Eukaryotic picoplankton, Mychonastes homosphaera (Chlorophyceae, Chlorophyta), in Lake Kinneret, Israel

Eukaryotic picoplankton was isolated from Lake Kinneret, Israel. This green picoplankton is spheroidal or ovoidal in shape. The cell wall has a network of ribs on its surface. The chloroplast is cup- or mantel-shaped without a pyrenoid. The morphology and cellular structure of this autosporic alga is identical with Mychonastes homosphaera (Skuja) Kalina et Pun?ochá?ová. The phylogenetic tree based on 18S ribo-somal RNA gene sequence data showed that M. homosphaera is placed in the Chlorophyceae and located at the base of a branch leading to the other algae possessing directly opposed orientation of the flagellar apparatus.     

Recent progress in Bacillus subtilis spore-surface display: concept,progress, and future

With the increased knowledge on spore structure and advances in biotechnology engineering, the newly developed spore-surface display system confers several inherent advantages over other microbial cell-surface display systems including enhanced stability and high safety. Bacillus subtilis is the most commonly used Bacillus species for spore-surface display. The expression of heterologous antigen or protein on the surface of B. subtilis spores has now been practiced for over a decade with noteworthy success. As an update and supplement to other previous reviews, we comprehensively summarize recent studies in the B. subtilis spore-surface display technique. We focus on its benefits as well as the critical factors affecting its display efficiency and offer suggestions for the future success of this field.

     

An extraordinary new carnivorous sponge,Chondrocladia lyra,in the new subgenus Symmetrocladia (Demospongiae,Cladorhizidae), from off of northern California,USA

Chondrocladia (Symmetrocladia) lyra subgen. nov., sp. nov., is described from northeast Pacific sites at Escanaba Ridge and Monterey Canyon at depths of 3316–3399 m. Two retrieved specimens are described in detail, while variations are described in ten photographed or videotaped specimens. The basic structure, termed a vane, is harp‐ or lyre‐shaped. From 1 to 6 vanes extend by radial growth from the organism's center. The orientation among the vanes is approximately equiangular, such that together they display pentaradiate, tetraradiate, triradiate, or biradiate symmetries. Each vane is formed by a horizontal stolon supporting a series of upright, equidistantly spaced branches each of which terminates at its apex in a swollen ball in all observed specimens except the paratype. Swellings occur midway along the branches in the holotype, but not in the paratype. A linear row of filaments project from the sides, front, and back of each branch, and also from the tops of each stolon. The terminal balls are the sites of spermatophore production and release; mid‐branch swellings are sites of oocyte maturation. The two megasclere spicule types have specific distributions; styles support rhizoids, stolons, and branches, while subtylostyles support filaments and terminal balls. Anchorate isochelae cover all surfaces. Enclosed crustacean prey on branches and stolons provide direct evidence of carnivory. The structure of the vanes maximizes surface area for passive suspension feeding. Increased surface area could also maximize spermatophore capture, with the sigmas projecting from the spermatophore surface being caught by projecting isochelae on filaments. Swellings on filaments are snared spermatophores, firmly fused to recipient tissues and undergoing destruction. Spermatophores on filaments are present in branch swellings containing early and mature oocytes. Oogenesis and maturation occur only in proximity to branch swellings, suggesting that development is induced by spermatophore reception. Symmetrical development of uniserial branched stolons (the vanes) characterized members of the new subgenus Symmetrocladia.     

PrgB promotes aggregation,biofilm formation,and conjugation through DNA binding and compaction

Gram‐positive bacteria deploy type IV secretion systems (T4SSs) to facilitate horizontal gene transfer. The T4SSs of Gram‐positive bacteria rely on surface adhesins as opposed to conjugative pili to facilitate mating. Enterococcus faecalis PrgB is a surface adhesin that promotes mating pair formation and robust biofilm development in an extracellular DNA (eDNA) dependent manner. Here, we report the structure of the adhesin domain of PrgB. The adhesin domain binds and compacts DNA in vitro. In vivo PrgB deleted of its adhesin domain does not support cellular aggregation, biofilm development and conjugative DNA transfer. PrgB also binds lipoteichoic acid (LTA), which competes with DNA binding. We propose that PrgB binding and compaction of eDNA facilitates cell aggregation and plays an important role in establishment of early biofilms in mono‐ or polyspecies settings. Within these biofilms, PrgB mediates formation and stabilization of direct cell‐cell contacts through alternative binding of cell‐bound LTA, which in turn promotes establishment of productive mating junctions and efficient intra‐ or inter‐species T4SS‐mediated gene transfer.     

