The Ultrastructure and Analytical Microscopy of Silicon Deposition in the Intercellular Spaces of the Roots of Molinia caerulea (L.) Moench

Silicon accumulation in mature roots of Molinia caerulea (L.)Moench. was investigated by transmission electron and Cora analyticalmicroscopy. Deposits are located in the intercellular spaces(ICS) of the persistent cortical cells immediately externalto the endodermis. The earliest deposits are in the form ofspheres lining the cavities and are followed by further depositswhich eventually fill the ICS. Evidence is presented that priorto and during the earliest silicon formations, many ICS revealedthe presence of cell organelles. No silicon was detected inthe inner tangential walls of the endodermis. The results are discussed in relation to previous studies ofsilicon deposits in the roots of Molinia and to possible pathwaysresulting in deposits associated with the ICS. Molinia caerulea(L.) Moench, roots, silicon deposition, analytical microscopy     

The Ultrastructure and Analytical Microscopy of Silicon Deposits in the Roots of Saccharum officinarum (L.)

Silicon accumulation in the endodermis of the ‘set’and ‘shoot’ roots of Saccharum officinarum (L.)were investigated by scanning electron microscopy and electron-probemicroanalysis. Silicon microassay was also carried out by meansof the Corinth analytical microscope (CORA). Aggregates arelargely associated with the inner tangential wall (ITW) of theendodermis and their formation is basically similar to thoseseen in Sorghum bicolor (L.) Moench. In contrast to Sorghumthe earliest deposits in Saccharum appear in wall strata wellwithin the cell wall cytoplasm interface. An additional layerof silicon was also located along the endodermal pericycle boundaryextending some distance along the middle lamella of the radialwalls. The results are discussed in relation to those of previous studiesof silicon accumulation in endodermal cells and to possiblefactors affecting such accumulations.     

X-ray Microanalysis of the Seminal Root of Sorghum bicolor with Particular Reference to Silicon

Sorghum bicolor (L.) Moench. cv. P508.GB plants were grown inwater culture for 1 week, when the seminal roots were harvestedand sampled at five positions starting from the base: 0.0, 0.25,0.50 and 0.75 of the axis length, and a sub-apical position,11 mm behind the tip. Mineral distribution in bulk frozen rootsegments was investigated using SEM and X-ray microanalysis.The elements detected were potassium, chlorine, sulphur, sodium,phosphorus, calcium and silicon. The first four occurred inall root zones. Phosphorus was ubiquitous, but appeared to accumulatein the pericycle protoplasm. Calcium and silicon exhibited themost variation along the seminal axis. Calcium was present inall tissues at the base, but decreased acropetally, being detectedin only the outer cortical and epidermal walls of the subapicalzone. Silicon was present at low levels in protoplasts and wallsof most root tissues, and accumulated in the endodermal protoplastand walls. Deposition in walls is initiated coincident withthe earliest stages of secondary wall thickening. Silicon contentof the inner tangential wall of the endodermis exhibits a decreasingacropetal gradient along the axis length. It is absent frommost cell walls of the sub-apical zone. Silicon pathways inthe root, and silica aggregate formation in relation to thesurrounding ionic environment, are discussed. Sorghum bicolor (L). Moench, seminal root, cryostage, SEM, X-ray microanalysis, ion localization, silicon, endodermis     

Endodermal Silicon Deposits and Their Linear Distribution in Developing Roots of Sorghum bicolor (L.) Moench.

