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This volume provides updated technical approaches that have been developed to characterize monoterpene indole alkaloid metabolism in C. roseus from metabolite/gene product localization, alkaloid chemical synthesis, candidate gene prediction, transcription factor characterization up to functional genomic tools based on gene overexpression. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, Catharanthus roseus: Methods and Protocols aims to be a guidebook to all researchers working at characterizing alkaloid biosynthesis and more broadly specialized metabolisms
List of contents
RNA in situ hybridization on paraffin-sections to characterize the multicellular compartmentation of plant secondary metabolisms.- Imaging MS analysis in Catharanthus roseus.- Surface-Assisted Laser Desorption/Ionization Imaging Mass Spectrometry (SALDI-IMS)-Based Detection of Vinca Alkaloids Distribution in the Petal of Madagascar Periwinkle.- Sample preparation, data acquisition, and data analysis for 15N-labeled and non-labeled monoterpene indole alkaloids in Catharanthus roseus.- Studying iridoid transport in Catharanthus roseus by grafting.- Pictet-Spengler reaction for the chemical synthesis of strictosidine.- Implementation of a MS/MS spectral library for Monoterpene Indole Alkaloids.- Semi-Synthesis of Bis-Indole Alkaloid (-)-Melodinine K Enabled by a Combination of Biotransformation and Chemical Synthesis.- RNA-seq analysis towards monoterpene indole alkaloid biosynthetic pathway elucidation in Catharanthus roseus.- Predicting Monoterpene Indole Alkaloid related genes from expression data with artificial neural networks.- Discovery and characterization of oxidative enzymes in monoterpenoid indole alkaloid biosynthesis.- Ancestral sequence reconstruction for exploring alkaloid evolution.- Generating an EMS mutant population and rapid mutant screening by thin layer chromatography enables the studies of monoterpenoid indole alkaloids biosynthesis in Catharanthus roseus.- TARGETing transcriptional regulation in the medicinal plant Catharanthus roseus.- Identification and Characterization of Transcription Factors Regulating Terpenoid Indole Alkaloid Biosynthesis in Catharanthus roseus.- From Methylome to Integrative Analysis of Tissue-Specificity.- Tagging and capture of prenylated CaaX-proteins from plant cell cultures.- EASI transformation protocol: An Agrobacterium-mediated transient transformation protocol for Catharanthus roseus seedlings.- A rapid and efficient vacuum-based Agroinfiltration protocol for transient gene overexpression in leaves of Catharanthus roseus.- Transient gene expression in Catharanthus roseus flower petals using agroinfiltration.- De novo shoot bud induction from the Catharanthus roseus leaf explants and Agrobacterium tumefaciens mediated technique to raise transgenic plants.- Agrobacterium-mediated in planta transformation in Periwinkle.
