•Encrusted leaf structures affect leaf optical properties.•Elemental localization analysis shows that these structures are encrusted by Si and/or by Ca.•The Si and Ca contents and their distribution ...patterns differ among the species.•Reflectance spectra were significantly affected by the concentrations of Ca.•Si, Ca and Mg concentrations significantly affected transmittance spectra.
This study aimed to relate the properties of incrusted plant tissues and structures as well as biomineral concentrations and localization with leaf reflectance and transmittance spectra from 280nm to 880nm in the grasses Phragmites australis, Phalaris arundinacea, Molinia caerulea and Deschampsia cespitosa, and the sedge Carex elata. Redundancy analysis revealed that prickle-hair length on adaxial surface and thickness of lower epidermis exerted significant effects in P. australis; prickle-hair density at abaxial leaf surface and thickness of epidermis on adaxial leaf surface in P. arundinacea; thickness of epidermis on adaxial leaf in D. cespitosa; prickle-hair density on adaxial leaf surface and thickness of cuticle in M. caerulea; and prickle-hair density on adaxial leaf surface and cuticle thickness of the lower side in C. elata. Micro-PIXE and LEXRF elemental localization analysis show that all of these structures and tissues are encrusted by Si and/or by Ca. Reflectance spectra were significantly affected by the Ca concentrations, while Si and Mg concentrations and the Ca concentrations significantly affected transmittance spectra. High concentrations of Mg were detected in epidermal vacuoles of P. arundinacea, M. caerulea and D. cespitosa. Al co-localises with Si in the cuticle, epidermis and/or prickle hairs.
Crop Fe deficiency is a worldwide problem. The aim of this study was to assess the effects of foliar Fe applications in two species grown in different environments: peach (Prunus persica L. Batsch) ...trees grown in the field and sugar beet (Beta vulgaris L. cv. "Orbis") grown in hydroponics. The distal half of Fe-deficient, chlorotic leaves was treated with Fe sulfate by dipping and using a brush in peach trees and sugar beet plants, respectively. The re-greening of the distal (Fe-treated) and basal (untreated) leaf areas was monitored, and the nutrient and photosynthetic pigment composition of the two areas were also determined. Leaves were also studied using chlorophyll fluorescence imaging, low temperature-scanning electron microscopy microanalysis, scanning transmission ion microscopy-particle induced X-ray emission and Perls Fe staining. The distal, Fe-treated leaf parts of both species showed a significant increase in Fe concentrations (across the whole leaf volume) and marked re-greening, with significant increases in the concentrations of all photosynthetic pigments, as well as decreases in de-epoxidation of xanthophyll cycle carotenoids and increases in photochemical efficiency. In the basal, untreated leaf parts, Fe concentrations increased slightly, but little re-greening occurred. No changes in the concentrations of other nutrients were found. Foliar Fe fertilization was effective in re-greening treated leaf areas both in peach trees and sugar beet plants. Results indicate that the effects of foliar Fe-sulfate fertilization in Fe-deficient, chlorotic leaves were minor outside the leaf surface treated, indicating that Fe mobility within the leaf is a major constraint for full fertilizer effectiveness in crops where Fe-deficiency is established and leaf chlorosis occurs.
Micro-proton induced X-ray emission (micro-PIXE) method was applied to study the micro-localization of silver (Ag) in digestive glands of a terrestrial arthropod (Porcellio scaber) after feeding on ...silver nanoparticles (nano-Ag) dosed food. The aim of our work was to assess whether feeding on nano-Ag results in the assimilation of silver (Ag) in digestive gland cells. To study micro-localization and elemental distribution of Ag, the animals were fed on food dosed with nanoparticles for 14days under controlled laboratory conditions. At the end of the feeding exposure, the animals were dissected and digestive glands prepared for micro-PIXE analyses and TEM investigation. The results obtained by micro-PIXE documented high amounts of Ag inside S-cells of the digestive gland epithelium; however, TEM investigation did not show particle aggregates inside digestive gland cells. Also no adverse effect on feeding behavior was recorded what is a measure of toxic effects. We explain the presence of Ag inside the cells as a result of the assimilation of dissoluted Ag ions from ingested nano-Ag particles. Assimilation of excessive amounts of ingested metal ions in S-cells is a well known metal detoxification mechanism in isopods. We discuss the advantages of using micro-PIXE for the micro-localization of elements in biological tissue in studies of interactions between nanoparticles and biological systems.
Bulk element concentrations of whole grain and element spatial distributions at the tissue level were investigated in wheat (Triticum aestivum) grain grown in Zn-enriched soil. Inductively coupled ...plasma mass spectrometry and inductively coupled plasma optical emission spectrometry were used for bulk analysis, whereas micro-proton-induced X-ray emission was used to resolve the two-dimensional localization of the elements. Soil Zn application did not significantly affect the grain yield, but did significantly increase the grain Ca, Fe and Zn concentrations, and decrease the grain Na, P and Mo concentrations; bulk Mg, S, K, Mn, Cu, Cd and Pb concentrations remained unchanged. These changes observed in bulk element concentrations are the reflection of tissue-specific variations within the grain, revealing that Zn application to soil can lead to considerable alterations in the element distributions within the grain, which might ultimately influence the quality of the milling fractions. Spatially resolved investigations into the partitioning of the element concentrations identified the tissues with the highest element concentrations, which is of utmost importance for accurate prediction of element losses during the grain milling and polishing processes.
A better understanding of the mechanisms that govern copper (Cu) uptake, distribution and tolerance in Brassica carinata plants in the presence of chelators is needed before significant progress in ...chelate-assisted Cu phytoextraction can be made. The aims of this study were therefore to characterise (S,S)-N,N′-ethylenediamine disuccinic acid (EDDS)-assisted Cu uptake, and to compare the spatial distribution patterns of Cu in the roots and leaves of B. carinata plants. The plants were treated with 30μM or 150μM CuSO4 or CuEDDS in hydroponic solution. Quantitative Cu distribution maps and concentration profiles across root and leaf cross-sections of the desorbed plants were obtained by micro-proton induced X-ray emission. In roots, the 30μM treatments with both CuSO4 and CuEDDS resulted in higher Cu concentrations in epidermal/cortical regions. At 150μM CuSO4, Cu was mainly accumulated in root vascular bundles, whereas with 150μM CuEDDS, Cu was detected in endodermis and the adjacent inner cortical cell layer. Under all treatments, except with a H+-ATP-ase inhibitor, the Cu in leaves was localised mainly in vascular tissues. The incubation of plants with 150μM CuEDDS enhanced metal translocation to shoots, in comparison to the corresponding CuSO4 treatment. Inhibition of H+-ATPase activity resulted in reduced Cu accumulation in 30μM CuEDDS-treated roots and 150μM CuEDDS-treated leaves, and induced changes in Cu distribution in the leaves. This indicates that active mechanisms are involved in retaining Cu in the leaf vascular tissues, which prevent its transport to photosynthetically active tissues. The physiological significance of EDDS-assisted Cu uptake is discussed.
► We localised Cu in Brassica carinata treated with CuSO4 or CuEDDS by micro-PIXE. ► EDDS-assisted Cu uptake and transport resulted in preserved root endodermis. ► EDDS enhanced Cu transport from roots to shoots. ► Cu sequestration within leaf veins prevented its mobilisation into the mesophyll. ► EDDS presence conferred tolerance of B. carinata to Cu.
Molecular imaging of cannabis leaf tissue with MeV-SIMS method Jenčič, Boštjan; Jeromel, Luka; Ogrinc Potočnik, Nina ...
Nuclear instruments & methods in physics research. Section B, Beam interactions with materials and atoms,
03/2016, Volume:
371
Journal Article
Peer reviewed
To broaden our analytical capabilities with molecular imaging in addition to the existing elemental imaging with micro-PIXE, a linear Time-Of-Flight mass spectrometer for MeV Secondary Ion Mass ...Spectrometry (MeV-SIMS) was constructed and added to the existing nuclear microprobe at the Jožef Stefan Institute. We measured absolute molecular yields and damage cross-section of reference materials, without significant alteration of the fragile biological samples during the duration of measurements in the mapping mode. We explored the analytical capability of the MeV-SIMS technique for chemical mapping of the plant tissue of medicinal cannabis leaves. A series of hand-cut plant tissue slices were prepared by standard shock-freezing and freeze-drying protocol and deposited on the Si wafer. We show the measured MeV-SIMS spectra showing a series of peaks in the mass area of cannabinoids, as well as their corresponding maps. The indicated molecular distributions at masses of 345.5u and 359.4u may be attributed to the protonated THCA and THCA-C4 acids, and show enhancement in the areas with opened trichome morphology.
Imaging Mass Spectroscopy (IMS) is a unique research tool providing localization and identification of a wide range of biomolecules as essential data to understand biochemical processes in living ...organisms. Secondary Ion Mass Spectrometry with high-energy heavy ions (MeV-SIMS) is emerging as a promising IMS technique for chemical imaging of biological tissue.