Seedlings of Sorghum bicolor (L.) Moench. were cultured in nutrientsolution containing 100 p.p.m. SiO₂. Over periods up to 14 days,the progressive accumulation of Si deposits was recorded bymeans of the electron-probe microanalyser and the scanning electronmicroscope along the seminal root length. An acropetal, linear gradient of Si deposits developed in theendodermis, beginning at the proximal end of the root after1 day, and subsequently extended distally until after 7 days,the total silicified endodermal zone occupied the proximal 75per cent of the root length, a value which remained relativelyconstant thereafter, in spite of root extension. No Si was detectedbelow this zone towards the apex. This result is believed tobe related directly to the asynchronous gradient of cell maturationexhibited by the endodermis behind the apex, and specificallyto the degree of wall development therein. Opaline silica was deposited only in the endodermis, initiallyon the inner tangential wall (ITW) surface after only 1 day,as spherical masses of coalesced, primary particles for whichthe term ‘silica aggregate’ is proposed. A thinlayer of silica over the wall surface was formed as a secondaryphase. The aggregates reached mature size after approximately7 days. Conditions favourable to the inception of silica depositionare discussed including the significance of the chemical compositionof the aggregates, and the importance of the degree of cellulosicthickening, as well as the surface characteristics, of the ITW.     

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     

Endodermal Silicification in Mature, Nodal Roots of Sorghum bicolor (L.) Moench.

Nodal roots of mature, soil-grown specimens of Sorghum bicolor(L.) Moench. were investigated for their Si content by meansof the electron-probe microanalyser and the scanning electronmicroscope. No consistent accumulation of Si occurred other than in theendodermis. Indication of a lower Si content in the old as comparedto the younger nodal roots was obtained. The solid silica depositsoccurred as domeshaped silica aggregates confined to the innertangential wall. Comparisons with the similar deposits of youngseminal roots are made. It is argued that the endodermal deposits represent a specializedaspect of silicification and contrasting hypotheses to accountfor it are proposed, one involving physico-chemical factors,the other, protoplasmic control.     

Nitrate Reductase Activity: A Biochemical Criterion of Hybrid Vigour in Sorghum bicolor (L.) Moench

F₁ hybrids of Sorghum bicolor (L.) Moench and their inbred parentswere analysed for NADH-nitrate reductase activity during theearly stages of seedling growth. In all the hybrids both mid-parentaland better parental heterosis were discernible in shoots whereasin roots two hybrids out of the three tested, showed heteroticlevels. It is suggested that in sorghum nitrate reductase activityduring seedling stages can be used as a biochemical criterionfor evaluating hybrid vigour. Sorghum bicolor (L.) Moench, sorghum, hybrid vigour, nitrate reductase     

The Ultrastructure and Electron-probe Microassay of Silicon Deposits in the Endodermis of the Seminal Roots of Sorghum bicolor (L.) Moench

Si deposits in the endodermis of the seminal root of Sorghumbicolor (L.) Moench., following culture in nutrient solutioncontaining 100 ppm SiO₂ for 7 days, were investigated by transmissionelectron microscopy. Si microassay was carried out by meansof the EMMA-4 system. Endodermal ultrastructure is discussed. EM micrographs of theSi deposits reveal information regarding their formative processesand indicate a considerable involvement with the cellulosicstructure of the inner tangential wall (ITW). The initial depositsare believed to be composed structurally of particles designatedas primary spherical units. The EMMA-4 conclusively indicated that Si was localized andconfined to the ITW, and the implications of this result forprevious studies are considered. Current evidence from studies of endodermal function, as wellas root translocation physiology, is utilized in arriving attentative mechanisms for silicic acid transport and subsequentSi deposition on the ITW.     

The Interaction Between Silicon and Aluminium in Sorghum bicolor (L.) Moench: Growth Analysis and X-ray Microanalysis

