Maintenance of air in intercellular spaces of plants

Although air-filled intercellular spaces are necessary and ubiquitous in higher plants, little attention has been paid to the possible mechanisms by which these spaces are kept from being flooded. The most likely mechanism is that the living plant cell may maintain a hydrophobic monolayer on the surfaces of adjacent intercellular spaces. The existence of `apparent free space' in cell walls and the fact that detergent solutions do not enter the intercellular spaces argue against this hypothesis. It is concluded that the mechanism by which these important air spaces are maintained is still unknown.     

Acid phosphatase activity in intercellular spaces in roots of Nymphoides peltata (S. G. Gmel.) O. Kuntze

Abstract. Acid phosphatase was found cytochemically in intercellular spaces in the root of Nymphoides peltata. Different methods, using lead salts and azo-dyes, gave similar results. Reaction product appeared on material, possibly cytoplasmic, within the intercellular spaces and also against the outer walls of cells which formed the intercellular spaces. Possible functions of acid phosphatase in intercellular spaces are discussed.     

Formation of intercellular gas space in the diaphragm during the development of aerenchyma in the leaf petiole of Sagittaria trifolia

The development of aerenchyma in the petiole of Sagittaria trifolia L. was studied by means of light-microscopy, scanning electron microscope, transmission electron microscope and immunofluorescence, focusing on the formation of intercellular spaces in diaphragms and its relationship with the organization of cortical microtubule arrays. A complex and organized honeycomb-like schizogenous aerenchyma formed by cylinders and vascular diaphragms was observed in the petiole of S. trifolia at different developmental stages. Cell division was the primary factor contributing to the increased volume of air spaces at early stages, while cell enlargement became the primary factor at later stages. The cortical microtubules localize at the sites where intercellular spaces and the secondary cell walls will be formed or deposited during the formation of intercellular spaces by the separation of diaphragm cells. Cortical microtubules were observed at the boundary of diaphragm cells and the fringes of intercellular spaces at later developmental stages where cell expansion occurs rapidly. These observations support the hypothesis that reorganization of cortical microtubule arrays might be related to the formation of air spaces in diaphragms and are involved in the deposition of secondary cell walls.     

Anatomic insights into the thigmonastic style tissue in Marantaceae

The irreversible style movement in Marantaceae is one of the quickest plant movements ever recorded. It is largely based on a special tissue construction, which is analysed herein for the first time. Histological sections, fluorescence and different electron microscopic techniques are used to study the style tissue of eight species in six genera of the Marantaceae. Tissue sections prior to the thigmonastic movement are only obtained after ether or chloroform narcosis. The tissue in the bending zone of the style has the construction of a lamellar collenchyma with elongated cells and extended intercellular spaces. The thicker cell walls abut the intercellular spaces, while the thinner walls between adjacent cells are extraordinarily porous. The tissue is thus characterised by strongly perforated cell bundles which are able to slide alongside each other and the intercellular spaces. Cell-wall loosening starts about 24 h before flowering and is interpreted as a result of enzymatic ageing processes. It is also found in other members of the Zingiberales, but only the Marantaceae take advantage of the maceration process. We conclude that the specific tissue construction contributes to the rapid style movement. It combines qualities for stabilisation (collenchyma), pliability (intercellular spaces) and water transport (extremely porous tissue), thus supporting the irreversible style bending by a sudden and final shift of fluid.     

Cavitation of intercellular spaces is critical to establishment of hydraulic properties of compression wood of Chamaecyparis obtusa seedlings

Background and Aims When the orientation of the stems of conifers departs from the vertical as a result of environmental influences, conifers form compression wood that results in restoration of verticality. It is well known that intercellular spaces are formed between tracheids in compression wood, but the function of these spaces remains to be clarified. In the present study, we evaluated the impact of these spaces in artificially induced compression wood in Chamaecyparis obtusa seedlings.Methods We monitored the presence or absence of liquid in the intercellular spaces of differentiating xylem by cryo-scanning electron microscopy. In addition, we analysed the relationship between intercellular spaces and the hydraulic properties of the compression wood.Key Results Initially, we detected small intercellular spaces with liquid in regions in which the profiles of tracheids were not rounded in transverse surfaces, indicating that the intercellular spaces had originally contained no gases. In the regions where tracheids had formed secondary walls, we found that some intercellular spaces had lost their liquid. Cavitation of intercellular spaces would affect hydraulic conductivity as a consequence of the induction of cavitation in neighbouring tracheids.Conclusions Our observations suggest that cavitation of intercellular spaces is the critical event that affects not only the functions of intercellular spaces but also the hydraulic properties of compression wood.     

