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.     

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.     

Flagellar regeneration and associated scale deposition inPyramimonas gelidicola (Prasinophyceae,Chlorophyta)

Summary During regeneration of mechanically amputated flagella, flagellar scales and the subtending membrane accumulate in a villiform scale reservoir in which the scales interact to form patterns on the villi reminiscent of the arrangement they later assume on the flagellum. The reservoir membrane is continuous with the plasmalemma, and the scales, attached directly and indirectly to the membrane, leave the reservoir and migrate toward the developing flagella where they assemble into highly ordered layers. It is proposed that scale-scale interactions induce a process of auto-assembly initiating the complex arrangement of scale tiers on the flagellum and cell body.     

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.     

Serial reconstruction of the mitochondrial reticulum in the antarctic flagellate,Pyramimonas gelidicola (Prasinophyceae,Chlorophyta)

Summary A serial reconstruction ofPyramimonas gelidicola McFadden, Moestrup andWetherbee has revealed a large reticulated mitochondrion branching throughout the cell. The possibility of single mitochondria in other members of thePrasinophyceae and the uniformity of the morphology of this organelle within the class is discussed.     

The flagellar apparatus ofMesostigma viride (Prasinophyceae): Multilayered structures in a scaly green flagellate

Mesostigma viride Lauterborn (Prasinophyceae) is the first green flagellate found to have multilayered structures (MLS) in its flagellar apparatus. MLS's were previously known from green algae only in charophycean swarmers, linking theCharophyceae to the origin of land plants, whose male gametes (when flagellated) also possess an MLS.M. viride is, therefore, probably more closely related to the origin of theCharophyceae than any other green flagellate that has been thoroughly studied so far. The occurrence of MLS's in green flagellates and apparently in other algae and protozoans suggests that an MLS occurred in an ancient group of flagellates and has survived in various protistan lines, including the line of green algae related to land plants. The occurrence of a synistosome inM. viride and other of its characteristics suggest that it is more closely related toPyramimonas than to other genera of scaly green flagellates.This work was supported by National Science Foundation Grant DEB-78-03554.     

Flagellar apparatus ultrastructure inMesostigma viride (Prasinophyceae)

The ultrastructure of the flagellar apparatus ofMesostigma viride Lauterborn (Prasinophyceae) has been studied in detail with particular reference to absolute configurations, numbering of basal bodies, basal body triplets and flagellar roots. The two basal bodies are interconnected by three connecting fibers (one distal fiber = synistosome, and two proximal fibers). The flagellar apparatus shows 180° rotational symmetry; four microtubular flagellar roots and two system II fibers are present. The microtubular roots represent a 4-6-4-6-system. The left roots (1s, 2s) consist of 4 microtubules, each with the usual 3 over 1 root tubule pattern. Each right root (1d, 2d) is proximally associated with a small, but typical multi-layered structure (MLS). The latter displays several layers corresponding to the S₁ (the spline microtubules: 5–7), and presumably the S₂—S₄ (the lamellate layers) of the MLS of theCharophyceae. At its proximal origin (near the basal bodies) each right root originates with only two microtubules, the other spline microtubules being added more distally. The structural and positional information obtained in this study strongly suggest that one of the right roots (1d) ofMesostigma is homologous to the MLS-root of theCharophyceae and sperm cells of archegoniate land plants. Thus the typical cruciate flagellar root system of the green algae and the unilateral flagellar root system of theCharophyceae and archegoniates share a common ancestry. Some functional and phylogenetic aspects of MLS-roots are discussed.Dedicated to Prof. DrLothar Geitler on the occasion of his 90th birthday.     

Ultrastructure of the flagellar apparatus in the green flagellateSpermatozopsis similis

The ultrastructure of the flagellar apparatus of the naked, biflagellate green algaSpermatozopsis similis Preisig & Melkonian has been studied in detail using an absolute configuration analysis. The two basal bodies are displaced by 350 nm in the 1/7 o'clock direction and do not overlap proximally. They are interconnected by a principal distal connecting fibre consisting of a bundle of 5–8 nm filaments and possibly two proximal striated connecting fibres. The flagellar root system is cruciate (5-2-5-2 or 4-2-4-2 system) and contains a prominent continuous system I fibre overlying the two opposite two-stranded roots. A system II fibre is absent. Pronounced structural differences have been observed in the flagellar apparatus ultrastructure at two types of flagella orientation: During backward swimming basal bodies are parallel, the distal connecting fibre is extremely contracted; during forward swimming basal bodies assume various angles (from 20° to 180°) and the connecting fibre is about five times longer compared to the contracted state. The function of the connecting fibre as a contractile organelle and the mechanism of its contraction are discussed. On the basis of the flagellar apparatus ultrastructure,Spermatozopsis similis is related toChlamydomonas-type green algae.     

Ultrastructure and freeze-fracture studies of the thylakoids ofMantoniella squamata (Prasinophyceae)

Summary The ultrastructure and the supramolecular organization of the thylakoids of the small green flagellate,Mantoniella squamata, were examined in thin sections and freeze-fracture preparations. The whole chloroplast is tightly packed with thylakoids, which show a pattern of meandering, branching and/or anastomosing membranes. In freeze-fracture preparations only two fracture-faces can be distinguished: the PF- and the EF-face. The PF-face has a much higher particle density than the EF-face (PF: 4086 particles/m²; EF: 865 particles/m²). The EF-face is not as uniform as the PF-face. The areas which are packed with particles probably correspond to closely appressed thylakoid regions or adhesive patches, noticed in thin sections in some areas. The mean particle size on both faces is also different (EF: 10.5 nm; PF: 8.6 nm), but no information about the classification of the particles to special protein complexes is available at this time.Abbreviations chl chlorophyll - EF exoplasmic fracture face - ER endoplasmic reticulum - LHC light-harvesting chlorophyll-protein complex - PF protoplasmic fracture face - PS I photosystem I - PS II photosystem II     

The flagellar apparatus of the scaly green flagellatePyramimonas obovata: Absolute configuration

Summary The flagellar apparatus of the quadriflagellate scaly green algaPyramimonas obovata has been studied in detail and the absolute configuration of the flagellar apparatus has been determined. The flagellar root system is cruciate (4-2-4-2-system). 18 major basal body associated fibrous structures connect the four basal bodies with each other. Each basal body is linked to an adjacent basal body by a unique set of connecting fibres, i.e., the flagellar apparatus does not exhibit 180° rotational symmetry. The flagellar apparatus ofPyramimonas obovata is compared with that of quadriflagellate motile cells of theChlorophyceae sensu Stewart andMattox and the phylogenetic relationships are discussed.     

Ultrastructure of distal flagellar swellings in spermatocytes and spermatids of Ephestia kuehniella Z. (Pyralidae,Lepidoptera)

Summary Development of flagella was investigated by transmission electron microscopy in spermatocytes and spermatids of the Mediterranean mealmoth, Ephestia kuehniella Z. Growing flagella displayed voluminous distal swellings. In short flagella the apical portion of the swellings contained an amorphous, dense accumulation. In more developed flagella a less dense proximal extension of the apical accumulation was formed, which in turn was in contact with the elongating flagellar microtubules. The material of the flagellar tip is interpreted as being a precursor of the axoneme containing mainly tubulin. The material may be converted into the axoneme.     

Two-headed outer- and inner-arm dyneins of Leishmania sp bear conserved IQ-like motifs

Dyneins are high molecular weight microtubule based motor proteins responsible for beating of the flagellum. The flagellum is important for the viability of trypanosomes like Leishmania. However, very little is known about dynein and its role in flagellar motility in such trypanosomatid species. Here, we have identified genes in five species of Leishmania that code for outer-arm dynein (OAD) heavy chains α and β, and inner-arm dynein (IAD) heavy chains 1α and 1β using BLAST and MSA. Our sequence analysis indicates that unlike the three-headed outer-arm dyneins of Chlamydomonas and Tetrahymena, the outer-arm dyneins of the genus Leishmania are two-headed, lacking the γ chain like that of metazoans. N-terminal sequence analysis revealed a conserved IQ-like calmodulin binding motif in the outer-arm α and inner-arm 1α dynein heavy chain in the five species of Leishmania similar to Chlamydomonas reinhardtii outer-arm γ. It was predicted that both motifs were incapable of binding calmodulin. Phosphorylation site prediction revealed conserved serine and threonine residues in outer-arm dynein α and inner-arm 1α as putative phosphorylation sites exclusive to Leishmania but not in Trypanosoma brucei suggesting that regulation of dynein activity might be via phosphorylation of these IQ-like motifs in Leishmania sp.     

Light and electron microscopical studies onHafniomonas gen. nov. (Chlorophyceae,Volvocales), a genus resemblingPyramimonas (Prasinophyceae)

Based on light and electron microscopical studies ofPyramimonas reticulata the genusPyramimonas is shown to contain a number of unrelated flagellates.P. reticulata andP. montana are transferred to the new genusHafniomonas, cells of which differ fromPyramimonas in shape, in the absence of scales and hairs on the body and flagellar surfaces, in details of the chloroplast, the position of the nucleus, the Golgi apparatus, the internal structure of the flagellar apparatus, and in cell division. The prasinophytePyramimonas contains a characteristic association of a large microbody and a rhizoplast, situated on the nuclear surface. A similar association is being found in an increasing number of prasinophycean flagellates, but is absent inHafniomonas, which is considered related to chlorophycean rather than prasinophycean flagellates. The phylogenetic position ofHafniomonas is discussed, based in particular on details of the unique flagellar apparatus.     

The absolute configuration of the flagellar apparatus in zoospores from two species ofLaminariales (Phaeophyceae

Summary We examined the zoospores produced by the unilocular sporangia ofLaminaria digitata (L.) Lamour. andNereocystis luetkeana Post. & Rupr. by serial sectioning to determine the absolute configuration of their flagellar apparatuses. The basal bodies, which are interconnected by three striated bands, lie parallel to the ventral face of the zoospore, and the posterior basal body always is found to the right of the anterior basal body when the cell is viewed from the ventral face, anterior end up. The four rootlets associated with the basal bodies include a major anterior rootlet of about seven microtubules extending from the anterior basal body along the ventral face towards the apex, a five-membered bypassing rootlet that passes ventral to the basal bodies and is connected to the posterior basal body by a posterior fibrous band, and two short rootlets having a single member each, the minor anterior and posterior rootlets. We consider the configuration observed here to be typical of most phaeophycean motile cells. The flagellar apparatus features suggest a considerable phylogenetic difference between thePhaeophyceae and other classes of chlorophyll c-containing organisms.     

Isolation,purification, and characterization of flagellar scales from the green flagellateTetraselmis striata (Prasinophyceae)

Summary Flagellar scales from the green flagellateTetraselmis striata (Prasinophyceae) were isolated, purified by isopycnic cesium chloride-gradient and zonal sucrose gradient centrifugation and their structure and biochemical composition investigated. Three types of flagellar scales were purified to more than 90% purity, a fourth type up to 75% purity. In addition to the previously known types of flagellar scales (pentagonal scales, rod-shaped scales, hair-scales), a novel scale type (i.e., the knotted scales) was discovered. New information about the asymmetric structure of the rod-shaped scales is presented and consequently they are renamed man scales. Flagellar scales consist mainly of carbohydrate (50–70%), significant amounts of protein (11% of dry weight) were found only in pentagonal scales. The main sugars (90%) of the pentagonal and man scales are the unusual 2-keto-sugar acids 3-deoxy-5-O-methyl-2-octulosonic acid (5 OMeKDO), 3-deoxy-2-heptulosaric acid (DHA), and 3-deoxy-2-octulosonic acid (KDO), the knotted scales contain as major sugars galactose and arabinose in addition to KDO and 5 OMeKDO but lack DHA. 13 major polypeptides were identified in flagellar scales by one-dimensional SDS-PAGE, 11 of these are of high molecular mass (>116 kDa). While the majority of polypeptides was found associated with pentagonal scales, at least 4 polypeptides were tentatively assigned to the hair-scales and knotted scales.Abbreviations CSF crude scale fraction - PS pentagonal scales - MS man scales - HS hair-scales - KS knotted scales - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis - DHA 3-deoxylyxo-2-heptulosaric acid - 5 OMeKDO 3-deoxy-5-O-methyl-manno-2-octulosonic acid - KDO 3-deoxy-manno-2-octulosonic acid - GA Golgi apparatus     

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.     

Studies on the ultrastructure and taxonomy of the genusTetraselmis (Prasinophyceae)

Ultrastructural investigations on many isolates ofTetraselmis have revealed that the species have characteristic fine structural features of the pyrenoid and it was proposed (Horiet al., 1982) that the genus be subdivided into four subgenera. In the present study of this series, species of the subgenusPrasinocladia, includingTetraselmis marina, T. verrucosa andT. verrucosa f.rubens, are described in detail.     

Fine structure of the flagellar apparatus of gametesin situ and motile zygotes of the green algaUlothrix flacca var.Roscoffensis (Ulotrichales) (Chlorophyta)

Summary This fine structural study ofUlothrix flacca (Dillw.) ThuretRoscoffensis variety (Berger-Perrot), a marineUlothrix, describes in detail the flagellar apparatus configuration of gametesin situ in the gametangia and in motile zygotes. The gametes's flagellar apparatus shows two basal bodies overlapping at their proximal end at a 30° angle, in an 11/5 o'clock configuration or with a counterclockwise absolute orientation. The basal bodies are interconnected by a non-striated band or capping plate. They are wrapped in their proximal part by an electron-dense sheath and obtured by a bilobed terminal cap. A cruciate microtubular root system having a 4-2-4-2 alternation pattern is present. A striated microtubule associated component (S.M.A.C.) or system I fibres accompany the two membered root R₂. The system II fibres or rhizoplasts along with striated bands associated to the microtubular roots, were not observed and are presumed to be absent.In the motile zygotes, the basal bodies are paired in a cruciate pattern. During the fusion process, two basal bodies, one of each pair, slide in a face to face position with a slight displacement into the 11/5 o'clock direction; the other two make a 30° counterclockwise rotation, thus making a 60° angle between the two basal bodies of each pair instead of 30° in the gamete.After comparison with the flagellar apparatus of other green alga gametes, it is concluded that the taxonomic affinities ofUlothrix flacca var.Roscoffensis, lie with theUlvophyceae sensuStewart andMattox 1978.Abbreviations CP capping plate - ER endoplasmic reticulum - G Golgi body - LG lipid globule - M mitochondria - MS presumed mating structure - N nucleus - R₂,R₄ microtubular roots - SH sheath - SMAC striated microtubule associated component - TC terminal cap - V vacuole - Ve vesicles in the anterior papilla - 1, 2, 1, 2 basal bodies numerotation     

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.