A new view on Nematothallus: coralline red algae from the Silurian of Gotland

Abstract:  The thalloid carbonaceous fossil Nematothallus Lang, 1937, has been widely interpreted as an early Palaeozoic land‐plant, despite the absence of a convincing modern analogue. Exceptionally well‐preserved nematophyte cuticle from the Late Silurian Burgsvik Sandstone Formation, Gotland provides additional insight into the organism’s anatomy, phylogenetic affiliations and ecology. Because this material exhibits additional characters not present in the type material we assign it to Nematothallopsis gotlandii gen. et sp. nov. The organism was constituted of a close‐packed layer of palisade‐like filaments covered by a cuticle that bears a characteristic pseudocellular pattern on its inner surface. Apertures in this cuticle are often encircled by a ring of multicellular filaments, which are sometimes associated with spheroidal, spore‐like entities. In the light of the conspicuous similarity of the palisade layers to the pseudoparenchymatous tissue of coralline red algae, and of the filament‐fringed apertures to their reproductive conceptacles, we reconstruct the Nematothallopsis organism as an extinct rhodophyte and re‐evaluate the putative terrestrial habit of cuticular nematophytes in general.     

MORPHOLOGICAL REVERSION OF SPIRULINA (ARTHROSPIRA) PLATENSIS (CYANOPHYTA): FROM LINEAR TO HELICAL1

The cyanobacterium Spirulina Turpin is characterized by its regularly coiled trichomes. Under some conditions, its helical filaments can convert to abnormal morphologies, such as irregularly curved and even linear shapes, that had been considered as a permanent degeneration that could not be reversed. However, here we found that the linear filaments of Spirulina platensis Geitler could spontaneously revert to the helical form with the same morphology as the original filaments. Further studies showed that the ultrastructural, physiological, and biochemical characteristics of linear filaments were different from those of the original filaments, whereas they were the same for the reverted and the original filaments. The SDS‐PAGE analysis revealed at least four proteins or subunits related to Spirulina morphogenesis: The 21.9‐kDa and the 20.3‐kDa proteins were highly expressed in the helical filaments, whereas the 52.0‐kDa and the 31.8‐kDa proteins were highly expressed in the linear filaments. The random amplified polymorphic DNA analysis with 96 random primers showed that the genetic background of the reverted filaments was the same as that of the original filaments but distinct from that of the linear filaments. The results indicated that linear filaments of Spirulina could revert to the original morphology under certain conditions, and their other distinctive traits were regained.     

Filament formation inThermus species in the presence of some D-amino acids or glycine

A number of strains ofThermus spp. changed morphology from rods of about 6 to 8 m long to multicellular filaments (unsheathed trichomes) up to many hundreds of micrometres long with the addition of glycine or certain D-amino acids to the growth medium. Associated with this change was the formation of braided trichomes and occasionally true knots. Filament formation was reversible by the removal of the causal agent, but only if growth was possible. Electron microscopy suggested that the wall structure was not changed, but only that cells did not separate due to the continuous nature of the outer membrane layer. The filaments were thus multicellular. The constituent cells were similar in length to the normal rod-shaped cells. Filament formation byThermus spp. may have applications in industrial scale culture of these extracellular enzymeproducing thermophilic bacteria.     

Swimming behaviour of the multicellular magnetotactic prokaryote ‘Candidatus Magnetoglobus multicellularis’ under applied magnetic fields and ultraviolet light

