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This volume describes up-to-date techniques for improved production of secondary metabolites of economic interest using field and laboratory methods. Biotechnology of Plant Secondary Metabolism: Methods and Protocols explores different secondary metabolite classes, whole-plant and cell/organ culture systems, and environmental and genetic transformation-based modulation of biochemical pathways. Special focus is given to cell and tissue specific metabolism, metabolite transport, microRNA-based technology, heterologous systems expression of enzymes and pathways leading to products of interest, as well as applications using both model and non-model plant species. 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.
Practical and cutting-edge, Biotechnology of Plant Secondary Metabolism: Methods and Protocols is a great resource for scientists of interdisciplinary fields--plant science, plant physiology, pharmacy, molecular biology, biochemistry, bioengineering, and forestry--in reaching their goals of producing plant biochemicals in a sustainable and efficient manner, while minimizing impacts to the environment and providing the required quantities of these commodities to industry.
Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras
Inhalt
Elicitation Approaches for Withanolide Production in Hairy Root Culture of Withania somnifera (L.) Dunal.- Stimulant Paste Preparation and Bark Streak Tapping Technique for Pine Oleoresin Extraction.- A Modified Protocol for High Quality RNA Extraction from Oleoresin-Producing Adult Pines.- Collection of Apoplastic Fluids from Arabidopsis thaliana Leaves.- From Plant Extract to a cDNA-Encoding a Glucosyltransferase Candidate : Proteomics and Transcriptomics as Tools to Help Elucidate Saponin Biosynthesis in Centella asiatica.- Production of Recombinant Caffeine Synthase from Guarana ( Paullinia cupana var. sorbillis ) in Escherichia coli.- Heterologous Expression and Characterization of Mimosinase from Leucaena leucocephala.- Production of Aromatic Plant Terpenoids in Recombinant Baker´s Yeast.- Purification of a Recombinant Polyhistidine-Tagged Glycosyltransferase Using Immobilized Metal-Affinity Chromatography (IMAC).- A Western Blot Protocol for Detection of Proteins Heterologously Expressed in Xenopus laevis Oocytes.- Laser Capture Microdissection: Avoiding Bias in Analysis by Selecting Just What Matters.- Analytical and Fluorimetric Methods for the Characterization of the Transmembrane Transport of Specialized Metabolites in Plants.- Protoplast Transformation as a Plant-Transferable Transient Expression System.- Design, Construction, and Validation of Artificial MicroRNA Vectors Using Agrobacterium -Mediated Transient Expression System.- Knockdown of Polyphenol Oxidase Gene Expression in Potato ( Solanum tuberosum L.) with Artificial MicroRNAs. <p