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This volume provides a comprehensive compilation of techniques and protocols used in plant and food carotenoid research. Chapters guide readers through seven major areas on core enzyme activities, analysis of carotenoid profiles, new imaging tools, synthesis and degradation dynamics, biotechnology, nutrition, and health. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Plant and Food Carotenoids: Methods and Protocols aims to be helpful to researchers of other disciplines that are impacted by carotenoids, including photosynthesis, biotechnology, food science, and nutrition.
Sommario
Pathways for Carotenoid Biosynthesis, Degradation, and Storage.- A Simple in vitro Assay to Measure the Activity of Geranylgeranyl Diphosphate Synthase and Other Short-chain Prenyltransferases.- Expression, Purification, and Enzyme Activity Assay of Phytoene Synthase in vitro.- Improved Expression and Purification of the Carotenoid Biosynthetic Enzyme Z-ISO.- Determination of In Vitro and In Vivo Activities of Plant Carotenoid Cleavage Oxygenases.- Expression and Characterization of Mammalian Carotenoid Cleavage Dioxygenases.- Transportomics for the Characterization of Plant Apocarotenoid Transmembrane Transporters.- Applications of Visible Spectroscopy and Colour Measurements in the Assessments of Carotenoid Levels in Foods.- A Routine Method for the Extraction and HPLC-DAD Profiling of Major Plant and Food Carotenoids.- Acquisition of Mass Spectrometry Data of Carotenoids: A Focus on Big Data Management.- cis/trans Carotenoid Extraction, Purification, Detection, Quantification, and Profiling in Plant Tissues.- Determination of Plant Volatile Apocarotenoids.- A Method for Extraction and LC-MS Based Identification of Carotenoid-Derived Dialdehydes in Plants.- Fractionation of Tomato Fruit Chromoplasts.- Quantification of Strigolactones.- Carotenoid and Apocarotenoid Analysis by SFE-SFC-QqQ/MS.- Elucidating Carotenoid Biosynthetic Enzyme Localization and Interactions Using Fluorescent Microscopy.- Visualization of Carotenoid-storage Structures in Fruits by Transmission Electron Microscopy.- Light Microscopy and Raman Imaging of Carotenoids in Plant Cells in situ and in Released Carotene Crystals.- Carotenoid Isotopologue Profiling in 13C-labeled Leaf Extracts by LC-MS and LC-FTICR-MS.- Quantification of Carotenoid Pathway Flux in Green and Non-green Systems.- Histochemical and Molecular Quantification of Arbuscular Mycorrhiza Symbiosis.- Use of Potyvirus Vectors to Produce Carotenoids in Plants.- Agrobacterium tumefaciens Mediated Stable Transformation of Daucus carota.- CRISPR-Cas-Mediated Gene Knockout in Tomato.- Modulation of Activity of a Carotenoid Pathway Through the use of the TET-on Regulatory System: Application in the Fungus Fusarium fujikuroi.- Intervention Studies in Humans.- In vitro Digestion for Control and Monitoring of Food Effects In Relation To Micellarization Index of Carotenoids.- Evaluation of Carotenoids Protection against Oxidative Stress in the Animal Model Caenorhabditis elegans.- Mouse Models to Study Antiobesogenic Effects of Carotenoids.