Seeds of Sorghum bicolor (L.) Moench. were germinated on moistfilter paper for 6 d, before the seedlings were transferredto pots containing 500 µmol l^-1 Ca(NO₃)₂ for 2 d. Theseedlings were then treated with 0 or 100 µmol l^-1 Alin factorial combination with 0, 1400 or 2800 µmol l^-1Si for 8 d. The background solution used throughout was 500µmol l^-1 Ca(NO₃)₂. Aluminium treatment reduced root growthand caused a significant increase in shoot/root ratio. The presenceof silica in the solution significantly ameliorated the effectsof aluminium on root growth. Three treatment were selected for a microanalytical investigationof the basal region of the root: 2800 µmol l^-1 Si only;100 µmol l^-1 Al only; and a combination of the two. Inthe 2800 µmol l^-1 treatment silica was deposited in theendodermis, with the greatest accumulation being in the innertangential wall (ITW). When plants were treated with 100 µmoll^-1 Al only, aluminium concentration was highest in the outertangential wall (OTW) of the epidermis. The element was presentin the hypodermal walls and OTW of the endodermis and was notdetectable in the stele. With both 2800 µmol l^-1 Si and100 µmol l^-1 Al in the nutrient solution the two biomineralizationsites were the ITW of the endodermis, where silicon was themajor element deposited, and atypically in the OTW of the epidermiswhere both aluminium and silicon were present. The sequestrationof aluminium in the Al-Si deposit in the OTW of the epidermismay represent the mechanism that allows greater root growthin this treatment.Copyright 1993, 1999 Academic Press Sorghum bicolor (L.) Moench., aluminium, silicon, calcium, root, toxicity, biomineralization, X-ray microanalysis, freeze substitution     

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     

Leaf Abscission Zones in Molinia caerulea (L.) Moench, the Purple Moor Grass

Leaf separation and loss in the grass Molinia caerulea (L.)Moench was investigated using scanning electron microscopy.Leaf senescence and subsequent shedding of leaves was precededby the formation of a leaf abscission zone. The zone containscells which have more than doubled their wall thickness to greaterthan 0-4 µm. The line of fracture associated with thezone principally followed the middle lamellae, leaving intactcells on the fracture faces. Molinia should prove an interestingmodel in which to study abscission processes in the Gramineae. Molinia caerulea, leaf abscission zone, electron microscopy     

Variation in Molinia caerulea (L.) Moench, the Purple Moor Grass, in Relation to Edaphic Environments

Two diverse population of Molinia caerulea (L.) Moench, onegrowing on a Ca²⁺-rich, alkaline, Leblanc waste tip, the otheron an acid moorland, were investigated. It was anticipated thata comparative study of the morphology, growth and floweringpatterns, protein composition, specific isoenzyme and root-surfacephosphatase activities of both populations would clarify thetaxonomic status of populations of Molinia and help towardsunderstanding the ability of M. caerulea to colonize widelydiffering habitats. Both populations were identified as Moliniacaerulea caerulea. The Molinia growing on Leblanc spoil displayedcharacteristics typified by r-selected plants, i.e. displayinglarger flowers after faster development. Acid moorland plantsshowed greater vegetative development and were more typicalof K-selected types. These differences between the more vigorousshoot growth of the moorland compared to the waste tip plantspersisted under controlled conditions, irrespective of the mediain which they were grown. Root surface phosphatase activityshowed a plastic response to edaphic pH which may contributeto the success of M. caerulea caerulea in colonizing diversehabitats.Copyright 1995, 1999 Academic Press Molinia, edaphic extremes, variation, morphometry, enzyme activities, plasticity, colonization, taxonomy     

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.     

Inhibition of Anthocyanin Synthesis by Cobaltous Ions in the First Internode of Sorghum bicolor L. Moench

Cobaltous ions (Co²⁺) inhibited light-mediated anthocyanin synthesisin excised first internodes of Sorghum bicolor (L.) Moench.Studies with precursors/intermediates of the anthocyanin biosyntheticpathway, such as acetate, phenylalanine, t-cinnamic acid, tyrosine,and p-coumaric acid were undertaken to identify the metabolicsite at which Co²⁺ ions inhibit anthocyanin synthesis. p-Coumaricacid partially restored anthocyanin synthesis. No other intermediatewas effective in bringing about recovery. It is suggested thatCo²⁺ might interfere with process/es which leads to the formationof p-coumaric acid, an intermediate of the anthocyanin biosyntheticpathway. Key words: Sorghum bicolor (L.) Moench, anthocyanin, cobaltous ions, phenylalanine ammonia-lyase     

Viability and Germination of the Pollen of Sorghum [Sorghum bicolor (L.) Moench]

