Skeletal Ultrastructure in some Tubuliporine Cyclostome Bryozoans

The ultrastructure of the calcareous skeleton is described in twenty–one species of recent tubuliporine cyclostome bryozoans, using field emission SEM. The succession of skeletal fabrics in interior walls may be classified into four different fabric suites. The first–formed part of the calcitic skeleton in all species for which it has been observed is a precursory fabric of tiny, wedge–shaped crystallites. This is succeeded in about half of the species studied by a fabric of transverse fibres, followed by foliated fabric and often semi–nacre (fabric suite 1). Most of the remaining species lack transverse fibres and have interior walls largely comprising semi–nacre (fabric suite 2). A few species have skeletons consisting of predominantly distally–oriented, irregularly or regularly foliated fabric (fabric suite 3). A single species has a skeleton of proximally–oriented foliated fabric (fabric suite 4). Basal exterior walls in all species have a precursory fabric of tiny wedge–shaped crystallites without a strong preferred orientation, deposited directly upon the organic cuticle, followed by a layer of planar spherulitic structure, which in turn is succeeded by a similar fabric to that developed in the interior wall of the species concerned. Outermost layers of frontal exterior walls exhibit one of the following combinations of three fabrics: an outer layer of (1) finely granular or wedge–shaped crystallites; a thin dense granular layer followed by (2) distally accreting planar spherulitic fabric., or (3) obliquely accreting planar spherulitic fabric growing partly towards the midline of the frontal wall. Terminal diaphragms usually have outer layers dominated by planar spherulitic ultrastructure with centripetal growth directions. The fabric suites present in tubuliporines encompass most known fabrics found in the other cyclostome suborders and support the notion that this species–rich suborder occupies a central position in cyclostome evolution.     

Skeletal ultrastructure and phylogeny of cyclostome bryozoans

Recent research on the ultrastructure of the calcareous skeleton in the bryozoan order Cyclostomata is summarized and updated, based on field emission SEM studies of 87 species. Six fundamental ultrastructural fabrics are recognized which differ in the crystallographic orientations, shapes and prevailing growth directions of the constituent crystallites. During the growth of individual walls a succession of fabrics is secreted, defining a fabric suite. Five fabric suites are described in interior walls and four in exterior walls. Nine ultrastructural characters were combined with 37 other skeletal characters in a PAUP analysis of the relationships between 28 post-Palaeozoic cyclostomes chosen to include representatives of all suborders. A single tree of length 142 steps was found. Comparison of tree statistics for three categories of characters showed ultrastructural characters to be more homoplastic than zooidal characters, and the latter more homoplastic than colonial characters. Rooting the tree on the paleotubuliporine Cuffeyella gave four transitions from fixed- to free-walled organization and no reversals. With respect to the five extant suborders of cyclostomes, this first, preliminary analysis implies that Rectangulata and Cancellata are monophyletic groups, whereas Articulata are diphyletic, and both Tubuliporina and Cerioporina paraphyletic.     

Skeletal Ultrastructures in some Cerioporine Cyclostome Bryozoans

Abstract The ultrastructure of the calcareous skeleton is described in nine species of Recent cyclostome bryozoans belonging to the suborder Cerioporina. Two species of Heteropora have interior zooecial walls comprising a granular precursory layer followed by a thick layer of transverse fibres and a subordinate foliated fabric with, in mature proximal walls, a semi-nacreous layer. The remaining seven species have interior walls with no transverse fibres and instead predominantly comprise a distally-imbricated, regularly foliated fabric overlying a granular precursory layer. Older, proximal surfaces often have abundant screw dislocations, but true semi-nacre is absent. Basal walls comprise an outer finely granular precursory fabric and planar spherulitic layer, succeeded by the same ultrastructural succession seen in the interior zooecial walls of the respective groups. Exterior walled diaphragms, peristomes and gonozooids similarly comprise an external fabric of planar spherulitic calcite, lined internally by the predominant fabric seen in the interior walls. Ultrastructurally, therefore, cerioporines may be split into two groups with different fabric suites, the first resembling cinctiporids and many tubuliporines in having interior walls with fabrics of transverse fibres, foliated crystallites and semi-nacre; and the second resembling the rectangulates Lichenopora and Disporella in having interior walls comprising only the foliated fabric. These findings support the close phylogenetic relationship between cerioporines and other cyclostomes but suggest that the cerioporines may constitute either a diphyletic or a paraphyletic group.     

