THE ORGANIZATION AND EVOLUTION OF MICROTUBULAR ORGANELLES IN CILIATED PROTOZOA

(1) Ciliated protozoa are viewed as unicellular organisms structured in a hierarchy of organizational levels that include the macromolecular, suborganellar, unit organellar, organellar complex, and organellar system. (2) The ciliate cortex is divided into two major functional regions, the somatic region and the oral region. The fundamental component of the cortex is an organellar complex, the kinetid, whose organizing centre is the kinetosome with which are associated three fibrillar associates diagnostic of ciliates. These three fibrillar associates are the periodically striated kinetodesmal fibril and two microtubular ribbons, the transverse and postciliary ribbons. (3) Somatic and oral kinetids are found to be of three major types: monokinetids are composed of one kinetosome and its fibrillar associates; dikinetids are composed of two kinetosomes and their fibrillar associates; polykinetids are composed of more than two kinetosomes and their fibrillar associptes. (4) The mechanisms underlying kinetid function and development remain largely unexplored. Research into the molecular biology and ultrastructure, especially of mutant forms, should provide basic insights in the near future. (5) The conservation of kinetid structure across major phyla of organisms suggests that this subcellular structure should be useful in phylogenetic analysis despite the concepts of ‘chemical identity’ and ‘organic design’. (6) The evolutionary rate of change of oral features is greater than that of somatic features, probably due to developmental and ecological factors. Nevertheless both cortical regions are constrained by the phenomenon of structural conservatism; that is, the conservation of structure through time is inversely related to the level of biological organization. (7) Eight major groupings of ciliate species are recognized, based on ultrastruc-tural features of the cortex. Several examples of differences between these eight groups and the groups presently recognized are discussed.     

Peculiarities of the brain organization and fine structure in small insects related to miniaturization. 1. The smallest Coleoptera (Ptiliidae)

This paper describes for the first time the organization and fine structure of the brain in the smallest free-living insects Acrotrichis grandicollis, Micado sp., and Nanosella sp. (Ptiliidae, Coleoptera), which were studied using serial histological sections as well as TEM and computer-assisted 3D reconstructions. Some specific structural features related to miniaturization were revealed; the relative size of the brain regions and localization of its structures were analyzed. In spite of the extremely small body size, the brain retains the structure and fine structure typical of larger representatives of related groups, illustrating high conservatism of the brain morphology. Data on the number and size of neurons in the brain of Ptiliidae were obtained. The results obtained confirm and supplement the hypothesis about the factors limiting miniaturization of insects. [The next papers will describe the brain organization in Mymaridae and Trichogrammatidae (Hymenoptera), Corylophidae (Coleoptera), Thripidae (Thysanoptera), and Liposcelidae (Psocoptera).]     

Discotricha papillifera: Structure and Morphogenesis of a Marine Interstitial Ciliate*

SYNOPSIS. Discotricha papillifera Tuffrau, a marine interstitial ciliate, is redescribed with the aid of light, scanning, and transmission electron microscopy from cells collected at a New Hampshire beach. Presence of primitive membranelles as well an an advanced stomatogenesis is demonstrated. The ultrastructure, including a unique membrane-bounded septate structure, is described. The cell is tentatively placed in the nassulid suborder Microthoracina, but affinities with other groups are discussed.     

Cyc17, a meiosis-specific cyclin,is essential for anaphase initiation and chromosome segregation in Tetrahymena thermophila

Although the role of cyclins in controlling nuclear division is well established, their function in ciliate meiosis remains unknown. In ciliates, the cyclin family has undergone massive expansion which suggests that diverse cell cycle systems exist, and this warrants further investigation. A screen for cyclins in the model ciliate Tetrahymena thermophila showed that there are 34 cyclins in this organism. Only 1 cyclin, Cyc17, contains the complete cyclin core and is specifically expressed during meiosis. Deletion of CYC17 led to meiotic arrest at the diakinesis-like metaphase I stage. Expression of genes involved in DNA metabolism and chromosome organization (chromatin remodeling and basic chromosomal structure) was repressed in cyc17 knockout matings. Further investigation suggested that Cyc17 is involved in regulating spindle pole attachment, and is thus essential for chromosome segregation at meiosis. These findings suggest a simple model in which chromosome segregation is influenced by Cyc17.     

