Scanning Electron Microscopy of Silica Deposits in the Culms, Floral Bracts and Awns of Barley (Hordeum sativum Jess.)

Deposits of silica in the culm internodes, floral bracts andawns of Hordeum sativum Jess (cv Deba Abed) have been investigatedusing the scanning electron microscope The deposits were isolatedfrom all organic matter by digestion with nitric and perchloricacids Two basic types of deposits were recognized, lumen andwall silicification, the latter with or without lumen infillings In the culm internodes, lumen deposits are derived from idioblasts(‘hats’), sclerenchyma and xylem vessels In thefloral bracts they are derived from idioblasts (‘hats’and astenform opals), epidermal long cells (dendriform opals),sclerenchyma and xylem vessels The shape of these deposits doesnot generally resemble the outline of the cell itself, but depositsderived from cell walls do closely resemble the infact cell.In the culm, the walls of stomatal cells, trichomes and, largelythe outer tangential walls of mature long epidermal cells, becomesilicified, together with some cork cells In the floral bracts,silica is found in most epidermal cell walls but is confinedto the trichomes, scutiform cells and costal epidermal cellsearly in their development At maturity concentrations of silicaare much higher in the floral bracts and awns than in the culmsand leaves The results are discussed in relation to the pattern of depositionand its possible functions. Hordeum sativum Jess, barley, silica deposits, opals, scanning electron microscopy     

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     

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     

Distribution of Silicified Cells in the Leaf Blades of Pleioblastus chino(Franchet et Savatier) Makino (Bambusoideae)

Distribution of silicified cells in the leaf blades of Pleioblastuschino was investigated using a light microscope and a scanningelectron microscope equipped with an energy dispersive X-raymicroanalyser. The most dense accumulation of silica was foundin epidermal tissues. Little silica was deposited in vascularbundles and chlorenchyma, while more was deposited in bundlesheath and fusoid cells. In the epidermis, silica density andfrequency of silicified cells differed depending on cell type,although silica deposition was observed in most cell types.Heavy deposition was found in silica cells, bulliform cells,micro hairs and prickle hairs. Silica cells were the cell typemost frequently silicified (96.9–99.7%) in the adaxialand abaxial epidermis. Silica may be deposited as leaf tissuesage.Copyright 2000 Annals of Botany Company Pleioblastus chino(Franchet et Savatier) Makino, bamboo, silicified cells, leaf blade, epidermis, chlorenchyma, silica, clearing method, freeze-fracturing, freeze-drying, light microscopy, scanning electron microscopy, X-ray microanalysis     

Silicification of the adaxial epidermis of leaves of a panicoid grass in relation to leaf position and section and environmental conditions

Many studies relate silica content in plants with internal or external factors; however, few works analyse the effect of these factors on the silicification of different cell types. In this study, we examined the effect of leaf section and leaf position, and environmental conditions on the percentages of silicified epidermal cells of a native Pampean panicoid grass, Bothriochloa laguroides D. C. Pilger. Two different environmental situations were selected for the collection of plants: a natural wetland and a quartzite quarry, located in the southeast Buenos Aires province, Argentina. Clarification and staining methodologies were applied so as to study the distribution of silicified cells in different sections of leaves of the plants collected. Two and three-factor anovas were applied to the data. Between 13% and 19% of total cells of the adaxial epidermis of leaf blades were silicified. Typical silica short cells were the largest contributor to total silicified cells (53-98%), while the second largest contributor was bulliform cells (0-30%). Percentages of total silicified cells were higher in superior than in inferior leaves, while values from leaf sections varied. When collection sites were compared, plants growing in Los Padres pond, where the silica content in soils is higher, had the higher percentage of silicified cells. Among all types of cell, bulliform cells showed differences in the proportion of silicified cells between leaf position and section and collection site. These results show that silica availability in soils is an important factor that conditions silica accumulation and overlaps with the transpiration effect.     