Vegetative anatomy of subtribe Orchidinae (Orchidaceae)

Leaves in Orchidinae are essentially glabrous; anticlinal walls of foliar epidermal cells arc basically straight-sided to curvilinear, and cells arc fundamentally polygonal on both surfaces; adaxial cells are larger than abaxial cells. Stomata arc anomocytic and usually only abaxial and superficial; substomatal chambers are small to moderate; outer and inner guard cell ledges are mostly small. There is no hypodermis nor are there fibre bundles. Mesophyll is homogeneous, chlorcnchyma cells arc thin-walled, and intercellular spaces numerous. Crystalliferous idioblasts abound. Vascular bundles are collateral, organized in a single series. and lack associated sclerenchyma. Bundle sheath cells are thin-walled and chlorophyllous. Stems are glabrous; stomata arc frequent in one species, lacking in others. Cortical cells are oval to circular, thick-walled, and interspersed with triangular intercellular spaces. Ground-tissue cells are circular, and triangular intercellular spaces are present. Vascular bundles arc collateral and scattered throughout the ground-tissue or are absent from the central ground-tissue. Epidermis in absorbing roots is one-layered and non-velamcntous. Exodcrmal cells are thin-walled and dead cell walls bear tenuous scalariform bars; some species lack an exodermis. Outer cortical cells are polygonal and lack intercellular spaces; middle layer cortical cells are rounded with triangular intercellular spaces; inner layer cells are polygonal and lack intercellular spaces. Endodermis and pericycle are thin-walled and one-layered. Vascular cylinder is mostly 7–9-arch with xylcm and phloem components alternating regularly; vascular tissue is embedded in parenchyma; pith cells are parenchymatous, polygonal, thin-walled and lack intercellular spaces. Root tubers generally bear a velamen of variable thickness; bulbous-based unicellular hairs frequently form a dense mat; exodermal cells are thin-walled; dead cells have scalariform bars, passage cells are sparse. Ground-tissue consists of rounded water-storage and assimilatory cells interspersed with triangular or quadrangular intercellular spaces; peripheral cells arc polygonal lacking intercellular spaces. Vascular tissue consists of monarch to pentarch meristeles distributed thoughout the ground-tissue each surrounded by a uniscriale endodermis of thin-walled cells. Thin roots ofPlalanthera exhibit a typical central cylinder surrounded by a homogeneous cortex uninterrupted by meristeles; thicker roots show a central vascular cylinder and cortex in which meristeles are also present; in globoid root tubers there is no central cylinder, and the ground-tissue is replete with scattered meristeles. Because the central vascular cylinder in Platanthera gives rise to branches (meristeles), these represent components of a single vascular system and are not separate stelar entities as implied by the use of the term ‘polystele’.     

Analysis of the distribution of copper amine oxidase in cell walls of legume seedlings

Copper-containing amine oxidase (CuAO) has been proposed to play a role in H2O2 production in plant cell walls during cell development and in response to pathogen attack. We have compared the localisation of CuAO in pea (Pisum sativum L.), lentil (Lens culinaris M.) and chick pea (Cicer arietinum L.) grown under different light conditions, using both immuno- and histochemical techniques. The enzyme was detected by indirect immunofluorescence in the cell walls of parenchyma tissues of etiolated pea and lentil plants and was particularly abundant at intercellular spaces. Upon de-etiolation, CuAO largely disappeared from cortical cell walls except in the region of intercellular spaces. In the apical internode of light-grown seedlings, CuAO occurred mainly in cortical cell walls and, to some extent, in cell walls of xylem vessels. In both the elongation zone and mature regions of roots, CuAO was restricted to cortical cell walls and some cell junctions close to the meristem. Extensin epitopes co-localised to intercellular spaces of the cortex in de-etiolated pea, indicating that CuAO may have a role in cell wall strengthening at intercellular spaces. In chick pea, the localisation of the enzyme varied between different cultivars that have differing susceptibility to the fungus Ascochyta rabiei. In a susceptible cultivar Calia, immunogold labelling localised CuAO to cell walls of the cortex, as in lentil and pea, while in a resistant cultivar Sultano, it was most abundant in xylem vessels and, in light-grown plants, in the epidermis. These expression patterns are discussed with regard to the possible functions of amine oxidase in cell growth, cell differentiation and pathogen resistance.     