The Effect of Starvation on Plastid Number and Photosynthetic Performance in the Kleptoplastidic Dinoflagellate Amylax triacantha

The dinoflagellate Amylax triacantha is known to retain plastids of cryptophyte origin by engulfing the mixotrophic ciliate Mesodinium rubrum, itself a consumer of cryptophytes. However, there is no information on the fate of the prey's organelles and the photosynthetic performance of the newly retained plastids in A. triacantha. In this study, we conducted a starvation experiment to observe the intracellular organization of the prey's organelles and temporal changes in the photosynthetic efficiency of acquired plastids in A. triacantha. The ultrastructural observations revealed that while the chloroplast‐mitochondria complexes and nucleus of cryptophyte were retained by A. triacantha, other ciliate organelles were digested in food vacuoles. Acquired plastids were retained in A. triacantha for about 1 mo and showed photosynthetic activities for about 18 d when measured by a pulse‐amplitude modulation fluorometer.     

Chromatin structure in somatic nucleus of ciliate <Emphasis Type="Italic">Didinium nasutum</Emphasis>

The structural organization of macronuclear chromatin of the ciliate Didinium nasutum was studied. The macronuclear genome of D. nasutum is represented by DNA molecules of subchromosomal size. At interphase, macronuclear chromatin is organized into chromatin of 100–200-nm clumps. Some of these clumps form short, thick fibers that consist of several chromatin clumps. Using the differential staining of nucleic acids on ultrathin sections, we revealed perichromatin fibers and granules on the surface of many chromatin clumps. A 3D model of the spatial distribution of chromatin clumps in the macronucleus was built based on serial ultrathin sections and peculiar features of chromatin spatial organization were studied.     

Regulation of cyanobacterial pigment-protein composition and organization by environmental factors

The coordinate expression of stress-specific genes is a common response of all organisms to altered environmental conditions. In cyanobacteria, the physiological consequences of stress are often reflected in both the ultrastructure of the cell and in photosynthesis-related properties. This review will focus on the alterations in cyanobacterial pigment-protein organization which occur under different growth conditions, and how several molecular genetic aproaches are being used in this laboratory to investigate the regulatory mechanisms underlying these alterations. We will discuss in detail the response to iron starvation, and present a testable hypothesis for the mechanism of thylakoid reorganization mediated by this response.Abbreviations ConA concanavalin A - CP chlorophyll-protein complex - LHCP light harvesting chlorophyll protein - MSP manganese-stabilizing protein     

Feeding response of a benthic copepod to ciliate prey type,prey concentration and habitat complexity

1. Protozoans are important consumers within microbial food webs and, in turn, they represent potential prey for small metazoans. However, feeding interactions within these food webs are rarely characterised and this is especially true for freshwater sediments. 2. We aimed to quantify the feeding links between a freshwater meiofaunal copepod and ciliates in two laboratory experiments. The first experiment addressed the response of Eucyclops serrulatus towards ciliate density and type (two ciliate species of the same genus differing in terms of body size). A second experiment assessed the effect of habitat structure on feeding rates by introducing different structural complexity into the feeding arena. In contrast to the first experiment, which was run only for one time period, this experiment also tested three different total feeding times (4, 7 and 9 h). 3. Eucyclops serrulatus exhibited high ingestion rates, with 3–69 ciliates copepod^?1 h^?1 consumed depending on food concentration, food type and habitat complexity. Copepods exhibited a preference for the smaller ciliate when total ciliate concentration was low, but selected both ciliates equally when food concentrations were medium or high. However, at very high food concentration, Eucyclops preferred the larger ciliate (which was 1/3 of its own body size), suggesting that the longer handling times of the larger prey are rewarding when the large prey is present in high numbers. In terms of total numbers consumed, copepods fed on more small ciliates, but in terms of carbon units both ciliates were selected equally when total prey concentration was low or medium. However, copepods derived more carbon from the larger prey at high and very high prey concentrations (up to 0.7 μgC out of a maximum of 1.1 μgC copepod^?1 h^?1). Habitat complexity influenced the feeding of copepods when it was observed over time. 4. The copepod–ciliate link is well known from the pelagic zone of both marine and freshwater habitats. We have shown its potential importance within the benthos, where it can be influenced by food identity, food quantity and possibly by habitat complexity.     

