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1.
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.  相似文献   

2.
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.  相似文献   

3.
The spermatozoa from testis and spermatheca of the plant-parasitic nematode Trichodorus similis Seinhorst, 1963 (Nematoda; Triplonchida; Trichodoridae) were studied with transmission electron microscopy (TEM), being the first study on spermatogenesis of a representative of the order Triplonchida and important to unravel nematode sperm evolution. Comprehensive results could only be obtained using high-pressure freezing (HPF) and freeze-substitution instead of chemical fixation, demonstrating the importance of cryo-fixation for nematode ultrastructural research. The spermatozoa from the testis (immature spermatozoa) are unpolarized cells covered by numerous filopodia. They contain a centrally-located nucleus without a nuclear envelope, surrounded by mitochondria. Specific fibrous bodies (FB) as long parallel bundles of filaments occupy the peripheral cytoplasm. No structures resembling membranous organelles (MO), as found in the sperm of many other nematodes, were observed in immature spermatozoa of T. similis. The spermatozoa from the uterus (mature or activated spermatozoa) are bipolar cells with an anterior pseudopod and posterior main cell body (MCB), which include a nucleus, mitochondria and MO appearing as large vesicles with finger-like invaginations of the outer cell membrane, or as large vesicles connected to the inner cell membrane. The peripheral MO open to the exterior via pores. In the mature sperm, neither FBs nor filopodia were observed. An important feature of T. similis spermatozoa is the late formation of MO; they first appear in mature spermatozoa. This pattern of MO formation is known for several other orders of the nematode class Enoplea: Enoplida, Mermithida, Dioctophymatida, Trichinellida but has never been observed in the class Chromadorea.  相似文献   

4.
Ca(2+) is one of the most universal and versatile signalling molecules and is involved in almost every aspect of cellular processes. Accumulating evidence suggests that Ca(2+) serves as a messenger in many growth and developmental processes and in plant responses to biotic and abiotic stresses. Numerous signals have been shown to induce transient elevation of cytoplasmic [Ca(2+)](cyt) in plants. The calcium free ions were detected cytochemically in Solanum tuberosum cv. Rywal tissues as a hypersensitive response (HR) from 10h to 5 days after a Potato virus Y (PVY) infection. Calcium was detected in vivo by its reaction with Alizarin S Red, producing an intense red staining in contact with calcium free ions. Calcium was found in the necrotic area of the epidermal and mesophyll cells 3 days after the PVY infection (when morphological symptoms on potato leaves appeared). Calcium ions were detected cytochemically in HR also by its reaction with potassium pyroantimonate. Inoculation with PVY(NTN) and also PVY(N) Wi induced a rapid hypersensitive response during which highly localised increased accumulation of electron-dense deposits of calcium pyroantimonate were detected. Calcium deposition was observed in necrotic and non-necrotic areas, starting from 10h after PVY infection. The deposits were present along ER cisternae, chloroplasts and mitochondria envelopes connected with PVY particles. The precipitates of calcium antimonate were detected near the nuclear envelope, inside karyolymph and along tracheary elements, especially when virus particles were present inside.  相似文献   

5.
The nematode spermatozoa represent a highly modified (aberrant) type of male gametes that lack a flagellum but for which the process of spermatogenesis culminates in the production of a crawling spermatozoon on the basis of the cytoskeletal component known as “major sperm protein”, or MSP. MSP is also known as an important hormone triggering oocyte maturation and ovulation in the model nematode Caenorhabditis elegans, where this protein was first identified. However, direct evidence of MSP localization and of its fate in nematode spermatogenic cells is rare. In this study, the spermatogenesis and sperm structure in the rhabditid nematode Acrobeles complexus (Rhabditida: Tylenchina: Cephalobomorpha: Cephaloboidea: Cephalobidae) has been examined with electron microscopy. Morphological observations were followed by high-pressure freezing and freeze-substitution fixation which allows post-embedding immunogold localization of MSP in all stages of sperm development using antibodies raised for MSP of C. elegans. In spermatocytes, synthetic activity results in the development of specific cellular components, fibrous bodies (FB) and membranous organelles (MO), which appear as FB-MO complexes where the filamentous matter of FB has been MSP-labeled. The spermatids subdivide into a residual body with superfluous cytoplasm, and a main cell body which contains nucleus, mitochondria and FB-MO complexes. These complexes dissociate into individual components, MO and FB, with the MSP being localized in FB. Immature spermatozoa from testes are opaque cells where a centrally located nucleus is surrounded by mitochondria, MO and FB clustered together, the MSP still being localized only in FB. Cytoplasm of mature spermatozoa from spermatheca is segregated into external pseudopods lacking organelles and a central cluster of mitochondria with intact MO surrounding the central nucleus. The FB ultimately disappear, and the MSP labeling becomes concentrated in the filamentous content of pseudopods and cytoplasm of the main cell body. Although the spermatogenesis and sperm structure of A. complexus is similar to that of many other rhabditid nematodes, their intact MO makes the morphology of the mature spermatozoa distinct from the “rhabditid pattern” and may be considered as a synapomorphy. The MSP localization in spermatogenic cells of A. complexus also follows the “rhabditid pattern” described in C. elegans and Ascaris spp. Our results and techniques of MSP labeling of A. complexus spermatogeneous cells reveal new possibilities to elucidate different research questions on MSP localization in nematodes related to C. elegans. Furthermore, the laboratory-cultured A. complexus strains can be used as a new and fascinating model to study MO and MSP functions in nematode reproduction.  相似文献   

