RECONSTRUCTION OF THE FEEDING APPARATUS IN PLOEOTIA COSTATA (EUGLENOPHYTA) AND ITS RELATIONSHIP TO OTHER EUGLENOID FEEDING APPARATUSES

The ultrastructure of the feeding apparatus in Ploeotia costata Farmer and Triemer was determined and compared to other euglenoid feeding apparatuses. The feeding apparatus opened subapically onto the ventral surface and extended nearly the entire length of the cell. It consisted of four parts at the anterior surface: a comb, cytostome/pocket, vanes, and supporting rods. The comb was a multilayered structure of three horizontal microtubular rows encased in cement and formed the dorsal lip of the apparatus. The cytostome/pocket was located between the comb and the supporting rods, tapered into the cell as the cytopharynx and was surrounded by five vanes. The electron-opaque vanes extended the entire length of the feeding apparatus and were lined with microtubules for most of their length. Finally, two cement supporting rods that were joined by a crosspiece at the anterior end formed the ventral lip. The rods separated briefly before merging with the vanes. As the merged rods and vanes descended into the cell, they gradually narrowed and terminated. Comparisons of the feeding apparatus with Ploeotia vitrea, Diplonema ambulator, Lentomonas applanatum, and other euglenoids have led to the conclusion that the Type II feeding apparatus is found only in Ploeotia species.     

An Ultrastructural Study of Lentomonas applanatum (Preisig) N. G. (Euglenida)

ABSTRACT. Lentomonas applanatum (syn. Entosiphon applanatum Preisig) is a biflagellate, phagotrophic euglenid found in intertidal salt marshes. Lentomonas applanatum bears a superficial similarity to Entosiphon sulcatum , however, an ultrastructural study of L. applanatum revealed many features that are atypical for other described species of the genus Entosiphon . These features include number and organization of pellicular strips, construction of the feeding apparatus, nature of the flagellar transition zone and flagellar apparatus, and point of flagellar emergence. These differences show that L. applanatum is related more closely to phagotrophic genera such as Ploeotia than to E. sulcatum . The construction of the feeding apparatus and pellicle suggest that L. applanatum has retained many of the pleisiomorphic characters that were present in the earliest euglenids. The presence of similar structures in other related protists may provide important clues as to the evolution of the Euglenida.     

Discovery of a group I intron in the SSU rDNA of Ploeotia costata (Euglenozoa)

The gene coding for the small ribosomal subunit RNA of Ploeotia costata contains an actively splicing group I intron (Pco.S516) which is unique among euglenozoans. Secondary structure predictions indicate that paired segments P1-P10 as well as several conserved elements typical of group I introns and of subclass IC1 in particular are present. Phylogenetic analyses of SSU rDNA sequences demonstrate a well-supported placement of Ploeotia costata within the Euglenozoa; whereas, analyses of intron data sets uncover a close phylogenetic relation of Pco.S516 to S-516 introns from Acanthamoeba, Aureoumbra lagunensis (Stramenopila) and red algae of the order Bangiales. Discrepancies between SSU rDNA and intron phylogenies suggest horizontal spread of the group I intron. Monophyly of IC1 516 introns from Ploeotia costata, A. lagunensis and rhodophytes is supported by a unique secondary structure element: helix P5b possesses an insertion of 19 nt length with a highly conserved tetraloop which is supposed to take part in tertiary interactions. Neither functional nor degenerated ORFs coding for homing endonucleases can be identified in Pco.S516. Nevertheless, degenerated ORFs with His-Cys box motifs in closely related intron sequences indicate that homing may have occurred during evolution of the investigated intron group.     