Sub-cellular location of H2O2, peroxidases and pectin epitopes in control and hyperhydric shoots of carnation

In carnation shoots (Dianthus caryophyllus cv. Killer), hyperhydricity was induced in in vitro culture using a low agar concentration. Using transmission electron microscopy, cytochemical techniques and immunolocation of JIM5 and JIM7 pectin epitopes, we followed the sub-cellular modifications of cell walls in relation to peroxidase activity and hydrogen peroxide accumulation during hyperhydricity induction. Peroxidase activity revealed a significant induction of the stomatal and epidermal cells as well as of the intercellular spaces of hyperhydric leaves. Similarly, hydrogen peroxide accumulated in the epidermal cell walls and the intercellular spaces of hyperhydric leaves. Immunolocation of an epitope recognised by the JIM5 antibody revealed the main unesterified nature of the cell walls. Such an epitope was located in the epidermal cell walls as well as in the corners of cell junctions in control leaves. However, hyperhydric leaves showed a total reduction of JIM5 labelling in the corners of cell junctions and a significant reduction of the intercellular spaces and the middle lamella. Highly-methylsterified pectin, recognised by the JIM7 antibody, was present to a slight extent in cell walls in control and hyperhydric leaves. We propose that the altered anatomy observed in hyperhydric carnation leaves could be regulated by the concomitant actions of pectin methyl esterases and free radicals, modifying the structure of the pectin and polysaccharides of the cell walls.     

Seed of Cercis siliquastrum L (Leguminosae,Caesalpinioideae): Reserves and their Utilization after Germination

An ultrastructural study on the reserves stored in cotyledons and endosperm and on their utilization after germination has been carried out in Cercis siliquastrum seeds. The observations have shown that all the cotyledonal reserves of proteins and lipids are used during seedling growth. However, inside the endosperm cells only the protein reserves are depleted, and large quantities of lipids remain undigested. In both cell walls and intercellular spaces of the endosperm cells huge masses of polysaccharide reserves are stored. However, only the polysaccharides inserted in the cell walls are dismantled, while those located in the intercellular spaces are not attacked by the lytic enzymes. Some hypotheses regarding the possible roles of those endospermic non-nutritional materials in the strategies of seed germination and seedling growth are suggested.     

Gas diffusion pathway in nodules ofCasuarina cunninghamiana

The gas diffusion pathway in nodules was traced by vacuum infiltration with India ink or aniline blue and by electron microscopy. India ink infiltration was observed in the outermost and the innermost cortex in sliced nodules, but not in intact nodules. With aniline blue infiltration, it was observed that intercellular air spaces in the outermost and the innermost cortex were connected to those in nodule roots. No air spaces were in contact with walls of infected cells, although intercellular air spaces existed in some groups of uninfected cells within the infected zone. Infiltration with either India ink or aniline blue could not be observed in the infected zone in essentially all cases. Thus it is suggested that the discontinuity of the intercellular air spaces represents a major resistance to O₂ diffusion in nodules ofCasuarina cunninghamiana.     

Distribution of the anionic sites in the cell wall of apple fruit after calcium treatment

Summary The ripening and softening of fleshy fruits involves biochemical changes in the cell wall. These changes reduce cell wall strength and lead to cell separation and the formation of intercellular spaces. Calcium, a constituent of the cell wall, plays an important role in interacting with pectic acid polymers to form cross-bridges that influence cell wall strength. In the present study, cationic colloidal gold was used for light and electron microscopic examinations to determine whether the frequency and distribution of anionic binding sites in the walls of parenchyma cells in the apple were influenced by calcium, which was pressure infiltrated into mature fruits. Controls were designed to determine the specificity of this method for in muro labelling of the anionic sites on the pectin polymers. The results indicate that two areas of the cell wall were transformed by the calcium treatment: the primary cell walls on either side of the middle lamella and the middle lamella intersects that delineate the intercellular spaces. The data suggest that calcium ions reduce fruit softening by strengthening the cell walls, thereby preventing cell separation that results in formation of intercellular spaces.Abbreviations EDTA ethylenediaminotetraacetic acid - PATAg periodic acid-thiocarbohydrazide-silver proteinate     

