Sequential quantitative X‐ray elemental imaging of frozen‐hydrated and freeze‐dried biological bulk samples in the SEM

Marshall, A.T.; Goodyear, Melinda

doi:10.1111/j.1365-2818.2011.03539.x

Cited by 12 publications

(11 citation statements)

References 26 publications

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“…Summed spectra from regions of interest were quantified using the AZtec model for matrix corrections (Marshall 2017). This sample preparation method and analytical system have been shown to be highly suitable for cellular element analyses of biological samples (Huang et al 1994, Marshall and Xu 1998, Marshall and Clode 2009, McCully et al 2010, Marshall et al 2012, Marshall 2017, Ding et al 2018, Guilherme Pereira et al 2018, Hayes et al 2018.…”

Section: Distribution Of P and Ca Within The Leaf Cellmentioning

confidence: 99%

Effects of calcium and its interaction with phosphorus on the nutrient status and growth of three Lupinus species

Ding

Clode

Clements

et al. 2018

Physiologia Plantarum

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Phosphorus (P)-deficiency symptoms are known for Lupinus species grown in calcareous soil, but we do not know if this is due to a high calcium (Ca) availability or a low P availability in the soil. To address this problem, we explored both the effects of Ca and its interactions with P on nutrient status and growth of three Lupinus species. Two Ca-sensitive genotypes (L. angustifolius L. P26723 and L. cosentinii Guss. P27225) and two Ca-tolerant genotypes (L. angustifolius L. cv Mandelup and L. pilosus Murr. P27440) were grown hydroponically at two P (0.1 and 10 μM) and three Ca (0.1, 0.6 and 6 mM) levels. Leaf symptoms and biomass were recorded, whole leaf and root nutrient concentrations were analysed, and leaf cellular P and Ca concentrations were determined. Phosphorus-deficiency symptoms were only observed in the Ca-sensitive genotypes. Among all the genotypes in this study, the Ca-tolerant L. pilosus showed an ability to maintain stable leaf Ca and P concentrations whereas the Ca-tolerant L. angustifolius cv Mandelup did not maintain a stable whole leaf Ca concentration, but maintained a low cytosolic Ca concentration through effective Ca compartmentation. However, the two Ca-sensitive genotypes, L. angustifolius P26723 and L. cosentinii, did not exhibit an ability to maintain a stable whole leaf Ca concentration or effectively compartmentalise Ca. Therefore, having the capacity to maintain a stable whole leaf Ca concentration or effectively compartmentalising Ca in leaves are likely critical for Lupinus species to be Ca tolerant.

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Section: Distribution Of P and Ca Within The Leaf Cellmentioning

confidence: 99%

Effects of calcium and its interaction with phosphorus on the nutrient status and growth of three Lupinus species

Ding

Clode

Clements

et al. 2018

Physiologia Plantarum

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“…Moreover, the mean values of elemental concentrations derived from an X-ray image represent several hundreds to thousands of probe measurements, which provide more reliable information than dozens of randomly selected measurement points. Previous studies have described results from frozen-hydrated biological materials analyzed with X-ray imaging in a SEM system [9,10]. However, quantitative elemental imaging in a TEM system has seldom been attempted in biological materials.…”

Section: Introductionmentioning

confidence: 99%

Quantitative X-ray Elemental Imaging in Plant Materials at the Subcellular Level with a Transmission Electron Microscope: Applications and Limitations

