SPERMATOGENESIS IN COLEOCHAETE PULVINATA (CHAROPHYCEAE): EARLY BLEPHAROPLAST DEVELOPMENT

Transmission electron microscopy of serial thin sections was used to reconstruct several early developmental stages of the blepharoplast in Coleochaete pulvinata spermatids. These were compared to published studies of blepharoplast development in Charales and the closest relatives of charophycean green algae among embryophytes, i.e., hornworts and liverworts. Bicentriolar centrosomes such as occur in bryophytes and fern allies were not observed in Coleochaete. Centriole replication in C. pulvinata was orthogonal as in Charales. The resulting two daughter centrioles were oriented perpendicularly and joined proximally by electron-dense material. Their orthogonal relationship was maintained throughout blepharoplast development by a massive, banded connective which appeared early. In spermatids of hornworts and liverworts, a multilayered structure (MLS) develops in association with two centrioles destined to become flagellar basal bodies. When the MLS of these lower land plants is sectioned at right angles to the long axis of the microtubular layer, the MLS is observed to lie beneath cross sections of both centrioles. In contrast, when developing MLSs of C. pulvinata and Charales are similarly sectioned, they occur beside a cross section of just one of the two centrioles. In C. pulvinata (as in other charophytes), MLS lamellae are oriented at a 90-degree angle to the long axis of the S₁ microtubules from the beginning. This contrasts with the 40–45 degree angle between the MLS lamellae and S₁ microtubules universally reported for archegoniates. In early C. pulvinata spermatids, spline microtubules are closely associated with an anterior mitochondrion having a low stromal density and few cristae. An anterior mitochondrion is typically associated with blepharoplast development in hornworts and liverworts, but has not previously been reported to occur in Coleochaete or any other charophycean alga. In Coleochaete, as in hornworts and liverworts, but unlike Charales, structure of mature blepharoplasts reflects early blepharoplast ontogeny. Very little change in positional relationships among blepharoplast components (flagella, connective, MLS) occurs during development. These character-state differences are of importance in cladistic analyses of charophycean algae and lower land plants.     

An ultrastructural re-examination of putative multilayered structures inTrentepohlia aurea

Summary Due to recent interest in the significance of multilayered structures (MLSs) and MLS-like structures in green algal and land plant systematics and evolution, the putative MLSs ofTrentepohlia aurea swarmers were re-examined using conventional TEM of serial thin sections, and high-voltage electron microscopy (HVEM) of thick sections. Features not previously reported to occur in trentepohlialean algae include a structure connecting MLSs to adjacent basal bodies, the lamellar nature of the sub-microtubular layer of the MLS, and the presence of appendages on microtubules overlying the lamellar strip. Reasons are given for concluding that the MLSs ofTrentepohlia and other members of theTrentepohliales are possibly homologous to MLSs ofGlaucophyceae and the MLS-like structures ofFriedmannia. It is proposed that increased attention to the details of MLS structure may be crucial to an understanding of the divergence of main lines of green algal evolution.     

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.     

THE OCCURRENCE AND PHYLOGENETIC SIGNIFICANCE OF A MULTILAYERED STRUCTURE IN COLEOCHAETE SPERMATOZOIDS

Spermatozoid-forming cells of Coleochaete scutata were found in packets of four arranged in concentric internal bands. Spermatozoids, which occur singly in antheridial cells, are spherical to ovoid, approximately 7 μm long by about 3.9 μm wide. As compared to relatively unspecialized zoospores, male gametes undergo a number of specialized cellular changes during development. The spherical nuclei and cytoplasm of mature spermatozoids are increased in density. Posterior plastids are reduced and contain large starch grains. Many small mitochondria are clustered near the cell anterior. The plasmalemma is covered with a layer of flattened, diamond-shaped scales, while body scales of zoospores are pyramidal. The two flagella of both zoospores and spermatozoids are covered with flattened, diamond-shaped scales and hairs. The spermatozoids contain an anterior multilayered structure (MLS) structurally similar to, though smaller than, the MLS observed in zoospores. An asymmetrical cytoskeleton consisting of a band of 30–45 microtubules extends from the MLS down one side of the spermatozoid close to the plasmalemma. An immature MLS was observed in an early stage of spermatozoid development. The finding of an MLS and asymmetrical cytoskeleton in specialized male gametes as well as relatively unspecialized zoospores of Coleochaete strengthens assumptions of homology between MLSs of green algal reproductive cells and those found in flagellated spermatozoids of archegoniate plants. The structure of the spermatozoid of Coleochaete supports the hypothesis that this alga may be relatively close to the phylogenetic line which led directly to archegoniates.     

