Silica deposition in abaxial epidermis before the opening of leaf blades of Pleioblastus chino (Poaceae, Bambusoideae)

BACKGROUND AND AIMS Silica deposition is one of the important characteristics of the family Poaceae. The distribution, deposition process and physiology of silica in this family have been extensively investigated. Bamboos among members of Poaceae have leaves with a fairly long life span, and the leaves continuously accumulate silica in their tissues throughout their life, not only during the course of leaf opening, but also after opening. It has been revealed that the silica deposition process in relation to ageing of the bamboo leaf after opening differed depending on the cell types comprising the tissues. However, silica deposition has never been examined during the development and maturation periods of bamboo leaves. Hence, to clarify the silica deposition process in a developmental stage of the bamboo leaf, distribution of silica was observed in the abaxial epidermis before the opening of the leaf blades of Pleioblastus chino. METHODS: Abaxial epidermal tissues of leaves were examined using a scanning electron microscope equipped with an energy dispersive X-ray microanalyser. KEY RESULTS: Among seven cell types comprising the abaxial epidermis, three types of cells, guard cells, prickle hairs and silica cells, deposited silica conspicuously, and another four types, cork cells, long cells, micro hairs and subsidiary cells, deposited only a little silica. Among the former group of cell types, silica cells and guard cells deposited silica over their entire surfaces, while prickle hairs deposited silica only in the point-tips. Silica deposition was detected firstly in prickle hairs, and then in silica cells and guard cells. Only silica cells were assumed to deposit silica conspicuously before leaf opening but not conspicuously after opening. CONCLUSIONS: Cell types in leaf epidermis of bamboo are classified into three groups according to the silica deposition pattern. Silica deposition in silica cells may be positive as a part of the physiological activities of leaves.     

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     

Silicon distribution and accumulation in shoot tissue of the tropical forage grass Brachiaria brizantha

Silicon (Si) accumulation in organs and cells is one of the most prominent characteristics of plants of the family Poaceae. Many species from this family are used as forage plants for animal feeding. The present study investigates in Brachiaria brizantha (Hochst. ex A. Rich.) Stapf. cv. Marandu: (1) the dry matter production and Si content in shoot due to soil Si fertilizations; (2) the Si distribution among shoot parts; and (3) the silica deposition and localization in leaves. Plants of B. brizantha cv. Marandu were grown under contrasting Si supplies in soil and nutrient solution. Silica deposition and distribution in grass leaf blades were observed using light microscope and scanning electron microscope equipped with an energy dispersive X-ray spectrometer (SEM-EDXS). Silicon concentration in the B. brizantha shoot increased according to the Si supply. Silicon in grass leaves decreased following the order: mature leaf blades > recently expanded leaf blades > non-expanded leaf blades. Silicon accumulates mainly on the upper (adaxial) epidermis of the grass leaf blades and, especially, on the bulliform cells. The Si distribution on adaxial leaf blade surface is non uniform and reflects a silica deposition exclusively on the cell wall of bulliform cells.     

Do lignification and silicification of the cell wall precede silicon deposition in the silica cell of the rice (Oryza sativa L.) leaf epidermis?

Aims

Rice is a well-known silica-accumulating plant. The dumbbell-shaped silica bodies in the silica cells in rice leaf epidermis are formed via biosilicification, but the underlying mechanisms are largely unknown.

Methods

Leaves at different developmental stages were collected to investigate silica cell differentiation by analyzing structures and silicon localization in the silica cells.

Results

Exogenous silicon application increased both shoot and root biomass. When silicon was supplied, silica cells in the leaf epidermis developed gradually into a dumbbell-shape and became increasingly silicified as leaves aged. Silicon deposition in the silica cells was not completed until the leaf was fully expanded. Multiple lines of evidence suggest that lignification of silica cell walls precedes silicon deposition in the lumen of silica cells. The organized needle-like silica microstructures were formed by moulding the inner cell walls and filling up the lumen of the silica cell following leaf maturation.

Conclusions

Two processes were involved in silicon deposition: (1) the silica cell wall was lignified and silicified, and then (2) the silicon was deposited gradually in silica cells as leaves aged. Silica body formation was not completed until the leaf was fully mature.     

