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By P. E. Pilet (auth.), Professor Dr. Paul-Emile Pilet (eds.)

ISBN-10: 3642701442

ISBN-13: 9783642701443

ISBN-10: 3642701469

ISBN-13: 9783642701467

The thought for the p~esent ebook arose from a 3-day seminar which I geared up in March 1984 for younger examine staff in plant body structure. members got here from a number of universities of the French-speaking a part of Switzerland and audio system from Basel, Mtinchen, Nottingham, Perpignan, Regensburg, Sheffield, Toulouse, Yale, ZUrich ... and Lausannne. The topic of the seminar used to be selected from the variety of study fields of our Institute. in the meantime, feeling it used to be very important to keep in mind that a few of our hearers weren't experts within the selected subject, i wanted to professional­ vide an issue that may be of clinical, methodological and epistemologi­ cal curiosity. The serious research of the structural and practical features of plant protoplasts precisely met those standards. There exists abundant fabric for dialogue of the recommendations of protoplast training, tools utilized in morphological, organic and biochemical reports, and for the comparability of protoplasts with the cells from which they're obtained.

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Planta (Berl) 143:33-39 Shepard JF (1981) The isolation, culture, and regeneration of plant protoplasts. Course on plant molecular biology. Cold Spring Harbor Laboratory, USA Stadelman EJ, Kinzel H (1972) Vital staining of plant cells. In: Prescott DM (ed) Methods in cell physiology, vol 5. Academic, New York Vatsya B, Bhaskaran S (1981) Production of subprotoplasts in Brassica oleracea var. Capitata a function of osmolarity of the media. Plant Sci Lett 23:277-282 Wallin A, Glimelius K, Eriksson T (1978) Enucleation of plant protoplasts by cytochalasin B.

This is not always the case (Quail 1979, Parish 1983). Mophometric quantitation (Morn~ and Buckhout 1979) may be useful in the determination of membrane contamination of isolated plasma membrane "sheets" fractions. Electron microscopy of thin section of fraction F3 prepared after membrane stabilization with myeloma protein J539 showed two main contaminants: protein bodies and a few starch granules. These protein bodies are frequently observed at the periphery of protoplasts in close proximity to the plasma membrane.

Isolation of Plasma Membrane from Ryegrass 39 Table 1. Methods used to label the plasma membrane of intact protoplasts Labelling reagent/ligand Target groups Label Labelpermeates ling ofLm. m. 1251 and Iodgen Tyrosine, histidine Yes No a Fraker and Speck 1978 125 I and lactoperoxidase/ glucose oxidase Tyrosine, histidine Yes No a Suzuki et al. 1980 Galactose oxidase and [3H]-borohydride Terminal galactose N-acetyl galactosamine Yes No a Gahmberg and Hakamori 1973 Amino groups, tyrosine histidine Yes No a Gailbraith and Northcote 1977 [12SI]-iodosulphanilic acid Amino groups, tyrosine histidine Yes No a Perlin and Spanswick 1980 [12SI]_myeloma protein JS39 1,6-{3-D-galactopyranosyl No No Schibeci et al.

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The Physiological Properties of Plant Protoplasts by P. E. Pilet (auth.), Professor Dr. Paul-Emile Pilet (eds.)

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