线粒体COI基因条形码在鲿科鱼类物种鉴定中的应用

为了探讨线粒体COI基因作为DNA条形码在中国鲿科(Bagridae)鱼类物种鉴定中的有效性,以及系统发育中的适用性,本研究对4属11种鲿科鱼类进行PCR扩增,获得48条线粒体COI基因序列,同时从Gen Bank筛选获得8种鲿科鱼类的12条COI基因序列进行分析。19种鲿科鱼类的COI基因序列特征显示:长度为674 bp的COI序列片段平均碱基组成为24.82%A,30.44%T,27.10%C和17.64%G,碱基组成呈现明显的AT偏倚性(55.26%),具有硬骨鱼类的线粒体COI基因的碱基组成的典型特征。核苷酸位点中有变异位点226个,简约信息位点195个,单一信息位点31个,转换颠换比为3.35。19种鲿科鱼类的种内、种间和属间平均遗传距离分别为0.0041、0.1136和0.1268,种间遗传距离平均为种内遗传距离的27.7倍。在本研究中,鲿科鱼类所有的物种均形成单系,在物种鉴别上与形态学分类结果基本一致,然而鲿科的4个属中只有鱯属形成单系,黄颡鱼属、属和拟鲿属均未形成单系,其进化地位需要进一步研究。线粒体COI基因作为条形码可有效对鲿科鱼类进行物种鉴定,也为鲿科的系统发育提供了参考。     

黄渤海虾虎鱼DNA条形码分类体系构建

虾虎鱼(Gobiidae)是海洋底层食物链中的关键环节, 在生态系统的物质循环和能量流动中发挥重要作用。因虾虎鱼种类繁多、分布广泛、体型较小, 加之形态上存在不同程度的特化, 对形态学分类鉴定带来极大挑战。本研究对采自黄渤海的 73 尾虾虎鱼成鱼和幼鱼样本分别进行了形态学鉴定和 DNA 条形码分析, 依据形态学分类特征, 鉴定出 9 属 12 种; 而运用 DNA 条形码技术, 共鉴定出 13 种, 隶属于虾虎鱼科 10 属。可见, 在形态学鉴定经验不足或幼鱼形态学特征不明显的情况下, DNA 条形码可以有效地实现物种鉴定。同时, 系统整理了黄渤海虾虎鱼科记录种名录, 共计 29 属 47 种。依据物种名录, 从 BOLD 及 NCBI 数据库中筛选并下载了 18 属 29 种虾虎鱼科鱼类的 DNA 条形码, 与本研究鉴定出的 10 属 13 种虾虎鱼类合并分析, 构建了虾虎鱼科 26 属 42 种鱼类的系统关系树, 覆盖了黄渤海虾虎鱼科 89.36%的记录种, 通过对虾虎鱼科鱼类的分子系统发育关系与形态学分类地位进行确认和修订, 初步建立了黄渤海虾虎鱼科 DNA 条形码分类体系。本研究结果证明了 DNA 条形码技术对虾虎鱼物种具有较高识别效率, 有效弥补了传统形态学鉴定方法的缺点和局限, 丰富了虾虎鱼 DNA 条形码数据库, 完善了虾虎鱼 DNA 条形码分类体系, 为虾虎鱼的保护生物学、进化生物学及生物地理学等研究提供了科学依据。     

鲿科鱼类DNA条形码鉴定及系统进化研究

鲿科鱼类是一类高度多样化的类群,一些具有高度相似特征的物种难以通过形态进行识别,系统分类关系混乱,因此,选择一种更为有效、便捷的鱼类鉴定和分类方法十分必要。对5属36种共214条鲿科鱼类线粒体COⅠ基因序列进行分析,结果表明:36种鲿科鱼类的种内和种间平均遗传距离分别为0.016和0.158,种间平均遗传距离是种内平均遗传距离的9.875倍;大多数鲿科鱼类的种内遗传距离小于种间遗传距离,在种内、种间重叠较少,能形成的一定的DNA条形码间隙。ABGD结果将36个物种定义为25个运算分类单元,运算分类单元的划分与距离法结果大体一致,种间遗传距离较小的物种被划分为同一个运算分类单元,种内遗传距离较大的物种分化为多个运算分类单元。聚类树结果显示:黄颡鱼属、拟鲿属和■属的鱼类聚成一大支;鳠属和半鲿属中各有1个物种聚类到对方的分支中,这两属之间存在不完全的谱系分选。在本研究中,DNA条形码在大部分鲿科鱼类中能进行有效的鉴定,但对一些近缘种,条形码鉴定存在局限性,同时,COⅠ条形码也阐明了鲿科鱼类的系统进化关系,可为鲿科物种的系统分类提供科学依据。     

