Ultrastructural insights into tomato infections caused by three different pathotypes of Pepino mosaic virus and immunolocalization of viral coat proteins

This paper presents studies on an ultrastructural analysis of plant tissue infected with different pathotypes of Pepino mosaic virus (PepMV) and the immunolocalization of viral coat proteins. Because the PepMV virus replicates with a high mutation rate and exhibits significant genetic diversity, therefore, isolates of PepMV display a wide range of symptoms on infected plants. In this work, tomato plants of the Beta Lux cultivar were inoculated mechanically with three pathotypes representing the Chilean 2 (CH2) genotype: mild (PepMV-P22), necrotic (PepMV-P19) and yellowing (PepMV-P5-IY). The presence of viral particles in all infected plants in the different compartments of various cell types (i.e. spongy and palisade mesophyll, sieve elements and xylem vessels) was revealed via ultrastructural analyses. For the first time, it was possible to demonstrate the presence of crystalline inclusions, composed of virus-like particles. In the later stage of PepMV infection (14 dpi) various pathotype-dependent changes in the structure of the individual organelles (i.e. mitochondria, chloroplasts) were found. The strongest immunogold labeling of the viral coat proteins was also observed in plants infected by necrotic isolates. A large number of viral coat proteins were marked in the plant conductive elements, both xylem and phloem.     

Cytochemical localization of calcium and X-ray microanalysis of Catharanthus roseus L. infected with phytoplasmas

The potassium pyroantimonate (KPA) Ca²⁺ precipitation technique, X-ray microanalysis and Electron Energy Loss Spectroscopy carried out by transmission electron microscopy were used to analyze the Ca²⁺ distribution in Catharanthus roseus L. leaves infected with phytoplasmas belonging to different taxonomic groups, and in phytoplasma cells. The analysis revealed that the distribution of Ca²⁺ was different in healthy and diseased plants (where the KPA deposits were numerous) and no differences were observed in the tissues of the three types of infected C. roseus L. Since no KPA precipitates were visible in the phloem and on phytoplasma cells, it is likely that Ca²⁺ ions are not directly involved in phytoplasma replication, but, in infected cells is a response to the pathogen indicative of a higher Ca²⁺ in the plasmalemma.     

Multidimensional solid-state NMR spectroscopy of plant cell walls

Plant biomass has become an important source of bio-renewable energy in modern society. The molecular structure of plant cell walls is difficult to characterize by most atomic-resolution techniques due to the insoluble and disordered nature of the cell wall. Solid-state NMR (SSNMR) spectroscopy is uniquely suited for studying native hydrated plant cell walls at the molecular level with chemical resolution. Significant progress has been made in the last five years to elucidate the molecular structures and interactions of cellulose and matrix polysaccharides in plant cell walls. These studies have focused on primary cell walls of growing plants in both the dicotyledonous and grass families, as represented by the model plants Arabidopsis thaliana, Brachypodium distachyon, and Zea mays. To date, these SSNMR results have shown that 1) cellulose, hemicellulose, and pectins form a single network in the primary cell wall; 2) in dicot cell walls, the protein expansin targets the hemicellulose-enriched region of the cellulose microfibril for its wall-loosening function; and 3) primary wall cellulose has polymorphic structures that are distinct from the microbial cellulose structures. This article summarizes these key findings, and points out future directions of investigation to advance our fundamental understanding of plant cell wall structure and function.     

Biotrophic interaction of Sporisorium scitamineum on a new host—Saccharum spontaneum

Sporisorium scitamineum is a biotrophic smut fungus harbored inside the smut gall on the top internodal region of Saccharum spontaneum, a wild relative of sugarcane (Saccharum officinarum). The interactions of spined conidia of S. scitamineum with S. spontaneum were examined during the different stages of plant growth starting from the bud stage to the decaying stage. The spores in the soil from the polyetic inocula grew into confined epidermal cells of the buds and finally sporulated in the topmost internodal region. Hyphae invasion of the plant tissues were restricted to the point of infection. Culms of infected plants in late October sporulated, notably; hyphal sporulation produced shorter hyphal stolons. Remarkably, the nodal regions of infected plants had no spores and fragmented hyphae. On the basis of microscopic analyses, hyphae and spores were absent in all internodes above the ground till the topmost smut gall region. This result indicated that, S. scitamineum undergoes tissue-confined invasion of S. spontaneum. By associating culture medium method with polymerase chain reaction (PCR) on plant portions void of smut gall, S. scitamineum was not detected, indicating that colonization was not systemic. It was observed that the biotrophic interaction resulted in structural reorganization in the restricted region of infection forming erect cylindrical structure, in which the fungus was sandwiched between the central stalk and sheath, and possibly played a key role in preventing inflorescence. Comparatively, a significant difference in the rate of teliospores germination between reference Ustilago esculenta (26.6%, P < 0.05) and S. scitamineum (62.9%, P < 0.05) at 20° C was observed. This study also provides insights on the effect of different temperature regimes on the germination of S. scitamineum teliospores in vitro.     