Expression and Characterization of Fibroblast Growth Factor 8 from Mexican Axolotl, Ambystoma mexicanum

Fibroblast growth factor (FGF) has been known to regulate the proliferation and differentiation of a variety of cell types via interaction with a specific FGF receptor on the cell surface. In the present study, Fgf8 cDNA of Mexican axolotl, Ambystoma mexicanum, was expressed in Escherichia coli as an MBP-FGF8 fusion protein. The cell proliferation activity of the recombinant FGF8 (rFGF8) was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazoliumbromide (MTT) assay. The addition of rFGF8 to the culture medium enhanced proliferation of BALB/c 3T3 and BHK21 cells about 1.4--1.5 fold. To analyze the binding activity of rFGF8 to the cell surface, cell surface enzyme linked immunosorbent assay was developed. Comparison of the structure of basic FGF with the computer-simulated structure of FGF8 suggested that Tyr-58, Glu-132, Tyr-139, and Leu-179 might be the potential receptor binding sites. Amino acid substitution muteins of FGF8 were constructed by PCR-derived directed mutagenesis and the muteins were overexpressed in E. coli. The rFGF8 muteins were purified and their binding activities were analyzed. Substitution of Tyr-58 or Glu-132 or Leu-179 of the FGF8 with alanine reduced the binding affinity, while substitution of Tyr-139 with alanine did not alter the binding affinity. These results imply that Tyr-58, Glu-132, and Leu-179 of FGF8 might be involved in its binding to the cell surface.     

Ni‐Metalloid (B,Si, P,As, and Te) Alloys as Water Oxidation Electrocatalysts

Breakthroughs toward effective water‐splitting electrocatalysts for mass hydrogen production will necessitate material design strategies based on unexplored material chemistries. Herein, Ni‐metalloid (B, Si, P, As, Te) alloys are reported as an emergent class of highly promising electrocatalysts for the oxygen evolution reaction (OER) and insight is offered into the origin of activity enhancement on the premise of the surface electronic structure, the OER activation energy, influence of the guest metalloid elements on the lattice structure of the host metal (Ni), and surface‐oxidized metalloid oxoanions. The metalloids modify the lattice structure of Ni, causing changes in the nearest Ni–Ni interatomic distance (d_Ni–Ni). The activation energy E_a scales with d_Ni–Ni indicating an apparent dependence of the OER activity on lattice properties. During the OER, surface Ni atoms are oxidized to nickel oxyhydroxide, which is the active state of the catalyst, meanwhile, the surface metalloids are oxidized to the corresponding oxoanions that affect the interfacial electrode/electrolyte properties and hence the adsorption/desorption interaction energies of the reacting species.     

Ultrastructural features of the egg envelope of silver carp, <Emphasis Type="Italic">Hypophthalmichthys molitrix</Emphasis> (Osteichthyes,Cyprinidae)

Synopsis We investigated the ultrastructure of the egg membrane surface (unfertilized egg) of Hypophthalmichthys molitrix using scanning electron microscopy. The eggs of silver carp like most other teleosts are surrounded by a relatively thick egg membrane, and have a type III micropyle at the animal pole. The micropyle is almost circular in shape and the micropyle canal is located in its center. The micropyle region is not flat. Round or oval accessory pores are also observed in the canal. The surface of zona radiata is wavy and uneven with a uniform distribution of almost round pores with lips. Microvilli like structure are found in the pore opening region.     

Embryonic development of Zoraptera with special reference to external morphology,and its phylogenetic implications (Insecta)

The embryonic development of Zorotypus caudelli Karny (Zoraptera) is described with the main focus on its external features. A small heart‐shaped embryo is formed on the dorsal side of the egg by the fusion of paired blastoderm regions with higher cellular density. The orientation of its anteroposterior axis is opposed to that of the egg. This unusual condition shows the potential autapomorphy of Zoraptera. The embryo extends along the egg surface and after reaching its full length, it migrates into the yolk. After developing there for a period of time, it reappears on the surface, accompanied by a reversion of its anteroposterior axis, finally taking its position on the ventral side of the egg. The definitive dorsal closure completes, and the prelarva hatches after perforating the chorion with very long egg tooth formed on the embryonic cuticle. Embryological data suggest the placement of Zoraptera among the “lower neopteran” or polyneopteran lineage: features supporting this are embryo formation by the fusion of paired regions with higher cellular density and blastokinesis accompanied by full elongation of the embryo on the egg surface. The extraordinarily long egg tooth has potential synapomorphy with Embioptera or Eukinolabia (= Embioptera + Phasmatodea). Together with the results from our previous studies on the egg structure, male reproductive system and spermatozoa, the close affinity of Zoraptera with Eukinolabia appears likely, that is, a clade Zoraptera + (Embioptera + Phasmatodea). J. Morphol. 275:295–312, 2014. © 2013 Wiley Periodicals, Inc.     