Strains of the Morphospecies Ploeotia costata (Euglenozoa) Isolated from the Western North Pacific (Taiwan) Reveal Substantial Genetic Differences

Two phagotrophic euglenid strains (Strains Pac and Tam) were isolated from coastal locations in Taiwan. Ultrastructural characteristics of the strains included five pellicle strips joined at the posterior end. The strips were formed by major grooves with bifurcated edges. At the cell anterior, the feeding structure formed a lip. Underneath the lip was a comb composed of layers of microtubules. Farther back, two supporting rods tapered toward the posterior end, and a number of vanes with attached microtubules were present between the rods. The morphological characteristics agree with Ploeotia costata Strain CCAP 1265/1. However, the 18S rDNA sequences of Strains Pac/Tam lacked a group I intron and possessed three extra insertions of 116, 67, and 53 bp. Phylogenetic analysis indicated low sequence similarity between Strains Pac/Tam and CCAP 1265/1 (92%). The morphospecies P. costata apparently includes a substantial level of DNA sequence divergence, and likely represents multiple molecular species units.     

The flagellar apparatus of spermatozoa in fish. Ultrastructure and evolution

Chondrichthyes possess an evolved type of spermatozoa. Their flagellar apparatus is characterized by the presence of flagellar roots which form the axis of the midpiece, and the existence of one or two lateral elements associated with the axoneme. Osteichthyes, mainly teleosteans, show a great diversity of spermatic forms. The primitive spermatozoon with a 9 + 2 pattern flagellum is common. The primitive spermatozoon has evolved along different lines. The spermatic diversity which results from this is mainly evident in the structure of the flagellar apparatus. In the animal kingdom the primitive spermatozoon with a 9 + 2 pattern flagellum, present in primitive metazoa, is retained in phyla where external fertilization is maintained. The main evolutionary tendencies--elongation, aflagellarity or biflagellarity--are generally connected with the acquisition of internal fertilization. These evolutionary tendencies are found in teleosteans. It is not possible to link aflagellarity or biflagellarity of the gamete in certain fishes to this method of fertilization. Only the elongation of the spermatozoon is connected, in certain cases, with internal fertilization, but this cannot be taken as general.     

Reconstruction of the feeding apparatus in Postgaardi mariagerensis provides evidence for character evolution within the Symbiontida (Euglenozoa)

Microbial eukaryotes living in low oxygen environments often have novel physiological and morphological features that facilitate symbiotic relationships with bacteria and other means for acquiring nutrients. Comparative studies of these features provide evidence for phylogenetic relationships and evolutionary history. Postgaardi mariagerensis, for instance, is a euglenozoan that lives in low oxygen environments and is enveloped by episymbiotic bacteria. The general ultrastructure of P. mariagerensis was described more than a decade ago and no further studies have been carried out since, mainly because these cells are difficult to obtain. Postgaardi lacks the diagnostic features found in other major euglenozoan lineages (e.g., pellicle strips and kinetoplast-like mitochondrial inclusions) and no molecular data are available, so the phylogenetic position of this genus within the Euglenozoa remains unclear. We re-examined and reconstructed the ultrastructural organization of the feeding apparatus in Postgaardi by serial sectioning an existing block of resin-embedded cells. Postgaardi possesses distinctive finger-like projections within the feeding apparatus; this system has only been found in one other highly distinctive flagellate, namely the symbiontid Calkinsia. Detailed comparisons of the cytoskeleton in Postgaardi and in two symbiontids, Calkinsia and Bihospites, provided new evidence for phylogenetic relationships and character evolution in all three genera.     

Reconstruction of the flagellar apparatus and microtubular cytoskeleton inPyramimonas gelidicola (Prasinophyceae,Chlorophyta)

Summary The absolute configuration of the flagellar apparatus inPyramimonas gelidicola McFadden et al. has been determined and shows identity withP. obovata, indicating that they are closely related. Comparison with the flagellar apparatus of quadriflagellate zoospores from the more advancedChlorophyceae suggest thatPyramimonas may be a primitive ancestral form. The microtubular cytoskeleton has been examined in detail and is shown to be unusual in that it does not attach to the flagellar apparatus. Cytoskeletal microtubules are nucleated individually, and this is interpreted as an adaptation to the methods of mitosis and scale deployment. In view of the primitive nature of these processes, it is proposed that this type of cytoskeletal organization may represent a less advanced condition than that of the flagellar root MTOCs (microtubule organizing centers) observed in theChlorophyceae.     