6.
The ultrastructure of cells within shoot tips of S. tuberosum 'Désirée' was studied after different steps of the DMSO droplet cryopreservation method. After 2 h of DMSO treatment, cells contained numerous small vesicles, while at the same time mitochondria and chloroplasts had increased in size and vacuoles had assumed an irregular shape. After rapid cooling in liquid nitrogen, subsequent rewarming, and 1 h incubation there were no apparent changes in the ultrastructural organization of the cells, suggesting that they might be still intact. However, two days after rewarming, the meristematic dome area and part of the epidermis showed signs of extensive damage. Rupture of plasmalemma, plasmolysis and destruction of cell organelles as well as strong heterochromatisation of nuclei were observed. Survival and regeneration of cells were found mainly in leaf primordial regions. Here cells were very active, containing many mitochondria and intact or regenerating chloroplasts. Alternating temperature preculture of donor plants before shoot tip isolation improved the cryopreservation results (plant regeneration 46.5 percent) as compared to constantly warm precultured shoot tips (plant regeneration 20.0 percent), which showed slightly stronger damage after rewarming from liquid nitrogen.  相似文献   

7.
In this paper, to understand the roles of amorphous structures which were observed within the viromatrix of Rana grylio virus (RGV), an improved immunoelectron microscopy (IEM) method was developed to detect the localization of RGV in carp Epithelipma papulosum cyprinid (EPC) cells. Infected EPC cells were fixed with 4% paraformaldehyde-0.25% glutaraldehyde mixture, dehydrated completely, and embedded in LR White resin. This method allowed good ultrastructural preservation and specific labeling with anti-RGV antibodies. The results of IEM showed that colloidal gold mainly bound to the capsids of viral particles at the stage of viral assembly, while during the viral maturation colloidal gold bound to the envelop of virions. In addition, within the viromatrix, the amorphous structures, including dense floccules, membranous materials and tubules, also had strong colloidal gold signals, revealing that those amorphous structures were participated in RGV assembly. In contrast, no significant gold labeling signals were obtained in negative controls. The present study not only provided further evidence that amorphous structures within the viromatrix were involved in the process of RGV assembly, but also developed an improved IEM method for studying the interaction between iridovirus and host cells.  相似文献   

8.
Leaf salt glands of Limoniastrum guyonianum were examined by scanning and transmission electron microscopes and energy dispersive X-ray analysis (EDAX) system, after growing for three months on sandy soil with or without 300 mM NaCl. Results showed that salt glands were irregularly scattered on both leaf sides and sunk under the epidermal level. Salt excretion occurred in both conditions and is mainly composed of calcium and magnesium in control plants, and essentially sodium and chloride in plants subjected to salt treatment. A salt gland is comprised of collecting, accumulating, and central compartments, and is made up of total thirty-two cells. The collecting cells were characterized by large central vacuoles. Accumulating cells contain numerous, large, and unshaped vacuoles and rudimentary chloroplasts. The central compartment was comprised of four basal cells and each one is surmounted by an apical cell. The basal cells are granulated, containing large nucleus, numerous mitochondria, endoplasmic reticulum, ribosomes, polyribosomes, and small vacuoles or vesicles. Equally, the apical cells are rich in organelles. Application of 300 mM NaCl to the culture medium increased vacuoles number and size, and organelles density especially the mitochondria which suggests energy requirement for ions transport. The reduction in size and number of vacuoles toward the interior of salt glands of treated plants and the fusion of the smallest ones with the plasma membrane substantiate the implication of such vacuoles in salt excretion process. The current study which is the first report on L. guyonianum salt gland has provided an in-depth understanding on structure–function relationship in the multicellular salt glands.  相似文献   

