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Visualization of Plant Cell Wall Epitopes Using Immunogold Labeling for Electron Microscopy

Author: Mateusz Majda, date: 04/05/2019, view: 330, Q&A: 0

[Abstract] Plant cell walls consist of different polysaccharides and structural proteins, which form a rigid layer located outside of the plasma membrane. The wall is also a very dynamic cell composite, which is characterized by complex polysaccharide interactions and various modifications during cell development. The visualization of cell wall components in ...

Quantitative Plasmodesmata Permeability Assay for Pavement Cells of Arabidopsis Leaves

Authors: Min Diao, Qiannan Wang and Shanjin Huang, date: 04/05/2019, view: 318, Q&A: 0

[Abstract] Plasmodesmata (PD) are intercellular channels between walled plant cells that enable the transportation of materials between adjacent cells, which are important for plant growth and development. The permeability of PD must be tightly regulated. Assays to determine the permeability of PD are crucial for related studies on the regulation of PD ...

Visualization and Quantification of Cell-to-cell Movement of Proteins in Nicotiana benthamiana

Authors: Mila C Blekemolen, Mariliis Tark-Dame and Frank L.W Takken, date: 12/20/2018, view: 1044, Q&A: 0

[Abstract] Cell-to-cell movement of proteins through plasmodesmata is a widely-established mechanism for intercellular signaling in plants. Current techniques to study intercellular protein translocation rely on single-cell transformation using particle bombardment or transgenic lines expressing photo-inducible fluorophores. The method presented here allows ...

Using Arabidopsis Mesophyll Protoplasts to Study Unfolded Protein Response Signaling

Authors: Yan Bao and Diane C. Bassham, date: 12/05/2018, view: 939, Q&A: 0

[Abstract] Various environmental stresses or artificial reagents can trigger unfolded protein accumulation in the endoplasmic reticulum (ER) due to the folding capacity of the ER being exceeded. This is termed ER stress, and triggers the unfolded protein response (UPR). Assays for activation of the UPR in plants include Tunicamycin (Tm)- or dithiothreitol ...

An Image Analysis Pipeline to Quantify Emerging Cracks in Materials or Adhesion Defects in Living Tissues

Authors: Stéphane Verger, Guillaume Cerutti and Olivier Hamant, date: 10/05/2018, view: 988, Q&A: 0

[Abstract] Microcracks in materials reflect their mechanical properties. The quantification of the number or orientation of such cracks is thus essential in many fields, including engineering and geology. In biology, cracks in soft tissues can reflect adhesion defects, and the analysis of their pattern can help to deduce the magnitude and orientation of ...

Artificial Inoculation of Epichloë festucae into Lolium perenne, and Visualisation of Endophytic and Epiphyllous Fungal Growth

Authors: Yvonne Becker, Kimberly A. Green, Barry Scott and Matthias Becker, date: 09/05/2018, view: 1288, Q&A: 0

[Abstract] Natural hosts for the fungal endophyte Epichloë festucae include Festuca rubra (fine fescue) and Festuca trachyphylla (hard fescue). Some strains also form stable associations with Lolium perenne (perennial ryegrass). L. perenne is a suitable host to study fungal endophyte–grass interactions, such as endophytic ...

Increasing the Membrane Permeability of a Fern with DMSO

Author: Marcelo Garcés, date: 06/20/2018, view: 1353, Q&A: 0

[Abstract] Cell membrane prevents the entrance of extra molecules (e.g., transcription and translation inhibitors) into the cell. For studying the physiological effects of transcription and translation inhibitors on Hymenophyllum caudiculatum fronds, we incubate fronds with 0.1% DMSO to test if this increases cell membrane permeability ...

Slow and Fast Desiccation of Single-cell Thick Fronds of Filmy Ferns

Author: Marcelo Garcés, date: 06/05/2018, view: 1119, Q&A: 0

[Abstract] Filmy ferns can desiccate and recover after rehydration to resume photosynthesis. Slow and fast desiccation rates were compared in filmy fern fronds to determine whether structural or physiological differences may occur between desiccation rates. Slow desiccation is considered to be more similar to natural conditions experienced by plants that ...

Fluorescein Transport Assay to Assess Bulk Flow of Molecules Through the Hypocotyl in Arabidopsis thaliana

Authors: Salva Duran-Nebreda and George W. Bassel, date: 04/05/2018, view: 1732, Q&A: 0

[Abstract] The bulk transport of molecules through plant tissues underpins growth and development. The stem acts as a conduit between the upper and low domains of the plant, facilitating transport of solutes and water from the roots to the shoot system, and sugar plus other elaborated metabolites towards the non-photosynthetic organs. In order to perform ...

Investigating Localization of Chimeric Transporter Proteins within Chloroplasts of Arabidopsis thaliana

Authors: Susumu Uehara, Yasuko Ito-Inaba and Takehito Inaba, date: 02/05/2018, view: 2636, Q&A: 0

[Abstract] In this protocol, we describe a method to design chimeric proteins for specific targeting to the inner envelope membrane (IEM) of Arabidopsis chloroplasts and the confirmation of their localization by biochemical analysis. Specific targeting to the chloroplast IEM can be achieved by fusing the protein of interest with a transit peptide ...

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