基于形态学和DNA条形码探讨“南鲳”物种有效性

众所周知鲳科鱼类均为重要的经济种类,在中国沿海仅见鲳属(Pampus)鱼类分布,而低鳍鲳属(Peprilus)和真鲳属(Stromateus)则分布于大西洋和东太平洋。本研究采用形态学和DNA条形码技术对海南、广西和广东水产品市场上出现的“南鲳”进行物种鉴定,同时下载该物种的COⅠ基因同源序列进行比较分析,并纠正GenBank中的错误序列。研究结果显示“南鲳”学名实为中间低鳍鲳(Peprilus medius),为冰冻进口货品,在我国沿海无该物种的分布报道,其与国内已知的6种鲳属鱼类在鳃耙、背鳍、臀鳍等外部形态上存在明显的差异,在COⅠ基因水平上与鲳属鱼类遗传距离为0.138,分化于中新世晚期。本研究可为水产品市场种类鉴定提供借鉴,也为鲳科鱼类的研究和海洋鱼类分类学提供基础数据和参考。     

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...     

海洋生物DNA条形码研究现状与展望

海洋生物种类多样,分布广泛,具有复杂性、多样性和趋同性等特点,为了对物种进行更快速、准确地鉴定,急需在传统形态分类学基础上,建立并结合便捷准确的分子鉴定手段。DNA条形码提供了可信息化的分类标准和有效的分类学手段,已成为近年来分类学与生物多样性研究中重要的技术依托。本文概述了DNA条形码当前的发展现状与趋势,并介绍了DNA条形码技术在主要海洋浮游植物(红藻、褐藻、绿藻、硅藻、甲藻)、无脊椎动物(海绵动物、刺胞动物、甲壳动物和软体动物等)和鱼类中的研究进展,以及不同条形码基因针对于不同生物类群的有效性和适用性,指出了目前条形码技术在各海洋类群中存在的主要问题,并对未来的相关工作做了展望,希望为今后我国的海洋生物DNA条形码研究提供理论基础。     

基于DNA条形码的长江口鱼类浮游生物形态分类研究

DNA条形码技术是利用一段较短的DNA序列实现快速准确物种鉴定的工具。本研究采用传统形态学和DNA条形码技术相结合,对长江口及其邻近水域的鱼类浮游生物种类进行准确鉴定,并对部分种类仔稚鱼进行了形态学描述。结果表明：2016年春、夏季和2017年夏季共获得鱼类浮游生物55种,隶属9目19科。其中鲈形目种类数最多,为35种。鱼类浮游生物在类群上季节变化不明显,但在种类上有明显的季节变化。仅有凤鲚(Coilia mystus)、日本鳀(Engraulis japonius)和小黄鱼(Larimichthys polyactis)在春、夏季同时出现。首次描述了龙头鱼(Harpadon nehereus)仔、稚鱼阶段的形态特征;对龙头鱼、前鳞龟鰉(Chelon affinis)、四指马鲅(Eleutheronema tetradactylum)、少鳞鱚(Sillago japonica)和日本须鳎(Paraplagusia japonica)这5种仔、稚鱼的可量性状、鳍的发育、黑色素的分布等情况进行了描述,绘制了仔、稚鱼形态图。以上研究可为河口鱼类育幼场的研究提供科学依据,也为鱼类早期发育阶段分类资料的积累探索新途径。     

浙江海域天竺鲷科鱼类新纪录种—黑边银口天竺鲷(Jaydia truncata)形态特征与DNA条形码研究

2015年10月中旬,在浙江省舟山市近海的渔业资源调查中采集到天竺鲷标本,经鉴定发现其中黑边银口天竺鲷为浙江新纪录种。本文对采集的6尾黑边银口天竺鲷标本拍摄照片并开展形态特征和DNA条形码研究,对该种的拉丁学名进行校正。黑边银口天竺鲷的有效学名为Jaydia truncata(Bleeker,1854)。具有以下主要形态特征:背鳍鳍条数VII+I,9,胸鳍鳍条数16—17;背前鳞数4;第一鳃弓总鳃耙数3—4+11—13,发达鳃耙数1+9—10,角鳃弓上的鳃耙数9;前鳃盖骨边缘锯齿状;第一背鳍上端黑色,第二背鳍及尾鳍边缘黑色,第二背鳍与臀鳍中部各具一黑色纵纹。本研究分析了浙江海域5种天竺鲷科鱼类的形态差异,并编制了检索表。测定了6尾标本的DNA条形码,结合Gen Bank中所有学名为Jaydia truncata及其同种异名的同源序列分析,发现所有个体明显分为两个分支,且两者遗传距离达到0.149,表明两分支可能为不同的有效种,Gen Bank中只有AB890050和JQ681488与本研究序列结果相近。     

中国沿海常见鲳属鱼类耳石形态研究

本研究采集中国沿海常见五种鲳属鱼类并对其耳石进行系统的多元形态学描述与比较。耳石形态在整体轮廓、边缘以及前部表现出明显的种间差异,展现了耳石形态的种内特异性。随着基因隔离与种间分化,鲳属鱼类耳石也逐渐变薄且基叶、翼叶更为发达。上述观察到的种间耳石形态差异与鱼体本身生活的水层以及温度联系紧密。96.6%的判别成功率也证明鲳属鱼类耳石显著的种间差异,表明形态学方法可以作为鲳属鉴别的有效手段之一。     

DNA条形码技术在鱼类浮游生物鉴定中的应用及研究展望

鱼类浮游生物的准确鉴定是鱼类浮游生物研究的基础。传统的基于形态特征的鉴定不仅费时费力,而且由于缺乏足够信息,鉴定存在困难,导致物种多样性被低估。为了对物种进行准确、快速地鉴定,急需在传统形态分类学基础上,建立并结合便捷准确的分子鉴定手段。DNA条形码技术是利用一段较短的DNA序列实现快速准确物种鉴定的工具,就像在商店里扫描仪读取条形码那样,对每一种生物也能通过快速分析其DNA中的一小段(线粒体细胞色素c氧化酶I亚基,mt COI)加以识别。DNA条形码提供了可信息化的分类标准和有效的分类学手段,已成为近年来生物类群的研究热点。文章介绍了DNA条形码的概念、原理、工作流程及其优点,分析了其在鱼类浮游生物鉴定中的应用可行性,并展望未来鱼类浮游生物学发展的前景:分子技术鉴定鱼类浮游生物相关规范标准的建立,传统形态鉴定与分子方法相结合的分类学研究,以及鱼类浮游生物的生态学研究。     

东海几种鳗鲡目鱼类DNA条形码研究

本研究采集了分布于中国东海的前肛鳗(Dysomma anguillaris)、短尾蛇鳗(Ophichthus brevicaudatus)、艾氏蛇鳗(Ophichthus evermanni)、海鳗(Muraenesox cinereus)、黑尾吻鳗(Rhynchoconger ectenurus)、微鳍新鳗(Neenchelys parvipectoralis)、大头蚓鳗(Moringua macrocephalus)、梅氏美体鳗(Ariosoma meeki)和星康吉鳗(Conger myriaster)9种鳗鲡目(Anguilliformes)鱼类,采用PCR技术扩增了线粒体COI基因片段序列, 结合GenBank数据库中下载的5种鳗鲡目鱼类同源序列, 分析比较了序列组成和差异, 并以光海鳝(Muraena argus)和细点海鳝(Muraena augusti)为外群, 基于最大似然法构建了鳗鲡目中6科11属14种鱼类的系统发育树, 探讨了该类群鱼类的系统进化关系。结果显示:72条序列共检测到43种单倍型, 4种碱基含量分别为27.4%(T)、28.2%(C)、25.8%(A)、18.6%(G), 平均A+T含量(53.2%)高于G+C含量(46.8%), 表现出明显的碱基组成偏好性。基于K2P(Kimura 2-parameter)模型计算得出不同种间的平均遗传距离为0.218 8, 不同属间的平均遗传距离为0.225 0, 不同科间的平均遗传距离为0.232 7, 分类阶元越高, 遗传距离越大。系统进化树显示蛇鳗科物种都能够形成独立的分支, 并得到有效的区分, 而其他类群存在混杂现象。以上结果表明, 由于鳗鲡目鱼类种类多且分布广,线粒体COI基因只适用于较低分类阶元(如科内属间、属内种间)间的物种鉴定, 该类群鱼类系统发育关系还有待于结合多种DNA条形码进行深入探讨。     

我国近海裸胸鳝属鱼类新记录种——黏裸胸鳝（Gymnothorax mucifer）形态与分子系统学研究

本研究于2020-2021年在福建省厦门市近海水产市场采集到4尾黏裸胸鳝样品,为中国大陆近海新记录种。此前,该物种仅在澳大利亚、夏威夷等地有分布记录,并被认为是蠕纹裸胸鳝同种异名。本研究对采集的黏裸胸鳝进行详细形态特征分析,结合DNA条形码COI基因进行分子鉴定及系统进化关系分析。黏裸胸鳝的主要鉴别特征：体为黄棕色,头部前端淡紫色,身体布满细长、稀疏、弥散的树枝状不规则棕色斑纹,斑纹颜色靠近尾部加深加粗,形成网格状;臀鳍边缘白色,近尾部变为一列连续白色斑点;全长为体长的1.01倍,为头长的8.00～8.39倍;上颌齿每侧8～10个,下颌齿每侧14～20个,中央齿细长,均为单行;总椎骨数为117～139,平均椎骨式为6-47-130。基于COI基因分析,黏裸胸鳝与蠕纹裸胸鳝的遗传距离为0.074,大于Herbert设定的2%(0.020)作为区分不同物种最小遗传距离,表明两者应为不同的物种。形态上,两种裸胸鳝也存在差异特征,如黏裸胸鳝的斑纹较细长、稀疏、不明显,头部前端偏淡紫色,臀鳍白色边缘近尾部断裂成系列连续白色斑点;蠕纹裸胸鳝的斑纹粗大明显、颜色较深,头部前端黄白色,臀鳍白色边缘连续...     

