(三亚)和LD-06(三亚)进行了鉴定,并与细齿麒麟菜(Eucheuma denticulatum)株系TSNY-05进行比较分析,结果表明:长心卡帕藻株系XC-03与麒麟菜族未知种株系LD-02和LD-06之间在分枝形状、长短、分布以及突起数量等藻体外部形态特征方面具有明显差异;它们的表皮、皮层和髓部细胞大小、皮层细胞层数和胶质层厚度等显微结构特征亦不相同,但分子鉴定表明该3个株系为同一个种,即长心卡帕藻。它们的ITS序列相似度为99.69%~99.85%,株系XC-03与LD-02之间仅有1个碱基转换和1个碱基颠换,株系XC-03与LD-06和株系LD-02与LD-06之间均分别仅有1个碱基转换和1个碱基颠换。长心卡帕藻与细齿麒麟菜的ITS序列相似度只有41.28%,两者差异较大。用RAPD,ISSR和ITS标记分析表明长心卡帕藻3个株系的两两间遗传距离分别介于0.122 9~0.153 5,0.068 8~0.112 4和0.002~0.005。用neighbor-joining和UPGMA聚类分析皆显示长心卡帕藻株系XC-03与LD-06的亲缘关系较近,与株系LD-02相对较远,而细齿麒麟菜单独列为一支。该研究为长心卡帕藻的种质鉴定和遗传育种研究提供了依据。     

三角褐指藻与抑食金球藻的竞争及化感作用研究

为探明三角褐指藻（Phaeodactylum tricornutum）对抑食金球藻（Aureococcus anophagefferens）是否具有化感作用,本研究以三角褐指藻和抑食金球藻为实验材料,研究了抑食金球藻在单培养和共培养条件下的生长情况及三角褐指藻培养滤液对其生长和叶绿素荧光参数的影响。结果表明:三角褐指藻对抑食金球藻有明显的化感抑制作用。在单培养体系中,抑食金球藻的生长曲线可用逻辑斯谛增长模型拟合,随着起始密度的增加,环境容量（K）逐渐减小,而抑食金球藻的种群瞬间增长率（r）、进入拐点时间及稳定期细胞密度均较为接近;当三角褐指藻与抑食金球藻以不同起始密度比共同培养时,抑食金球藻的生长均受到了显著地抑制（P<0.05）,但其抑制作用并未与三角褐指藻的密度呈明显的线性关系;滤液培养实验发现,10 mL和15 mL三角褐指藻培养滤液的加入可对抑食金球藻的生长产生显著影响（P<0.05）,对其叶绿素荧光参数没有影响（P>0.05）,25 mL和35 mL三角褐指藻培养滤液的加入可对抑食金球藻的生长和叶绿素荧光参数均产生显著的抑制作用（P<0.05）,叶绿素荧光参数F_v/F_m、ΦPSⅡ、alpha的值降低,Ik的值增加,PSⅡ受到损害。     

基于矢耳石形态的平鲉属鱼类鉴别及研究方法比较

分析了青岛胶州湾铠平鲉(Sebastes hubbsi)、花斑平鲉(Sebastes nigricans)和许氏平鲉(Sebastes schlegelii) 3种平鲉属鱼类的矢耳石形态特征,探究基于矢耳石形态鉴别3种鱼类的可行性.以8个耳石形状指标和37个椭圆傅里叶系数进行的单因素方差分析结果基本一致,显示铠平鲉与许氏平鲉的耳石形态差异最大,铠平鲉与花斑平鲉间差异最小.利用耳石形状指标、椭圆傅里叶系数、耳石形状指标与椭圆傅里叶系数相结合的方法,对3种鱼类的综合判别率均可达95%以上,判别分析散点图可明显将3种鱼类区分开.耳石长介于4~7 mm的3种鱼类小规格组与2龄及以上成鱼组耳石样本的判别结果一致,表明针对不同发育阶段平鲉属鱼类,基于耳石形态的种类鉴别均是可行的;仅使用椭圆傅里叶系数鉴别3种鱼类时综合判别率虽高达100%,但难以快速定位3种鱼类耳石形态间,在统计上差异显著而实际上微小的形状差异所在;而使用耳石形状指标和椭圆傅里叶系数联合判别,综合判别率亦高达99%,且能快速找出3种鱼类耳石在矩形趋近率、形态因子、面密度等的形态差异.研究表明,耳石形状指标和椭圆傅里叶分析联合判别是平鲉属鱼类种类鉴别及耳石形态差异分析的有效手段.     