The Role of Extracellular Polymeric Substances in the Silicification of Calothrix: Evidence from Microbial Mat Communities in Hot Springs at Yellowstone National Park,USA

We provide nanoscale evidence of the role of sheath exopolymers in the silicification of the sheathed cyanobacteria Calothrix. Electron microscope observations of silicified Calothrix cells revealed that silica accretes directly onto EPS sheath fibrils to produce an open web of silica particles that could remain permeable to nutrients and waste products. We also found that silicified Calothrix cells from different microhabitats contained morphologically distinct silica particles. Differences in silicification texture suggest that environmental variables may influence silicification at the nanoscale. We develop a framework based on aggregation kinetics to address silicification processes in Calothrix and other sheathed cyanobacteria.     

Electron-probe Microanalysis Studies of Silicon in the Epicarp Hairs of the Caryopses of Hordeum sativum Jess., Avena sativa L., Secale cereale L. and Triticum aestivum L.

Silicon deposits in the epicarp hairs of the caryopses of mature,field-grown specimens of Hordeum sativum, Avena sativa, Secalecereale and Triticum aestivum were investigated using electron-probemicroanalysis. In all four cereals, silicon was most concentratedat the extreme tips of the hairs. In barley, it was the onlyelement detected; in the other three cereals potassium and calciumwere located below the tip. In wheat, chlorine was also detected. The hair bases of the different cereals displayed variationin the elements detected. Silicon and polassium were presentin all four and calcium present in all except rye. The hairbases of wheat also contained chlorine; phosphorus and zincwere located in barley. The latter alone showed variation ofelements between hair bases. Scanning electron microscopy revealed heavy striations of thehair tips of barley and oats. In rye and wheat, the tips weresmooth but there were slight surface markings below the tip. The results are discussed in relation to the possible functionsand significance of the epicarp hairs and their silicification. Avena sativa L., Hordeum sativum Jess., Secale cereale L., Triticum aestivum L., oat, barley, rye, wheat, epicarp hairs, silicon, electron-probe microanalysis, scanning electron microscopy     

An Ultrastructural Study on the Developmental Phases and Silicification of the Glumes of Phalaris canariensis L.

In the glume of Phalaris canariensis L. silicon deposition takesplace in the macrohairs, papillae, prickle hairs and silicacells of the abaxial epidermis before panicle emergence. Early in their development the macrohairs have large vacuolesand thin walls. At maturity the walls become thickened and aremajor sites of silica deposition. Dry ashing reveals a helicalpattern within the hair walls. Distinct papillae and prickle hairs were first observed oneweek before panicle emergence. Here silicification was initiallyconfined to outer tangential walls, but by two weeks after emergencetheir cytoplasmic contents had broken down, and the lumina werefilled with siliceous granules. Cork-silica twin cells werealso present in the abaxial epidermis. By panicle emergencethe silica cells were infilled, but the cork cells retainedtheir cytoplasmic contents. The long cells of the abaxial epidermiswere initially thin walled, but thickening occurred in the outertangential wall, this being complete by one week after emergence.These cells remained relatively unsilicified throughout. After panicle emergence the adaxial epidermal cells, and theirassociated parenchyma cell layers began to lose their cellularcontents and collapse. This process was complete two weeks afteremergence when the collapsed walls formed a thin internal layerbetween the two epidermi. Electron opaque granular material,containing several elements, but predominantly calcium, waspresent between the collapsed cell walls. The results are comparedwith those for the lemma, and silica deposition mechanisms arealso discussed. Phalaris canariensis L., canary grass, silicification, trichome, glume, ultrastructure     

Endolithic micro-organisms silicification of a bivalve fauna from the Silurian of Gotland

The silicified Wenlockian bivalve shells at Möllbos have been fragmented to a considerable extent. Shells which were broken prior to silicification exhibit possible original shell layers while those which were fragmented during laboratory treatment show no primary structures. The fauna at Möllbos was attacked by endolithic micro-organisms. The borings of these were then coated with a carbonate envelope. After burial the unattacked original shell material was dissolved the envelope silicified. Later, empty moulds were subsequently filled with drusy calcite occasionally with quartz crystals. A third silicification went occurred at a later diagenetic stage when matrix had become lithified.     

