Plant Science

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    Protocols in Current Issue
    In vitro Assays to Evaluate Specificity and Affinity in Protein-phospholipid Interactions
    [Abstract]

    Protein–lipid interactions play important roles in many biological processes, including metabolism, signaling, and transport; however, computational and structural analyses often fail to predict such interactions, and determining which lipids participate in these interactions remains challenging. In vitro assays to assess the physical

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    Apoplastic Expression of CARD1-ecto Domain in Nicotiana benthamiana and Purification from the Apoplastic Fluids
    Authors:  Nobuaki Ishihama, Anuphon Laohavisit, Kaori Takizawa and Ken Shirasu, date: 04/20/2022, view: 767, Q&A: 0
    [Abstract]

    The protein expression and purification process is an essential initial step for biochemical analysis of a protein of interest. Traditionally, heterologous protein expression systems (such as E. coli, yeast, insect cells, and cell-free) are employed for plant protein expression, although a plant expression system is often desirable for plant

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    In vitro Auto- and Substrate-Ubiquitination Assays
    Authors:  Hye Lin Park, Han Yong Lee and Gyeong Mee Yoon, date: 04/05/2022, view: 446, Q&A: 0
    [Abstract]

    The precise regulation of the homeostasis of the cellular proteome is critical for the appropriate growth and development of plants. It also allows the plants to respond to various environmental stresses, by modulating their biochemical and physiological aspects in a timely manner. Ubiquitination of cellular proteins is one of the major

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    Activity-based Protein Profiling of Serine Hydrolase Superfamily Enzymes
    Authors:  Achintya Kumar Dolui, Mahadev Latha and Panneerselvam Vijayaraj, date: 03/20/2022, view: 510, Q&A: 0
    [Abstract]

    Activity-based protein profiling (ABPP) is a chemoproteomics platform to assess the functional state of enzymes in complex biological systems. Over the two decades, ABPP has emerged from a gel-based to gel-free platform, for in-depth proteome analysis with enhanced resolution, sensitivity for target detection, and discovery of small molecule

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    A Quick Method to Quantify Iron in Arabidopsis Seedlings
    Authors:  Chandan Kumar Gautam, Huei-Hsuan Tsai and Wolfgang Schmidt, date: 03/05/2022, view: 1119, Q&A: 0
    [Abstract]

    Iron (Fe) is an indispensable micronutrient for plant growth and development. Since both deficiency, as well as a surplus of Fe, can be detrimental to plant health, plants need to constantly tune uptake rates to maintain an optimum level of Fe. Quantification of Fe serves as an important parameter for analyzing the fitness of plants from different

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    Measuring Endogenous GA and IAA
    Authors:  Yunping Chen, Zhiwu Dan and Shaoqing Li, date: 02/20/2022, view: 1029, Q&A: 0
    [Abstract]

    Plant hormones regulate many physiological processes that largely influence growth, differentiation, and development. Contents of phytohormones were analyzed using a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) system. This protocol describes a detailed procedure to extract and quantify indole-3-acetic acid

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    An in vitro Coupled Assay for PEPC with Control of Bicarbonate Concentration
    Authors:  Nicholas R. Moody, Chatawal Phansopal and James D. Reid, date: 12/20/2021, view: 799, Q&A: 0
    [Abstract]

    Phosphoenolpyruvate carboxylase (PEPC) catalyzes a critical step in carbon metabolism in plants and bacteria, the irreversible reaction between bicarbonate and phosphoenolpyruvate to produce the C4 compound oxaloacetate. This enzyme is particularly important in the context of C4 photosynthesis, where it is the initial carbon-fixing enzyme. Many

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    Preference Test of Plutella xylostella Larvae upon DMNT Treatment
    Authors:  Chen Chen, Zhen Tao, Chuanhong Wang, Tengyue Wang, Hongyi Chen, Taoshan Jiang and Peijin Li, date: 11/05/2021, view: 886, Q&A: 0
    [Abstract]

    We describe a method to test the preference of insects in response to (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT). We use a device that includes a horizontal glass tube, two grooves (with activated carbon), air flow, rubber stoppers/tubes, transparent glass containers (optional), and a holder for the glass tube (optional). Equal amounts of activated

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    High Throughput Analyses of Ascorbate-turnover Enzyme Activities in Rice (Oryza sativa L.) Seedlings
    Authors:  Lin-Bo Wu, Yanru Feng, Frederike Zeibig, Muhammad Shahedul Alam and Michael Frei, date: 10/20/2021, view: 1198, Q&A: 0
    [Abstract]

    Ascorbate (Vitamin C) fulfills various functions in plant photosynthesis and abiotic stress tolerance. The four key enzymes involved in the ascorbate-turnover pathway are ascorbate peroxidase, ascorbate oxidase, monodehydroascorbate reductase, and dehydroascorbate reductase. Several reports have shown the pivotal roles of these enzymes in plant

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    Proteoliposomes for Studying Lipid-protein Interactions in Membranes in vitro
    Author:  Helmut Kirchhoff, date: 10/20/2021, view: 935, Q&A: 0
    [Abstract]

    Lipids in biomembranes can control the structure and, therefore, the functionality of membrane-embedded protein complexes. Unraveling how the lipid composition determines the mode of operation of membrane proteins provides mechanistic insights into their functionality. We applied a proteoliposome technique for studying how proteins function in

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