An Ultrastructural Study on the Development of Silicified Tissues in the Leaf Tip of Barley (Hordeum sativum Jess)

The development of silicified tissues in the tip of the fourthand fifth leaf of 7-week-old barley plants grown in nutrientsolution was investigated by transmission electron microscopy.The pattern of silicification was similar in the epidermal cells,sclerenchyma cells and intercellular spaces. Silica granulesfirst appeared around the periphery and eventually filled theentire cell lumen or space. The granules then polymerized toform a solid mass. The walls of the sclerenchyma cells alsobecame silicified but to a less extent than those of the epidermalcells, in which silicification sometimes occurred prior to thatin the lumen. The results are discussed in relation to the chemistry of silicaand possible mechanisms of silicification. Hordeum sativum Jess, barley, silica bodies, opalines, epidermal cells, sclerenchyma, ultrastructure     

Electron-probe Microanalysis Studies of Silica Distribution in Barley (Hordeum sativum L.)

Silicon occurence has been investigated by means of epidermalpeels, cryostat, and ultrathin sections of the internode, nodes,leaves, inflorescence bracts, and caryopsis of Hordeum sativumL. (cultivar Deba Abed) using the electron probe microanalyser.Analyses were made on growth stages during ear emergence andat maturity. The results indicate that silicon is present inthe internode with the highest concentration associated withthe opaline deposits. Detectable quantities are also found inthe outer tangential walls of the long cells, in the walls ofstomata, the sclerenchyma, and all vascular bundle regions.In mature upper internodes, silicifiation is confined to theupper third region, but this limit extends closer to the basalmeristem with increasing age of internode. The nodes have agreater concentration in the radial than in outer tangentialwalls. Heavy deposits are found in the leaves but with considerablevariation between blade and sheath, abaxial and adaxial surfaces,and the leaf position. The flag leaf contained the highest accumulations. In the inflorescence bracts (lemma and palea), silicon is detectableonly in the abaxial epidermis and hypodermis. Awns are alsoheavily silicified with the highest concentrations in the sclerenchymaand trichomes.     

Silicon Deposition in the Roots of Hordeum sativum Jess, Avena sativa L. and Triticum aestivum L.

Electron-probe microanalysis was used to investigate the locationof silicon at the proximal end of the seminal and adventitiousroots, of almost mature field-grown specimens of Hordeum sativumJess., Avena sativa L. and Triticum aestivum L. In the seminal roots silicon was confined to the endodermis,where it was present in the thickened inner tangential and radialwalls. The outer tangential walls also contained silicon inall of the cells in wheat and in occasional cells in barleyand oats. The adventitious roots of the three cereals displayed differencesin silicon deposition. In barley, silicon was present in allthe walls of the endodermal cells, whereas in oats it was onlylocated in the inner tangential and radial walls. Wheat showedcultivar differences, no silicon was detected in Capelle Desprez,but it was present in the thickened endodermis of Little Jossand Hustler. In all the samples studied silicon was absent fromthe sub-epidermal sclerenchyma layer. The results are discussed in relation to the possible functionsof the endodermis and the signficance of silicification. Hordeum sativum Jess, barley, Avena sativa L, oat, Triticum aestivum L, wheat, silicon deposition, electron-probe microanalysis     

Electron-probe Microanalysis Studies of Silicon in the Elongating Basal Internodes of Avena sativa (L.), Hordeum sativum (Jess.) and Triticum aestivum (L.)

Silicon deposits in the elongating basal internodes of almostmature, field grown specimens of Avena sativa, Hordeum sativumand Triticum aestivum were investigated using electron-probemicroanalysis. In A. sativa and H. sativum silicon was foundto be confined to the cells in the endodermal layer, being presentwithin the inner tangential and radial walls, and occasionallyin the outer tangential wall. In T. aestivum some silicon wasalso located in walls of cells surrounding the vascular bundles. The anatomy of the internodal tissues is discussed for the threespecies from light micrographs. The endodermal layer is discontinuousin A. sativa and H. sativum, in the former species it partlyextends around individual vascular bundles. In T. aestivum itforms a complete cylinder around the stelar region and alsoshows considerably more thickening of the inner tangential wallthan in the other species. The results are discussed in relation to the anatomy of theinternodal tissues and the possible function of silicon in theendodermis. Avena sativa L., Hordeum sativum Jess, Triticum aestivum L., oat, barley, wheat, silicon deposition, electron-probe microanalysis     

