Light and Scanning Electron Microscopy of Greenbug Aphid Damage in Wheat Using the Same Section

A method has been developed to enable correlative light microscopy (LM) and scanning electron microscopy (SEM) on the same section of wheat (Triticum aestivum L.) leaves infested by greenbug aphids (Schizaphis gra-minum Rondani). Segments of infested leaf tissue were fixed, embedded in paraffin, sectioned, and affixed to slides by standard histological techniques. Serial sections were viewed by LM as temporary mounts in xylene. Sections of interest were identified and re-embedded in fingernail polish, affixed to aluminum stubs, freed of polish with ethyl acetate or acetone, and sputter-coated for SEM. SEM of re-embedded leaf sections showed excellent preservation of leaf anatomy. The same aphid tracks and regions of cell damage identified by LM were visible. SEM increased resolution and provided a much clearer sense of the three-dimensional relations involved in the interaction between plant and insect.     

Light and Scanning Electron Microscopy of Greenbug Aphid Damage in Wheat Using the Same Section

A method has been developed to enable correlative light microscopy (LM) and scanning electron microscopy (SEM) on the same section of wheat (Triticum aestivum L.) leaves infested by greenbug aphids (Schizaphis gra-minum Rondani). Segments of infested leaf tissue were fixed, embedded in paraffin, sectioned, and affixed to slides by standard histological techniques. Serial sections were viewed by LM as temporary mounts in xylene. Sections of interest were identified and re-embedded in fingernail polish, affixed to aluminum stubs, freed of polish with ethyl acetate or acetone, and sputter-coated for SEM. SEM of re-embedded leaf sections showed excellent preservation of leaf anatomy. The same aphid tracks and regions of cell damage identified by LM were visible. SEM increased resolution and provided a much clearer sense of the three-dimensional relations involved in the interaction between plant and insect.     

Transmission Comparisons of Cucumber Mosaic Virus Subgroup I and II Isolates by Different Aphid Species

To investigate the transmission differences between Cucumber mosaic virus (CMV) subgroup isolates, we carried out a comparative study with five aphid species Myzus persicae, Aphis gossypii, Lipaphis erysimi, Aphis craccivora and Megoura viciae in laboratory and field experiments to evaluate spread of CMV Subgroup I NX and subgroup II AG isolates in tobacco. Both NX and AG varied in transmission efficiency by the five aphids, and our transmission results revealed important differences in transmission efficiency of two isolates by Myzus persicae and Aphis gossypii. In contrast, significant transmission differences were not detected with Lipaphis erysimi, Aphis craccivora or Megoura viciae. Interestingly, the overall transmission efficiencies of the two different subgroup strains were almost equal when field transmissions were tested with mixed populations of the five aphid species. Our results together with our previously reported experiments on competition of CMV subgroup isolates in tobacco suggest that variations in aphid vector populations contribute substantially to the epidemic potential of CMV subgroup isolates.     

Alpha-synuclein,especially the Parkinson's disease-associated mutants,forms pore-like annular and tubular protofibrils

Two mutations in the alpha-synuclein gene (A30P and A53T) have been linked to autosomal dominant early-onset Parkinson's disease (PD). Both mutations promote the formation of transient protofibrils (prefibrillar oligomers), suggesting that protofibrils are linked to cytotoxicity. In this work, the effect of these mutations on the structure of alpha-synuclein oligomers was investigated using electron microscopy and digital image processing. The PD-linked mutations (A30P and A53T) were observed to affect both the morphology and the size distribution of alpha-synuclein protofibrils (measured by analytical ultracentrifugation and scanning transmission electron microscopy). The A30P variant was observed to promote the formation of annular, pore-like protofibrils, whereas A53T promotes formation of annular and tubular protofibrillar structures. Wild-type alpha-synuclein also formed annular protofibrils, but only after extended incubation. The formation of pore-like oligomeric structures may explain the membrane permeabilization activity of alpha-synuclein protofibrils. These structures may contribute to the pathogenesis of PD.     