Ultrastructural characterization of the locomotory cytoskeletal system of the spermatozoid inGinkgo biloba

Summary The development of the locomotory cytoskeletal system of sperm is carefully coordinated with the development of the sperm inGinkgo biloba. Here we report further ultrastructural characterization of the locomotory cytoskeletal system in the developing spermatid and mature spermatozoid, particularly with respect to the initiation and early development of the flagellar apparatus. A multilayered structure (MLS) assembles from an electron-dense matrix that self-organizes after blepharoplast breakup and then further elongates. At the tail of the assembling MLS, the spline microtubules connect to an anterior beak of the nuclear envelope. Nuclear-pore complexes are found on the nuclear envelope close to this beak. The mitochondria which elongate and line up one behind the other are tightly associated with the MLS. The MLS ofG. biloba is composed of an upper layer of parallel spline microtubules and a lower layer consisting of a fibrous lamellar strip composed of paralled fibers about 9 nm in diameter. Higher-magnification images show that the fully assembled fibers of the lamellar strip consist of subunits which suggest that protofilaments are involved in the assembly processes. A unique cytoskeletal system of the spermatozoid inG. biloba is given by the anterior bundle of microtubules. This bundle, in which microtubules are arranged parallel to each other, forms between the plasmalemma and the MLS and is about 214–392 nm in cross section. These microtubules expand spirally along the MLS band. Other details of cellular fine structure of the mature spermatozoid are described.     

Studies on woloszynskioid dinoflagellates IX: ultrastructure,cyst formation and phylogeny of the ‘red-snow’ alga Borghiella pascheri (Suchlandt) Moestrup (= Glenodinium pascheri,Woloszynskia pascheri,Gyrodinium nivalis)

Red snow caused by dinoflagellates is a phenomenon rarely reported, described from the European Alps from 1914 onwards, and subsequently observed outside Europe on several occasions in Ontario, Canada. Considerable taxonomic confusion exists regarding the identity of the organism(s) causing red snow, but the most recent occurrence in 2016 in Ontario has now allowed detailed studies, including LM, SEM, TEM and molecular sequencing of the causative species. We conclude that the two species originally described as the cause of red snow, Glenodinium pascheri and Gyrodinium nivale, are synonymous and that the appropriate name for the organism is Borghiella pascheri (syn. Woloszynskia pascheri) as suggested by Moestrup & Calado in the recent volume of the Süsswasserflora. The central part of Borghiella pascheri cells is tomato red and filled with numerous organelles, whose ultrastructure indicates modified chloroplasts. Lack of cultures has prevented chemical characterization of the red pigment. Formation of temporary cysts was common in the samples. Transformation of the motile cells into temporary cysts was followed in detail, and the cysts were shown to be surrounded by the fused inner membranes of the amphiesmal vesicles, which became the cell membrane of the cysts, covered by the fused pellicle precursors. The cell membrane from the motile cell was discarded together with the outer amphiesmal vesicle membrane and the thin thecal plates, and the temporary cysts were therefore not surrounded by any pattern of vesicles. Sexual reproduction resulted in the formation of hypnozygotes. Although the species possessed several unusual features, DNA sequencing showed it to belong to Borghiella. The culture established in 1965 from the Botanical Garden in Göttingen, Germany and generally identified as Woloszynskia pascheri belongs to a separate species of Borghiella, to be described separately.The occurrence of red snow caused by dinoflagellates is discussed.     

Ultrastructural studies of spermatogenesis inAnthocerotophyta V. Nuclear metamorphosis and the posterior mitochondrion ofNotothylas orbicularis andPhaeoceros laevis

