Scanning Electron Microscopical Observation and X-ray Microanalysis of Erysiphe pisi DC on Infected Leaves of Pea (Pisum sativum L)

Observations on scanning electron microscope (SEM) and low vacuum scanning electron microscope (LVSEM) of Erysiphe pisi in infected leaves of pea (Pisum sativum). are reported together with data from X-ray micro-analysis. Morphology of ungerminated and germinated conidia was observed and LVSEM revealed halo formation around the penetration peg of the pathogen. X-ray microanalysis of the composition of halo showed the accumulation of Si and P. Presence of S. Mg, Cl, K and Ca was also discerned. Deposition of Si in the halo and profuse growth of the pathogen indicate that Si deposition is not an essential signal for inducing resistance in the host. LVSEM appears to be a suitable and rapid method to study the morphology as well as halo formation in E. pisi. X-ray microanalysis and LVSEM studies of E. pisi are reported for the first time.     

应用电子探针对植物根际和根内营养元素微区分布的探讨

用电子探针可检测出玉米、大豆根际和根内含有Na,Mg,Al,Si,P,S,Cl,K,Ca,Ti,Fe,Cu和Zn 13种元素。这些元素在根际土壤、粘液层和根组织内的含量分布有一定的规律性。除Si,Al,Ca,Fe在根际土壤中峰值较高外,Ti仅在土壤中达到可检测量;S,Fe和Zn富集在粘液层,Mg,P,Cl只在根组织内才有较明显的峰。这些规律可作为区分根—土界面的参考指标。K含量在根内明显高于根际土壤,并由表皮层到中柱径向增加;Ca则与K不同,且受植物种类的影响。     

Mobility and deposition of silicon in cucumber plants

Abstract. The mobility and deposition of silicon (Si) in cucumber plants grown in hydroponic culture were studied using scanning electron microscopy and energy dispersive X-ray analysis under conditions of powdery mildew fungus infection. Following transfer to Si containing media, plants grown in the absence of Si showed rapid silicification of leafy tissue, primarily in the trichome bases. These plants also exhibited increased disease resistance to Sphaerotheca fuliginea (Schlecht.:Fr.) Poll, and concentration of Si in the leaf epidermis surrounding the invading pathogen. Plants grown in Si-supplemented media and transferred into Si-deficient media contained residual Si in the leaf trichome bases, but failed to display disease resistance or silicification of host tissue surrounding the invading pathogen.     

High energy X-ray scattering tomography applied to bone

High energy synchrotron X-ray scattering was developed for reconstruction of specimen cross-sections. The technique was applied to a model specimen of cortical bone containing a capillary tube of silicon, and reconstructions were produced with either full diffraction rings or texture-related subsets of a given ring. The carbonated apatite (cAp) 00.2 and 22.2 reconstructions and the Si 311 reconstructions agreed with absorption-based reconstructions from the measured X-ray transmissivity recorded during diffraction pattern acquisition and from reconstructions produced subsequently of the same specimen using a commercial microCT (microComputed Tomography) scanner.     

Elemental composition of the cyanobacterium Anabaena flos-aquae collected from different depths within a stratified lake

The elemental composition of cells of Anabaena flos-aquae was investigated by energy-dispersive electron probe X-ray microanalysis (10?kV) of mixed phytoplankton samples obtained from different depths within a stratified eutrophic freshwater lake. Preliminary studies had indicated that at 10?kV, X-ray information was derived mainly from the peripheral region (up to 1–2?μm) of cells. Routinely detectable elements (present in at least 50% of cells) included Mg (overall mean 112?mmol?kg^?1 dry mass), Si (1850), P(313), S (113), Cl (92), K (300) and Ca (56). Peaks of Na and Al were also frequently present. Individual elements showed a wide range of intracellular concentrations at each depth within the lake. Si had a clear bimodal distribution at depths of 1–8?m, indicating both low-Si (mean concentrations 54–290–3800?mmol?Si?kg^?1) and high-Si (2770–3800?mmol?Si?kg^?1) cell subpopulations. Low-Si cells had significantly higher concentrations of other detectable elements compared with high-Si cells, but could not be distinguished from the latter in terms of morphology or stage of cell cycle (comparison of dividing and non-dividing cells). High-Si cells were intermixed with low-Si cells in individual Anabaena colonies and decreased proportionally with depth, ranging from approximately 75% in the surface sample to 10% at 8?m. With the exception of Si, mean elemental concentrations of Anabaena populations were closely similar throughout the epilimnion. These differed from the metalimnion sample, which had significantly higher concentrations of Mg, P, S and K, and a lower concentration of Si. Mean ratios of certain elements (Mg, P, S and K) and ion groups (monovalent/divalent cations) were highly constant throughout the sampled water column. At each depth, most of the cell elements were significantly and positively inter-correlated (Pearson analysis), with Mg, P, S and K typically constituting the major group (factor analysis). Correlations with Si were invariably negative, suggesting a separate (possibly cell wall) location of this element in the cell.     

