Plant Proteomics

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The aim of this work is to present up-to-date methods and protocols used by leading scientists in plant proteomics. Over the past twenty-five years, this field has grown exponentially, with laboratories worldwide focusing on plant proteomics. Two-dimensional gel electrophoresis remains a fundamental technique, enhanced by improvements such as IPG in the first dimension and new detection methods using fluorochromes. Significant advancements have been made in protein extraction, particularly from plant tissues that contain phenols, proteases, and other secondary metabolites that complicate protein analysis. Standard procedures have been optimized for specific tissues and cellular compartments. These methods leverage improvements in protein solubilization from membranes. Mass spectrometry has revolutionized the high-throughput identification of proteins separated by 2D gels, as well as from blue native 1D gels, while Edman sequencing continues to have its applications. Coupled with techniques like 2DLC or LC of intact proteins, mass spectrometry enables the identification of polypeptides from complexes, showcasing the continuous evolution and sophistication of methodologies in plant proteomics.