原子力显微镜 – Page 4 of 13 – Blog • by NanoWorld®

Correlation between plant cell wall stiffening and root extension arrest phenotype in combined abiotic stress of Fe and Al

The plasticity and growth of plant cell walls (CWs) is still not sufficiently understood on its molecular level. *

Atomic Force Microscopy (AFM) has been shown to be a powerful tool to measure the stiffness of plant tissues. *

In the article “Correlation between plant cell wall stiffening and root extension arrest phenotype in the combined abiotic stress of Fe and Al” Harinderbir Kaur, Jean-Marie Teulon, Christian Godon, Thierry Desnos, Shu-wen W. Chen and Jean-Luc Pellequer describe the use of atomic force microscopy (AFM) to observe elastic responses of the root transition zone of 4-day-old Arabidopsis thaliana wild-type and almt1-mutant seedlings grown under Fe or Al stresses. *

In order to evaluate the relationship between root extension and root cell wall elasticity, the authors used Atomic Force Microscopy to perform vertical indentations on surfaces of living plant roots. *

NanoWorld Pyrex-Nitride silicon-nitride PNP-TR AFM probes with triangular AFM cantilevers were used for the nanoindentation experiments with atomic force microscopy. (PNP-TR AFM cantilever beam 2 (CB2) with a typical force constant of 0.08 N/m and a typical resonant frequency of 17 kHz, typical AFM tip radius 10 nm, macroscopic half cone angles 35°). *

Force-distance (F-D) curves were measured using the Atomic Force Microscope and the PNP-TR AFM tips. *

Because of the heterogeneity of seedling CW surfaces, Harinderbir Kaur et al. used the recently developed trimechanics-3PCS framework for interpreting force-distance curves. The trimechanics-3PCS framework allows the extraction of both stiffness and elasticity along the depth of indentation and permits the investigation of the variation of stiffness with varied depth for biomaterials of heterogeneous elasticity responding to an external force. *

A glass slide with a glued seedling (see Figure 1 cited below) was positioned under the AFM cantilever with the help of an AFM optical camera. Due to the large motorized sample stage of the AFM, the glass slide was adjusted in such a way that the AFM cantilever could be positioned perpendicularly at the longitudinal middle of the glued root. The target working area, the transition zone, was 500 µm away from the root apex, almost twice the length of PNP-TR AFM cantilever. *

As shown in the article the presence of single metal species Fe²⁺ or Al³⁺ at 10 μM exerts no noticeable effect on the root growth compared with the control conditions. On the contrary, a mix of both the metal ions produced a strong root-extension arrest concomitant with significant increase of CW stiffness. *

Raising the concentration of either Fe²⁺or Al³⁺ to 20 μM, no root-extension arrest was observed; nevertheless, an increase in root stiffness occurred. In the presence of both the metal ions at 10 μM, root-extension arrest was not observed in the almt1 mutant, which substantially abolishes the ability to exude malate. The authors’ results indicate that the combination of Fe²⁺and Al³⁺ with exuded malate is crucial for both CW stiffening and root-extension arrest. *

It is shown that the elasticity of plant CW is sensitive and can be used to assess abiotic stresses on plant growth and stiffening. *

However, stiffness increase induced by single Fe²⁺ or Al³⁺ is not sufficient for arresting root growth in the described experimental conditions and unexpectedly, the stiffening and the phenotype of seedling roots such as REA are not directly correlated. *

Figure 1 from Harinderbir Kaur et al. 2024 “Correlation between plant cell wall stiffening and root extension arrest phenotype in the combined abiotic stress of Fe and Al”:Principle of nanomechanical measurement of seedling roots with atomic force microscopy. A seedling root (R) is deposited on a microscope slide using silicon glue (N, for Nusil). A fastening band of silicon is seen near the tip of the root (T). The thickness of the fastening band must be thin enough to avoid hindering the AFM support (S), but thick enough to withstand the bending of the root tip. The root is placed under the AFM cantilever (C) as observed by the AFM optical camera. The triangular shaped cantilever (200 µm long) was placed 500 µm away from the root tip in the transition zone where nanoindentation measurements proceeded (as shown). The seedling root and the AFM cantilever are placed within a liquid environment (growth solution, see Supplementary file of the cited article). AFM, atomic force microscopy. NanoWorld Pyrex-Nitride silicon-nitride PNP-TR AFM probes with triangular AFM cantilevers were used for the nanoindentation experiments with atomic force microscopy. — Figure 1 from Harinderbir Kaur et al. 2024 “Correlation between plant cell wall stiffening and root extension arrest phenotype in the combined abiotic stress of Fe and Al”:
Principle of nanomechanical measurement of seedling roots with atomic force microscopy.
A seedling root (R) is deposited on a microscope slide using silicon glue (N, for Nusil). A fastening band of silicon is seen near the tip of the root (T). The thickness of the fastening band must be thin enough to avoid hindering the AFM support (S), but thick enough to withstand the bending of the root tip. The root is placed under the AFM cantilever (C) as observed by the AFM optical camera. The triangular shaped cantilever (200 µm long) was placed 500 µm away from the root tip in the transition zone where nanoindentation measurements proceeded (as shown). The seedling root and the AFM cantilever are placed within a liquid environment (growth solution, see Supplementary file of the cited article). AFM, atomic force microscopy.