两种体色三疣梭子蟹线粒体DNA 部分片段序列的比较分析

三疣梭子蟹(Portunus trituberculatus)作为重要的海洋经济动物,其常见体色(头胸甲)为茶绿色。近年来在我国沿海海域海捕三疣梭子蟹中开始出现体色为紫色的一类梭子蟹,除体色不同外,二者表型特征并无显著差异。对两种体色三疣梭子蟹不同个体的线粒体COI和16S rRNA基因进行了序列比对分析以判定二者之间的亲缘关系。结果发现COI和16S rRNA基因在两种体色三疣梭子蟹群体中的核苷酸序列一致性分别为99.87%和99.88%,表明紫色三疣梭子蟹并未发生亚种的分化,即紫色和茶绿色群体属于同一个种。并利用GenBank数据库检索了梭子蟹科其他15种海产蟹的16S rRNA序列,进行了序列同源性比对分析和分子系统学方面的探讨,为梭子蟹种属分类学研究提供分子依据。     

真鲷、黑鲷及其杂交子代的染色体构成与AFLP分析

采用核型分析和AFLP技术对真鲷、黑鲷及其杂交子代进行遗传差异分析。结果显示真鲷、黑鲷和杂交子代均含有48条染色体,核型分别为2n=2st+46t,NF=48、2n=4m+4sm+2st+38t,NF=56和2n=30m+8sm+2st+8t=48,NF=86,杂交子代核型与其父、母本种均不一致。两对AFLP选扩引物组合在真鲷、黑鲷和杂交子代中共扩增到278个条带,其中黑鲷特异性条带93条、真鲷特异性条带108条;杂交子代中分别出现了21条父本种(黑鲷)特异条带和67条母本种(真鲷)特异条带,另出现了15条非双亲条带。杂交子代与真鲷、黑鲷的遗传相似系数和遗传距离分别为0.113、0.350和2.180、1.050,表明杂交子代总体上更偏向于母本种。染色体核型和AFLP条带分析结果表明真鲷(♀)×黑鲷(♂)所获得的杂交子代为含有48条染色体的异源二倍体,且父、母本遗传物质在杂交中发生了部分重组,杂交子代表现出一定的偏母系遗传特性。     

Phylogeny of the cuttlefishes (Mollusca: Cephalopoda) based on mitochondrial COI and 16S rRNA gene sequence data

To clarify cuttlefish phylogeny, mitochondrial cytochrome c oxidase subunit I (COI) gene and partial 16S rRNA gene are sequenced for 13 cephalopod species. Phylogenetic trees are constructed, with the neighbor-joining method. Coleoids are divided into two main lineages, Decabrachia and Octobrachia. The monophyly of the order Sepioidea, which includes the families Sepiidae, Sepiolidae and Idiosepiidae, is not supported. From the two families of Sepioidea examined, the Sepiolidae are polyphyletic and are excluded from the order. On the basis of 16S rRNA and amino acid of COI gene sequences data, the two genera (Sepiella and Sepia) from the Sepiidae can be distinguished, but do not have a visible boundary using COI gene sequences. The reason is explained. This suggests that the 16S rDNA of cephalopods is a precious tool to analyze taxonomic relationships at the genus level, and COI gene is fitter at a higher taxonomic level (i.e., family).     

海月水母与海蜇不同幼体阶段的种间短期竞争研究

近年来海蜇池塘养殖业的兴起,补充了传统海蜇渔业捕捞生产的不足,推动了海蜇产业的迅猛发展。海月水母常常在海蜇养殖池塘中泛滥成灾,对海蜇养殖业常常造成很大损失,但关于海蜇与海月水母的种间竞争关系尚缺乏研究。本文对海月水母与海蜇的螅状体、碟状体及水母体幼体相互之间的短期生存竞争关系进行了初步观察研究。结果表明:除海月水母碟状体可以被海蜇螅状体所捕食外,各种类型的海蜇幼体基本不能对海月水母幼体造成危害。但海月水母螅状体及其水母体幼体对海蜇幼体均存在一定的危害性,尤其是对海蜇碟状体的危害性最为严重,在饵料缺乏的情况下海蜇碟状体可以100%地被捕食。随着海蜇幼体的生长,其对于海月水母危害的抵抗力逐渐增强。本研究显示海月水母相对于海蜇而言,在幼体种间竞争关系中处于绝对优势,这是海月水母时常暴发成灾的一个重要因素。在海蜇池塘养殖生产及苗种培育过程中,应当严禁混入海月水母幼体,以避免对海蜇养殖生产造成损害。     

旋链角毛藻对中肋骨条藻化感作用的影响因素

在已知旋链角毛藻对中肋骨条藻具有化感作用的前提下,进行了温度、生长时期、光照、细菌等因素对旋链角毛藻化感作用的影响、化感物质的萃取及粗提物对中肋骨条藻的化感效应检验等实验,研究了旋链角毛藻化感物质的性质、降解特点,并初步测定了化感物质的结构。结果表明:旋链角毛藻化感作用与生长时期有关,指数期滤液化感作用强于衰亡期;旋链角毛藻的化感物质在50℃以下保持稳定,光和细菌均引起旋链角毛藻化感物质的降解,且细菌降解强于光降解;旋链角毛藻的乙酸乙酯萃取物具有明显的化感活性,该萃取物在255～260 nm处有特征吸收峰。综合以上结果,推测旋链角毛藻化感物质可能为分解温度在50℃以上的易被光和细菌降解的苯衍生物,从极性上看较易溶于乙酸乙酯。     

缘管浒苔和羽藻氮、磷营养生理学研究

以无性繁殖系为材料,开展了缘管浒苔和羽藻对NO₃-,NH₄+和PO₄3-的吸收动力学、生长动力学研究。吸收动力学研究结果表明缘管浒苔和羽藻对NO₃- 和PO₄3-的吸收方式为主动运输,对NH₄+的吸收方式为被动扩散。缘管浒苔对NO₃-的最大吸收速率(V_max)、对NH₄+的吸收斜率都大于羽藻,说明缘管浒苔对高浓度的NO₃-和NH₄+具有更强的吸收能力。缘管浒苔吸收NO₃-和NH₄+的a值远大于羽藻,说明在低营养盐浓度时,缘管浒苔对NO₃-和NH₄+的亲和力更强。在PO₄3-的吸收中,羽藻的最大吸收速率(V_max)远大于缘管浒苔,说明羽藻对高浓度PO₄3-的吸收能力更强,但缘管浒苔的a值远大于羽藻,说明前者在低营养盐浓度时PO₄3-的亲和力更强。生长动力学研究结果表明,硝酸氮是促进两种海藻快速生长的最适宜氮源形式,氨氮更易促进藻体叶绿素的积累。在相同氮营养条件下,羽藻表现出比缘管浒苔更强的生长优势。     

Stage-V copepodites of Calanus sinicus and Calanus jashnovi (Copepoda: Calanoida) in mesopelagic zone of Sagami Bay as identified with genetic markers, with special reference to their vertical distribution

