Flow Cytometry With Plant Cells

Targeted at beginners as well as experienced users, this handy reference explains the benefits and uses of flow cytometery in the study of plants and their genomes. Following a brief introduction that highlights general considerations when analyzing plant cells by flow cytometric methods, the book goes on to discuss examples of application in plant genetics, genomic analysis, cell cycle analysis, marine organism analysis and breeding studies. With its list of general reading and a glossary of terms, this first reference on FCM in plants fills a real gap by providing first-hand practical hints for the growing community of plant geneticists.

The production of doubled haploids has become a necessary tool in advanced plant breeding institutes and commercial companies for breeding many crop species. However, the development of new, more efficient and cheaper large scale production protocols has meant that doubled haploids are also recently being applied in less advanced breeding programmes. This Manual was prepared to stimulate the wider use of this technology for speeding and opening up new breeding possibilities for many crops including some woody tree species. Since the construction of genetic maps using molecular markers requires the development of segregating doubled haploid populations in numerous crop species, we hope that this Manual will also help molecular biologists in establishing such mapping populations. For many years, both the Food and Agriculture Organization of the United Nations (FAO) and the International Atomic Energy Agency (IAEA) have supported and coordinated research that focuses on development of more efficient doubled haploid production methods and their applications in breeding of new varieties and basic research through their Plant Breeding and Genetics Section of the Joint F AO/IAEA Division of Nuclear Techniques in Food and Agriculture. The first F AO/IAEA scientific network (Coordinated Research Programme - CRP) dealing with doubled haploids was initiated by the Plant Breeding and Genetics Section in 1986.

This work present practical, biotechnological applications of flow cytometry techniques for the study of animal, plant and microbial cells, explaining methodologies for sample preparation, staining and analysis. It discusses cell variability in cell culture processes and shows how the quantitative analysis of heterogeneous populations aids in the biotechnological exploitation of cells.

This book provides detailed treatment of the identification and isolation of modified plant cells. No other text covers this aspect of plant cell culture technology in comparable depth. Well illustrated, lucidly written and clearly cross-referenced, the monograph emphasizes * methodology * selection of variants of agricultural and industrial interest * cell selection in relation to genetic manipulation. Plant cell biologists, molecular biologists and biotechnologists working in the field of plant cell technology will find this thorough and comprehensive book essential to their work.

Many studies in both plant and animal systems have shown that Matrix Attachment Regions (MARs) can increase expression of transgenes in whole organisms or cells in culture. MARs are AT-rich sequences of DNA that bind in vitro to the proteinacous filament-like structure within the nucleus called the nuclear matrix. In our investigation of transgenic Nicotiana tabacum NT-1 cells in culture, we have observed that transgene expression is often variegated. In other words, some cells in an isogenic population do not express the transgene, and/or other cells within the same population express the transgene at varying levels. The question was raised: Do MARs increase transgene expression by altering variegation? More specifically, do MARs increase the percentage of cells expressing the transgene, increase the magnitude of expression in expressing cells, or both? In order to address these questions, it was necessary to quantitate transgene expression variegation at the resolution of individual cells. We chose to measure Green Fluorescent Protein (GFP) expression in individual tobacco NT-1 cells by flow cytometry. In order to analyze individual cells and because NT-1 cells in culture grow as filaments, it is necessary to prepare protoplasts that can pass one at a time through the flow cell of the flow cytometer. We found that current flow cytometry methods for measuring GFP expression in plants were susceptible to debris resulting from protoplast preparations. We observed that when the plasma membrane of protoplasts is breached, GFP diffuses out into the medium, and flow cytometric measurements of these non-viable protoplasts imply they do not express GFP. This debris can overestimate the proportion of non-expressing cells in the population. In order to correct this problem, we used an approach called a dye exclusion test. Because propidium iodide enters protoplasts to stain nuclei only when the plasma membrane is breached, debris that stains with this dye can be removed from.

Targeted at beginners as well as experienced users, this handy reference explains the benefits and uses of flow cytometery in the study of plants and their genomes. Following a brief introduction that highlights general considerations when analyzing plant cells by flow cytometric methods, the book goes on to discuss examples of application in plant genetics, genomic analysis, cell cycle analysis, marine organism analysis and breeding studies. With its list of general reading and a glossary of terms, this first reference on FCM in plants fills a real gap by providing first-hand practical hints for the growing community of plant geneticists.

In recent years, the study of the plant cell cycle has become of major interest, not only to scientists working on cell division sensu strictu , but also to scientists dealing with plant hormones, development and environmental effects on growth. The book The Plant Cell Cycle is a very timely contribution to this exploding field. Outstanding contributors reviewed, not only knowledge on the most important classes of cell cycle regulators, but also summarized the various processes in which cell cycle control plays a pivotal role. The central role of the cell cycle makes this book an absolute must for plant molecular biologists.

The aim of this volume is to merge classical concepts of plant cell biology with the recent findings of molecular studies and real-world applications in a form attractive not only to specialists in the realm of fundamental research, but also to breeders and plant producers. Four sections deal with the control of development, the control of stress tolerance, the control of metabolic activity, and novel additions to the toolbox of modern plant cell biology in an exemplary and comprehensive manner and are targeted at a broad professional community. It serves as a clear example that a sustainable solution to the problems of food security must be firmly rooted in modern, continuously self re-evaluating cell-biological research. No green biotech without green cell biology. As advances in modern medicine is based on extensive knowledge of animal molecular cell biology, we need to understand the hidden laws of plant cells in order to handle crops, vegetables and forest trees. We need to exploit, not only empirically, their astounding developmental, physiological and metabolic plasticity, which allows plants to cope with environmental challenges and to restore flexible, but robust self-organisation.

This is the first book to be devoted entirely to the application and development of flow techniques in cytogenetics. It provides comprehensive information on the use of flow cytometry and sorting for chromosome classification and purification. Cytogenetics and molecular biologists will find this book an invaluable reference source. Practical details for the preparation and analysis of chromosomes using flow cytometry Flow karyotyping for sensitive rapid analysis of chromosome normality and the detection of aberrant chromosomes Flow sorting as a source of chromosome-specific DNA for gene mapping and recombinant DNA libraries Construction and current status of chromosome-specific recombinant DNA libraries