Silicon Deposition in the Roots, Culm and Leaf of Phalaris canariensis L

Silicon deposition in the roots, culm and leaf of canary grass(Phalaris canariensis L.) was investigated using light microscopy,scanning electron microscopy and electron probe microanalysis. In adventitious roots grown in solution silicon was concentratedin four endodermal walls. Silicon was not detected in the endodermisof aerial adventitious roots, but was present in the epidermisand outer cortical cell layers. Silicon deposition in the culm mainly took place in the epidermis,and particularly in epidermal papillae. The silica deposition pattern in the leaf was typical of thesub-group Festucoideae. The leaf blade showed deposits in costalprickle hairs and wavy rods, but few intercostal deposits. Inthe ligule deposition was confined to isolated groups of pricklehairs on the abaxial surface. The major sites of silica depositionin the leaf sheath were the stomatal subsidiary cells, papillaeand intercostal idioblasts. Prickle hairs were much less commonin the sheath than the blade, and costal wavy rods appearedto be absent in the sheath. Phalaris canariensis L., canary grass, silicification, root, culm, leaf, electron probe microanalysis     

Comparative anatomy and systematics of Catasetinae (Orchidaceae)

Catasetinae consist of five genera of pseudobulbous Orchidaceae of the Neotropics. Anatomy is characterized by sunken, three-celled foliar hairs, mostly tetracytic stomatal apparatuses, superficial stomata, homogeneous mesophyll, foliar fibre bundles, collateral vascular bundles in a single row, xylem and phloem sclerenchyma associated with vascular bundles in leaves, conical, and rough-surfaced silica bodies adjacent to vascular bundle sclerenchyma; epidermal cells of pseudobulbs with heavily thickened outer walls, pseudobulb ground tissue of assimilatory and water-storage cells, scattered vascular bundles in pseudobulbs, and sclerenchyma and stegmata associated only with phloem of pseudobulbs; roots with thin-walled velamen cells and tenuous spirals of cell wall material, distinctive epivelamen cells, thin-walled exodermal cells and vascular tissue embedded in parenchyma. Except for mucilaginous idioblasts that occur in Mormodes and Cycnoches , there are few outstanding anatomical differences among the five genera. Thus, there are few anatomical characteristics of phylogenetic value. The monophyly of Catasetinae is supported by the presence of sunken foliar hairs. Our results support a close relationship between Clowesia and Catasetum , and between Mormodes and Cycnoches. Among the outgroups Pteroglossaspis is especially distinctive.     

Intracellular Silica Deposition in Immature Leaves in Three Species of the Gramineae

The formation of solid, discrete deposits of opaline silicawithin the cell lumen of leaf tissues is reported in speciesrepresenting three subfamilies of the Gramineae; the preparationof a silica-minimal nutrient solution is discussed. Opal phytolithnumbers are related to tissue age and to two external silicaconcentrations for tiller leaves over a period of 32 days followingbud initiation. Variations in silica deposition patterns among the individualleaves of a homologous series on the shoot apex are relatedto differential growth-rates. During ontogeny, deposition occursin an ordered sequence of cell types related to the basipetalmaturation gradient within the leaf. Initial deposition wasdetected in silica cells (idioblasts) of imbricated, bud leavesof Sieglingia decumbens, 1 mm long; phytoliths are confinedto these cells in expanding, basal portions of the leaf. Phytolithcounts/sq mm of epidermis vary with the degree of long-cellexpansion. The error is reduced by expressing silicificationas the number of phytoliths/100 silica cells. Post-expansiondeposition was initiated in epidermal long cells of the leafblade tip. The higher silica concentration resulted in a morerapid utilization of available deposition sites and larger phytoliths(P = 0.001). A passive cell influx of silicic acid is discussedin relation to cytodifferentiation.     