Intercellular pectic protuberances in Asplenium: new data on their composition and origin

BACKGROUND AND AIMS: Projections of cell wall material into the intercellular spaces between parenchymatic cells have been observed since the mid-19th century. Histochemical staining suggested that these intercellular protuberances are probably pectic in nature, but uncertainties about their origin, composition and biological function(s) have remained. METHODS: Using electron and light microscopy, including immunohistochemical methods, the structure and the presence of some major cell wall macromolecules in the intercellular pectic protuberances (IPPs) of the cortical parenchyma have been studied in a specimen of the Asplenium aethiopicum complex. KEY RESULTS: IPPs contained pectic homogalacturonan, but no evidence for pectic rhamnogalacturonan-I or xylogalacturonan epitopes was obtained. Arabinogalactan-proteins and xylan were not detected in cell walls, middle lamellae or IPPs of the cortical parenchyma, whereas xyloglucan was only found in its cell walls. Extensin (hydroxyproline-rich glycoproteins) LM1 and JIM11 and JIM20 epitopes were detected specifically in IPPs but not in their adjacent cell walls or middle lamellae. CONCLUSIONS: It is postulated that IPPs do not originate exclusively from the middle lamellae because extensins were only found in IPPs and not in surrounding cell walls, intercellular space linings or middle lamellae, and because IPPs and their adjacent cell walls are discontinuous.     

Extension Growth in Primary Cell Walls with Special Reference to Elodea canadensis

A quantitative study of distribution of perforations in thecellulose frame-work of longitudinal walls of cortical parenchymashows that their number and spacing change in an orderly manneras the tissue grow. These perforations are considered to bepit-fields, not primarily concerned with cell-wall growth, sincethey do nto occur in walls adjacent to intercellular spaces.The data are discussed in relation to the general problem ofcell extension, and are interpreted as showing that cell-wallgrowth is intersusceptive throughout; tip-growth does not occur.     

A study of the source of virus in the Pholem exudate appearing on leaves of Amscinckia infected with the beet curly top virus

Abstract Leaves of Amsinckia douglasiana discharging phloem exudate after infection with the beet curly top virus (BCTV) were studied with the electron microscope. Infected tissue differed from the noninfected in having much hyperplastic phloem characterized by abnormally high proportion of sieve elements, scarcity of companion cells, degenerating parenchyma cells, and some unusually large intercellular spaces. Many spaces contained amorphous debris. Particles resembling BCTV were discernible within the debris. Such particles were encountered also in the debris trapped between stomatal guard cells. Since the phloem exudate excreted from leaves of BCTV-infected plants contains virus particles, and since the virus is found extremely rarely in sieve elements, we suggest (1) that most of BCTV particles apparently released into intercellular spaces are derived from degenerating parenchyma cells in which the virus had multiplied; (2) that the exudate is derived from sieve elements of the hyper-plastic phloem in which the normal functional control by companion cells is lacking; (3) that the exudate leaks from the nontransporting sieve elements through cell walls into intercellular spaces and carries the virus to the outside. Initially, stomata may serve as exits for the infectious exudate, but subsequently ruptures in the epidermis are involved.     

Intercellular space and nonsynaptic interneuronal connections of the mammalian brain

Electron-microscopic investigations of various parts of the CNS in laboratory mammals and man have demonstrated that a single cerebral intercellular space, presenting an immediate internal medium for the nervous tissue elements, has different structure at the border separating the media. Here is a complex of structures, included into composition of the hemato-encephalic and liquor-encephalic barrier. Data of the literature on chemical composition of the intercellular contents and on specific membranous receptors make it possible to suppose a communication role of the intercellular spaces, ensuring nonsynaptic intercellular connections and modulation of the synaptic transmission. At the level of membranes, that make walls of the intercellular clefts by means of certain phylogenetically ancient humoral mechanisms, interaction of systems of synaptic and nonsynaptic interneuronal connections is secured.     

Anatomical studies of branchlet abscission related to crown modification in Quercus cerris L.

The basic anatomy of lateral twig insertion onto the main branch in both healthy and damaged Quercus cerris L. trees was studied. An abscission zone is always present: in healthy trees it is formed by a smaller number of cell layers than in damaged ones, where it is more evident with many layers of cells. Cells of the abscission zone are roundish, with many intercellular spaces between them; cell walls are thin, non-lignified and without secondary walls. No starch was found in cells of the abscission zone, where, instead, a few scattered calcium oxalate druses are seen.     

