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Protoplast

About: Protoplast is a research topic. Over the lifetime, 5474 publications have been published within this topic receiving 122468 citations.


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Journal ArticleDOI
15 Jun 1957-Nature
TL;DR: Nečas3 showed that spontaneously autolysing yeast gave rise to a small extent to structures which, superficially at least, resembled protoplasts.
Abstract: A NEW approach to the structure and functions of the cell surface of certain bacteria was revealed when Weibull1 showed that, in the presence of sucrose, lysozyme dissolves the cell-wall, leaving the protoplast essentially intact. Various attempts have since been made to isolate protoplasts from bacteria normally insensitive to lysozyme2 and also from yeast. Thus Necas3 showed that spontaneously autolysing yeast gave rise to a small extent to structures which, superficially at least, resembled protoplasts.

215 citations

Journal ArticleDOI
TL;DR: Protoplasts isolated from cultured rice cells of an A-58 cytoplasmic male sterile line (A-58 MS)(Oryza sativa L.) were used to investigate the regeneration of rice plants.
Abstract: Protoplasts isolated from cultured rice cells of an A-58 cytoplasmic male sterile line (A-58 MS)(Oryza sativa L.) were used to investigate the regeneration of rice plants. A cultured cell line (T3) of A-58 MS with a high growth rate and dense cytoplasm was selected. About 10% of the protoplasts prepared from this established cell line plated in RY-2 (a new medium) formed colonies. The calli formed shoots and roots in the regeneration medium and developed into whole plants.

215 citations

Journal ArticleDOI
TL;DR: Isolated egg cells of maize showed protoplasmic streaming during 22 days of culture, but they did not divide, but after fusion of the sperm with the egg cells, these fused cells did develop, with a mean division frequency of 83%, and grew to multicellular structures.
Abstract: Electrofusion-mediated in vitro fertilization of maize using single sperm and egg cells was performed. Sperm cells were released from pollen grains after rupture of the latter by osmotic shock in the fusion medium (0.55 M mannitol). Egg cells were isolated by enzyme treatment (pectinase, pectolyase, hemicellulase, and cellulase) followed by mechanical isolation. The conditions generally used for the electrical fusion of protoplasts of somatic cells were also applied to the protoplasts of gametic cells of maize. Electrofusion was performed with single pairs of gametes under microscopic observation. The mean fusion frequency was 79%. Isolated egg cells of maize showed protoplasmic streaming during 22 days of culture, but they did not divide. However, after fusion of the sperm with the egg cells, these fused cells did develop, with a mean division frequency of 83%, and grew to multicellular structures. Egg cells and fusion products were cultivated with a maize feeder-cell system.

212 citations

Journal ArticleDOI
TL;DR: This protocol details an in vivo transient expression system to study the subcellular localization and dynamic associations of plant proteins using protoplasts freshly prepared from Arabidopsis or tobacco BY-2 suspension cultured cells.
Abstract: Transient expression of fluorescent fusion proteins in plant cells has dramatically facilitated our study of newly identified genes and proteins. This protocol details an in vivo transient expression system to study the subcellular localization and dynamic associations of plant proteins using protoplasts freshly prepared from Arabidopsis or tobacco BY-2 suspension cultured cells. The method relies on the transformation of DNA constructs into protoplasts via electroporation. The whole protocol is comprised of three major stages: protoplast generation and purification, transformation of DNA into protoplasts via electroporation and incubation of protoplasts for protein analysis. Similar to stably transformed cell lines, transformed protoplasts are compatible with protein localization studies, pharmaceutical drug treatment and western blot analysis. This protocol can be completed within 11-24 h from protoplast production to protein detection.

210 citations

Journal Article
TL;DR: Rapeseed plants have been regenerated after fusion between protoplasts bearing cytoplasms of different genera and transfer of chloroplasts has been confirmed by restriction cp DNA analysis and two dimensional thylakoïd protein electrophoresis.
Abstract: SummaryRapeseed plants have been regenerated after fusion between protoplasts bearing cytoplasms of different genera. Cybrids combine, in a first experiment Brassica napus chloroplasts and a cytoplasmic male sterility (cms) trait coming from Raphanus sativus, in a second experiment chloroplasts of a triazine resistant Brassica campestris and cms trait from Raphanus sativus. Transfer of chloroplasts has been confirmed by restriction cp DNA analysis and two dimensional thylakoïd protein electrophoresis. These plants may be very useful for Brassica hybrid seed production.

210 citations


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Performance
Metrics
No. of papers in the topic in previous years
YearPapers
202358
2022153
202160
202060
201978
201855