9.
ATP is synthesized by F-type proton-translocating ATPases (F-ATPases) coupled with an electrochemical proton gradient established by an electron transfer chain. This mechanism is ubiquitously found in mitochondria, chloroplasts and bacteria. Vacuolar-type ATPases (V-ATPases) are found in endomembrane organelles, including lysosomes, endosomes, synaptic vesicles, etc., of animal and plant cells. These two physiologically different proton pumps exhibit similarities in subunit assembly, catalysis and the coupling mechanism from chemistry to proton transport through subunit rotation. We mostly discuss our own studies on the two proton pumps over the last three decades, including ones on purification, kinetic analysis, rotational catalysis and the diverse roles of acidic luminal organelles. The diversity of organellar proton pumps and their stochastic fluctuation are the important concepts derived recently from our studies.  相似文献   

10.
The present study evaluates by ultrastructural and immunohistochemical methods, the possible changes on muscular tissue affected by LLLI during a treatment, for example, in cases of temporomandibular joint disorders. Sixty male Wistar rats divided into 6 groups (n=10) received ten laser irradiations, with different energy densities (groups I-0; II-0.5; III-1.0; IV-2.5; V-5.0; and VI-20 J/cm(2)). Muscles were removed and processed for transmission electron microscopic and immunohistochemical (VEGF and VEGFR-2) analyses. Captured photomicrographs of immunohistochemistry and transmission electron microscopy were evaluated. It was observed in the irradiated muscles, mitochondria of different shapes and sizes, with increased plasticity evidenced by organelles in fusion, division and the presence of elongated structures with characteristics of mitochondria, proximity with the dilated sarcoplasmatic reticulum, suggesting organelles with large amounts of energy, and the presence of cytoplasmic protrusions in the capillaries with high dosages. All studied groups showed immunostainings for both markers (VEGF and VEGFR-2), but in general those who received higher doses also showed the markings more pronounced, suggesting dose-dependent biomodulation. It was concluded that the LLLI was able to modify the ultrastructural characteristics and immunohistochemical pattern of VEGF and VEGFR-2 in the masseter muscle of rats.  相似文献   

11.
Cytoplasm streaming is a fundamental process for the transport of molecules and organelles in plant cells. In vegetative filaments of the coenocytic green alga, Halimeda cuneata Hering, the spatial organisation of microtubules in the cytoplasmic layer, was observed under transmission electron microscopy. A cross section of a cortical filament shows a tubular cell wall enclosing a peripheral layer of cytoplasm with numerous chloroplasts, amyloplasts, nuclei, mitochondria and microtubules surrounding a small central vacuole. Towards the thallus medulla the central vacuole enlarges considerably and the cytoplasm becomes gradually reduced to a thin parietal layer, the number of organelles is reduced and the quantity of microtubules increases. Therefore, most of the thallus volume is occupied by the huge central vacuole which extends throughout the coenocytic filaments. Microtubules run longitudinally, being about 0.05 μm from each other. Some microtubules were observed in close association to cell organelles.  相似文献   

12.
The reactive oxygen species hydrogen peroxide (H2O2) was detected cytochemically in Solanum tuberosum cv. Rywal tissues as a hypersensitive response (HR) 24 and 48 h after a Potato virus Y (PVY) infection.Hydrogen peroxide was detected in vivo by its reaction with 3.3-diaminobenzidine, producing a reddish-brown staining in contact with H2O2. Hydrogen peroxide was detected in the necrotic area of the epidermal and mesophyll cells 24 and 48 h after the PVY infection. Highly localised accumulations of H2O2 were found within xylem tracheary elements, and this was much more intensive than in non-infected leaves. Hydrogen peroxide was detected cytochemically in HR also by its reaction with cerium chloride, producing electron-dense deposits of cerium perhydroxides.Inoculation with PVYNTN and also PVYN Wi induced a rapid hypersensitive response during which highly localised accumulations of H2O2 was detected in plant cell walls. The most intensive accumulation was present in the bordering cell walls of necrotic mesophyll cells and the adjacent non-necrotic mesophyll cells. Intensive electron-dense deposits of cerium perhydroxide were found along ER cistrenae and chloroplast envelopes connected with PVY particles. The precipitates of hydrogen peroxide were detected in the nuclear envelope and along tracheary elements, especially when virus particles were present inside. The intensive accumulation of H2O2 at the early stages of potato–PVY interaction is consistent with its role as an antimicrobial agent and for this reason it has been regarded as a signalling molecule.  相似文献   

