Ultrastructure of Euglena anabaena var. minor (Euglenophyceae)

The freshwater green euglenoid Euglena anabaena var. minor has a pellicle with groove‐ridge articulation, a chloroplast with pyrenoids doubly sheathed by two paramylon caps, and a nucleus with permanently condensed chromosomes and nucleolus. The flagellar apparatus basically resembles that of Euglena. The dorsal root (DR) originates at the dorsal basal body of the emergent flagellum, while both the intermediate root (IR) and ventral root (VR) originate at the ventral basal body of the non‐emergent flagellum. The cytoplasmic pocket is associated with the ventral root/ reinforcing microtubular band. However, ultrastructural characterization of E. anabaena var. minor shows the pocket to consist of five to seven microtubules, and flagellar roots with microtubule configuration of 3–4–6 in the DR‐IR‐VR. The dorsal band microtubules pair at the reservoir‐canal transition level. The doublet microtubules are formed into triplets and doublets at the lower canal level and then make pellicular microtubules at the upper canal level.     

ARE CYTOPLASMIC POCKETS (MTR/POCKET) PRESENT IN ALL PHOTOSYNTHETIC EUGLENOID GENERA?1

In 1985, the existence of a cytoplasmic pocket formed from the reservoir membrane in the photosynthetic euglenoid Colacium was described. A band of reinforcing microtubules (MTR) derived from the ventral flagellar root lined the pocket, and a dense fibrillar mesh was associated with the membrane. A comparison of bodonid cytostomes, colorless euglenoid cytostomes, and the reservoir pocket found in Colacium suggested that the three structures were homologous and that photosynthetic euglenoids arose from phagotrophic ancestors. MTR/pockets have since been reported in other photosynthetic euglenoids, including Euglena, Eutreptia, Eutreptiella, Cryptoglena, Tetreutreptia, and Phacus. We found MTR/pockets in three additional taxa, Lepocinclis, Trachelomonas, and Strombomonas, thereby demonstrating the presence of this complex in representatives of all the major photosynthetic genera. A comparison of the MTR/pocket complex across genera indicated a reduction in structural complexity that was consistent with recent phylogenetic schemes based on molecular characters. Three alternative hypotheses of the origin of MTR/pockets in phototrophic euglenoids are presented and discussed.     

Flagellar systems in the euglenoid flagellates

The flagellar apparatus of euglenoids consists of two functional basal bodies, three unequal microtubular roots subtending the reservoir, and a fourth band of microtubules nucleated from one of the flagellar roots and subtending the reservoir membrane. The flagellar apparatus of some euglenoids may contain additional basal bodies, striated roots ("rhizoplasts"), fibrous roots, striated connecting fibers between basal bodies, layered structures, or various electron-dense connective substances. With the possible exception of Petalomonas cantuscygni, nearly all euglenoids are biflagellate although the length of one flagellum may be highly reduced. The flagellar transition zone and number of basal bodies are highly variable among species. In recent years a cytoplasmic pocket that branches off from the reservoir has been discovered. The microtubules of the ventral flagellar root are continuous with the microtubules which line this pocket. Based on positional and structural similarities, this structure is believed to be homologous with the MTR/cytostome of bodonids. Coupled with other ultrastructural and biochemical data, the fine structure of the flagellar apparatus supports the belief that the euglenoid flagellates are descendant from bodonid ancestors.     

The flagellar root system in the gamete ofAcetabularia mediterranea

Summary Ultrastructural investigation of the flagellar root system ofAcetabularia gametes reveals one type of organization for both male and female gametes. There is a modified cruciate system with four microtubular bands X-2-X-2, with X=4. A prominent distal striated fiber and a small proximal striated fiber connect the flagellar bases. A striated root fiber type I underlies the microtubular root type II, and a short striated root fiber type I underlies the microtubular root type I (terminology ofMelkonian 1980 b). This specific root system has some details in common with theChlamydomonas type, and others with theUlvaphyceae and the siphonalean algaeDerbesia andBryopsis. This might indicate the phylogenetic relationships.     

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.     

Pigments associated with the flagellum ofEuglena gracilis

The pigments associated with the flagellum of the phytoflagellateEuglena gracllis were characterized by HPLC. The pigment pattern of the wild-type strain was compared with a set of white mutants which did not display phototaxis and photoaccumulation in response to blue light. Flagella of the wild type contained FMN and FAD. Two mutants which lacked the stigma but retained a small paraxonemal body (PAB) contained less flavins. The whiteEuglena mutant FB, which retained a residual stigma and also a PAB, and the white phytoflagellateAstasia longa, a close relative ofEuglena, had normal amounts of flagellar flavins. Cells and flagella ofEuglena wild type contained an unldentified pterin-like pigment, called Pt₁₆, which was substantially reduced inAstasia and theEuglena mutants. A third pigment, designated P₅₂₈ with major absorption at 528 nm and fluorescence emission at 550 nm was present mainly in flagella. The association of the three pigment types with flagella and their respective alterations in the white strains indicates their possible role in photoreception. Dedicated to Pill-Soon Song on the occasion of his 60th birthday.     

Cryptobia udonellae sp. n. (Kinetoplastidea: Cryptobiida) - parasites of the excretory system of Udonella murmanica (Udonellida)

A new cryptobiid flagellates, Cryptobia udonellae sp. n., is described from the excretory channels of Udonella murmanica. The body of flagellates is spindle-shaped. The flagellar pocket is subapical. Two flagella emerge from the pocket. One flagellum turns anterior and is forward-directed; the other flagellum is directed posterior and close to the ventral cell surface. The ventral groove is well developed. The cytostome opens just anterior to the flagellar pocket. The cytostome leads to the short cytopharynx. In the excretory channel of worms the flagellates C. udonellae sp. n. are attached to microvilli of epithelium or lay free in the lumen. Both flagellates have been studied with TEM. The unusual parasite system which involves organisms of four different phylums of animals has been described for the first time.     

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.     

The fine structure of motile cells in the generaUlvaria andMonostroma,with special reference to the taxonomic position ofMonostroma oxyspermum (Ulvophyceae,Chlorophyta)

The zoospores and isogametes ofUlvaria obscura var.blyttii, the isogametes ofMonostroma bullosum, and the anisogametes ofM. grevillei have a flagellar apparatus with counterclockwise absolute orientation and terminal caps, and therefore belong to theUlvophyceae. On the basis of the absence or presence of body scales and the morphologies of certain flagellar apparatus components,Ulvaria obscura var.blyttii is retained in theUlvales, whileM. bullosum, M. grevillei andM. oxyspermum are referred to theUlotrichales. Differences in scale morphology, certain flagellar apparatus components, and early thallus ontogeny support the transfer ofM. oxyspermum to the genusGayralia. Mating structures and their positional relationships within the cell are described from the gametes examined. A plasmalemma-associated plaque that may be a degenerate mating structure occurs in someG. oxysperma motile cells.     

The flagellar root system of zoospores of the green algaChlorosarcinopsis (Chlorosarcinales) as compared withChlamydomonas (Volvocales)

The flagellar root system of zoospores in two species ofChlorosarcinopsis (C. minuta andC. spec.) has been studied in detail. The biflagellate zoospores show a cruciate root system, two of the four microtubular roots containing two microtubules, the other two four microtubules. The flagellar apparatus is otherwise identical with that ofChlamydomonas reinhardi as described byRingo (1967). Evidence is presented that the genusChlamydomonas is characterized by a bilateral symmetric root system (4-2-4-2) rather than a system with four equally numbered roots (i.e. 4-4-4-4). It is suggested that a root system with four identical cruciate roots is not present in any biflagellate algal cell. The taxonomic significance of cruciate root systems in green algae is discussed refering to the identical root systems ofChlorosarcinopsis andChlamydomonas.     

ENTOSIPHON SULCATUM (EUGLENOPHYCEAE): FLAGELLAR ROOTS OF THE BASAL BODY COMPLEX AND RESERVOIR REGION1,2

The flagellar root system of Entosiphon sulcatum (Dujardin) Stein (Euglenophyceae) is described and compared with kinetoplastid and other euglenoid systems. An asymmetric pattern of three microtubular roots, one between the two flagellar basal bodies and one on either side (here called the intermediate, dorsal, and ventral roots), is consistent within the euglenoid flagellates studied thus far. The dorsal root is associated with the basal body of the anterior flagellum (F₁) and lies on the left dorsal side of the basal body complex. Originating between the two flagellar basal bodies, and associated with the basal body of the trailing flagellum (F₂), the intermediate root is morphologically distinguished by fibrils interconnecting the individual microtubules to one another and to the overlying reservoir membrane. The intermediate root is often borne on a ridge projecting into the reservoir. The ventral root originates near the F₂ basal body and lies on the right ventral side of the cell. Fibrillar connections link the membrane of F₂ with the reservoir membrane at the reservoir-canal transition level. A large cross-banded fiber joins the two flagellar basal bodies, and a series of smaller striated fibers links the anterior accessory and flagellar basal bodies. Large nonstriated fibers extend from the basal body complex posteriorly into the cytoplasm.     

Zoospore ultrastructure in species ofTrebouxia andPseudotrebouxia (Chlorophyta)

Zoospore ultrastructure (incl. flagellar apparatus) has been investigated in three species ofTrebouxia (T. glomerata, T. erici, T. pyriformis) and one species ofPseudotrebouxia (P. impressa) using an absolute configuration analysis. Zoospores in all taxa studied are nearly identical in ultrastructure and exhibit a very distinctive disposition of cell organelles: cells are naked, biflagellate and considerably flattened along the plane of flagellar beat, the single contractile vacuole is located anteriorly in the ventral region of the cell, the nucleus is anteriorly to centrally located in the dorsal region of the cell. A single dictyosome is located close to the anterior, ventral edge of the nucleus. The chloroplast occupies a posterior position in the cell and usually has an anterior profile in the left region of the cell. There are two branched mitochondria per cell or a single mitochondrial reticulum with profiles anterior to the nucleus (in the dorsal region of the cell), and posterior to the nucleus. In zoospores ofTrebouxia spp. the posterior mitochondrial profile is associated with a microbody, inP. impressa zoospores the anterior mitochondrial profiles are associated with a microbody. The zoospores contain a distinctive system of three ER-cisternae: one system links to both basal bodies and extends to the nucleus, the other two systems subtend the plasmamembrane on the left and right broad cell surfaces and extend to the posterior region of the cell. The flagellar apparatus is structurally identical to that previously described for zoospores ofFriedmannia israelensis and exhibits basal body displacement by one basal body diameter into the 11/5 o'clock direction, a non-striated distal connecting fiber, a cruciate microtubular root system lacking system I fibers and presence of a single system II fiber which connects the basal bodies with the nucleus and runs parallel to one of the ER-strands. The left flagellar roots (X-roots) are subtended by a complex set of amorphous and striated material that connects each left root with both basal bodies.—This study demonstrates the close systematic relationship between the phycobiontsTrebouxia andPseudotrebouxia and the generaFriedmannia, Pleurastrum, andMicrothamnion and supports recent classification schemes which place all these taxa into a single order separate from otherChlorophyta. Dedicated to Prof. DrElisabeth Tschermak-Woess on the occasion of her 70th birthday.     

The Cytostome of Trypanosoma cruzi and T. conorhini*

SYNOPSIS. A cytostome is described in culture forms and in intracellular stages of Trypanosoma cruzi and in culture forms of T. conorhini. The organelle opens into the flagellar pocket or close to it and runs deeply into the cell.     

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.     

The flagellar root system inHeterosigma akashiwo (Raphidophyceae)

Summary The flagellar apparatus of 3 isolates ofHeterosigma akashiwo (Hada) Hada has been studied by serial sectioning. The two basal bodies lie at almost right angles to one another, but in a different plane, and are interconnected by an extensive root system. This consists of three roots (i) a massive cross-banded fibrous root (= rhizoplast) which extends from near the proximal ends of both basal bodies to the anterior surface of the nucleus, (ii) a compound microtubular root with a layered structure, associated with the hairy anterior flagellum and extending to the anterior surface and (iii) the rhizostyle which passes between the two basal bodies leading anteriorly to a vesicle in the flagellar groove region and following the nucleus posteriorly terminating deep in the cytoplasm. Both the characteristic arrangement of the basal bodies and the presence of the complex layered structure are characteristic of theRaphidophyceae. The broad microtubular root, however, to which the layered structure is attached, appears to be characteristic of nearly all heterokont algae, fungi and protozoa so far examined. Thus, our findings have important implications on phylogenetic relationships within the heterokonts and lead us to question whether some of the present classes such as theChrysophyceae andXanthophyceae are indeed natural groups.     

Comparative cytology and taxonomy of theUlvaphyceae II. Ulvalean characteristics of the stephanokont flagellar apparatus ofDerbesia tenuissima

Summary The stephanokont flagellar apparatus of the zoospores ofDerbesia tenuissima (De Not.) Crouan is examined and compared to the flagellar apparatuses of other green algae. The flagella ofDerbesia are attached to two of three bands which lie at the junction of the body and papilla. Serial longitudinal and cross sections reveal that the basal bodies are attached to the bands along their sides and at their proximal ends. The bands are not striated in any plane. The lack of striation in the bands and the partial covering of the proximal end of the basal bodies by one of the bands closely resemble the type of flagellar connection system described as the Bryopsis-type byMelkonian (1980). Zoospores of ulvalean green algae also possess these features, suggesting that green siphons are phylogenetically related to theUlvales. It is proposed that green siphons be tentatively classified in theUlvaphyceae rather than in theChlorophyceae orCharophyceae.This work supported by NSF Grant DEB 78-03554.     

