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。     

海洋纤毛虫黄色伪角毛虫无性生殖期间的形态发生

利用蛋白银染色技术研究了海洋纤毛虫———黄色伪角毛虫Pseudokeronopsisflava (Cohn ,186 6 )Wirns berger,Larsen&Uhlig ,1987无性生殖期间的细胞发生学。其主要特征为 :1)前仔虫口原基以独立发生的方式出现并独特地形成于口前庭右侧的皮层深处 ,由其对老口围带进行完全的更新 ;2 )老口器不参与新口器的形成 ,完全被吸收 ;3)前仔虫的额 -腹 -横棘毛原基同样为独立发生 ,老结构可能不参加其随后的发育 ;4 )后仔虫的口原基、波动膜原基及额 -腹 -横棘毛原基均来自最初排列无序的毛基粒发生场 ;5 )背触毛及缘棘毛的更新发生在老的结构中 ,并向前后延伸取代老结构 ;6 )在整个发生过程中 ,无大核融合现象。文中同时对该种所表现的发生学特征及系统学意义做了探讨     

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。       

急纤虫Tachysoma pellionella无性生殖周期中核器和纤毛器的演化

应用能同时显示纤毛虫的皮膜结构及核器的蛋白银染色方法,研究了急纤虫Tachysoma pellionella的形态及其无性生殖周期中核器和纤毛器的发育演化过程。其中的形态发生过程是:(1)大核改组带出现后,在口围带(AZM)和腹棘毛VC4、VC5之间形成一条细线,于细线中发生许多成群的毛基体,逐渐演变成为后AZM原基区。最终,原基区颗粒组装成一片片整齐排列的小膜,它们构成为新AZM。老AZM也伴随着由基部向前更新,(2)在额棘毛FC5-FC8和腹棘毛VCl-VC3全部瓦解时紧接着产生前、后波动膜原基和新棘毛原基区,并发生波动膜的分化,棘毛的分化、移动和定位过程;(3)左、右缘棘毛原基的发育形式相同,但右缘棘毛原基的发育稍早。在右缘棘毛列中的第2(或第3)根和约第16根棘毛、左缘棘毛列的第1根和约第17根棘毛开始,随老棘毛基部的瓦解,于老棘毛基部位置各产生前、后两部分新缘棘毛原基,左、右每部分原基各占据了约六个老棘毛基部位置后分别朝所在的老棘毛列的外侧、内侧向后伸展开来;(4)在第1—3列背触毛中分别于每列之前、后两部分的中部范围产生前、后第1—3列新原基,每列原基向其两端伸展替代老背触毛列,它们后来成为前、后仔虫的相应的第1—3列新背触毛。接着在前,后各第3列背触毛原基后端发生前、后第4列原基,并稍偏向第3列原     

红色角毛虫生理改组过程的研究

红色角毛虫在生理改组时,随着老纤毛器的瓦解,先后出现新的口器,额、腹、横棘毛,左、右缘棘毛和背触毛四个原基区,并发生原基区的分化、新结构的形成和定位。这种新、老结构的更替过程相似于同种纤毛虫正常形态发生时期纤毛器的演化过程,口围带改组时,新口围带原基在左列中腹棘毛左侧的范围形成,后来,随着老口围带的瓦解,它向前方移动并处于老口围带的右侧,并继续朝老口围带位置移动、替换老口围带。这不同于其他常见的腹毛类纤毛虫,生理改组时新口围带原基在瓦解着的老口围带的位置逐渐移动替换老口围带的情况。     

拟伍氏游仆虫(原生动物,纤毛门,游仆目)的重描述

利用活体观察和银染技术,对采自广东省惠阳大亚湾沿岸的一种罕见海洋游仆类纤毛虫拟伍氏游仆虫Euplotes parawoodruffi进行了再研究,补充了有关活体形态学、性状变异以及分类学特征的统计学资料等。中国新纪录种拟伍氏游仆虫大亚湾种群与原报道种群存在细微差异,其主要特征为:口围带小膜数为60~70;体纤毛器数目及排布模式十分稳定:额腹棘毛9根,横棘毛5根,尾棘毛2根,左缘棘毛2根;背触毛恒为9列;大核近T形,形状多变。     

