Ultrastructure of meristematic epidermal cells of maize root under water deficit conditions

Summary Structural components of meristematicZea mays primary root epidermal cells were observed after osmotic stress of the nutrient medium (12.5 atm.) and after rehydration. After non-lethal osmotic stress (24 hours), aggregation of nuclear chromatin, parallel ER arrangement, and reduction of mitochondrial cristae were found. Increased number of Golgi cisternae indicated vesicle production inhibition, and microtubules were absent in treated cells, although plastid structure remained unchanged. After lethal osmotic stress (48 hours), fragmentation of cytoplasmic membranes and a more severe structure damage of all cellular components occurred. Structure of the nucleus, mitochondria, Golgi apparatus and microtubules reappeared in the cells after rehydration. The only new feature of these cells was occurrence of smooth ER, which may indicate that the ER system has acquired a different function in regenerated cells.     

Chilling root temperature causes rapid ultrastructural changes in cortical cells of cucumber (Cucumis sativus L.) root tips

Examination of root tips from cucumber (Cucumis sativus L.) seedlings grown at 8 degrees C for varying periods ranging from 15 min to 96 h, showed marked changes in the ultrastructure of cortical cells within only 15 min of exposure. Greater parts of the cortex were affected with longer periods of exposure, but the sequence of morphological changes in cell components was similar to that found for the roots exposed for 15 min. The effect of chilling injury included alterations in cell walls, nuclei, ER, mitochondria, plastids, and ribosomes. The extent of alterations varied greatly among cells, moderate to severe alterations to cell components being observable among adjoining cells. The measurements of root pressure using the root pressure probe showed a sudden, steep drop in the root pressure in response to lowering of the temperature of the bathing solution from 25 degrees C to 8 degrees C. These observations are discussed in the light of the information available on the ultrastructural and biochemical characteristics of the effect of cold exposure in chilling-sensitive plants.     

Drought-induced Short Roots in Arabidopsis thaliana: Structural Characteristics

In Arabidopsis thaliana, as in other Brassicaceae species, a progressive drought stress induced changes in root morphogenesis: from a threshold plant water deficit, the new emerging roots remain short, hairless and often take a tuberized shape at their base while drought persists. The organization of these drought-induced roots was examined in light microscopy in Arabidopsis thaliana, Columbia wild-type ecotype, and compared to the normal, well-watered lateral roots. The main structural traits were the absence of elongation zone, the arrest of cell cap expansion, the lack of root hairs (despite epidermal differentiation in trichoblasts and atrichoblasts) and the radial enlargement of epidermal and cortical cells. The early differentiation, close to the short root apex, of large and highly lignified metaxylem elements, the absence of starch accumulation in hypertrophied cortical cells appeared to be characteristic of the species Arabidopsis, as compared to other Brassicaceae. These structural alterations are discussed in terms of drought-induced changes in gene expression with regard to similar modifications described in root morphogenesis and root hair-defective Arabidopsis mutants.     

Ultrastructure of differentiating epidermal cells of maize root under water deficit conditions

The ultrastructure of differentiating epidermal cells of maize root (within the distance of 1 and 2 mm from the root body apex) were studied under conditions of non-lethal and lethal osmotic stresses of nutrient medium containing polyethylene glycol 4000, as well as regeneration of their ultrastructure following rehydration. The structural response to water deficit of the cells investigated was dependent on both the stress duration and the stage of their ontogenic development. Following non-lethal stress, in younger cells investigated (1 mm), condensation of nuclear chromatin, decrease of polyribosomes and increased density of free ribosomes in cytoplasm, reduction of mitochondrial cristae and occurrence of intramitochondrial inclusions, less compact dictyosomes were observed. Pastid structure remained unchanged. Microtubules were lacking in treated cells. In the more differentiated cells (2 mm) protoplast retreat from cell walls was also observed as well as a general decrease of ribosomes and ER elements in parietal cytoplasm, an increased number of intramitochondrial inclusions and mitochondrial membrane fragmentation. In these cells, Golgi apparatus was also lacking. The ultrastructure regeneration of the more differentiated cells was less pronounced. Lethal osmotic stress would cause more severe structural damage in all the cell components investigated.     