Culm strength of barley : correlation among maximum bending stress, cell wall dimensions, and cellulose content

Grass culms are known to differ in breaking strength, but there is little physicochemical data to explain the response. The fourth internode of four brittle and two nonbrittle barley (Hordeum vulgare L.) strains were used for physical and chemical studies of culm strength. Inner and outer culm diameters of brittle strains (3.6 ± 0.2 and 5.0 ± 0.1 millimeters) were not significantly different from those of nonbrittle strains (3.9 ± 0.2 and 5.2 ± 0.2 millimeters). Maximum bending stress, at which the culm was broken, was 192 ± 34 g/mm² for brittle and 490 ± 38 g/mm² for nonbrittle strains. Wall thickness and cell dimensions of epidermal, sclerenchyma, and parenchyma cells were measured in culm cross sections. The area of cell wall per unit cell area for each tissue was significantly correlated with the maximum bending stress (r = 0.93 for epidermis, 0.90 for sclerenchyma, and 0.84 for parenchyma). Cell walls of brittle culms had 6 to 64% as much cellulose content as those of nonbrittle culms. Maximum bending stress correlated significantly with cellulose content of the cell walls (r = 0.93), but not with the contents of noncellulosic compounds. The lower cellulose content of the brittle culm was significantly correlated with brittleness.     

Biogenic Opal in Three Species of Gramineae

Particulate biogenic opaline silica is concentrated in cellwalls, intercellular deposits and cell lumina of all portionsof the above-ground plant body of three species of PanicoidGramineae,Andropogon gerardi, Sorgastrum nutans and Panicumvirgatum. Morphologically distinct opal phytoliths form notonly in long cells, short cells, trichomes, stomatal elementsand bulliform cells of the epidermis but also within the cellularstructure of mesophyll, vascular, and sclerenchyma tissues.Roots and rhizomes contain measurable quantities of opalinesilica, and phytoliths develop in epidermal long cells, saddle-shapedshort cells, vascular cells, and intercellular deposits. A morphologicallyunique plate-phytolith, formed by silicification of the innertangential wall of the endodermis, is present in the roots ofall three species. Differences in the quantity of opaline silicaoccur between species and between parts of the same species.The amount of opal deposited in the soil annually by root systemsand above-ground parts is approximately equal in magnitud Andropogon gerardi, Sorgastrum nutans, Panicum virgatum, opaline silica deposition     

Isolation of a novel cell wall architecture mutant of rice with defective Arabidopsis COBL4 ortholog BC1 required for regulated deposition of secondary cell wall components

The plant secondary cell wall is a highly ordered structure composed of various polysaccharides, phenolic components and proteins. Its coordinated regulation of a number of complex metabolic pathways and assembly has not been resolved. To understand the molecular mechanisms that regulate secondary cell wall synthesis, we isolated a novel rice mutant, cell wall architecture1 (cwa1), that exhibits an irregular thickening pattern in the secondary cell wall of sclerenchyma, as well as culm brittleness and reduced cellulose content in mature internodes. Light and transmission electron microscopy revealed that the cwa1 mutant plant has regions of local aggregation in the secondary cell walls of the cortical fibers in its internodes, showing uneven thickness. Ultraviolet microscopic observation indicated that localization of cell wall phenolic components was perturbed and that these components abundantly deposited at the aggregated cell wall regions in sclerenchyma. Therefore, regulation of deposition and assembly of secondary cell wall materials, i.e. phenolic components, appear to be disturbed by mutation of the cwa1 gene. Genetic analysis showed that cwa1 is allelic to brittle culm1 (bc1), which encodes the glycosylphosphatidylinositol-anchored COBRA-like protein specifically in plants. BC1 is known as a regulator that controls the culm mechanical strength and cellulose content in the secondary cell walls of sclerenchyma, but the precise function of BC1 has not been resolved. Our results suggest that CWA1/BC1 has an essential role in assembling cell wall constituents at their appropriate sites, thereby enabling synthesis of solid and flexible internodes in rice.     