The flagellar apparatus structure inMicrospora (Chlorophyceae) confirms a close evolutionary relationship with unicellular green algae

The spatial configuration of the flagellar apparatus of the biflagellate zoospores of the green algal genusMicrospora is reconstructed by serial sectioning analysis using transmission electron microscopy. Along with the unequal length of the flagella, the most remarkable characteristics of the flagellar apparatus are: (1) the subapical emergence of the flagella (especially apparent with scanning electron microscopy); (2) the parallel orientation of the two basal bodies which are interconnected by a prominent one-piece distal connecting fiber; (3) the unique ultrastructure of the distal connecting fiber composed of a central tubular region which is bordered on both sides by a striated zone; (4) the different origin of the d-rootlets from their relative basal bodies; (5) the asymmetry of the papillar region which together with the subapical position of the basal bodies apparently cause the different paths of corresponding rootlets in the zoospore anterior; (6) the presence of single-membered d-rootlets and multi-membered s-rootlets resulting in a 7-1-7-1 cruciate microtubular root system which, through the different rootlet origin, does not exhibit a strict 180° rotational symmetry. It is speculated that the different basal body origin of the d-rootlets is correlated with the subapical implant of flagella. It is further hypothesized that in the course of evolution the ancestors ofMicrospora had a flagellar papilla that has migrated from a strictly apical position towards a subapical position. Simultaneously, ancestral shift of flagella along the apical cell body periphery has taken place as can be concluded from the presence of an upper flagellum overlying a lower flagellum in the flagellar apparatus ofMicrospora. The basic features of the flagellar apparatus of theMicrospora zoospore resemble those of the coccoid green algal generaDictyochloris andBracteacoccus and also those of the flagellate green algal genusHeterochlamydomonas. This strengthens the general supposition thatMicrospora is evolutionarily closely related to taxa which were formerly classified in the traditionalChlorococcales.     

Ultrastructure of the flagellar apparatus inPleurochrysis (classPrymnesiophyceae)

Summary The ultrastructure of the flagellar apparatus of aPleurochrysis, a coccolithophorid was studied in detail. Three major fibrous connecting bands and several accessory fibrous bands link the basal bodies, haptonema and microtubular flagellar roots. The asymmetrical flagellar root system is composed of three different microtubular roots (referred to here as roots 1,2, and 3) and a fibrous root. Root 1, associated with one of the basal bodies, is of the compound type, constructed of two sets of microtubules,viz. a broad sheet consisting of up to twenty closely aligned microtubules, and a secondary bundle made up of 100–200 microtubules which arises at right angles to the former. A thin electron-dense plate occurs on the surface of the microtubular sheet opposite the secondary bundle. The fibrous root arises from the same basal body and passes along the plasmalemma together with the microtubular sheet of root 1. Root 2 is also of the compound type and arises from one of the major connecting bands (called a distal band) as a four-stranded microtubular root and extends in the opposite direction to the haptonema. From this stranded root a secondary bundle of microtubules arises at approximately right angle. Root 3 is a more simple type, composed of at least six microtubules which are associated with the basal body. The flagellar transition region was found to be unusual for the classPrymnesiophyceae. The phylogenetic significance of the flagellar apparatus in thePrymnesiophyceae is discussed.     