中国鲷科鱼类分类和系统发育研究进展

鲷科(Sparidae)隶属于硬骨鱼纲(Osteichthyes),鲈形目(Perciformes),鲈亚目(Percoidei).目前已报道世界有34属133种,广泛分布于大西洋、印度洋和太平洋的温带至热带水域,在世界海洋渔业中占有重要地位.虽然此类群系统学研究已有250多年历史,但是在亚科、属、种(亚种)等不同分类阶元上仍然存在颇多争议和疑问,有必要对其进行深人研究.本文总结了近百年来有关鲷科鱼类的分类和系统发育研究的文献资料,厘定了中国鲷科鱼类的有效物种和种名,修订了过去文献中出现的主要同物异名和错误的命名种,初步确认中国鲷科鱼类现有7属18种(或亚种);在探讨鲷科鱼类分类地位的基础上,指出了当前中国鲷科鱼类系统分类研究中存在的问题,展望了今后研究的方向,旨在为深入研究M科鱼类以合理保护和利用其资源提供参考依据.     

鲹科鱼类DNA条形码分类系统

为了验证 DNA 条形码在鲹科(Carangidae)鱼类物种鉴定和系统分类中的适用性, 本研究自测 6 属 7 种 17 条序列, 同时筛选了 BOLD 数据库中的有效序列, 共获得 25 属 95 种 273 条鲹科鱼类 DNA 条形码序列, 通过 BLAST 比对、遗传距离和系统关系树, 构建了鲹科鱼类的 DNA 条形码分类系统。结果表明: (1)鲹科鱼类属间、种间、属内种间和种内三级分类单元遗传距离的平均水平分别为 0.186、0.169、0.090 和 0.008, 种间平均遗传距离是种内平均遗传距离的 21 倍, 可见 DNA 条形码适用于鲹科鱼类分类鉴定; (2)运用 DNA 条形码技术可以识别出形态鉴定有误的物种, 表明 DNA 条形码可以弥补传统形态学鉴定的局限性, 可对鲹科鱼类形态学分类结果进行精准修正; (3)鲹科鱼类 DNA 条形码分析表明, BOLD 数据库中仍存在一定的“同种异名”和“异种同名”现象, 建议使用该数据库信息时应严格评估信息的准确性; (4)鲹科鱼类系统发生关系研究对物种的分类地位提出了新的见解, 即拟鲳鲹 (Parona signata)和镰鳍波线鲹(Lichia amia)亲缘关系较近, 支持将二者均归为鲳鲹亚科(Trachinotinae)。本研究旨在为丰富鲹科鱼类 DNA 条形码数据, 完善鲹科 DNA 条形码分类系统, 并为鲹科鱼类物种鉴定和系统分类提供分子证据。     

龙虾科物种线粒体基因组特征和系统发育分析

综合分析了龙虾科(Palinuridae) 13 个物种线粒体基因组的全序列, 发现其线粒体基因组的长度为 15470~ 16105 bp, A+T 含量为 62.63%~67.11%。Ka/Ks 分析表明, 龙虾科中的 10 个龙虾属(Panulirus)物种线粒体 13 个蛋白质编码基因(protein-coding genes, PCGs)的 Ka/Ks＜＜1, 显示出很强的纯化选择; 在差异位点的分析中, 发现 nd5、 nd4、rrnL 基因的差异位点比例较高, 是理想的分子标记, 可用于分析龙虾类不同群体之间的遗传多样性; 密码子偏好性分析表明, 被编码的氨基酸偏好性是相似的。同时, 本研究采用 ML (maximum likelihood)和 BI (Bayesian inference)方法构建系统发育树, 结果显示, 塔斯马尼亚龙虾属(Sagmariasus)物种最先开始分化, 而龙虾属单独聚为一支, 且与脊龙虾属(Linuparus)/游龙虾属(Puerulus)互为姊妹关系。贝叶斯分子钟估算结果显示, 龙虾科物种可能起源于三叠纪, 随后进一步分化为具有现代表征的龙虾种类。本研究旨在为快速鉴定龙虾科生物提供可靠的分子标记, 并为分析龙虾科物种遗传多样性提供理论依据。     

基于线粒体COI序列的DNA条形码在中国海口足类物种鉴定中的应用分析

中国海口足类动物区系具有丰富的物种多样性,是我国海洋底栖生物中的重要经济类群。口足类的属内种间鉴别特征有的极为相似,仅依靠传统的形态分类方法很难对近缘种和疑难种进行准确的鉴定。DNA条形码技术可以弥补传统形态学鉴定的某些局限,为物种鉴定提供了有效的工具。该研究探讨了利用线粒体COI序列对中国海口足类进行物种鉴定的可行性,共获得口足目4总科6科24属38个种的204条线粒体COI序列,与Gen Bank收录的14种42条口足类同源序列进行比对,结果显示口足类COI基因不存在碱基插入缺失现象,碱基组成偏倚明显,A+T含量(63.5%)显著高于G+C含量(36.5%)。基于Kimura双参数模型计算遗传距离,结果显示遗传距离随着分类阶元的增高而增大。同物种种内个体间的遗传距离变化范围在0%~3.91%,平均值为0.76%。同属内各物种间的遗传距离变化范围为6.55%~18.99%,平均值为12.91%。同科内不同属间的遗传距离变化范围为9.16%~23.32%,平均值为16.89%。不同科间的遗传距离变化范围为16.52%~26.6%,平均值为21.31%。由此可见,口足类COI基因的种间和种内遗传距离存在明显的间隙。基于COI序列构建的口足类邻接关系树显示所有包含大于1个个体的物种均可形成单系群,且节点支持率均为100%。本研究证明了COI序列作为DNA条形码标准基因在口足类物种鉴定中的有效性。此外,研究发现中国沿海分布的口虾蛄可能至少存在两个隐存种,实证了基于COI序列的DNA条形码技术能够用于口虾蛄隐存多样性的探究。     

南极鱼类DNA条形码及分子系统进化研究

本文采用COI基因兼并引物对15种36个南极鱼类DNA进行扩增测序,并结合Gen Bank已有序列进行联配分析,对南极鱼2科22属43种共97条COI基因片段(539 bp)进行序列比较和系统发生关系研究,探索了DNA条形码技术在辅助鱼类物种鉴定和分类中的适应性与可行性。结果分析表明,43种南极鱼科和鳄冰鱼科的鱼类COI基因的平均碱基组成为T:31.9%、G:18.3%、A:22.2%和C:27.6%,具有明显的碱基偏倚性。南极鱼类的种间平均距离为0.157,种内平均遗传距离为0.002,种间平均遗传距离是种内平均距离的79倍;系统分析结果显示,除南极小带腭鱼(Cryodraco antarcticus)和罗斯海小带腭鱼(Cryodraco atkinsoni)外,其余的鱼类皆能够形成独立的分支,且与形态学分支一致。由此可见,DNA条形码对南极鱼亚目鳄冰鱼科和南极鱼科鱼类能够进行有效的物种鉴定,基于COI基因所建的NJ(neighbor-joining)树对物种分类具有较为准确的辨识力。系统发生关系表明,DNA条形码可以对除南极小带腭鱼(Cryodraco antarcticus)和罗斯海小带腭鱼(Cryodraco atkinsoni)外的南极鱼物种进行鉴定,不仅可以作为形态学的辅助手段为南极鱼分类系统的必要补充和佐证,并且可以用于探讨南极鱼类近缘种的系统发育关系。     

基于CO I基因的福建近海部分仔稚鱼DNA条形码分析

为研究DNA条形码技术在遭遇瓶颈的仔稚鱼种类鉴定中的适用性,2015年7月17-20日采集福建近海仔稚鱼样品,并挑选80尾进行DNA条形码分析.获得仔稚鱼的CO I基因序列73条,鉴定仔稚鱼26种,隶属于7目22科25属,另有5个物种仅鉴定到属,2个物种仅鉴定到科.种内平均遗传距离为0.0023,属内种间遗传距离为0.1797,约为种内遗传距离的78倍,说明利用CO I基因可以进行有效的仔稚鱼物种鉴定.科内属间、目内科间的遗传距离分别为0.1937、0.2420,遗传距离随着分类阶元的提高而增大.在系统进化树的分析中,同一种类的不同个体都聚在一支,所有物种都分别聚为独立的一支,这些物种都能有效区分开来.以上结果表明,线粒体CO I基因作为DNA条形码可以实现仔稚鱼的物种鉴定,且较多能鉴定到种的水平.     

