广东沿海洛氏角毛藻复合群物种多样性的探究

为了澄清洛氏角毛藻复合群的物种多样性, 并明确洛氏角毛藻和并基角毛藻的种间界限, 文章以广东沿海为例, 建立了洛氏角毛藻复合群的22个单克隆培养株系, 利用光学显微镜和电子显微镜技术, 开展了基于生活史的连续形态学观察; 结合基于核糖体大亚基编码基因D1-D3区序列的分子系统学分析。结果表明: 文章支持近年研究的观点, 认为“角毛基部并行融合”不是稳定特征, 不宜继续作为并基角毛藻的标志特征; 发现洛氏角毛藻形态相似藻株聚类在2个分支上, 确认了洛氏角毛藻确实存在隐藏的物种多样性。其中一类与目前认知的洛氏角毛藻最为接近, 而另一类在角毛孔纹、休眠孢子上存在明显区别。经过对洛氏角毛藻历史文献的对比研究, 发现洛氏角毛藻的物种界定存在混乱和模糊的情况, 如休眠孢子形态, 及其初生壳面是否具有二叉状分布结构等, 均存在相互矛盾或不一致的历史报道。     

短孢角毛藻相似种类的形态学和系统学研究

短孢角毛藻的形态特征多变,容易导致物种的错误鉴定,并且其与若干相似种类之间存在物种混淆的情况。基于建立的单克隆培养藻株,结合生活史中形态特征的连续观察,以及基于目标基因序列分析的分子系统学数据,对短孢角毛藻(Chaetoceros brevis Schütt)及其相似种类的形态学和系统学进行了研究。丰富了短孢角毛藻休眠孢子的形态学描述,报道了一个新的特征:休眠孢子壳面上刺的末端具有帽状结构,该特征未报道于其他角毛藻种类,可作为短孢角毛藻的标志性特征。报道了我国角毛藻属的1个新记录种——拟短孢角毛藻(C.pseudobrevis Pavillard),该种具有独特的形态学特征,以及独立的系统学位置,应是一个独立的物种,而非短孢角毛藻的同种异名。垂缘组角毛藻的种类在系统发育树中显示出较好的单系起源,链端角毛及其走势是否可以作为该类群的共享特征,目前尚难做出定论。     

异角毛藻和平滑角毛藻的分类学修订

为了澄清异角毛藻和平滑角毛藻的分类学疑问,在广东南澳岛海域建立了单克隆培养株系,利用光学显微镜和电子显微镜技术,开展了基于生活史的连续形态学观察,发现粗大角毛的延伸方向易变,是不稳定特征,不宜继续赋予分类学价值。对核糖体大亚基编码基因的D1～D3区序列进行扩增和分析,结果异角毛藻和平滑角毛藻的目标基因序列基本一致,仅有1个平滑角毛藻株系存在2个差异碱基。因此,两者具有一致的系统学位置,属于同一物种,平滑角毛藻应该是异角毛藻的同种异名。     

中国沿海洛氏角毛藻复合群的多样性组成及地理分布

洛氏角毛藻复合群(Chaetoceros lorenzianus complex)指具有与洛氏角毛藻相似形态学特征的物种集合, 它们广泛分布于全球近岸水域。近年国际上关于该复合群的分类学研究取得新进展, 而我国相关研究仍较为滞后。为了弄清我国沿海洛氏角毛藻复合群的物种多样性, 明确物种信息, 厘清种间界限, 为相关研究提供准确的物种鉴定依据, 本研究陆续在中国沿海建立了该复合群的332个单克隆培养株系, 利用光学显微镜、扫描电镜和透射电镜进行了较为详尽的形态学研究, 基于核糖体大亚基编码基因D1-D3区序列, 构建了分子系统学关系。结果表明其形态聚类与分子系统学结论相一致, 显示我国洛氏角毛藻复合群具有较高的物种多样性, 共鉴定到5个物种, 分别是并基角毛藻(C. decipiens)、优美角毛藻(C. elegans)、平孢角毛藻(C. laevisporus)、曼纳角毛藻(C. mannaii)和稀树角毛藻(C. pauciramosus)。研究表明传统认知的光镜下特征, 如群体特征、角毛走势等易变化, 其分类学价值需谨慎应用。角毛的超微结构, 如角毛孔纹的形状、大小、密度等是有效的种间区别特征, 休眠孢子亦是重要的物种识别依据。并基角毛藻和平孢角毛藻在我国沿岸的分布范围最为广泛, 而稀树角毛藻的分布较为有限。     