Skeletal Ultrastructure in Some Articulate Cyclostome Bryozoans

The ultrastructure of the calcareous skeleton is described in 11 species of articulate cyclostome bryozoans with elastic joints. Ten species have interior walls comprising semi-nacreous and pseudofoliated fabrics without a precursory granular layer. Exterior walls consist of outer, finely granular and planar spherulitic layers, succeeded by semi-nacreous and pseudofoliated fabrics like those of interior walls. Outer fabrics are calcified as longitudinal strips, each corresponding to a planar sphcrulitic unit. Articulation surfaces comprise ring diaphragms of very fine granular fabric with concentric laminations. The semi-nacre of walls adjacent to ring diaphragms contains minute holes. Crisulipora occidentalis is unique in having interior walls of transverse fibres succeeded by pseudofoliated fabric, articulation surfaces festooned with deep pits but lacking well-differentiated ring diaphragms, and pseudopores containing sieve-like closure plates. The ultrastructure of most articulates resembles tubuliporine cyclostomes with dominantly semi-nacreous walls, although the lack of precursory granular fabric in the interior walls and the presence of subcircular tablets of semi-nacre (without six-fold sectoring) may be peculiar to articulates. In contrast, Crisulipora is more similar to other tubuliporines with transverse fibres. evidence which, together with other skeletal characters, suggests that Crisulipora evolved jointing independently of the rest of the articulate cyclostomes.     

Understanding the crystallographic and nanomechanical properties of bryozoans

This study examines how microscale differences in skeletal ultrastructure affect the crystallographic and nanomechanical properties of two related bryozoan species: (i) Hornera currieae, which is found at relatively quiescent depths of c. 1000 m, and (ii) Hornera robusta, which lives at depths of 50–400 m where it is exposed to currents and storm waves. Microstructural and Electron Backscatter Diffraction (EBSD) observations show that in both species the secondary walls are composed of low-Mg calcite crystallites that grow with their c-axes perpendicular to the wall. Branches in H. currieae develop a strong preferred orientation of the calcite c-axes, while in H. robusta the c-axes are more scattered. Microstructural observations suggest that the degree of scattering is controlled by the underlying morphology of the skeletons: in H. currieae the laminated branch walls are smooth and relatively uninterrupted, whereas the wall architecture of H. robusta is modified by numerous deflections, forming pustules and ridges associated with microscopic tubules. Modelling of the Young’s modulus and measurements of nanoindentation hardness indicate that the observed scattering of the crystallite c-axes affects the elastic modulus and nanohardness of the branches, and therefore controls the mechanical properties of the skeletal walls. At relatively high pressure in deep waters, the anisotropic skeletal architecture of H. currieae is aimed at concentrating elasticity normal to the skeleton wall. In comparison, in the relatively shallow and active hydrographic regime of the continental shelf, the elastically isotropic skeleton of H. robusta is designed to increase protection from external predators and stronger omni-directional currents.     

