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This volume provides a collection of protocols aimed toward the study of ethylene signaling in plants. Ethylene Signaling: Methods and Protocols is divided into three sections: ethylene biosynthesis, the signal transduction pathway, and the diverse ethylene responses of dicots and monocots. The chapters in section one discuss techniques for the measurement of activities related to the biosynthetic enzymes ACC synthase and ACC oxidase, the levels of ethylene synthesized by plants, and the treatment of plants with exogenous ethylene. Section two focuses on the analysis of the new membrane-associated proteins involved in the initial perception and transduction of the ethylene signal, such as ethylene receptors, CTR1, and EIN2. The third section covers assays applicable to dicots and monocots, including methods related to the roles of ethylene in germination, growth, abscission, abiotic stress, and defense. Section three also includes information on Arabidopsis mutants and the variety of chemical inhibitors that affect ethylene responses. 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.

Thorough and comprehensive, Ethylene Signaling: Methods and Protocols is a valuable resource for both experienced and beginner researchers with prior experience in the study of ethylene signaling and for those who are just entering this exciting research field.

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Preface...
Table of Contents...
Contributing Authors...

Part I Analysis of Ethylene Biosynthesis

1. Gas Chromatography-Based Ethylene Measurement of Arabidopsis Seedlings
Gyeong Mee Yoon and Yi-Chun Chen

2. Plant Ethylene Detection Using Laser Based Photo-Acoustic Spectroscopy
Bram Van de Poel and Dominique Van Der Straeten

3. Treatment of Plants with Gaseous Ethylene and Gaseous Inhibitors of Ethylene Action
Mark L. Tucker, Joonyup Kim, and Chi-Kuang Wen

4. Analysis of 1-Aminocyclopropane-1-Carboxylic Acid Uptake Using a Protoplast System
Won-Yong Song, Sumin Lee, and Moon-Soo Soh

5. Escherichia coli-Based Expression and In Vitro Activity Assay of 1-Aminocyclopropane-1-Carboxylate (ACC) Synthase and ACC Oxidase
Shigeru Satoh and Yusuke Kosugi

6. Assay Methods for ACS Activity and ACS Phosphorylation by MAP Kinases In Vitro and In Vivo
Xiaomin Han, Guojing Li, and Shuqun Zhang

Part II Analysis of the Ethylene Signaling Pathway

7. Analysis of Ethylene Receptors: Ethylene Binding Assays
Brad M. Binder and G. Eric Schaller

8. Analysis of Ethylene Receptors: Assay for Histidine Kinase Activity
G. Eric Schaller and Brad M. Binder

9. Analysis of Ethylene Receptor Interactions by Co-Immunoprecipitation Assays
Zhiyong Gao and G. Eric Schaller

10. Localization of the Ethylene-Receptor Signaling Complex to the Endoplasmic Reticulum: Analysis by Two-Phase Partitioning and Density Gradient Centrifugation
G. Eric Schaller
11. Kinase Assay for CONSTITUTIVE TRIPLE RESPONSE 1 (CTR1) in Arabidopsis thaliana
Han Yong Lee and Gyeong Mee Yoon

12. Circular Dichroism and Fluorescence Spectroscopy to Study Protein Structure and Protein-Protein Interactions in Ethylene Signaling
Mareike Kessenbrock and Georg Groth

Part III Analysis of Ethylene Responses

13. The Triple Response Assay and Its Use to Characterize Ethylene Mutants in Arabidopsis
Catharina Merchante and Anna N. Stepanova

14. Time-Lapse Imaging to Examine the Growth Kinetics of Arabidopsis Seedlings in Response to Ethylene
Brad M. Binder

15. Inhibitors of Ethylene Biosynthesis and Signaling
G. Eric Schaller and Brad M. Binder

16. Analysis of Growth and Molecular Responses to Ethylene in Etiolated Rice Seedlings
Biao Ma and Jin-Song Zhang

17. Love Me Not Meter: A Sensor Device for Detecting Petal Detachment Forces in Arabidopsis thaliana
Andrew Maule, Graham Henning, and Sara Patterson

18. Effects of Ethylene on Seed Germination of Halophyte Plants under Salt Stress
Weiqiang Li and Lam-Son Phan Tran

19. Assessing Attraction of Nematodes to Host Roots using Pluronic Gel Medium
Valerie M. Williamson and Rasa Cepulyt-Rakauskien

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