Morphological characterization of the mouthparts of the vector leafhopper Psammotettix striatus (L.) (Hemiptera: Cicadellidae)

The leafhopper Psammotettix striatus (L.) (Hemiptera: Cicadellidae), is one of the most significant economic pests of wheat in Western China. This insect vectors a phytoplasma that causes wheat blue dwarf (WBD), a severe disease limiting wheat production in the Western China. A microscopic analysis of the ultrastructure of the mouthparts of the adult was conducted using scanning electron microscopy and the putative functions of the mouthparts were determined. The piercing-sucking mouthparts of P. striatus are of the conventional type comprising a three-segmented labium with a deep groove in the anterior side, a stylet fascicle consisting of two mandibular and two maxillary stylets, and an uppermost small cone-shaped labrum. The mandibular stylets, located laterad of the maxillary stylets, have sculpture on their tips, which may function in tearing plant tissue, cutting channels into the plant tissues, and attaching the body to the host plant during molting. The maxillary stylets are interlocked to form two separate compartments, a larger food canal and a smaller salivary canal. Two dendritic canals are also found in each maxilla and one in each mandible. Four kinds of sensilla were found on the labium: s. trichodea I, s. trichodea II, s. basiconic I, s. basiconic II. These may be involved in host recognition and are likely chemo- or mechanosensory, or both.     

Occurrence and ultrastructure of Albugo candida on a new host, Arabis alpina in Saudi Arabia

A population of Arabis alpina (Brassicaceae) growing in Saudi Arabia was observed to be infected for the first time by the Oomycete, Albugo candida. Both conventional chemical fixation and high pressure freezing followed by freeze substitution (HPF/FS) were used to prepare zoosporangia, intercellular hyphae, haustoria, invading host cells and host–parasite interface of A. candida for study with both scanning and transmission electron microscopy. Both fixations gave good preservation of ultrastructural details and data from the two sample types were highly complementary. Scanning electron microscopic observation revealed that mature zoosporangia of A. candida are spherical or ellipsoidal in shape and characterized by a smooth surface and faint terminal secession scar at each end. Transmission electron microscopic observation indicated that coenocytic intercellular hyphae are located in intercellular spaces of host leaf tissue forming haustoria in host mesophyll cell. Each haustorium is connected to intercellular hyphae by a narrow, slender neck which enclosed by a collar as a response of host cell to infection. The cytoplasm of the haustorium contains different organelles characteristic of the Oomycetes. No obvious responses are observed in host cell organelles following infection which may be due to the presence of a compatibility between the host and the Oomycete. Modifications of the host plasma membrane around the haustorium are detected. Many tubular elements were found to be continuous with the extrahaustorial membrane. This appears to be the first report of the presence of these tubular elements in case of A. candida haustoria. These tubular elements may increase membrane surface area and consequently increase the efficacy of nutrients uptake by haustoria from host cell.     

Elemental distribution and Pb speciation in vegetable and cereal seeds during germination by micro X-ray fluorescence and X-ray absorption near-edge structure

The distribution of the chemical elements in plants is closely correlated with biological functions. Micro X-ray fluorescence (μXRF) and X-ray absorption near-edge structure (XANES) spectrometry can be used to determine uptake, translocation, and accumulation of the elements in plants. In this research, μXRF and XANES were applied in studying distribution of the elements in the vegetable and cereal seeds during germination and the mobilization. It was found that (a) the distribution of the elements in the seeds was element specific; (b) K and Ca were mainly located in the embryo. Fe and Zn, as well as Mn and Cu, concentrated near the radicle and new developed root. Most of Pb was located in the new developing root; (c) the organic-Pb complexes functioned as the main components of translocation of Pb in the seeds during germination, and the formation of Pb₅(PO₄)₃Cl and Pb₃(PO₄)₂ in seed and plant tissues was the mechanism of tolerance and detoxification to Pb in plants.     

Maximal spontaneous CP-violation and left-right symmetry

An interesting form of quark mass matrices that arises in left-right symmetric models of weak interactions is discussed. It leads to a maximal CP-phase in the left-handed charged-current quark mixing matrix and to predictions for the mixing matrix elements U_us and U_cb in agreement with experiment, leaving U_ub as a free parameter. A way in which the model may be realized in a technically natural manner is described.     