We applied genetic makers to identify Calanus species occurring in Sagami Bay, Japan, in order to investigate their vertical distribution in the upper 1000 m. First, interspecific genetic distances of three gene loci, mitochondrial small ribosomal RNA (srRNA), nuclear internal transcribed spacers 1 (ITS1) and 2 (ITS2), were estimated from morphologically distinguishable adult females of Calanus sinicus, Calanus jashnovi and Calanus pacificus that were collected from Sagami Bay, the Kuroshio Extension and the Oyashio region, respectively. The highest levels of interspecific genetic distance were observed in srRNA, followed by ITS1 and ITS2. The intraspecific genetic distances within C. sinicus were much lower than the interspecific genetic distances, indicating that DNA sequences in these loci are consistent with the morphological differences. This information was used as a criterion for species identification based on DNA sequence variation, and allowed us to identify the fifth copepodites (CVs) or younger stages of these species. Next, the vertical distribution of Calanus species was investigated in Sagami Bay in May 2006, on the basis of a stratified sampling in the upper 1000 m. By applying the genetic markers, 23 individuals comprising all copepodite stages were allocated into either C. sinicus or C. jashnovi, and the small- and large-sized CVs were identified as C. sinicus and C. jashnovi, respectively. The total abundance of C. sinicus was highest at 0-50 m and decreased with depth. On the contrary, CV individuals of C. sinicus were abundant not only in 0-50 m but also below 200 m with minimum occurrences in 150-200 m depth. C. jashnovi was much less abundant than C. sinicus and comprised of only CIV and CV which occurred in the upper 100 m and deeper than 50 m depths, respectively. The abundance of C. sinicus in the 1000-m water column of Sagami Bay was at a level comparable to that in shelf waters, suggesting the importance of off-shelf individuals in the biological production and organic transport in the respective areas.     

Distribution of amphipods Ischyrocerus on the red king crab, Paralithodes camtschaticus: Possible interactions with the host in the Barents Sea

The determination of costs and benefits experienced by crustaceans as a result of occupation by their symbionts has received increased attention from marine ecologists. However, the interactions between some important species and their associates remain unclear. We examined the distribution of amphipods in the genus Ischyrocerus on the red king crab Paralithodes camtschaticus, a commercially important species, in two areas of the Barents Sea. Ischyrocerus commensalis was found on 30.5% of crabs in Dalnezelenetskaya Bay (DZB) with the mean number per crab being 55.1, in Dolgaya Bay (DLB) these rates were 28.6% with 19.3 specimens per crab. Sympatric species Ischyrocerus anguipes was found on 13.5% of crabs in DZB with a mean of 7.3 individuals per host, in DLB it had much lower occurrence (1.3% and 1.5 specimens per host). There were no significant differences between proportions of male and female crabs infested by amphipods in both areas examined. Prevalence of amphipods was similar among years examined except for I. commensalis on small crabs (carapace length CL <90 mm) and I. anguipes on large crabs (CL >90 mm) in DZB. We found that I. commensalis and I. anguipes are not egg predators of P. camtschaticus in the Barents Sea, at least in summer. High numbers of I. commensalis occur in crab gills, and both mean intensity of the amphipods and their empty tubes increased with crab size. In the gills, I. commensalis predominated in the section nearest the mouth parts. Possible negative impacts for the hosts due to gills infestation are discussed. In contrast, I. anguipes were predominately found on the carapace and limbs of crabs and appears to be a less specific symbiont of P. camtschaticus. Both amphipod species seem to be commensals, however possible negative impacts for the host could not be excluded.     

Intraspecific diet shift in Talitrus saltator inhabiting exposed sandy beaches

Talitrid amphipods are the most abundant herbivores on exposed sandy beaches. Despite their important role as trophic intermediates between macrophytes and higher levels (i.e. insect and bird) of beach food webs, very little information is available on their feeding patterns. The main aim of this study was to investigate intraspecific differences in the feeding behaviour of Talitrus saltator. We tested the hypotheses that: (1) adult females and males showed different isotope signatures and therefore relied on different sources of food; and (2) patterns of variation of isotope signatures of juveniles differed from those of adult specimens, evidencing a diet shift during the development. We used stable isotope signatures and tested for differences upon the level on the shore, times of the year and beaches experiencing similar morpho-dynamic and environmental conditions. Finally, we investigated the trophic significance of macrophyte detritus in the diet of males, females and juveniles. Results showed that adult males had a more variable diet than females and juveniles (inferred from δ¹³C and δ¹⁵N values). Dual-isotope graphs suggested that Sargassum muticum and Cystoseira baccata wrack could be among the main food sources for both juvenile and adult stage.