Microscopic analysis and seasonality of gemma production in the freshwater red alga Hildenbrandia angolensis (Hildenbrandiales, Rhodophyta)

The development and release of the unique vegetative propagules of the freshwater encrusting alga Hildenbrandia angolensis Welwitsch ex West et West, gemmae, were studied using several different microscopic and histochemical techniques. In addition, the seasonality of gemma production was monitored bimonthly over a 12‐month period in two spring‐fed streams in Texas, USA. Gemmae differentiate within the thallus and are subsequently released from the surface of the crust. Release of the gemmae most likely occurs by digestion of surrounding cells, as suggested by the presence of starch granules and lipid globules in the region between the released gemma and the thallus. The initial separation of the gemmae from the thallus occurs from the sides of the gemma or the bottom, or possibly simultaneously. Contrary to previous studies, we have observed that gemma production occurs endogenously within the thallus of freshwater Hildenbrandia, rather than on the surface of the crust in raised structures. Histochemical tests and electron microscopic examination indicate that the cells of the gemmae contain a large amount of floridean starch. The starch granules frequently form rings surrounding the nuclei of both gemma and thallus cells; a feature infrequently reported for florideophyte red algae. Our seasonality investigations indicate that large fluctuations in gemma production occur over 1 year, but at least some gemma production continues year‐round in the streams examined.     

Cytopathology of the soybean looper,Pseudoplusia includens,infected with the Pseudoplusia includens icosahedral virus

Ultrastructural responses of soybean looper cells of various tissues infected with Pseudoplusia includens icosahedral virus (PIIV), a newly characterized RNA virus [Y. C. Chao, H. A. Scott, and S. Y. Young (1983)J. Gen. Virol.64, 1835–1838], were studied in situ. Most cells of fat body and epidermis consistently contained virus particles and associated cytopathic structures. Virus particles, corresponding to those of purified PIIV in morphology and size, always occurred in the cytoplasm either in membrane-bound virogenic stroma and/or freely in the ground cytoplasm. Virogenic stroma, which appeared to be distinct from those induced by other insect viruses, consisted of electron-dense matrix material, in which virus particles were embedded, and membranous vesicles, 70 or 80 nm in diameter, containing nucleic acid-like fibrils. Virus particles in virogenic stroma occurred as hexagonally arranged crystalline arrays made up primarily of homogeneously dense particles, while those in the ground cytoplasm were dispersed randomly and had an electron-lucent central core. Extremely large numbers of virus particles were also located in noncellular cuticle layers of the integument.     

The Structure and Morphogenesis of the Ventral Ciliature in Paraurostyla hymenophora*

SYNOPSIS. The structure and morphogenesis of the ventral ciliature of Paraurostyla hymenophora (Stokes) are described. The oral primordium apparently originates in association with transverse cirrus #6, from which it migrates anteriorly simultaneous with kinetosomal proliferation. The primordium eventually forms an elongate ciliary field from which the future opisthe's fronto-ventro-transverse (FVT) and undulating membrane primordial fields arise. Concomitantly, the future proter's FVT primordial field is initiated by the disaggregation of frontal cirri #4, #5, and #6. Primordia then develop simultaneously within marginal and ventral cirral rows by a disaggregation of cirri within the respective rows, and do not give rise to new cirri until the FVT fields complete segregation into discrete cirri. Near the completion of cirral production from the FVT primordia, each ventral cirral primordium (VCP) forms the 2 rightmost transverse cirri. Segregation of new cirri within the marginal cirral primordia and VCP then occurs, eventually replacing all old cirri within their respective marginal and ventral cirral rows. At the end of cortical morphogenesis, all old ciliary organelles, with the exception of the adoral zone of membranelles, are either reorganized or replaced. These results suggest an evolutionary affinity between the ventral and marginal cirral rows and raise questions about the control of the developmental competence of individual primordia.     

Structure and development of stellate trichomes in Andradea ftoribunda Fr. Allem. (Nyctaginaceae)

LOURO, R. P., MIGUENS, F. C. & MACHADO, R. D., 1992. Structure and development of stellate trichomes in Andradea ftoribunda Fr. Allem. (Nyctaginaceae). Trichomes occur on both faces of young leaves. They are peltate-stellate on the abaxial face, and comprise a stalk and radiating cells with a rudimentary central apex. On the adaxial face the trichomes arc stellate with a large apex comprising one to three cells. In both cases the stalk is formed by three to six cells of which the most distal may contain a tannoid substance. In the adult leaf only the abaxial surface exhibits stellate trichomes, with two to three celled stalks. The central region of radial cells is depressed. On the adaxial side the hairs are shed during maturation of the leaf.     

Adhesion of Plasmodium gallinaceum Ookinetes to the Aedes aegypti Midgut: Sites of Parasite Attachment and Morphological Changes in the Ookinete

Plasmodium gallinaceum ookinetes adhered to Aedes aegypti midgut epithelia when purified ookinetes and isolated midguts were combined in vitro. Ookinetes preferentially bound to the microvillated luminal surface of the midgut, and they seemed to interact with three types of structures on the midgut surface. First, they adhered lo and migrated through a network-like matrix, which we have termed microvilli-associated network, that covers the surface of the microvilli. This network forms on the luminal midgut surface in response to blood or protein meals. Second, the ookinetes bound directly to the microvilli on the surface of the midgut and were occasionally found immersed in the thick microvillar layer. Third, the ookinetes associated with accumulations of vesicular structures found interspersed between the microvillated cells of the midgut. The origin of these vesicular structures is unknown, but they correlated with the surface of midgut cells invaded by ookinetes as observed by TEM. After binding to the midgut. ookinetes underwent extensive morphological changes: they frequently developed one or more annular constrictions, and their surface roughened considerably, suggesting that midgut components remain bound to the parasite surface. Our observations suggest that, in a natural infection, the ookinete interacts in a sequential manner with specific components of the midgut surface. Initial binding to the midgut surface may activate the ookinete and cause morphological changes in preparation for invasion of the midgut cells.     

Formulation and evaluation of ethosomes for transdermal delivery of lamivudine

The purpose of the present research was to investigate the mechanism for improved intercellular and intracellular drug delivery from ethosomes using visualization techniques and cell line study. Ethosomal formulations were prepared using lamivudine as model drug and characterized in vitro, ex vivo and in vivo. Transmission electron microscopy, scanning electron microscopy, and fluorescence microscopy were employed to determine the effect of ethosome on ultrastructure of skin. Cytotoxicity and cellular uptake of ethosome were determined using T-lymphoid cell line (MT-2). The optimized ethosomal formulation showed 25 times higher transdermal flux (68.4 +/- 3.5 microg/cm(2)/h) across the rat skin as compared with that of lamivudine solution (2.8 +/- 0.2 microg/cm(2)/h). Microscopic studies revealed that ethosomes influenced the ultrastructure of stratum corneum. Distinct regions with lamellar stacks derived from vesicles were observed in intercellular region of deeper skin layers. Results of cellular uptake study showed significantly higher intracellular uptake of ethosomes (85.7% +/- 4.5%) as compared with drug solution (24.9% +/- 1.9%). The results of the characterization studies indicate that lipid perturbation along with elasticity of ethosomes vesicles seems to be the main contributor for improved skin permeation.     

Morphology of the proboscis of Hubrechtella Juliae (nemertea,pilidiophora): Implications for pilidiophoran monophyly