The Effects of Silicon Supplementation on Cucumber Fruit: Changes in Surface Characteristics

Addition of silicate to hydroponic nutrient media has been shownto increase resistance of cucumber to powdery mildew. As a sideeffect of this treatment, the fruit surface developed an unusualdull appearance, or bloom. The effects of silicate supplementationon cucumber fruit were observed using scanning and transmissionelectron microscopy, energy-dispersive X-ray analysis, and colourimetricassays. Changes in the fruit trichome morphology occurred: trichomesfrom + Si fruit had a coarse outer appearance compared to -Si fruit where the trichomes were smooth. X-ray mapping showedhigh Si content in trichomes but not in the surrounding epidermisor fleshy mesocarp and endocarp tissues. Sections through theepidermal layer reveal that the silica is restricted to thetrichomes, primarily in the epicuticular wax.Copyright 1993,1999 Academic Press Cucumber, Cucumis sativus L., fruit, silica, energy dispersive X-ray analysis, trichomes     

Clay minerals as a soil potassium reservoir: observation and quantification through X-ray diffraction

Potassium (K) is a major element for plant growth. The K⁺ ions fixed in soil 2:1 clay mineral interlayers contribute to plant K nutrition. Such clay minerals are most often the majority in temperate soils. Field and laboratory observations based on X-ray diffraction techniques suggest that 2:1 clay minerals behave as a K reservoir. The present work investigated this idea through data from a replicated long term fertilization experiment which allowed one to address the following questions: (1) Do fertilization treatments induce some modifications (as seen from X-ray diffraction measurements) on soil 2:1 clay mineralogy? (2) Are soil 2:1 clay mineral modifications related to soil K budget in the different plots? (3) Do fertilizer treatments modify clay Al, Si, Mg, Fe or K elemental content? (4) Are clay mineral modifications related to clay K content modifications? (5) Are clay mineral changes related to clay Al, Si, Mg or Fe content as well as those of K content? Our results showed that K fertilization treatments considered in the context of soil K budget are very significantly related to 2:1 soil clay mineralogy and clay K content. The 2:1 clay mineral modifications observed through X-ray measurements were quantitatively correlated with chemically analyzed clay K content. Clay K content modifications are independent from clay Al, Si, Mg or Fe contents. These results show that the soil chemical environment can modify interlayer site occupations (illite content) which suggests that high level accumulation of potassium can occur without any modification of the clay sheet structure. This study therefore validates the view of 2:1 clay minerals as a K reservoir easily quantifiable through X-ray observations.     

Silicon-containing granules of rat liver,kidney and spleen mitochondria

Summary Isolated rat liver mitochondria containing granule aggregates (25–75 nm in diameter) and small (5–10 nm) electron opaque granules were examined by electron probe X-ray microanalysis. The granule aggregates gave an intense Si signal, while the small granules gave both Si and P signals. Isolated mitochondria of rat liver, spleen and kidney, subjected to detergent solubilization and differential centrifugation, produced two granule fractions: (1) a 10,000g fraction consisting predominantly of granule aggregates (25–75 nm) composed of smaller granules (5–10 nm in diameter), and (2) a 10,000–30,000 g fraction of non-aggregated small granules (5–10 nm). Thin sections of isolated granule aggregates gave Si X-ray signals similar to those obtained from in situ granules. In addition S, Cl, Mg, Cr and Fe X-ray signals were observed. Cr occurred only in the large kidney granules, while Fe occurred in both fractions of the spleen and kidney granules. The presence of Si in the granules was confirmed by chemical analysis of the isolated granules and in vivo radiolabeling of the granules with ³¹Si and ⁶⁸Ge. Contamination within the electron microscope was eliminated by a liquid nitrogen anticontamination device.Supported by Grant GM-08229-13-15 from the National Institutes of Health, USPHSWe are grateful to the Perkin-Elmer Corporation and to their Western Branch Manager, Mr. Michael E. Mullen and Microscopist, Mr. Minoru Saito, for use of and assistance with the Hitachi H 500 transmission electron microscope with the scanning attachment, and to the Kevex Corporation for the use of the Kevex X-ray Spectrometer. We also wish to acknowledge Mary Louise Chiappino for her technical assistance in preparing the thin sections, the final micrographs and X-ray spectra photographs and Darlene Lum for technical assistance in the laboratory     