Cytochromec oxidase ofEuglena gracilis: Purification,characterization, and identification of mitochondrially synthesized subunits

Cytochromec oxidase was purified from mitochondria ofEuglena gracilis and separated into 15 different polypeptide subunits by polyacrylamide gel electrophoresis. All 15 subunits copurify through various purification procedures, and the subunit composition of the isolated enzyme is identical to that of the immunoprecipitated one. Therefore, the 15 protein subunits represent integral components of theEuglena oxidase. In anin vitro protein-synthesizing system using isolated mitochondria, polypeptides 1–3 were radioactive labeled in the presence of [³⁵S]methionine. This further identifies these polypeptides with the three largest subunits of cytochromec oxidse encoded by mitochondrial DNA in other eukaryotic organisms. By subtraction, the other 12 subunits can be assigned to nuclear genes. The isolatedEuglena oxidase was highly active withEuglena cytochromec ₅₅₈ and has monophasic kinetics. Using horse cytochromec ₅₅₀ as a substrate, activity of the isolated oxidase was rather low. These findings correlate with the oxidase activity of mitochondrial membranes. Again, reactivity was low with cytochromec ₅₅₀ and 35-fold higher with theEuglena cytochromec ₅₅₈. The data show that the cytochromec oxidase of the protistEuglena is different from other eukaryotic cytochromec oxidases in number and size of subunits, and also with regard to kinetic properties and substrate specificity.Abbreviations kDa kilodalton - PAGE polyacrylamide gel electrophoresis - SDS sodium dodecyl sulfate - TN turnover number     

ENTOSIPHON SULCATUM (EUGLENOPHYCEAE): FLAGELLAR ROOTS OF THE BASAL BODY COMPLEX AND RESERVOIR REGION1,2

The flagellar root system of Entosiphon sulcatum (Dujardin) Stein (Euglenophyceae) is described and compared with kinetoplastid and other euglenoid systems. An asymmetric pattern of three microtubular roots, one between the two flagellar basal bodies and one on either side (here called the intermediate, dorsal, and ventral roots), is consistent within the euglenoid flagellates studied thus far. The dorsal root is associated with the basal body of the anterior flagellum (F₁) and lies on the left dorsal side of the basal body complex. Originating between the two flagellar basal bodies, and associated with the basal body of the trailing flagellum (F₂), the intermediate root is morphologically distinguished by fibrils interconnecting the individual microtubules to one another and to the over lying reservoir membrane. The intermediate root is often borne on a ridge projecting into the reservoir. The ventral root originates near the F₂ basal body and lies on the right ventral side of the cell. Fibrillar connections link the membrane of F₂ with the reservoir membrane at the reservoircanal transition level. A large cross-banded fiber joins the two flagellar basal bodies, and a series of smaller striated fibers links the anterior accessory and flagellar basal bodies. Large nonstriated fibers extend from the basal body complex posteriorly into the cytoplasm.     

THE FLAGELLAR APPARATUS AND RESERVOIR/CANAL CYTOSKELETON OF CRYPTOGLENA PIGRA (EUGLENOPHYCEAE)1

The flagellar apparatus and reservoir cytoskeleton of Cryptoglena pigra Ehrenberg are described. Three flagellar roots are associated with the two basal bodies. The four-membered dorsal root arises from the dorsal basal body and extends anteriorly following the reservoir membrane. At the base of the reservoir the dorsal root nucleates a large microtubular group termed the dorsal band. The dorsal band continues anteriorlhy between the reservoir and eyespot and is continuous with the microtubules of the canal and ultimately the pellicle. The ventral basal body is associated with two roots. The four-membered intermediate root proceeds anteriorly and extends the length of the reservoir. The seven-to eight-membered ventral root projects anteriorly along the reservoir membrane and bends away from the reservoir. At this point, the microtubules of the ventral root line a cytoplasmic pocket and are termed the MTR (reinforcing microtubules). The canal region is composed of longitudinal microtubules surrounded by two semicircles of microtubles. Ultimately, the fifteen ridges of the canal give rise to the pellicular ridges.