1. Genetic Screens to Target Embryo and Endosperm Pathways in Arabidopsis and Maize
C. Stewart Gillmor, A. Mark Settles, and Wolfgang Lukowitz
2. EMS Mutagenesis of Arabidopsis Seeds
C. Stewart Gillmorand Wolfgang Lukowitz
3. EMS Mutagenesis of Maize Pollen
A. Mark Settles
Part II: Microscopic Imaging and Phenotypic Analysis of Arabidopsis Embryos
4. Live-cell Imaging of Zygotic Intracellular Structures and Early Embryo Pattern Formation in Arabidopsis thaliana
Minako Ueda, Yusuke Kimata, and Daisuke Kurihara
5. Analyzing Sub-Cellular Reorganization during Early Arabidopsis Embryogenesis using Fluorescent Markers
Che-Yang Liao and Dolf Weijers
6. From Stained Plant Tissues to Quantitative Cell Segmentation Analysis with MorphoGraphX
Merijn Kerstens, Soeren Strauss, Richard Smith, and Viola Willemsen
Part III: Expression Analysis and Transcriptomics
7. Small RNA In Situ Hybridizations on Sections of Arabidopsis Embryos
Katalin Páldi, Magda Mosiolek, and Michael D. Nodine
8. Manual Isolation of Living Early Embryos from Tobacco Seeds
Peng Zhao, Xuemei Zhou, Ce Shi, and Meng-xiang Sun
9. Profiling Transcriptomes of Manually Dissected Arabidopsis Embryos
Ping Kao and Michael D. Nodine
10. Laser-assisted Microdissection of Plant Embryos for Transcriptional Profiling
Ana Marcela Florez-Rueda, Lucas Waser, and Ueli Grossniklaus
11. Cell-type-specific Transcriptomics in the Plant Embryo Using an Adapted INTACT Protocol
Joakim Palovaara and Dolf Weijers
12. Transcriptomic Profiling of the Arabidopsis Embryonic Epidermis Using FANS in Combination with RNAseq
Daniel Slane, Kenneth W. Berendzen, Janika Witthöft, and Gerd Jürgens
Part IV: Protein Interactions Studies
13. Visualizing Protein Associations in living Arabidopsis Embryo
Yuchen Long,Yvonne Stahl,Stefanie Weidtkamp-Peters, and Ikram Blilou
Part V: Embryogenesis in Other Model and Non-Model Species
14. Imaging of Embryo Sac and Early Seed Development in Maize after Feulgen Staining
Kamila Kalinowska, Junyi Chen, and Thomas Dresselhaus
15. Using Giant Scarlet Runner Bean (Phaseolus coccineus) Embryos to Dissect the Early Events in Plant Embryogenesis
Min Chen, Anhthu Q. Bui, and Robert B. Goldberg
16. Microscopical Detection of Cell Death Processes during Scots Pine Zygotic Embryogenesis
Jaana Vuosku and Suvi Sutela
Part VI: In Vitro Systems to Study Embryogenesis
17. Regulation of Somatic Embryo Development in Norway Spruce
Sara von Arnold, Tianqing Zhu, Emma Larsson, Daniel Uddenberg, and David Clapham
18. In vitro Production of Zygotes by Electro-Fusion of Rice Gametes
Md Hassanur Rahman, Erika Toda, and Takashi Okamoto
19. Isolated Microspore Culture in Brassica napus
Patricia Corral-Martínez, Carolina Camacho-Fernández, and Jose M. Seguí-Simarro
20. Anther Culture in Eggplant (Solanum melongena L.)
Antonio Calabuig-Serna, Rosa Porcel, Patricia Corral-Martínez, and Jose M. Seguí-Simarro
This volume details state-of-the-art methods for the study of plant embryogenesis in the model organism Arabidopsis thaliana, other models, and non-model species. Chapters guide readers through genetic screens, phenotypic analysis, live imaging, transcriptional profiling, methods on other model and non-model species beyond Arabidopsis thaliana, and introduction to systems that allow to culture or produce zygotic and somatic embryos in vitro. 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.
Authoritative and cutting-edge, Plant Embryogenesis: Methods and Protocols aims to ensure successful results in the further study of this vital field.
The chapter “Small RNA In Situ Hybridizations on Sections of Arabidopsis Embryos” is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.