Phylogenetic relationships of the subclass Peritrichia (Oligohymenophorea, Ciliophora) with emphasis on the genus Epistylis, inferred from small subunit rRNA gene sequences

The peritrichs have been recognized as a higher taxon of ciliates since 1968. However, the phylogenetic relationships among them are still unsettled, and their placement within the class Oligohymenophorea has only been supported by the analysis of the small subunit rRNA gene sequence of Opisthonecta henneguyi. DNA was isolated directly from field-sampled species for PCR, and was used to resolve relationships within the genus Epistylis and to confirm the stability of the placement of peritrichs. Small subunit rRNA gene sequences of Epistylis plicatilis, Epistylis urceolata, Epistylis chysemydis, Epistylis hentscheli, Epistylis wenrichi, and Vorticella campanula were sequenced and analyzed using both distance-matrix and maximum-parsimony methods. In phylogenetic trees, the monophyly of both the genus Epistylis and the subclass Peritrichia was strongly supported, while V. campanula clustered with Vorticella microstoma. The topology in which E. plicatilis and E. hentscheli formed a strongly supported sister clade to E. urceolata, E. chrysemydis, and E. wenrichi was consistent with variations in the thickness of the peristomial lip. We concluded that the peristomial area, especially the. peristomial lip, might be the important phylogenetic character within the genus Epistylis.

海洋动植物中不饱和脂肪酸的研究

不饱和脂肪酯是人及动物生长发育的必需物质。本论文分析了海洋动植物脂和脂肪酸的组成，并对不饱和脂肪酸的分离提纯方法进行了研究。一、海洋动植物的中性脂主要包括三酰甘油和胆固醇。动物极性脂主要为磷脂，包括磷脂酰丝馆酸（PS）、磷脂酸（PA）、磷脂酰肌醇（PI）、溶解性磷脂酰乙醇胺（LPE）、神经鞘磷脂（CAEP）、磷脂酰胆碱（PC）、磷脂酰乙醇胺（PE），双磷脂酰甘油（DPG）。其中LPE、DPG为动物所特有的磷脂。植物极性脂包括PI、PC、PE、双半乳糖苷地酰甘油酯（DGDG）、硫代6-脱氧葡萄糖苷二酰甘油酯（SQDG）、单半乳糖苷二酰甘油酯（MGDG）、磷脂酰甘油（PG）及植物脑苷脂（Plant Cerebrosides）。其中MGDG、DGDG、SQDG为植物所特有的三种糖脂。二、海洋动植物含有丰富的不饱和脂肪酸，多聚不饱和脂肪酸（PUFA）含量尤为显著。灰凹贻贝（Crenomythilus graiaynus）的二十碳五烯酸（EPA）含量为10.6%,二十碳四烯酸（AA）为3.7%，二十二碳六烯酸（DHA）为14.26%。中间球海胆（Strongylocentrotus intermedius）的AA含量为7.36%，EPA为15.98%。内枝多管藻（Polysiphonia morroωii)PUFA含量非常高，AA占2.43%，20:4ω3占4.97%，EPA占45.03%。裙带菜（Undaria pinnatifida）的PUFA中，十八碳二烯酸占6.16%，AA占12.99%，EPA占7.83%这些PUFA大都为人及动物生长发育的必需脂肪酸。三、尿素诱导法是大量提取混合PUFA的有效方法。在产品中，饱和脂肪酸的含量可以忽略不计，单烯脂肪酸含量降低很多；高吸附能力的柱层析是提取PUFA单体的较好方法，其产物的纯度达98%以上。

三种红藻活性物质的发现及松节藻醇提物抗糖尿病药理研究

海藻是海洋生物中的一大类群，由于其特殊的生活环境，能够代谢产生大量结构独特多变和活性特殊多样的代谢产物，是化学和生物活性多样性研究的重要对象之一。我国海域辽阔，海藻资源丰富，为寻找结构新颖、生理活性独特的先导化合物，加强对海藻资源的开发利用，本论文对中国沿海的三种海洋红藻进行了化学成分和生物活性研究，同时对山东青岛海域生物量丰富的一种海洋红藻松节藻进行了动物体内抗糖尿病活性研究。 利用正相硅胶柱色谱、Sephadex LH-20柱色谱以及反相HPLC和重结晶等现代分离手段，对山东青岛沿海的红藻扇形叉枝藻（Gymnogongrus flabelliformis）进行了系统的化学成分研究，从中得到单体化合物26个，通过波谱学方法（IR、MS、NMR等）鉴定了他们的结构，分别为(3R,6R,7E)-(+)-3-O-phenylacetyl- 4,7-megastigmadiene-9-one（1），(3R,7E)-(-)-3-O-phenylacetyl-5,7-megastigmadiene -9-one（2），(3S,6R,7E)-(+)-3-hydroxyl-4,7-megastigmadien-9-one（3），(3S,5R,6S,7E)- (-)-3-hydroxy-5,6-epoxy-7-megastigmene-9-one（4），(3S,5S,6R,7E)-(+)-3-hydroxy- 5,6-epoxy-7-megastigmene-9-one（5），Dehydrovomifoliol（6），(3R)-(-)-4-[(2R,4S)-4- acetoxy-2-hydroxy-2,6,6-trimethylcyclohexylidene]-3-buten-2-one（7），2,3,3′-三溴-4,4′,5,5′-四羟基-1′-乙氧甲基双苯基甲烷（8），2,2′,3,3′-四溴-4,4′,5,5′-四羟基双苯基甲烷（9），3-溴-4,5-二羟基苯甲醛（10），2,3-二溴-4,5-二羟基苯甲基甲醚（11），2,3-二溴-4,5-二羟基苯甲醇（12），N, N-二甲基酪胺（13），4-羟基苯甲酸乙酯（14），4-羟基苯甲基乙醚（15），4-羟基苯乙基乙酯（16），4-羟基苯乙酸甲酯（17），4-羟基苯甲醛（18），豆甾-4-烯-3-酮（19），胆甾-4-烯-3-酮（20），胆甾醇（21），尿嘧啶（22），尿嘧啶核苷（23），腺嘌呤核苷（24），丁二酸（25），5-羟基-4-甲基-5-戊基-2,5-二氢呋喃-2-酮（26）。其中化合物1、2为新化合物，化合物3为新天然产物，所有化合物均为首次从该属海藻中分离得到。通过 MTT 法对部分单体化合物进行了肿瘤细胞毒活性筛选, 结果表明，化合物8、9、10、12对筛选的所有细胞株均有较强细胞毒活性，化合物11对人肺癌细胞株（A549）、人肝癌细胞株（Bel 7402）、人结肠癌细胞株（HCT-8）有一定细胞毒活性。通过研究单体化合物对小鼠腹腔巨噬细胞TNF-分泌的影响，对其进行抗炎活性筛选，结果表明，化合物8、9、11、13、17、23、24、25对小鼠腹腔巨噬细胞TNF-分泌表现出明显的抑制作用。 从采自山东荣成镆铘岛的红藻小珊瑚藻（Corallina pilulifera）的乙酸乙酯萃取物中分离得到16个单体化合物，通过波谱学方法鉴定化合物结构14个（另外2个正在鉴定中），分别为2α-乙氧酰基-2β-羟基-A-降胆甾-5-烯-4-酮（27），胆甾-4-烯-3-酮（28），胆甾醇（29），3β-羟基-胆甾-5,24(28)-二烯-7-酮（30），2α-羟基-胆甾-4-烯-3-酮（31），6α-羟基-胆甾-4-烯-3-酮（32），3β-羟基-胆甾-5-烯-7-酮（33），(E)-phytol epoxide（34），Phytenal（35），3,7,11,15- tetramethyl-hexadec-2-en-1-oll（Phytol）（36），Loloilide（37），(3S,5R,6S,7E)-(-)-3-hydroxy-5,6-epoxy-7- megastigmene-9-one（38），Dehydrovomifoliol（39），4-羟基苯甲醛（40）。其中，化合物 31为新天然产物，化合物27为首次从植物中分离得到，所有化合物均为首次从该种海藻中分离得到。通过 MTT 法对分离得到的单体化合物进行了肿瘤细胞毒活性筛选,化合物27和化合物32对筛选的所有肿瘤细胞株均有细胞毒活性，且化合物27对人胃癌细胞株（BGC-823）、人结肠癌细胞株（HCT-8）和人卵巢癌细胞株（A2780）具有中等强度抑制活性。化合物28、化合物31和化合物33对人肝癌细胞株（Bel 7402）、人结肠癌细胞株（HCT-8）和人卵巢癌细胞株（A2780）有一定细胞毒活性。 从采自广西北海涠洲岛的多管藻Polysiphonia sp.的乙酸乙酯萃取物中分离得到6个单体化合物，通过波谱学方法鉴定化合物结构5个（另外1个仍在鉴定），分别为胆甾醇（41），3,7,11,15-tetramethyl-hexadec-2-en-1-ol（Phytol）（42），3-吲哚甲醛（43），4-羟基苯甲醛（44），4-羟基苯甲酸（45）。 对山东青岛沿海的松节藻 (Rhodomela confervoides) 乙醇提取物进行了初步的体内抗糖尿病活性研究，采用链脲佐菌素诱导的2型糖尿病（STZ-DM）大鼠模型对其进行体内降糖实验，结果发现，松节藻乙醇提取物在糖尿病大鼠体内不仅具有显著的降血糖作用，且呈现良好的量–效关系，而且能够纠正糖尿病引发的物质代谢紊乱，增加体重，提高试验动物的成活率，因此具有良好的应用开发前景。

两种海洋生物内生真菌的化学成分及其生物活性研究

海洋微生物次生代谢产物是海洋天然产物研究的重要组成部分。其中海洋真菌因其代谢可以产生大量结构新颖活性独特的化合物，已经成为天然药物的重要来源。近来已经有许多关于海洋生物内生真菌的生理活性次生代谢的报道，为寻找发现新的药物先导化合物提供了可能。本论文对一株毛壳霉属海藻内生真菌Chaetomium sp和一株红树林内生真菌的次生代谢产物的化学成分进行了研究，对其中分离得到的化合物进行了初步的生物活性筛选。 毛壳霉属海藻内生真菌Chaetomium sp.系从青岛近海采集的红藻多管藻Polysiphonia urceolate Grev中分离得到。红树林内生真菌Fs系从采自中国南海红树林植物海桑Sonneratia caseolaris中分离得到。对这两株真菌分别进行了发酵培养，对真菌培养物的菌丝体和发酵液分别用甲醇和乙酸乙脂进行提取。将提取液经减压蒸干后经HPLC检测，两部分基本相同，合并进行化学分离。 对发酵提取物采用常规的硅胶柱层析、制备薄层层析、凝胶Sephadex LH-20柱层析、反相硅胶柱层析，重结晶以及半制备高效液相色谱等分离手段，得到单体化合物。利用各种现代波谱技术(IR、UV、EI-MS、FAB-MS、HR-ESI-MS、1H-NMR、13C-NMR、DEPT、1H-1H COSY、HMQC、HMBC等)，结合甲醇解等化学转化方法鉴定了其中45个化合物的结构。其中，从Chaetomium sp分离鉴定了35个化合物，其中2个为新化合物，分别为化合物EN1 （命名为Chaetopyranin）、EN2 （命名为Chaetofuranin）。从Fs中分离鉴定了10个化合物的结构，其中1个为新化合物Fs1。 对大部分分离到的单体化合物进行了细胞毒活性测试，结果显示新化合物EN1对3株细胞系SMMC-7721, HMEC和A-549具有中等细胞毒活性，IC50分别为28.5, 15.4, 39.1 μg/mL。同时一些已知化合物特别是苯甲醛类衍生物对A-549显示了较强的细胞毒活性，这些在以前文献中未见报道。

Screening marine algae from China for their antitumor activities

Thirty-nine species of marine algae collected from the coast of China were screened for their antitumor activities, and eight species Leathesia difformes, Polysiphonia urcedata, Scytosiphon lomentarius, Gloiopeliis furcata, Punctaria latifolia, Symphyocladia latiuscula, Rhodomela confervoides and Ulva pertusa showed potent cytotoxic activities. Three, Rhodomela confervoides, Scytosiphon lomentarius and Gloiopeliis furcata, were used for further investigation. More than 30 compounds were isolated and purified, and 14 bromophenols, 1 steroid and 1 carotene were identified by advanced spectroscopic methods including IR, MS, and NMR techniques. Amongst the 16 identified compounds, 7 showed vigorously selective activities against KB, Bel7402 and A549 cancer cells, and 6 bromophenols were new compounds.

Life history variation in Dasya ocellata (Dasyaceae, Rhodophyta)

Four of the five members of the Dasyaceae found in the British Isles, Dasya corymbifera J. Agardh, Dasya hutchinsiae Harvey, Dasya punicea Meneghini ex Zanardini and Heterosiphonia plumosa (Ellis) Batters, appear to have Polysiphonia-type life histories on the basis of evidence from field collections of tetrasporophytes and gametophytes. In collections from the British Isles of the fifth species, Dasya ocellata (Grateloup) Harvey, only tetrasporophytes have ever been observed, but there are two reports of gametophytes in this species from further south in Europe. Dasya ocellata tetraspores were isolated into culture from populations in Strangford Lough, Northern Ireland, and Agadir, Morocco, where one female thallus was collected amongst tetrasporophytes. Dasya ocellata from Ireland underwent a direct tetraspore-to-tetrasporophyte life history, which was followed through two complete cycles. Karyological studies showed that meiosis does not occur during tetrasporangial development: tetrasporangia are mitotic, with c. 64 small chromosomes. Comparison with chromosome numbers in meiotic tetrasporangia of D. hutchinsiae (n = c. 32) showed that this is the diploid chromosome complement. Tetraspores from the Moroccan isolate, by contrast, gave rise to gametophytes (although only the males became fertile) and tetrasporophyte recycling did not occur. Thalli sampled from a population in southern Portugal consisted only of tetrasporophytes. Dasya ocellata, like many members of the Ceramiales, shows intraspecific life history variability; a sexual life history apparently occurs only in southern populations.

Flora polínica da Reserva do Parque Estadual das Fontes do Ipiranga (São Paulo, Brasil): família: 135-Asclepiadaceae

No presente trabalho, foram estudados os polinários de nove gêneros e 16 espécies de Asclepiadaceae ocorrentes na Reserva do Parque Estadual das Fontes do Ipiranga: Asclepias curassavica L., Blepharodon nitidum (Vell.) J F. Macbr., Ditassa burchellii var. vestita (Malme) Fontella, Ditassa hispida (Vell.) Fontella, Ditassa tomentosa (Decne.) Fontella, Gomphocarpus physocarpus E. Mey., Gonianthela axillaris (Vell.) Fontella & E.A. Schwarz, Matelea glaziovii (E. Fourn.) Morillo, Orthosia urceolata E. Fourn., Oxypetalum appendiculatum Mart., Oxypetalum capitatum subsp. capitatum Mart., Oxypetalum insigne (Decne.) Malme, Oxypetalumpachyglossum Decne., Oxypetalumpedicellatum Decne., Oxypetalum wightianum Hook. & Arn., Tassadia subulata var. subulata (Vell.) Fontella & E.A. Schwarz. São apresentadas descrições e ilustrações para todas as espécies estudadas, bem como chaves para identificação das espécies dos gêneros Ditassa e Oxypetalum tomando como base a morfologia dos polinários.

Factors structuring Fucus communities at open and complex coastlines in the Baltic Sea

This thesis deals with physical factors and biological interactions affecting the distribution of two fucoid species, Fucus vesiculosus and F. serratus, in the Baltic Sea. Studies have been carried out in two quite different environments: an archipelago, and an open rocky coast. The archipelago has an extremely long coastline with a heterogeneous submerged landscape of different substrate types, slopes, water qualities, and degrees of wave exposure. The factors influencing F. vesiculosus distribution, morphology and epiphyte composition were studied in the Stockholm archipelago using field surveys and spatial modelling in Geographic information systems (GIS). A GIS-method to estimate wave exposure was developed and validated by comparing the result to an index based on vertical zonation of lichens. Wave exposure was considered an important factor for predicting the distribution of F. vesiculosus by its ability to clean hard surfaces from silt, and a predictive model was constructed based on the information of wave exposure and slope of the shore. It is suggested that the lower distribution boundary of attached F. vesiculosus is set by sediment in sheltered parts of the archipelago, and by light availability in highly wave exposed parts. The morphology of F. vesiculosus was studied over a wave exposure gradient, and several characters responded in accordance with earlier studies. However, when separating effects of wave exposure from effects of other confounding water property parameters, only thallus width was significantly different. Several water property parameters were shown to be correlated with wave exposure in the Stockholm archipelago, and the mechanism responsible for the effects on F. vesiculosus morphology is discussed. The composition of epiphytes on F. vesiculosus varied over a wave exposure gradient with a positive correlation to Elachista fucicola, and a negative to Chorda filum. At an open coast the physical environment is much less heterogeneous compared to an archipelago. The distributions of F. vesiculosus, F. serratus, turf-forming algae, and the seafloor substrate, were surveyed along the open coasts of Öland and Gotland. Turf-forming algae dominated all hard substrates in the area, and Polysiphonia fucoides was most abundant. At the Gotland coast F. vesiculosus was less abundant than at the Öland coast, and F. serratus occurred only in the southern-most part. Fucus serratus was increasingly more common towards south which was interpreted as an effect mainly of the Baltic salinity gradient, or the variation of salinity that has occurred in the past. The effects of turf-forming algae and sediment on F. serratus recruitment at 7 m depth off the Öland east coast were studied in the field, and by laboratory experiments. Almost no recruits were found in the algal turf outside the F. serratus patches. More fine sediment was found in the turf than in the F. serratus patches, suggesting that the turf accumulates sediment by decreasing resuspension. Both filamentous algae and sediment decreased the attachment ability of F. serratus zygotes and survival of recruits, and sediment had the strongest effect. It is therefore suggested that F. serratus has difficulties recruiting outside its patches, and that these difficulties are enforced by the eutrophication of the Baltic Sea, which has favoured growth of filamentous algae and increased sedimentation. An overall conclusion is that Fucus distribution is affected by large-scale-factors, such as the eutrophication and salinity changes of the Baltic Sea, as well as by small-scale variation in wave exposure, substrate and slope, and by surface competition with neighbouring species.

Epibiotic assemblages on seaweeds in the German Wadden Sea (North Sea)

After detachment from benthic habitats, the epibiont assemblages on floating seaweeds undergo substantial changes, but little is known regarding whether succession varies among different seaweed species. Given that floating algae may represent a limiting habitat in many regions, rafting organisms may be unselective and colonize any available seaweed patch at the sea surface. This process may homogenize rafting assemblages on different seaweed species, which our study examined by comparing the assemblages on benthic and floating individuals of the fucoid seaweeds Fucus vesiculosus and Sargassum muticum in the northern Wadden Sea (North Sea). Species richness was about twice as high on S. muticum as on F. vesiculosus, both on benthic and floating individuals. In both seaweed species benthic samples were more diverse than floating samples. However, the species composition differed significantly only between benthic thalli, but not between floating thalli of the two seaweed species. Separate analyses of sessile and mobile epibionts showed that the homogenization of rafting assemblages was mainly caused by mobile species. Among these, grazing isopods from the genus Idotea reached extraordinarily high densities on the floating samples from the northern Wadden Sea, suggesting that the availability of seaweed rafts was indeed limiting. Enhanced break-up of algal rafts associated with intense feeding by abundant herbivores might force rafters to recolonize benthic habitats. These colonization processes may enhance successful dispersal of rafting organisms and thereby contribute to population connectivity between sink populations in the Wadden Sea and source populations from up-current regions.

Quaternary radiolarian counts from Site 181-1123

In the present work Quaternary radiolarian assemblages from the Southwest Pacific were investigated due to their importance for correlation and identification of climatic changes. The studied Ocean Drilling Program (ODP) Site 1123 (Leg 181) is situated on the northern flanks of the Chatham Rise, 1100 kilometres offshore eastern New Zealand and in a water depth o f 3290 metres. It is situated just north of the Subtropical Convergence (STC) in temperate climatic conditions, influenced by the cold deep Deep Western Boundary Current (DWBC) and by the subtropical East Cape Current (ECC) in shallow water depths. A continuous record of 79 sediment samples from this site with a temporal resolution of ~15,000 years provided a medium-resolution record of radiolarian assemblages through the Quaternary. This allowed investigations on how radiolarian assemblages are influenced by climatic variations at obliquity and eccentricity bandwidth, with periodic variations of 40,000, 100,000 and 400,000 years, respectively. Emphasis was given to changes in radiolarian assemblages through the Mid-Pleistocene climate transition (MPT) that marks a fundamental reorganisation in Earth's climate system by change from 40,000 to 100,000 year cycles. Glacial and interglacial variations in oceanography were investigated. Especially the influence of the DWBC was examined due to its input of deep and cold waters to the Pacific Ocean, which plays an important role in Earth's climate system. 167 radiolarian counting groups were examined concerning variations in radiolarian abundance, preservation, diversity, the relative abundance of orders, families, and selected species in order to detect influences of past climatic variations in the Southwest Pacific. No significant changes in radiolarian assemblages were found in coincidence with the onset of the MPT. Investigations led to the recognition of four characteristic phases within the last 1.2 million years. Within one of these phases (Phase Ill), about 160,000 years after the onset of the MPT, fundamental changes in radiolarian assemblages occurred. Investigations yielded highest diversity and highest numbers of nassellarians in abundant samples, whereas sparse samples were mostly poorly preserved and were dominated by spumellarians. Abundance of certain radiolarian families in interglacials or glacials indicated their usefulness as indicators for climatic conditions at Site 1123. Trends o f selected taxa within these families supported the significance of warm- or cool-water preference of these families. Use of 67 radiolarian species as climate indicators showed abundance of warm-water assemblages within interglacials, whereas abundance of cool-water species was increased within glacials. Depth distributional patterns of 52 radiolarian species indicated a strong influence of shallow waters, possibly the EEC, within interglacials and increased influence of deep and intermediate waters, possibly of southern-sourced character and the DWBC in glacial stages.

Radiolarians from ODP Leg 116 holes

Cores recovered from three sites of Leg 116 were studied for radiolarians. Generally, radiolarians were absent from most samples prepared for examination. Moderate to well-preserved radiolarian assemblages are found only in the uppermost one or two cores that were the focus of this study. All of the radiolarian assemblages in the upper cores belong to the Buccinosphaera invaginata Zone of latest Quaternary age. However, there is one stratum where a few Miocene radiolarians are reworked into the modern assemblages. Local seamounts are suggested sources for the reworked radiolarians.