| Description |
Flow cytometry is widely used to estimate ploidy levels and genome sizes in plants and animals. It is also a common tool for determining the reproductive mode in flowering plants through flow cytometric seed screening (FCSS), which can differentiate sexual and apomictic pathways. However, FCSS has limitations, as it is not universally applicable to all apomictic taxa and cannot resolve certain reproductive pathways. These challenges can be addressed by germinating seeds and genotyping seedlings and their parents. However, this approach can be time- and space-intensive and carry other drawbacks (e.g., the ploidy level of the endosperm remains undetected). Here, we present a streamlined method that combines flow cytometry with genotyping of the same seeds. Applying both methods to a single seed enables comprehensive insights into reproductive modes, detecting various forms of apomixis and automixis, assessment of the level of selfing, regardless of the mode of reproduction mode, and identifying pollen donors in the endosperm of apomictic seeds. Each protocol—FCSS and seed genotyping—can also be used independently to determine basic reproductive modes. This volume explores the latest research techniques (and reviews) used to study apomixis model species. The chapters in this book are organized into six parts: Part One introduces gametophytic apomixis, and Part Two focuses on the detection and characterization of apomixis. Part Three talks about transcriptomic profiling of apomictic and sexual species, and Part Four covers genetic analysis of apomixis. Part Five is dedicated to research on model apomictic systems such as dandelion (Taraxacum), Pilosella, Boechera, and Cenchrus (Pennisetum), and the final Part Six looks at protocols for engineering either individual components of apomixis, such as apomeiosis or a complete synthetic apomixis system. 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 comprehensive, Apomixis: Methods and Protocols is a valuable reference for studying apomixis, and serves as an inspiration and foundation for designing new approaches and achieving significant breakthroughs in the field. This volume explores the latest research techniques (and reviews) used to study apomixis model species. The chapters in this book are organized into six parts: Part One introduces gametophytic apomixis, and Part Two focuses on the detection and characterization of apomixis. Part Three talks about transcriptomic profiling of apomictic and sexual species, and Part Four covers genetic analysis of apomixis. Part Five is dedicated to research on model apomictic systems such as dandelion (Taraxacum), Pilosella, Boechera, and Cenchrus (Pennisetum), and the final Part Six looks at protocols for engineering either individual components of apomixis, such as apomeiosis or a complete synthetic apomixis system. 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 comprehensive, Apomixis: Methods and Protocols is a valuable reference for studying apomixis, and serves as an inspiration and foundation for designing new approaches and achieving significant breakthroughs in the field.
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