SCANNING ELECTRON MICROSCOPY AND ELECTRON MICROPROBE ANALYSIS OF SILICIFICATION PATTERNS IN INFLORESCENCE BRACTS OF AVENA SATIVA

Using scanning electron microscopy, we determined the kinds and distribution of epidermal cell types in Avena inflorescence bracts (glume, lemma, and palea). Electron microprobe analysis of silica deposition in these epidermal cells showed that silica cells constitute one of the important deposition sites. Probe ratio data indicate that the silica deposited is 74 % pure. Significant amounts of silica also become deposited in the trichomes and lesser amounts in the walls of long epidermal cells. None could be detected in the stomata. The possible functional significance of silica deposition in epidermal cells of these bracts is discussed.     

Comparative vegetative anatomy of Stanhopeinae (Orchidaceae)

Stanhopeinae are a group of tropical American orchids characterized by euglossine bee pollination and lateral inflorescences stemming from the bases of pseudobulbs. Leaves are hypostomatal, and all stomatal configurations are tetracytic. Chlorenchyma is homogeneous and characterized by fibre bundles in adaxial/abaxial or adaxial/median/abaxial positions. Collateral vascular bundles occur in a single row and feature phloic and xylic sclerenchymatous caps and thin-walled bundle sheath cells. Fibre bundles and vascular sclerenchyma are accompanied by stegmata containing conical silica bodies. Pseudobulbs have thick-walled turbinate epidermal cells and ground tissue of smaller, living assimilatory cells and larger, dead water-storage cells. Fibre bundles are usually absent but occur in several genera. Collateral vascular bundles show phloic sclerenchyma, but xylic sclerenchyma occurs only in die larger vascular bundles. Phloic and xylic sclerenchyma are associated with stegmata containing conical silica bodies. Roots are velamentous. Velamen cell walls have fine, spiral thickenings. Exodermal cells are thin-walled. The cortex features scattered thick-walled cells and in some cases branched bars of secondary cell wall material. Endodermis is either u-or O-thickened, but pericycle is always O-thickened opposite the phloem. Vascular tissue consists of alternating strands of xylem and phloem surrounded by a matrix of thick-walled cells. Pith cells may be parenchymatous or sclerenchymatous.     

Silicon in Imperata cylindrica (L.) P. Beauv: content,distribution, and ultrastructure

Silicon concentration, distribution, and ultrastructure of silicon deposits in the Poaceae Imperata cylindrica (L.) P. Beauv. have been studied. This grass, known for its medicinal uses and also for Fe hyperaccumulation and biomineralization capacities, showed a concentration of silicon of 13,705?±?9,607 mg/kg dry weight. Silicon was found as an important constituent of cell walls of the epidermis of the whole plant. Silica deposits were found in silica bodies, endodermis, and different cells with silicon-collapsed lumen as bulliforms, cortical, and sclerenchyma cells. Transmission electron microscope observations of these deposits revealed an amorphous material of an ultrastructure similar to that previously reported in silica bodies of other Poaceae.     

Silica Distribution in the Caryopsis and Inflorescence Bracts of Foxtail Millet [Setaria italica (L.) Beauv.] and its Possible Significance in Carcinogenesis

Silicon deposits in the caryopsis and inflorescence bracts offoxtail millet [Setaria italica (L.) Beauv.] were investigatedusing light microscopy, scanning electron microscopy and electronprobe microanalysis. The samples were obtained from Lin Xian,Henan province, northern China and CSIRO, Australia. High concentrations of silicon were observed in the papillaeon the external surfaces of the inflorescence bracts, and inthe epidermal cells. In the caryopsis silicon was depositedin the aleurone layer. Silicification was heavier in the LinXian samples. The heavy accumulation of silica in the foxtail millet and thepossibility that fragments of plant silica may be implicatedin the aetiology of oesophageal cancer in the Lin Xian regionare discussed. Setaria italica (L.), Beauv, foxtail millet, caryopsis, silica distribution, scanning electron microscopy     