The proboscis of Hubrechtella juliae was examined using transmission electron microscopy, scanning electron microscopy, and confocal laser scanning microscopy to reveal more features of basal pilidiophoran nemerteans for morphological and phylogenetic analysis. The proboscis glandular epithelium consists of sensory cells and four types of gland cells (granular, bacillary, mucoid, and pseudocnidae‐containing cells) that are not associated with any glandular systems; rod‐shaped pseudocnidae are 15–25 μm in length; the central cilium of the sensory cells is enclosed by two rings of microvilli. The nervous plexus lies in the basal part of glandular epithelium and includes 26–33 (11–12 in juvenile) irregularly anastomosing nerve trunks. The proboscis musculature includes four layers: endothelial circular, inner diagonal, longitudinal, and outer diagonal; inner and outer diagonal muscles consist of noncrossing fibers; in juvenile specimen, the proboscis longitudinal musculature is divided into 7–8 bands. The endothelium consists of apically situated support cells with rudimentary cilia and subapical myocytes. Unique features of Hubrechtella's proboscis include: acentric filaments of the pseudocnidae; absence of tonofilament‐containing support cells; two rings of microvilli around the central cilium of sensory cells; the occurrence of subendothelial diagonal muscles and the lack of an outer diagonal musculature (both states were known only in Baseodiscus species). The significance of these characters for nemertean taxonomy and phylogeny is discussed. The proboscis musculature in H. juliae and most heteronemerteans is bilaterally arranged, which can be considered a possible synapomorphy of Hubrechtellidae + Heteronemertea (= Pilidiophora). J. Morphol. 274:1397–1414, 2013. © 2013 Wiley Periodicals, Inc.     

Accumulation,localisation, and toxic effects of cadmium in the liverwort Lunularia cruciata

Summary. Accumulation, tissue and intracellular localisation, and toxic effects of cadmium were investigated in the liverwort Lunularia cruciata. The results of analyses carried out by atomic absorption spectrometry on single plants showed that the cadmium accumulation was dose- and time-dependent. Cadmium localisation was assessed by X-ray scanning electron microscopy microanalysis in gemmalings and in the different tissues of the thallus and by X-ray transmission electron microscopy microanalysis at the cellular level. The metal preferentially accumulated in the hyaline parenchyma and at the base of the gemma cups. Inside the cell, cadmium accumulated in the vacuoles and the cell wall. Metal accumulation was accompanied by a concomitant increase in sulphur content within the vacuoles of stressed cells. Gel-permeation chromatography showed that most of the cadmium was associated with a low-molecular-mass fraction eluting at a ratio of elution volume to void volume corresponding to that of phytochelatins. The excess of sulphur deposited in the vacuoles may well have been caused by the stress-induced synthesis of phytochelatins. At the ultrastructural level, sublethal concentrations of cadmium caused alterations of the fine structure of the cells, inducing marked alterations of the chloroplast structure. Cadmium also induced a dose-dependent inhibition of apical thallus growth and gemma germination.Correspondence and reprints: Department of Plant Biology, University Federico 11, via Foria 223, 80132 Naples, Italy.     

Ultrastructure and distribution of sensilla on the antennae of female fig wasp Eupristina sp. (Hymenoptera: Agaonidae)

In the species‐specific and obligate mutualism between the fig (Moraceae: Ficus spp.) and its pollinator (Hymenoptera: Agaonidae), the continuity of lifecycle of both partners completely depends on the female pollinator's ability to detect receptive figs. To better understand the chemical location mechanism, we examined the antennae and their sensilla of the female fig pollinator Eupristina sp. using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The antennae of female Eupristina sp. are geniculated, and in total, there were seven types of sensilla found on the antennae: two types of multiporous placoid sensilla (type 1 is sausage‐like and type 2 is rounded), sensilla trichodea (ST), basiconic sensilla (BS), chaetica sensilla (ChS), coeloconic sensilla (CoS), and one specialized sensillum classified as sensillum obscurum (SO). We described external morphology, abundance, distribution, ultrastructure and discussed putative functions. We inferred from their ultrastructures as chemoreceptors that two types of multiporous placoid sensilla, BS and CoS, were innervated by sensory neurons. The aporous type ST, ChS, and SO were not innervated by dendrites which may function as mechanoreceptor/proprioceptor. These results were also discussed in relation to the interaction between Eupristina sp. and its host fig.     