The S exchange interaction in Gd doped La3-x□xS4

In this letter we show that the ESR line widths of Gd³⁺ in La₂S₃ (an insulator) and La₃S₄ (a metal) are the same at 0° K, but that in the latter it has a linear temperature dependence. Because of the identical crystal structures of these compounds this result can unequivically be interpreted as arising from an exchange interaction between the localized moment and the conduction electrons. The system La_3-x□_xS₄ therefore is an excellent host for studying the effects of localized moment-conduction electron interactions.     

Determining the elemental composition of Calotropis procera using X‐ray Analytical Microscopy

Caloptropis procera (Oshar) is a desert plant that did not receive much attention from the science community. The objective of this study was to investigate the elemental composition of the different parts of the plant using an X‐ray analytical microscope, to identify the elements naturally present in the plant and in the future detect the presence of any contaminants absorbed by the plants from the surrounding environment. Stalks, leaves and flowers from three Oshar plants were qualitatively and quantitatively analyzed. Leaves were scanned to establish the elemental spatial distribution within individual leaves. Subsequently, parts of the plants were dried, crushed and pulverized, then analyzed to determine elemental concentrations. The major elements present throughout the plant were Cl, K and Ca with varied concentrations. Other elements (Mg, Si, P, Fe, Sr, Mn and Br) are present in the leaf with various low concentrations of <5%. Major, minor and trace elements present in the various plant parts were determined. The outcome of this study will be used as a pilot for further investigations pertaining to the utilization of the Oshar plant for environmental cleaning purposes. Copyright © 2012 John Wiley & Sons, Ltd.     

Cytopathological potato virus Y structures during Solanaceous plants infection

The ultrastructural analysis of tobacco, potato and pepper tissues during infection with necrotic strains and the ordinary Potato virus Y strain of revealed the presence of virus inclusions not only in the epidermis and mesophyll but also in the vascular tissues. For the first time cytoplasmic inclusions were documented in companion cells and phloem parenchyma as well as in xylem tracheary elements. The ultrastructural features studied in this work consisted of mostly laminated inclusions (in the traverse and longitudinal section), which were frequently connected with enlarged cisternae of endoplasmic reticulum (ER) located in the direct vicinity of the cell wall attached to virus particles opposite to plasmodesmata. It was noticed that ER participates in synthesis and condensation of the PVY inclusions. During compatible interaction of tobacco and potato plants with PVY, amorphous and nuclear inclusions were observed. Such forms were not found in pepper tissues and potato revealing the hypersensitivity reaction to the infection with PVY necrotic strains. It was stated that the forms of cytoplasmic inclusions cannot serve as a cytological criterion to distinguish the potato virus Y strains and do not depend on host resistance level. Only in compatible interaction in Solanaceous plants tissues cytoplasmic inclusions were observed from the moment the morphological symptoms appeared. In the reaction of hypersensitivity, the inclusions were found on the 24th day following the infection with the PVY necrotic strains, whereas the symptoms were observed 3 days after the PVY infection.     

A new paradigm of bacteria-gut interplay brought through the study of Shigella

Bacteria-gut epithelial interplay and the mucosal immune response are the most critical issues in determining the fate of bacterial infection and the severity of diseases. Shigella species (abbreviated here as Shigella), the causative agent of bacillary dysentery (shigellosis), are highly adapted human pathogens that are capable of invading and colonizing the intestinal epithelium, which results in severe inflammatory colitis. Shigella secrete a large and diverse number (more then 50) of effectors via the type III secretion system (TTSS) during infection, some of which are delivered into the surrounding bacterial space and some others into the host cell cytoplasm and nucleus. The delivered effectors mimic and usurp the host cellular functions, and modulate host cell signaling and immune response, thus playing pivotal roles in promoting bacterial infection and circumventing host defense systems. This article overviews the pathogenic characteristics of Shigella, and highlights current topics related to the bacterial infectious stratagem executed by the TTSS-secreted effectors. Though bacterial stratagems and the molecular mechanisms of infection vary greatly among pathogens, the current studies of Shigella provide a paradigm shift in bacterial pathogenesis.     