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。     

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.     

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.     

Levels and patterns of genetic variation in Pampus minor:Assessment of a mitochondrial DNA control region sequence

Pampus minor is an important commercial fish. Due to the similarity of external morphological characteristics among the genus Pampus species, P. minor has often been identified as the juvenile group of both P. cinereus and P. argenteus. While little genetic background on this species is known, this study was based on control region sequences and provided the first evaluation of the genetic signature of 264 individuals of P. minor from 11 populations along the coasts of China and Malaysia. The results indicate high genetic haplotype diversity and low nucleotide diversity in this species. Additionally, two differentiated haplotype lineages were identified in the P.minor populations. However, phylogenetic structures corresponding to the geographical locations were unable to be established. Analysis of molecular variance identified a vast majority of the genetic variation occurring within populations. F-statistic test value(FST) of pairwise indicated that great differences existed between the Chinese and Malaysian P. minor populations. For the Chinese populations, the genetic differences were insignificant with the exception of the Xiamen population, which is a marginal population. During the late Pleistocene, a population expansion of P. minor occurred. These expanded populations originated from the glacial refugium in the South China Sea and then rapidly occupied and adapted to their new habitat. The results of this study provide genetic information for ensuring the protection and management of P. minor resources.     

短小拟五角蟹的分布新记录

2022年9月作者在浙江宁波象山港海域采集到短小拟五角蟹(Paranursia abbreviata),由此短小拟五角蟹的地理分布记录从中国的海南岛、广东、香港和福建扩展到东海北部,即浙江北部的象山港。本文对其形态特征进行了详细的描述,并讨论了其与历史记载的外形差异。由于象山港冬季水温较低,推测短小拟五角蟹在象山港的出现可能是台湾暖流携带幼体定居导致的偶然现象。     

中国海盘海牛科(腹足纲, 裸鳃目)一新记录属和一新记录种

2021年3月在大连进行潮间带生物调查时采集到2个裸鳃类标本,经鉴定发现为欧氏针盘海牛Diaulula odonoghuei (Steinberg, 1963),为中国新记录属和新记录种。在研究中对其进行了活体拍照,利用光学显微镜和扫描电子显微镜对其外部形态和内部解剖特征进行了观察。此外,测定了这两个标本的COI、16S rRNA和H3基因序列,将测得的基因片段与GenBank数据库中的针盘海牛属Diaulula的同源序列进行对比分析,构建了针盘海牛属的系统发育树。欧氏针盘海牛在我国沿海的发现,进一步丰富了中国海软体动物的物种多样性研究。     

Genetic and morphological divergence in the purple sea urchin Paracentrotus lividus (Echinodermata, Echinoidea) across the African Mediterranean coast

The present investigation focuses on population structure analysis of the purple sea urchin Paracentrotus lividus across the African Mediterranean coast, with the main aim of assessing the influence of the Siculo-Tunisian Strait on gene flow disruption in this highly dispersive echinoid species. For this purpose, patterns of morphological and genetic variation were assessed among its populations from the western and eastern Mediterranean coasts. A total of 302 specimens from seven Tunisian sites were collected and examined for morphometric variability at twelve morphometric traits. Concordant results, inferred from CDA (canonical discriminant analyses), pairwise NPMANOVA (non parametric multivariate analysis of variance) comparisons and MDS (multidimensional scaling) plot, unveiled significant inter-population differences in the measured traits among the studied populations. Furthermore, the combined use of the one way ANOSIM (analysis of similarities) and the Discriminant/Hotelling analysis allowed unravelling two morphologically differentiated groups assigned to both western and eastern Mediterranean basins. The SIMPER (similarity percentages) routine analysis showed that total dry weight, test diameter and spine length were major contributors to the morphometric separation between locations and between groups. Pattern of phenotypic divergence discerned in P. lividus across the Siculo-Tunisian Strait is interestingly in congruence with that inferred from the genetic investigation of the purple sea urchin populations from the same region based on the analysis of the mtDNA COI (cytochrome oxidase I) gene in 314 specimens from nineteen locations covering a wider geographic transect, streching westward to the Algerian coast and eastward to the Libyan littoral. The specific haplotypic composition characterizing each Mediterranean basin, as inferred from the minimum spanning network, confirmed the geographic partioning of genetic variation, as revealed by F-statistics and AMOVA (analysis of molecular variance) analyses, yielding significant genetic differentiation between eastern and western Mediterranean populations. The newly detected phylogeographic patterns, observed for the first time in P. lividus throughout the explored distribution range, suggest the involvement of different biotic and abiotic processes in shaping such variation, and provide evidence that a large and geographically exhaustive dataset is necessary to unveil phylogeographic structure within widespread marine species, previously cathegorized as panmictic in part of their distribution range.