*Harinderbir Kaur, Jean‐Marie Teulon, Christian Godon, Thierry Desnos, Shu‐wen W. Chen and Jean‐Luc Pellequer
Correlation between plant cell wall stiffening and root extension arrest phenotype in the combined abiotic stress of Fe and Al
Plant, Cell & Environment 2024; 47:574–584
DOI: https://doi.org/10.1111/pce.14744

The article “Correlation between plant cell wall stiffening and root extension arrest phenotype in the combined abiotic stress of Fe and Al” by Harinderbir Kaur, Jean‐Marie Teulon, Christian Godon, Thierry Desnos, Shu‐wen W. Chen and Jean‐Luc Pellequer is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third-party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.

Electrochemical detection of quinone reduced by Complex I Complex II and Complex III in full mitochondrial membranes

In the last decades enormous advances have been made in characterizing the atomic and molecular structure of respiratory chain supercomplexes. *

However, it still remains a challenge to stitch this refined spatial atomistic description with functional information provided by biochemical studies of isolated protein material. Development of functional assays that detect respiratory chain complexes in their native membrane environment contribute to address the open questions related to the role played by their association and interactions. *

In the article “Electrochemical detection of quinone reduced by Complex I Complex II and Complex III in full mitochondrial membranes” Daniel G. Cava, Julia Alvarez-Malmagro, Paolo Natale, Sandra López-Calcerrada, Iván López-Montero, Cristina Ugalde, Jose Maria Abad, Marcos Pita, Antonio L. De Lacey and Marisela Vélez present a characterization assay in which a functionalized gold electrode is modified with mitochondrial membrane fragments that allows monitoring electrochemically the activity of different respiratory chain complexes immersed in the mitochondrial membrane. *

Daniel G. Cava et al. measure the intensity of the reducing current of the electron mediator CoQ1 at the electrode surface and its variation upon addition of the corresponding enzymatic substrates. The activities of Complex I, Complex II and Complex III were monitored by the way in which they reduce the current, reflecting the amount of quinone reduced by the complexes in the presence of their substrates. *

The authors detect that CoQ1H2 produced by Complex I remains partially trapped within the membrane and is more easily oxidized by Complex III or the electrode than the quinone reduced by Complex II. *

Atomic Force Microscopy (AFM) was used to image the topography of the membrane modified electrode. NanoWorld Pyrex-Nitride Silicon-Nitride AFM probes (PNP-DB, diving board shaped cantilevers, the short AFM cantilever with a typical force constant of 0.48 N/m and 67 kHz resonance frequency) were used. *

The surfaces analysed were the electrodes. The two surfaces imaged are the same previously polished electrodes used for electrochemical measurements. The microscope sample holder was adapted in-home to support the electrodes. Two surfaces were analysed: the polished gold functionalized with 4-aminothiophenol and the electrode after incubation with mitochondria subparticles prepared similarly to the electrodes used for the electrochemical measurements.*

Fig. 2 from Daniel G. Cava et al 2024 “Electrochemical detection of quinone reduced by Complex I Complex II and Complex III in full mitochondrial membranes”
QCM and AFM characterization of modified gold. Panel A shows the frequency (left, black) and dissipation (right red) changes detected on a gold covered quartz crystal previously modified with a 4-ATP after injection in the chamber of the mitochondrial fragments at the time point indicated by the thick arrow. Panel B show AFM images of the surface topography of a modified gold electrode before (left) and after (right)incubation with the mitochondrial membrane. The inset below shows the height profile of the lines indicated in the images.