Repair of functional and ultrastructural alterations after thermal injury of Physarum polycephalum

Repair of thermal injury of Physarum polycephalum Schw. plasmodia has been studied by light and electron microscopy. As a result of heating the plasmodia for 10 min at 42°C both the unordered and shuttle protoplasmic streaming were arrested; the outer plasmodial membrane showed alterations at sites of contact with water; the onset of the next mitosis was considerably delayed. The plasmodial ultrastructure was markedly disturbed, including disappearance of the granular component of the nucleoili and a compact, almost fibrillar structure of the latter. The mitochondria became distorted and their intracristal spaces enlarged while the outer and inner membranes appeared in some places to be separated. Glycogen particles disappeared from the cytoplasm. Recovery of both types of protoplasmic streaming of the motility of the plasmodium, of the resistance of its membrane to contact with water, and of the ability of the organism to go through the cell cycle went all hand in hand with the normalization of the structure of nucleoli, mitochondria and cytoplasm. All of the functional and structural characteristics are normalized within ca. 9 h following heating.     

The effects of cadmium exposure on the cytology and function of primary cultures from rainbow trout

Cultured epidermal cells from explants of skin of rainbow trout were used to study the cytological and functional changes following sublethal exposure to cadmium stress. The aim was to develop diagnostic markers for ecotoxicology. Cultures were exposed to the pollutant for 48 h. Cell structural and cytological changes were established by light and electron microscopy. Metabolic alterations were detected by immunohistochemistry. The relation between the initiation of cellular alterations and cadmium concentrations was compared in cultures exposed in commercially-available serum-free and serum-containing medium. The expression of stress proteins (metallothionein and heat shock protein) was also studied. Rainbow trout epithelial cells exposed to cadmium showed typical morphological changes indicative of cell death by apoptosis. Sublethal exposure also resulted in cellular metabolic disturbances with increased deposits of glycogen. Increased melanization was also observed. These changes appeared at lower concentrations of cadmium when cells were exposed in serum-free media than in serum-containing media. Cadmium induced the expression of heat shock proteins but not of metallothioneins. The results broadly confirm in vivo findings for cadmium toxicity and suggest that this in vitro technique may have applications in aquatic toxicology. © 1998 John Wiley & Sons, Ltd.     

Observations on the Responses of Lentil Root Cells to Hyphae of Fusarium oxysporum

The infection of lentil roots by Fusarium oxysporum Schlecht and the responses of the host cells to invading hyphae were examined by light microscopy. Hyphae from inoculum placed on the zone of cell elongation entered the roots at the juncture of epidermal cells within 8 h after inoculation. Although swollen hyphal apices were observed on the epidermal cells, root penetration occurred without formation of these structures or appressoria. The sheath of material found on the surface of uninoculated roots was absent from inoculated roots penetrated by hyphae. Prior to penetration, the epidermal cells became irregular in shape and their cytoplasm appeared to be plasmolysed or granular. Hyphae were observed in the cortex 10—12 h after inoculation and non–penetrated cortical cells were distinctly lobate. Often these lobed cells had a broad, peripheral band of diffuse cytoplasm. When hyphae were first observed in the cortical cells, the walls were ruptured and only slightly stained or unstained by toluidine blue. The inability of such walls to bind the stain may have been the result of the removal of wall components by fungal enzymes. Although extensive proliferation of hyphae was evident throughout the cortex after 24 h of incubation, the endodermis and vascular cylinder were free of hyphae for at least 72 h. Hyphae from inoculum placed on the root hairs or the root apex failed to penetrate the roots during the first 24 h of incubation. The cytological results herein are discussed in relation to the infection of field plantings by this pathogen.     

Rapid pacing of embryoid bodies impairs mitochondrial ATP synthesis by a calcium-dependent mechanism--a model of in vitro differentiated cardiomyocytes to study molecular effects of tachycardia

Tachycardia may cause substantial molecular and ultrastructural alterations in cardiac tissue. The underlying pathophysiology has not been fully explored. The purpose of this study was (I) to validate a three-dimensional in vitro pacing model, (II) to examine the effect of rapid pacing on mitochondrial function in intact cells, and (III) to evaluate the involvement of L-type-channel-mediated calcium influx in alterations of mitochondria in cardiomyocytes during rapid pacing. In vitro differentiated cardiomyocytes from P19 cells that formed embryoid bodies were paced for 24 h with 0.6 and 2.0 Hz. Pacing at 2.0 Hz increased mRNA expression and phosphorylation of ERK1/2 and caused cellular hypertrophy, indicated by increased protein/DNA ratio, and oxidative stress measured as loss of cellular thiols. Rapid pacing additionally provoked structural alterations of mitochondria. All these changes are known to occur in vivo during atrial fibrillation. The structural alterations of mitochondria were accompanied by limitation of ATP production as evidenced by decreased endogenous respiration in combination with decreased ATP levels in intact cells. Inhibition of calcium inward current with verapamil protected against hypertrophic response and oxidative stress. Verapamil ameliorated morphological changes and dysfunction of mitochondria. In conclusion, rapid pacing-dependent changes in calcium inward current via L-type channels mediate both oxidative stress and mitochondrial dysfunction. The in vitro pacing model presented here reflects changes occurring during tachycardia and, thus, allows functional analyses of the signaling pathways involved.     