Skeletal Ultrastructure in the Cyclostome Bryozoan Hornera

Abstract Scanning electron microscopy of calcified walls in two species of the cyclostome bryozoan Hornera has revealed previously undescribed details of skeletal morphology and growth. The calcitic interior walls of both H. robusta MacGillivray and H. squamosa Hutton have a laminated structure. Walls are extended at distal growing edges where the formation of new crystallites is concentrated and wall fabric is nacreous or semi-nacreous. New crystallites are seeded on the surface of existing crystallites as six-sided rhombs. At the centres of the rhombs in H. robusta there are often three ‘spikes' which point towards alternate sides of the rhomb. Screw dislocations resulting in spiral overgrowths are also common at these distal wall edges. Wall thickening occurs further proximally where walls develop a regularly foliated structure of imbricated laths growing towards the colony base. Although often thought to be ubiquitous in cyclostomes, the division of walls into three layers (an inner, primary layer flanked on both sides by secondary layers) is absent in Hornera. Wall ultrastructure contrasts strongly with the lamellar–fibrous–lamellar structure recently described from cinctiporid cyclostomes. The c-axes of the crystallites are orientated perpendicular to the wall surface in Hornera, unlike cinctiporids in which they are orientated within the plane of the wall. Apparent similarities in ultrastructure suggest that Hornera may provide a good model for wall growth in extinct trepostome bryozoans.     

Ultrastructure and Development of the Ooecial Walls in Some Calloporid Bryozoans (Gymnolaemata: Cheilostomata)

In the brood chambers (ovicells) of six calloporid cheilostomes studied each skeletal wall consists of four calcified layers: (1) a very thin superficial layer of planar spherulitic crystallites, (2) an upper (outer) layer with wall-perpendicular prismatic ultrastructure, (3) an intermediate lamellar layer, and (4) a lower (inner) wall-perpendicular prismatic layer. Comparative studies of both the ovicell wall ultrastructure and early ovicell formation showed a hypothetical opportunity for evolving complex (multilayered) skeletal walls by fusion of the initially separated gymnocystal and cryptocystal calcifications in Cheilostomata. In two species studied, a bilobate pattern in the final stage of the formation of the ooecial roof was encountered in specimens with the cuticle preserved. A possible explanation to this finding is discussed – the bilobate pattern is suggestive of the hypothetical origin of the brood chamber from (1) two flattened spines, or (2) reduction in spine number of an originally multispinous ovicell.     

The body wall of cheilostome Bryozoa. I. The ectocyst of Watersipora nigra (Canu and Bassler)

The cuticle of Watersipora nigra is at first translucent, but it later becomes black and differentiates into two layers. It is composed, at least in part, of a protein-polysaccharide complex. Calcified parts are three-layered: (1) an outer, cuticular layer, (2) a calcium carbonate skeleton deposited on a matrix of acid mucopolysaccharide, and (3) a “skeletal membrane.” The relationships of these layers indicate that the skeleton is intracuticular. A layer of cuticular material, the “intercalary cuticle” is present in lateral walls, but not transverse walls; it may become calcified in some species. The cuticles of calcified and uncalcified parts of cheilostomes are not necessarily homologous.     

Multizooidal Budding in Parasmittina trispinosa (Johnston) (Bryozoa,Cheilostomata)

Two principally different wall types occur in the bryozoan colony: Exterior walls delimiting the super-individual, the colony, against its surroundings and interior walls dividing the body cavity of the colony thus defined into units which develop into sub-individuals, the zooids. In the gymnolaemate bryozoans generally, whether uniserial or multiserial, the longitudinal zooid walls are exterior, the transverse (proximal and distal) zooid walls interior ones. The radiating zooid rows grow apically to form “tubes” each surrounded by exterior walls but subdivided by interior (transverse) walls. The stenolaemate bryozoans show a contrasting mode of growth in which the colony swells in the distal direction to form one confluent cavity surrounded by an exterior wall but internally subdivided into zooids by interior walls. In the otherwise typical gymnolaemate Parasmittina trispinosa the growing edge is composed of a series of “giant buds” each surrounded by exterior walls on its lateral, frontal, basal and distal sides and forming an undifferentiated chamber usually 2–3 times as broad and 3 or more times as long as the final zooid. Its lumen is subdivided by interior walls into zooids 2–3, occasionally 4, in breadth. This type of zooid formation is therefore similar to the “common bud” or, better-named, “multizooidal budding” characteristic of the stenoleamates but has certainly evolved independently as a special modification of the usual gymnolaemate budding.     

