Plant Peptide Hormones and Growth Factors, 1st ed. 2024 Methods in Molecular Biology Series, Vol. 2731

This book describes state-of-the-art approaches for identification and functional characterization of plant peptide hormones and growth factors. Chapters guide readers through protocols for computational identification of novel signaling peptides and phytocytokines, analysis of post-translational maturation events, isolation of mature peptides from plant tissues, peptide bioassays, in vivo and in vitro analysis of peptide activity, peptide receptor identification and interaction analysis, and quantitative assessment of peptide ligand binding to cognate receptors. 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 key tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Plant Peptide Hormones and Growth Factors aims to be a useful and practical guide to new researchers and experts looking to expand their technical know-how.

Part I: Peptide Prediction, Formation and Isolation

1.   Bioinformatics Methods for Prediction of Gene Families Encoding Extracellular Peptides

Loup Tran Van Canh and Sébastien Aubourg

2. Identification of Bioactive Phytocytokines Using Transcriptomic Data and Plant Bioassays

Jack Rhodes and Cyril Zipfel

3. Purification of Phytaspases Using a Biotinylated Peptide Inhibitor

Raisa A. Galiullina, Ilya A. Dyugay, Andrey B. Vartapetian and Nina V. Chichkova

4. Characterization of Phytaspase Proteolytic Activity Using Fluorogenic Peptide Substrates

Raisa A. Galiullina, Nina V. Chichkova, Grigoriy G. Safronov and Andrey B. Vartapetian

5. Characterization of prolyl-4 hydroxylase substrate specificity using Pichia pastoris as an effective eukaryotic expression system

Gerith Elsäßer, Tim Seidl, Jens Pfannstiel, Andreas Schaller and Nils Stührwohldt

6. Extraction of Apoplastic Peptides for the Structural Elucidation of Mature Peptide Hormones in Arabidopsis

Mari Ogawa-Ohnishi and Yoshikatsu Matsubayashi

Part II: Peptide Activity and Signaling

Nanna Weise Havshøi and Anja Thoe Fuglsang

8. A Seedling Growth Inhibition Assay to Measure Phytocytokine Activity

Henriette Leicher and Martin Stegmann

9. A Trojan Horse Aproach Using Ustilago maydis to Study Apoplastic Maize (Zea mays) Peptides in situ

Leon Kutzner and Karina van der Linde

10. Feeding Assay to Study the Effect of Phytocytokines on Direct and Indirect Defense in Maize

Lei Wang and Matthias Erb

11. A Quick Method to Analyze Peptide-Regulated Anthocyanin Biosynthesis

Eric Bühler, Andreas Schaller and Nils Stührwohldt

12. Quantitative Measurement of Pattern-Triggered ROS Burst as an Early Immune Response in Tomato

Rong Li, Andreas Schaller and Annick Stintzi

13. Automated Real-Time Monitoring of Extracellular pH to Assess Early Plant Defense Signaling

Xu Wang, Rong Li, Annick Stintzi and Andreas Schaller

14. Peptide-Mediated Cyclic Nucleotide Signaling in Plants: Identification and Characterization of Interactor Proteins with Nucleotide Cyclase Activity

Ilona Turek and Chris Gehring

15. Detection of Ligand-Induced Receptor Kinase and Signaling Component Phosphorylation With Mn²⁺-Phos-Tag SDS-PAGE

Zunyong Liu, Shuguo Hou, and Ping He

Part II: Peptide - Receptor Interaction

16. In-vivo Cross Linking of Biotinylated Peptide Ligands to Cell Surface Receptors

Ronja Burggraf and Markus Albert

17. Evaluation of Direct Ligand-Receptor Interactions by Photoaffinity Labeling

Hidefumi Shinohara and Yoshikatsu Matsubayashi

18. Rapid Identification of Peptide-Receptor-Coreceptor Complexes in Protoplasts

Xiaoyang Wang and Xiangzong Meng

19. Acridinium-Based Chemiluminescent Receptor-Ligand Binding Assay for Protein/Peptide Hormones

André Guilherme Daubermann, Keini Dressano, Paulo Henrique de Oliveira Ceciliato and Daniel S. Moura

20. LuBiA (Luciferase-based Binding Assay): Glowing Peptides as Sensitive Probes to Study Ligand-Receptor Interactions

Louis-Philippe Maier, Georg Felix, Judith Fliegmann

21. Microscale Thermophoresis (MST) to Study Rapid Alkalinization Factor (RALF) - Receptor Interactions

Martine Gonneau, Sébastjen Schoenaers, Caroline Broyart, Kris Vissenberg, Julia Santiago and Herman Höfte

22. Isothermal Titration Calorimetry to Study Plant Peptide Ligand-Receptor Interactions

Judith Lanooij and Elwira Smakowska-Luzan

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts