Plant metabolomics methods and protocols
This book is part of the 'Methods in Molecular Biology' series (no. 860), which is popular for including chapters with introductory notes on the topic, lists of necessary materials and reagents, step-by-step detailed protocols, and key notes on troubleshooting and avoiding common errors. This volume describes several techniques, methods and experimental considerations that are commonly used in the field of plant metabolomics. The field of metabolomics and plant metabolomics has developed significantly during the last 15 years and this book includes contributions by leading authors in the area who played key roles in the development of this field. An introductory chapter describes the field of metabolomics and its growth in the past few years, defines some common terms, and presents the workflow commonly used in metabolomics experiments, from the experimental design to data analysis.The book is divided into three parts that target important topics in any given metabolomics experiment: material preparation, chemical analysis approaches and data analysis. The first chapter of the first section of the book (Chapter 2) includes guidelines on designing metabolomics experiments, not only with regard to the statistical design but also including considerations regarding environmental control, sampling strategy and size of experiment, and potential pitfalls associated with each of these. Chapter 3 presents an interesting protocol not only for the extraction of the metabolome of a plant but also microbial material, which could be applied in the field of plant-microbe interactions. The following chapter expands further on precautions necessary when sampling, transporting and storing biological material, which particularly apply in the field of plant metabolomics where it is common to sample material away from the laboratory. The first part of the book concludes with a chapter dedicated to a protocol for growing arabidopsis plants for metabolomics experiments, highlighting potential issues related to sample storage, sampling time-point strategies and considerations regarding the pooling of biological samples. The second part of the book is dedicated to chemical analysis approaches and contains a series of chapters that describe commonly used methods for the major analytical techniques in plant metabolomics, such as LC-MS and GC-MS. Protocols for GC-MS are presented in Chapters 6 and 7, which describe methods for analysing headspace volatiles and primary metabolites, respectively. This is followed by two chapters with protocols for liquid-chromatography-based mass-spectrometry, the first based on a HPLC system and the second describing a method based on UPLC. Chapters 10 and 11 describe the use of mass spectrometers (Orbitrap, FT-ICR-MS) able to generate an accurate mass fingerprint that can subsequently be used to identify plant metabolites. The following chapter describes a protocol for the extraction of Brassica green tissue and subsequent analysis using a combination of NMR and FI-ESI-MS.
| Main Authors: | , |
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| Format: | Texto biblioteca |
| Language: | eng |
| Published: |
New York, New York, United States Humana Press
c201
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| Subjects: | Metabolitos vegetales, Metabolismo vegetal, Análisis químico, Análisis de datos, |
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