Summary Ultrastructural observations reveal that the spermatozoids of the hornwortsNotothylas andPhaeoceros contain two mitochondria and not one as described previously. Mitochondrial ontogeny and nuclear metamorphosis during spermiogenesis in these plants differ from all other archegoniates. The discovery that the posterior region of the coiled nucleus (when viewed from the anterior aspect) lies to the left of the anterior, in striking contrast to the dextral coiling of the nucleus of spermatozoids of other embryophytes, underlines the isolated nature of the hornworts among land plants. As the blepharoplast develops, the numerous ovoid mitochondria initially present in the nascent spermatid fuse to form a single elongated organelle which is positioned subjacent to the MLS and extends down between the nucleus and plastid. At the onset of nuclear metamorphosis, the solitary mitochondrion has separated into a larger anterior mitochondrion (AM) associated with the MLS and a much smaller posterior mitochondrion (PM) adjacent to the plastid. The PM retains its association with the plastid and both organelles migrate around the periphery of the cell as the spline MTs elongate. By contrast, in moss spermatids, where mitochondria undergo similar fusion and division, the AM is approximately the same size as the PM and the latter is never associated with the spline. As in other archegoniates, except mosses, spline elongation precedes nuclear metamorphosis in hornworts. Irregular strands of condensed chromatin compact basipetally to produce an elongated cylindrical nucleus which is narrower in its mid-region. During this process excess nucleoplasm moves rearward. It eventually overarches the inner surface of the plastid and entirely covers the PM.Abbreviations ABB anterior basal body - AM anterior mitochondrion - LS lamellar strip - MLS multilayered structure - MT microtubule - PBB posterior basal body - PM posterior mitochondrion     

Ultrastructure of the spermatozoid of Lycopodium obscurum (Lycopodiaceae)

Ultrastructural observations reveal that the spermatozoid of Lycopodium obscurum is crescent shaped and contains two posteriorly directed flagella that are inserted at the front of the cell. The nucleus is broad and elongated with a narrow posterior projection or nuclear diverticulum. Spline microtubules (MTs) number 180 at their maximum and provide the framework for the cell. These MTs extend from the anterior of the locomotory apparatus and along the outermost surface of the nucleus, with a central shank of 14–17 MTs encircling the cell for at least one-third gyre beyond the nucleus. The two basal bodies are slightly staggered and positioned at the front of the cell over a highly elongated multilayered structure (MLS). The MLS extends laterally around the cell anterior and curves posteriorly over the nucleus. One large anterior mitochondrion is situated subjacent to the MLS, while numerous small mitochondria are scattered near or among the lobes of the single plastid. The plastid rests on the inner nuclear surface and contains numerous large starch grains. This cell differs from that of L. cernuum, the only other species of Lycopodium examined to date, in that it is more elongated and has an anterior-posterior orientation of the nucleus, basal bodies, MLS, and spline. Comparisons with coiled gametes of bryophytes and Selaginella suggest that some degree of coiling and cell streamlining may be ancestral in archegoniate spermatozoids.     

COMPARATIVE ULTRASTRUCTURAL STUDIES OF SPERMATOGENESIS IN THE METZGERIALES (HEPATOPHYTA) II. THE BLEPHAROPLAST OF BLASIA PUSILLA

As in other hepatics, the young spermatid of Blasia pusilla contains a well-developed blepharoplast comprising a four-layered multilayered structure (MLS) and two overlying dimorphic basal bodies. The asymmetrical spline (S₁ or upper stratum of the MLS) numbers 20 or 21 microtubules (MTs) at its anterior tip and reduces to eight at the posterior limit of the lamellar strip (LS). Behind this the shank of the spline is five or six tubules in width over most of its length, approximately one revolution of the circumference of the gamete. The three-microtubule spline aperture underlies the anterior basal body and like those of most hepatics, it is closed at its anterior end. The asymmetrical LS (approx. 2.0 μm in length) is characterized by a right-hand posterior notch which lies below the spline aperture at the region of the cartwheel configuration of the anterior basal body (ABB). The staggered dimorphic basal bodies overlap for approximately one third of their lengths. Both lie parallel to the long axis of the spline. As in other hepatics, the ABB (1.2 μm in length) is subapical and comprises an anterior hub extension with progressive rearward additions of lateral, dorsal and ventral triplets. Over most of its length (2.1 μm) the longer posterior basal body (PBB) consists of a distinct central hub and three ventral triplets. Transition zones of both basal bodies contain stellate configurations into which the two central axonemal MTs frequently extend. The blepharoplast of Blasia shows several features in common with leafy, simple thalloid and complex thalloid liverworts. Compared with the few Metzgeriales observed thus far, the LS is less elongate and the basal bodies less staggered. Dimensions of basal body components and spline dimensions, however, are comparable to those of most leafy and thalloid hepatics. Striking similarities with the complex thalloid liverworts include a posterior notch in the LS and a spline aperture three MTs wide.     

ULTRASTRUCTURE OF CEPHALEUROS VIRESCENS (CHROOLEPIDACEAE; CHLOROPHYTA). IV. ABSOLUTE CONFIGURATION ANALYSIS OF THE CRUCIATE FLAGELLAR APPARATUS AND MULTILAYERED STRUCTURES IN THE PRE- AND POST-RELEASE GAMETES

Transmission electron microscopy of pre-release and post-release biflagellate gametes of Cephaleuros virescens has produced comparative data on these cells and on the detailed absolute arrangement of the flagellar apparatus. In all major respects including the presence of two multilayered structures (MLS's) the closely compacted, non-motile but mature pre-release gametes are similar to the mature, free swimming post-release gametes. The elongated shape of the free-swimming gametes differs from the more compact form of the pre-release gametes, but does not reflect a major difference in the arrangement of internal components. The flagella are bilaterally keeled and each keel contains a cylindrical element. Each flagellar base is encircled by a densely staining collar of modified plasmalemma at the point of entry into the apical papilla. The equal anterior flagella enter the papilla from opposite sides; their basal bodies are parallel and overlapping. Each terminates in a densely staining terminal cap. No capping plate is present. Each basal body is associated both with a three-layered MLS, the anterior layer of which becomes a lateral microtubular spline of 2 to 8 microtubules, and with an additional medial compound root of two layers of microtubules (2 over 4 or 5). Both the compound microtubule root and the spline may acquire additional microtubules as they extend distally in close proximity to mitochondria and the plasmalemma. No striated roots, or rhizoplasts, have been observed. Two densely staining plaques are associated with the plasma membrane at specific anterior sites and may be comparable to the presumptive mating structures seen in other green algal motile cells. The reversed bilateral symmetry of the cells produces two possible arrangements of the flagellar apparatus, namely, a 11/5 (or left-handed) arrangement or a 1/7 (or right-handed) arrangement. Only 11/5 cells have been found. Despite the presence of distinct multilayered structures, some aspects of the gametes of Cephaleuros quite closely resemble the cruciate motile cells of algae now regarded by some authors as typical of Ulvophyceae, sensu Stewart and Mattox.     

ULTRASTRUCTURE OF CEPHALEUROS VIRESCENS (CHROOLEPIDACEAE; CHLOROPHYTA). III. ZOOSPORES

Transmission electron microscopic examination of Cephaleuros virescens Kunze growing on leaves of Camellia spp. and Magnolia grandiflora L. indicates that unreleased zoospores in mature zoosporangia are similar to those produced by the related genus Phycopeltis epiphyton Millardet and unlike the quadriflagellate motile cells produced by taxa in other families of Chlorophyta. The zoospores bear four smooth isokont bilaterally “keeled” flagella containing typical “9 + 2” axonemes and lacking scales. Flagellar insertion is apical and the parallel basal bodies overlap laterally at two levels. A cross section through the four basal bodies shows a trapezoidal arrangement wherein the two upper (anterior) basal bodies are closer together than are the lower (posterior) two. Serial sections indicate that diagonally opposing upper and lower basal bodies anchor flagella which emerge from the same side of the apical papilla. Each of the four basal bodies is associated with a microtubular spline which extends beneath the plasmalemma to the posterior end of the zoospore. A distinct multilayered structure is associated with each of the lower basal bodies. A nucleus, mitochondria (two of which are closely associated with the nucleus and spline microtubules), a chloroplast, and cytoplasmic haematochrome droplets are present in each zoospore. Pyrenoids and eyespots are absent. Flagellar insertion is characterized by “reversed bilateral symmetry”; and zoospores with both right-handed and left-handed arrangements are produced. The ultrastructure of the zoospores clearly indicates that: 1) the mode of flagellar insertion: 2) morphology, number, and arrangement of multilayered structures, and 3) bilaterally keeled flagella are characteristic of the Chroolepidaceae.     

Observations on the morphology,reproduction and cytology of stigeoclonium pascheri (vischer) cox et bold (chaetophorales-chlorophyceae) and their bearing on the validity of the genus caespitella vischer

The present communication deals with the morphology, reproduction and cytology of Stigeoclonium pascheri (formerly described as Caespitella pascheri Vischer). Effects of various concentrations of nitrate and variations in intensity and duration of light period in 24 hr LD cycles were studied on cultures of this alga in relation to the type and extent of branching and occurrence or absence of hair formation. It is concluded that these traits are not of taxonomic significance and are influenced largely by environmental conditions. The present study shows a large overlapping of morphological characters between the genera Stigeoclonium and Caespitella and supports Cox & Bold's abandonment of the latter generic name and its merger into the genus Stigeoclonium. The chromosome number determined for this alga is n = 6 which indicates that the Indian strain of this alga is a distinct cytotype from the one originally isolated by Vischer.     

COMPARATIVE ULTRASTRUCTURAL STUDIES OF SPERMATOGENESIS IN THE METZGERIALES (HEPATICAE). I. THE BLEPHAROPLAST OF PALLAVICINIA LYELLII

The blepharoplast in the young spermatid of Pallavicinia is similar to that of other hepatics in that it comprises a four-layered multilayered structure (MLS) and two staggered, dimorphic basal bodies. The spline, approximately 40 μm in length and extending through nearly two full gyres, comprises 20 parallel microtubules at its anterior end and narrows to 17 at the posterior limit of the subjacent lamellar strip (LS). Behind this, the spline shank, approximately 32 μm in length, is reduced to six tubules. The LS curves around the spermatid, following the anterior one-third of the first gyre of the spine, and is approximately 7.5 μm in length, the longest yet recorded for the bryophytes. It is spatulate in outline, equaling the width of the spline anteriorly but tapering steeply from the right-hand side behind the anterior basal body (ABB). It then extends posteriorly as a narrow strip beneath the left-hand margin of the spline. The basal bodies of the greatly staggered flagella are nonoverlapping and separated by a distance of about 4.4 μm. The subapical ABB and PBB measure (including the ventral triplet extensions and transition zones) 1.2 μm and 2.4 μm in length, respectively. A short, narrow aperture equaling one tubule-diameter in width is located in the spline directly beneath the ABB. The anterior mitochondrion is about 7 μm long and follows the outline of the overlying LS, while a cupshaped posterior mitochondrion is appressed to the plastid. Comparisons with other taxa indicate that major distinguishing features of metzgerialian blepharoplasts are highly staggered, nonoverlapping basal bodies, greatly elongate anterior mitochondria, and six-tubule shanks. Great differences between the spermatids suggest wide phylogenetic discontinuities between the genera of the Metzgeriales.     

THE ULTRASTRUCTURE OF MULTILAYERED STRUCTURES ASSOCIATED WITH FLAGELLAR BASES IN MOTILE CELLS OF TRENTEPOHLIA AUREA1

An ultrastructural study of motile cell development in the green alga Trentepohlia aurea has revealed the presence of multilayered structures (MLS) associated With flagellar bases. These MLS are ultrastructurally similar to MLS described in pteridophyte and bryophyte sperm and in the zoospore of the green algae Coleochaete and Klebsormidium. However, 2 MLS are found in each biflagellate motile cell of T. aurea, while other previously described MLS occur singly in biflagellate motile cells. In addition, the MLS of T. aurea consist of fewer microtubules and are structurally simpler than most other MLS described. The MLS of Trentepohlia may represent a stage in the evolutionary development of the MLS of land plants. The presence or absence of the MLS in motile cells of green algae may be a useful character in phylogenetic studies.     

FEEDING APPARATUS OF THE COLORLESS FLAGELLATE KATABLEPHARIS (CRYPTOPHYCEAE)1

The feeding apparatus of Kalablepharis ovalis (isolated from a freshwater impoundment in Colorado) and Katablepharis clone G-2 (isolated from the littoral of the Black Sea near Yalta in the Crimea) consists of inner and outer oval-shaped arrays of microtubules that begin at the anterior end of the cell and pass into the posterior of the cell. Each array of microtubules contains groups of microtubules with two to eight microtubules per group depending on the position of the array in the cell. A specialized area of the plasma membrane, the mouth, occurs at the anterior end of the cell. The mouth is oval with the long axis oriented dorsoventrally and consists of a raised ridge surrounding a central depression. The anterior end of the microtubules of the inner and outer arrays supports the raised ridge of the mouth. In freeze-fracture replicas, the protoplasmic face of the plasma membrane contains intramembrane particles on the raised ridge of the mouth. Three small membrane-cisternae occur on the protoplasmic side of the plasma membrane in the area of the mouth. Katablepharis clone G-2 also has five or six additional large membrane-cisternae associated with the inner microtubular array in the anterior portion of the cell. These larger membrane-cisternae do not occur in K. ovalis. Vesicles with electron-dense contents occur in association with the microtubular arrays. Katablepharis ovalis has a second type of vesicle containing a single-membrane profile associated with the microtubule arrays. The structure of the microtubular arrays in Katablepharis is compared with similar structures in suctorian ciliates and dinoflagellates.     