阔口尖毛虫皮层纤毛器微管在不同生理条件下的分化

应用激光扫描共聚焦显微术,显示腹毛类纤毛虫阔口尖毛虫(Oxytricha platystoma)无性生殖过程中,新的口围带、波动膜、额腹横棘毛、左右缘棘毛微管先后分化,老纤毛器微管去分化,细胞分裂产生各含一套纤毛器微管的前、后两仔虫;生理改组过程中,口围带、波动膜、额腹横棘毛、左右缘棘毛微管发生去分化和再分化,细胞皮层微管胞器更新形成含一套纤毛器微管的新细胞。结果表明阔口尖毛虫在无性生殖和生理改组这两种不同的生理条件下,其纤毛器微管结构的形成或更新可能具有相同的细胞调控机制,形态发生中老纤毛器结构可能对新结构的发生和发育具有诱导定位和物质贡献的作用。     

悬游双眉虫(原生动物,纤毛门)无性生殖期间的形态发生

应用蛋白银染色技术研究了悬游双眉虫青岛种群无性生殖期间皮层结构和核器的演化过程,其主要特征为:后仔虫口原基独立地出现于紧靠虫体左侧第一根横棘毛的皮层下小龛,其中毛基粒不参与其它棘毛原基的形成;老的口围带发生后半部的局部重建而非整个的由前仔虫简单继承;在前仔虫中,波动膜原基来自老结构的反分化,而在后仔虫中则来自口原基;所有棘毛原基均为独立发生并与老结构没有任何关系;在前仔虫中,口棘毛(即左侧第一根额棘毛)来自于波动膜的反分化,而在后仔虫中则为独立发生;背触毛列于老的结构当中产生,并由最右一列原基演化出3根尾棘毛;两大核片段的改组带从一端向另一端移动 ,并随着两者的融合而消失.文中同时对前人有关该属发生模式的若干存疑问题做了探讨 [动物学报 54(3):517-524,2008].     

阔口尖毛虫无性生殖和生理改组过程的比较

阔口尖毛虫无性生殖中先后发生后口围带原基,前、后的波动膜原基,额腹横棘毛原基和左、右缘棘毛原基,老口围带也在此期间更新,结果形成2套新纤毛结构,原老结构瓦解消失;生理改组时按同样顺序产生口围带原基等几类腹面纤毛原基,结果形成1套新纤毛结构,替换老结构。在这两个截然不同的过程中,新纤毛结构的分化和老纤毛结构退化时也表现出某些相似的特征。作者据此推测,这种纤毛虫无性生殖和生理改组中,纤毛原基的发生、发育和定位在细胞控制机理上可能是相同的。     

多核织毛虫─新种,无性生殖周期中的形态及形态发生

本新种的典型特征为:表膜较薄但不弯曲,无表膜下颗粒;区别于该属其它种的最突出特点是具有多个大核(5.4±1.4);背触毛6列,在第3-4列之间具有一些额外散在的背触毛.口围由42.3±2.2片小膜组成;额、腹、横、尾棘毛恒为:5:5:3结构;本新种的形态发生结果与该属其它种类相同,6列新背触毛原基分别来源于背部的3列老背触毛和腹部的老右缘棘毛,在第3及第4列新背触毛列之间可见新出现并散在的几根背触毛.       

The morphology and morphogenesis of a marine ciliate,Epiclintes auricularis rarisetus nov. sspec. (Ciliophora,Epiclintidae), from the Yellow Sea

The morphology and morphogenesis of a marine ciliate, Epiclintes auricularis rarisetus nov. sspec., collected from Qingdao, northern China were studied on live and protargol-impregnated specimens. This isolate can be recognized by its elongate, contractile and tripartite body with a size of 200-400×20-40 μm in vivo, about 30 ventrally located adoral membranelles, short undulating membranes, 2-3 frontal and 10-18 transverse cirri as well as 8-9 fronto-midventral rows, 22-31 left and 35-54 right marginal cirri, but no caudal cirri, invariably 3 dorsal kineties, of which kineties 1 and 3 link together anteriorly, 24-70 macronuclear nodules, marine habitat, and contribution of almost all frontal-midventral transverse cirral anlagen (FVT anlagen) to the formation of fronto-midventral rows. In its divisional morphogenesis, this species demonstrates some features rarely if ever found in any other urostyloids, e.g. partial replacement of the old adoral zone of membranelles, de novo formation of the oral primordium and the anlagen for marginal rows and dorsal kineties, contribution of almost all FVT anlagen to the transverse cirri, lack of frontoterminal cirri. In addition, its morphogenesis appears to differ in detail from that of E. auricularis auricularis nov. stat., based on published data. These remarkable morphogenetic traits of Epiclintes suggests a questionable systematic position for this genus in the Urostyloidea, therefore, further researches are urgently needed.     

