Calcium carbonate deposition in a cell wall sac formed in mulberry idioblasts

Summary. Although calcium carbonate is known to be a common biomineral in plants, very little attention has been given to the biological control of calcium carbonate deposition. In mulberry leaves, a subcellular structure is involved in mineral deposition and is described here by a variety of cytological techniques. Calcium carbonate was deposited in large, rounded idioblast cells located in the upper epidermal layer of mulberry leaves. Next to the outmost region (“cap”) of young idioblasts, we found that the inner cell wall layer expanded to form a peculiar outgrowth, named cell wall sac in this report. This sac grew and eventually occupied the entire apoplastic space of the idioblast. Inside the mature cell wall sac, various cellulosic membranes developed and became the major site of Ca carbonate deposition. Concentrated Ca²⁺ was pooled in the peripheral zone, where small Ca carbonate globules were present in large numbers. Large globules were tightly packed among multiple membranes in the central zone, especially in compartments formed by cellulosic membranes and in their neighboring membranes. The maximum Ca sink capacity of a single cell wall sac was quantified using enzymatically isolated idioblasts as approximately 48 ng. The newly formed outgrowth in idioblasts is not a pure calcareous body but a complex cell wall structure filled with substantial amounts of Ca carbonate crystals. Correspondence and reprints: Graduate School of Science and Technology, Kyoto Institute of Technology, Goshokaido, Matsugasaki, Sakyo, Kyoto 606-8585, Japan.     

Calcium channels are involved in calcium oxalate crystal formation in specialized cells of Pistia stratiotes L

BACKGROUND AND AIMS: Pistia stratiotes produces large amounts of calcium (Ca) oxalate crystals in specialized cells called crystal idioblasts. The potential involvement of Ca(2+) channels in Ca oxalate crystal formation by crystal idioblasts was investigated. METHODS: Anatomical, ultrastructural and physiological analyses were used on plants, fresh or fixed tissues, or protoplasts. Ca(2+) uptake by protoplasts was measured with (45)Ca(2+), and the effect of Ca(2+) channel blockers studied in intact plants. Labelled Ca(2+) channel blockers and a channel protein antibody were used to determine if Ca(2+) channels were associated with crystal idioblasts. KEY RESULTS: (45)Ca(2+) uptake was more than two orders of magnitude greater for crystal idioblast protoplasts than mesophyll protoplasts, and idioblast number increased when medium Ca was increased. Plants grown on media containing 1-50 microM of the Ca(2+) channel blockers, isradipine, nifedipine or fluspirilene, showed almost complete inhibition of crystal formation. When fresh tissue sections were treated with the fluorescent dihydropyridine-type Ca(2+) channel blocker, DM-Bodipy-DHP, crystal idioblasts were intensely labelled compared with surrounding mesophyll, and the label appeared to be associated with the plasma membrane and the endoplasmic reticulum, which is shown to be abundant in idioblasts. An antibody to a mammalian Ca(2+) channel alpha1 subunit recognized a single band in a microsomal protein fraction but not soluble protein fraction on western blots, and it selectively and heavily labelled developing crystal idioblasts in tissue sections. CONCLUSIONS: The results demonstrate that Ca oxalate crystal idioblasts are enriched, relative to mesophyll cells, in dihydropyridine-type Ca(2+) channels and that the activity of these channels is important to transport and accumulation of Ca(2+) required for crystal formation.     

Subcellular compartmentation of strontium and zinc in mulberry idioblasts in relation to phytoremediation potential

The cellular compartmentation of heavy metals was analyzed using mulberry leaves, in which CaCO₃-forming idioblasts are situated in the epidermal layer. Germinated mulberry seedlings were grown on hydroponic culture medium containing strontium (Sr), zinc (Zn), and cadmium (Cd) with and without supplemental calcium (Ca). After ten weeks of growth, toxic effects of these metals were assessed by measuring shoot length and chlorophyll content of leaves. Sr and Cd treatment at a higher dose (4 mM for Sr, 25 μM for Cd) resulted in signs of toxicity, whereas no distinct phytotoxicity was observed at 500 μM Zn. Elemental mapping of leaves using an energy-dispersive X-ray microanalysis system fitted to variable-pressure scanning electron microscope showed that Sr and Zn were preferentially accumulated in the idioblasts and Cd was not detected in any type of leaf cell. The deposition site of Sr was confined to cell wall sacs developing in idioblasts. The Sr sink capacity in leaves was more than 30 mg/g dry weight, which equaled the Ca sink capacity. Exposure of Sr + Ca led to the co-localization of Sr and Ca in the same cell wall sac, in which Ca and Sr deposition were each estimated to be 60–80 ng. The localization site of Zn was cell walls of a dome-shaped protrusion (cap) of idioblasts, together with silicon (Si) originating as a contaminant in tap water used for culturing. Mulberry idioblasts were unique in showing metal-dependent distribution to two subcellular sites (cell wall sac and cap region) of idioblasts. In conclusion, mulberry plant is a candidate for phytoremediation of radiostrontium because of their Sr-hyperaccumulating capacity with sufficient leaf biomass production.     

The Role of Druse and Raphide Calcium Oxalate Crystals in Tissue Calcium Regulation in Pistia stratiotes Leaves

Abstract: Ca oxalate crystal formation was examined in Pistia stratiotes L. leaves during excess Ca and Ca-deficient conditions. Pistia produces druse crystal idioblasts in the adaxial mesophyll and raphide idioblasts in the abaxial aerenchyma. Raphide crystals were previously found to grow bidirectionally, and here we show that Ca is incorporated along the entire surfaces of developing druse crystals, which are coated with membrane-bound microprojections. Leaves formed on plants grown on 0 Ca medium have fewer and smaller druse crystals than leaves formed under 5 mM Ca ("control") conditions, while raphide crystal formation is completely inhibited. When plants were moved from 0 to 15 mM ("high") Ca, the size and number of crystals in new leaves returned to (druse) or exceeded (raphide) control levels. High Ca also induced formation of druse, but not raphide, crystals in differentiating chlorenchyma cells. When plants were transferred from 15 mM Ca to 0 Ca, young druse crystals were preferentially partially dissolved. Oxalate oxidase, an enzyme that degrades oxalate, increased during Ca deficiency and was localized to the crystal surfaces. The more dynamic nature of druse crystals is not due to hydration form as both crystal types are shown to be monohydrate. Part of the difference may be because raphide idioblasts have developmental constraints that interfere with a more flexible response to changing Ca. These studies demonstrate that excess Ca can be stored as Ca oxalate, the Ca can be remobilized under certain conditions, and different forms of Ca oxalate have different roles in bulk Ca regulation.     

Organ-Level Analysis of Idioblast Patterning in Egeria densa Planch. Leaves

Leaf tissues of plants usually contain several types of idioblasts, defined as specialized cells whose shape and contents differ from the surrounding homogeneous cells. The spatial patterning of idioblasts, particularly of trichomes and guard cells, across the leaf epidermis has received considerable attention as it offers a useful biological model for studying the intercellular regulation of cell fate and patterning. Excretory idioblasts in the leaves of the aquatic monocotyledonous plant Egeria densa produced light blue autofluorescence when irradiated with ultraviolet light. The use of epifluorescence microscopy to detect this autofluorescence provided a simple and convenient method for detecting excretory idioblasts and allowed tracking of those cells across the leaf surfaces, enabling quantitative measurement of the clustering and spacing patterns of idioblasts at the whole leaf level. Occurrence of idioblasts was coordinated along the proximal–distal, medial–lateral, and adaxial–abaxial axes, producing a recognizable consensus spatial pattern of idioblast formation among fully expanded leaves. Idioblast clusters, which comprised up to nine cells aligned along the proximal–distal axis, showed no positional bias or regularity in idioblast-forming areas when compared with singlet idioblasts. Up to 75% of idioblasts existed as clusters on every leaf side examined. The idioblast-forming areas varied between leaves, implying phenotypic plasticity. Furthermore, in young expanding leaves, autofluorescence was occasionally detected in a single giant vesicle or else in one or more small vesicles, which eventually grew to occupy a large portion of the idioblast volume as a central vacuole. Differentiation of vacuoles by accumulating the fluorescence substance might be an integral part of idioblast differentiation. Red autofluorescence from chloroplasts was not detected in idioblasts of young expanding leaves, suggesting idioblast differentiation involves an arrest in chloroplast development at a very early stage, rather than transdifferentiation of chloroplast-containing epidermal cells.     