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.     

Scanning electron microscopy of silica bodies and other epidermal features in Gibasis (Tradescantia) leaf

The scanning electron microscope is used to examine epidermal preparations belonging to three species of Gibasis , a genus allied to Tradescantia. The surface topography of the leaf is exposed for investigation at high magnifications, the observations add new information about structural projections from the surface and contribute to an understanding of silica deposition and its organization. Hairs of three main types occur: (1) short two-celled trichomes, hook or prickle hairs, (2) long uniseriate hairs consisting of four to six cells, (3) three-celled glandular micro-hairs. The arrangement and shape of epidermal cells and stomata are reported. Epidermal papillae were observed for the first time in two of the three species; their morphology is described in detail and their spacing expressed mathematically. Variations in the structure and distribution of silica cells are depicted, and new aspects of the silica bodies are displayed in relief. The taxonomic significance of these anatomical characters is discussed in relation to species and chromosome numbers.     

Salt excretion through the cuticle without disintegration of fine structures in the salt glands of Rhodes grass (Chloris gayana Kunth)

Some salt-tolerant plants belonging to the Poaceae possess bicellular salt glands that excrete salt-containing water. To clarify the excretion process from the outer cell of the bicellular salt gland (cap cell), unwashed and washed fresh leaves of Rhodes grass (Chloris gayana Kunth), Poaceae, were cryo-fixed rapidly, and the surface fine structures of the leaves were observed by cryo-scanning electron microscopy with high resolution. The cuticle on the cap cell did not have any pores or signs of rupturing. The cuticle of the cap cells lacks the epicuticular waxes that cover most surfaces of leaf epidermis. After excreted droplets on the salt glands were completely removed by washing with water, re-excreted droplets were observed 3 h later. These findings indicate that the bicellular salt glands of Rhodes grass excrete salt-containing water continuously through the wax-free cuticle of the cap cell without disintegration of the cuticular structure.     

Anatomical features and ultrastructure of Deschampsia antarctica (Poaceae) leaves from different growing habitats

BACKGROUND AND AIMS: The leaf anatomy and ultrastructure of Deschampsia antarctica (Poaceae) plants growing in three different habitats (a dry site in the Antarctic tundra, a wet site in a zone exposed to sea spray and a greenhouse) were investigated. The ultrastructure of the leaves of D. antarctica has not been studied before. METHODS: Semi-thin sections of the D. antarctica leaves were stained with toluidine blue and viewed using a light microscope. Ultra-thin sections stained with uranyl acetate and lead citrate were examined using a transmission electron microscope. KEY RESULTS: Plants growing in the Antarctic tundra and in a greenhouse had stronger xerophytic features than those growing at the seashore. The stress response of D. antarctica plants growing in the wet environment, exposed to high salinity and flooding, included: irregular mesophyll cells, large intercellular spaces in the parenchymatic layer, bulliform epidermal cells and vascular bundles surrounded with deformed outer and inner bundle sheaths of leaves. The highest number of sclerenchymatic fibres is characteristic of the leaves of plants growing in a greenhouse, whereas the smallest was of plants growing in a wet habitat. Stress conditions can disturb the formation of sclerenchymatic fibres. In plants growing in the Maritime Antarctic the chloroplasts of the mesophyll cells of leaves are of an irregular shape, with pockets or invaginations inside the organelles and outgrowths. Both of them make the surfaces of chloroplasts larger, and result in an increase in the amount of substances exchanged between the chloroplasts and cytoplasm or the other organelles. The leaf mesophyll cells of D. antarctica plants growing in Antarctica contain atypical structures including numerous vesicles of different sizes and concentrically arranged membranes. CONCLUSIONS: The anatomical and ultrastructural features of the leaf and their changes under stress conditions are considered in relation to the adaptations of D. antarctica to the climate conditions in the Maritime Antarctic.     

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.     