A set of possible sensory system preserved in cuticle of Late Devonian thylacocephalan arthropods from Poland

The cuticle of concavicarids (Arthropoda: Thylacocephala) from the early Famennian (Upper Devonian) of the Holy Cross Mountains, Poland was studied with respect to its microstructural details. Investigated laminated cuticle with phosphatic/organic composition, possesses two different kinds of microstructure with assumed sensory functions. The first kind consists of circular depressions, each located in the central area of characteristic polygons forming the carapace exterior ornamentation. These depressions are interpreted as sealed during phosphatisation processes setal lumens. The second kind occurs exclusively in carapace margins. These intracuticular microstructures occur as elongated tubular structures, circular to oval in cross section, penetrating the cuticle interior but not reaching its surface. They form a thin belt with a kind of ‘sensory fields’ on the dorsum and a wider belt in the ventrocaudal part of the carapace. These belts are connected in the rostral and caudal area, forming a continuous sensory zone encompassing each valve. These structures are very similar to crustacean organule canals, and the dorsally situated ‘sensory fields’, suggest some similarity to crustacean sensory dorsal organs. This possible sensory system is the oldest of this kind found in Thylacocephala. Its morphology and presumed canal walls mineralogical composition suggests crustacean affinity of Thylacocephala.     

The presence of cutan limits the interpretation of cuticular chemistry and structure: Ficus elastica leaf as an example

Plant cuticles have been traditionally classified on the basis of their ultrastructure, with certain chemical composition assumptions. However, the nature of the plant cuticle may be misinterpreted in the prevailing model, which was established more than 150 years ago. Using the adaxial leaf cuticle of Ficus elastica, a study was conducted with the aim of analyzing cuticular ultrastructure, chemical composition and the potential relationship between structure and chemistry. Gradual chemical extractions and diverse analytical and microscopic techniques were performed on isolated leaf cuticles of two different stages of development (i.e. young and mature leaves). Evidence for the presence of cutan in F. elastica leaf cuticles has been gained after chemical treatments and tissue analysis by infrared spectroscopy and electron microscopy. Significant calcium, boron and silicon concentrations were also measured in the cuticle of this species. Such mineral elements which are often found in plant cell walls may play a structural role and their presence in isolated cuticles further supports the interpretation of the cuticle as the most external region of the epidermal cell wall. The complex and heterogeneous nature of the cuticle, and constraints associated with current analytical procedures may limit the chance for establishing a relationship between cuticle chemical composition and structure also in relation to organ ontogeny.     

A putative new hydrostatic skeletal function for the epidermis in Monhysterids (Nematoda)

The ultrastructure of the epidermis of two Monhysterid nematodes (Geomonhystera disjuncta and Diplolaimella dievengatensis) is studied in detail. The epidermis is composed of discrete uninucleated cells. The cytoplasmic layer of the epidermis between the cuticle and the somatic muscles is very thin and contains bundles of filaments that attach the muscles to the cuticle. The epidermal chords are voluminous and contain the nuclei and most of the cell organelles. In the chords many large electron-transparent vacuoles are found. It is hypothesized that these vacuoles fulfill a function as a compartmentalised hydrostatic skeleton.     

The body wall of cheilostome Bryozoa. II. Interzoidal communication organs

Communication organs (septulae) of cheilostome Bryozoa are more complex than perviously believed. Annuli, present only in lateral septulae, are thickenings of the intercalary cuticle. Each communication pore is filled with a ring-like “pore cincture,” through which project a pair of “special cells.” Septulae of all species examined (10 species from 6 families) can be considered modifications of the same structure, varying only in degree of calcification and number of communication pores. External walls, including basal and lateral walls, are best defined as reinforcements of the ectocyst, which is derived by intussusception from the primary cuticle of the ancestrula. The lateral ectocyst must be considered a double layer formed by invagination of the distal ectocyst. Internal walls are developed by apposition from inner parts of the ectocyst; they include pore plates and transverse walls. External walls are laid down first. Lenticular masses develop unilaterally on the uncalcified lateral ectocyst; the pore plate develops by apposition from the interior part of the ectocyst. Depending on the species, the pore plate may or may not be calcified at the time of its formation. Communication pores are formed when the developing pore plate abuts against embryonic special cells. The septular ectocyst never calcifies; it breaks down when the pore plate is complete. Some ascophorans undergo “reparative budding,” in which new zoids are formed within dead zoecia. Hollow, ectocyst-covered buds lined with blastemic epithelia are produced from septulae of live zoids; adjacent buds may fuse. These findings are consistent with the view that lateral septulae are aborted zoids and that pore plates represent transverse walls.     