Morphology,ultrastructure, and development of the parasitic larva and its surrounding trophamnion of Polypodium hydriforme Ussov (Coelenterata)

Summary The larval stage of Polypodium hydriforme is planuliform and parasitic inside the growing oocytes of acipenserid fishes. The larva has inverted germ layers and a special envelope, the trophamnion, surrounding it within the host oocyte. The trophamnion is a giant unicellular provisory structure derived from the second polar body and performing both protective and digestive functions, clearly a result of adaptation to parasitism. The trophamnion displays microvilli on its inner surface, and irregular protrusions anchoring it to the yolk on its outer surface. Its cytoplasm contains long nuclear fragments, ribosomes, mitochondria, microtubules, microfilaments, prominent Golgi bodies, primary lysosomes, and secondary lysosomes with partially digested inclusions.The cells of the larva proper are poorly differentiated. No muscular, glandular, neural, interstitial, or nematocyst-forming cells have been found. The entodermal (outer layer) cells bear flagella and contain rough endoplasmic reticulum; the ectodermal (inner layer) cells lack cilia and contain an apical layer of acid mucopolysaccharid granules. The cells of both layers contain mitochondria, microtubules, and Golgi bodies; their nuclei display large nucleoli with nucleolonema-like structure, decondensed chromatin, and some perichromatin granules. At their apical rims, the ectodermal cells form septate junctions; laterally, the cells of both layers form simple contacts and occasional interdigitations. The lateral surfaces of entodermal cells are strengthened by microtubules.     

Phylogeny and Establishment of a New Ciliate Family,Wilbertomorphidae fam. nov. (Ciliophora,Karyorelictea), a Highly Specialized Taxon Represented by Wilbertomorpha colpoda gen. nov., spec. nov.

The morphology and molecular phylogeny of a new marine interstitial karyorelictean ciliate, Wilbertomorpha colpoda gen. nov., spec. nov., isolated from a sandy beach at Qingdao, China, are investigated. The ciliature is mainly on the right surface with the left surface barren except for a single marginal kinety, which is typical of the order Loxodida. Nevertheless, other morphological features, such as the highly reduced buccal structure, the absence of a buccal cavity and the presence of extremely specialized cortical granules, prevent its assignation to any known family. Furthermore, the secondary structure of the variable region 2 of the SSU rRNA gene is unique among loxodids. Thus, we establish a new family, Wilbertomorphidae fam. nov. for the genus Wilbertomorpha. Phylogenetic analyses based on SSU rRNA gene sequence data suggest that this taxon is a sister group to the family Geleiidae which belongs to the order Protoheterotrichida. However, morphologically Wilbertomorpha bears a much stronger resemblance to Loxodidae which belongs to the order Loxodida. Based on these findings, we tentatively place Wilbertomorphidae nov. fam. incertae sedis within Karyorelictea.     

Effect of Ultrasonic Treatment on Structure,Antibacterial Activity of Sugarcane Leaf Polysaccharides

This study investigated the impact of ultrasonic extraction (UE) on the structure and in vitro antibacterial activity of polysaccharides from sugarcane leaves (SLW). Native sugarcane leaf polysaccharides were treated with ultrasound (480 W) for 3 h to yield sugarcane leaf polysaccharides (SLU). Compared to SLW (33.59 kDa), the molecular weight of SLU (13.08 kDa) was significantly decreased, while the monosaccharide composition of SLU was unchanged. The results of SEM and XRD indicated that UE significantly changed the surface morphology of SLW and destroyed its inner crystalline structure. In vitro experiments showed that SLU had stronger antibacterial activity. These findings revealed that UE treatment could alter the tertiary structure of SLW but had no impact on its primary structure. Furthermore, the antibacterial activity of SLW could be greatly enhanced after UE treatment. As a bioactive additive, SLU has great application potential in functional foods, cosmetics, and pharmaceuticals.     