We measured the molecular mass spatial distributions in leaves of khat (Catha edulis). Khat is a natural drug plant, native to eastern Africa and the Arabian Peninsula. In these countries, fresh leaves are being chewed by significant part of population. It was reported that 80% of the adult population in Yemen chew the khat leaves. The main stimulating effects of khat are induced by a monoamine alkaloid called cathinone. During leaf ageing, cathinone is further metabolised to cathine and norephedrine. Earlier studies identified the alkaloids in khat, however little is known on their spatial distribution, reflecting the biosynthesis and accumulation in the tissue.
Chemical mapping of alkaloids on cross-sections of khat leaves by MeV-SIMS was done at JSI by a pulsed 5.8MeV 35Cl6+ beam, focused to a diameter of 15μm, using a linear time-of-flight (TOF) spectrometer with a mass resolution of 500. In addition, measurements of MeV-SIMS mass spectra were performed at Kyoto University by a continuous broad beam of 6MeV 63Cu4+ ions at an orthogonal TOF spectrometer with a high mass resolution of 11,000. Sections of leaves were analysed and mass spectra obtained at both MeV-SIMS setups were compared. Tissue-level distributions of detected alkaloids are presented and discussed.
In the last few decades, research has increasingly been aimed at clarifying how root system architecture, physiology and function are related to environmental drivers. âNegative halotropismâ has ...been defined as the alteration of root growth direction to avoid salinity. We suggested that âpositive halotropismâ may be found in halophytes relying on salinity for optimal growth. Investigating root structure of the halophyte Bassia indica (Wight) A. J. Scott, we have shown that positive halotropism can explain the growth of horizontal roots towards optimal salt concentrations along a soil salinity gradient. Here we tested three hypotheses. First, that development of B. indica roots depends on a trade-off between optimal nutrient supply and saline concentrations: results of split-root-experiment showed a preference for sand enriched with nutrients and poor in salts. Second, that shallow horizontal roots enable B. indica to forage for nutrient-rich patches. Results demonstrated that bulk elemental analysis was not consistent with tissue-specific elemental analysis, and this can be explained by substantial variability of element composition of particular root segments. Third, we hypothesised that B. indica redistributes water horizontally through shallow horizontal roots. Results showed that back flow of water from the tap root towards tip root was possible in horizontal roots in saline microenvironment.
Elemental concentrations in micro-PIXE (Particle Induced X-ray Emission) maps of elements in biological tissue slices have been determined using auxiliary information on the sample matrix composition ...from EBS (Elastic Backscattering Spectroscopy) and STIM (Scanning Transmission Ion Microscopy). The thin sample approximation may be used for evaluating micro-PIXE data in cases, where X-ray absorption in the sample can be neglected and the mass of elements in a selected area can be estimated. The resulting sensitivity amounts to an impressive 10−12g of the selected elements. Two cases are presented as examples. In the first, we determined the total mass of gold nanoparticles internalized by human monocyte-derived dendritic cells (MDDC). In the second, an inventory of the mass of elements in the micro-particulate material adsorbed at the wall of the lorica of the microzooplankton species Tintinnopsis radix has been created.
Diverse physiological, biochemical and morphological adaptations enable plants to survive in extreme saline environments where osmotic and ionic stresses limit growth and development. Halophytes are ...salt-tolerant plants that can withstand extraordinarily high levels of Na and Cl in their leaves. The tissue and cellular distribution patterns of salt ions can be linked to the underlying mechanisms of salt tolerance. Application of fast, reliable, multi-elemental and quantitative techniques such as micro-proton-induced X-ray emission (micro-PIXE) will significantly contribute to and accelerate studies of plant salt tolerance, especially as micro-PIXE also provides spatially resolved quantitative data for light elements, such as Na and Mg. The spatial concentration distributions of Na, Mg, Cl, K, P and S in leaves of four halophytes (Bassia indica, Atriplex prostrata, Spartina maritima and Limonium angustifolium) were determined using micro-PIXE, to study the salt-tolerance strategies of the selected halophytes. Different distribution patterns of the studied elements were seen in the leaves; however, in all four of these plant species, Na was excluded from photosynthetically active chlorophyl tissues. With the exception of L. angustifolium, Cl, P and S contents (representing chloride, phosphate and sulphate ionic forms, respectively) did not ensure charge balance in the leaves, which suggests other anionic compounds, such as nitrate and organic anions, have crucial roles in maintaining electroneutrality in these halophytes. By increasing soil salinisation worldwide, the possibility to reliably complement spatial distributions of Na, Mg, Cl, K, P and S with plant structural morphology will contribute significantly to our understanding of plant tolerance mechanisms at the tissue and cell levels. In addition, these kinds of studies are of particular value for designing crop plants with high salt tolerance and for the development of phytoremediation technologies.
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