Atomic force microscopy of microfibrils in primary cell walls

Examination of angiosperm primary cell walls by transmission electron microscopy shows that they contain microfibrils that probably consist of cellulose microfibrils surrounded by associated non-cellulosic polysaccharides. Previous studies using solid-state (13)C NMR spectroscopy have shown that the cellulose is all crystalline with crystallites of cross-sectional dimensions of 2-3 nm. However, it is not known if each microfibril contains only one, or more than one crystallite because there is no agreement about the dimensions of the microfibrils. Partially hydrated primary cell walls isolated from onion ( Allium cepa L.) and Arabidopsis thaliana (L.) Heynh. were examined by atomic force microscopy and the microfibril diameters determined. The cell walls of both species contained tightly interwoven microfibrils of uniform diameter: 4.4+/-0.13 nm in the onion and 5.8+/-0.17 nm in A. thaliana. The effect was also examined of extracting the A. thaliana cell walls to remove pectic polysaccharides. The microfibrils in the extracted cell walls of A. thaliana were significantly narrower (3.2+/-0.13 nm) than those in untreated walls. The results are consistent with the microfibrils containing only one cellulose crystallite.     

Periclinal penetration of potassium permanganate into mature cuticular membranes ofAgave andClivia leaves: new implications for plant cuticle development

Staining cuticular membranes ofAgave americana andClivia miniata en bloc with potassium permanganate results in a strong contrast in the interior cuticular layer while the exterior part remains unstained. This is not caused by a selective chemical reaction with the interior part but by the unidirectional penetration of the reagent from the interior side, the outside being protected by the cuticle proper. In transverse cryosections of the cuticular membrane, permanganate penetrates nearly as easily into the exterior cuticular layer as into the interior one giving the same contrast. However, compared with the periclinal penetration into the cuticle proper this penetration is accelerated five-to tenfold by the polysaccharide network within the cuticular layer which serves as a distribution-channel system. Periclinal penetration into the cuticle proper occurs independently in each cutin penetration unit included between two obvious lucent lamellae and further divided into subunits.     

Systematics and phylogeny of the cheilostome bryozoan Doryporella

The four nominal species of the Arctic-Boreal anascan bryozoan Doryporella Norman are revised. Colonies of Doryporella are small, typically encrust shells and pebbles, and have autozooids with frontal walls bearing a distinctive reticulate pattern of polygonal ridges. High resolution SEM shows the frontal wall to be an interior wall (cryptocyst), contrary to previous studies which have regarded it as an exterior wall (gymnocyst). The inter-relationships between the species assigned to Doryporella are assessed cladistically using skeletal characters. The type species ( D. spathulifera ) and two other species ( D. alcicornis and D. armata ) form a well-supported clade, but the fourth species ( D. reticulata ) is more closely-related to certain other calloporid genera. Therefore, the reticulate frontal wall is inferred to have evolved twice and D. reticulata is removed to Doryporellina gen. n. Biogeographical distributions of the four species are reviewed.     

