Microbial And Plant Protoplasts

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.

Recent developments in the culture and regeneration of plant protoplasts; Protoplast culture and plant regeneration of cereals and other recalcitrant crops; Protoplasts and the isolation of plant mutants; Protoplasts and variation from culture; Systems for plant protoplast transformation; Recent developments in plant protoplast fusion and selection technology; Somatic hybridization by plant protoplast fusion; The segregation of organelles and cytoplasmic traits in higher plant somatic fusion hybrids; The first mitotic cycle of mesophyll protoplasts; Protoplasts for studies of the plasma membrane and associated cell organelles; The use of protoplasts in plant virus research; Applications of protoplast technology to agriculture; Organelle transfer, sorting out, recombination; Plant protoplasts as tools for physiological studies; Genetic transformation; Protoplasts as tools in pathology, virology and plant-microbe interactions; Current questions of gene transfer via protoplast fusion in microorganisms (Opening adress); Advances in protoplast fusion and transformation in Streptomyces; Chromosome interactions and expression in fused Bacillus protoplasts; Yeast strain improvement by protoplast fusion and transformation; Protoplasts of filamentous fungi in genetics and metabolite production; Interspecific somatic hybridisation in Aspergillus; Synthesis and assembly of wall polymers on regenerating yeast protoplasts; The theory and practical applications of liposome-protoplast interactions; Transport of nutrients in yeast protoplast; Protoplasts and spheroplasts of gram-negative bacteria - with special emphasis on proteus mirabilis; Characteristic properties and biological significance of stable protoplast type L-forms; Advances in microbial protoplasts (Bacillus licheniformis lactamase and the protoplast surface); Fungal protoplasts as genetic tools; Non-complementing diploids: prokaryotic microorganism protoplast fusion and chromosome inactivation.

In the series of International Protoplast Symposia the Symposium of 1987 was held in Wageningen (The Netherlands). Earlier Symposia took place in Jena (DDR) 1963, Brno (CSSR) 1967, Salamanca (Spain) 1971, Nottingham (UK) 1975, Szeged (Hungary) 1979 and Basel (Switzerland) 1983. This 7th International Protoplast Symposium was organized by K.J. Puite (Secretary), J.J.M. Dons (Treasurer), H.J.Huizing and E.J.L. Hotke-Staal (Local Organizers), the first three persons being scientists, respectively, from the Research Institute Ital, the Institute for horticultural plant breeding IVT and the Foundation for agricultural plant breeding SVP at Wageningen. Scientific Advisers of the Symposium were A. J. Kool, M. Koornneef and F.A. Krens. The International Agricultural Centre lAC served as the Symposium location. The Organizing Committee decided that the scientific programme of the Symposium should be mainly focussed on protoplast technology of relevance to plant breeding. Therefore research on microbial protoplasts and on secondary metabolites was not included. About 250 scientists from 27 different countries were welcomed at the meeting. Speakers at Symposium Sessions and authors of Poster contributions were asked to hand over their manuscripts for the Symposium Proceedings already at the meeting, permit ting early publication of the Proceedings. These manuscripts give the state of the art of the protoplast research and illustrate the progress since the last Protoplast Symposium.

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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.