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Isolation and Reproductive Structures Induction of Fungal Pathogens Associated with Xylem and Wood Necrosis in Grapevine
Grapevine (Vitis vinifera L.) trunk diseases (GTDs) are considered a disease complex including five diseases: esca, Petri disease, black-foot...
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Taxonomic Classification of Geminiviruses Based on Pairwise Sequence Comparisons
Geminiviridae is the largest and one of the most diverse families of plant viruses, comprising 14 genera demarcated based on host range, type of...
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Virus-Induced Heritable Gene Editing in Plants
Gene editing using clustered, regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) nuclease is an excellent tool for...
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Monitoring the Intracellular Trafficking of Virus-Induced Structures and Intercellular Spread of Viral Infection in Plants Using Endomembrane Trafficking Pathway-Specific Chemical Inhibitor and Organelle-Selective Fluorescence Dye
Infection by positive-strand RNA viruses induces extensive remodeling of the host endomembrane system in favor of viral replication and movement. The...
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Construction of Geminivirus Infectious Clones for Agroinoculation into Plants
The production of geminiviral infectious clones provides a standardized inoculum for use in several host–virus studies. Geminiviruses present either...
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Co-immunoprecipitation Assays to Detect Protein–Protein Interactions
Proteins usually do not function as monomers but rather perform their functions by interacting with themselves or other proteins....
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Quantification of Xylem-Specific Thermospermine-Dependent Translation of SACL Transcripts with Dual Luciferase Reporter System
Thermospermine (Tspm) is a polyamine found to play a crucial role in xylem development in Arabidopsis thaliana. Tspm promotes the translation of the...
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Measurement of Phytochrome B Thermal Reversion Rates In Vivo
Thermal reversion of phytochromes is the light-independent but strongly temperature-dependent relaxation of the light-activated Pfr form of...
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Characterization of Thermoresponsive Photobody Dynamics
Photobodies (PBs) are subnuclear membraneless organelles that self-assemble via the condensation of the plant photoreceptor and thermosensor...
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GST Pull-Down Assay to Study PIF4 Binding In Vitro
The phytochrome-interacting factor 4 (PIF4) is a well-known transcription factor that plays a pivotal role in plant thermomorphogenesis, coordinating...
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RuBisCO Depletion and Subcellular Fractionation for Enhanced Florigen Detection in Arabidopsis
In plants, complex signaling networks monitor and respond to environmental cues to determine the optimal time for the transition from the vegetative...
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Measurement of Photorespiratory Oxygen Exchange by Membrane Inlet Mass Spectrometry
Photosynthesis and metabolism in plants involve oxygen as both a product and substrate. Oxygen is taken up during photorespiration and respiration...
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Combining Gas Exchange and Rapid Quenching of Leaf Tissue for Mass Spectrometry Analysis Directly in Gas Exchange Cuvette
Isotopically nonstationary metabolic flux analysis (INST-MFA) is a powerful technique for studying plant central metabolism, which involves...
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Analysis of Photorespiratory Intermediates Under Transient Conditions by Mass Spectrometry
Photorespiration is an essential process of phototropic organisms caused by the limited ability of rubisco to distinguish between CO2 and O2. To...
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Spectrophotometric Assays for Measuring Hydroxypyruvate Reductase Activity
Hydroxypyruvate reductase (HPR; EC 1.1.1.81) activity is integral to the photorespiratory pathway. Within photorespiration, HPR catalyzes the...
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Determination of Phosphoglycolate Phosphatase Activity via a Coupled Reaction Using Recombinant Glycolate Oxidase
Phosphoglycolate phosphatase (PGLP) dephosphorylates 2-phosphoglycolate to glycolate that can be further metabolized to glyoxylate by glycolate...
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Purification of Single-Stranded RNA Bacteriophages and Host Receptors for Structural Determination Using Cryo-Electron Microscopy
Single-stranded RNA bacteriophages (ssRNA phages) are small viruses with a compact genome (~3–4 kb) that infect gram-negative bacteria via retractile...
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Mitochondrial Dihydrolipoamide Dehydrogenase (mtLPD1): Expression, Purification, Activity, and Redox Regulation
Mitochondrial dihydrolipoamide dehydrogenase (mtLPD1) is a central enzyme in primary carbon metabolism, since its function is required to drive four...
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Purification of Recombinant N-terminal Histidine-Tagged Arabidopsis thaliana Phosphoglycolate Phosphatase 1, Glycolate Oxidase 1 and 2, and Hydroxypyruvate Reductase 1
To measure the kinetic properties of photorespiratory enzymes, it is necessary to work with purified proteins. Protocols to purify photorespiratory...
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Measurement of O2 Uptake and Evolution in Leaves In Vivo Using Stable Isotopes and Membrane Inlet Mass Spectrometry
Oxygen is both product and substrate of photosynthesis and metabolism in plants, by oxygen evolution through water splitting and uptake by...