利用三种分子标记研究缘毛类纤毛虫的系统发育地位

为了探讨缘毛类纤毛虫的系统发育地位 ,利用RAPD方法得到了 9种缘毛类纤毛虫、 1种四膜虫和1种喇叭虫的 3个随机引物的电泳带谱 ;测定了 7种缘毛类纤毛虫rRNA基因中的间隔区 1(ITS1)和小亚基核糖体核糖核酸 (SSrRNA)基因序列 ,并构建了相应的系统树。在比较和分析RAPD、ITS1和SSrRNA基因序列在缘毛类纤毛虫系统发育研究中的适用范围的基础上 ,以SSrRNA基因序列为分子标记研究了缘毛类纤毛虫系统发育地位 ,结果表明 :①缘毛亚纲是单系的 ,作为寡膜纲中一个亚纲的分类地位是合理的 ;②缘毛类纤毛虫可能是寡膜纲中较高等的一个类群。     

游仆目纤毛虫(纤毛门,旋唇纲)的分子系统学研究

在最新GenBank分子信息的汇集、分析基础上,利用小亚基单位核糖体RNA基因序列分别构建了最大简约树、距离树、最大似然树及贝叶斯树,对长期存在争议的游仆目纤毛虫的系统关系进行了探讨和修订.工作显示:1)游仆目不是1个单源发生系;2)盘头虫类从游仆目中较早分化出来,呈现与排毛类、寡毛类平行的进化关系;3)腹棘虫科内的腹棘虫和拟游仆虫体现了密切的亲缘关系;4)游仆虫科、舍太虫科、檐纤虫科在所有分子树中均表现了十分稳定的拓扑结构;5)游仆虫属内可以细分为7个亚类群;6)尾刺虫科在游仆目内分化较早,支持前人有关其在系统演化时序中处于原始阶段的论断.     

泽蛙原蛙片虫的形态学研究

对采自湖北省洪湖地区的泽蛙(Fejervarya limnocharis)肠道内寄生的泽蛙原蛙片虫进行了形态学再描述, 包括活体形态、固定染色标本形态及扫描电镜下超微结构。样本形态特征参数值与Nie(倪达书)首次发现并命名时所记述标本参数值颇为相符(包括体长、体宽, 核长、核宽)。此外, 本文对2个重要的分类特征进行了补充和修正: (1)缝线位于虫体顶端, 贯穿背腹侧; 所有体动基列均从缝线两侧发出。(2)胞核分裂时在两新核间产生的连接细丝, 在虫体整个生活周期一直存在。并针对这2点特征与其他已知的原蛙片虫属种类进行了比较和讨论: 认为缝线结构是较为稳定的分类特征之一, 体动基列均由此处生发并与虫体纵轴平行延伸; 推测泽蛙原蛙片虫子代虫体中的2个胞核均来自于母体中同一个核, 而其2核间的连接细丝可能是胞核分裂进化历程中比较原始的残迹。     

土霉素对扇形游仆虫(原生生物)种群生长及遗传多样性的影响

抗生素作为一类环境污染物在海岸带生境中广泛存在,它们对海洋原生生物遗传多样性的影响还不清楚.本工作以一种海洋纤毛虫原生生物(扇形游仆虫(Euplotes vannus))为例,研究了一种常见四环类抗生素(土霉素)对其生长的影响,并通过单细胞(个体)分析,探索了抗生素对其大核基因组中核糖体小亚基RNA基因(SSU r DNA)序列多样性的影响.结果表明,与对照组相比,3个土霉素处理组(1,10和20μg/m L)中纤毛虫的自然增长率随土霉素浓度升高显著降低,细胞大小则呈增大趋势.SSU r DNA序列单倍型多样性、核苷酸多样性和GC含量随抗生素浓度升高呈现降低趋势.序列单核苷酸多态性位点的碱基转换频率远高于颠换,但随土霉素浓度升高,颠换频率呈增加趋势.在20μg/m L处理组中,SSU r DNA序列中胸腺嘧啶(T)突变为胞嘧啶(C)的频率显著升高,而腺嘌呤(A)突变为鸟嘌呤(G)的频率显著降低.这表明,海岸带抗生素污染对纤毛虫表型与生理产生显著影响的同时也导致基因组内r DNA遗传结构的变化,显示纤毛虫原生生物可通过基因组高度的可塑性来快速适应抗生素胁迫.     

利用核糖体DNA限制性酶切谱(ARDRA)分子技术和小亚基单位核糖体RNA(SSrRNA)基因序列分析对五种双眉虫(原生动物,纤毛门,游仆目)种内及种间关系的探讨

从12种限制性内切酶中筛选出6种可获得种间多样性的内切酶:Alu Ⅰ,Taq Ⅰ,Hae Ⅲ,Hinf Ⅰ,Msp Ⅰ,Xba Ⅰ,对广义双眉虫Diophrys-complex5个种(伪寡毛双眉虫Diophrys apoligothrix、悬游双眉虫D.appendiculata、盾圆双眉虫D.scutum、秀丽拟双眉虫Paradiophrys irmgard、针毛类双眉虫Diophryopsis hystrix)共7个种群的核糖体基因(18S小亚基、部分23S大亚基及其内转录间隔区域)进行多位点酶切。结果显示,种间差异明显大于种内差异。利用RAPDistance1.04软件构建的邻接树表明,盾圆双眉虫的3个种群表现出高度的同源性;3个近缘属(双眉虫属、拟双眉虫属、类双眉虫属)可以被明确区分,并支持类双眉虫属与拟双眉虫属的独立性。从GenBank/EMBL数据库中获得广义双眉虫及相近种的小亚基单位核糖体RNA(SSrRNA)基因序列,利用邻接法(NJ),贝叶斯法(Bayesian)和最大简约法(MP)构建的系统发生树具有基本一致的拓扑结构,结果显示:狭义双眉虫属Diophrys为单源发生系,并与类双眉虫属Diophryopsis组成姐妹群;尽管拟双眉虫属Paradiophrys具有广义双眉虫典型的形态学特征,但与尾刺虫属Uronychia有着较近的亲缘关系。本工作同时表明,核糖体DNA限制性酶切(ARDRA)技术可靠地区分纤毛虫的形态相似种,并在双眉虫属间水平的系统关系推定中存在一定的适用性。     

尖前口虫的口器发生研究(纤毛门, 膜口目)

研究了咽膜类纤毛虫尖前口虫无性生殖期的核器及口器的演化.其发生特征为;1)新的口原基形成于原前庭动基列与口侧膜间,表观为原口侧膜分裂而致;2)随着形态发生的进行,由口原基依次演化出后仔虫的三片咽膜、口侧膜和三条前庭动基列;3)原口器完全被前仔虫所继承;4)体纤毛器在整个形态发生过程中一直保持双动基列结构.       

盾圆双眉虫与伪寡毛双眉虫的形态学重描述(原生动物,纤毛门)

研究对采自青岛沿海的两种海洋纤毛虫-盾圆双眉虫与伪寡毛双眉虫做了形态学重描述。盾圆双眉虫与前人所报道的种群具有十分相似的纤毛图式,但在额-腹棘毛分布、大核片段、小膜及背触毛数目等方面表现出细微的变异性。此外,该青岛种群个体较小。统计学比较还表明,迄今缺乏研究的一海洋种,泥生双眉虫极可能为盾圆双眉虫(Diophrys scutum)的同物异名。伪寡毛双眉虫(Diophrys apoligothrix)为一新近报道的罕见种,研究基于新采集种群对其进行了补充性观察和描述。       

五种缘毛类纤毛虫遗传关系的ISSR研究

应用ISSR分子标记揭示了5种缘毛类纤毛虫(Carchesium polypinum,Epistylis chrysemydis,E.plicatilis,E.urceolata和Vorticella campanula)的遗传关系.从34个引物中筛选到13个多态性高的引物进行研究.得到的遗传距离(0.666 7～1.000 0)显示ISSR技术具有较高的分辨率.依据构建的UPGMA聚类树,V.campanula首先和其他种类分开;C.polypinum和E.chrysemydis聚在了一起;在3种Epistylis纤毛虫中,E.plicatilis和E.urceolata聚在了一起.对比由核糖体小亚基RNA基因序列构建的系统发育树,发现:1)ISSR引物在缘毛类纤毛虫基因组中可以得到多态扩增;2)C.polypinum与Epistylis的关系近于V.campanula,在缘毛类纤毛虫分类中,单生或群居是重要的系统发育特征;3)E.plicatilis和E.urceolata的关系近于E.chrysemydis.在Epistylis属纤毛虫中,柄的中空与否是一个有用的系统发育和分类特征.本研究表明ISSR方法在纤毛虫相近和相似种遗传关系研究中是一种新的有用方法.     

小腔游仆虫形态学、个体发育与分子系统学研究

游仆类是纤毛虫中进化最为复杂和高等的一大类群,为了进一步探索和完善游仆类的多样性,本研究利用活体观察、蛋白银和银浸法染色技术对采自青岛小西湖的小腔游仆虫(Euplotes aediculatus)的形态学及细胞发生学进行了详尽的研究,并在完整的形态学及发生学研究基础上,测定了小腔游仆虫的核糖体小亚基基因(SSU r DNA)序列,通过序列比较和分子系统树构建等方法,对小腔游仆虫的系统地位进行了分析。结果表明:本种鉴别特征为9根额腹棘毛,5根横棘毛,2根缘棘毛,2根尾棘毛,8列背触毛,double-eurystomus型银线系。发生学特征包括:(1)后仔虫口原基在表皮下独立发生,前仔虫完全继承老口围带;(2)额–腹–横棘毛原基从左向右按照3:3:3:2:2的模式形成额腹棘毛和横棘毛;(3)前后仔虫最左侧额腹棘毛分别由独立产生的原基形成;(4)缘棘毛原基独立发生;(5)初级背触毛原基来自虫体中部老结构的反分化;(6)前后仔虫尾棘毛分别来自最右侧2列背触毛原基和老背触毛列末端;这些特征显示出游仆虫属个体发生模式的高度保守性。分子系统分析与形态学数据一致,即游仆虫属为单元发生,且小腔游仆虫与艾美游仆虫(Euplotes amieti)、阔口游仆虫(E.eurystomus)和伍氏游仆虫(E.woodruffi)聚在一起。     

广东大亚湾六种海洋刺钩类纤毛虫的形态学研究

利用活体观察及银染法首次对采自广东大亚湾沿岸的盐菲阿虫Phialina salinarum Kahl,1928,海洋长吻虫Lacrymaria marina Kahl,1933,惊扰伪颈毛虫Pseudotrachelocerca trepida(Kahl,1928)Song,1990,贪食纤口虫Chaenea vorax Quennerstedt,1867,柱核柱毛虫Cyclotrichium cyclokaryon Meunier,1907和加冈栉毛虫Didinium gargantua Meunier,1910六种刺钩类纤毛虫进行了形态学研究,补充了活体形态学以及纤毛图式等分类学新信息。首次观察到了海洋长吻虫体纤毛基部的皮层颗粒和柱核柱毛虫大亚湾种群较青岛种群具有更多的体动基列等若干新信息。研究表明,刺钩类纤毛虫在大亚湾近岸水域具有较高的物种多样性。       

GRACILARIOPSIS MCLACHLANII SP. NOV. AND GRACILARIOPSIS PERSICA SP. NOV. OF THE GRACILARIACEAE (GRACILARIALES,RHODOPHYCEAE) FROM THE INDIAN OCEAN1

Two new species of Gracilariopsis from the Indian Ocean are proposed—Gracilariopsis (Gp.) mclachlanii Buriyo, Bellorin et M. C. Oliveira sp. nov. from Tanzania and Gracilariopsis persica Bellorin, Sohrabipour et E. C. Oliveira sp. nov. from Iran—based on morphology and DNA sequence data (rbcL gene and SSU rDNA). Both species fit the typical features of Gracilariopsis: axes cylindrical throughout, freely and loosely ramified up to four orders, with an abrupt transition in cell size from medulla to cortex, cystocarps lacking tubular nutritive cells and superficial spermatangia. Nucleotide sequence comparisons of rbcL and SSU rDNA placed both species into the Gracilariopsis clade as distinct species from all the accepted species for this genus, forming a deeply divergent lineage together with some species from the Pacific. The new species are very difficult to distinguish on morphological grounds from other species of Gracilariopsis, stressing the importance of homologous molecular marker comparisons for the species recognition in this character‐poor genus.     

多节核棘尾虫的发现及其与邻近属种的形态比较

黑龙江省帽儿山镇沼泽地生存着一种在体形和皮层形态上酷似贻贝棘尾虫的腹毛类纤毛虫,其大核呈多个结节相连的索状,这与传统的棘毛虫属的特征有别,但S期大核复制带两个仍在两端出现,这一点全似棘尾虫属,又鉴于腹毛类的其它属中也有大核结数目不同的种（Borror,1983),故单纯大核结节数目不同不应被视为属间判别,因此本文将此虫定为棘尾虫属一新种,名之为多节核棘尾虫,据此,棘尾虫属的传统鉴别特征在核形上也予以修正。     

AGLAOTHAMNION HALLIAE COMB. NOV. AND A. COLLINSII SP. NOV. (CERAMIALES, RHODOPHYTA): RESOLUTION OF NOMENCLATURAL AND TAXONOMIC CONFUSION

Specimens from the exsiccatae Phycotheca BorealiAmericana number 698 are shown to represent two distinct species. One of them, formerly known as Callithamnion halliae, is transferred to Aglaothamnion on the basis of its uninucleate cells. The specimens exhibit distichous branching and are ecorticate. A lectotype is proposed for one of the P.B.-A. #698 specimens displaying these characters . Aglaothamnion halliae ( Collins) comb. nov. is an earlier name for Aglaothamnion westbrookiae Rueness et L'Hardy-Halos. The second species, together with specimens of P.B.-A. #1896, has radial branching and corticated axes and is described herein as a new species , Aglaothamnion collinsii. The binomial Aglaothamnion boergesenii ( Aponte et Ballantine ) L'Hardy-Halos et Rueness in Aponte, Ballantine, et J. Norris is validated. A key to the tropical and warm-temperate western Atlantic species of Aglaothamnion is presented .     

ULTRASTRUCTURE AND 18S RRNA GENE SEQUENCE FOR PELAGOMONAS CALCEOLATA GEN. ET SP. NOV. AND THE DESCRIPTION OF A NEW ALGAL CLASS,THE PELAGOPHYCEAE CLASSIS NOV.1

Pelagomonas calceolata gen. et sp. nov., an ultra-planktonic marine alga, is described using electron microscopy and the cytoplasmic small subunit (18S) ribosomal RNA (rRNA) gene sequence. Cells are uniflagellate, about 1.5 × 3 μm in size. The flagellium has two rows of bipartite hairs, the paraxonemal rod has a dentate appearance, and a two-gyred transitional helix is present between two transitional plates. Microtubular roots, striated roots, and a second basal body are absent. A thin organic theca surrounds most of the cell. There is a single chloroplast with a girdle lamella and a single, dense mitochondrion with tubular cristae. A single Golgi body with swelled cisternae lies beneath the flagellum, and each cell has an ejectile organelle that putatwely releases a cylindrical structure. A vacuole, or cluster of vacuoles, contains the putative carbohydrate storage product. The 18S rRNA gene was sequenced completely in both directions, excluding three primer regions. When compared to the same gene sequence from other organisms, Pelagomonas calceolata gen. et sp. nov. occupies an unresolved position among other chromophyte algae and is distinct from members of any of these classes. Based on morphological, ultrastructural, and molecular data, we describe this alga as a new species, and we place this highly unusual new species in a new genus, family, order, and class.     

STRUCTURE AND REPRODUCTION OF AMANSIA AND MELANAMANSIA GEN. NOV. (RHODOPHYTA,RHODOMELACEAE) ON THE SOUTHEASTERN AFRICAN COAST1

Four species of Amansia Lamouroux were initially found in Natal. More Complete studies on these species revealed a new genus, Melanamansia, Which is described on the basis of presence of two dorsal pseudopericentral cells in two new species from Natal (M. seagriefii sp. nov. & M. fimbrifolia sp. nov.) in addition to other structural characters and features of pigmentation and reproduction. Pseudopericentral cells are not present in the type species of Amansia, A. multifida Lamouroux. The other two species of Amansia occurring in Natal, A. glomerata C. Agardh & A. loriformis sp. nov., have characters similar to the type species. Comparison of species from other regions of the world has shown that eight additional species, previously assigned to Amansia, belong to the new genus.     

洞庭湖外睾吸虫新种及其生活史

本文报告洞庭湖区鲶鱼肠道寄生的洞庭湖外睾吸虫Exorchis dongtinghuensis sp.nov(新种)及其全程生活史,其第一中间宿主为湖北钉螺Oncomelania hupensis;第二中间宿主为鲤鱼、鲫鱼和金鱼;终宿主为鲶鱼Parasilurus asotus。作者对各期宿主作了人工感染试验和现场自然感染调查。对其发育过程作了观察比较。     

斑痣盘菌科一新种——松生小鞋孢盘菌

本文报道在云南省昆明市华山松(Pinus armandi Franch.)上发现的斑痣盘菌科一新种,即松生小鞋孢盘菌(Soleella pinicola Y.R.Lin et W.Ren)。对该种作了拉丁文、汉文描述和图解。主模式标本保藏于安徽农学院林学系森林保护教研室。     

MORPHOLOGY AND MOLECULAR ANALYSES OF THREE TOXIC SPECIES OF GAMBIERDISCUS (DINOPHYCEAE): G. PACIFICUS, SP. NOV., G. AUSTRALES, SP. NOV., AND G. POLYNESIENSIS, SP. NOV.

Three new dinoflagellate species, Gambierdiscus polynesiensis, sp. nov., Gambierdiscus australes, sp. nov., and Gambierdiscus pacificus, sp. nov., are described from scanning electron micrographs. The morphology of the three new Gambierdiscus species is compared with the type species Gambierdiscus toxicus Adachi et Fukuyo 1979, and two other species: Gambierdiscus belizeanus Faust 1995 and Gambierdiscus yasumotoi Holmes 1998. The plate formula is: Po, 3′, 7", 6C, 8S, 5‴, 1p, 2". Culture extracts of these three new species displayed both ciguatoxin- and maitotoxin-like toxicities. The following morphological characteristics differentiated each species. 1) Cells of G. polynesiensis are 68–85 μm long and 64–75 μm wide, and the cell’s surface is smooth. They are identified by a large triangular apical pore plate (Po), a narrow fish-hook opening surrounded by 38 round pores, and a large, broad posterior intercalary plate (1p) wedged between narrow postcingular plates 2‴ and 4‴. Plate 1p occupies 60% of the width of the hypotheca. 2) Cells of G. australes also have a smooth surface and are 76–93 μm long and 65–85 μm wide in dorsoventral depth. They are identified by the broad ellipsoid apical pore plate (Po) surrounded by 31 round pores and a long and narrow 1p plate wedged between postcingular plates 2‴ and 4‴. Plate 1p occupies 30% of the width of the hypotheca. 3) Cells of G. pacificus are 67–77 μm long and 60–76 μm wide in dorsoventral depth, and its surface is smooth. They are identified by the four-sided apical pore plate (Po) surrounded by 30 round pores. A short narrow 1p plate is wedged between the wide postcingular plates 2‴ and 4‴. Plate 1p occupies 20% of the width of the hypotheca. These three newly described species were also characterized by isozyme electrophoresis and DNA sequencing of the D8–D10 region of their large subunit (LSU) rRNA genes. The consistency between species designations based on SEM microscopy and classification inferred from biochemical and genetic heterogeneities was examined among seven isolates of Gambierdiscus. Their classification into four morphospecies was not consistent with groupings inferred from isozyme patterns. Three molecular types could be distinguished based on the comparison of their LSU rDNA sequences. Although G. toxicus TUR was found to be more closely related to G. pacificus, sp. nov. than to other G. toxicus strains, the molecular classification was able to discriminate G. polynesiensis, sp. nov. and G. australes, sp. nov. from G. toxicus. These results suggest the usefulness of the D8–D10 portion of the Gambierdiscus LSU rDNA as a valuable taxonomic marker.     

MOLECULAR PHYLOGENY AND TAXONOMY OF THE AEGAGROPILA CLADE (CLADOPHORALES,ULVOPHYCEAE), INCLUDING THE DESCRIPTION OF AEGAGROPILOPSIS GEN. NOV. AND PSEUDOCLADOPHORA GEN. NOV.1

The Aegagropila clade represents a unique group of cladophoralean green algae occurring mainly in brackish and freshwater environments. The clade is sister to the species‐rich and primarily marine Cladophora and Siphonocladus lineages. Phylogenetic analyses of partial LSU and SSU nrDNA sequences reveal four main lineages within the Aegagropila clade, and allow a taxonomic reassessment. One lineage consists of two marine ‘Cladophora’ species, for which the new genus Pseudocladophora and the new family Pseudocladophoraceae are proposed. For the other lineages, the family name Pithophoraceae is reinstated. Within the Pithophoraceae, the earliest diverging lineage includes Wittrockiella and Cladophorella calcicola, occurring mainly in brackish and subaerial habitats. The two other lineages are restricted to freshwater. One of them shows a strong tendency for epizoism, and consists of Basicladia species and Arnoldiella conchophila. The other lineage includes Aegagropila, Pithophora and a small number of tropical ‘Cladophora’ species. The latter are transferred to the new genus Aegagropilopsis. Previously, polypyramidal pyrenoids had been suggested to be apomorphous for this clade, but we report the finding of both polypyramidal and bilenticular pyrenoids in members of the Pithophoraceae, and thus show that this character has no diagnostic value.