微生物硅化作用模拟实验

硅化保存微体化石结构的识别和鉴定一直存在争议,现代人工模拟生物硅化实验有助于对硅化作用保真度(silicification fidelity)等问题的理解。本实验通过人工配置浓度为1000ppm的硅溶液模拟自然硅化环境,观察杆状细菌(bacillus)在该环境中的早期硅化作用。实验发现这种细菌可以在该硅化环境中快速硅化,并较好地保存其形态。硅化细菌表面硅质微颗粒(nano-spheres)沉积明显分两层,紧密规则的内层硅质微颗粒较好地保存了细菌表面结构,外层硅质微颗粒仅保存细菌轮廓。实验结果期望能为硅质细菌类微体化石的鉴别提供有益参考,并为其成因探讨提供更多实证依据。     

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.     

A Developmental Study of Silicification in the Trichomes and Associated Epidermal Structures of the Inflorescence Bracts of the Grass, Phalaris canariensis L

Silicon accumulation is greatest in the abaxial epidermis ofthe inflorescence bracts of Phalaris canariensis L. It occursinitially in glumes 1 week before emergence, at low levels inthe trichomes and long cells, but at high levels in the papillae.Accumulation in long cells remains low throughout, althoughit increases with time. Papillae and prickle hair silicon countsconsiderably exceed values for other epidermal structures. Siliconis also deposited in glume macrohairs before emergence, andat maturity reaches intermediate levels. Little silicon is present in the macrohairs and long cells ofthe abaxial epidermis of the fertile lemma prior to emergence.After emergence, however, silicon is rapidly accumulated inthese structures, and at maturity they have the highest siliconlevels of all the mature bract tissues. Silicon accumulation in the lemma lags behind that of the glumeuntil emergence, when it increases more rapidly, and by maturityexceeds that of the glume in macrohairs and long cells. Thesignificance of the results is discussed in relation to wallthickening, bract anatomy and panicle emergence. Phalaris canariensis, L, canary grass, inflorescence bracts, silicification, trichome, glume, lemma     

Phytoliths in woody plants from the northern slope of the Changbai Mountain (Northeast China), and their implication

Phytoliths were extracted from 14 woody plants collected on the northern slope of the Changbai Mountain, including 10 broad-leaved species and 4 conifers. A total of 14 morphotypes of phytoliths were identified, including 3 types first examined in this study. Phytoliths in broad-leaved species were mostly silicified epidermal cells, cell walls, and vascular tissues; phytoliths in conifers were mainly silicified epidermal cells, cell walls, hypodermal cells, and parenchyma cells. Phytoliths produced by broad-leaved species in this region were usually not well silicified, and were fragile, whereas those produced by conifers were better silicified; this might be because of the different lengths of the growth periods. Phytoliths were found have potential in studies of fluctuations of the tree line in this region, and this study also provided a reference for further study of phytoliths in this region and the regional contrast of phytolith assemblages.     

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     

A Comparison of Nitrate-selective Microelectrodes made with Different Nitrate Sensors and the Measurement of Intracellular Nitrate Activities in Cells of Excised Barley Roots

Nitrate-selective microelectrodes based on a number of nitratesensors were compared. The electrode properties tested includedlog-linear slope of the calibration curves, detection limit,and ion selectivity. The nitrate sensor mixture described inan earlier paper performed favourably when compared with othernitrate-selective mixtures or with commercially-available macroelectrodes.This earlier mixture consisted of 6% methyltridodecylammoniumnitrate, 65% n-phenyloctyl ether, 23% poly(vinylchloride), 5%nitrocellulose, and 1% methyltriphenyl phosphonium bromide.These electrodes, even when stored backfilled remained nitrate-selectivefor several days although there was eventually some deteriorationin performance. The electrodes were used in vitro for assaying nitrate in barleyroot extracts and a linear relationship was found between resultsfrom ion chromatography and microelectrode measurements. Intracellularmeasurements made in vivo in epidermal cells of excised barleyroots identified two populations of measurements believed tobe the cytoplasm and the vacuole. Significant decreases in compartmentalnitrate activities were measured 2 to 6 h after excision. Theseresults indicate that the nitrate pool in both the cytoplasmand vacuole of root epidermal cells is sensitive to root excisionand question the physiological significance of measurementsmade on excised roots. Key words: Nitrate-selective microelectrodes, barley root, compartmentation, nitrate     