Silicification in sorghum (Sorghum bicolor) cultivars with different drought tolerance

Sorghum belongs to a group of economically important, silicon accumulating plants. X-ray microanalysis coupled with environmental scanning electron microscopy (ESEM) of fresh root endodermal and leaf epidermal samples confirms histological and cultivar specificity of silicification. In sorghum roots, silicon is accumulated mostly in endodermal cells. Specialized silica aggregates are formed predominantly in a single row in the form of wall outgrowths on the inner tangential endodermal walls. The density of silica aggregates per square mm of inner tangential endodermal cell wall is around 2700 and there is no significant difference in the cultivars with different content of silicon in roots. In the leaf epidermis, silicon deposits were present in the outer walls of all cells, with the highest concentration in specialized idioblasts termed 'silica cells'. These cells are dumb-bell shaped in sorghum. In both the root endodermis and leaf epidermis, silicification was higher in a drought tolerant cultivar Gadambalia compared with drought sensitive cultivar Tabat. Silicon content per dry mass was higher in leaves than in roots in both cultivars. The values for cv. Gadambalia in roots and leaves are 3.5 and 4.1% Si, respectively, and for cv. Tabat 2.2 and 3.3%. However, based on X-ray microanalysis the amount of Si deposited in endodermal cell walls in drought tolerant cultivar (unlike the drought susceptible cultivar) is higher than that deposited in the leaf epidermis. The high root endodermal silicification might be related to a higher drought resistance.     

Silicification of bamboo (Phyllostachys heterocycla Mitf.) root and leaf

Bamboo is a silicon accumulating plant. In leaves, the major place of silicon (Si) deposition is the epidermis, with the highest concentration of Si in silica cells. In bamboo roots, the deposition of Si is found only in endodermal cell walls. The silicification of leaves and roots was examined in the economically important bamboo plant Phyllostachys heterocycla, using an environmental scanning electron microscope coupled with X-ray microanalysis, as well as gravimetric quantification. The content of Si on a dry weight basis measured by gravimetric quantification was 7.6% in leaves and 2.4% in roots, respectively. Moreover, quantification of EDX data showed high Si impregnation of the inner tangential endodermal walls. Si content in this part of the root endodermal cell walls was even higher than that in the outer leaf epidermal walls, where conspicuous deposition of Si often occurs in grass plants.     

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.     

Relationships between photosynthetic activity and silica accumulation with ages of leaf in Sasa veitchii (Poaceae, Bambusoideae)

BACKGROUND AND AIMS: Bamboos have long-lived, evergreen leaves that continue to accumulate silica throughout their life. Silica accumulation has been suggested to suppress their photosynthetic activity. However, nitrogen content per unit leaf area (N(area)), an important determinant of maximum photosynthetic capacity per unit leaf area (P(max)), decreases as leaves age and senescence. In many species, P(max) decreases in parallel with the leaf nitrogen content. It is hypothesized that if silica accumulation affects photosynthesis, then P(max) would decrease faster than N(area), leading to a decrease in photosynthetic rate per unit leaf nitrogen (photosynthetic nitrogen use efficiency, PNUE) with increasing silica content in leaves. METHODS: The hypothesis was tested in leaves of Sasa veitchii, which have a life span of 2 years and accumulate silica up to 41 % of dry mass. Seasonal changes in P(max), stomatal conductance, N(area) and silica content were measured for leaves of different ages. KEY RESULTS: Although P(max) and PNUE were negatively related with silica content across leaves of different ages, the relationship between PNUE and silica differed depending on leaf age. In second-year leaves, PNUE was almost constant although there was a large increase in silica content, suggesting that leaf nitrogen was a primary factor determining the variation in P(max) and that silica accumulation did not affect photosynthesis. PNUE was strongly and negatively correlated with silica content in third-year leaves, suggesting that silica accumulation affected photosynthesis of older leaves. CONCLUSIONS: Silica accumulation in long-lived leaves of bamboo did not affect photosynthesis when the silica concentration of a leaf was less than 25 % of dry mass. Silica may be actively transported to epidermal cells rather than chlorenchyma cells, avoiding inhibition of CO2 diffusion from the intercellular space to chloroplasts. However, in older leaves with a larger silica content, silica was also deposited in chlorenchyma cells, which may relate to the decrease in PNUE.     