Comparative vegetative anatomy and systematics of Vanilla (Orchidaceae)

Vanilla is a pantropical genus of green-stemmed vines bearing clasping (aerial) and absorbing (terrestrial) roots. Most vanillas bear normal, thick foliage leaves; others produce fugacious bracts. Seventeen species, including both types were studied. Foliage leaves of Vanilla are glabrous, have abaxial, tetracytic stomatal apparatuses, and a homogeneous mesophyll. Species may or may not have a uniseriate hypodermis. Crystals occur in the foliar epidermises of some species, but all species have crystalliferous idioblasts with raphides in the mesophyll. Vascular bundles in leaves are collateral and occur in a single series alternating large and small. Sclerenchyma may or may not be associated with the vascular bundles. Scale leaves may be crescent or C-shaped and usually have abaxial stomatal apparatuses. A hypodermis may or may not be present; the mesophyll contains raphide bundles in idioblasts. Vascular bundles are collateral and occur in a single row sometimes aligned close to the adaxial surface. They may or may not be associated with sclerenchyma. Stems of leafy vanillas show a sclerenchyma band separating cortex from ground tissue; stems of leafless vanillas do not show a sclerenchyma band. Ground tissue of the stem may consist solely of assimilatory cells or mixed assimilatory and water-storage cells. In some species centrally located assimilatory cells are surrounded by layers of water-storage cells. A uniseriate hypodermis is present in all stems. Sclerenchyma may completely surround the scattered collateral vascular bundles, occur only on the phloem side, or be absent. Both aerial and terrestrial roots are notable for their uniseriate velamen the cell walls of which may be unmarked or ornamented with anticlinal strips. Exodermis is uniseriate; the cells vary from barely thickened to strongly thickened. Only the outer and radial walls are thickened. Cortical cells of aerial roots generally have chloroplasts that are lacking from the same tissue of terrestrial roots. Raphide bundles occur in thin-walled cortical idioblasts. Endodermis and pericycle are uniseriate; pericycle cells are all ?-thickened opposite the phloem. Cells of the endodermis are either ?- or ∪-thickened opposite the phloem. Vascular tissue may be embedded in thin- or thick-walled sclerenchyma or in parenchyma. Metaxylem cells are always wider in terrestrial than in aerial roots of the same species. Pith cells are generally parenchymatous but sclerotic in a few species.     

Silicon Distribution and Anatomy of the Grass Rhizome, with Special Reference to Miscanthus sacchariflorus (Maxim.) Hackel.

Rhizome anatomy is described for Miscanthus sacchariflorus (Maxim.)Hackel. Solid silica deposits, detected as elemental siliconby electron-probe microanalysis and energy-dispersive X-rayanalysis, are confined to cell walls of three concentric zonesconsisting of the uniseriate epidermis, and parenchyma layersaround the cortical air lacunae, and the central cavity, respectively.Si is localized in outer tangential walls of the epidermis,while occurring in all walls of nucleated, parenchyma cellsforming the two internal zones. In comparison, the root exhibitsonly one Si zone. Rhizome Si distribution more closely resemblesthat for Phragmites australis, than for related members of theAndropogoneae. P. australis similarly exhibits aerenchyma anda central cavity. Thus, internal anatomy may strongly influencesilicon distribution. A comparison of taxa of four tribes indicatesthat epidermal wall deposition is common, followed by specificinternal localization in up to three zones of perivascular tissues. Silicon accumulation occurs early in the epidermis of the youngapex of M. sacchariflorus, decreasing sharply across an internodetransection. In comparison, the oldest, basal internodes exhibitvery high Si X-ray counts in each of the three zones, the highestoccurring in the most internal zone around the central cavity.Early Si mobilization in the rhizome apex may resist shearingand abrasion during horizontal growth extension, while depositsbordering aerenchyma of older internodes may resist compression. Miscanthus sacchariflorus (Maxim.) Hackel, plume grass, rhizome, silicification, anatomy, aerenchyma     

The taxonomic value of floral characters in Rapateaceae (Poales-Monocotyledons)