Magnetotactic bacteria move by rotating their flagella and concomitantly are aligned to magnetic fields because they present magnetosomes, which are intracellular organelles composed by membrane-bound magnetic crystals. This results in magnetotaxis, which is swimming along magnetic field lines. Magnetotactic bacteria are morphologically diverse, including cocci, rods, spirilla and multicellular forms known as magnetotactic multicellular prokaryotes (MMPs). ‘Candidatus Magnetoglobus multicellularis’ is presently the best known MMP. Here we describe the helical trajectories performed by these microorganisms as they swim forward, as well as their response to UV light. We measured the radius of the trajectory, time period and translational velocity (velocity along the helix axis), which enabled the calculation of other trajectory parameters such as pitch, tangential velocity (velocity along the helix path), angular frequency, and theta angle (the angle between the helix path and the helix axis). The data revealed that ‘Ca. M. multicellularis’ swims along elongated helical trajectories with diameters approaching the diameter of the microorganism. In addition, we observed that ‘Ca. M. multicellularis’ responds to UV laser pulses by swimming backwards, returning to forward swimming several seconds after the UV laser pulse. UV light from a fluorescence microscope showed a similar effect. Thus, phototaxis is used in addition to magnetotaxis in this microorganism.     

COMPARATIVE DEVELOPMENT OF THE RED ALGAL PARASITES BOSTRYCHIOCOLAX AUSTRALIS GEN. ET SP. NOV. AND DAWSONIOCOLAX BOSTRYCHIAE (CHOREOCOLACACEAE,RHODOPHYTA)1

The development of two red algal parasites was examined in laboratory culture. The red algal parasite Bostrychiocolax australis gen. et sp. nov., from Australia, originally misidentified as Dawsoniocolax bostrychiae (Joly et Yamaguishi-Tomita) Joly et Yamaguishi-Tomita, completes its life history in 6 weeks on its host Bostrychia radicans (Montagne) Montagne. Initially the spores divide to form a small lenticular cell, and then a germ tube grows from the opposite pole. Upon contact with the host cuticle, the germ tube penetrates the host cell wall. The tip of the germ tube expands, and the spore cytoplasm moves into this expanded tip. The expanded germ tube tip becomes the first endophytic cell from which a parasite cell is cut off that fuses with a host tier cell. The nuclei of this infected host cell enlarge. As parasite development continues, other host-parasite cell fusions are formed, transferring more parasite nuclei into host cells. The erumpent colorless multicellular parasite develops externally on the host, and reproductive structures are visible within 2 weeks. Tetrasporangia are superficial and cruciately or tetra-hedrally divided. Spermatia are formed in clusters. The carpogonial branches are four-celled, and the carpogonium fuses directly with the auxiliary (support) cell. The mature carposporophyte has a large central fusion cell and sympodially branched gonimoblast filaments. Early stages of development differ markedly in Dawsoniocolax bostrychiae from Brazil. Upon contact with the host, the spore undergoes a nearly equal division, and a germ tube elongates from the more basal of the two spore cells, penetrates the host cell wall, and fuses with a host tier cell. Subsequent development involves enlargement of the original spore body and division to form a multicellular cushion, from which descending rhizoidal filaments form that fuse with underlying host cells. This radically different development is in marked contrast to the final reproductive morphology, which is similar to B. australis and has lead to taxonomic confusion between these two entities. The different spore germination patterns and early germ-ling development of B. australis and D. bostrychiae warrant the formation of a new genus for the Australian parasite.     

Sclerotin and lipid in the waterproofing of the insect cuticle

In all the cuticles studied waterproofing is effected by extracuticular material, a mixture of sclerotin precursors and lipids, exuded from the tubular filaments of the pore canals. In Rhodnius larval abdomen it is a layer of thickness similar to the outer epicuticle, believed to be composed of 'sclerotin' and wax, in Schistocerca larval sternal cuticle and in Carausius sternal cuticle it is similar. In Tenebrio adult sternal cuticle of the abdomen, in both the extracuticular exudation and the contents of the distal endings of the tubular filaments, the wax component is obscured by hard 'sclerotin'. In Manduca larva a very thin layer of 'sclerotin' and wax is covered by an irregular wax layer, average 0.75 micron, twice the thickness of the inner epicuticle. In Periplaneta and Blattella the abdominal cuticle is covered by a soft waxy layer, often about 1 micron thick, which is mixed with argentaffin material. Below this is a very thin waterproof layer of wax and 'sclerotin' continuous with the contents of the tubular filaments, which is readily removed by adsorptive dusts. In Apis adult abdominal terga free wax plus sclerotin precursors form a thin layer which is known to be removed by adsorptive dusts. In Calliphora larva there is a very thin layer of the usual mixed wax and sclerotin and below this a thick (0.5 micron) layer, lipid staining and strongly osmiophil, likewise extracuticular and exuded from the epicuticular channels. This material (which is often called 'outer epicuticle') has the same staining and resistance properties as the true outer epicuticle on which it rests. In the abdomen of Calliphora adult the waterproofing wax-sclerotin mixture forms a thin layer over the entire cuticle including the surface of the microtrichia. There is also a thin detachable layer of free wax on the surface.     

Enhancing the egg's natural defence against bacterial penetration by increasing cuticle deposition

The cuticle is a proteinaceous layer covering the avian egg and is believed to form a defence to microorganism ingress. In birds that lay eggs in challenging environments, the cuticle is thicker, suggesting evolutionary pressure; however, in poultry, selection pressure for this trait has been removed because of artificial incubation. This study aimed to quantify cuticle deposition and to estimate its genetic parameters and its role on trans‐shell penetration of bacteria. Additionally, cuticle proteins were characterised to establish whether alleles for these genes explained variation in deposition. A novel and reliable quantification was achieved using the difference in reflectance of the egg at 650 nm before and after staining with a specific dye. The heritability of this novel measurement was moderate (0.27), and bacteria penetration was dependent on the natural variation in cuticle deposition. Eggs with the best cuticle were never penetrated by bacteria (P < 0.001). The cuticle proteome consisted of six major proteins. A significant association was found between alleles of one of these protein genes, ovocleidin‐116 (MEPE), and cuticle deposition (P = 0.015) and also between alleles of estrogen receptor 1 (ESR1) gene and cuticle deposition (P = 0.008). With the heritability observed, genetic selection should be possible to increase cuticle deposition in commercial poultry, so reducing trans‐generational transmission of microorganisms and reversing the lack of selection pressure for this trait during recent domestication.     

Byssogenesis in the juvenile pink heelsplitter mussel,Potamilus alatus (Bivalvia: Unionidae)

The North American pink heelsplitter (Potamilus alatus) differs from most freshwater mussels in China by the ability to secrete an ephemeral byssus during its juvenile stage. In the present study, light microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) were used to investigate this ephemeral byssal structure, and amino acid composition was analyzed and compared with that of other species. The results revealed that the byssus consists of a long byssal thread and a few adhesive plaques which are randomly set up along the thread and assembled by petioles. There is a thin but distinctive cuticle with a continuous homogeneous matrix surrounding the byssal thread. Structural variation occurred when the byssal thread was differentially stretched. Four‐stranded helical primary fasciculi, which form a stable rope‐like structure, become evident after removal of the cuticle. The primary fasciculi consist of bundles of hundreds of parallel secondary fasciculi, each measuring about 5 μm in diameter. All evidence indicates that the byssus of the pink heelsplitter has a significantly different macrostructure and microstructure than the permanent byssus of the marine mussel Mytilus. Byssogenesis ceases when juveniles exceed 30 mm in length, although it varies greatly even among juveniles of similar size. Byssus formation is influenced by substrate type. The unique characteristics of the byssus have important advantages for survival, transition, and aggregation during the early life history. This study not only provides first insight into the structure of the ephemeral byssus and its relationship to freshwater mussel development and growth, but also suggests possibilities for the synthesis of novel biopolymer materials particularly useful in freshwater ecosystems. J. Morphol. 276:1273–1282, 2015. © 2015 Wiley Periodicals, Inc.     