STRUCTURE OF THE ROYAL ANN CHERRY CUTICLE AND ITS SIGNIFICANCE TO CUTICULAR PENETRATION

The structure of the Royal Ann cherry cuticle (Prunus avium L.) was determined and interpreted in the light of its possible significance to cuticular penetration by a SO₂-calcium bisulfite brine. The morphology of the cuticle was determined by standard histological and histochemical techniques. The surface structure of the cuticle was found to have a smooth to granular sheet or layer of surface wax, which when removed revealed a porous sponge-like surface. The cuticular surface showed intermittent birefringence, which increased as the fruit matured. Ectodesmata were found to occur over anticlinal walls and in guard cells on both sides of the fruit, with more on the side opposite the suture. Both sides had stomata with more occurring on the suture side. Secondary bleaching was found to alter the structure and permeability of the cuticle.     

Cauda epididymal spermatozoa of the rufous hare wallaby, Lagorchestes hirsutus (Metatheria, Mammalia) imaged by electron and confocal microscopy

The ultrastructure of mature Lagorchestes hirsutus spermatozoa is described for the first time, revealing unusual aspects of sperm structure in macropodid species. The sperm head is ovoid rather than cuneiform, lacks a ventral nuclear groove and has an acrosomal distribution over approximately 85–90% of its dorsal surface. Immediately adjacent to the nuclear membrane the peripheral nucleoplasm in most spermatozoa form an irregular series of distinctive evaginations previously not described in the spermatozoa of any other marsupial. The midpiece is extremely thickened and short, containing no helical network or peripheral plasma membrane specializations. Axonemal structure is unspecialized with no connecting lamellae; dense outer fibres are closely adherent to axonemal doublets. The sperm morphology of this species is highly aberrant in comparison to other macropod taxa and supports the retention of Lagorchestes as a distinctive genus. In light of this new information, skeletal and serological data should be re‐evaluated to determine the true taxonomic and phylogenetic position of this species.     

Durability of Simuliid Silk Pads (Simuliidae,Diptera)

A central aspect of simuliid adaption to life in moving water is a silky secretion which they produce in their salivary glands. This secretion is spread on the substrate surface thereby enabling adhesion of the posterior abdominal hooks of the larvae to the substrate surface. Therefore, silk pads are a prerequisit for simuliid filter‐feeding as well as any kind of locomotive activity. If silk pad adhesion is weak, larvae risk drifting off, either directly or during locomotion. Properties of the adhesive (=silk) as well as the substrate surface may cause weak adhesion. A specialist such as S. noelleri which has little chance of surviving after drifting off its lake outlet habitat, should have adaptations to reduce this risk. Such an adaptation could be very durable silk pads, giving larvae the chance to be safely attached for a relatively long time. In this study larval silk pads of S. ornatum and S. noelleri were stained using Giemsa's staining procedures. Changes in silk pad structure and traces of rot after 2–33 days of exposure to stream water were recorded and compared. Silk pads of S. ornatum and S. noelleri showed differences which indicate differences in ageing processes and biochemistry of this secretion.     

Variation on the population,racial, and species level in the primitive relic Angiosperm genusDrimys (Winteraceae) in South America

Statistical analyses and scatter diagrams illustrate for the polymorphic E. and SE. BrazilianDrimys brasiliensis a clear correlation between morphological variation and eco-geographical differentiation. This is backed by data on the (ultra)structure of the lower leaf surface. A new infraspecific taxonomy is proposed, andD. roraimensis andD. angustifolia are recognized as distinct species of the Guyana Highlands and S. Brazil, respectively. The affinities between these and other taxa of the paleopolyploid genus, its evolution and its position within the family are discussed.Dedicated to Prof.Lothar Geitler at the occasion of his 80th birthday.     

Genetically separate populations of the ocean‐skater Halobates sericeus (Heteroptera: Gerridae) have been maintained since the late Pleistocene

The oceanic water strider (or ocean‐skater) Halobates sericeus Eschscholtz has a disjunct distribution in the Pacific Ocean, with northern and southern populations widely separated by an equatorial zone. It is sensitive to sea surface conditions and, consequently, its distribution and population structure may provide an insight into environmental changes on the ocean surface on both recent and historical time scales. We assessed the genetic diversity and population structure of H. sericeus in the Pacific Ocean using three gene markers – cytochrome oxidase subunit I (COI), elongation factor 1α and internal transcribed spacer 1 (ITS‐1). These markers indicate that both populations are evolutionarily distinct with limited gene flow, having separated 20 000–50 000 years ago. This suggests that physical conditions and/or biotic interactions on the surface of the Pacific Ocean have provided significant barriers to gene flow since the late Pleistocene or earlier, creating biotic stability over large geographical and temporal scales in spite of a long history of global climate change. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 797–805.     