Rapid and accurate analyses of silicon and phosphorus in plants using a portable X-ray fluorescence spectrometer

The elemental analysis of plant material is a frequently employed tool across biological disciplines, yet accurate, convenient and economical methods for the determination of some important elements are currently lacking. For instance, digestion-based techniques are often hazardous and time-consuming and, particularly in the case of silicon (Si), can suffer from low accuracy due to incomplete solubilization and potential volatilization, whilst other methods may require large, expensive and specialised equipment. Here, we present a rapid, safe and accurate procedure for the simultaneous, nonconsumptive analysis of Si and phosphorus (P) in as little as 0.1 g dried and ground plant material using a portable X-ray fluorescence spectrometer (P-XRF). We used certified reference materials from different plant species to test the analytical performance of P-XRF and show that the analysis suffers from very little bias and that the repeatability precision of the measurements is as good as or better than that of other methods. Using this technique we were able to process and analyse 200 ground samples a day, so P-XRF could provide a particularly valuable tool for plant biologists requiring the simultaneous nonconsumptive analysis of multiple elements, including those known to be difficult to measure such as Si, in large numbers of samples.     

Silicon in rat liver organelles: Electron probe microanalysis

Summary Electron probe microanalysis of unfixed freeze-substituted rat liver tissue embedded in Spurr's low viscosity epoxy resin demonstrated the occurrence of Si as well as P, S, and Cl in the nucleus, nucleolus, mitochondria, and rough endoplasmic reticulum. Chemical analysis confirmed that the Si in the organelles did not originate from instrumental contaminants. This suggests that Si may be involved in the biochemistry of these subcellular organelles.Supported by Grant GM-08229-12-13 from the National Institutes of Health, USPHS.We are grateful to the Kevex Corporation and Mr. Glenn W. Meyer, Sales Engineer, for the use of the Kevex X-ray spectrometer; we wish to thank as well the Perkin-Elmer Corporation and their Western Branch Manager, Mr. Michael E. Mullen and Senior Microscopist, Mr. Minoru Shinorhara, for use of and assistance with the Hitachi HU 12 A transmission electron microscope. We also wish to acknowledge Mary Louise Chiappino for her technical assistance in preparing the thin sections, the final micrographs and the X-ray photographs, and Darlene Lum for technical assistance in the laboratory.     

Silica deposition in Demosponges: spiculogenesis in Crambe crambe

Transmission electron-microscopy images coupled with dispersive X-ray analysis of the species Crambe crambe have provided information on the process of silica deposition in Demosponges. Sclerocytes (megasclerocytes) lie close to spicules or surround them at different stages of growth by means of long thin enveloping pseudopodia. Axial filaments occur free in the mesohyl, in close contact with sclerocytes, and are triangular in cross section, with an internal silicified core. The unit-type membrane surrounding the growing spicule coalesces with the plasmalemma. The axial filament of a growing spicule and that of a mature spicule contain 50%-70% Si and 30%-40% Si relative to that contained in the spicule wall, respectively. The extracellular space between the sclerocyte and the growing spicule contains 50%-65%. Mitochondria, vesicles and dense inclusions of sclerocytes exhibit less than 10%. The cytoplasm close to the growing spicule and that far from the growing spicule contain up to 50% and less than 10%, respectively. No Si has been detected in other parts of the sponge. The megascleres are formed extracellularly. Once the axial filament is extruded to the mesohyl, silicification is accomplished in an extracellular space formed by the enveloping pseudopodia of the sclerocyte. Si deposition starts at regularly distributed sites along the axial filament; this may be related to the highly hydroxylated zones of the silicatein-alpha protein. Si is concentrated in the cytoplasm of the sclerocyte close to the plasmalemma that surrounds the growing spicules. Orthosilicic acid seems to be pumped, both from the mesohyl to the sclerocyte and from the sclerocyte to the extracellular pocket containing the growing spicule, via the plasmalemma.     