女性尿道阴道瘘数字化三维重建在临床治疗中的初步应用

目的:基于CT扫描图象建立精确的女性尿道阴道瘘数字化模型,探讨其在临床诊断及治疗中的应用。方法:选择1例女性尿道阴道瘘病例,进行尿道CT连续断层扫描,扫描结果导入Mimics软件中进行三维重建,利用三维重建模型指导临床。结果:建立女性尿道阴道瘘及周围结构的三维立体模型,可以方便地从任意角度和方向观察瘘管情况,测量有关的数据;还可以在数字化模型上进行手术设计。结论:女性尿道阴道瘘的数字化三维模型能够更直观、准确地反映病变部位的三维立体结构。对女性尿道阴道瘘的诊断、手术规划等有较大帮助。     

Ultrastructure of the flagellar apparatus inPyramimonas octopus (Prasinophyceae)

Summary While green algae usually lack one of the outer dynein arms in the axoneme, flagella of the octoflagellated prasinophytePyramimonas octopus possess dynein arms on all peripheral doublets. The outer dynein arm on doublet no. 1 is modified, and additional structures are associated with doublets no. 2 and 6. The flagellar scales are asymmetrically arranged. Thus the two rows of thick flagellar hairscales are displaced towards doublet no. 6,i.e., in the direction of the effective stroke of each flagellum. The underlayer of small scales includes two nearly opposite double rows scales, arranged in the longitudinal direction of the flagellum. The hairscales emerge from these rows. The double rows are separated on one side by 9, on the other by 11 rows of helically arranged scales. The central pair of microtubules twists, but the axoneme itself (represented by the 9 peripheral doublets), does not seem to rotate. The flagella are arranged in two groups, showing modified 180° rotational symmetry. The effective strokes of the two central flagella are exactly opposite, while the other flagella beat in six intermediate directions.     

The architecture of the flagellar apparatus ofPrymnesium patellifera (Prymnesiophyta)

The flagellar apparatus of the small prymnesiophytePrymnesium patellifera has been analysed and a reconstruction is presented. Externally, the cell carries two sub-equal flagella and a short non-coiling haptonema. Within the cell, there are four microtubular roots and a number of fibrous bands, the latter interconnecting the two basal bodies and the haptonema base. One of the roots (r1) consists of a sheet of up to 25 microtubules originating close to the proximal extremity of the haptonema base, but the other three roots are composed of between 1 and 4 microtubules only. Distally, a large striated fibrous auxiliary connecting root extends across the anterior part of the cell linking root r1 and a mitochondrial profile on the opposite side of the cell. The arrangement of the components of the flagellar apparatus ofP. patellifera is commensurate with the general pattern found in many prymnesiophytes other than members of the Pavlovales, but there are a number of differences in detail from the other species described hitherto.     

The copulatory apparatus of the marine gastropod Haminella solitaria (Heterobranchia: Cephalaspidea) and its phylogenetic relevance

The hermaphroditic marine snail species Haminella solitaria was formerly included in the genus Haminoea, but it was recently assigned to the genus Haminella. The copulatory apparatus in H. solitaria was investigated by light and transmission electron microscopy to obtain additional information about this apparatus in cephalaspidean gastropods and to evaluate the taxonomic relevance of its morphofunctional features in the framework of a new phylogenetic tree of the family Haminoeidae. The copulatory apparatus in H. solitaria consisted of the atrium with a muscular wall and papilla, a seminal duct, and a single‐lobed prostate. Epithelial and subepithelial secretory cells were detected in the proximal and middle region of the atrium wall, and a third type of secretory cell occurred in the distal region of the muscular papilla. The seminal duct was lined by ciliated cells and its muscular wall included some vacuolar cells. The prostate in H. solitaria consisted of lateral pouches surrounding a large central lumen that was filled with spermatozoa. A single type of secretory cell intermingled with ciliated cells formed the epithelium of the prostate. A histological comparison between the copulatory apparatus in H. solitaria and Haminoea navicula revealed substantial differences that support the placement of these two species in different genera, as established by recent molecular studies.     

The ultrastructure of the ascus apical apparatus of some Dermateaceae (Helotiales)

The ascus apical apparatus ultrastructure of 12 species of the Dermateaceae (Helotiales, Ascomycota) was studied. Its development and details are described on the basis of transmission electron microscopy micrographs for all studied species. A majority of them, 8 species belonging to the genera Belonopsis, Mollisia, and Pyrenopeziza, show considerable similarity in the structure of their ascus apical apparatus. Two species, Mollisia melaleuca and Mollisia ramealis, show some deviations from this type, suggesting that the genus Mollisia is not fully homogeneous. Two other species, Podophacidium xanthomelum and Pezicula cinnamomea represent two ascus apical apparatus structure types of their own, which are remarkably different from the Mollisia type. The possible taxonomic implications arising from ascus apical apparatus diversity in the Dermateaceae are discussed.     

Transformation and development of the flagellar apparatus ofCryptomonas ovata (Cryptophyceae) during cell division

Summary InCryptomonas ovata, long, dorsal flagella are produced which transform during the following cell division into short, ventral flagella. At division there is a reorientation in cell polarity, and the parental basal apparatus, which comprises the basal bodies and associated roots, is distributed to the daughter cells via a complex sequence of events. Flagellar apparatus development includes the transformation of a four-stranded microtubular root into a mature root of different structure and function. Each newly formed basal body nucleates new microtubular roots, but receives a striated fibrous root from a parental basal body. The striated roots are originally produced on the transforming basal body and are transferred to the new basal bodies at each successive division. The development of the asymmetric flagellar apparatus throughout the cell cycle is described.     

Observations on the flagellar apparatus of a coccolithophorid,Cruciplacolithus neohelis (Prymnesiophyceae)

The flagellar apparatus ofCruciplacolithus neohelis (McIntyre and Bé) Reinhardt including its transition region is described. The transition region contains a hat-shaped structure, which is suggested to be one of the common features of the Prymnesiophyceae. Its flagellar root system resembles that of most coccolithophorids examined so far, except that only one vestigial crystalline root is present associated with root 1. Two well-developed crystalline roots associated with roots 1 and 2, respectively, appear in the preprophase of nuclear division, suggesting conversion to a mitotic spindle. The taxonomic and evolutionary significance of the flagellar apparatus is discussed.     

Vischeria stellata (Eustigmatophyceae): ultrastructure of the zoospores,with special reference to the flagellar apparatus

Summary Ultrastructure of the zoospores ofVischeria stellata (R. Chodat ex Poulton) Pascher is investigated, with particular reference to the system of flagellar roots. Microtubular roots and a rhizoplast are present and a model showing their distribution is proposed. Four microtubular roots attach to the basal bodies in a system basically similar to that displayed by the heterokont algae and fungi. The rhizoplast is also similar to that of other heterokont algae. We conclude from these observations that the class Eustigmatophyceae should be placed within the division Heterokontophyta.Abbreviations C chloroplast - B basal body of the emergent flagellum - B' second basal body - E eyespot - F emergent flagellum - FS flagellar swelling - LV lamellate vesicle - M mastigonemes - MTs microtubules - N nucleus - R 1–R 4 microtubular roots - Rh rhizoplast - SB striated band - SV spiral vesicle     

Ultrastructure of the dinoflagellate Polykrikos. II. The nucleus and its connections to the flagellar apparatus

Electron microscopy of the colonial dinoflagellate Polykrikos kofoidi revealed a nuclear cortex formed of two electron-dense cortical layers directly beneath the nuclear envelope. Nuclear pores were confined to vesicular outpocketings of the nuclear envelope over circular discontinuities in the cortical layers. A conspicuous fibrous ribbon extended from the nucleus to the flagellar apparatus of each zooid. The ribbons resembled in their structure and position the attractophores of termite flagellates. Each flagellar apparatus consisted of two flagella, two elongate axial kinetosomes, an oblique kinetosome, and two roots of markedly different periodicities.