Cellular ultrastructure and crystal development in Amorphophallus (Araceae)

Background and Aims: Species of Araceae accumulate calcium oxalate in the form ofcharacteristically grooved needle-shaped raphide crystals andmulti-crystal druses. This study focuses on the distributionand development of raphides and druses during leaf growth inten species of Amorphophallus (Araceae) in order to determinethe crystal macropatterns and the underlying ultrastructuralfeatures associated with formation of the unusual raphide groove. Methods: Transmission electron microscopy (TEM), scanning electron microscopy(SEM) and both bright-field and polarized-light microscopy wereused to study a range of developmental stages. Key Results: Raphide crystals are initiated very early in plant development.They are consistently present in most species and have a fairlyuniform distribution within mature tissues. Individual raphidesmay be formed by calcium oxalate deposition within individualcrystal chambers in the vacuole of an idioblast. Druse crystalsform later in the true leaves, and are absent from some species.Distribution of druses within leaves is more variable. Drusesinitially develop at leaf tips and then increase basipetallyas the leaf ages. Druse development may also be initiated incrystal chambers. Conclusions: The unusual grooved raphides in Amorphophallus species probablyresult from an unusual crystal chamber morphology. There aremultiple systems of transport and biomineralization of calciuminto the vacuole of the idioblast. Differences between raphideand druse idioblasts indicate different levels of cellular regulation.The relatively early development of raphides provides a defensivefunction in soft, growing tissues, and restricts build-up ofdangerously high levels of calcium in tissues that lack theability to adequately regulate calcium. The later developmentof druses could be primarily for calcium sequestration.     

ULTRASTRUCTURE OF DRUSE CRYSTAL IDIOBLASTS IN LEAVES OF CERCIDIUM FLORIDUM

The ultrastructure of druse crystal idioblasts in palo verde leaves is similar in certain aspects to other crystal-containing idioblasts, but also displays several notable differences. Although the crystal itself is dissolved during the preparative procedure, the druse idioblast is readily observable. The large crystal is contained in a tightly appressed vacuole in the center of the idioblast. Between the crystal vacuole and the cell wall there is a narrow stalk-like connection which has the same substructure and staining characteristics as cell wall material. The membrane of this crystal vacuole and the idioblast plasmalemma stain asymmetrically, while other cellular membranes in the idioblast appear symmetrical. Much of the remaining cell volume is occupied by a ramified vacuome and a peripherally displaced nucleus. The plastids of the druse idioblast are markedly different from chloroplasts in adjacent parenchyma cells. The former lack the size, starch grains, and well-developed grana of the latter. Idioblast mitochondria are similar in quantity and appearance to those of palisade cells, except for a greater number of cristae in the former. Dictyosomes, while rare in mesophyll cells, are quite common in the idioblast. These features suggest that the druse crystal idioblast is metabolically active and not dead at maturity.     

Differential targeting of the tobacco PR-1 pathogenesis-related proteins to the extracellular space and vacuoles of crystal idioblasts.

Several biochemical and localization studies have shown that the acidic isoforms of the tobacco pathogenesis-related (PR) proteins, PR-1a, -1b and -1c are secreted to the extracellular spaces of leaves in response to pathogen infection or chemical treatment. Here we report the differential accumulation of these proteins within the vacuoles of specialized cells known as crystal idioblasts. In situ hybridization analysis indicated that crystal idioblasts expressed the PR-1 genes at the mRNA level and suggested that PR-1 proteins were synthesized by these cells. Transgenic plants which constitutively express a chimeric gene encoding an acidic PR-1b isoform also accumulated PR-1 protein in the extracellular spaces and within crystal idioblast vacuoles. Analysis of mRNA derived from these transgenic plants indicated that expression of the introduced PR-1b gene was responsible for the accumulation of PR-1 protein in these two distinct locations. The synthesis and accumulation within crystal idioblasts of PR-1 proteins, which are secreted by other cell types, indicates that idioblasts sort these proteins in a unique manner. Moreover, this suggests that protein sorting in higher plants may be modulated in a cell specific manner.     

Cell and calcium oxalate crystal growth is coordinated to achieve high-capacity calcium regulation in plants

