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。     

并基角毛藻若干分类学疑问的初步探讨

并基角毛藻是硅藻门角毛藻属的典型种类, 以“相邻角毛基部并行融合”作为标志性特征, 但该特征是否稳定, 及如何赋予其分类学价值?尚存较多争议。研究以符合现存并基角毛藻分类标准的藻株作为目标藻株, 采用毛细管显微操作技术从我国沿海代表水域分离藻株, 建立了目标藻株的单克隆培养株系, 利用光学显微镜、扫描电镜和透射电镜分别对其形态学特征进行观察, 同时还扩增了核糖体大亚基编码基因的D1-D3区序列, 并构建系统进化树。综合分析形态学特征和分子系统学数据, 初步得到以下结论: (1)具有相同遗传特征的并基角毛藻株系在“相邻角毛基部并行融合”特征上具有不稳定性, 即“相邻角毛基部并行融合”不能作为并基角毛藻种类的标志性特征; (2)并基角毛藻和洛氏角毛藻作为相似种类, 均显示出较高的物种多样性, 高于目前的认知, 预示着或许存在隐形种或拟隐形种; (3)推测并基角毛藻单胞变型只是并基角毛藻原种的一个生活史阶段, 是并基角毛藻的同种异名, 不宜继续保留其独立的分类学地位。     

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。     

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。     

氮、硅限制对赤潮藻扁面角毛藻休眠孢子形成的影响

休眠孢子的形成对于赤潮藻种群的保存、延续以及分布扩散等均具有重要的意义。通过单因子营养限制研究氮、硅对赤潮藻扁面角毛藻（Chaetoceros compressus）休眠孢子形成的影响,结果表明：培养基中氮的初始浓度对休眠孢子的出现时间有一定影响。氮的初始浓度越低,休眠孢子出现的时间越早：反之,氮的初始浓度越高,休眠孢子出现的时间越晚。氮缺乏是硅藻形成休眠孢子的必需条件之一,当培养基中氮含量低于10μmol·L^-1时,扁面角毛藻可以形成休眠孢子。氮缺乏诱发的休眠孢子的形成需要大量的硅,当培养基中硅含量低于23μmol·L^-1时,即使氮缺乏,扁面角毛藻也几乎不再继续形成休眠孢子。这说明硅藻休眠孢子的形成不仅受氮浓度的影响,还与硅浓度有关。     

氮、硅限制对赤潮藻扁面角毛藻休眠孢子形成的影响

休眠孢子的形成对于赤潮藻种群的保存、延续以及分布扩散等均具有重要的意义。通过单因子营养限制研究氮、硅对赤潮藻扁面角毛藻 (Chaetoceros compressus )休眠孢子形成的影响, 结果表明: 培养基中氮的初始浓度对休眠孢子的出现时间有一定影响。氮的初始浓度越低, 休眠孢子出现的时间越早; 反之, 氮的初始浓度越高, 休眠孢子出现的时间越晚。氮缺乏是硅藻形成休眠孢子的必需条件之一, 当培养基中氮含量低于10 mmol.L^-1时, 扁面角毛藻可以形成休眠孢子。氮缺乏诱发的休眠孢子的形成需要大量的硅, 当培养基中硅含量低于23 mmol.L^-1时, 即使氮缺乏, 扁面角毛藻也几乎不再继续形成休眠孢子。这说明硅藻休眠孢子的形成不仅受氮浓度的影响, 还与硅浓度有关。     

大亚湾角毛藻细胞数量波动及其与环境因子关系的多元分析

研究了1997年6月～1998年7月及1999和2000年春季大亚湾角毛藻（Chaetoceros spp.）细胞数量变动及其与环境因子之间的关系,并进行了多因变量方差分析、多元线性逐步回归分析及主成分分析.角毛藻是大亚湾浮游植物常见优势属,优势物种主要为窄隙角毛藻（C.affinis）、柔弱角毛藻（C.debilis）、远距角毛藻（C.distans）和劳氏角毛藻（C.lorenzianus）等.1997～1998年度角毛藻细胞数量出现了明显的春季高峰,最高细胞数量和百分比含量分别达6680.6 cells/ml和99.78%,而且在其他季节也出现了数量不等的峰值,DIN的消耗在一定程度上导致了角毛藻水华的消退.多元分析结果表明,水温和营养元素之间的比例是影响角毛藻细胞数量变动的主要因子,此外DIN和盐度对角毛藻细胞数量变动也产生较明显的影响.     

粤东柘林湾角毛藻（Chaetoceros）群落生态学特性的季节变化

于2004年5月至2005年2月对粤东大规模养殖区柘林湾的角毛藻(Chaetoceros)群落的时空分布进行了季节性调查.调查期间,共发现角毛藻54种(含变种、变型),包括3个未知角毛藻种,其中柔弱角毛藻(C.debilis),双孢角毛藻(C didymus vat.didymus),劳氏角毛藻(C.lorenzianus),小角毛藻(C.minutissimus)和聚生角毛藻(Chaetoceros socialis)均为柘林湾全年出现种.柘林湾角毛藻种类丰富度和细胞丰度的季节变化均呈现高温季节高低温季节低的特点,而平面分布均表现出由湾内向湾外递增的趋势.整体而言,是一个角毛藻种类数很高而细胞丰度较低的海湾.分析表明,水温是影响角毛藻种类丰富度和细胞丰度季节变化的关键因素,而光照、种间竞争和浮游动物群落结构的组成是影响其时空分布的主要因素.     

拟荠角孢属一新种——云南拟荠角孢

从云南省采集的飞机草菌绒孢[Mycovellosiella eupatorii-Odorati(Yen)Yen]一批标本中,我们发现其中一些常混有主要生于飞机草(Eupatorium odoratum L.)叶毛上的另一种菌。经研究其形态特征,确定是拟荠角孢属(Alysidiopsis)一新种,云南拟荠角孢(Alysidiopsisyunnanensis sp.nov.)。本文对此新种作了拉丁文和汉文描述并附形态图予以报道。     

链格孢菌酯酶同工酶分析及其分类学意义

分析了20株链格孢菌的酯酶同工酶聚丙烯酰胺凝胶电泳图谱,并将酶谱进行聚类分析。结果发现:20株链格孢菌在37%的相似系数处明显分为大孢子组和小孢子组;大孢子组在52%的相似系数处分为6组,每组代表1个种,小孢子组在较高的相似性水平上分组与形态分组结果基本一致。本试验结果还表明:酯酶同工酶电泳方法简单易行,能准确反映种间的微小差异,适用于链格孢属真菌的种级分类,可作为传统形态学分类的一个重要辅助手段。     

Metabolic fingerprinting reveals differences between northern and southern strains of the cryptic diatom Chaetoceros socialis

Morphology and molecular phylogeny constitute the structural elements of diatom taxonomy. These approaches do not, however, give information on the functioning of taxa. Additional methods to serve a more integrated and wide-ranging taxonomy have therefore been called for. Metabolic fingerprinting is one approach used within the field of metabolomics, often applied in classification of samples. Here we apply metabolic fingerprinting in a taxonomic study of a cryptic diatom species. Strains of the cosmopolitan diatom Chaetoceros socialis from two geographical areas; the north-east Atlantic and Arctic and the Gulf of Naples, were cultivated at three different temperatures; 2.5, 8 and 13°C. The strains from the two different geographical areas exhibited different growth rates as well as different photosynthetic efficiencies. Algal extracts, collected at the end of the growth experiments, were analysed by Ultra-Performance Liquid Chromatography High Resolution Mass Spectrometry. The two groups of strains were separated by principal component analysis of their metabolic fingerprints. Analysis of the data revealed both qualitative and quantitative differences in metabolite markers. These phenotypic differences reinforce differences also found for morphology, phylogenetic markers and growth rates, and point at different adaptive characteristics in organisms living under different temperature regimes.     

Two new species in the Chaetoceros socialis complex (Bacillariophyta): C. sporotruncatus and C. dichatoensis,and characterization of its relatives,C. radicans and C. cinctus

The diatom genus Chaetoceros is one of the most abundant and diverse phytoplankton in marine and brackish waters worldwide. Within this genus, Chaetoceros socialis has been cited as one of the most common species. However, recent studies from different geographic areas have shown the presence of pseudo‐cryptic diversity within the C. socialis complex. Members of this complex are characterized by curved chains (primary colonies) aggregating into globular clusters, where one of the four setae of each cell curves toward the center of the cluster and the other three orient outwards. New light and electron microscopy observations as well as molecular data on marine planktonic diatoms from the coastal waters off Chile revealed the presence of two new species, Chaetoceros sporotruncatus sp. nov. and C. dichatoensis. sp. nov. belonging to the C. socialis complex. The two new species are similar to other members of the complex (i.e., C. socialis and C. gelidus) in the primary and secondary structure of the colony, the orientation pattern of the setae, and the valve ultrastructure. The only morphological characters that can be used to differentiate the species of this complex are aspects related to resting spore morphology. The two newly described species are closely related to each other and form a sister clade to C. gelidus in molecular phylogenies. We also provide a phylogenetic status along with the morphological characterization of C. radicans and C. cintus, which are genetically related to the C. socialis complex.     

Morphology and ecology of the bloom-forming diatom <Emphasis Type="Italic">Chaetoceros contortus</Emphasis> from Peter the Great Bay,Sea of Japan

The morphology of the marine planktonic diatom Chaetoceros contortus Schütt from the Sea of Japan is described using light and electron microscopy. Because of morphological similarity, C. contortus was previously synonymized with the close species C. compressus Lauder by Gran and Yendo [22]. Abundant development of C. contortus was observed in 1997 in Amursky Bay (Sea of Japan). This species reached a maximum density of up to 1.3 × 10⁶ cells/liter near the water surface in mid May 1997 in the period of water-column stratification.     

Observations on two closely related species, Chaetoceros tetrastichon and C. dadayi (Bacillariophyceae)

Two tropical-subtropical species of the diatom genus Chaetoceros, C. tetrastichon and C. dadayi were studied in light, scanning and transmission electron microscopy, from plankton samples collected in the Gulf of California, off the coasts of Baja California and the Mediterranean Sea. Both species are usually found to be attached to a tintinnid. The basic structure of both species agrees with previous studies of species belonging to the subgenus Phaeoceros , including the finely perforated cell wall of the valves and the presence of one rimoportula in every valve of the cells in the chain, which is located in the centre of the annulus. A clear costae pattern was found in C. tetrastichon , but it is less apparent in C. dadayi. Other differences between the two species are the direction and morphology of the setae, especially the anterior terminal ones, and the number of girdle bands. Questions on the function of the setae in Chaetoceros continue offering matter for speculation.     

True and false gharials: a nuclear gene phylogeny of crocodylia

The phylogeny of Crocodylia offers an unusual twist on the usual molecules versus morphology story. The true gharial (Gavialis gangeticus) and the false gharial (Tomistoma schlegelii), as their common names imply, have appeared in all cladistic morphological analyses as distantly related species, convergent upon a similar morphology. In contrast, all previous molecular studies have shown them to be sister taxa. We present the first phylogenetic study of Crocodylia using a nuclear gene. We cloned and sequenced the c-myc proto-oncogene from Alligator mississippiensis to facilitate primer design and then sequenced an 1,100-base pair fragment that includes both coding and noncoding regions and informative indels for one species in each extant crocodylian genus and six avian outgroups. Phylogenetic analyses using parsimony, maximum likelihood, and Bayesian inference all strongly agreed on the same tree, which is identical to the tree found in previous molecular analyses: Gavialis and Tomistoma are sister taxa and together are the sister group of Crocodylidae. Kishino-Hasegawa tests rejected the morphological tree in favor of the molecular tree. We excluded long-branch attraction and variation in base composition among taxa as explanations for this topology. To explore the causes of discrepancy between molecular and morphological estimates of crocodylian phylogeny, we examined puzzling features of the morphological data using a priori partitions of the data based on anatomical regions and investigated the effects of different coding schemes for two obvious morphological similarities of the two gharials.     

火丝菌科(盘菌目)部分属的系统学研究

本文以核糖体小亚基(SSUrDNA)为分子标记,对28个属60个种的71个序列片段进行序列分析,探讨火丝菌科的属间亲缘关系。研究结果支持广义的火丝菌科概念,表明该科是单起源的,显示5个主要分支。腐生或与植物共生形成菌根、囊盘被表面具有毛状物的15个属构成A分支,该分支中仅部分属之间的关系比较明确;与苔藓植物生长在一起的4个属构成B分支;C分支包括Otidea和Otideopsis两属,后者与Otidea的成员混杂在一起;D分支仅包括Acervus的成员;E分支由Geopyxis,Tarzetta,Paurocotylis等5个属组成。分子系统学的研究结果与形态学分类系统之间存在一定差异,依据形态特征和超微结构建立的亚科和族均未获得支持。