13.
Diffusion tensor imaging (DTI) was performed on 25 patients with neurocysticercosis (NCC). The aim of this study was to investigate the changes in DTI measures during the evolutionary course of NCC lesions from vesicular to calcified stage in the brain. DTI measures were quantified from the NCC lesions of all patients. On the basis of conventional imaging findings, NCC lesions were classified into vesicular, vesicular colloidal, granular nodular and calcified stages. Significant inverse correlation was observed between the evolutionary stage of NCC lesion and mean diffusivity (MD; r=−0.748, P<0.001) and spherical anisotropy (CS; r=−0.585, P<.001) values. Significant direct correlations were observed between evolutionary stages of NCC lesion and mean fractional anisotropy (FA; r=0.575, P<0.001), linear anisotropy (CL; r=0.478, p<0.001) and planar anisotropy (CP; r=0.561, p<0.001) values. Successive decrease in MD values calculated from NCC lesions was observed, moving from vesicular to granular nodular stage. On FA, CL and CP maps, a significant increase in signal intensity value was observed in calcified as compared to other stages. We conclude that DTI measures may indicate the evolutionary changes in NCC from vesicular to calcified stage.  相似文献   

14.
The Atlantic jackknife clam, Ensis directus, is currently being researched as a potential species for aquaculture operations in Maine. The goal of this study was to describe the hemocytes of this species for the first time and provide a morphological classification scheme. We viewed hemocytes under light microscopy (using Hemacolor, neutral red, and Pappenheim’s stains) as well as transmission electron microscopy (TEM). The 2 main types of hemocytes found were granulocytes and hyalinocytes (agranular cells). The granulocytes were subdivided into large and small granulocytes while the hyalinocytes were subdivided into large and small hyalinocytes. The large hemocytes had both a larger diameter and smaller nucleus to cell diameter ratio than their smaller counterparts. A rare cell type, the vesicular cell, was also observed and it possessed many vesicles but few or no granules. Using TEM, granulocytes were found to contain both electron-lucent and electron-dense granules of various sizes. These numerous granules were the only structures that took up the neutral red stain. Hyalinocytes had few of these granules relative to granulocytes. Large hyalinocytes had both various organelles and large vesicles in their abundant cytoplasm while small hyalinocytes had little room for organelles in their scant cytoplasm. Total hemocyte counts averaged 1.96 × 106 cells mL−1 while differential hemocyte counts averaged 11% for small hyalinocytes, 12% for large hyalinocytes, 59% for small granulocytes, and 18% for large granulocytes. The results of this study provide a starting point for future studies on E. directus immune function.  相似文献   

15.
This paper investigates analytically the molecular-motor-assisted transport between the cell nucleus and cell membrane in an elongated cell, which allows the formulation of governing equations in a cylindrical coordinate system. This problem is relevant to biomimetic transport systems as well as to many biological processes occurring in living cells, such as the viral infection of a cell. The obtained analytical solution is shown to agree well with a high-accuracy numerical solution of the same problem. The developed analytical technique extends the applicability of the generalized Fourier series method to a new type of problems involving intracellular transport of organelles.   相似文献   

16.
Cratylia mollis is a native forage from the semi-arid region of Northeast, State of Pernambuco, Brazil, whose seeds have been considered an important lectin source. Multiple molecular forms of lectins, carbohydrate-binding proteins, have been purified from C. mollis seeds (Cra Iso) allowing several applications of these purified proteins. In this work seeds were processed for ultrastructural analysis and immunocytochemical localization of the two most abundant isolectins, Cra Iso 1 and Cra Iso 3, with glucose/mannose and galactose specificities, respectively. The ultrastructural analysis revealed a typical plant cell: organelles, nucleus and cellular wall were visualized. The localization of isolectins occurred mainly in the amorphous matrix of protein bodies, and in the cellular walls of the embryonic axis. The results showed that the isolectins, which differ in relation to carbohydrate specificity and glycosylation are located in the same cellular compartment suggesting different functions inside the same subcellular organelle. Cra Iso 1 and Cra Iso 3 distribution in the C. mollis seeds was consistent with the subcellular localization of several legume lectins.  相似文献   

17.
Supercooling sperm in liquid nitrogen vapour is a feasible and economic technique for the practical production. The study aimed to reveal the negative effects of this rapid freezing and thawing processes on Taihang black goat spermatozoa and to find out the changing of spermatozoa motility and ultrastructure by using CASA and TEM. Qualified semen samples, which collected from twenty Chinese Taihang black goats using artificial vagina were pooled and investigated the kinematics parameters and ultrastructural morphology. The results showed that freezing–thawing caused a significant reduction in the spermatozoon total motility (P < 0.001), in rapid and medium cell numbers (P < 0.001) and motility parameters (VAP, VSL, VCL, ALH and BCF) (P < 0.01). Immotile spermatozoa number was increased significantly after freezing–thawing (P < 0.001). In the ultrastructural analysis, the shape with a sperm nucleus characterized by ruptures, bend and deformity was observed. The plasma membranes were broken, and nucleoplasm erupted. The mitochondria in the middle piece were disturbed by partial absence or additional accumulations. Swelling, coiling, vacuolization and structural disorganization of mitochondria were also observed. In conclusion, Freezing–thawing procedure has a detrimental effect on motility, membrane integrity and mitochondria of goat spermatozoa. Transmission electron microscopy provides an intuitive observation to investigate deformity spermatozoa.  相似文献   

18.
The present ultramorphological, histological and ultrastructural study on the fat body of semi-engorged females of Amblyomma cajennense revealed that this tissue is diffuse and consists of strands of cells surrounding the tracheal trunks. Morphometric analysis showed that the cellular and nuclear areas of round-shaped trophocytes are larger than those of cuboidal trophocytes, indicating that the arrangement of the former provides more contact area with the haemolymph. In this species, the fat body is found right underneath the integument and around organs. It consists of two cell types that despite distinct morphological characteristics and locations in the tissue, present the same histological features. In this study, these cells were termed cuboidal trophocytes when arranged as strands of cells and present in larger numbers, and round-shaped trophocytes when lying on these strands and observed in fewer numbers. Histological observations revealed that both types of trophocytes have one nucleus in their cytoplasm and also exhibit numerous vacuoles of different sizes and contents. Ultrastructural examination revealed that the organelles more frequently observed were the vesicular and lamellar rough endoplasmic reticulum, and mitochondria with tubular crests, indicating that they might be involved in lipid synthesis. Smooth endoplasmic reticulum was not observed. Cuboidal trophocytes arranged in strands, despite being closely associated, do not exhibit fused plasma membranes. Rather, the fusion of basal lamina of two neighboring cells is occasionally observed, acting as a selective permeability barrier. Here, a new terminology for tick fat body is proposed. It is based on fat body location (parietal, when located right underneath the integument instead of peripheral; and perivisceral, when located around organs instead of central) terminologies previously suggest by Obenchain and Oliver and for the cells constituting them, cuboidal trophocytes when arranged as strands, and round-shaped trophocytes when lying on these strands. Nephrocytes were not observed in semi-engorged females of A. cajennense.  相似文献   

19.
Recent work using the concept of alpha-clustering in studies of nuclear structure and nuclear reactions is reviewed. The high symmetry and binding energy of the alpha-particle makes it likely that nucleons inside the nucleus can condense into alpha particles and live long enough to affect many properties of nuclei and also the cross-sections of nuclear reactions, particularly those with alpha-particles as the projectile or ejectile. The alpha-particles inside the nucleus may escape, and the resulting alpha decay is now quite well understood. An incident projectile may collide with a transient alpha-particle in the nucleus and knock it out, thus providing information on the degree of clustering. This model enables the direct part of the cross-sections of (n, α) reactions to be calculated. The alpha-particle mean field unifies many of the characteristics of alpha-particle structure and alpha-particle scattering. Many properties of light nuclei may be simply explained using the concept of alpha-clustering.  相似文献   

20.
We considered applicability of acoustic imaging technology for the detection of magnetic microparticles and nanoparticles inside soft biological tissues. Such particles are widely used for magnetically targeted drug delivery and magnetic hyperthermia. We developed a new method of ultrasonic synchronous tissue Doppler imaging with magnetic modulation for in vitro and in vivo detection and visualization of magnetic ultradisperse objects in soft tissues. Prototype hardware with appropriate software was produced and the method was successfully tested on magnetic microparticles injected into an excised pig liver.  相似文献   

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