Summary Crystal idioblasts are cells which are specialized for accumulation of Ca²⁺ as a physiologically inactive, crystalline salt of oxalic acid. Using microautoradiographic, immunological, and ultrastructural techniques, the process of raphide crystal growth, and how crystal growth is coordinated with cell growth, was studied in idioblasts ofPistia stratiotes. Incorporation of⁴⁵Ca²⁺ directly demonstrated that, relative to surrounding mesophyll cells, crystal idioblasts act as high-capacity Ca²⁺ sinks, accumulating large amounts of Ca²⁺ within the vacuole as crystals. The pattern of addition of Ca²⁺ during crystal growth indicates a highly regulated process with bidirectional crystal growth. In very young idioblasts,⁴⁵Ca²⁺ is incorporated along the entire length of the needle-shaped raphide crystals, but as they mature incorporation only occurs at crystal tips in a bidirectional mode. At full maturity, the idioblast stops Ca²⁺ uptake, although the cells are still alive, demonstrating an ability to strictly regulate Ca transport processes at the plasma membrane. In situ hybridization for ribosomal RNA shows young idioblasts are extremely active cells, are more active than older idioblasts, and have higher general activity than surrounding mesophyll cells. Polarizing and scanning electron microscopy demonstrate that the crystal morphology changes as crystals develop and includes morphological polarity and an apparent nucleation point from which crystals grow bidirectionally. These results indicate a carefully regulated process of biomineralization in the vacuole. Finally, we show that the cytoskeleton is important in controlling the idioblast cell shape, but the regulation of crystal growth and morphology is under a different control mechanism.Abbreviation SEM scanning electron microscopy     

Biosynthesis of L-ascorbic acid and conversion of carbons 1 and 2 of L-ascorbic acid to oxalic acid occurs within individual calcium oxalate crystal idioblasts

L-Ascorbic acid (AsA) and its metabolic precursors give rise to oxalic acid (OxA) found in calcium oxalate crystals in specialized crystal idioblast cells in plants; however, it is not known if AsA and OxA are synthesized within the crystal idioblast cell or transported in from surrounding mesophyll cells. Isolated developing crystal idioblasts from Pistia stratiotes were used to study the pathway of OxA biosynthesis and to determine if idioblasts contain the entire path and are essentially independent in OxA synthesis. Idioblasts were supplied with various (14)C-labeled compounds and examined by micro-autoradiography for incorporation of (14)C into calcium oxalate crystals. [(14)C]OxA gave heavy labeling of crystals, indicating the isolated idioblasts are functional in crystal formation. Incubation with [1-(14)C]AsA also gave heavy labeling of crystals, whereas [6-(14)C]AsA gave no labeling. Labeled precursors of AsA (L-[1-(14)C]galactose; D-[1-(14)C]mannose) also resulted in crystal labeling, as did the ascorbic acid analog, D-[1-(14)C]erythorbic acid. Intensity of labeling of isolated idioblasts followed the pattern OxA > AsA (erythorbic acid) > L-galactose > D-mannose. Our results demonstrate that P. stratiotes crystal idioblasts synthesize the OxA used for crystal formation, the OxA is derived from the number 1 and 2 carbons of AsA, and the proposed pathway of ascorbic acid synthesis via D-mannose and L-galactose is operational in individual P. stratiotes crystal idioblasts. These results are discussed with respect to fine control of calcium oxalate precipitation and the concept of crystal idioblasts as independent physiological compartments.     

Silicon-accumulating idioblasts in leaves of Cecropiaceae (Urticales)

A survey of the structure and mineral composition of leaf idioblasts in the Cecropiaceae was conducted. In all six genera of the family, idioblasts usually occur as trichomes or enlarged epidermal cells and nearly always accumulate Si, in marked contrast to the idioblasts of other members of the Urticales, which mostly possess cystoliths containing abundant Ca and Si. The horizontally elongate, mineralized structures ofPoikilospermum, reported formerly as “cystoliths” also contain mainly Si and little Ca. Six genera of Cecropiaceae share a common character in accumulating abundant Si in idioblasts of the leaf epidermis, while lacking cystoliths composed of abundant Ca and Si.     

Observations on the Distribution of Mineral Elements in the Leaf of Wheat (Triticum aestivum L.), with Particular Reference to Silicon

Wheat (Triticum aestivum L. cv. Wheaton) plants were grown inwater culture or in soil. Basal leaves (B) were harvested after3 weeks from the water culture plants, while flag leaves werecollected from soil-grown material at the time of inflorescenceemergence (E0) and 7 d after emergence (E + 7). Mineral distributionin bulk frozen leaves was investigated using SEM and X-ray microanalysis.The elements detected were silicon, phosphorus, chlorine, sulphur,potassium and calcium. Potassium was present in all cell typesat all harvests, chlorine was almost entirely confined to theadaxial and abaxial epidermi, while sulphur was only rarelydetected in the E0 and E + 7 leaves. Phosphorus was presentat higher levels in the E + 7 leaves than in the B or E0 leaves.At the B harvest calcium was confined to the adaxial epidermalcells, but in the E0 and E + 7 leaves it was present in bothepidermi. Silicon was, initially, mainly detected in the abaxialepidermal cells, but in older (E + 7) leaves it was presentin both epidermi and in some internal tissues. Mineral transportwithin the leaf and ionic environment at silica deposition sitesare discussed. Wheat, Triticum aestivum L., leaf, mineral distribution, X-ray microanalysis, silicon, calcium     