Verosphacela silvae sp. nov. (Onslowiaceae, Phaeophyceae) from the Mediterranean Sea

We describe Verosphacela silvae sp. nov., from the Mediterranean Sea. It consists of horizontal filaments living on the lower face of the red alga Peyssonnelia rubra (Greville) J. Agardh, from which erect filaments up to 1.5 mm high rise and grow upright after passing through the thallus of the supporting species. There are both horizontal and erect filaments growing by apical cells. In the subapical cells, 1–2 longitudinal divisions occur (more frequently in the erect filaments) but no secondary transverse divisions occur. Erect filaments bear lateral propagules on a stalk of one to three (rarely more) cells. Propagules, with neither apical cells nor arms, consist of seven cells. Zoidangia are borne at the apex of erect laterals. The new species differs from V. ebrachia Henry mainly in habit, propagules and zoidangia. In addition, distinct from V. ebrachia, filaments of V. silvae never penetrate between the cuticle and the cell wall of the supporting alga. Moreover, propagules of V. silvae consist of seven cells, whereas those of V. ebrachia consist of 9–13 cells, and zoidangia are terminal on laterals in V. silvae, whereas in V. ebrachia they are sessile on both axes and laterals.     

Cutinsomes as building‐blocks of Arabidopsis thaliana embryo cuticle

Cutinsomes, spherical nanoparticles containing cutin mono‐ and oligomers, are engaged in cuticle formation. Earlier they were revealed to participate in cuticle biosynthesis in Solanum lycopersicum fruit and Ornithogalum umbellatum ovary epidermis. Here, transmission electron microscopy (TEM) and immunogold labeling with antibody against the cutinsomes were applied to aerial cotyledon epidermal cells of Arabidopsis thaliana mature embryos. TEM as well as gold particles conjugated with the cutinsome antibody revealed these structures in the cytoplasm, near the plasmalemma, in the cell wall and incorporated into the cuticle. Thus, the cutinsomes most probably are involved in the formation of A. thaliana embryo cuticle and this model plant is another species in which these specific structures participate in the building of cuticle in spite of the lack of the lipotubuloid metabolon. In addition, a mechanism of plant cuticle lipid biosynthesis based on current knowledge is proposed.     

Ultrastructural changes during reticulopod withdrawal in the foraminiferan protozoanAllogromia sp., strain NF

Summary Allogromia sp. is a benthic foraminiferan protozoan which extends and withdraws a dynamic network of branching and anastomosing pseudopodia,i.e., reticulopods. Each reticulopod contains an elongate cytoskeleton composed primarily of microtubules (MT). When withdrawal was induced with artificial seawater supplemented with MgCl₂, we found a time-dependent decrease in the number of reticulopodial MTs and a concomitant increase in 5-nm-diameter helical filaments. During the initial stages of withdrawal these helical filaments associated laterally to form loose aggregates. Later they formed dense paracrystalline aggregates, which appeared similar to those seen in the cell bodies of untreatedAllogromia juveniles prior to network extension. Similar results were obtained when withdrawal was induced by using seawater supplemented with other salts (NaCl, KCl). Treatment with an isotonic seawater substitute with an altered Na⁺:K⁺ ratio induced a momentary withdrawal, after which the organism recovered and reextended a network. During the withdrawal phase of this response, MTs became less abundant and aggregates of helical filaments more conspicuous. Together with earlier observations these findings suggest that helical filaments and paracrystalline material are an alternative or intermediate assembly form of MT proteins.     

Alzheimer disease paired helical filament core structures contain glycolipid.

The core structures of sodium dodecyl sulfate extracted, pronase digested paired helical filaments of Alzheimer disease were solubilized by heating in dimethyl sulfoxide. Electron microscopy revealed that after heating in dimethyl sulfoxide, intact paired helical filaments were no longer present in the dimethyl sulfoxide soluble fractions or in the insoluble lipofuscin-containing fractions. Enzyme-linked immunosorbent assays of the various fractions with the monospecific antibody A128 to paired helical filaments demonstrated 96% of the immunoreactivity to be in the dimethyl sulfoxide soluble fraction, and only 4% in the dimethyl sulfoxide insoluble fractions. Lyophilization of the dimethyl sulfoxide soluble supernatant and resuspension in water failed to reassociate the paired helical filaments, but did result in an insoluble precipitate. Analysis of the dimethyl sulfoxide solubilized paired helical filament fraction by nuclear magnetic resonance revealed it to be composed of glycolipid in a form that was distinct from similar fractions isolated from normal aged control brains. The aggregation of an altered glycolipid to form paired helical filaments in Alzheimer disease could explain their insolubility.     

