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。       

十种旋唇纲纤毛虫的形态学描述

借助活体观察和蛋白银染色方法对采集于中国山东、广东沿海的潮间带、红树林及淡水生境的10种旋唇纲纤毛虫(拉氏伪小双虫Pseudoamphisiella lacazei、四核伪小双虫Pseudoamphisiella quadrinucleata、中华偏全列虫Apoholosticha sinica、维尔伯特泡毛虫Tunicothrix wilberti、海洋后尾柱虫Metaurostylopsis marina、偏澳大利亚速体虫Notohymena apoaustralis、偏寡毛双眉虫Diophrys apoligothrix、寡毛双眉虫Diophrys oligothrix、盾圆双眉虫Diophrys scutum、张氏拟双眉虫Paradiophrys zhangi)进行了活体形态与纤毛图式水平的分类学研究。其中偏澳大利亚速体虫、偏寡毛双眉虫、张氏拟双眉虫为广东地区新记录种。基于新种群, 对张氏拟双眉虫给出了新定义。研究表明, 中华偏全列虫地理分布较广, 可适应从淡水到不同盐度的咸水生境。首次发现, 寡毛双眉虫可在盐度较低的咸水中生存, 显示出其较强的环境适应能力。     

利用RAPD技术对三种伪角毛虫及五种全列虫(原生动物,纤毛虫)的属间及属内关系探讨

利用11种随机引物对腹毛类纤毛虫中两个相近属内8个海洋种(3种伪角毛虫和5种全列虫)进行了种间及种内关系的RAPD分析,所得清晰条带显示种间均具有明显的差异.计算和比较了种群间、同属种间及不同属间的相似性指数.利用RAPDistance软件构建的邻接(NJ)树表明:伪角毛虫属与全列虫属之间能很好地分开,而不同种群间表现了很好的同源性.本工作表明RAPD技术可以很可靠的用来区分形态相似种,同时也表明RAPD分析在纤毛虫属间水平的系统关系推定中存在一定的适用性.     

下毛目分子系统进化研究和鬃棘尾虫再描述(纤毛动物门:下毛目:尖毛虫科)

作者自武汉东湖采集的样本中分离出鬃棘尾虫Stylonychia pustulata,利用活体观察与蛋白银染色技术进行了形态学研究,并测定了核糖体小亚基RNA基因全序列;以邻接法（Neighbor-Joining,NJ)和最大简约性法（Maximum Parsimony Method,MP)对该物种与GenBank中所有其它下毛目物种进行了分子系统发育分析。结果显示：所研究下毛目种类皆聚为一支,支持下毛目为单系观点;尖毛虫属和棘尾虫属物种聚类相互交错,因此结合形态学上分类标志的分析,对这两个属存在合理性提出异议,根据本文对同种不同种群的小腔游仆虫Euplotes aedicalatus、尖毛虫Oxytricha granulifera以及鬃棘尾虫Stylonychia pustulata分子进化分析。说明在研究系统问题时同时给出模式种的形态学及分子生物学两方面资料的必要性。     

懒猴属的核糖体DNA变异及其种间分化关系

用１５种限制性内切酶和人２８Ｓ、１８ＳｒＤＮＡ探针构建了懒猴属各物种核糖体ＤＮＡ重复单位的限制性内切酶图谱。在进化速率较高的非转录间隔区,在大、中、小懒猴中分别定位了２３、２４、２４个酶切位点。大懒猴与中懒猴有１２个位点不同,与小懒猴有１４个位点不同,而中、小懒猴间则只有一个位点的差异。经过计算,大懒猴与中懒猴的遗传距离值为１２．６５％,与小懒猴的差异为１４．２４％,中、小懒猴间的差异则仅为０．７     

中国(虫穴)螨科二新种记述(蜱螨亚纲:中气门亚目)

记述(虫穴)螨科二新种:花副(虫穴)螨Parazercon floralis sp.nov.和拟重卡(虫穴)螨Caurozercon duplexoideus sp.nov.,副(虫穴)螨属和卡(虫穴)螨属为中国首次发现.     

鲈鱼寄生逆转指环虫的分类地位

采用单个虫体PCR扩增、序列测定与分析的方法,对寄生于鲈鱼及鲤科鱼类的4种指环虫的核糖体28SrRNA基因5'端部分序列和内转录间隔区1(ITS1)进行了研究,并采用PAUP软件中的最大简约法(most parsimo-ny,MP)和邻接法(neighborjoining,NJ)构建了28S rDNA分子系统树.结果显示,逆转指环虫明显地嵌合于其它3种指环虫之间,不宜将其从指环虫属中移出另立似小突虫属,从而进一步明确了逆转指环虫的分类地位.     

蓖麻蚕(Attacus ricini)核糖体RNA基因片段在大肠杆菌中的无性繁殖

利用质粒pBR322作运载体,获得了蓖麻蚕(Attacus ricini)核糖体rRNA基因(rDNA)的部分片段在E.coli中的无性繁殖株。酶切图谱分析及Southern法分子杂交鉴定证明,重组质粒pARI含有1.95MdrDNA EcoRI-BamHⅠ双酶切片段;pARⅡ含有2.6Md的rDNABamHⅠ片段。并测定了BamHⅠ片段与pBR322连接方向。     

利用核糖体小亚基RNA基因序列对环须虫属,突口虫属及旋口虫属(原生动物,纤毛门,异毛纲)系统地位的探讨

在系统发育研究中,异毛纲纤毛虫普遍被认为是一个多源发生系.为进一步探讨纲中各科/属间的亲缘关系,新测定了隶属于异毛纲的2属4种的核糖体小亚基基因序列,并结合GenBank中迄今所知该类群的基因序列,利用最大似然法、贝叶斯法、最大简约法和邻接法构建了系统发生树.结果显示:1)所涉及的9个科中,赭纤虫科Blepharismidae、喇叭纤虫科Stentoridae、瓶囊虫科Folliculinidae和缘棘虫科Peritromidae为单源发生,爽口虫科Climacostomidae、旋口虫科spirostomidae和突口虫科Condylostomatidae为非单源发生;2)环须虫属Peritromus与爽口虫属Climacostomum两个属为姐妹类群;3)旋口虫属跏Spirostomum先于其它异毛类分化出去,与格鲁博虫属Gruberia亲缘关系较远;4)突喇叭虫属Condylostentor与突口虫属Condylostoma聚在一起并与夏通属Chattonidium具有更近的亲缘关系,而与喇叭虫属Stentor关系较远.     

基于18S rRNA基因部分序列探讨四种蛙类多盘科吸虫分类地位

分析了寄生于蛙类膀胱的4种多盘虫科Polystomatidae吸虫:石林双睾吸虫Diplorchis shilinensis、锯腿树蛙多盘吸虫Polystoma carvirostris和分别寄生于昭觉林蛙Rana chaochiaoensis、华西雨蛙Hyla a.annectans的2未定种的分类地位,并利用18S rRNA基因部分序列进行了系统发育重建.结果表明:4种多盘科吸虫为多盘虫属Polystoma和双睾虫属Diplorchis的4个分类单元,其中2未定种为多盘虫属未记录种;肠管、生殖系统、后吸器、大钩和生殖棘等为多盘虫属属内种定种的可靠性状.基于18S rRNA基因序列,4种多盘科吸虫呈现以下进化关系:1)寄生于昭觉林蛙和华西雨蛙的多盘虫属2未定种进入欧非混合进化支;2)锯腿树蛙多盘吸虫位于多盘虫属进化支的最基部;3)石林双睾吸虫进入澳洲进化支.     

Ontogenesis and Molecular Phylogeny of the Marine Ciliate Diophryopsis hystrix: Implications for the Systematics of the Diophrys-like Species (Ciliophora, Spirotrichea, Euplotida)

ABSTRACT. The cortical development of Diophryopsis hystrix , a small marine euplotid ciliate, was observed during binary division. The pattern of morphogenesis is generally similar to that of other Diophrys- like species, with the exception of the marginal cirrus, which develops in a unique manner in D. hystrix being formed from the posterior end of the dorsal kinety anlage, in a similar manner to the formation of the caudal cirri. In the small subunit (SS) rRNA gene sequence tree, Diophryopsis represents a distinct clade separate from Diophrys . Furthermore, our ontogenetic and SSrRNA gene sequence data both support the suggestion that the Diophrys complex is sister to Uronychia , and should be considered as a distinct subfamily within the family Uronychiidae, i.e. Diophryinae Jankowski, 1979, comprising Diophrys , Diophryopsis , and Paradiophrys . Diagnoses of the subfamilies Diophryinae and Uronychiinae are provided.     

Characterization of halotolerant rhizobia isolated from root nodules of Canavalia rosea from seaside areas

Twelve nodule isolates from Canavalia rosea, an indigenous leguminous halophyte growing in the seaside areas of southern Taiwan, were effective symbionts for the original host and able to grow at NaCl concentrations up to 3-3.5% (w/v). The taxonomy of these isolates was investigated using a polyphasic approach, including phenotypic characteristics, banding patterns of total proteins from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), genomic fingerprint patterns from random amplified polymorphic DNA (RAPD) analysis, pulsed-field gel electrophoresis (PFGE) analysis, amplified 16S rDNA restriction analysis (ARDRA), 16S rRNA gene sequencing, and nifH gene sequencing. Based on the SDS-PAGE, RAPD, PFGE and ARDRA results, the 12 isolates are highly diverse. The 16S rRNA and nifH gene sequences were determined for isolates with distinct ARDRA patterns and compared with other members of the rhizobial species. We propose these isolates should be classified into the genus Sinorhizobium and distinguished from the current species of this genus.     

Expanding character sampling for the molecular phylogeny of euplotid ciliates (Protozoa, Ciliophora) using three markers, with a focus on the family Uronychiidae

Although euplotida ciliates are widely used as model organisms in multiple fields of biology, details of their phylogenetic relationships remain unresolved despite a rich history of investigation with small subunit (SSU) rDNA sequences and other characters. Here, six genera in Diophrys-like complex and three other euplotid genera are sampled for SSU-rDNA, ITS1-5.8S-ITS2 and LSU-rDNA, and their phylogenies were inferred with unconstrained and constrained analyses. In general, the concatenated analyses infer more reliable, less ambiguous phylogenies with higher node support values. The following conclusions can be made: (1) four well-supported clades are consistently detected in the family Uronychiidae, forming into two subgroups, which challenge the traditional arrangement based on morphological similarities; (2) the subfamily Diophryinae is paraphyletic; (3) the monophyly of Paradiophrys and the establishment of Apodiophrys and Diophryopsis is fully supported by concatenated data; (4) Apodiophrys and Paradiophrys form independent lineages, at the subfamily level, from other Diophrys-like genera; and (5) the highly specialized Pseudodiophrys nests within Diophrys.     

Rapid identification of marine bioluminescent bacteria by amplified 16S ribosomal RNA gene restriction analysis

To rapidly identify natural isolates of marine bioluminescent bacteria, we developed amplified ribosomal DNA restriction analysis (ARDRA) methods. ARDRA, which is based on the restriction patterns of 16S rRNA gene digested with five enzymes (EcoRI, DdeI, HhaI, HinfI, RsaI), clearly distinguished the 14 species of marine bioluminescent bacteria currently known, which belong to the genera Vibrio, Photobacterium, and Shewanella. When we applied ARDRA to 129 natural isolates from two cruises in Sagami Bay, Japan, 127 were grouped into six ARDRA types with distinctive restriction patterns; these isolates represented the bioluminescent species, P. angustum, P. leiognathi, P. phosphoreum, S. woodyi, V. fischeri, and V. harveyi. The other two isolates showing unexpected ARDRA patterns turned out to have 16S rRNA gene sequences similar to P. leiognathi and P. phosphoreum. Nevertheless, ARDRA provides a simple and fairly robust means for rapid identification of the natural isolates of marine bioluminescent bacteria, and is therefore useful in studying their diversity.     

四核舍太虫的细胞发生学与Helix E10-1二级结构分析

利用蛋白银染色技术,观察和研究海洋游仆虫-四核舍太虫Certesia quadrinucleata(纤毛门,游仆目)二分裂期间的形态发生学。其主要特征如下:(1)老口围带完全被前仔虫继承;(2)后仔虫口原基独立产生于皮膜深层;(3)老口侧膜参与前仔虫口侧膜原基形成,前后仔虫的口侧膜原基均发生于细胞表面, 向前贡献出第一根额腹棘毛;(4)额-腹-横棘毛以初级5原基模式产生, 且以"3:3:3:3:3"的方式分化出新的棘毛;(5)背触毛与左缘棘毛原基均来自老结构, 无尾棘毛产生。研究首次给出了背面纤毛器的发生图示,为进一步探讨舍太虫的系统地位提供了一份补足性的发生学基础资料。游仆目纤毛虫的核糖体小亚基基因HelixE10-1区域二级结构一共存在9种模式, 该区域序列长度的变异性揭示了游仆目纤毛虫在进化中可能处于比较特殊的地位。       

Biodiversity of cultivable psychrotrophic marine bacteria isolated from Terra Nova Bay (Ross Sea, Antarctica)

A set of 146 Antarctic marine isolates from the Ross Sea was characterized by a combination of molecular techniques in order to determine the degree of inter- and intraspecific variability. Isolates were analyzed by amplified rDNA restriction analysis (ARDRA) using the tetrameric enzyme AluI, resulting in 52 different groups, corresponding to at least 52 different bacterial species, indicating a high degree of interspecific variability. The phylogenetic position of bacteria belonging to some ARDRA groups was obtained by sequencing of 16S rDNA. Random amplified polymorphic DNA (RAPD) analysis, carried out on the largest ARDRA groups, revealed a high intraspecific genetic variability, too. The analysis of plasmid content revealed the existence of horizontal gene transfer between strains belonging to the same and to different species. A comparison of the whole body of morphological, physiological and biochemical data was finally carried out.     

Redefinition of the Genera Diophrys and Paradiophrys and Establishment of the Genus Diophryopsis N. G. (Ciliophora, Hypotrichida): Implication for the Species Problem

ABSTRACT. Species belonging to the genus Diophrys Dujardin, 1841, are easily recognized due to possession of the usual complement of approximately seven frontoventral cirri, five transverse cirri, two left marginal cirri, and three large caudal cirri. Separation of these species has been based upon differences in cell length and width, the number and arrangement of cilia in dorsal kinetics, the configuration of the adoral zone of oral polykinetids, the number and distribution of cirri within cirral groups, and the number and arrangement of macronuclei. Jankowski used some of these characteristics to divide the genus into two genera, Diophrys and Paradiophrys, with several subgenera [Jankowski, A. W. 1978. Systematic revision of the class Polyhymenophora (Spirotricha), morphology, systematics and evolution. Tezisy Dokl. Zool. Inst. Akad. Nauk SSSR, 197839-40. (in Russian); Jankowski, A. W. 1979. Systematics and phylogeny of the order Hypotrichida Stein, 1859 (Protozoa, Ciliophora). Trudy. Zool. Inst. Akad. Nauk SSSR, 86 :48–85. (in Russian with English summary)]. Data obtained from light microscopic examination of stained (nigrosin-butanol, Chatton-Lwoff, and Protargol) cells in interphase or division supports and modifies the use of particular structural features of these ciliates for the purpose of taxonomic classification. The structural variability within and among populations of different species within the genera Diophrys (D. appendiculata, D. oligothrix, and D. scutum) and Paradiophrys (P. irmgard and P. multinucleata) is described. D. hystrix is redescribed as the type of the new genus Diophryopsis n. g. Comparative information on the cortical morphogenesis of division of selected species within each genus is reviewed. Two taxonomic classifications of these hypotrichs are discussed: 1) a listing of diagnoses and synonymies and 2) a binary key for identification of all species at the light microscope level. An alternative evolutionary explanation of variations among isolates is presented.     

Molecular characterization of myxozoan parasites from Lake Sasajewun, Algonquin Park, Ontario, by riboprinting

The small subunit-rRNA genes of 18 myxozoans from Lake Sasajewun, Algonquin Park were amplified and digested with restriction endonucleases for riboprinting analysis. Identical riboprints were not found between the myxosporeans and the actinosporeans. The distinct riboprinting patterns observed among these myxozoans indicate considerable genetic diversity within this group. Identical riboprints were found between Myxobolus pendula and Myxobolus pellicides, and between triactinomyxon 'C' and Triactinomyxon ignotum. Parsimony analysis of the riboprints demonstrated that neither the myxosporeans nor the actinosporeans formed a monophyletic group. Some species of Myxobolus are more closely related to forms of triactinomyxon, echinactinomyxon or raabeia than to other Myxobolus species. These results are consistent with the two-host life cycle hypothesis of myxozoans that myxosporeans and actinosporeans are alternating stages of the same organisms.