Mature Raphid and Raphid Idioblast Structure in Plants of the Edible Aroid Genera Colocasia, Alocasia, and Xanthosoma

Mature raphid idioblasts examined in this study appear to consistof a bundle of raphid crystals contained within a polysaccharidematrix. The cytoplasm consists of a thin parietal layer withfew discernible organelles. These cells appear to be true idioblasts,and no intercellular connections were observed between thesecells and adjacent parenchyma cells. Dissolution of the middlelamella occurs in Colocasia and to some extent in Xanthosoma.The mechanism of crystal release in the five species studiedis the same, e.g. a swelling of a polysaccharide material whichbreaks the idioblast wall and forces the release of the raphides.In Colocasia this results in a forceful ejection with releaseof a relatively few number of crystals at any one time. Xanthosomaand Alocasia do not exhibit a forceful ejection of their crystals,but a less forceful release of a crystal mass which then disassociatesto free the individual crystals. The raphides differ in structureand size, however, they have three structural features in common.First, the crystals have two distinct points, one tapering toan elongate point, the other abruptly pointed. Second, the crystalshave surface barbs with tips oriented away from the taperingpoint. And third, deep grooves are present along the lengthof the crystals. The possible relationship between raphid structureand the acrid nature of the idioblasts is discussed.     

Calcium Oxalate Raphide Crystals and Crystalliferous Idioblasts in the Carpels of Ornithogalum caudatum

Crystalliferous idioblasts commonly are found in groups of twoor three cells in the peripheral region of the carpels Crystals,composed of calcium oxalate, usually are m well-organized bundleswhich develop within a matrix of protein and carbohydrate inthe vacuole of each idioblast The matrix occurs around and betweenindividual crystal chambers and contains spheres and tubules5.4 nm in diameter The matrix changes in character and locationwith age Crystals form within their own individual chambers,each comprised of a series of lamellae The number of lamellaeis variable The innermost lamella is different from the othersin that it is apparently continuous The other lamellae are platelikeand superficially resemble successive periderms. The lamellaemay begin and/or terminate abruptly or they may anastamose Eachlamella is composed of chains of spheres about 6 1 nm in diameterand is separated from adjacent lamellae by tubules 5.4 nm indiameter Both the crystals and slime body are absorbed duringlater stages of carpel maturation. Ornithogalum caudatum Ait carpel, calcium oxalate, idioblasts, ultrastructure     

Distribution of Assimilates from Various Source Leaves During the Development of Gladiolus grandiflorus

The distribution pattern of the products of photosynthesis wasstudied in gladiolus plants (Gladiolus grandiflorus cv Eurovision)in four stages of development I, plants having a very younginflorescence still enclosed between the leaves; II, plantswith a young inflorescence just emerged from the leaves, III,plants at full bloom, IV, plants with young fruits. The first,third or sixth foliage leaf was labelled with ¹⁴CO₂, and subsequentdistribution in the plant was determined Results were expressedas a percentage of translocated ¹⁴C accumulated by each partof the plant which gives a measure of its ‘sink strength’,or as ‘relative sink activity’ (RSA) which is independentof the size of the indicated organ. There are two competing sinks in the developing gladiolus—theinflorescence and the new corm. When RSA is the criterton theinflorescence constitutes the main sink irrespective of thesource leaf from the first stage until flowering. With the subsequentwilting of the flowers and fruit set RSA of the inflorescencedeclines rapidly and the new corm becomes the main sink When‘sink strength’ is the criterton it appears thatthe inflorescence acts as a very weak sink when it is youngand becomes increasingly stronger until flowering and then declinessteeply. Sink strength of the corm declines during the developmentof the inflorescence and then increases again steeply with wiltingof the flowers and fruit set. There are small differences betweenthe various source leaves. The young new corm acts as a strongsink for the lower foliage leaf and progressively weaker forupper leaves. Gladiolus grandiflorus, flower development, corm, assimilates distribution, sink strength, relative sink activity     

Difference in light-induced increase in ploidy level and cell size between adaxial and abaxial epidermal pavement cells of Phaseolus vulgaris primary leaves