Morphology,ontogeny and molecular phylogeny of a new urostylid ciliate,Bakuella (Pseudobakuella) guangdongica n. sp. (Ciliophora,Hypotrichia) from southern China

The morphology, morphogenesis, and molecular phylogeny of Bakuella (Pseudobakuella) guangdongica n. sp., isolated from southern China, were investigated. The new species is characterized by a body length of 150–225 μm in vivo; 35–42 adoral membranelles; 3–5 buccal, two frontoterminal, 7–12 transverse and two pretransverse ventral cirri; midventral complex comprised of 10–20 pairs and two rows extending to transverse cirri; posterior part of marginal rows slightly overlapping; colorless cortical granules about 1 μm across, arranged in small groups; soil habitat. Its main ontogenetic features are: (1) in the proter, the parental adoral zone of membranelles is completely renewed by new structures; (2) in the opisthe, the oral primordium originates apokinetally, some old midventral cirri join the formation of frontoventral-transverse cirral anlagen; (3) the anlagen for marginal rows and dorsal kineties develop intrakinetally; and (4) the numerous macronuclear nodules fuse into a single mass before dividing. Phylogenetic analyses based on the SSU rDNA sequence suggest the non-monophyly of the genus Bakuella.     

Cortical structure in non-dividing and dividing Diophrys japonica spec. nov. (Ciliophora, Euplotida) with notes on morphological variation

The morphology and morphogenesis of Diophrys japonica spec. nov., isolated from the Mie Port, Nagasaki, Japan, were investigated from life and following impregnation with protargol. The new species is recognized by the following characters: Body elliptical in outline and slightly greyish to yellowish in color; size in vivo about 80-120 x 50-70 microm; pellicle flexible, with underlying granules densely arranged in lines; ciliature comprising about 30-46 adoral membranelles, 4-7 frontal, 1-4 ventral and 4-7 transverse cirri, always 1 left marginal and 3 caudal cirri, and 4 dorsal kineties; usually two macronuclear nodules; fragment kinety with 2-5 dikinetids; marine habitat. The main morphogenetic events are: (1) the opisthe's oral primordium develops de novo in a subsurface pouch near the left transverse cirri; (2) the proter retains the parental AZM except for reorganization of some proximal membranelles; (3) cirral anlagen for the frontal, ventral and transverse cirri in both dividers develop separately from the oral primordium or parental cirri, and are derived from the separation of primary primordia that originate de novo; (4) the anlagen for the left marginal cirrus and fragment kinety also form de novo and separately; (5) dorsal kinety anlagen occur within the parental structures at mid-body and posterior end of the cell, of which the right-most one contributes three caudal cirri from its posterior portion. Based on available ontogenetic data, the author proposes that the numbers of left marginal and caudal cirri can be regarded as reliable characters for species identification, while the numbers of frontal, ventral and transverse cirri are not consistent enough for species distinction. A key to the eleven adequately known species of Diophrys is presented.     

Morphogenesis in the marine spirotrichous ciliate Apokeronopsis crassa (Claparède & Lachmann, 1858) n. comb. (Ciliophora: Stichotrichia), with the establishment of a new genus, Apokeronopsis n. g., and redefinition of the genus Thigmokeronopsis