A new null hypothesis for measuring the degree of plant community organization

An attempt is made to derive a measure for the degree of plant community organization, which would not be based on species co-occurrence and co-abundance. The use of the independent random distribution hypothesis (IRDH) is suggested for this purpose. The hypothesis is expected to be valid, if no deterministic phytosociological-structure-generating mechanism is present. If structural variability is used as a statistic for testing the hypothesis, deviances from the conditions of IRDH (species distributions are independent from each other, environmental gradients are lacking) will be attributable either to species interactions (smaller structural variability than expected), or to environmental heterogeneity (greater structural variability than expected). Structural variability is evaluated as the variance of species diversity, the indexN=exp(H') is used for measuring diversity. The precise measure of the degree of community organizationW is computed as the shift between two empirical distributions:D ^* (VN) or Bootstrap distribution of variance of diversity in the community, andD ^o (VN) or the random community variability distribution, which is evaluated after simulating the IRDH conditions.A satisfactory interpretation can be given to the results of evaluatingW for 11 data sets of 10 relevés each.Abbreviation IRDH Independent random distribution hypothesis     

Spore wall ultrastructure of Polypodiaceae from north-western Argentina

The spore wall ultrastructure of Campyloneurum, Microgramma, Pecluma, Phlebodium, Pleopeltis and Serpocaulum (Polypodiaceae) from north-western Argentina has been studied using transmission electron microscopy (TEM). The exospore is 0.4–3 μm thick, two-layered and variously ornamented in all taxa. The exospore surface is distinctive, but in general ultrastructure the exospore is similar in all species studied. The structural elements of the exospore consist of cavities in the inner part as well as channels with a radial orientation and channels at both sides of the laesura. Variation in the exospore surface was observed in spores at different stages of maturation. The perispore is darkly contrasted and 0.04–2 μm thick. Three different structure types were recognised, including fibrillar, multilamellar and lacunose. Scattered globules and spherules were always present on the perispore surface. The structural variability of the perispore was surveyed within complete sporangia. We concluded that the observed variability may be related to the stage in spore maturation and, consequently, to the stages in perispore differentiation. As the exospore ultrastructure is similar and interpreted as related to functional activity in the studied material, it cannot be used for systematic delimitations at this generic or specific level.     

Ultrastructural Study of an Endosymbiotic Alga and Its Host Ciliate Stentor niger

Stentor niger collected in the suburbs of Hiroshima contained in its cytoplasm several hundreds of endosymbiotic algae and innumerable brownish pigment granules. The body of the ciliate was dark due to a mixture of the green endosymbiotic algae and brown pigment granules. The algae belonged to the genus Chlorella; each was enclosed in a perialgal vacuole and dispersed uniformly in the host cytoplasm from the myoneme layer inward to the center of the ciliate. The cell wall and plasma membrane of the alga enclosed a nucleus, chloroplast, mitochondrion, Golgi complex, accumulation bodies, myelinated vesicles, and many ribosomes. The chloroplast occupied more than half of the volume of the alga and contained a conspicuous pyrenoid. Algal multiplication occurred by two successive divisions of an alga, leading to four autospores within a perialgal vacuole; the walls of the vacuole invaginated to separate the autospores each into its own vacuole. Three types of pigment granules were scattered uniformly throughout the cytoplasm of the ciliate. The ultrastructure of the membranellar region, somatic cortex, and macro- and micronucleus of the ciliate are also described.     

Encystment in the Hypotrichous Ciliate Paraurostyla weissei: Ultrastructure and Cytochemistry1

The cyst wall of Paraurostyla weissei consists of four morphologically distinct layers. It shows an ultrastructure and composition similar to that of the previously described kinetosome-resorbing cysts, and its cytoplasm displays characteristics of “urostylid-type” cysts. Therefore it is possible to consider it a “transition ciliate” between Stichotrichina and Sporadotrichina.     