DIATOM MINERALIZATION OF SILICIC ACID. II. REGULATION OF Si (OH)4 TRANSPORT RATES DURING THE CELL CYCLE OF NAVICULA PELLICULOSA1

Synchronized populations of Navicula pelliculosa (Bréb.) Hilse show a 10-fold increase in Si(OH)₄ transport rate during traverse through the cell division cycle. The transport activity pattern is similar to a “peak enzyme.” Kinetic analysis showed there was a significant change in K_s values, indicating increased “affinity” for Si(OH)₄ as cells neared maximal uptake rates. However, the dramatic changes in transport rate at various cell cycle stages were also reflected by alterations in the V_max, values of the transport process, suggesting a change in the number of functional transport “sites” in the plasma membrane. Cells in the wall forming stage, arrested from further development by Si(OH)₄ deprivation, maintained high transport rates for as long as 7 h. The rates decreased rapidly if protein synthesis were blocked or if Si(OH)₄ was added, the latter allowing the cells to traverse the rest of the cycle. The half-life of the transport activity ranged from 1.0 to 2.2 h when protein synthesis was inhibited at various cell cycle stages and during the natural decline of activity late in the cycle. The transport system appears to be metabolically unstable as is typical for a “peak protein.” The rise in transport rate through the cell cycle did not depend on the presence of Si(OH)₄ in the medium; therefore, the transport system does not appear to be induced by its substrate. The rise in transport is also observed in L:D synchronized cells developing in the presence of Si(OH)₄; neither does the transport system appear to be derepressed. The transport rate was strongly cell cycle-stage dependent; the data appeared to fit the “dependent pathway” model proposed by Hart-well to explain oscillations in enzyme synthesis during the cell cycle.     

In Operando Strain Measurement of Bicontinuous Silicon‐Coated Nickel Inverse Opal Anodes for Li‐Ion Batteries

Elastic strains are measured in operando in a nanostructured silicon‐coated nickel inverse opal scaffold anode, using X‐ray diffraction to study the Si (de)lithiation‐induced Ni strains. The volume expansion upon lithiation of the Si in the anode is constrained by the surrounding Ni scaffold, causing mismatch stresses and strains in the Si and Ni phases during cycling. The Ni strains are measured in operando during (dis)charge cycles, using diffraction peak position and peak broadness to describe the distribution of strain in the Ni. During lithiation, compressive strains in the Ni first increase linearly with charge, after which a gradually decreasing strain rate is observed as the maximum lithiation state is approached; upon delithiation a similar process occurs. In‐plane average compressive strains on the order of 990 ± 40 με are measured in the Ni scaffold during lithiation, corresponding to compressive stresses of 215 ± 9 MPa. The decreasing strain rates and decreasing maximum and recovered strains suggest that plasticity in Ni and/or Si, as well as delamination between Ni and Si, may occur during cycling. Rate sensitivity in capacity is correlated with strain and a maximum Ni compressive stress of 230 ± 40 MPa is measured at the maximum state of lithiation.     

Electron-probe microanalysis of silicon in the epidermis of rice (Oryza sativa L.) internodes

Summary Electron-probe X-ray microanalysis showed that significant amounts of silicon are accumulated in the entire epidermal system of the rice internode except in the stomatal apparatuses. Thus, there is a lack of specific sites for Si deposition from levels just above the base to the tip of the rice internode. In the intercalary meristem region, 1 cm above the base of the internode, point-count data indicate more Si accumulation in the dumb-bell shaped silica cells than in the long epidermal cells. Above this region, Si is accumulated essentially in a uniform pattern in all epidermal cells. Such a pattern for Si accumulation in rice internodes markedly contrasts with that for Avena internodes and may explain, in part, why rice plants have a higher percentage Si (dry weight basis) in their shoots. The adaptive significance of this silicification pattern in rice is discussed.     

Elemental composition in vesicles of an arbuscular mycorrhizal fungus, as revealed by PIXE analysis

We investigated element accumulation in vesicles of the arbuscular mycorrhizal (AM) fungus Glomus intraradices, extracted from the roots of inoculated leek plants. The elemental composition (elements heavier than Mg) was quantified using particle-induced X-ray emission (PIXE), in combination with scanning transmission ion microscopy (STIM). In vesicles, P was the most abundant of the elements analysed, followed by Ca, S, Si and K. We analysed 12 vesicles from two root systems and found that the variation between vesicles was particularly high for P and Si. The P content related positively to Si, Zn and K, while its relation to Cl fitted to a negative power function. Vesicle transects showed that P and K were present in central parts, while Ca was present mainly near the vesicle surfaces. The results showed that P is an important part (0.5% of the dry weight) of the vesicle content and that the distribution of some elements, within mycelia, may be strongly correlated.     