长臂虾科线粒体基因组结构与系统进化分析

综合分析了长臂虾科(Palaemonidae)14 个物种线粒体基因组的全序列, 发现长臂虾科线粒体基因组的长度为 15396~15967 bp, A+T 含量为 58.97%~69.09%; 长臂虾科内不同属线粒体基因组的基因排列各不相同, 与十足目 (Decapoda)线粒体基因组的原始排列相比, 除沼虾属(Macrobrachium)的基因排列顺序未发生改变外, 长臂虾属 (Palaemon)、叶颚虾属(Hymenocera)和贝隐虾属(Anchistus)的基因排列顺序均发生了不同程度的变化, 此外贝隐虾属(Anchistus)还出现基因缺失现象; 长臂虾属和沼虾属线粒体 13 个蛋白编码基因(protein-coding genes, PCGs)的 Ka/Ks<<1, 显示出很强的纯化选择; 长臂虾科 13 PCGs 在密码子的使用中, 被编码的氨基酸均体现相似的偏好性; 在线粒体基因组的差异位点分析中, 发现 nd5、nd4 和 nd2 基因为理想的分子标记。基于 13 PCGs 系统发育结果表明, 长臂虾属与沼虾属互为姊妹关系, 叶颚虾属和贝隐虾属单独聚为一大支。分子钟的估算结果推测长臂虾科物种可能起源于二叠纪, 随后进一步分化为具有现代表征的长臂虾种类。本研究为快速鉴定长臂虾科生物提供了可靠的分子标记, 并为分析长臂虾科物种遗传多样性提供理论基础。     

Diversity and functional importance of coral-feeding fishes on tropical coral reefs

Fishes that feed from live corals (corallivores) are a conspicuous component of healthy coral reef environments. However, knowledge of the occurrence and ecological significance of this feeding mode is fragmentary. Historically, very few fish were considered capable of feeding from live coral, and those few that did were considered ecologically insignificant. More recently, the role of corallivores has been re-evaluated; published records document 128 corallivorous fish species from 11 different families, with 69 of these belonging to the family Chaetodontidae. Other families, including the Labridae, Tetraodontidae, Balistidae, Monacanthidae, Pomacentridae and Scaridae, all have between seven and ten coral-feeding species. One-third of coral-feeding fishes feed almost exclusively on corals, with more than 80% of their diet based on coral. Corallivorous fish show distinct prey preferences and consume only a small subset of available corals, usually the genera Acropora , Pocillopora and Porites . This selective predation by corallivores can limit abundance and distribution of preferred corals. Chronic predation by corallivores may also exacerbate effects of coral disturbance (e.g. climate-induced coral bleaching), impeding reef recovery and causing further coral loss. Conversely, the cover of preferred corals can be a primary determinant of corallivore abundance and physiological condition. Owing to this close association, obligate corallivores invariably decline in response to loss of coral cover. Increased knowledge of the number of corallivores and their diets suggest that this feeding mode is more important to coral reef food webs than traditionally thought.     

Spatial and temporal diversity of fish captured by light traps in various habitats in coastal waters in the oldest marine protected area in Vietnam and its implications for conservation

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分析了中国鯻科鱼类4属6种17尾鱼的线粒体16S rRNA基因3′端部分序列特征。结果表明,在鯻科鱼类507 bp的碱基序列中有变异位点57个,简约信息位点55个,A+T含量(50.3%)高于G+C含量(49.8%),转换/颠换比为2.7。在邻接树和最大似然树上,鯻科鱼类聚类为2个分支,其中鯻属细鳞鯻独立为一支,其余属种组成另一分支,不支持Vari(1978)关于鯻科15个属中匀鯻属为最早分化类群的结论;鯻属的细鳞鯻和鯻鱼并未聚类在一起,表明二者亲缘关系较远,或为不同属,与Lee等报道相一致。由于鯻科鱼类属种较多,本文所分析的样品有限,且鯻科系统分类长期存在争议,因而需要采用多个线粒体和核基因联合分析更多属种,才能确定可否另立新属,并进一步明确鯻科鱼类的系统发育关系。     