Polymorphism and Ultrastructure of a Colpodean Ciliate of the Genus Platyophryides Foissner, 1987

ABSTRACT. A ciliate isolated from a pond in Brazil, transformed to a giant form when its food was shifted from a bacterial prey to a ciliate prey. This polymorphism is immediately reversible when the prey ciliates, either Tetrahymena or Colpidium , disappear from the culture medium. By its life cycle, morphology, and ultrastructure, this ciliate belongs to the Class Colpodea. it could belong to the genus Platyophryides Foissner, 1987, except that its micronucleus is not enveloped by the macronuclear membrane. The systematic position of the genus Platyophryides , the validity of the three species in this genus, and the characteristics of the Cyrtolophosidida are discussed.     

Peptidomics: a logical sequel to proteomics

Rapid progress of separation techniques as well as methods of structural analysis provided conditions in the past decade for total screening of complex biologic mixtures for any given class of biomolecules. The present review updates the reader with the modern state of peptidomics, a chapter of chemical biology that deals with structure and biologic properties of sets of peptides present in biologic tissues, cells or fluids. Scope and limitations of currently employed experimental techniques are considered and the main results are outlined. Considerable attention will be afforded to the biologic role of peptides formed in vivo by proteolysis of nonspecialized precursor proteins with other well-defined functions. In conclusion, the connection is discussed between peptidomics and the much more mature and still closely related field of proteomics.     

Ultrastructure of prothoracic glands during larval-pupal development of the tobacco hornworm,Manduca sexta: A reappraisal

The structure of Manduca sexta prothoracic glands was investigated using a protocol that preserves membranes. During the last larval stadium, prothoracic gland cells increase in diameter, volume, protein content, and perhaps number, enhancing their capacity to produce ecdysteroids. The glands' strand-of-cells morphology, their in situ location, the presence of gap junctions between cells, and junctional foot-like structures within cells support previous findings that prothoracicotropic hormone stimulates ecdysteroidogenesis via Ca²⁺-induced Ca²⁺ release. A different method of tissue fixation from that previously used to investigate the ultrastructure of Manduca sexta prothoracic glands has revealed a significantly different ultrastructure. These new findings begin to define roles for endoplasmic reticulum and mitochondria in ecdysteroid synthesis and support the hypothesis that the glands secrete the steroid hormone via exocytosis. The structural dynamics of the glands are discussed in the context of the glands' function during Manduca sexta larvalpupal development. © 1993 Wiley-Liss, Inc.     

Diversity and geographic distribution of ciliates (Protista: Ciliophora)

About 4,500 free-living ciliate morphospecies have been described, applying an average synonymy rate of 20%. We estimate that 83–89% of the ciliate diversity is still undescribed, using the following probabilities: detailed habitat studies suggest that the described number of morphospecies must be doubled: 4,500 → 9,000; this figure has to be increased by about 50% due to species with similar interphase morphology but different resting cysts: 9,000 → 13,500; the genetic and molecular data suggest that this value must be doubled or trebled: 13,500 → 27,000 to 40,000 free-living, biological ciliate species. The knowledge on geographic distribution of ciliates heavily depends on flagship species and statistical analyses because reliable faunistic studies are rare and molecular data are still in its infancy. We present a list of 52 ciliate flagship species as a testable hypothesis, i.e., the hypothesis of restricted distribution of certain ciliate species must be refused when a considerable number of them is found in all or most biogeographic regions. Flagship species and statistical analyses consistently show Gondwanan and Laurasian ciliate communities, suggesting that the split of Pangaea deeply influenced ciliate distribution and rare species play a key role in geographic differentiation. However, there is also substantial evidence for continental, regional, and local endemism of free-living ciliates. The molecular studies usually show a high level of genetic diversity underlying ciliate morphospecies, suggesting that morphologic and molecular evolution may be decoupled in many ciliate species. Molecular studies on ciliate biogeography are at variance, possibly because most are still focusing on single molecular markers. In sum, the data indicate that ciliate biogeography is similar to that of plants and animals, but with an increased proportion of cosmopolites, favouring the moderate endemicity model. Special Issue: Protist diversity and geographic distribution. Guest editor: W. Foissner.     