Morphogenetic events during the division of the marine spirotrichous ciliate, Apokeronopsis crassa (Claparède & Lachmann 1858) n. comb. were investigated. Compared with members of the well-known genera Thigmokeronopsis, Uroleptopsis, and Pseudokeronopsis, A. crassa has one row of buccal cirri, high number of transverse cirri, clearly separated midventral rows, lacks thigmotactic cirri and a gap in adoral zone, its undulating membranes (UMs) anlage forms one cirrus and marginal rows and dorsal kineties form apokinetally during division. All these characteristics indicate that this organism represents a new taxon at the generic level, and hence a new genus is suggested, Apokeronopsis n. g. It is defined as thus: Pseudokeronopsidae with Pseudokeronopsis-like bicorona of frontal cirri and one marginal row on each side; one row of two or more buccal cirri in ordinary position; two midventral rows distinctly separated, hence of cirri that are not in a typical zig-zag pattern; high number of transverse cirri, caudal cirri absent, and frontoterminal cirri present; thigmotactic cirri absent, many macronuclear nodules fuse into many masses as well as marginal and dorsal kineties form apokinetally during morphogenesis. At the same time, the genus ThigmokeronopsisWicklow, 1981 is redefined, and one new combination, Apokeronopsis antarctica (Petz, 1995) n. comb. is proposed. The morphogenetic events of A. crassa are characterized as follows: (1) In the proter, the adoral zone of membranelles and UMs are completely renewed by the oral primordium. The UM anlage is formed apokinetally on the dorsal wall of the buccal cavity and is hence clearly separated from the frontoventral-transverse (FVT) cirral anlagen in the proter. (2) Frontoventral-transverse cirral anlagen are generated de novo in the outermost region of the cortex to the right of the old UMs. (3) A row of buccal cirri arises from FVT cirral streak I. (4) The marginal rows and dorsal kineties originate de novo in both dividers; no caudal cirri are formed. (5) The last FVT-streak contributes two frontoterminal cirri. (6) The many macronuclear nodules fuse into many masses (about 50 segments) during division, unlike a singular or branched mass as described in other urostylids.     

Morphology and cell division of Saudithrix terricola n. gen., n. sp., a large, stichotrich ciliate from Saudi Arabia

The morphology and main ontogenetic traits of a new stichotrich ciliate, Saudithrix terricola Foissner, AL-Rasheid and Berger n. gen., n. sp., from a terrestrial habitat in Saudi Arabia were investigated using live observation, protargol impregnation, and scanning electron microscopy. Saudithrix terricola is characterized by a large (200-350 x 70-150 microm), flexible body; an adoral zone formed like a three-quarter circle; a sickle-shaped buccal lip with a widened paroral forming a cyrtohymenid pattern with the endoral; 11 frontal and frontal-ventral cirral rows (including right marginal rows) and one left marginal row covering the ventral side; two buccal cirri; six to nine transverse cirri; three dorsal kineties; and two macronuclear nodules. The resting cyst is about 85 microm across, has a smooth wall, and a fluffy mucous layer. Most cirral anlagen originate within the parental rows and are arranged side by side, the proximal portion of the adoral zone of membranelles is reorganized, and some parental dorsal bristles are maintained. Neither the morphological nor the ontogenetic data reveal the systematic position of Saudithrix within the stichotrichs. The term multicorona is introduced and describes a frontal ciliature composed of four or more cirral bows.     

Morphology,morphogenesis and molecular phylogeny of a novel soil ciliate,Afrokahliella paramacrostoma n. sp. (Ciliophora,Hypotrichia)

A soil hypotrich ciliate, Afrokahliella paramacrostoma n. sp., was discovered in China. Its morphology, morphogenesis and molecular phylogeny were investigated using standard methods. The new species is characterized as follows: body about 140–180 × 60–70 μm in vivo, cortical granules absent, contractile vacuole positioned about 40% down length of body, 5–9 macronuclear nodules, 34–49 adoral membranelles, 3–5 buccal and 3–6 parabuccal cirri, usually two frontoventral rows, three or four left and two or three right marginal rows, three dorsal kineties and one dorsomarginal kinety; 1–3 and one or two caudal cirri located at the ends of dorsal kineties 1 and 2, respectively. The ontogenetic process is characterized by: (1) the marginal anlagen on each side develop in the outer right and the inner left marginal rows, respectively; (2) five frontoventral-transverse cirral anlagen, anlagen II–IV develop in secondary mode; (3) dorsal morphogenesis follows a typical Urosomoida-pattern, no parental dorsal kineties are retained; (4) caudal cirri are generated at the ends of dorsal kineties 1 and 2. Phylogenetic analyses based on SSU rDNA sequence data reveals that Afrokahliella paramacrostoma n. sp. is closely related to Parakahliella macrostoma and Hemiurosomoida longa.     

Morphology and morphogenesis of Holosticha heterofoissneri nov. spec. from the Yellow Sea,China (Ciliophora,Hypotrichida)

The morphology and morphogenesis of the marine hypotrichous ciliate Holosticha heterofoissneri nov. spec. from mollusc culture in Qingdao, China are described based on living and protargol-impregnated specimens. The new species is characterized by: adoral zone slightly bipartite, consisting of on average 43 membranelles; constantly 5 dorsal kineties, and 12 pairs of cirri in midventral rows, and 14–16 macronuclear segments, which differs from the closely related species, H. foissneri. Its morphogenetic process shows the following features: (1) the parental AZM is retained unchanged and will be inherited by the proter, only the old UM reorganized; (2) FVT-cirral anlagen in both division parts derive from the breaking of primary primordia; (3) during the formation of FVT-cirral anlagen, most midventral cirri remain intact; and there appears an 'extra' anlage (EA) between the UMA and other FVT-cirral primordia; (4) the generation type of the dorsal kineties is of 'one group mode', the leftmost one of the 4 primordia fragmentates to form a new kinety; (5) at the early stage of morphogenesis, replication bands of macronuclear segments are apparently present.     

Morphology and infraciliature of three species of Metaurostylopsis (Ciliophora, Stichotrichia): M. songi n. sp., M. salina n. sp., and M. marina (Kahl 1932) from sediments, saline ponds, and coastal waters

Two new urostylid ciliates, Metaurostylopsis songi n. sp. and Metaurostylopsis salina n. sp. and Metaurostylopsis marina (Kahl 1932) are investigated using live observation and protargol impregnation. These species were isolated in Korea from intertidal sediments, saline ponds, and coastal waters. Metaurostylopsis songi is in vivo about 120 microm x 25 microm, has a slenderly ellipsoidal body, colorless cortical granules in rows on ventral and dorsal body sides, about 54 macronuclear nodules, 28-47 adoral membranelles, five frontal, two or three frontoterminal and six or seven transverse cirri, and 9-12 midventral cirral pairs followed posteriorly by 1-3 single cirri. In vivo M. salina is about 60 microm x 25 microm, has a pyriform body, colorless cortical granules irregularly arranged, about 45 macronuclear nodules, 18-23 adoral membranelles, three frontal, three to five frontoterminal and two to five transverse cirri, and four or five midventral cirral pairs followed posteriorly by five to seven single cirri. Both species have three marginal cirral rows on each body side and 3 long dorsal kineties. The Korean specimens of M. marina match the Chinese population in all main features. Metaurostylopsis songi differs from M. marina by the more slender body, the number of frontal cirri (invariably five vs. four), and the arrangement of cortical granules (in rows on dorsal and ventral cortex vs. only along dorsal kineties and anterior body margin). Metaurostylopsis salina differs from its congeners by the distinctly smaller size, the pyriform body shape, the scattered cortical granules (vs. in rows), and number of frontal cirri. It differs from M. marina also by the number of midventral cirral pairs (four or five vs. seven to 11).     

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。       

Morphogenesis of the freshwater ciliate Neokeronopsis spectabilis (Kahl 1932) Warren et al., 2002, based on a China population (Ciliophora: Urostylidae)

The morphogenesis of the stichotrichous ciliate, Neokeronopsis spectabilis, collected from a freshwater pond near Harbin, north China, was observed following protargol impregnation. The overall morphogenetic events are characterized by: (1) the new oral primordium originates in association with the transverse cirri; (2) the proter's undulating membrane anlage is formed following the dedifferentiation of the parental endoral and paroral membranes, while the old adoral zone of membranelles is retained; (3) the fronto-ventral-transverse cirral anlagen originate independently on the right of the proter's undulating membrane anlage; (4) the left and right marginal cirri are derived from new anlagen that originate within the old marginal rows; (5) the generation of the dorsal kineties is of the "two-group-mode" with fragmentation and hence an oxytrichid pattern; and (6) similar to that in oxytrichids, the caudal cirri are formed at the posterior ends of the rightmost and two leftmost kineties of the group 1. Some new morphological data for N. spectabilis are also presented and the systematic position of the genus Neokeronopsis is briefly discussed. We conclude that its position among the urostylids is peripheral and that it very likely represents an intermediate form between the oxytrichids and urostylids.