Changes in the elemental composition of Asterionella formosa during the diatom spring bloom

Changes in the elemental composition of the diatom Asterionellaformosa within mixed phytoplankton samples were determined overthe spring bloom period using energy dispersive X-ray microanalysis.Using a 10 kV electron microprobe, X-ray information from thetop 1–2 µm of the cell revealed overall mean concentrationsof: Si (4636 mmol kg^–1 dry wt), P (82), S (54), Cl (71)K (94) and Ca (90). Concentrations of all elements showed widevariation within each date sample, with unimodal frequency distributionsapproximating to a Normal distribution. Correlation of the entiredata set demonstrated clear statistical associations betweenelements, including Si, P and K. Uptake of Si during the courseof the diatom bloom led to a major fall in lake water concentration(1.0 to 0.07 mg l^–1), which correlated with a decreasein the mean cell Si concentration (6735 to 3107 mmol kg^–1dry wt) during the final phase of the bloom. Mean cell concentrationsof P and K also showed marked decreases with time, in closeparallel with cell Si. These changes in P and K were attributedto the high level of internal correlation with Si and not toany significant decrease in P and K availability in the environment.     

施硅增强水稻对纹枯病的抗性

采用水培的方法,从细胞学和生理生化方面研究了硅增强水稻对纹枯病的抗性作用。结果表明：加硅处理的水稻叶片硅化细胞和叶片表面的硅元素含量均显著高于缺硅处理（对照）：接种纹枯病菌后,加硅处理的MDA含量总体上低于缺硅处理,峰值尤为显著;加硅处理的SOD活性始终高于缺硅处理,接种后第4天加硅处理SOD活性较低时,其POD活性较高,而缺硅处理的POD活性较低,表明硅增强了SOD和POD之间的协调性;接种后硅对CAT和PAL活性没有产生明显影响,但降低了PP0活性;加硅能显著降低水稻植株的纹枯病病情指数。     

根足类原生动物半圆表壳虫壳体生物矿化特征

以有壳阿米巴类原生动物 (肉鞭门 ,叶足纲 )半圆表壳虫 (Arcellahemisphaerica)为研究对象 ,利用光镜和扫描电镜研究了其壳体生物矿化的特征。结果显示 :矿化前期 ,壳体为无色透明且柔软易变形 ;中期 ,为黄色较坚硬 ;后期 ,为褐色坚硬。通过X 射线显微分析术鉴定不同矿化时期壳体的无机元素 ,结果表明 ,与矿化前期相比矿化中期和后期壳体中Si、Mn和Fe的比例增加 ,Cl、K、S和Na的比例减少。由此推测 ,⑴半圆表壳虫矿化过程中壳体坚硬的原因是Si、Mn和Fe的比例增加导致 ,黄色是由微量Mn和Fe引起 ,褐色是高含量Mn与低含量Fe的反映 ;⑵半圆表壳虫壳体矿化以构成壳体的小泡为基本单位进行 ,Si、Mn和Fe矿化体以氧化物形式通过分子间相互作用力 ,与小泡壁上氨基酸或多肽的羟基作用 ,自组装合成。     

Organisation of subunits in chromatin.

There is considerable current interest in the organisation of nucleosomes in chromatin. A strong X-ray and neutron semi-meridional diffraction peak at approximately 10 nm had previously been attributed to the interparticle specing of a linear array of nucleosomes. This diffraction peak could also result from a close packed helical array of nucleosomes. A direct test of these proposals is whether the 10 nm peak is truly meridional as would be expected for a linear array of nucleosomes or is slightly off the meridian as expected for a helical array. Neutron diffraction studies of H1-depleted chromatin support the latter alternative. The 10 nm peak has maxima which form a cross-pattern with semi-meridional angle of 8 to 9 degrees. This is consistent with a coil of nucleosomes of pitch 10 nm and outer diameter of approximately 30 nm. These dimensions correspond to about six nucleosomes per turn of the coli.     

Adsorption of Adenine and Thymine on Zeolites: FT-IR and EPR Spectroscopy and X-Ray Diffractometry and SEM Studies

The interactions of adenine and thymine with and adsorption on zeolites were studied using different techniques. There were two main findings. First, as shown by X-ray diffractometry, thymine increased the decomposition of the zeolites (Y, ZSM-5) while adenine prevented it. Second, zeolite Y adsorbed almost the same amount of adenine and thymine, thus both nucleic acid bases could be protected from hydrolysis and UV radiation and could be available for molecular evolution. The X-ray diffractometry and SEM showed that artificial seawater almost dissolved zeolite A. The adsorption of adenine on ZSM-5 zeolite was higher than that of thymine (Student-Newman-Keuls test-SNK p<0.05). Adenine was also more greatly adsorbed on ZSM-5 zeolite, when compared to other zeolites (SNK p<0.05). However the adsorption of thymine on different zeolites was not statistically different (SNK p>0.05). The adsorption of adenine and thymine on zeolites did not depend on pore size or Si/Al ratio and it was not explained only by electrostatic forces; rather van der Waals interactions should also be considered.