Studies on ultrastructure and cultivation of microorganisms associated with zebra nematodes

Nematodes recovered from the hindgut of zebras were examined with scanning and transmission electron microscopy for microorganisms. Microorganisms were observed attached to the posterior extremities of two groups of nematodes, atractids and cyathostomes. Novel techniques were used to culture the microorganisms, and these included rinses to reduce contamination from hindgut flora and the design of the culture media. Electron microscopy revealed a flat bacterium not previously observed, as well as small rods and segmented filamentous bacteria. Culturing techniques resulted in isolation of a Propionibacterium species.Offprint requests to: R. C. Krecek.     

Accessory Pollen Adhesive from Glandular Trichomes on the Anthers of Leonurus sibiricus L. (Lamiaceae)

Abstract: Glandular trichomes (ca. 16 per anther) on the anthers of Leonurus sibiricus produce a secretion that, when touched, is liberated at once and becomes sticky when in contact with the air. With successive visitations of the pollinators (species of Bombus in naturalized populations) the number of secretion‐containing glands on each anther diminishes by mechanical rupture. On the pollinators, the secretion mixed with pollen was found adhered to the integument on the parts making contact with the anthers and stigma, mostly on the scape of the antennae. These trichomes are anatomically identical to the glandular scales common in the entire family and are formed by a multicellular cuticle‐bounded structure, with a foot and head. The secretion is accumulated as a milky emulsion under the cuticle, outside the primary cell wall, and is liberated by rupture of the cuticle. The composition possibly differs from what generally distinguishes these glandular trichomes, i.e. volatile oils that give these plants their particular smell. Such volatile compounds are generally assumed to have defensive or attractive functions, different from those observed in this study, which would be strictly mechanical.     

Hormonal control of growth and differentiation of insect tissues cultured in vitro

Summary This paper reviews the effects of insect hormones on lepidopteran imaginal discs cultured in vitro.β-ecdysone stimulated both evagination and cuticle deposition of wing discs ofPlodia interpunctella (Hübner). However, evagination required a shorter exposure to ecdysone than did cuticle deposition. Cuticle deposition was obtained under the following conditions: (a) a 24-hr pulse ofβ-ecdysone (0.5–5.0μg/ml); (b) continuous treatment with 0.2μg/mlβ-ecdysone; or (c) continuous treatment with 0.5 to 50.0μg/mlβ-ecdysone in medium conditioned with larval fat body. Investigations of some biochemical effects of ecdysone showed that RNA and protein synthesis was required for evagination and cuticle deposition. In particular, studies with actinomycin D and cycloheximide (at nontoxic levels) showed that RNA and protein synthesis during the ecdysone-dependent period was essential for subsequent development. These findings support the hypothesis that stimulation of macromolecular synthesis is fundamental to the action of ecdysone on imaginal discs. The influence of beta-ecdysone on chitin synthesis was also examined.β-ecdysone stimulated uptake and incorporation of tritiated-glucosamine by culturedP. interpunctella wing discs. Addition of hexosamines to the culture medium had no influence on ecdysone-induced cuticle deposition, but inhibition of glucose-uptake by cytochalasin B prevented the formation of cuticle. The action of ecdysone on particular enzymes in the chitin pathway remains to be elucidated. Presented in the formal symposium on Information Transfer in Eukaryotic Cells, at the 26th Annual Meeting of the Tissue Culture Association, Montreal, Quebec, June 2–5, 1975.     