Changes in cell ultrastructure and endogenous abscisic acid and indole-3-actic acid concentrations in Fatsia japonica leaves under polyethylene glycol-induced water stress

Ultrastructural alterations in epidermal and mesophyll cells as well as variations in bulk leaf endogenous ABA and IAA concentrations were studied in PEG-treated plants of Fatsia japonica Decne & Plank. Under stress induced by PEG vesicles containing fibrous material and electron-dense bodies associated with plasma membranes were observed. Cytochemical examination indicated that electron-dense bodies corresponded to lipids and the fibrous material of the vesicles were polysaccharides. Chloroplasts, mitochondria, nuclei and Golgi apparatus also showed modifications. A strong relationship was found between increasing PEG-induced water stress, increasing endogenous ABA and ultrastructural changes. In relation with leaf ontogeny and ABA concentration a higher ABA level was observed in younger than in older leaves. The differences in the endogenous concentrations of indole-3-acetic acid are unclear, except after 7 days of PEG-treatment. The increase in the endogenous abscisic acid concentration could be related with the ultrastructural changes.Abbreviations ABA = abscisic acid - IAA = indole-3-acetic acid - PEG = polyethylene glycol - = leaf water potential - TEM = transmission electron microscope     

Regions of lead uptake inLemna minor plants and localization of this metal within selected parts of the root

Investigations were carried out to determine the sites of lead uptake within the frond and the root ofLemna minor. With the sodium rhodizonate four regions favoured in lead uptake were distinguished: the frond region between the base and the node, the basal part of the root, and the regions at the proximal and distal ends of the root cap. For analysis in electron microscope only the root regions were chosen. The highest rate of lead uptake was found in the basal part of the root. Lead was present in the apoplast of this region after 5 min of exposure and was observed in the stelar cells after 30 min of incubation. Lead deposits were detected mostly in the cell walls adjacent to the plasma membrane and in the lumen of several endomembrane compartments - the endoplasmic reticulum (ER), dictyosomal vesicles, nuclear envelope and the vacuoles. Lead induced changes of cell ultrastructure; an increase in the number of membraneous structures, swelling of ER cisternae and distortion of the dictyosomal cisternae were observed after 2 to 6 h of exposure. We wish to thank Mrs. G. Winiecka for her technical assistance in preparing the photographs.     

Changes in the Endoplasmic Reticulum During Sieve Element Differentiation in Triticum aestivum

The changes in structure of the endoplasmic reticulum (ER) andits associations with other cell components have been studiedin differentiating protophloem sieve elements of root tips ofTriticum aestivum. In the young sieve elements single ER cisternaebearing ribosomes are dispersed in the cytoplasm. As differentiationprogresses ER increases in amount while a small proportion ofit aggregates into stacks or becomes associated with the nuclearenvelope and the mitochondria. These modifications occur inthe last two sieve elements containing ribosomes and coincidewith most dramatic changes in the degenerating nucleus. Stacksconsist of relatively few ER cisternae and may be encounteredfree in the cytoplasm or applied to the nuclear envelope. Electron-densematerial accumulates between the contiguous cisternae of thestacks. ER-attached ribosomes persist even in nearly maturesieve elements, but their pattern of arrangement becomes changed.The structural evidence indicates that only a few highly degradedER elements are retained in fully mature sieve elements. Triticum aestivum, root protophloem, sieve elements, endoplasmic reticulum, differentiation     

Stereo electron microscopy of the 25-nm chromatin fibers in isolated nuclei

The morphological effects of epidermal growth factor (EGF) on human carcinoma cells A-431 have been examined by scanning electron microscopy. These flat polygonal cells normally exhibit only small membrane folds, but show extensive ruffling and extension of filopodia within 5 min of exposure to EGF at 37 degrees C. This ruffling activity is transient, subsiding within another 5--15 min, but several other changes in surface morphology follow. Within the first hour of exposure to the hormone, the cell surface becomes exceedingly smooth and the nuclei seem to protrude above the plane of the otherwise thin monolayer, giving the cells a "fried egg" appearance. Cells at the edges of colonies gradually retract from the substrate, leading to reorganization, by 12 h, of the monolayer into multilayered colonies. EGF thus induces both rapid and long-term alterations in the morphology of these epidermoid cells.     

