浮游软体动物长角螺属(Clio)种类的分类学和谱系地理学

许多终生浮游软体动物接近全球分布或呈环球分布,是海洋酸化和谱系地理学研究的良好材料。本文以西北太平洋和北印度洋的长角螺属(Clio)种类为材料,通过测定其线粒体COI基因(mtCOI,55条)和核18S rRNA基因(9条)序列,结合数据库中已有的序列,对该属进行了分类学和谱系地理学研究。结果表明,矛头长角螺(C. pyramidata)和尖棘长角螺(C. cuspidata)在mtCOI基因系统树中均形成4个明显分化的谱系分支,分别为支系A–D和支系E–H。矛头长角螺的支系A为全球分布,中国海及邻近海域可能仅有支系A的存在,支系B、C和D分布于特定海域。尖棘长角螺也存在明显的谱系地理结构,西北太平洋北赤道流南北两侧存在2个不同的支系,分布于吕宋海峡的支系E为一新的谱系分支。各支系内mtCOI基因的K2P遗传距离在0～0.026之间,支系间的遗传距离在0.031～0.089之间。膨凸长角螺(C. convexa)和曲形长角螺(C. recurva)没有明显的地理遗传分化。18S rRNA基因支持支系D为独立种,但不支持其他支系的划分。矛头长角螺和尖棘长角螺内部可能存在隐存多样性。洋流可...     

中国沿海常见蜑螺科贝类的DNA条形码

DNA条形码不仅为物种鉴定提供了有效方法,而且也有助于分类学和生物多样性研究。本研究旨在探讨将COI和16S rRNA基因序列应用于中国沿海蜑螺科贝类物种鉴定的可行性,获得了该科3属7种贝类61个个体的COI和16S rRNA基因序列。基于COI基因序列的种内遗传距离为0.00—1.29%,平均为0.67%;属内种间遗传距离为4.62%—19.25%,平均为13.02%;基于16S rRNA基因序列的种内遗传距离为0.00%—0.48%,平均为0.23%;属内种间遗传距离为2.47%—8.48%,平均为6.37%。两种基因序列在所研究的蜑螺中,种内遗传差异均小于种间遗传差异,存在明显的条形码间隙,所有物种在系统发生树上都表现为独立的单系群。结果表明,线粒体COI和16S rRNA基因序列可以作为DNA条形码标准基因对蜑螺科贝类进行有效地物种鉴定。     

红藻DNA条形码分析技术研究进展

DNA条形码是指利用一段相对较短的标准DNA片段,对物种进行识别和鉴定。目前该技术在动物、植物物种鉴定领域已经得到了广泛的研究和应用。在藻类的研究中,尚未确定一条统一的标准条形码基因,现阶段都是使用2条或2条以上基因序列来完成物种鉴定。对于形态多样、种类繁多的海洋红藻,常用的DNA条形码基因有COI基因(Partial cytochrome c oxidase I gene)、UPA基因(Partial 23SrRNA gene,universal plastid amplicon)、LSU基因(Partial 28SrRNA gene)和rbcL基因(The large subunit of ribulose-1,5-bisphosphate carboxylase)等,这些基因中2个或3个基因的互补运用准确有效地提高了红藻的鉴定准确率,尤其是COI基因的种间差异大足够区分相近物种。本文在概述条形码的原理及其标准的基础上,阐述了红藻DNA条形码鉴定研究的新进展以及常用几种基因片段的优缺点,并对条形码在红藻鉴定中存在的问题进行了分析,对其应用前景进行了展望。     

中华钩线虫的DNA条形码研究及分类意义探讨

海洋线虫是海洋底栖生物中数量上最丰富的类群,在海洋生态系统中起着重要的作用。对海洋线虫进行种类鉴定是线虫研究中最重要的工作之一。目前,海洋线虫的鉴定主要采用形态学的分析方法,但是这种方法往往费时费力,对于如此丰富的物种,急需新的鉴定方法。作者以黄海潮间带自由生活线虫优势种——中华钩线虫(Oncholaimus sinensis)为例,在形态学分类的基础上,将DNA条形码技术引入线虫的鉴定中,探讨了线粒体细胞色素C氧化酶第一亚基(mt COI)基因序列、28S r DNA序列的D2D3区以及18S r DNA序列的部分序列作为DNA条形码在中华钩线虫中的适用性。结果表明,18S r DNA序列可作为该种线虫的DNA条形码,为海洋线虫的DNA条形码研究提供了很好的借鉴。目前,利用DNA条形码技术对海洋线虫进行鉴定的报道在国内还属空白,本研究将是海洋线虫分类学研究的很好补充。同时,对于了解该海域海洋线虫多样性及群落分布格局,开展海洋环境监测,进而对海洋的底质环境状况进行健康评价具有十分重要的科学意义。     

DNA条形码技术在鲻科鱼类鉴定中的应用

本研究选用DNA条形码的通用基因片段,采用特异性扩增测序及与GenBank已有序列结合分析的方法,进行了鲻科(Mugilidae)6属17种鱼类的COI基因片段的序列比较、分子树构建和系统进化研究。研究表明,在所得的17种鲻科鱼类共有的555bp COI基因片段中平均GC含量为46.9%。其中,第二密码子位点含量最高(49.8%~56.2%),平均54.9%;第一密码子变化范围最大(31.9%~48.6%),平均42.9%;第三密码子差别不显著(42.5%~43.4%),平均42.7%。依据Kimura-2-parameter模型(K2P),17种鲻科鱼类种内遗传距离的平均值为0.004 5,种间遗传距离为0.191,是种内遗传距离的42倍。在分子系统树上,所有物种均呈单系,5属为独立分支,只有隶属于梭属(Liza)的尖头梭(Liza tade)与莫鲻属(Moolgarda)聚类,表现出与传统形态学分类不一致的结果。本研究结果验证了线粒体COI基因作为DNA条形码对鲻科鱼类进行物种鉴定的有效性,可用于辅助探讨鲻科科下属、种分类阶元系统发育问题。     

小球藻属DNA条形码的鉴定研究

为探讨小球藻鉴定的新方法以及评估小球藻属 DNA 条形码候选序列的鉴定作用,文章对20株小球藻的线粒体基因细胞色素C氧化酶亚基Ⅰ(COI)、核基因组rDNA的内转录间隔区(ITS)以及核酮糖1,5-二磷酸羧化酶(rbcL)3个片段进行PCR扩增测序,并对各序列进行生物信息学分析。研究结果表明：COI、ITS和rbcL序列在20株小球藻中的扩增和测序效率均呈阳性,扩增成功率分别为76%、83%和70%;经评估,COI、ITS和rbcL序列作为DNA条形码在小球藻分类鉴定中均不适合较低的分类单位,但可参考动植物的分类方法,将多个片段组合应用,从而筛选不同进化级别的DNA条形码。     

中国梭螺科一新记录属

本研究记述了采集自海南陵水海域的梭螺科(Ovulidae)一新记录属：舟梭螺属(Naviculavolva)及一新记录种：折唇舟梭螺(N.deflexa),对标本进行了形态描述并附标本照,且通过比对样品的COⅠ基因序列证实了形态鉴定的准确性。本研究为梭螺科分类与研究提供参考资料,丰富了我国海域梭螺科物种的多样性,并使得折唇舟梭螺的分布范围延伸至海南省海域。     

DNA条形码技术在仔稚鱼鉴定中的实践

对物种准确鉴定是进行生物多样性研究的前提。为提高物种鉴定的准确性,文章采用DNA条形码技术结合形态学对长江口仔稚鱼样品进行种类鉴定。结果表明,44个样品中,通过线粒体COI基因的分析鉴定有17个种、1个属;部分形态上难以鉴定的种类,如虾虎鱼科的矛尾虾虎鱼、斑尾刺虾虎鱼、睛尾蝌蚪虾虎鱼、褐吻虾虎鱼和斑点竿虾虎鱼,可通过DNA条形码实现有效鉴定;形态较为相似的种类,通过DNA条形码可鉴定到种,淡水鲤科的有青鱼、鲢、鳊,鲻科的有龟鮻和前鳞龟鮻。利用DNA条形码技术结合形态进行仔稚鱼种类的鉴定,能提高仔稚鱼鉴定的精确度。     

中国大陆近海菖鲉属鱼类新记录种——三色菖鲉(Sebastiscus tertius)的形态特征与DNA条形码研究

2018年4月在浙江省舟山近海海域的渔业资源调查中采集到19尾菖鲉属鱼类标本,经鉴定发现为三色菖鲉(Sebastiscus tertius),为中国大陆近海海域的新记录种。本研究对采集到的19尾三色菖鲉标本拍摄照片并对其进行形态特征以及DNA条形码研究。三色菖鲉具有以下主要形态特征:背鳍数ⅩⅡ-12,胸鳍数18—19,腹鳍数Ⅰ-5,臀鳍数Ⅲ-5;第一鳃弓总鳃耙数6—7+13—17;前鳃盖骨有5枚硬棘,鳃盖骨后沿有2枚硬棘;眶前骨下后角有1枚硬棘;胸鳍处有一明显黑斑。测定了19尾标本的12S rRNA和COI序列,结合GenBank中所有学名为Sebastiscus tertius的同源序列进行分析,发现12S rRNA序列完全一致,可作为该物种的微型DNA条形码,而COI序列中所有个体明显分为两个分支,两者遗传距离达到0.044,表明两分支可能为不同的有效种。COI同源序列分析结果显示, GenBank中学名为Sebastiscus tertius的鱼种与褐菖鲉(Sebastiscus marmoratus)聚为一支,表明GenBank中的Sebastiscus tertius存在错误...     

中国木叶鲽属鱼类一新纪录种

2008年8月,在浙江舟山附近海域(东海)采集到4尾木叶鲽属鱼类,经鉴定为中国鱼类新纪录种--长木叶鲽,其主要特征为:体棕褐色,有眼侧和无眼侧颢上枝前端有小分枝,鳞片小但不狭长,背鳍鳍条数72～74(73.3);臀鳍鳍条数56～59(57.5);有眼侧胸鳍鳍条数8～10(8.8),无眼侧胸鳍鳍条数为8;有眼侧与无眼侧腹鳍鳍条数均为6;尾鳍鳍条数19～20(19.5),脊椎骨数为36;鳃耙数为3+7.基于线粒体COI基因片段得到的长木叶鲽与角木叶鲽的K2P遗传距离为9.4%,2种间遗传分化达到了种间水平.     

褐藻纲海带目Laminaria属和Saccharina属基于三个分子标记的系统发育分析

In the past two decades, many studies have focused on the classification within genus Laminaria, ultimately trying to divide it into two subgroups or genera: Laminaria and Saccharina. A significant debate still surrounds the question of its division, as the conflicting phylogenetic hypotheses that have resulted from the classification studies are based on different taxon sampling, molecular markers, or analysis methods. It is aimed at elucidate the molecular phylogeny within Laminaria and Saccharina. The nine species of Laminariales are sampled from northern Asia and Europe, and 23 new sequences in the nuclear, plastid, and mitochondrial genomes are determined to identify their taxonomic status. The phylogenetic analyses of 71 species are performed, including representatives from six of the seven families of the order Laminariales, based on three separate data sets. An evidence is provided to strongly support a clear split that maintains the two recognized genera, Laminaria and Saccharina, with Laminaria appearing to be the ancestor group. Further, analyses indicate that all taxa in Saccharina and Laminaria did not form a monophyletic lineage, instead Laminariaceae and Lessoniaceae grouped together interlacedly, and Costariaceae appeared as the sister taxon of the Lessoniaceae–Laminariaceae clade. In the phylogenetic analysis, mitochondrial c oxidase I(COI) sequences appeared to be the most credible molecular marker which was more befitting than nuclear encoded internal transcribed spacer(ITS) and plastid encoded rbcL for establishment of Laminariales systematics. It is the most comprehensive phylogeny of the order Laminariales, and contributes to an enhanced understanding and estimation of the phylogenetic relationships for the economically important seaweeds, Laminaria and Saccharina.     

基于线粒体COI 基因的毛蚶群体遗传多样性

采用PCR技术,扩增了大连、乳山、烟台、舟山4个毛蚶(Scapharca subcrenata)地理群体共38个个体的线粒体COI基因部分序列,并分析了4个毛蚶群体的遗传多样性和系统发育关系。研究结果显示:38个毛蚶COI部分序列经处理得到长度均为625bp的基因片段,共分为30种单倍型;基于COI部分序列的分析结果,毛蚶4个地理群体总的变异位点为301个,多样性指数Pi为0.15048,平均核苷酸差异数为92.242,单倍型多样性指数S为241。聚类分析显示毛蚶大连群体、乳山群体和烟台群体具有高度的遗传多样性,3个群体交叉聚在一起,没有明显的群体分化;舟山群体单独聚为一支,与其他3个群体分化明显。研究表明,线粒体COI基因不能单独做为毛蚶大连、乳山和烟台群体的遗传标记,但可以作为毛蚶舟山群体的有效群体遗传标记,为线粒体COI基因在群体遗传学的应用提供了基础资料。     

Species delimitation in the green algal genus Codium (Bryopsidales) from Korea using DNA barcoding

Codium, one of the largest marine green algal genera, is difficult to delimit species boundary accurately based on morphological identification only. DNA barcoding is a powerful tool for discriminating species of seaweeds. The plastid elongation factor TU(tuf A) is considered as maker to perform DNA barcoding of green algal species than rbc L gene due to universality and rapid evolution rate. We conducted DNA barcoding application to Codium specimens from the Jeju Island, Korea to overcome the limit of morphological identification and to confirm the species diversity. As a result of applying tuf A marker, we newly generated fifty-five tuf A barcodes to resolve eight species. Tuf A marker exhibited 6.1%–21.8% interspecific divergences, wider than the gap of rbc L exon 1,3.5%–11.5%. Molecular analysis of rbc L exon 1 sequences of Codium revealed eight distinct species like tuf A analysis separated in five phylogenetic groups. DNA barcoding of the genus Codium using tuf A marker is more helpful to overcome the limit of morphological identification, and this is more potential to reveal cryptic species and to resolve the relationships among subspecies than rbc L analysis alone. The complement of tuf A barcoding and rbc L analyses including morphology for the genus Codium in the northwestern Pacific will give much more reliable achievement for discovering species diversity and resolving the phylogenetic relationships.     

西北太平洋深海海山的一种蛇尾新记录

Ophioleila elegans是一种非常少见的蛇尾。2014年大洋35航次中利用“蛟龙”号载人潜器在西北太平洋麦哲伦海山链的采薇平定海山水深1660m处采集到2株该种标本。该种首次发现于夏威夷海山相似的生境当中,本次采集到的样品是继首次发现之后的再次公开报道。本文中对该种的形态学特征,如腕部骨骼等,进行了更加详细的研究,同时基于COI序列进行系统发育分析。形态和系统发育分析的结果都表明,Ophioleila属与辐蛇尾科更加接近。     

Morphological and molecular evidence supports the first occurrence of two fishes,Siganus sutor(Valenciennes, 1835)and Seriolina nigrofasciata(Rüppell, 1829)(Actinopterygii:Perciformes), from marine waters of Odisha coast,Bay of Bengal,India

Marine ecosystems provide a wide variety of diverse habitats that frequently promote migration and ecological adaptation. The extent to which the geographic distribution of marine organism has reshaped by human activities remains underappreciated. The limitations intrinsic to morphology-based identification systems have engendered an urgent need for reliable genetic methods that enable the unequivocal recognition of fish species,particularly those that are prone to overexploitation and/or market substitution. In the present study, however, an attempt has been taken to identify two locally adapted fish species, Siganus sutor(Valenciennes, 1835) and Seriolina nigrofasciata(Rüppell, 1829) of order Perciformes, which happens to be the first record in Odisha coast,Bay of Bengal. The diagnostic characteristics of Siganus sutor are: dorsal fin XIII-10, anal fin VII-9, pectoral fin 15,pelvic fin II-3, while that of Seriolina nigrofasciata dorsal fin VI-I-35, anal fin I-17, pectoral fin 16, pelvic fin 5. All COI barcodes generated in this study were matched with reference sequences of expected species, according to morphological identification. Bayesian and likelihood phylogenetic trees were drawn based on DNA barcodes and all the specimens clustered in agreement with their taxonomic classification at the species level. The phylogeographic studies based on haplotype network and migration rates suggest that both the species were not panmitic and the high-frequency population distribution indicates successful migration. The result of this study provides an important validation of the use of DNA barcode sequences for monitoring species diversity and changes within a complex marine ecosystem.     

2022年夏季胶州湾致灾物种多棘海盘车的分子解析

多棘海盘车(Asterias amurensis)是海星纲的一个致灾物种,广泛分布于包括我国黄海、渤海海域、韩国、日本和俄罗斯海域在内的北太平洋海域,并作为入侵种分布于澳大利亚的塔斯马尼亚海域。多棘海盘车在多个海域发生过暴发性增殖导致的高密度、大规模聚集,对海洋生态系统和水产养殖造成严重的负面影响。自2006年以来在我国山东青岛海域多次发生大规模多棘海盘车聚集,对贝类养殖造成重大经济损失。日本、俄罗斯、澳大利亚等海域的多棘海盘车遗传多样性研究取得了显著进展,然而,迄今为止鲜有针对我国海域致灾物种多棘海盘车的分子生物学分析,阻碍了我国海域种群与全球种群间遗传进化关系的比较分析。研究重点分析了于2022年7月在胶州湾发生多棘海盘车暴发时采集的12个样本,系统组装了其线粒体基因组、核糖体基因簇以及多种通用分子标记。基于线粒体基因组和核糖体基因簇的系统发育分析表明多棘海盘车具有较高的种内遗传多样性。基于线粒体基因组的蛋白编码基因进行分化时间估算,多棘海盘车的种内遗传分化可能发生在1.7～4.2 Ma,表明多棘海盘车的扩布可能较早发生。与线粒体基因组相比,多棘海盘车核糖体基因簇序列在种内极为保守...     

Genetic diversity and population structure of three dominant myctophid fishes (<Emphasis Type="Italic">Diaphus theta,Stenobrachius leucopsarus</Emphasis>, and <Emphasis Type="Italic">S. nannochir</Emphasis>) in the North Pacific Ocean

We compared the genetic diversity of three dominant myctophid fishes in the North Pacific Ocean that have different diel vertical migration patterns on the basis of the nucleotide sequences of the mitochondrial gene for cytochrome b. No genetic structure was detected for each of these three species. The genetic diversity progressively increased for Diaphus theta, a diel migrant species showing clear diel vertical migration; Stenobrachius leucopsarus, a semi-diel migrant, in which only part of the population migrates vertically; S. nannochir, a non-diel migrant. All three species were suggested to have experienced a recent, sudden population expansion. Interspecific differences in genetic diversity might be attributable to differences in the degree of population size reduction during the glacial periods; this degree in turn corresponds to the energy demand of the fishes.     

西太平洋巴特柔鱼放射性核素活度浓度测量与辐射剂量评估

为进一步认识日本福岛核事故对海洋生物体内放射性水平及公众辐射剂量的影响,本研究对日本福岛核事故后采集自西北太平洋的巴特柔鱼(Ommastrephes bartramii)器官组织中总α、总β放射性活度进行了测量分析,结果表明:巴特柔鱼各内脏器官中总α和总β的活度浓度明显高于其他软组织,其中以肝脏最为明显;巴特柔鱼各内脏器官的总β的活度浓度明显高于总α活度浓度。除此之外,利用国际上公开发布的海洋环境核素活度浓度水平,对日本近岸以及西太平洋公海的巴特柔鱼进行了3个时间段的辐射剂量评估。评估结果表明,根据公开数据,日本近岸受影响严重且时限较长,西太平洋公海区域受影响程度不大,时限亦不长。