Morphology, developmental ultrastructure and ultracytochemistry of staminal hairs in Bulbine inflata (Asphodelaceae) in relation to function

Results of light and electron microscopy and preliminary ultracytochemical studies of the staminal hairs of Bulbine inflata at different stages of development are reported here. The staminal filaments are covered with yellow, unicellular, linear, erecto-patent hairs. These staminal hairs arise directly as single cell outgrowths from epidermal cells of the filament. The surface of each hair is patterned with helical wall thickenings in an anticlockwise direction. This wall is covered by a thick folded cuticle, and formed of a loosely fibrillar cellulose layer. The hair cell possesses a cytoplasm rich in organelles. Especially ribosomes are abundant. Plastids contain large starch grains and peripheral lipid droplets. The smooth endoplasmic reticulum cisternae (SER) encircle the plastids and mitochondria; it is extended in the cytoplasm along the hair length. These hairs have functions in flower pollination attracting pollinators visually, secreting specific substances, providing increased surface area, protecting the filaments and being involved in their movement and vibration.     

A Nck‐associated protein 1‐like protein affects drought sensitivity by its involvement in leaf epidermal development and stomatal closure in rice

Water deficit is a major environmental threat affecting crop yields worldwide. In this study, a drought stress‐sensitive mutant drought sensitive 8 (ds8) was identified in rice (Oryza sativa L.). The DS8 gene was cloned using a map‐based approach. Further analysis revealed that DS8 encoded a Nck‐associated protein 1 (NAP1)‐like protein, a component of the SCAR/WAVE complex, which played a vital role in actin filament nucleation activity. The mutant exhibited changes in leaf cuticle development. Functional analysis revealed that the mutation of DS8 increased stomatal density and impaired stomatal closure activity. The distorted actin filaments in the mutant led to a defect in abscisic acid (ABA)‐mediated stomatal closure and increased ABA accumulation. All these resulted in excessive water loss in ds8 leaves. Notably, antisense transgenic lines also exhibited increased drought sensitivity, along with impaired stomatal closure and elevated ABA levels. These findings suggest that DS8 affects drought sensitivity by influencing actin filament activity.     

The Nature and Organization of Filaments in the Oral Apparatus of Tetrahymena1

ABSTRACT. Filaments in the oral apparatus of Tetrahymena appear similar, but not identical, to the intermediate filaments of multicellular organisms. The mean diameter of filaments measured in the present study was 16.4 nm, but the range of variation was much greater than has been reported for intermediate filaments. The organization of filaments within the oral apparatus has been studied by indirect immunofluorescence microscopy and immunogold localization at the electron microscopical level using antiserum raised in rabbits against the major subunit protein of the oral filaments (87K). The filaments were found to be organized into cables, networks, and chambers or cages which encase the basal bodies. At the highest level of organization, the filaments connect into a rigid framework capable of maintaining the overall architecture in the absence of microtubules. Like intermediate filaments, the oral filaments are insoluble at high ionic strength, have evolutionarily non-conservative subunit proteins, are probably non-contractile, and serve to stabilize persistent cellular architecture.     

A novel regulation on developmental gene expression of fruiting body formation in Myxobacteria

Myxobacteria are Gram-negative soil microorganisms that prey on other microorganisms. Myxobacteria have significant potential for applications in biotechnology because of their extraordinary ability to produce natural products such as secondary metabolites. Myxobacteria also stand out as model organisms for the study of cell–cell interactions and multicellular development during their complex life cycle. Cellular morphogenesis during multicellular development in myxobacteria is very similar to that in the eukaryotic soil amoebae. Recent studies have started uncovering molecular mechanisms directing the myxobacterial life cycle. We describe recent studies on signal transduction and gene expression during multicellular development in the myxobacterium Myxococcus xanthus. We provide our current model for signal transduction pathways mediated by a two-component His–Asp phosphorelay system and a Ser/Thr kinase cascade.