Natural deuterium and oxygen-18 enrichment in leaf water of cotton plants grown under wet and dry conditions: evidence for water compartmentation and its dynamics

Abstract. Significant differences in leaf water oxygen and hydrogen isotopic composition were observed between cotton plants grown under wet and dry conditions. The magnitude of the differences could be fully explained by the conventional model that describes the isotopic composition of an evaporating water pool under steady state conditions. The results indicate that leaf water isotopic composition is strongly influenced by transpiration rate via its effects on relative humidity adjacent to the leaf surface and on the isotopic composition of the air moisture. Our application of the model, however, provides evidence that leaf water must consist of a mixture of several isotopically distinct pools. These pools are suggested to reside in the symplast, in the cell walls and intercellular spaces and in the veins. A model is proposed suggesting that only the water residing in the cell walls and the intercellular spaces (the transpiration pool) interacts directly with the external environment. The large symplastic pool responds to the external environment to a limited extent via its relatively slow exchange with water in the transpiration pool. It is likely that the isotopic composition of water in the symplastic pool is strongly buffered against shortterm environmental variations, a possibility that would have important implications for the isotopic conditions under which organic matter biosynthesis occurs.     

Sinusoids--elements of the circulatory bed of the human heart

The literature concerning the problems of myocyrdial sinusoids has been analysed and morphological investigation of 98 human hearts has been performed. As demonstrates the comparison of the literature data, the term "sinusoid" in the human heart is applied to different structures. In some cases--to designate dilated areas of the venous part of the blood bed in the ventricular walls, in others--"hollow spaces", "intercellular spaces" and Vieussens--Thebesius vessels. Combining methods of intravascular injections and studying serial histological sections, it has been found that the sinusoids--the "hollow spaces", the "intercellular spaces", the "Veussens--Thebesius vessels" present the areas of intertrabcular spaces, which get into the planes of some sections, not being components of the myocardial blood bed. Spatial angioarchitectonics of the sinusoids--dilated areas in the venous part of the myocardial blood bed have been investigated by means of the plastic reconstruction method. It is suggested to apply the term "sinusoids" only to the given parts of the cardiac blood bed.     

THE ULTRASTRUCTURE AND DISPOSITION OF VESICULATED NERVE PROCESSES IN SMOOTH MUSCLE

The walls of the gastrointestinal tract and urinary bladder of rats were fixed in osmium tetroxide, embedded in methacrylate, and sectioned for electron microscopy. The examination of sections of smooth muscle tissue with the electron microscope reveals the presence of bundles of unmyelinated nerve fibers within the intercellular spaces. In addition, vesiculated nerve processes, bounded on their outer surfaces by delicate plasma membranes and typically containing varying quantities of synaptic vesicles and mitochondria, make intimate contact with the surface of smooth muscle cells. These nerve processes are similar in structure and disposition to nerve endings previously described in skeletal muscle, in the central nervous system, in peripheral ganglia, in receptors, and in glands. It is concluded that the relationships existing between vesiculated nerve processes and the surface of smooth muscle cells constitute neuromuscular junctions. Profiles of protrusions of smooth muscle cells are often seen protruding into the intercellular spaces. Here they occur singly or in groups, originating from one or more cells. Because of the plane of section the protrusions may sometimes appear as individual entities between the muscle cells. In such cases care must be exercised in their identification because they have characteristics similar to sectioned nerve processes which also occur in the intercellular spaces.     

Comparative Microscopical Studies on the Patterns of Calcium Oxalate Distribution in the Needles of Various Conifer Species

The distribution of calcium oxalate crystals in various conifer needles is visualized by light and electron microscopy. Such crystals occur (1) in the vascular bundle, either intracellularly in the xylem or phloem parenchyma, or extracellularly within the radial phloem walls; (2) extracellularly on the outside of the walls of mesophyll cells which face the intercellular spaces; (3) and finally as numerous small crystals within the cell walls of the epidermal cells, especially in the cuticular layer. The development and distribution of these apoplastic crystals is described in detail. Some hypotheses are finally presented for interpretations of these unusual patterns of the crystallization of Ca-oxalate outside the vacuole. Possible evolutionary aspects of this feature among the different conifer families are also discussed.