Plants

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    Protocols in Current Issue
    Low-cost and High-throughput RNA-seq Library Preparation for Illumina Sequencing from Plant Tissue
    Authors:  Marta Bjornson, Kaisa Kajala, Cyril Zipfel and Pingtao Ding, date: 10/20/2020, view: 702, Q&A: 0
    [Abstract] Transcriptome analysis can provide clues to biological processes affected in different genetic backgrounds or/and under various conditions. The price of RNA sequencing (RNA-seq) has decreased enough so that medium- to large-scale transcriptome analyses in a range of conditions are feasible. However, the price and variety of options for library ...
    Expression and Purification of Arabidopsis Transmembrane Protein BCM1 in Saccharomyces cerevisiae
    Authors:  Peng Wang and Bernhard Grimm, date: 09/20/2020, view: 621, Q&A: 0
    [Abstract] Heterologous expression and purification of transmembrane proteins have remained a challenge for decades hampering detailed biochemical and structural characterization of key enzymes and their interacting regulators in multiple metabolic pathways. An in-depth study on the newly identified Arabidopsis thaliana integral membrane protein ...
    The ATPase Activity of Escherichia coli Expressed AAA+-ATPase Protein
    Authors:  Amita Kaundal, Vemanna S. Ramu and Kirankumar S. Mysore, date: 08/05/2020, view: 911, Q&A: 0
    [Abstract] ATPases are the enzymes that breakdown ATP to ADP and release inorganic phosphate (Pi). Here we provide a detailed protocol to determine the ATPase activity of a recombinant AAA+-ATPase protein (GENERAL CONTROL NON-REPRESSIBLE-4 [GCN4]) by spectrophotometric absorption at 360 nm to measure the accumulated inorganic phosphate. In ...
    Maintenance and Quantitative Phenotyping of the Oomycete-plant Model Pathosystem Hyaloperonospora arabidopsidisArabidopsis
    Authors:  Fang-Yu Hwu and Martin Parniske, date: 06/20/2020, view: 1025, Q&A: 0
    [Abstract] The interaction between the host plant Arabidopsis thaliana (Arabidopsis) and the oomycete Hyaloperonospora arabidopsidis (Hpa) is an established model system for the study of an obligate biotrophic downy mildew interaction. The evaluation of the developmental success of Hpa is often based on the quantification ...
    Measurement of Chloroplastic NAD Kinase Activity and Whole Tissue NAD Kinase Assay
    Authors:  Yuuma Ishikawa, Maki Kawai-Yamada and Shin-nosuke Hashida, date: 01/05/2020, view: 1788, Q&A: 0
    [Abstract] Nicotinamide adenine dinucleotide phosphate (NADP) synthesis requires nicotinamide adenine dinucleotide (NAD) kinase activity, substrate NAD and ATP. The NAD kinase responds to various environmental stimuli and its activity is regulated via various regulatory pathways, such as Ca2+-dependent and redox-dependent signals. Conventional in ...
    Method for Assessing Virulence of Colletotrichum higginsianum on Arabidopsis thaliana Leaves Using Automated Lesion Area Detection and Measurement
    Authors:  Ayako Tsushima, Pamela Gan and Ken Shirasu, date: 11/20/2019, view: 2733, Q&A: 0
    [Abstract] The plant pathogenic fungus, Colletotrichum higginsianum is widely used to understand infection mechanisms, as it infects the model plant Arabidopsis thaliana. To determine the virulence of C. higginsianum, several methods have been developed, such as disease reaction scoring, lesion measurement, entry rate assays, and ...
    Non-aqueous Fractionation (NAF) for Metabolite Analysis in Subcellular Compartments of Arabidopsis Leaf Tissues
    [Abstract] The accurate determination of metabolite distribution in subcellular compartments is still challenging in plant science. Various methodologies, such as fluorescence resonance energy transfer-based technology, nuclear magnetic resonance spectroscopy and protoplast fractionation allow the study of metabolite compartmentation. However, large changes ...
    Simple Method to Determine Protein Redox State in Arabidopsis thaliana
    Authors:  Keisuke Yoshida and Toru Hisabori, date: 06/05/2019, view: 3134, Q&A: 0
    [Abstract] Thiol-based redox regulation is a posttranslational protein modification that plays a key role in many biological aspects. To understand its regulatory functions, we need a method to directly assess protein redox state in vivo. Here we present a simple procedure to determine protein redox state in a model plant Arabidopsis thaliana ...
    Quantification of the Humidity Effect on HR by Ion Leakage Assay
    Authors:  Musoki Mwimba and Xinnian Dong, date: 04/05/2019, view: 2607, Q&A: 0
    [Abstract] We describe a protocol to measure the contribution of humidity on cell death during the effector-triggered immunity (ETI), the plant immune response triggered by the recognition of pathogen effectors by plant resistance genes. This protocol quantifies tissue cell death by measuring ion leakage due to loss of membrane integrity during the ...
    Visualization of Plant Cell Wall Epitopes Using Immunogold Labeling for Electron Microscopy
    Author:  Mateusz Majda, date: 04/05/2019, view: 3411, 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 ...



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