The floral anatomy of Cephalostemon, Monotrema, Rapatea, Spathanthus, and Stegolepis was studied for taxonomic purposes. All species studied share colleters between the floral parts; sepals, petals, anthers, and style covered by an ornamented cuticle; short epidermal cells with sinuous walls on the abaxial surface of the petals; tetrasporangiate anthers with phenolic idioblasts in the epidermis; endothecium with spiral thickenings; incompletely septate ovary; and anatropous, bitegmic ovules. The floral anatomy is useful not only for characterizing the family, but also for delimiting the subfamilies and genera. Sepals with silica bodies in the epidermal cells; mature anther wall composed of epidermis, endothecium, and middle layer; absence of phenolic idioblasts in the sepals, filaments, and ovary; and stylar epidermal cells with thickened external periclinal wall support Rapateoideae. Cephalostemon and Rapatea show a great number of similarities, corroborating their close relationship indicated in the phylogenetic analyses of the family. Monotrema shares few characters with the genera of Rapateoideae, corroborating its placement in Monotremoideae. Stegolepis shows several distinctive characters, probably related to the greater diversity found in this genus.     

Structure and Function of Silica Bodies in the Epidermal System of Grass Shoots

Silica (SiO₂.nH₂O) is deposited in large quantities in the shootsystems of grasses. In the leaf epidermal system, it is incorporatedinto the cell wall matrix, primarily of outer epidermal walls,and within the lumena of some types of epidermal cells. This biogenic silica can be stained specifically with methylred, crystal violet lactone, and silver amine chromate. At theultrastructural level, the silica in lumens of silica cells,bulliform cells and long epidermal cells is made up of rodsabout 2.5 µm in length and 0.4µm in width. Ultimateparticles in the rods range from 1 to 2 nm in diameter. In contrast,silica in the cell wall matrix of trichomes and outer wallsof long epidermal cells is not rod-shaped, but rather, formsroughly spherical masses. Detailed analyses are presented on the frequencies of occurrenceof the different types of epidermal cells that contain silicain the leaves of representative C₃ and C₄ grasses. The C₄ grasseshave higher frequencies of bulliform cell clusters, silica cells,and long epidermal cells, whereas the C₃ grasses have higherfrequencies of trichomes. No correlation was found in the frequencyof occurrence of silica bodies in bulliform cells for C₃ grassesas compared with C₄ grasses. Of all the grasses examined, Coix,Oryza, and Eleusine had the highest densities of such bodies,and some taxa had no silica bodies apparent in their bulliformcells. The idea that silica bodies in bulliform cells and silica cellsmight act as "windows’ and trichomes might function as‘light pipes’ to facilitate light transmission throughthe epidermal system to photosynthetic mesophyll tissue belowwas tested. The experimental data presented do not support eitherof these hypotheses. C₂ and C₄ grasses, biogenic silica, light pipes, window hypothesis, silica staining, silica ultrastructure     

Water Pathways in Higher Plants: I. FREE SPACE IN WHEAT LEAVES

A technique has been developed for the study of pathways ofwater movement in the xylem and free space of wheat leaves.Plants were treated with Lead-EDTA chelate through either theroots or the leaves; after treatment the lead was precipitatedin situ as lead sulphide with hydrogen sulphide gas and itslocation determined by light and electron microscopy. Bulk water movement was in the lumen of the xylem, where therewas always a heavy deposit of lead sulphide after root treatments.Outside the xylem the deposits were confined to the cell wallsand were most dense in the middle lamella. Deposits were notfound in the cells themselves. The main zones of water loss,marked by heavy deposits of lead sulphide, were associated withthe stomata, the junctions of the periclinal walls of the epidermalcells, and the cuticle, leaf hairs, and specialized epidermalcells with pitted walls associated with the vascular bundles. Entry of lead chelate into the leaves was adequately describedby a diffusion model. The free space seemed to be located mainlyin the water of hydration of the pectin middle lamella and wasesmated to occupy 3 to 5 per cent of the volume of the tissue.