Trichobilharzia regenti (Schistosomatidae): 3D imaging techniques in characterization of larval migration through the CNS of vertebrates

Migration of parasitic worms through the host tissues, which may occasionally result in fatal damage to the internal organs, represents one of the major risks associated with helminthoses. In order to track the parasites, traditionally used 2D imaging techniques such as histology or squash preparation do not always provide sufficient data to describe worm location/behavior in the host. On the other hand, 3D imaging methods are widely used in cell biology, medical radiology, osteology or cancer research, but their use in parasitological research is currently occasional. Thus, we aimed at the evaluation of suitability of selected 3D methods to monitor migration of the neuropathogenic avian schistosome Trichobilharzia regenti in extracted spinal cord of experimental vertebrate hosts. All investigated methods, two of them based on tracking of fluorescently stained larvae with or without previous chemical clearing of tissue and one based on X-ray micro-CT, exhibit certain limits for in vivo observation. Nevertheless, our study shows that the tested methods as ultramicroscopy (used for the first time in parasitology) and micro-CT represent promising tool for precise analyzing of parasite larvae in the CNS. Synthesis of these 3D imaging techniques can provide more comprehensive look at the course of infection, host immune response and pathology caused by migrating parasites within entire tissue samples, which would not be possible with traditional approaches.     

The democratic nuclear interaction in relative S states

A simple relative l=0 interaction is obtained by setting all relative matrix elements of a given spin equal to each other. The nuclear matrix elements are nearly equal for this interaction and the standard l=0 interactions such as the Kallio-Koltveit interaction, although the relative matrix elements are quite different.     

Comment on the spin-other-orbit and spin-spin interactions in atomic structure calculations

We have recalculated the interaction matrix elements for the spin-other-orbit interaction ford ² configurations and the matrix elements for the spin-other-orbit and spin-spin interactions forp ⁿ configurations. We conclude that Jones expressions for the offdiagonal elements are in error, while those of Hoire; Yamanouchi and Hoire are correct.     

TOF-SIMS investigation of Streptomyces coelicolor, a mycelial bacterium

Streptomyces coelicolor is a mycelial microorganism that produces several secondary metabolites, including antibiotics. The physiology of the organism has largely been investigated in liquid cultures due to ease of monitoring different physiological parameters and more homogeneous culture conditions. However, solid cultures reflect the natural physiology of the microorganism better, given that in its natural state it grows in the soil. Imaging mass spectrometry with TOF-SIMS and C₆₀⁺ primary ion beams offers a potential route to studying chemical changes at the molecular level, both intracellular and extracellular that can help in understanding the natural physiology of the microorganism. Here, we report the application of the technique for studying the lateral distribution of the chemical species detected in a population, grown in both liquid and solid cultures. The capability of the technique for studying biological systems with minimal system intervention is demonstrated.     

Study of K−p→K1(890)n at 13 GeV

The results of a wire chamber spectrometer experiment studying $\text{K}$¹ (890) production in the reaction K^?p→ K^?π⁺n at 13 GeV are presented. Strong forward structure is observed for |t|< m²_π in the s-channel density matrix elements and differential cross section. These features are similar to those observed in π^?p→?⁰n data and are characteristics of π exchange. In contrast in the intermediate, |t| ～ 0.2 GeV², and large momentum transfer regions $\text{K}$¹ (890) production is demonstrated by the natural parity ??A₂ exchange contribution.     

Collective isoscalar monopole and quadrupole oscillations with l-pole—l-pole interactions: A sum rule approach

Appropriate sum rules are shown to reproduce the results of microscopic calculations made in the random phase approximation for the soluble Hartree model with l-pole—l-pole interactions. This approach has the merit to avoid the problem of studying ground state correlations.     

Some current challenges in space plasma physics

An overview of the subject of space plasma physics is attempted. This encompasses a discussion of various ground-based and satellite studies of the near-Earth space environment, with its marked temporal and spatial variations, theoretical investigations, and numerical simulations. The important distinction is drawn between passive studies of elements of the natural system, aimed at the difficult task of understanding the self-consistent evolution of some natural processes, and active investigations, which either probe or modify natural processes, in order to improve understanding of cause and effect (e.g. of energy transformations) within the system. Linear, nonlinear, and chaotic processes can occur. Attention is paid to:

1. Energetic events on the Sun,
2. Large-scale features of the interplanetary medium, both observed directly and via scintillations of distant radio stars,
3. The magnetosphere in general, and the aurora in particular,
4. Wave-particle interactions, especially cyclotron resonance phenomena, and
5. The ionosphere, particularly when heated by powerful radio waves.

The approach adopted is to place these special topics in the context of the Solar Terrestrial Energy Program (STEP) of ICSU's SCOSTEP. Finally, mention is made of some benefits to humanity of studies of space plasma physics phenomena.