Previous studies have demonstrated that pollen of sorghum [Sorghumbicolor (L.) Moench] loses capacity to both germinate in vitroand to set seed in vivo soon after being shed. The current studyevaluates the capacity for dehydrated pollen to effect in vitrogermination, reduce tetrazolium chloride, and set seed on cytoplasmicmale sterile plants. Morphological changes during pollen germinationwere examined by scanning electron microscopy (SEM). Close to70% of the pollen germinated in 5 min, or less, when collectedat 80% relative humidity (RH) and stored in sealed glass vials.Pollen tubes elongated autotropically with atmospheric humidityapparently being a controlling factor in the process. Pollendehydrated at 50% RH and 25°C for 15-30 min neither germinatedin vitro, reduced tetrazolium chloride, nor set seed on malesterile plants. Rehydrating the pollen did not restore the capacityfor germination. SEM micrographs demonstrated that elongatingpollen tubes encircled the pollen grain and were contiguousto the surface. A fibrillar-like material existed on the exineof separated pollen grains at the point where the grains hadbeen previously attached.Copyright 1994, 1999 Academic Press Sorghum pollen, germination, seed-set, viability, scanning electron microscopy, Sorghum bicolor (L.) Moench     

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.     

OBSERVATIONS ON THE ROOT ANATOMY OF RICE (ORYZA SATIVA L.)

Rice plants were grown hydroponically and roots were prepared for light and electron microscopy using standard techniques. The roots are bounded by an epidermis, exodermis, and fibrous layer. The exodermis has a suberin lamella along its inner tangential wall. The fibrous layer is composed of thick-walled lignified cells with little pitting. The cortical parenchyma is compact when young, but expands and separates to form a zone of cell walls and air spaces in a spoked arrangement. Supporting columns of living parenchyma cells are occasionally present, particularly near lateral roots. The endodermis is typical for grasses with Casparian strips, suberin lamellae, and tertiary state walls with numerous pits. The pericycle and pith become sclerified. Protoxylem elements alternate with protophloem in the young root; later, early metaxylem, late metaxylem, and metaphloem proliferate. The exodermis, fibrous layer, lacunate cortex, and endodermis appear to present a formidable barrier to radial ion movement in the mature portions of the root.     

The Distribution of Silicon Deposits in the Fronds of Pteridium aquilinum L.

The occurrence of silicon in mature fronds of Pteridium aquilinumwas investigated using light and scanning electron microscopyand X-ray micro analysis. The heaviest deposits were locatedin the outer tangential walls of the epidermis of the petiole,but the upper epidermal cells of the costal regions of the laminaealso accumulated small quantities. No discrete bodies were associatedwith these deposits. However, the mid-ribs of the laminae exhibitedsome long siliceous fibres associated with the luminae of thestrengthening tissues. The significance of these accumulationsin relation to deposition mechanisms and possible carcinogeniceffects are discussed. Pteridium aquilinum L., bracken, frond, microanalysis, silica     

SILICON IN THE ROOTS OF HIGHER PLANTS

Silicon (Si) distribution in the roots of Sorghastrum nutans (L.) Nash and Sorghum bicolor (L.) Moench. was investigated by means of the electron-probe microanalyzer and scanning electron microscope. In both species, Si was confined to the inner tangential wall of the tertiary-phase endodermal cells in the form of nodular silica aggregates of similar morphology and X-ray intensity. The results are compared to those for six closely related genera, as well as to studies of Si in the roots of species of other tribes of the family Poaceae. The various types of root deposits occurring in the family are described, and their relationships discussed. It is concluded that the type of Si distribution exhibited is determined largely by the phylogenetic status of the genus, rather than by the basic pattern of root anatomy.     

Electron-probe Microanalysis of Silicon in the Roots of Oryza sativa L.

Employing the electron-probe microanalyser, the localizationof silicon in the different tissues of the mature root of ricehas been investigated. Deposition of silicon is specific tothe endodermis. The heaviest accumulation is associated withthe inner tangential walls and to some extent the radial wallsof the endodermal cells.