Changes in micropore system of roots of wheat and triticale seedlings under aluminum stress as determined using water vapor adsorption—desorption

Plant root structure including the pore system is severely injured in Al-toxic environments. Experimental water vapor desorption isotherms estimated using vacuum chamber method were used to characterize micropore build-up of roots of wheat (Lanca) and triticale (Debo) 4-day seedlings and its changes after 24 h (8 mg dm⁻³) aluminum stress. The micropore volumes and average pore radii increased after the stress whereas pore fractal dimensions decreased. The above changes were more pronounced for these roots which were not able to grow further after the stress when compared to the control roots, the total micropore volume was almost twice higher and the fractal dimension reached a value of 2 which is characteristic for flat, two-dimensional surface. The observed alterations of pore system indicate damage of root tissue and possible strong binding of aluminum on cell wall surfaces.     

Rapid pacing of embryoid bodies impairs mitochondrial ATP synthesis by a calcium-dependent mechanism—A model of in vitro differentiated cardiomyocytes to study molecular effects of tachycardia

Tachycardia may cause substantial molecular and ultrastructural alterations in cardiac tissue. The underlying pathophysiology has not been fully explored. The purpose of this study was (I) to validate a three-dimensional in vitro pacing model, (II) to examine the effect of rapid pacing on mitochondrial function in intact cells, and (III) to evaluate the involvement of L-type-channel-mediated calcium influx in alterations of mitochondria in cardiomyocytes during rapid pacing. In vitro differentiated cardiomyocytes from P19 cells that formed embryoid bodies were paced for 24 h with 0.6 and 2.0 Hz. Pacing at 2.0 Hz increased mRNA expression and phosphorylation of ERK1/2 and caused cellular hypertrophy, indicated by increased protein/DNA ratio, and oxidative stress measured as loss of cellular thiols. Rapid pacing additionally provoked structural alterations of mitochondria. All these changes are known to occur in vivo during atrial fibrillation. The structural alterations of mitochondria were accompanied by limitation of ATP production as evidenced by decreased endogenous respiration in combination with decreased ATP levels in intact cells. Inhibition of calcium inward current with verapamil protected against hypertrophic response and oxidative stress. Verapamil ameliorated morphological changes and dysfunction of mitochondria. In conclusion, rapid pacing-dependent changes in calcium inward current via L-type channels mediate both oxidative stress and mitochondrial dysfunction. The in vitro pacing model presented here reflects changes occurring during tachycardia and, thus, allows functional analyses of the signaling pathways involved.     

Spatial structure of mitochondria and ER denotes changes in cell physiology of cultured tobacco protoplasts

The structure of mitochondria and of the endoplasmic reticulum (ER) in mesophyll protoplasts and regenerated cells was studied in vivo using the dye DiOC₆(3) (3,3'-dihexyloxacarbocyanine iodide) and confocal laser scanning microscopy (CLSM). The relation to the cell's physiology was investigated using a hormone-based model system for elongation and division. The structure of the mitochondria and of their population depends on the status of the cell. In freshly isolated protoplasts small spherical mitochondria are clustered around the nucleus and the chloroplasts. During the first 4 days of culture they are transformed into long vermiform organelles which distribute evenly throughout the cytoplasm. In a medium containing only auxins, cells then enter a period of expansion. Their mitochondria retain the same structure but increase in quantity. In a medium with auxins and cytokinins cells start dividing. Their mitochondria typically become numerous and very small, and are distributed throughout the cytoplasm. Both types of organization were observed during weeks of ongoing expansion or division. The ER is always present as a network close to the cell membrane. In freshly isolated protoplasts a considerable part of the ER is clustered around the chloroplasts, the remaining part of the network has a reduced complexity and is partly broken. During subsequent protoplast culture the network grows into a complex web with fine meshes incorporating lots of plate-like structures. This is the case in elongating cells as well as in dividing cells. Finally, the ER looks similar to the structure found in epidermal cells of the intact plant.     

Superoxide production induced by short-term exposure of barley roots to cadmium, auxin, alloxan and sodium dodecyl sulfate

Key message

Abiotic stress-induced superoxide generation depending on its localization, level, duration and presumably also on the action of other signals may lead to different stress responses.

Abstract

The purpose of this study was to analyze the alterations in superoxide generation and morphogenesis following short-term Cd, IAA and alloxan treatments, during stress and recovery period in barley root tips. At low Cd concentration the transient accumulation of superoxide in the epidermal cells was accompanied by root growth inhibition and radial expansion of cortical cells in the elongation zone of root tips. These morphological changes were very similar to the externally applied IAA-induced responses. However, the role of superoxide generated in the epidermal cells by low concentration of Cd and IAA is probably alone not sufficient for the induction of these processes. SDS as an activator of NOX activity caused a strong accumulation of superoxide in the epidermal cells along the whole root apex but without any changes in root morphology and growth. On the other hand, higher Cd concentrations as well as alloxan stimulated the generation of superoxide in the cortical tissue of the elongation zone of root tip, which was accompanied by the induction of cell death. Our results suggest that enhanced superoxide generation, depending on its localization, level, duration and presumably also on the action of other signals, may lead to altered root morphology (15?μM Cd or IAA), root growth inhibition (alloxan), transient root growth cessation (30?μM Cd) or to the death of cells/root at higher (60?μM) Cd concentrations.     

Insulin protects against hepatic damage postburn

Burn injury causes hepatic dysfunction associated with endoplasmic reticulum (ER) stress and induction of the unfolded protein response (UPR). ER stress/UPR leads to hepatic apoptosis and activation of the Jun-N-terminal kinase (JNK) signaling pathway, leading to vast metabolic alterations. Insulin has been shown to attenuate hepatic damage and to improve liver function. We therefore hypothesized that insulin administration exerts its effects by attenuating postburn hepatic ER stress and subsequent apoptosis. Male Sprague Dawley rats received a 60% total body surface area (TBSA) burn injury. Animals were randomized to receive saline (controls) or insulin (2.5 IU/kg q. 24 h) and euthanized at 24 and 48 h postburn. Burn injury induced dramatic changes in liver structure and function, including induction of the ER stress response, mitochondrial dysfunction, hepatocyte apoptosis, and up-regulation of inflammatory mediators. Insulin decreased hepatocyte caspase-3 activation and apoptosis significantly at 24 and 48 h postburn. Furthermore, insulin administration decreased ER stress significantly and reversed structural and functional changes in hepatocyte mitochondria. Finally, insulin attenuated the expression of inflammatory mediators IL-6, MCP-1, and CINC-1. Insulin alleviates burn-induced ER stress, hepatocyte apoptosis, mitochondrial abnormalities, and inflammation leading to improved hepatic structure and function significantly. These results support the use of insulin therapy after traumatic injury to improve patient outcomes.     

Ultrastructure and movements of cell structures in normal pea and an ageotropic mutant

The pea mutant (Pisum sativum ageotropum) and the normal pea (P. sativum cv. Sabel) were compared in order to see if there were any differences in root anatomy or submorphology which could explain the presumed ageotropic behaviour of the mutant. In both types the root cap consists of a central core (columella) distinct from the peripheral part. The core contains five to six rows of columella cells, each consisting of 10 to 16 storeys of statocytes. The ultrastructure of the columella cells in the two types is very similar; the main difference is confined to the distribution of rough endoplasmic reticulum (ER), which in the mutant statocytes is evenly distributed throughout the cell, while in the normal pea statocytes it is mainly concentrated in the distal part at the “floor” of the cell. Using light micrographs, the movement of amyloplasts and nuclei have been followed in detail during a 40 min inversion period. The pattern of movement of the amyloplasts is apparently identical in the two types and the distances moved during the inversion period are 39 μm and 44 μm in the normal and mutant statocytes, respectively. The nucleus has not been observed to move in normal pea; a slight rearrangement of the nucleus position can be observed during the period 30 to 40 min after the start of inversion of the mutant. Based on magnified electron micrographs of the statocytes a morphometrical analysis was made of five cell structures – amyloplasts, nuclei, mitochondria, vacuoles and ER – which appeared to be freely movable or redistributable under the influence of the gravitational force.