What Is New in (Plant) Proteomics Methods and Protocols: The 2015–2019 Quinquennium

The third edition of "Plant Proteomics Methods and Protocols," with the title "Advances in Proteomics Techniques, Data Validation, and Integration with Other Classic and -Omics Approaches in the Systems Biology Direction," was conceived as being based on the success of the previous editions, and the continuous advances and improvements in proteomic techniques, equipment, and bioinformatics tools, and their uses in basic and translational plant biology research that has occurred in the past 5 years (in round figures, of around 22,000 publications referenced in WoS, 2000 were devoted to plants).The monograph contains 29 chapters with detailed proteomics protocols commonly employed in plant biology research. They present recent advances at all workflow stages, starting from the laboratory (tissue and cell fractionation, protein extraction, depletion, purification, separation, MS analysis, quantification) and ending on the computer (algorithms for protein identification and quantification, bioinformatics tools for data analysis, databases and repositories). At the end of each chapter there are enough explanatory notes and comments to make the protocols easily applicable to other biological systems and/or studies, discussing limitations, artifacts, or pitfalls. For that reason, as with the previous editions, it would be especially useful for beginners or novices.Out of the 29 chapters, six are devoted to descriptive proteomics, with a special emphasis on subcellular protein profiling (Chapters 5 - 10 ), six to PTMs (Chapters 11 , and 14 - 18 ), three to protein interactions (Chapters 19 - 21 ), and two to specific proteins, peroxidases (Chapter 24 ) and proteases and protease inhibitors (Chapter 26 ). The book reflects the new trajectory in MS-based protein identification and quantification, moving from the classic gel-based approaches to the most recent labeling (Chapters 10 , 11 , 29 ), shotgun (Chapters 5 , 7 , 12 , 15 ), parallel reaction monitoring (Chapter 16 ), and targeted data acquisition (Chapter 13 ). MS imaging (Chapter 25 ), the only in vivo MS-based proteomics strategy, is far from being fully optimized and exploited in plant biology research. A confident protein identification and quantitation, especially in orphan species, of low-abundance proteins, is still a challenging task (Chapters 4 , 28 ).What is really new is the use of different techniques for proteomics data validation and their integration into other classic and -omics approaches in the systems biology direction. Chapter 2 reports on multiple extractions in a single experiment of the different biomolecules, nucleic acids, proteins, and metabolites. Chapter 27 describes how metabolic pathways can be reconstructed from multiple -omics data, and Chapter 3 network building. Finally, Chapters 22 and 23 deal with, respectively, the search for allele-specific proteins and proteogenomics.Around 200 groups were, almost 1 year ago, invited to take part in this edition. Unfortunately, only 10% of them kindly accepted. My gratitude to those who accepted our invitation but also to those who did not, as all of them have contributed to the plant proteomics field. I will enlist, in this introductory chapter, following my own judgment, some of the relevant papers published in the past 5 years, those that have shown us how to enhance and exploit the potential of proteomics in plant biology research, without aiming at giving a too exhaustive list.