Pollen and Pollen Tube Biology, 1st ed. 2020
Methods and Protocols
Methods in Molecular Biology Series, Vol. 2160

This volume explores a collection of experimental techniques used to investigate different aspects of pollen development and function, including its role in reaching the ovule and delivering the two sperm cells. The techniques discussed range from basic methodology to cultivate pollen in vitro to the sophisticated experiments involving micromanipulation, Lab-on-Chip technology, or high-end imaging. The chapters in this book cover topics such as pollen grain counting using a cell counter; restricted pollination for tracing individual pollen tubes in a pistil; obtaining mutant pollen for phenotypic analysis and pollen tube dual-staining; analyzing intracellular gradients in pollen tubes; and measuring exocytosis rate in Arabidopsis pollen tube using corrected fluorescence recovery after photoconversion (cFRAPc) technique. 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.

Cutting-edge and thorough, Pollen and Pollen Tube Biology: Methods and Protocols is a valuable resource for novice and expert scientists interested in learning more about the field of plant reproduction. 

Preface…
Table of Contents…
Contributing Authors…

1. Pollen Grain Counting Using a Cell Counter
Hiroyuki Kakui, Misako Yamazaki, Naoto-Benjamin Hamaya, and Kentaro K. Shimizu

2. A Toolkit for teasing Apart the Early Stages of Pollen-Stigma Interactions in Arabidopsis thaliana
Hyun Kyung Lee, Stuart Macgregor, and Daphne R. Goring

3. Decapitation Crosses to Test Pollen Fertility Mutations for Defects in Stigma-Style Penetration
Chrystle Weigand and Jeffrey Harper

4. Isolation of the Pistil-Stimulated Pollen Tube Secretome
Said Hafidh and David Honys

5. Restricted Pollination for Tracing Individual Pollen Tubes in a Pistil
Taro Takahashi, Toshiyuki Mori, and Tomoko Igawa

6. Semi-In Vivo Assay for Pollen Tube Attraction
Sheng Zhong, Zhijuan Wang, and Li-Jia Qu

7. Isolation and Cloning of Suppressor Mutants with Improved Pollen Fertility
Steven Beuder and Cora A. MacAlister

8. Workflow to Characterize Mutants with Reproductive Defects
Jennifer A. Noble and Ravishankar Palanivelu

9. Internally-Controlled Methods to Quantify Pollen Tube Growth and Penetration Defects in Arabidopsis thaliana
Devin K. Smith and Ian S. Wallace

10. Arabidopsis thaliana Pollen Tube Culture for Multi-Omics Studies
M. Costa, J. Silva, and S. Coimbra

11. Large Scale Analysis of Pollen Viability and Oxidative Level using H2DCFDA-Staining Coupled with Flow Cytometry
Nicholas Rutley and Gad Miller

12. Obtaining Mutant Pollen for Phenotypic Analysis and Pollen Tube Dual-Staining
Sheng Zhong, Zhijuan Wang, and Li-Jia Qu

13. Galvanotropic Chamber for Controlled Reorientation of Pollen Tube Growth and Simultaneous Confocal Imaging of Intracellular Dynamics
Firas Bou Daher and Anja Geitmann

14. Analyzing Intracellular Gradients in Pollen Tubes
Daniel S. C. Damineli, Maria Teresa Portes, and José A. Feijó

15. Silicone Chambers for Pollen Tube Imaging in Microstructures In Vitro Environments
Hana Bertrand-Rakusová, Youssef Chebli, and Anja Geitmann

16. Analyzing the Impact of Protein Overexpression on Ca2+ Dynamics and Development in Tobacco Pollen Tubes
Chunxia Zhang, Leonie Steinhorst, and Jörg Kudla

17. Imaging and Analysis of the Content of Callose, Pectin, and Cellulose in the Cell Wall of Arabidopsis Pollen Tubes Grown In Vitro
Ana R. Sede, Diego L. Wengier, Cecilia Borassi, José M. Estevez, and Jorge P. Muschietti

18. Characterization of Growth Behavior and the Resulting Forces Applied by Pollen Tubes in a 3D Matrix
Ronny Reimann and Delf Kah

19. Flow Chamber Assay to Image the Response of FRET-Based Nanosensors in Pollen Tubes to Changes in Medium Composition
Theresa Maria Reimann

20. Quantification of Mechanical Forces and Physiological Processes Involved in Pollen Tube Growth Using Microfluidics and Microrobotics
Jan Burri, Gautam Munglani, Bradley J. Nelson, Ueli Grossniklaus, and Hannes Vogler

21. Measuring Exocytosis Rate in Arabidopsis Pollen Tube Using Corrected Fluorescence Recovery after Photoconversion (cFRAPc) Technique
Jingzhe Guo and Zhenbiao Yang

22. Testing Pollen Tube Proteins for In Vivo Binding to Phosphatidic Acid by n0Butanol Treatment and Confocal Microscopy
Carolin Fritz and Benedikt Kost

Subject Index List…

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts