The Early Entry of Al into Cells of Intact Soybean Roots (A Comparison of Three Developmental Root Regions Using Secondary Ion Mass Spectrometry Imaging)

Lazof, Dennis B.; Goldsmith, Jack G.; Rufty, Thomas W.; Linton, Richard W.

doi:10.1104/pp.112.3.1289

Cited by 46 publications

(24 citation statements)

References 35 publications

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“…Linton and Goldsmith 1992;Lazof et al 1996;Grignon et al 1996;Derue et al 2006). The choice for a particular method of sample preparation is problem-and sample typedependent.…”

Section: Passive Vs Active Mechanisms Of Ge-si Fractionationmentioning

confidence: 99%

Subcellular localization of silicon and germanium in grass root and leaf tissues by SIMS: evidence for differential and active transport

et al. 2010

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Silicon transport and incorporation into plant tissue is important to both plant physiological function and to the influence plants have on ecosystem silica cycling. However, the mechanisms controlling this transport have only begun to be explored. In this study, we used secondary ion mass spectrometry (SIMS) to image concentrations of Si in root and shoot tissues of annual blue grass (Poa annua L.) and orchard grass (Dactylis glomerata L.) with the goal of identifying control points in the plant silica uptake pathway. In addition, we used SIMS to describe the distributions of germanium (Ge); the element used to trace Si in biogeochemical studies. Within root tissue, Si and Ge were localized in the suberized thickwalled region of endodermal cells, i.e. the proximal side of endodermal cells which is in close association to the casparian strip. In leaves, Si was present in the cell walls, but Ge was barely detectable. The selective localization of Si and Ge in the proximal side of endodermal cell walls of roots suggests transport control is exerted upon Si and Ge by the plant. The absence of Si in most root cell walls and its presence in the cell walls of leaves (in areas outside of the transpiration terminus) suggests modifications in the chemical form of Si to a form that favors Si complexation in the cell walls of leaf tissue. The low abundance of Ge in leaf tissue is consistent with previous studies that suggest preferential transport of Si relative to Ge.

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“…Linton and Goldsmith 1992;Lazof et al 1996;Grignon et al 1996;Derue et al 2006). The choice for a particular method of sample preparation is problem-and sample typedependent.…”

Section: Passive Vs Active Mechanisms Of Ge-si Fractionationmentioning

confidence: 99%

Subcellular localization of silicon and germanium in grass root and leaf tissues by SIMS: evidence for differential and active transport

et al. 2010

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“…To complement this research, microanalytical studies using appropriate techniques should measure short‐term uptake and exchange kinetics of metal ions within the apoplast of mycorrhizas in comparison to non‐mycorrhizal roots with special regards to meristematic regions. Although intracellular uptake across the plasma membrane is difficult to estimate, secondary ion mass spectrometry (SIMS) has the potential to do so ( Lazof et al 1996). This could show whether, or at what structural level, mycorrhizas may exclude metals from uptake.…”

Section: Conclusion and Future Research Needsmentioning

confidence: 99%

Metal toxicity and ectomycorrhizas

Jentschke

Godbold

2000

Physiologia Plantarum

251

136

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Metal toxicity (Al and heavy metals) is a major constraint affecting root growth in a number of natural or managed ecosystems. Fine roots of the majority of plant species are associated with mycorrhizal fungi, which may modify the sensitivity of roots to metal stress. In this review, we summarise the available evidence demonstrating beneficial effects of ectomycorrhizas in alleviation of metal toxicity in forest tree seedlings. We identify experimental shortcomings of past research (e.g. the use of shoot metal concentrations as a measure of metal uptake, use of microanalytical techniques biased by element redistribution) that may confound major conclusions drawn from these experiments. Although there is no doubt that in many cases ectomycorrhizal fungi indeed ameliorate metal stress in their host plants, the mechanism(s) involved remain(s) unclear. The role of metal sorption on fungal tissues thought to reduce metal exposure of the host plant is critically reviewed. As direct evidence (both under artificial and soil conditions) supporting a unique role of fungal immobilisation of metals is lacking so far, there is an urgent need to also test alternative tolerance mechanisms such as the release of metal chelating substances, or nutritional and hormonal effects mediated by mycorrhizal fungi.

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“…Cryopreparations are required for analysis of substances, which are potentially diffusible, although these preparations lower spatial resolution and lack structural detail (Echlin, 1991;Lazof et al, 1996). The objective of the fixation procedure is to maintain the in vivo location of elemental ions or small molecules.…”

Section: Discussionmentioning

confidence: 99%

Preparation of cells cultured on silicon wafers for mass spectrometry analysis

Wittig

Wiemann

Fartmann

et al. 2005

Microscopy Res & Technique

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The distribution of specific atoms and molecules within living cells is of high interest in bio-medical research. Laser secondary neutral mass spectrometry (laser-SNMS) and time-of-flight secondary ion mass spectrometry (TOF-SIMS) detect atoms with high sensitivity and spatial resolution. The application of these methods to cultured cells requires special preparation techniques preserving morphological and chemical integrity of the living cells. The cells should, therefore, be grown on a conducting material preventing charging of the sample during ion bombardment. Silicon is currently used as the preferred support material for non-biological samples in mass spectrometry. This study investigates (1) the influence of silicon surfaces on cell growth and (2) the suitability of a sandwiched, rapid freezing method to analyse transmembrane ion gradients. Human melanoma cells were grown on silicon with polished or etched surfaces. Growth kinetics were studied using the Sulforhodamine-B assay. Number, shape, and morphology of the cells were assessed by epifluorescence microscopy of calcein AM- and DAPI-stained cells. Cells were subjected to rapid freezing, freeze-fracturing, and freeze-drying prior to analysis by TOF-SIMS and laser-SNMS. While cell numbers and morphology on the rough silicon wafers were impaired, morphology and growth kinetics of cells on polished silicon were identical to control cells on cell culture tested polystyrene. TOF-SIMS and laser-SNMS resulted in high-resolution elemental images and mass spectra. Measurement of the intracellular Na+ and K+ concentrations revealed a ratio as observed in living cells. In conclusion, culturing cells on polished silicon wafers followed by sandwiched, rapid freezing is an adequate preparation method to study intracellular ion distribution with mass spectrometry.

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The Early Entry of Al into Cells of Intact Soybean Roots (A Comparison of Three Developmental Root Regions Using Secondary Ion Mass Spectrometry Imaging)

Cited by 46 publications

References 35 publications

Subcellular localization of silicon and germanium in grass root and leaf tissues by SIMS: evidence for differential and active transport

Subcellular localization of silicon and germanium in grass root and leaf tissues by SIMS: evidence for differential and active transport

Metal toxicity and ectomycorrhizas

Preparation of cells cultured on silicon wafers for mass spectrometry analysis

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