Changes in nuclear DNA content and cell size of adaxial andabaxial epidermal pavement cells were investigated using brightlight-induced leaf expansion of Phaseolus vulgaris plants. Inprimary leaves of bean plants grown under high (sunlight) ormoderate (ML; photon flux density, 163 µmol m^–2s^–1) light, most adaxial epidermal pavement cells hada nucleus with the 4C amount of DNA, whereas most abaxial pavementcells had a 2C nucleus. In contrast, plants grown under lowintensity white light (LL; 15 µmol m^–2 s^–1)for 13 d, when cell proliferation of epidermal pavement cellshad already finished, had a 2C nuclear DNA content in most adaxialpavement cells. When these LL-grown plants were transferredto ML, the increase in irradiance raised the frequency of 4Cnuclei in adaxial but not in abaxial pavement cells within 4d. On the other hand, the size of abaxial pavement cells increasedby 53% within 4 d of transfer to ML and remained unchanged thereafter,whereas adaxial pavement cells continuously enlarged for 12d. This suggests that the increase in adaxial cell size after4 d is supported by the nuclear DNA doubling. The differentresponses between adaxial and abaxial epidermal cells were notinduced by the different light intensity at both surfaces. Itwas shown that adaxial epidermal cells have a different propertythan abaxial ones. Key words: Cell enlargement, endopolyploidization, epidermal pavement cells, incident light intensity, leaf expansion, nuclear DNA content, Phaseolus vulgaris     

L-Ascorbic acid and L-galactose are sources for oxalic acid and calcium oxalate in Pistia stratiotes

Axenic Pistia stratiotes L. plants were pulse-chase labeled with [14C]oxalic acid, L[1-14C]ascorbic acid, L-6-14C]ascorbic acid, D-[1-14C]erythorbic acid, L-[1-14C]galactose, or [1-14C]glycolate. Specific radioactivities of L-ascorbic acid (AsA), free oxalic acid (OxA) and calcium oxalate (CaOx) in labeled plants were compared. Samples of leaf tissue were fixed for microautoradiography and examined by confocal microscopy. Results demonstrate a biosynthetic role for AsA as precursor of OxA and its crystalline deposition product, CaOx, in idioblast cells of P. stratiotes and support the recent discovery of Wheeler, Jones and Smirnoff (Wheeler, G.L., Jones M.A., & Smirnoff, N. (1998). The biosynthetic pathway of vitamin C in higher plants. Nature, 393, 365-369) that L-galactose is a key intermediate in the conversion of D-glucose to AsA in plants. D-[1-14C]erythorbic acid (a diastereomeric analog of AsA) is utilized also by P. stratiotes as a precursor of OxA and its calcium salt deposition product in idioblasts. Labeled OxA is rapidly incorporated into CaOx in idioblasts, but microautoradiography shows there is also significant incorporation of carbon from OxA into other components of growing cells, contrary to the dogma that OxA is a relatively stable end product of metabolism. Glycolate is a poor substrate for synthesis of OxA and CaOx formation, further establishing AsA as th immediate precursor in the synthesis of OxA used for calcium precipitation in crystal idioblasts.     

The Effects of Silicon on Cucumber Plants Grown in Recirculating Nutrient Solution

Cucumber plants (Cucumis sativus) cv. Corona were grown in recirculatingnutrient solution containing either 10 mg l^–1 SiO₂ (lowSi) which was the level present in the water supply or givenan additional 100 mg l^–1 SiO₂ (high Si). Silicate wasdepleted from the solution when cucumbers were grown, but accumulatedwhen tomatoes were grown. Major effects on cucumber leaves ofadded Si were: increased rigidity of the mature leaves whichhad a rougher texture and were held more horizontally; theywere darker green and senescence was delayed. The mature highSi leaves acquired characteristics of leaves grown in a higherlight intensity, i.e. they had shorter petioles and an increasedfresh weight per unit area, dry weight per unit area, chlorophyllcontent, RuBPcarboxylase activity and soluble protein (all expressedper unit area of interveinal laminar tissue). Addition of Sidid not affect the final leaf area of the mature leaves butroot fresh weight and dry weight were increased. A pronouncedeffect of Si addition was the increased resistance to the powderymildew fungus Sphaerotheca fuliginea. Despite regular applicationsof fungicide, outbreaks of the fungal disease occurred on mostof the mature leaves on the low Si plants, while the high Siplants remained almost completely free of symptoms. The additionof Si could be beneficial to cucumbers grown in areas wherethe local water supply is low in this element, especially whengrown in recirculating solution or in a medium low in Si, e.g.peat. Cucumber, silicon, fungal resistance, powdery mildew, senescence, RuBPcarboxylase     

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.