Comparative study of the corneal epithelium in some reptiles inhabiting different environments

El‐Bakry, A.M. 2011. Comparative study of the corneal epithelium in some reptiles inhabiting different environments. —Acta Zoologica (Stockholm) 92 : 54–61. The vertebrate cornea functions in either aquatic or aerial environments and in some cases in both. In terrestrial and aerial vertebrates, the cornea contributes most of the refractive powers of the eye because of the large variation in refractive index between the air and the cornea. The present study aimed to examine and compare the main features of the corneal epithelial surface of three reptilian species related to three different families (Caretta caretta, Varanus griseus and Mabuya quinquetaeniata) and inhabiting different environment, by light, scanning (SEM) and transmission electron microscopy. The mean epithelial cell densities of the species of the study were 8.670 ± 3.134, 5.945 ± 2.144 and 2.124 ± 713 respectively. The corneal epithelium of the three species observed by SEM showed a similarity to one another indicating that the apical cell surfaces possess regular polygonal cells with varieties of microprocesses. These microprocesses were represented by microplicae, numerous microvilli and some long microridges in C. caretta, microplicae and minute microholes in V. griseus and microplicae intermingled with short microvilli in M. quinquetaeniata. According to the densities of these microprocesses, three polymorphic cell types (light, medium and dark) appeared in C. caretta, light and medium cell types were observed in V. griseus and medium and dark cell types were noticed in M. quinquetaeniata. Different types of tight adhesions were observed by transmission electron microscopy between the cell borders of the epithelial cells which differ according to environment where the species occupy. In conclusion, variation in the structure of the corneal epithelial cells appears to be related to the living environment, such as aerial, terrestrial and aquatic ones, which is occupied by every species.     

Transmission of Zucchini yellow mosaic virus by Colonizing and Non-colonizing Aphids in Greece and New Aphid Species Vectors of the Virus

Nineteen aphid species were tested for their ability to transmit Zucchini yellow mosaic virus (ZYMV) from and to zucchini under laboratory conditions. Sixteen species were found to be new vectors of ZYMV (i.e. Aphis craccae, Aphis fabae, Aphis nerii, Aulacorthum solani, Brachycaudus cardui, Brevicoryne brassicae, Hyalopterus pruni complex, Hyperomyzus lactucae, Macrosiphoniella sanborni, Macrosiphum rosae, Metopolophium dirhodum, Myzus cerasi, Rhopalosiphum maidis, R. padi, Semiaphis dauci and Sipha maydis). Their transmission efficiency by a single aphid was low (0.1–4.2%). Myzus persicae was used as a control and was the most efficient vector (41.1%, one aphid per plant). Hayhurstia atriplicis, Myzus ascalonicus and Sitobion avenae did not transmit the virus. In four out of six new vectors assayed in arena tests for propensity estimation, propensity was higher than efficiency. Data from an experimental zucchini field in northern Greece revealed a high correlation between ZYMV spread and alatae of the vector species. The most abundant aphid vectors during 2 years experimentation were M. persicae, Aphis gossypii and Aphis spiraecola. The possible role of the 16 new and the previously known aphid vectors in the epidemiology of ZYMV was investigated using data of transmission efficiency combined with the captures of their alatae in the Greek net of a Rothamsted type suction trap.     

Antennal sensory organs and glands of the harlequin ladybird,Harmonia axyridis

Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) is a sub‐cosmopolitan species. Native to Asia, it has been released during the 20th century for classical and augmentative biological control of several herbivorous insects, mostly aphids and coccids. Despite its recognized positive impact on biological control, H. axyridis is now considered among the most dangerous invasive species in Europe and in most places where it has established. This is mostly due to its ability to reduce the populations of native predatory species of the same trophic guild. When exploring a new area, H. axyridis adults use semiochemical cues to acquire information about the habitat. Presumably, these cues are perceived by the sensilla located on the antennae. Surprisingly, in spite of the huge literature existing on H. axyridis, the antennal sensory organs have been poorly characterized. Here, we used scanning and transmission electron microscopy (SEM, TEM) techniques to study H. axyridis antennae, with focus on the various types of sensilla and their distribution in male and female individuals. The presence of various classes of antennal sensilla belonging to the main types described in insects (chemoreceptors, mechanoreceptors, and thermo‐hygroreceptors) was highlighted, as well as the widespread presence of antennal glands. The investigations showed some peculiar characteristics not known in Coccinellidae, such as the concentration of sensory structures at the level of the distal part of the apical antennomere and the discovery of antennal glands associated with it. No sexual dimorphism was revealed, neither for the general structure of the antenna (similar number of antennomeres and presence of modifications), nor for the total length and width of the antenna, the relative size of the antennomeres, the types of antennal sensilla, of their distribution and abundance. The potential relevance of these sensory structures and antennal glands, reported for the first time in Coccinellidae, is discussed in the context of intra‐ and interspecific communication.     

