Protoplast

This detailed volume collects protocols from scientists who are actively engaged in developing or using protoplast technology. The book begins with chapters that focus on basic protoplast techniques and their utilities, such as protocols on protoplast isolation, transfection, and regeneration, as well as examples of how to use protoplasts for genome editing and gene function analysis in a number of major crop or model plant species. It continues by exploring protoplast automation, large scale functional genomics, and synthetic biology. Written for the highly successful Methods in Molecular Biology series, 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 practical, Protoplast Technology: Methods and Protocols aims to inspire the new generation of researchers to further improve their protocols and apply this technology to accelerate the field of plant genomic study.

Plant biotechnology has created unprecedented opportunities for the manipulation of biological systems of plants. To understand biotechnology, it is essential to know the basic aspects of genes and their organization in the genome of plant cells. This text on the subject is aimed at students.

Proceedings of the 7th International Protoplast Symposium, Wageningen, The Netherlands, December 6-11, 1987

In continuation of Volumes 8, 9, 22, and 23, this new volume deals with the regeneration of plants from isolated protoplasts and genetic transformation in various species of Actinidia, Allocasuarina, Anthurium, Antirrhinum, Asparagus, Beta, Brassica, Carica, Casuarina, Cyphomandra, Eucalyptus, Ipomoea, Larix, Limonium, Liriodendron, Malus, Musa, Physcomitrella, Physalis, Picea, Rosa, Tagetes, Triticum, and Ulmus. These studies reflect the far-reaching implications of protoplast technology in genetic engineering of plants. The book contains a wealth of useful information for advanced students, teachers, and researchers in the field of plant tissue culture, molecular biology, genetic engineering, plant breeding, and general biotechnology.

Plant Protoplasts covers the techniques involved with, and uses of, protoplast technology. The book discusses isolation, fusion, and culture of higher plant protoplasts, lower plant protoplasts, and blue-green algal protoplast. The text also describes the production of haploid protoplasts from developing pollen grains; the use of protoplasts in mutant selection schemes, and the development of protoplast systems for use with monocotyledonous plants. The book will be invaluable to plant technologists, botanists, biochemists, research workers as well as advanced students interested in gaining a background knowledge of the field.

The present book thus emphasizes the importance of plant protoplasts for fundamental research. This book is intended to be used b senior undergraduates, graduate students, and research scientists in plant biology.

Isolated plant protoplasts are 'naked' cells that have had their cell wall removed either by mechanical action or by enzymic digestion. As a result of wall removal the only barrier that exists between the cell protoplasm and the external environment is the plasma membrane. The removal of the cell wall has drastic osmotic consequences for the isolated cell. Rather like animal cells, the isolated protoplasts must be maintained in an osmotically balanced (isotonic) nutrient medium or they will burst. As will be seen later, the isolation of plant protoplasts is not a particularly new event; as early as the 1890s scientists were isolating protoplasts mechanically and studying effects such as protoplast streaming. What has brought protoplast technology to the forefront of plant biology is the potential application of these structures in non-traditional plant improvement methods. Extreme care must be exercised by the reader, however, in be lieving some of the claims that have been put forward for the use of protoplasts in improvement programmes. In the 20 years since the first experiments in routine enzyme isolation of large numbers of protoplasts no real improvement has been obtained in a commercial crop plant. This does not mean that improvement is impossible; it simply indicates that it may be a longer term objective than some people would like to admit.

Twenty-seven chapters deal with the regeneration of plants from protoplasts and genetic transformation in various species of Agrostis, Allium, Anthriscus, Asparagus, Avena, Boehmeria, Carthamus, Coffea, Funaria, Geranium, Ginkgo, Gladiolus, Helianthus, Hordeum, Lilium, Lithospermum, Mentha, Panax, Papaver, Passiflora, Petunia, Physocomitrella, Pinus, Poa, Populus, Rubus, Saintpaulia, and Swertia. These studies reflect the far-reaching implications of protoplast technology in genetic engineering of plants. This volume is of special interest to advanced students, teachers, and research scientists in the field of plant tissue culture, molecular biology, genetic engineering, plant breeding, and general plant biotechnology.