Protocols in Current Issue
    Quantification of Plant Cell Death by Electrolyte Leakage Assay
    Authors:  Noriyuki Hatsugai and Fumiaki Katagiri, date: 03/05/2018, view: 5442, Q&A: 1
    [Abstract] We describe a protocol to measure the electrolyte leakage from plant tissues, resulting from loss of cell membrane integrity, which is a common definition of cell death. This simple protocol is designed to measure the electrolyte leakage from a tissue sample over a time course, so that the extent of cell death in the tissue can be monitored ...
    Isolation and Quantification of Plant Extracellular Vesicles
    Authors:  Brian D. Rutter, Katie L. Rutter and Roger W. Innes, date: 09/05/2017, view: 7343, Q&A: 3
    [Abstract] Extracellular vesicles (EVs) play an important role in intercellular communication by transporting proteins and RNA. While plant cells secrete EVs, they have only recently been isolated and questions regarding their biogenesis, release, uptake and function remain unanswered. Here, we present a detailed protocol for isolating EVs from the ...
    Quantification of Low Molecular Weight Thiols in Arabidopsis
    Authors:  Ziqing Miao, Zhen Wang and Cheng-Bin Xiang, date: 01/05/2016, view: 4246, Q&A: 0
    [Abstract] Low-molecular-weight (LMW) thiols are a class of highly reactive compounds due to their thiol moiety. They play important roles in the maintenance of cellular redox homeostasis, detoxification, and development. Monobromobimane (mBBr) is weakly fluorescent but selectively reacts with thiols to yield highly fluorescent thioethers (mBSR) products, ...
    Luminol-based Assay for Detection of Immunity Elicitor-induced Hydrogen Peroxide Production in Arabidopsis thaliana Leaves
    Authors:  Nora Gigli Bisceglia, Matteo Gravino and Daniel V. Savatin, date: 12/20/2015, view: 9062, Q&A: 1
    [Abstract] In Arabidopsis thaliana, one of the very early immune-related responses induced after elicitor perception is the oxidative burst, i.e., reactive oxygen species (ROS) generation including superoxide anion and hydrogen peroxide (H2O2). ROS production plays different roles in a wide range of biotic and abiotic ...
    Extraction of Apoplastic Wash Fluids and Leaf Petiole Exudates from Leaves of Arabidopsis thaliana
    Authors:  Takao Araya, Anne Bohner and Nicolaus von Wirén, date: 12/20/2015, view: 7613, Q&A: 0
    [Abstract] The long-distance translocation of metabolites and mineral elements is crucial for plant growth and reproduction. In most cases, source-to-sink translocation of metabolites and minerals requires their passage through the apoplast, irrespective whether they are transported via the xylem or the phloem. This apoplast-mediated pathway is of particular ...
    Quantification of Callose Deposition in Plant Leaves
    [Abstract] Callose is an amorphous homopolymer, composed of β-1, 3-glucan, which is widespread in higher plants. Callose is involved in multiple aspects of plant growth and development. It is synthetized in plants at the cell plate during cytokinesis, in several stages during pollen development and is deposited at plasmodesmata to regulate the cell-to-cell ...
    High-throughput Quantification of Ammonium Content in Arabidopsis
    Authors:  Izargi Vega-Mas, Asier Sarasketa and Daniel Marino, date: 08/20/2015, view: 5629, Q&A: 0
    [Abstract] This protocol is a simple colorimetric assay for internal ammonium quantification in aqueous extracts from plant tissues. The method is based on the phenol hypochlorite assay (Berthelot reaction):

    NH4+ + hypochlorite + OH- + phenol → indophenol

    The oxidation of indophenol caused by phenol ...
    Detection of Alternative Oxidase Expression in Arabidopsis thaliana Protoplasts Treated with Aluminium
    Authors:  Jian Liu, Zhe Li and Da Xing, date: 07/05/2015, view: 5076, Q&A: 0
    [Abstract] Aluminium (Al), a non-essential metal widespread in the environment that is known to be toxic to humans as well as to plants, can cause damage not only to the roots but also to the aerial parts of plants. Its toxicity has been recognized as one of the major factors that limit crop production on acid soil. Alternative oxidase, the respiratory ...
    Chlorophyll Fluorescence Measurements in Arabidopsis Plants Using a Pulse-amplitude-modulated (PAM) Fluorometer
    Authors:  Reiko Motohashi and Fumiyoshi Myouga, date: 05/05/2015, view: 10586, Q&A: 0
    [Abstract] In this protocol, to analyze PSII activity in photosynthesis, we measure the Fv/Fm (Fv=Fm ± Fo) value (Fo and Fm are the minimum and maximum values of chlorophyll fluorescence of dark-adapted leaves, respectively). Fv/Fm is a reliable marker of photo- inhibition (Krause et al., 1988). Chlorophyll fluorescence in leaves was measured at ...
    Camalexin Quantification in Arabidopsis thaliana Leaves Infected with Botrytis cinerea
    [Abstract] Phytoalexins are heterogeneous low molecular mass secondary metabolites with antimicrobial activity produced at the infection site in response to pathogen invasion and represent an important part of the plant defense repertoire. Camalexin (3-Thiazol-2′-yl-indole) is a known phytoalexin first detected and isolated in Camelina sativa, from ...

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