团头鲂3个地理种群杂交效果的配合力和微卫星标记预测

根据南沙群岛西南部陆架海域鱼类调查研究的历史资料, 采用Nelson分类系统对鱼类物种组成进行了系统整理, 并从不同的分类阶元上分析了南沙群岛西南部陆架海域鱼类的多样性。分类阶元包含指数(TINCLi), 分类多样性指数(△+和∧+), 目级和科级水平丰富度(Rr)以及目级和科级水平相似性系数(C)为本研究的主要研究指数。结果显示, 南沙群岛西南部陆架海域共记录鱼类20目110科206属, 以鲈形目(Perciformes)种类居绝对优势。采用分类阶元包含指数分析, 南沙群岛西南部陆架海域鱼类组成目、科、属所分别拥有的(科、属、种), (属、种)和(种)的平均数目分别为(5.5、10.30、14.85), (1.87、2.70)和(1.44); 在目级水平上, 比高纬度的东海陆架、大亚湾和东沙群岛海域高, 并且与纬度梯度成反比, 证明低纬度海域的鱼类分类多样性低, 种类组成主要集中分布于较少数的分类类群。使用PRIMER5.2软件计算了平均分类差异指数△+和分类差异变异指数∧+, 数值分别为65.8和110.0, 其中△+接近于东海陆架(65.7), 高于东沙群岛(55.2)和大亚湾海域(62.2), 说明南沙群岛西南部陆架海域鱼类的亲缘关系接近于东海陆架海域鱼类, 比东沙群岛和大亚湾海域鱼类的远。与三个海区的相似性系数比较结果显示, 在目级和科级水平上, 南沙群岛西南部陆架海域鱼类组成同大亚湾海域更为接近。     

Using genetic methods for analysis of fish meals and feeds employed in Greek mariculture

Large quantities of high protein fish meals are needed to sustain cultured species and thus the impact to marine ecosystem has been highly discussed. The aim of this study was to apply a PCR‐cloning methodology for a robust insight into the composition of commercial fish meals and feeds for farmed species of the Greek mariculture, assessing the risk posed by aquaculture to marine ecosystems but also the risk posed by commercial fish feeds to the increase in trophic level of species farmed in Greece. 89% of the sequences were identified to species level and only 11% to genus/family level. Overall, a total of 49 taxa were identified (44 fish species/taxon, five non‐fish species/taxon). Even though small pelagic fish like Engraulis sp. were the main portion, a wide range of species constituted the fish meals and feeds. Plant and animal species were also detected as an alternative protein source. Feed products employed in Greek mariculture still contain large portions of fish meals which increase the mean trophic level of farmed species causing a farming up trend. The results emphasize that such molecular methodologies are needed to certify aquafeeds allowing fish feed producers to demonstrate their commitment to sustainable aquaculture.     

Identification of imported Chirimen at the genus level by a direct sequencing method using mitochondrial partial 16S rDNA region

ABSTRACT:   Quality control of imported fish products is a constant challenge for Customs officials. To help officials access the quality and integrity of imported fish products, and to enhance the effectiveness and accuracy of product screening, a method was developed to identify boiled and dried larval and juvenile sardines (Chirimen) at the genus level by direct sequencing using a partial sequence (606 bp) of mitochondrial 16S rDNA. A phylogenetic tree was constructed, based on nucleotide sequences of seven kinds of Chirimen imported from four countries with 16 standard samples belonging to the orders Clupeiformes and Salmoniformes. To identify the fish genera in imported Chirimen, bootstrap values and branch lengths of the phylogenetic tree as well as genetic distances were used. The data revealed that Chirimen imported from China and Korea contains species of the genus Engraulis . Regarding other imported Chirimen, the least that could be verified was that the sardine ingredients do not belong to regulated genera.     