et al. 2014

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Energy-dispersive X-ray microanalysis (EDX) is a technique for determining the distribution of elements in various materials. Here, we report a protocol for high-spatial-resolution X-ray elemental imaging and quantification in plant tissues at subcellular levels with a scanning transmission electron microscope (STEM). Calibration standards were established by producing agar blocks loaded with increasing KCl or NaCl concentrations. TEM-EDX images showed that the salts were evenly distributed in the agar matrix, but tended to aggregate at high concentrations. The mean intensities of K+, Cl−, and Na+ derived from elemental images were linearly correlated to the concentrations of these elements in the agar, over the entire concentration range tested (R > 0.916). We applied this method to plant root tissues. X-ray images were acquired at an actual resolution of 50 nm × 50 nm to 100 nm × 100 nm. We found that cell walls exhibited higher elemental concentrations than vacuoles. Plants exposed to salt stress showed dramatic accumulation of Na+ and Cl− in the transport tissues, and reached levels similar to those applied in the external solution (300 mM). The advantage of TEM-EDX mapping was the high-spatial-resolution achieved for imaging elemental distributions in a particular area with simultaneous quantitative analyses of multiple target elements.

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“…After coating, samples were transferred under vacuum to a Zeiss field emission scanning electron microscope fitted with a Leica VCT100 cryotransfer and stage, and an Oxford X‐Max80 SDD X‐ray detector interfaced to Oxford Instruments AZtecEnergy software (Figure S1b). This preparation method and this fully integrated Oxford analytical system have been shown to be highly suitable for elemental analysis and quantitation of biological samples (Huang, Canny, Oates, & McCully, ; Jin et al, ; Marshall, ; Marshall & Clode, ; Marshall, Goodyear, & Crewther, ; Marshall & Xu, ; McCully, Canny, Huang, Miller, & Brink, ).…”

Section: Methodsmentioning

confidence: 99%

“…Oxford analytical system have been shown to be highly suitable for elemental analysis and quantitation of biological samples (Huang, Canny, Oates, & McCully, 1994;Jin et al, 2017;Marshall, 2017;Marshall & Clode, 2009;Marshall, Goodyear, & Crewther, 2012;Marshall & Xu, 1998;McCully, Canny, Huang, Miller, & Brink, 2010).…”

Section: Cell-specific Element Analysis By X-ray Microanalytical Mamentioning

confidence: 99%

Proteaceae from phosphorus‐impoverished habitats preferentially allocate phosphorus to photosynthetic cells: An adaptation improving phosphorus‐use efficiency

Hayes

Clode

Oliveira

et al. 2018

Plant Cell & Environment

103

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Plants allocate nutrients to specific leaf cell types; eudicots are thought to predominantly allocate phosphorus (P) to epidermal/bundle sheath cells. However, three Proteaceae species have been shown to preferentially allocate P to mesophyll cells instead. These Proteaceae species are highly adapted to P-impoverished habitats, with exceptionally high photosynthetic P-use efficiencies (PPUE). We hypothesized that preferential allocation of P to photosynthetic mesophyll cells is an important trait in species adapted to extremely P-impoverished habitats, contributing to their high PPUE. We used elemental X-ray mapping to determine leaf cell-specific nutrient concentrations for 12 Proteaceae species, from habitats of strongly contrasting soil P concentrations, in Australia, Brazil, and Chile. We found that only species from extremely P-impoverished habitats preferentially allocated P to photosynthetic mesophyll cells, suggesting it has evolved as an adaptation to their extremely P-impoverished habitat and that it is not a family-wide trait. Our results highlight the possible role of soil P in driving the evolution of ecologically relevant nutrient allocation patterns and that these patterns cannot be generalized across families. Furthermore, preferential allocation of P to photosynthetic cells may provide new and exciting strategies to improve PPUE in crop species.

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Sequential quantitative X‐ray elemental imaging of frozen‐hydrated and freeze‐dried biological bulk samples in the SEM

Cited by 12 publications

References 26 publications

Effects of calcium and its interaction with phosphorus on the nutrient status and growth of three Lupinus species

Effects of calcium and its interaction with phosphorus on the nutrient status and growth of three Lupinus species

Quantitative X-ray Elemental Imaging in Plant Materials at the Subcellular Level with a Transmission Electron Microscope: Applications and Limitations

Proteaceae from phosphorus‐impoverished habitats preferentially allocate phosphorus to photosynthetic cells: An adaptation improving phosphorus‐use efficiency

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