SPERMATOGENESIS IN LYCOPODIUM: THE MATURE SPERMATOZOID

The mature spermatozoid of Lycopodium cernuum is a blunt ended, fusiform cell, 8–10 μm long by 4–5 μm wide. A multilayered structure (MLS) and a subtending anterior mitochondrion are located at the anterior of the cell. The MLS is coiled through 1–1.5 gyres in a shallow sinistral helix around the periphery of the cell. The MLS would be triangular in outline if unwound and laid flat, about 1.4 μm wide, 7.5–8 μm long, and 80 nm thick. The MLS comprises four layers, S₁–S₄. The S₁ forms the spline, a supportive sheet of microtubules; the S₂, lamellate in younger stages, is an homogeneous, darkly staining layer in the mature sperm; the S₃ and S₄ retain their lamellate appearance and are delimited by lateral connections. Approximately 200 S₁ microtubules extend posteriorly from the MLS at about 45° to the MLS long axis and form a partial sheath around the nucleus. The two basal bodies are located on opposite sides of the cell external to the MLS. Each is tangential to the curve of the MLS and surrounded by a globular matrix. At their attachment, the axonemes are oriented laterally and are antiparallel to each other. Distally, the flagella, each about 38 μm long, trail behind the cell as it swims. The nucleus is roughly ovoid, about 4 μm diam, and centrally or sometimes laterally located. The greater volume of the nucleus is occupied by condensed, amorphic chromatin. Cavities within the chromatin are often seen to contain spheroidal inclusions that have two differently staining regions. The inclusions are also located at the periphery of the chromatin. The posterior of the cell is occupied by several small mitochondria and an amyloplast, about 2 μm diam containing numerous starch grains.     

Changes in the absolute configuration of the basal/flagellar apparatus and evidence of centrin during male gametogenesis in Chara contraria var. nitelloides (Charales, Charophyta)

The absolute configurations of the basal/flagellar apparatus during male gametogenesis of Chara contraria var. nitelloides (Charales, Charophyta) were carefully analysed. Emphasis was placed on the changes in the angles and lengths of the basal bodies, the microtubular root angles and the development of the distal as well the proximal connecting fibers. Six principal stages were recognized: a) parallel, non-axonemal, developing basal bodies connected by a non-striated, proximal fiber; b) non-parallel, non-axonemal, mature basal bodies connected by a developing, striated, distal fiber; c) non-parallel, axonemal basal bodies connected by a fully developed, striated, distal fiber; d) opposite, axonemal basal bodies not connected by fibers, e) axonemal basal bodies not connected by fibers and directed backwards and f) parallel, axonemal basal bodies not connected by fibers. A headpiece, a 3-membered root and a reduced multilayered structure developed during ontogeny. The initial parallel disposition of the basal bodies, the initial lack of MLS and the presence of only two microtubular roots from the very inception of the basal apparatus development, suggest a Mamiella-like ancestor for Charales. Ontogenetic evidence supports previous ideas in the sense that similarities of sperm morphology of charalean and bryophytan gametes are likely due to convergent evolution. In addition, the present study clearly reveals the presence of centrin in Charales.     

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.     

ULTRASTRUCTURE OF CEPHALEUROS VIRESCENS (CHROOLEPIDACEAE; CHLOROPHYTA). II. GAMETES

Transmission electron microscopic examination of Cephaleuros virescens Kunze growing on leaves of Camellia sp. indicates that gametes are similar to those of Trentepohlia aurea. The gametes bear two, smooth isokont “keeled” flagella containing typical “9 + 2” axonemes and lacking scales. Flagellar insertion is apical and the parallel basal bodies overlap laterally. Each basal body is associated with a separate multilayered structure and component microtubular spline. The latter extends posteriorly beneath the plasmalemma. A nucleus, mitochondria, chloroplasts, and cytoplasmic haematochrome droplets are present. Pyrenoids and eyespots are absent. The subcellular components of C. virescens gametes are comparable to those found in gametes of T. aurea; however, the arrangement of basal bodies and multilayered structures differs slightly from that in T. aurea. Comparison of the fine structure of gametes from Cephaleuros, Phycopeltis, and Trentepohlia clearly indicates that the (1) mode of flagellar insertion, (2) morphology, number, and arrangement of multilayered structures, and (3) keeled flagella are common to these three genera and, thus far, unique among the green algae. Although flagellar insertion is apical, it is not bilaterally symmetrical (sensu stricto), nor is it asymmetrical (cf. Chara and Nitella sperms). The arrangement may be termed “reversed bilateral symmetry” and standardization of the terminology is recommended.