Simultaneous tests of the preference-performance and phylogenetic conservatism hypotheses: is either theory useful?

The preference-performance and phylogenetic conservatism hypotheses have been postulated to explain the mechanisms driving host-use patterns of phytophagous insects. The preference-performance hypothesis predicts that insects will use plants that provide higher offspring fitness, while the phylogenetic conservatism hypothesis predicts that insects will use phylogenetically closely related plants over more distantly related plants. Although some studies have supported these two hypotheses, others have not. Simultaneous tests of the two hypotheses on more than one species are lacking, and this limits comparative interpretation of previous studies. We undertook a comparative investigation to determine whether preference-performance and/or the phylogenetic conservatism hypothesis can explain host-use patterns of two phytophagous insects, the fruit flies Bactrocera cucumis and B. tryoni. Within a nested, plant phylogenetic framework, oviposition preference and offspring performance of the two fruit fly species were tested on fruits of plant species from across different plant families, from within a family and across cultivars within a species. The results show that both the preference-performance and the phylogenetic conservatism hypotheses can, depending on the host plant taxonomic level, explain host usage patterns in B. cucumis, while neither theory explained the host patterns seen in B. tryoni. In the light of increasing recognition of the complexity of host plant–herbivore relationships, and of ongoing studies which as often as not fail to find support for these theories as those that do, we discuss the limited value of either theory as a basis for future research.     

Electron microscopic studies on the macronuclear development in the ciliate Chilodonella uncinata.

The development of macronuclear anlage in the ciliate Chilodonella uncinata has been studied through electron microscopy. The ultrastructure of macro- and micronuclei is described for comparison. During the first stage of development, when the DNA content of the macronuclear anage increases from 2 C to 32 C, chromosomes are visible as distinct osmiophilic bodies inside the anlage. At the end of the initial polyploidization phase, the chromosomes despiralize, giving rise to long filamentous structures 25 to 50 nm in diameter. The latter show a singular banding pattern, with dense bands 12 to 25 nm thick alternating regularly with less dense interbands. Such an organization has not yet been observed in any other type of nucleus. These filamentous structures have been interpreted as highly despiralized oligotenic chromosomes. During the final stage of macronuclear development, these structures condense into thin chromatin strands and small dense granules; the number of granules increases progressively as the chromatin strands disappear. These small granules very likely fuse amongst themselves to form the chromatin granules of the vegetative macronucleus. No evidence has yet been found for a fragmentation of chromatin in this ciliate, but this problem needs further study. The old macronucleus maintains a normal ultrastructure until a late stage of development of the macronuclear anlage, becoming pycnotic only towards the very end of the latter process.     

Ribosomal proteins: Structure,function, and evolution

The question concerning reasons for the variety of ribosomal proteins that arose for more than 40 years ago is still open. Ribosomes of modern organisms contain 50–80 individual proteins. Some are characteristic for all domains of life (universal ribosomal proteins), whereas others are specific for bacteria, archaea, or eucaryotes. Extensive information about ribosomal proteins has been obtained since that time. However, the role of the majority of ribosomal proteins in the formation and functioning of the ribosome is still not so clear. Based on recent data of experiments and bioinformatics, this review presents a comprehensive evaluation of structural conservatism of ribosomal proteins from evolutionarily distant organisms. Considering the current knowledge about features of the structural organization of the universal proteins and their intermolecular contacts, a possible role of individual proteins and their structural elements in the formation and functioning of ribosomes is discussed. The structural and functional conservatism of the majority of proteins of this group suggests that they should be present in the ribosome already in the early stages of its evolution.     

The basis of structural colour variation in two species of Papilio

An account is given of the ultrastructure and optical properties of the structural colour scales of Papilio karna and P.palinurus. The scales are Urania-type multilayers of 10 lamellae separated by air-spaces. A theoretical model is elaborated to account for the results and it is concluded that colour variation between species is achieved through control of film thickness. It is proposed that the surface structure functions as an anti-reflection coating. A generalised functional basis for the structural difference between Urania-type and Morpho-type scales is suggested.