Ficus carica L. leaf anatomy: Trichomes and solid inclusions

Ficus carica L., a typical plant of the Mediterranean environment, presents leaves covered by an extensive indumentum, and a mesophyll full of solid inclusions. The morphology and ultrastructure of the trichomes, calcium carbonate cystoliths and silicified structures of leaves of F. carica cv Dottato were investigated with light, confocal, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. At the same time, histochemical reactions were also employed to analyse the indumentum composed by glandular and non-glandular trichomes by applying chemical reagents and fluorescence microscopy. Non-glandular and glandular trichomes, capitate, are described. Non-glandular trichomes are unicellular simple, spine-like and present different morphology and sizes. The capitate glandular trichomes are present on leaf adaxial and abaxial surface and consist of one-celled stalk and 3/4 cells spherical head. Histochemical characterisation of leaf hairs revealed the presence of flavonoids, while glandular trichome head cells showed a complex mixture of alkaloids, essential oil and flavonoids. Cu and Al were found in the constitutive structures, spike and dome, of the cystoliths. Several epidermal cells and non-glandular trichomes were silicified. Leaf hairs, trichomes secretions, solid inclusions and silicification of F. carica leaf have significant roles to play in relation to leaf protection from external factors, including high-intensity radiation, herbivores or pathogens.     

Heterogeneity of silica and glycan-epitope distribution in epidermal idioblast cell walls in Adiantum raddianum laminae

Laminae of Adiantum raddianum Presl., a fern belonging to the family Pteridaceae, are characterised by the presence of epidermal fibre-like cells under the vascular bundles. These cells were thought to contain silica bodies, but their thickened walls leave no space for intracellular silica suggesting it may actually be deposited within their walls. Using advanced electron microscopy in conjunction with energy dispersive X-ray microanalysis we showed the presence of silica in the cell walls of the fibre-like idioblasts. However, it was specifically localised to the outer layers of the periclinal wall facing the leaf surface, with the thick secondary wall being devoid of silica. Immunocytochemical experiments were performed to ascertain the respective localisation of silica deposition and glycan polymers. Epitopes characteristic for pectic homogalacturonan and the hemicelluloses xyloglucan and mannan were detected in most epidermal walls, including the silica-rich cell wall layers. The monoclonal antibody, LM6, raised against pectic arabinan, labelled the silica-rich primary wall of the epidermal fibre-like cells and the guard cell walls, which were also shown to contain silica. We hypothesise that the silicified outer wall layers of the epidermal fibre-like cells support the lamina during cell expansion prior to secondary wall formation. This implies that silicification does not impede cell elongation. Although our results suggest that pectic arabinan may be implicated in silica deposition, further detailed analyses are needed to confirm this. The combinatorial approach presented here, which allows correlative screening and in situ localisation of silicon and cell wall polysaccharide distribution, shows great potential for future studies.     

Histological Observations on Initiation and Morphogenesis in Immature and Mature Embryo Derived Callus of Barley (Hordeum vulgare L.)

Callus was induced from immature and mature embryos of barley(cv. Haruna Nijo) on Murashige and Skoog medium containing 2mg l^-1 2,4-D and 5 mg l^-1 picloram, respectively. Paraffin sections(10 µm thick) were prepared for histology during callusinitiation and plant regeneration. Meristems were regeneratedfrom nodular compact callus (NC) derived from scutellar epidermisin immature embryos, whereas they were regenerated from NC derivedfrom epidermal cells of leaf or coleoptile bases in mature embryos.Regardless of the explant source, regeneration was predominantlythrough organogenesis, although regeneration through somaticembryogenesis infrequently occurred. Thus, the callus inducedfrom immature and mature embryos of barley was regarded as 'nodularcompact' rather than 'embryogenic'.Copyright 1995, 1999 AcademicPress Barley, callus, Hordeum vulgare, histology, immature embryo, mature embryo, regeneration