Behavior,activities, and effects of bacteria on synthetic quartz monocrystal surfaces

Two strains ofBacillus sp. and a strain ofBrevibacterium sp., originally isolated from a natural quartzite surface, were characterized and employed as test strains with several methods: acridine orange fluorochromation and epifluorescence microscopy were used for detection of individual cells; scanning and transmission microscopy for studying attachment behavior; replica techniques in combination with electron microscopy for following surface interaction effects; and chemical analysis of SiO₂ for detecting possible silica leaching activities. The experimental results clearly showed that the three test strains were able to attach to and grow on the precleaned quartz surfaces. Attachment modes were either by direct sorption mechanisms (Brevibacterium sp. S) or the production of adhesive polymers (Bacillus sp. U andBacillus sp. W). In short-term contact incubation experiments with rich media, neither quartz crystal surface structures nor bacterial cell surfaces appeared to be changed. Likewise, significant biochemical dissolution and mechanical dislocation of SiO₂ (which would have indicated rapid bacterial weathering activities) could not be detected. The importance of quartz purity and crystalline structure for the initiation of weathering processes is discussed.     

Rubber particles from four different species, examined by transmission electron microscopy and electron-paramagnetic-resonance spin labeling, are found to consist of a homogeneous rubber core enclosed by a contiguous, monolayer biomembrane

The physical characteristics of rubber particles from the four rubber (cis-1,4-polyisoprene) producing species Euphorbia lactiflua Phil., Ficus elastica Roxb., Hevea brasiliensis Müll. Arg., and Parthenium argentatum Gray, were investigated using transmission electron microscopy (TEM) and electron-paramagnetic-resonance (EPR) spin labeling spectroscopy. Transmission electron microscopy showed the rubber particles to be composed of a spherical, homogeneous, core of rubber enclosed by a contiguous, electron-dense, single-track surface layer. The biochemical composition of the surface layer and its single-track TEM suggested that a monolayer biomembrane was the surface structure most compatible with the hydrophobic rubber core. The EPR spectra for a series of positional isomers of doxyl stearic acid, used to label the surface layer of the rubber particles, exhibited flexibility gradients and evidence for lipid-protein interactions for all four rubber particle types that is consistent with a biomembrane-like surface. The EPR spectra confirmed that the surface biomembrane is a monolayer. Thus, rubber particles appear similar to oil bodies in their basic architecture. The EPR spectra also provided information on protein location and degree of biomembrane penetration that correlated with the known properties of the rubber-particle-bound proteins. The monolayer biomembrane serves as an interface between the hydrophobic rubber interior and the aqueous cytosol and prevents aggregation of the particles. An unexpected observation for the probes in pure polyisoprene was evidence of an intrinsic flexibility gradient associated with the stearic acid molecule itself. Received: 22 May 1999 / Accepted: 21 June 1999     

Devescovinid features, a remarkable surface cytoskeleton, and epibiotic bacteria revisited in Mixotricha paradoxa, a parabasalid flagellate

Summary. This work reports on the flagellate systematics and phylogeny, cytoskeleton, prokaryote–eukaryote cell junction organisation, and epibiotic bacteria identification. It confirms the pioneer 1964 study on Mixotricha paradoxa and supplies new information. Mixotricha paradoxa has a cresta structure specific to devescovinid parabasalid flagellates, a slightly modified recurrent flagellum, and an axostylar tube containing two lamina-shaped parabasal fibres. However, many parabasal profiles are distributed throughout the cell body. There is a conspicuous cortical microfibrillar network whose strands are related to cell junction structures subjacent to epibiotic bacteria. The supposed actin composition of this network could not be demonstrated with anti-actin antibodies or phalloidin labelling. Four types of epibiotic bacteria were described. Bacillus-shaped bacteria with a Gram-negative organisation are nested in alternate rows on most of the surface of the protozoon. They induce a striated calyxlike junction structure beneath the adhesion zone linked to the cortical microfibrillar network. Slender spirochetes are attached by one differentiated end to the plasma membrane of the protozoon, forming knobs on the cell surface. Two very similar long rod-shaped bacteria are also attached on the knobs of the plasma membrane. A large spirochete attributed to the genus Canaleparolina is also attached to the protozoon. Observations on epibiotic bacteria and of their attachments are compared with several described epibiotic bacteria of symbiotic protozoa and with the results of the molecular identification of the epibiotic bacteria of M. paradoxa.Correspondence and reprints: Biologie des Protistes, Université Blaise Pascal de Clermont-Ferrand, Campus des Cézeaux, 63177 Aubiere Cedex, France.