*Daniel G. Cava, Julia Alvarez-Malmagro, Paolo Natale, Sandra López-Calcerrada, Iván López-Montero, Cristina Ugalde, Jose Maria Abad, Marcos Pita, Antonio L. De Lacey and Marisela Vélez
Electrochemical detection of quinone reduced by Complex I Complex II and Complex III in full mitochondrial membranes
Electrochimica Acta, Volume 484, 20 April 2024, 144042
DOI: https://doi.org/10.1016/j.electacta.2024.144042

The article “Electrochemical detection of quinone reduced by Complex I Complex II and Complex III in full mitochondrial membranes” by Daniel G. Cava, Julia Alvarez-Malmagro, Paolo Natale, Sandra López-Calcerrada, Iván López-Montero, Cristina Ugalde, Jose Maria Abad, Marcos Pita, Antonio L. De Lacey and Marisela Vélez is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third-party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.

Studying the cation dependence of CO2 reduction intermediates at Cu by in situ VSFG spectroscopy

The nature of the electrolyte cation is known to have a significant impact on electrochemical reduction of CO2 at catalyst|electrolyte interfaces. An understanding of the underlying mechanism responsible for catalytic enhancement as the alkali metal cation group is descended is key to guide catalyst development. *

In the article “Studying the cation dependence of CO2 reduction intermediates at Cu by in situ VSFG spectroscopy” Liam C. Banerji, Hansaem Jang, Adrian M. Gardner and Alexander J. Cowan use in situ vibrational sum frequency generation (VSFG) spectroscopy to monitor changes in the binding modes of the CO intermediate at the electrochemical interface of a polycrystalline Cu electrode during CO2 reduction as the electrolyte cation is varied. *

Three alkali metal cations have been chosen for analysis: K+, which is the most commonly used electrolyte cation for eCO2R, Cs+, which has been shown to give the greatest enhancement for C2+ products, and Na+, which shows poorer eCO2R performance than K+ whilst maintaining appreciable levels of C-based products. The ability of VSFG to study catalyst|electrolyte interfaces without the need for modifications, as required in the spectroelectrochemical studies mentioned in the article, which can fundamentally alter the electrodes activity, makes it an important tool to assess the mechanisms occurring on the pc-Cu electrodes routinely employed for eCO2R. *

A CObridge mode is observed only when using Cs+, a cation that is known to facilitate CO2 reduction on Cu, supporting the proposed involvement of CObridge sites in CO coupling mechanisms during CO2 reduction. Ex situ measurements show that the cation dependent CObridge modes correlate with morphological changes of the Cu surface. *

The results presented in the article suggest that a high level of bridge site formation is related to, or facilitated by, the Cu restructuring that happens as a result of the use of the Cs+ cations in the supporting electrolyte. Recent reports have indicated that multiple (bridge) bound CO may be electrochemically inert but this work builds on the emerging evidence that CObridge sites are a key intermediate in the CO–CO coupling step that is required for C2+ formation during eCO2R. *

NanoWorld Pointprobe® CONTR AFM probes for contact mode atomic force microscopy (AFM) were used to characterize the morphology of the CU electrode surface before bulk electrolysis and after bulk electrolysis.*

Fig. 5 from Liam C. Banerji et al. “Studying the cation dependence of CO2 reduction intermediates at Cu by in situ VSFG spectroscopy”: AFM images showing surface morphology of the Cu electrode surface (a) before bulk electrolysis, after bulk electrolysis in CO2 purged 0.5 M (b) NaHCO3, (c) KHCO3 and (d) CsHCO3 and also in (e) CO purged 0.5 M CsHCO3. Image analysis methods are described in the Experimental section.NanoWorld Pointprobe® CONTR AFM probes for contact mode atomic force microscopy (AFM) were used to characterize the morphology of the CU electrode surface before bulk electrolysis and after bulk electrolysis. — Fig. 5 from Liam C. Banerji et al. “Studying the cation dependence of CO2 reduction intermediates at Cu by in situ VSFG spectroscopy”: AFM images showing surface morphology of the Cu electrode surface (a) before bulk electrolysis, after bulk electrolysis in CO2 purged 0.5 M (b) NaHCO3, (c) KHCO3 and (d) CsHCO3 and also in (e) CO purged 0.5 M CsHCO3. Image analysis methods are described in the Experimental section of the original article.

*Liam C. Banerji, Hansaem Jang, Adrian M. Gardner and Alexander J. Cowan
Studying the cation dependence of CO2 reduction intermediates at Cu by in situ VSFG spectroscopy
Chemical Science 2024, 15, 2889-2897
DOI: https://doi.org/10.1039/D3SC05295H

The article “Studying the cation dependence of CO2 reduction intermediates at Cu by in situ VSFG spectroscopy” by Liam C. Banerji, Hansaem Jang, Adrian M. Gardner and Alexander J. Cowan is licensed under a Creative Commons Attribution 3.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third-party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/3.0/.