Detection of Biogenic Silica in Leaf Blade, Leaf Sheath, and Stem of Bermuda Grass (Cynodon dactylon) Using LIBS and Phytolith Analysis

Laser induced breakdown spectroscopy (LIBS) has been used to perform in situ analysis of major and minor elements present in the different parts of the Bermuda grass (Cynodon dactylon). In situ, point detection/analysis of the elements in plants without any sample preparation has been demonstrated. LIBS spectra of the different parts (leaf blade, leaf sheath and stem) of fresh C. dactylon plant have been recorded to study the pattern of silica deposition in its different parts. Atomic lines of Si, Mg, Ca, C, Al, Zn, N, Sr, etc. have been observed in the LIBS spectra of the C. dactylon. A close observation of LIBS spectra of the different parts of the plants shows that silica concentration is greater in leaf blades than leaf sheaths and stems. The results obtained with LIBS analysis are also compared with the number density of phytoliths deposited in different parts of C. dactylon. It is observed that the highest silicified cell frequency is present in leaf blades followed by leaf sheaths and stems which is in close agreement with LIBS analysis.     

中国菊科绢蒿属植物叶表皮特征观察

利用扫描电镜对17种绢蒿属植物叶表皮特征进行了观察。结果表明,该属植物叶表面有大量带状蜡质和表皮毛覆盖,在蜡质成分的外面分泌有盐分颗粒,表现出明显的旱生结构特征。表皮毛在种间无明显差异,而气孔器在保卫细胞形状和表面纹饰等特征组合方面存在较大差异,说明叶表皮气孔器特征对于绢蒿属植物的系统分类有参考价值。     

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.     

Micromorphology of the leaf epidermis in taxa of the Agropyron-Elymus complex (Poaceae)

WEBB, M. E. & ALMEIDA, M. T., 1990. Micromorphology of the leaf epidermis in taxa of the Agropyron-Elymus complex (Poaceae). A comparative analysis by scanning electron microscopy was carried out on both leaf epidermes (adaxial and abaxial) of Elymus pychnanthus (Godr.) Meld, and Agropyron glaucum Roemer & Schultes.
The adaxial epidermes of E.pychnanthus and A. glaucum are similar in the position and the shape of the long cells, silica bodies and costal and papillate prickles. They differ in the higher number of silica cells in E. pychnanthus and in the presence of intercostal hooks in A. glaucum.
The abaxial epidermes of both species are similar in the shape of interstomatal cells, silica bodies and papillate prickles. They differ in the distribution of the stomata and in the contact zones of the long cells. Elymus pychnanthus and A. glaucum differ also in the trichomes situated along the margins of the leaf blade.
These micromorphological differences, especially those of the abaxial epidermis, are useful taxonomic features.     

Nitrogen Use within the Growing Leaf Blade of Tall Fescue

Leaf elongation rate (LER) of grasses depends on N supply and is expressed mostly through cell production, whereas most N in mature leaf tissues is chloroplastic. Our objective was to evaluate a possible competition for N between cell production and chloroplast development processes, utilizing the gradient of cell development along the leaf growth zone of tall fescue (Festuca arundinacea Schreb.). Under the two contrasting N regimes, total N content was highest in the cell production zone, declined sharply as cells elongated, and remained relatively constant in more distal positions, at values close to those measured in mature tissues. A similar pattern was found for N in proteins and nucleic acids that were not soluble in 80% ethanol. Content of N compounds soluble in 80% ethanol was higher in the cell production and elongation zones than in mature parts of the leaf. NO3- N content was low in the cell production zone and increased in the cell elongation zone for high-N plants. The deposition rate of total N in the growth zone was much higher with plants in high N than in those shifted to no N. For both N regimes, most N was deposited during cell production and early cell elongation. Little N was deposited during cell maturation where ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) was being actively synthesized. This suggests that synthesis of Rubisco, and probably other chloroplastic proteins, occurs largely from recycling of N that was previously incorporated into proteins during cell production. Thus, Rubisco content in mature tissues is more closely associated with N deposited during cell production than with N deposited during its biosynthesis.