CO_2浓度倍增对牟氏角毛藻生长和光合作用的影响

高浓度（５％)ＣＯ２对微藻光合作用特征和固碳机制的影响已有了广泛的研究,但ＣＯ２提高数倍对微藻的影响报道还不多。本文以牟氏角毛藻为材料,研究其在ＣＯ２浓度倍增条件下（７００ｕＬＬ－１）的生长和光合作用特点,以期了解ＣＯ２浓度的倍增影响微藻类的生理生化效应和机制。另外,牟氏角毛藻是常用的饵料藻种,含有丰富的多不饱和脂肪酸[１］,所以还有一定的实践意义。１ 材料与方法１．１ 材料与培养 牟氏角毛藻（ＣｈａｅｔｏｃｅｒｏｓｍｕｅｌｌｅｒｉｍｕｅｌｅｒｉＬｅｍｍ．）由中国科学院海洋研究所提供。对数生长期的藻…     

营养强化时褶皱臂尾轮虫对饵料微藻的摄食

选取适宜浓度利用单种微藻和混合微藻对轮虫进行营养强化,采用实验生态学方法研究了轮虫滤水率和摄食率的动态变化.结果表明:微藻浓度、微藻种类和培养时间均对轮虫的滤水率和摄食率有显著影响;轮虫对几种单种微藻的滤水率和摄食率均随培养时间的延长而下降,在实验条件下,6h内轮虫对3种微藻的滤水率大小顺序为小球藻＞球等鞭金藻＞牟氏角毛藻,12h内轮虫对3种和,微藻的滤水率大小顺序则为球等鞭金藻＞小球藻＞牟氏角毛藻;轮虫在混合微藻中的选择顺序为球等鞭金藻＞小球藻＞牟氏角毛藻.     

大亚湾水域并基拟菱形藻的种类鉴定和产毒特征分析

为了丰富我国海域拟菱形藻(Pseudo-nitzschia Peragallo)的物种多样性, 并澄清其产毒特征, 研究从广东大亚湾海域分离并建立了一株拟菱形藻的单克隆培养株系MC298, 通过光学显微镜下的群体特征和透射电镜下的超微形态特征观察, 结合基于核糖体转录间隔区(Internal Transcribed Spacer, ITS)的分子系统学数据, 以及基于ITS2转录RNA的二级结构分析, 鉴定到我国拟菱形藻属的1个新记录种: 并基拟菱形藻P. decipiens Lundholm & Moestrup。研究对其形态学特征进行了较为详细地描述, 并与相似种进行了比较, 还对其ITS2-RNA的特有标志结构进行了阐述。同时, 利用高效液相色谱-质谱联用法(Liquid chromatography tandem mass spectrometry, LC-MS/MS)对该藻株的产毒特征进行了检测, 结果未检测到DA的存在。研究不仅丰富了我国拟菱形藻属的物种多样性, 也可为拟菱形藻的产毒特征研究提供基础数据。     

我国3个新记录海链藻种类的形态学研究

利用光镜(LM)和扫描电镜(SEM)技术,对采自我国沿海海域的自然水样,以及分离获得的单克隆培养藻株进行了形态学观察,针对其中的海链藻属Thalassiosira Cleve种类开展了形态分类学的专题研究。报道了3个新记录种类,分别是安达曼海链藻Thalassiosira andamanica Gedde、缢缩海链藻Thalassiosira con-stricta Gaarder和双线海链藻Thalassiosira duostra Pienaar。对每个种类的形态学特征、生活习性和生态分布进行了描述,提供了光镜和电镜照片,并对相似种类的形态学特征进行了比较研究。     

我国沿海闭合突海链藻的形态学研究

闭合突是一类特殊的突起, 仅报道于少数海链藻种类, 其形态学特征和排列方式是重要的分类学依据。我国关于闭合突海链藻的报道较少。研究利用扫描电镜(SEM)技术, 对采自我国沿海海域的自然水样, 以及分离获得的单克隆培养藻株进行了形态学观察, 针对其中的海链藻属Thalassiosira Cleve种类开展了形态分类学的专题研究。报道了5个具有闭合突的海链藻种类, 分别是西达礁海链藻T. cedarkeyensis Prasad、偏心海链藻T. eccentrica (Ehrenberg) Cleve、伦德海链藻T. lundiana Fryxell、细孔海链藻T. punctigera (Castracane) Hasle和管状海链藻T. tubifera Fryxell, 其中有3个为我国新记录种类: 西达礁海链藻、细孔海链藻和管状海链藻。对每个种类的形态学特征和生境进行了描述, 提供了扫描电镜照片, 对相似种类的形态学特征进行了比较研究, 分析了突起特征的变化类型及其分类学价值。       

我国沿海线形亚属海链藻的形态学研究

壳面孔纹的形态学特征是海链藻的重要分类学依据之一。线形亚属的海链藻种类大多是从圆筛藻属修订而来, 它们之间的区别特征细微, 需电镜下观察才能准确鉴定。我国关于线形亚属海链藻的报道较少。研究利用电镜(EM)技术, 对采自我国沿海海域的自然水样, 以及分离获得的单克隆培养藻株进行了形态学观察, 针对其中的海链藻属Thalassiosira Cleve种类开展了形态分类学的专题研究。报道了6个隶属于线形亚属的海链藻种类, 分别是紧密海链藻T. densannula Hasle &; Fryxell、微小海链藻T. exigua Fryxell &; Hasle、线形海链藻T. lineata Jousé、微线形海链藻T. nanolineata (Mann) Fryxell &; Hasle、结线形海链藻T. nodulolineata (Hendey) Hasle &; Fryxell和柔弱海链藻T. tenera Proschkina-Lavrenko, 其中有4个为我国新记录种类: 紧密海链藻、线形海链藻、微线形海链藻和结线形海链藻。对每个种类的形态学特征、生活习性和生态分布进行了描述, 提供了电镜照片。对相似种类的形态学特征进行了比较研究, 分析了孔纹特征的变化类型及其分类学意义。     

New investigation technique for thoracic sclerites of mosquitoes (Diptera, Culicidae) for the correct identification of genera and species

A new technique for studying the external morphology of thoracic sclerites of mosquitoes (Diptera: Culicidae) was developed. According to this method, the shape of sclerites and the position of setae and scales can be examined using SEM or light microscopy even in cases when setae or scales have been lost. The method can be recommended for the damaged material which is often obtained during sampling. The bases of setae usually exceed 100 μm in diameter and thus differ significantly from the bases of scales which are about 30 μm in diameter. Analysis of the structure of sclerites and the arrangement of setae and scales on them in different specimens of one species may reveal intraspecific variation of this character complex. Comparison of species from different genera of the family Culicidae, as well as comparison of closely related species, will probably help to estimate the diagnostic value of thoracic sclerites as morphological characters.     

Evidence of hidden biodiversity, ongoing speciation and diverse patterns of genetic structure in giant Antarctic amphipods

Recent molecular research on Antarctic benthic organisms has challenged traditional taxonomic classifications, suggesting that our current perceptions of Antarctic biodiversity and species distributions must be thoroughly revised. Furthermore, genetic differentiation at the intraspecific level remains poorly understood, particularly in eastern Antarctica. We addressed these issues using DNA sequence data for two sibling amphipod species that could be collected on a circum-Antarctic scale: Eusirus perdentatus and Eusirus giganteus. Haplotype networks and Bayesian phylogenies based on mitochondrial (COI, CytB) and nuclear (ITS2) DNA provided strong evidence of multiple cryptic species of Eusirus, with several occurring in sympatry and at least one likely to have a true circum-Antarctic distribution. Within species, gene flow was often highly restricted, consistent with a brooding life history and in some cases suggestive of current or future allopatric speciation. Patterns of genetic structure were not always predictable: one cryptic species showed preliminary evidence of high genetic differentiation across ～150 km in eastern Antarctica (F(ST) > 0.47, P < 0.01), yet another was remarkably homogenous across ～5000 km (F(ST) = 0.00, P = 1.00). Genetic diversity also varied among cryptic species, independent of sample size (π = 0.00-0.99). These results indicate several hidden levels of genetic complexity in these Antarctic amphipods that are neither apparent from previous taxonomic or ecological studies nor predictable from their life history. Such genetic diversity and structure may reflect different modes of survival for Antarctic benthic organisms during historic glacial cycles, and/or subsequent re-establishment of populations on the shelf, and highlight our misunderstanding of Antarctic marine species diversity.     

Taxonomic revision of the Mediterranean Plakina Schulze (Porifera, Demospongiae, Homoscleromorpha)

Only three species of the sponge genus Plakina Schulze have been described from the Mediterranean since 1880, in spite of a large amount of allegedly intraspecific variability in morphological characters. However, recent genetic studies based on electrophoretic techniques have revealed extensive cryptic speciation in north-western Mediterranean Plakina , demonstrating that most of this variation was interspecific rather than intraspecific. We describe in detail the morphology and anatomy of four new species of Plakina from the Mediterranean– P. crypta, P. weinbergi, P. endoumensis and P. jani –of which the latter two were discovered through allozyme electrophoresis. Plakina monolopha Schulze and P. trilopha Schulze are redescribed, and their morphological and geographical limits are discussed along with those of P. dilopha Schulze. Accurate analysis of the internal anatomy and of the shape and ramification pattern of lophose spicules in scanning electron microscopy provides new, powerful morphological criteria for species discrimination in Plakina. More widespread use of such new taxonomic characters should provide evidence against the alleged cosmopolitanism of some Plakina species, thus generating an increase in estimates of the biodiversity of plakinids.     

The dark side of pseudoscorpion diversity: The German Barcode of Life campaign reveals high levels of undocumented diversity in European false scorpions

DNA barcoding is particularly useful for identification and species delimitation in taxa with conserved morphology. Pseudoscorpions are arachnids with high prevalence of morphological crypsis. Here, we present the first comprehensive DNA barcode library for Central European Pseudoscorpiones, covering 70% of the German pseudoscorpion fauna (35 out of 50 species). For 21 species, we provide the first publicly available COI barcodes, including the rare Anthrenochernes stellae Lohmander, a species protected by the FFH Habitats Directive. The pattern of intraspecific COI variation and interspecific COI variation (i.e., presence of a barcode gap) generally allows application of the DNA barcoding approach, but revision of current taxonomic designations is indicated in several taxa. Sequences of 36 morphospecies were assigned to 74 BINs (barcode index numbers). This unusually high number of intraspecific BINs can be explained by the presence of overlooked cryptic species and by the accelerated substitution rate in the mitochondrial genome of pseudoscorpions, as known from previous studies. Therefore, BINs may not be an appropriate proxy for species numbers in pseudoscorpions, while partitions built with the ASAP algorithm (Assemble Species by Automatic Partitioning) correspond well with putative species. ASAP delineated 51 taxonomic units from our data, an increase of 42% compared with the present taxonomy. The Neobisium carcionoides complex, currently considered a polymorphic species, represents an outstanding example of cryptic diversity: 154 sequences from our dataset were allocated to 23 BINs and 12 ASAP units.     

Coalescent Method in Conjunction with Niche Modeling Reveals Cryptic Diversity among Centipedes in the Western Ghats of South India

Background

There has been growing interest in integrative taxonomy that uses data from multiple disciplines for species delimitation. Typically, in such studies, monophyly is taken as a proxy for taxonomic distinctiveness and these units are treated as potential species. However, monophyly could arise due to stochastic processes. Thus here, we have employed a recently developed tool based on coalescent approach to ascertain the taxonomic distinctiveness of various monophyletic units. Subsequently, the species status of these taxonomic units was further tested using corroborative evidence from morphology and ecology. This inter-disciplinary approach was implemented on endemic centipedes of the genus Digitipes (Attems 1930) from the Western Ghats (WG) biodiversity hotspot of India. The species of the genus Digitipes are morphologically conserved, despite their ancient late Cretaceous origin.

Principal Findings

Our coalescent analysis based on mitochondrial dataset indicated the presence of nine putative species. The integrative approach, which includes nuclear, morphology, and climate datasets supported distinctiveness of eight putative species, of which three represent described species and five were new species. Among the five new species, three were morphologically cryptic species, emphasizing the effectiveness of this approach in discovering cryptic diversity in less explored areas of the tropics like the WG. In addition, species pairs showed variable divergence along the molecular, morphological and climate axes.

Conclusions

A multidisciplinary approach illustrated here is successful in discovering cryptic diversity with an indication that the current estimates of invertebrate species richness for the WG might have been underestimated. Additionally, the importance of measuring multiple secondary properties of species while defining species boundaries was highlighted given variable divergence of each species pair across the disciplines.     

Systematics of malaria vectors with particular reference to the Anopheles punctulatus group

The appearance of groups and complexes of closely related cryptic or sibling species in many of the anopheline taxa has impeded studies on malaria transmission and the evaluation of control strategies which have relied on morphological characters to identify the vector species involved. The advantages of morphological identification are low cost, speed and simplicity, which allow large numbers of specimens to be processed rapidly in the field. The need for accurate identification is crucial, as time and money may be wasted in studying and controlling species of no medical importance. Various techniques such as cross-mating, chromosome studies and allozyme analysis have been developed to resolve problems of identifying sibling species, though none, as yet, can match the speed and simplicity afforded by morphology markers. The latest of these identification methods comes from advances that have been made in DNA-based technology. Although costly and requiring fairly sophisticated laboratory support, methods such as DNA probe hybridisation and PCR are the quickest and most user-friendly to date. The use of DNA has other advantages in the study of intraspecific differences and in providing characters for phylogenetic studies. This review looks at the development of DNA-based techniques for taxonomic and systematic studies of anopheline mosquitoes. The Anopheles punctulatus group of the southwest Pacific is featured as an example of how this technology has been applied and how it has progressed.