Plant Germline Development, 1st ed. 2017 Methods and Protocols Methods in Molecular Biology Series, Vol. 1669

Part I: Plant Germline Development: From Investigation to Application

1. An Introduction to Male Germline Development

            Hannes Vogler, Ueli Grossniklaus, and Afif Hedhly

2. Apomixis: Engineering the Ability to Harness Hybrid Vigor in Crop Plants

            Joann A. Conner and Peggy Ozias-Akins

Part II: Clearing, Imaging Techniques, and Immunolocalization to Study Cell Lineage, Cell Type Specificity, Cytoskeleton, Cell Cycle, Signaling, and Hormones

3. 3D Imaging of Whole-Mount Ovules at Cellular Resolution to Study Female Germline Development in Rice

            Ethel Mendocilla Sato, Wenjing She, and Célia Baroux

4. Live-Cell Imaging of F-Actin Dynamics During Fertilization in Arabidopsis thaliana

            Daichi Susaki, Daisuke Maruyama, Ramesh Yelagandula, Frederic Berger, and Tomokazu Kawashima

5. Development and Observation of Mature Megagametophyte Cell-Specific Fluorescent Markers

            Mark A. Chamberlin and Shai J. Lawit

6. Analysis of Fluorescent Reporter Activity in the Male Germline During Pollen Development by Confocal Microscopy

            David Twell and Lynette Brownfield

7. In Vivo Ploidy Determination of Arabidopsis thaliana Male and Female Gametophytes

            Burcu Nur Keçeli, Nico De Storme, and Danny Geelen

8. Staining and Clearing of Arabidopsis Reproductive Tissue for Imaging of Fluorescent Proteins

            Daniel Slane, Patrick Bürgel, and Martin Bayer

9. Live Cell Imaging of Auxin and Cytokinin Signaling in Maize Female Gametophytes

            Antony M. Chettoor and Matthew M.S. Evans

10. Imaging Ca²⁺ Dynamics in Wild-Type and NADPH Oxidase-Deficient Mutant Pollen Tubes with Yellow Cameleon and Confocal Laser Scanning Microscopy

            Christina Maria Franck, Jens Westermann, and Aurélien Boisson-Dernier

11. Immunolocalization of AGPs and Pectins in Quercus suber Gametophytic Structures

            Mário Luís da Costa, Ana Lúcia Lopes, Maria Isabel Amorim, and Sílvia Coimbra

Part III: Chromosome Spreading, Whole Mount In Situ, and Transmission Electron Microscopy (TEM)

            Laksana Kantama, Erik Wijnker, and Hans de Jong

13. Whole Mount RNA-FISH on Ovules and Developing Seeds

            Andrea Bleckmann and Thomas Dresselhaus

14. Analysis of Peroxisome Biogenesis in Pollen by Confocal Microscopy and Transmission Electron Microscopy

            Peng-Fei Jia, Hong-Ju Li, and Wei-Cai Yang

15. Transmission Electron Microscopy (TEM) to Study Histology of Pollen and Pollen Tubes

            Peng-Fei Jia, Hong-Ju Li, and Wei-Cai Yang

Part IV: Isolation of Specific Cells of the Male and Female Germlines

16.Isolation of Arabidopsis Pollen, Sperm Cells, and Vegetative Nuclei by Fluorescence-Activated Cell Sorting (FACS)

            Mário R. Santos, Cláudia Bispo, and Jörg D. Becker

17. Isolation of Rice Sperm Cells for Transcriptional Profiling

            Scott D. Russell, Daniel S. Jones, Sarah Anderson, Xinkun Wang, Venkatesan Sundaresan, and Xiaoping Gou

18. Manual Isolation of Living Cells from the Arabidopsis thaliana Female Gametophyte by Micromanipulation

            Maria Englhart, Lucija Šoljić, and Stefanie Sprunck

Part V: Omics Technologies and Bioinformatic Analyses

19. Isolation and Detection of Small RNAs from Pollen

            German Martinez

20. Analysis of Proteins Enriched in Rice Gamete

            Takashi Okamoto

21. Phosphoprotein Enrichment from Tobacco Mature Pollen Crude Protein Extract

            Jan Fíla and David Honys

22. Identification of Cis-Regulatory Modules that Function in the Male Germline of Flowering Plants

            Benjamin Peters, Jack Aidley, Murray Cadzow, David Twell, and Lynette Brownfield

23. RNA-Seq Data Analysis: From Raw Data Quality Control to Differential Expression Analysis

24. RNA-Seq Data Analysis Protocol: Combining In-House and Publicly Available Data

            Marc W. Schmid

Part VI: Technologies with Outreach for Plant Manipulation, Studies of Pathways, and Applications in Crop Species

25. Impedance Flow Cytometry as a Tool to Analyze Microspore and Pollen Quality

            Iris Heidmann and Marco Di Berardino

26. Agrobacterium-Mediated Sorghum Transformation

            Emily Wu and Zuo-Yu Zhao

27. Use of the Cas9 Orthologues from Streptococcus thermophilus and Staphylococcus aureus for Non-Homologous End-Joining Mediated Site-Specific Mutagenesis in Arabidopsis thaliana

            Jeannette Steinert, Carla Schmidt, and Holger Puchta

28. Transgenic Reproductive Cell Ablation

29. In Vivo Reporters for Protein Half-Life

            Pavel Reichman and Nico Dissmeyer

30. Conditional Modulation of Biological Processes by Low-Temperature Degrons

            Nico Dissmeyer

Includes cutting-edge methods and protocols for the study of plant germline development

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts