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    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: 84, 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 ...
    Analysis of Chromosome Condensation/Decondensation During Mitosis by EdU Incorporation in Nigella damascena L. Seedling Roots
    Author:  Eugene V. Sheval, date: 02/05/2018, view: 90, Q&A: 0
    [Abstract] To investigate the chromosome dynamics during mitosis, it is convenient to mark the discrete chromosome foci and then analyze their spatial rearrangements during prophase condensation and telophase decondensation. To label the chromosome regions in plant chromosomes, we incorporated the synthetic nucleotide, 5-ethynyl-2’-deoxyuridine (EdU), which ...
    Electron Tomography to Study the Three-dimensional Structure of Plasmodesmata in Plant Tissues–from High Pressure Freezing Preparation to Ultrathin Section Collection
    Authors:  William J. Nicolas, Emmanuelle Bayer and Lysiane Brocard, date: 01/05/2018, view: 377, Q&A: 0
    [Abstract] Plasmodesmata (PD) are nanometric (~20 nm wide) membrane lined pores encased in the cell walls of the adjacent plant cells. They allow the cells to exchange all types of molecules ranging from nutrients like sugar, hormones, to RNAs and various proteins. Unfortunately, they are also hijacked by phyto-viruses, enabling them to spread from ...
    Biomechanical Characterization of Onion Epidermal Cell Walls
    Authors:  Daniel M. Durachko, Yong Bum Park, Tian Zhang and Daniel J. Cosgrove, date: 12/20/2017, view: 443, Q&A: 1
    [Abstract] Here we describe two experimental protocols to measure the biomechanical properties of primary (growing) plant cell walls, with a focus on analyzing cell wall epidermal strips of onion scales. The first protocol measures cell wall creep (time-dependent irreversible extension) under constant force. Such creep is often mediated by the wall-loosening ...
    Preparation of Onion Epidermal Cell Walls for Imaging by Atomic Force Microscopy (AFM)
    Authors:  Tian Zhang and Daniel J. Cosgrove, date: 12/20/2017, view: 616, Q&A: 0
    [Abstract] The growing plant cell wall is comprised of long, thin cellulose microfibrils embedded in a hydrated matrix of polysaccharides and glycoproteins. These components are typically constructed in layers (lamellae) on the inner surface of the cell wall, i.e., between the existing wall and the plasma membrane. The organization of these ...
    Determination of H+-ATPase Activity in Arabidopsis Guard Cell Protoplasts through H+-pumping Measurement and H+-ATPase Quantification
    Authors:  Shota Yamauchi and Ken-ichiro Shimazaki, date: 12/20/2017, view: 522, Q&A: 0
    [Abstract] The opening of stomata in plants in response to blue light is driven by the plasma membrane H+-ATPase in guard cells. To evaluate the activation of the H+-ATPase in vivo, we can use H+-pumping by guard cells in response to blue light and fusicoccin. To do this, it is required to prepare a large amount of ...
    Automatic Leaf Epidermis Assessment Using Fourier Descriptors in Texture Images
    [Abstract] The identification of plant species is not a trivial task, since it is carried out by experts and depends on the presence of fruits, flowers and leaves. However, fruits and flowers are not available throughout the year, while leaves are accessible most of the year. In order to assist the specialized work of species identification, methods of ...
    Using Light and Electron Microscopy to Estimate Structural Variation in Thylakoid Membranes
    Authors:  Hatsumi Nozue and Kiyokazu Kametani, date: 12/05/2017, view: 504, Q&A: 0
    [Abstract] The shapes of chloroplasts and the architectures of internal thylakoid membranes are altered by growth and environmental changes (Lichtenthaler et al., 1981; Kutik, 1985; Terashima and Hikosaka, 1995). These morphological alterations proceed via transitional intermediates, during which dynamic and heterogeneous thylakoid membranes are ...
    [Bio101] MTT Assay for Cytotoxicity Assessment in Oryza sativa Root Tissue
    Authors:  Snehalata Majumdar, Titir Guha and Rita Kundu, date: 11/20/2017, view: 609, Q&A: 0
    [Abstract] Cytotoxicity of different compounds are commonly evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. This assay is mainly used to study cell viability in cell lines (Carmichael et al., 1987). In this study, the protocol is being used to determine the cell viability of plant roots, treated with different ...
    Histochemical Preparations to Depict the Structure of Cauliflower Leaf Hydathodes
    Authors:  Aude Cerutti, Marie-Christine Auriac, Laurent D. Noël and Alain Jauneau, date: 10/20/2017, view: 1281, Q&A: 0
    [Abstract] Hydathodes are plant organs present on leaf margins of a wide range of vascular plants and are the sites of guttation. Both anatomy and physiology of hydathodes are poorly documented. We have recently reported on the anatomy of cauliflower and Arabidopsis thaliana hydathodes and on their infection by the vascular pathogenic bacterium ...