Operando Observations of SEI Film Evolution by Mass‐Sensitive Scanning Transmission Electron Microscopy

The solid electrolyte interphase (SEI) spontaneously formed on anode surfaces as a passivation layer plays a critical role in the lithium dissolution and deposition upon discharge/charge in lithium ion batteries and lithium‐metal batteries. The formation kinetics and failure of the SEI films are the key factors determining the safety, power capability, and cycle life of lithium ion and lithium‐metal batteries. Since SEI films evolve with the volumetric and interfacial changes of anodes, it is technically challenging in experimental study of SEI kinetics. Here operando observations are reported of SEI formation, growth, and failure at a high current density by utilizing a mass‐sensitive Cs‐corrected scanning transmission electron microscopy. The sub‐nano‐scale observations reveal a bilayer hybrid structure of SEI films and demonstrate the radical assisted SEI growth after the SEI thickness beyond the electron tunneling regime. The failure of SEI films is associated with rapid dissolution of inorganic layers when they directly contact with the electrolyte in broken SEI films. The initiation of cracks in SEI films is caused by heterogeneous volume changes of the electrodes during delithiation. These microscopic insights have important implications in understanding SEI kinetics and in developing high‐performance anodes with the formation of robust SEI films.     

Plant regeneration,origin, and development of shoot buds from root segments of <Emphasis Type="Italic">Melia azedarach</Emphasis> L. (<Emphasis Type="Italic">Meliaceae</Emphasis>) seedlings

Summary In vitro regeneration of plants from root culture of Melia azedarach seedlings was obtained. The origin and mode of development of the regenerated shoot buds were studied by means of histological analysis and scanning electron microscopy (SEM). Maximum shoot bud regeneration was achieved when root segments were cultured on Murashige and Skoog (MS) medium at quarter strength with 3% sucrose and 0.44 μM benzyladenine (BA) and kept under light (116 μmol m⁻² s⁻¹). Shoot bud elongation was achieved on MS with 0.44 μM BA, 0.46 μM kinetin (KIN), and 3.26 μM adenine sulphate (AD). Regenerated shoots were rooted on MS with 12.26 μM indole-3-butyric acid (IBA) for 4 d and subsequently in MS lacking plant growth regulators for 26 d. Plants were established in a potting substrate. Histological analysis of roots from intact seedlings (without treatment) demonstrated that during the early life of the roots, M. azedarach lacks preformed buds. In contrast, when the roots were excised and cultured in vitro, the histology and SEM observations revealed that buds originated from meristematic groups of cells, which had been formed from the pericycle and several layers beneath. These meristematic groups of cells grew towards the periphery of the cortex by crushing the outer layer of cortical cells. Further develoment led to the differentiation of leaf primordia and a shoot apical meristem.     

纤毛虫念珠异列虫射出胞器的超微结构观察

应用扫描电镜术和透射电镜术显示,纤毛虫念珠异列虫(Anteholosticha monilata)的射出胞器早期发生在细胞质深处,附近有不同类型的囊泡结构。成熟后射出胞器向表膜迁移,结构由不同电子密度片层的体部、结晶状的中心轴杆部和多层膜的帽部组成。受外界刺激时胞器冲破皮层射出,形态呈"蘑菇"状。据上述观察结果推测:该射出胞器具有防御作用,它可能起源于高尔基体活动产生的小泡;在亲缘关系较近的纤毛虫中,其射出胞器可能具有相似的分化特征。