Studies on the Ultrastructure and Histochemistry of Plant Cuticles: The Cuticular Membrane of Agave americana L. in situ

The outer epidermal wall of Agave americana leaves was examinedin order to gain more information about the location and chemicalconstitution of the structural components. In middle aged leavesthe wall comprised six layers which were designated epicuticularwax, cuticle proper, exterior and interior cuticular layer,exterior and interior cellin wall. A lamellated structure, consistingof a series of electron translucent lamellae of uniform thicknessalternating with opaque ones of variable thickness, was observedin the thin cuticle proper on the outside of the cuticular membrane,even without heavy metal treatment. The cuticular layers underneathformed the bulk of the cuticular membrane and they also hadtwo components, an amorphous matrix permeated by a reticulumof fibrillae. Cutin, detected with osmium and with iodine/iodine-sulphuricacid–silver proteinate, was a major component of the opaquelamellae of the cuticle proper and the matrix of the cuticularlayer. Carbohydrates were absent from the cuticle proper butwere detected specifically in the fibrillae of the cuticularlayer and in the cellin wall. Pectic material seemed to be presenton both sides of the junction between cuticular membrane andcellin wall, but no discrete zone corresponding to light microscopicalobservations was detected in the electron microscope. Althoughthe lucent lamellae of the cuticle proper were tentatively ascribedto wax there was no structural or ultrahistochemical evidencefor the wax component of the cuticular layer. The various ultrahistochemicalreactions are discussed in relation to the known chemical compositionof the membrane. Agave americana L., epidermis wall, cuticular membrane, cuticle proper, cuticular layer, ultrahistochemistry, wax     

Cytological Investigations of Lithobiontic Microorganisms in Granitic Rocks

This paper shows the ultrastructure of lithobiontic organisms in a granitic rock from the exterior to the interior, where fissures are found 1–2 mm from the surface. There is clear differentiation at cell level between mycobiont and photobiont cells located on the rock surface which forms part of the lichen thallus and prokaryotic and eukaryotic cells (algae and fungi) found in the fissures. As well as the observations at the ultrastructural level of the microorganisms which live in fissures and cavities, immunolabelling techniques with colloidal gold have been applied to obtain an immunolocalization of Rubisco enzyme in some of the cells. The technique applied here permits Rubisco enzyme to be identified in algae-like cells belonging to fissures where it is difficult to identify the pyrenoid. The mineral environment of the cells situated inside the fissures is investigated by Energy Dispersive Spectrometry (EDS). The biotite particles present in the fissure walls, as well as some lithobiont microorganisms, show a depletion of potassium from interlayer positions.     

A new early cyclostome bryozoan from the Lower Ordovician (Volkhov Stage) of Russia

Goryunovia hemiseptata n.gen. n.sp. is an unusual Lower Ordovician cyclostome bryozoan, represented by a single colony from the Middle Volkhov Stage (Arenig) of the Leningrad Oblast. The encrusting uniserial colony consists of large, slender zooids, many with closely-spaced partial diaphragms (hemisepta), and has a distinctive trichotomous branching pattern. Skeletal organization is of the sagenellid grade: calcified interior walls appear to be present, and the exterior frontal walls lack pseudopores. As details of microstructure and early astogeny are unknown, it is difficult to interpret the phylogenetic relationships ofGoryunovia until further material is forthcoming. However, there is some resemblance with the enigmatic hederellids, which appear in the Upper Silurian and possibly range into the Triassic.     

新疆亚鳞木（比较种）角质层特征

报道了产自江苏宜兴晚泥盆世五通组新疆亚鳞木（比较种）Ｓｕｂｌｅｐｉｄｏｄｅｎｄｒｏｎ ｃｆ． ｘｉｎｊｉａｎｇｅｎｓｅＳｕｎ）的表皮角质层用荧光分析显示的特征．该种茎干表皮角质层覆于叶座及叶座间隔带,其中间隔带角质层厚于叶座．表皮细胞在间隔带与叶座表现特征不同．间隔带中部,表皮细胞呈纵长的多边形,其纵长方向与茎干延伸方向相同,细胞壁略有弯曲．间隔带靠近叶座之表皮细胞,细胞壁直,形状类似于前者;但大小仅为前者的１／２左右,且其纵长方向逐渐向叶座边缘偏转．叶座表皮细胞呈近等多边形,有胞间隙．该种茎表皮无气孔