Climate change and the future for coral reef fishes

Climate change will impact coral-reef fishes through effects on individual performance, trophic linkages, recruitment dynamics, population connectivity and other ecosystem processes. The most immediate impacts will be a loss of diversity and changes to fish community composition as a result of coral bleaching. Coral-dependent fishes suffer the most rapid population declines as coral is lost; however, many other species will exhibit long-term declines due to loss of settlement habitat and erosion of habitat structural complexity. Increased ocean temperature will affect the physiological performance and behaviour of coral reef fishes, especially during their early life history. Small temperature increases might favour larval development, but this could be counteracted by negative effects on adult reproduction. Already variable recruitment will become even more unpredictable. This will make optimal harvest strategies for coral reef fisheries more difficult to determine and populations more susceptible to overfishing. A substantial number of species could exhibit range shifts, with implications for extinction risk of small-range species near the margins of reef development. There are critical gaps in our knowledge of how climate change will affect tropical marine fishes. Predictions are often based on temperate examples, which may be inappropriate for tropical species. Improved projections of how ocean currents and primary productivity will change are needed to better predict how reef fish population dynamics and connectivity patterns will change. Finally, the potential for adaptation to climate change needs more attention. Many coral reef fishes have geographical ranges spanning a wide temperature gradient and some have short generation times. These characteristics are conducive to acclimation or local adaptation to climate change and provide hope that the more resilient species will persist if immediate action is taken to stabilize Earth's climate.     

Profile of the Marine Aquarium Fish Trade in Hong Kong

A market survey and review of government statistics were carried out to establish imports and exports of marine ornamental fishes into and out of Hong Kong, and to examine the local trade in terms of volume, value and species composition. Official government import figures for marine aquarium fishes were available from 1984 to 1991 and from 1997 and 1998. When compared with net imports, export and re-export volumes over the 15-year period were small, indicating that most imports entered the local market, or were exported unrecorded. The market survey of marine aquarium shops in Hong Kong was carried out between August 1996 and January 1997. From this survey, an annual estimate of 957,563 coral reef fish was calculated for the local trade, valued at HK$57,453,780, with a mean retail price of HK$60 per fish. These figures account for an estimated 2–3% of the global value and volume of marine aquarium fish trade, according to 1992 figures and, compared with government figures, indicate that official declarations of imports are underreported by at least 2–3 fold. A total of 342 marine aquarium fish species, from 49 families, were recorded with about 60% belonging to the families Labridae, Chaetodontidae, Pomacanthidae and Pomacentridae. It was estimated that a large proportion of the trade was in juvenile fishes and almost all fish were less than 10cm in standard length. It was common to see fishes in poor condition, species hard to maintain in captivity, or those listed for conservation concern, on sale. There are no regulations for local traders of live marine fishes other than those relating to either protected species or animal welfare. Licensing of traders in marine fishes could be introduced under existing Hong Kong legislation if fishes were to be re-classified as animals. This is strongly recommended as a step towards regulating and managing the trade in Hong Kong.     

Phylogenetic relationships in grey mullets (Mugilidae) in a Tunisian lagoon

The Mugilidae family is an important fish group representing a major source for fisheries and aquaculture. In the south Mediterranean bank, no data are available on this fauna, except for some morphological studies on Tunisian samples. In this study, 16 allozymic loci were used to investigate the phylogenetic relationships within Tunisian mugilids. The results obtained from Hergla lagoon samples highlight five operational taxonomic unit corresponding to the well‐known species (Liza aurata, Liza ramada, Liza saliens, Chelon labrosus, Mugil cephalus). Several loci appeared to be diagnostic of these species, but in contrast to Greek mugilid samples, we did not find any diagnostic locus fixed differently for the five species. These results can help aquaculture units to identify accurately the mullet species they subsequently use for stocking aquaculture ponds and inland waters. However, species identity represents very important information, as each species has a different growth rate and salinity tolerance. On the other hand, when compared with North Mediterranean Mugilidae analysed until now, Tunisian samples show a genetic differentiation that could be related to different physicochemical conditions between the North and South banks, similar to those recorded in the eastern and western two Mediterranean basins separated by the Siculo‐Tunisian strait. In addition, this study confirms the morphological taxonomy, except for the subdivision of the Liza genus into two sub‐genera. The phylogenetic tree is in agreement with that on Languedoc Mugilidae samples (France), indicating that the subdivision of the Liza genus into two sub‐genera appears to be without any genetic base.