Protoplasts: The protoplasm and plasma membrane of plant, fungal, bacterial or archaeon cells without the CELL WALL.Plants, Toxic: Plants or plant parts which are harmful to man or other animals.Tobacco: A plant genus of the family SOLANACEAE. Members contain NICOTINE and other biologically active chemicals; its dried leaves are used for SMOKING.Cell Wall: The outermost layer of a cell in most PLANTS; BACTERIA; FUNGI; and ALGAE. The cell wall is usually a rigid structure that lies external to the CELL MEMBRANE, and provides a protective barrier against physical or chemical agents.Bacillus megaterium: A species of bacteria whose spores vary from round to elongate. It is a common soil saprophyte.Plant Cells: Basic functional unit of plants.Plants: Multicellular, eukaryotic life forms of kingdom Plantae (sensu lato), comprising the VIRIDIPLANTAE; RHODOPHYTA; and GLAUCOPHYTA; all of which acquired chloroplasts by direct endosymbiosis of CYANOBACTERIA. They are characterized by a mainly photosynthetic mode of nutrition; essentially unlimited growth at localized regions of cell divisions (MERISTEMS); cellulose within cells providing rigidity; the absence of organs of locomotion; absence of nervous and sensory systems; and an alternation of haploid and diploid generations.Osmotic Fragility: RED BLOOD CELL sensitivity to change in OSMOTIC PRESSURE. When exposed to a hypotonic concentration of sodium in a solution, red cells take in more water, swell until the capacity of the cell membrane is exceeded, and burst.Mosaic Viruses: Viruses which produce a mottled appearance of the leaves of plants.Plant Proteins: Proteins found in plants (flowers, herbs, shrubs, trees, etc.). The concept does not include proteins found in vegetables for which VEGETABLE PROTEINS is available.Plant Leaves: Expanded structures, usually green, of vascular plants, characteristically consisting of a bladelike expansion attached to a stem, and functioning as the principal organ of photosynthesis and transpiration. (American Heritage Dictionary, 2d ed)Osmosis: Tendency of fluids (e.g., water) to move from the less concentrated to the more concentrated side of a semipermeable membrane.Arabidopsis: A plant genus of the family BRASSICACEAE that contains ARABIDOPSIS PROTEINS and MADS DOMAIN PROTEINS. The species A. thaliana is used for experiments in classical plant genetics as well as molecular genetic studies in plant physiology, biochemistry, and development.Muramidase: A basic enzyme that is present in saliva, tears, egg white, and many animal fluids. It functions as an antibacterial agent. The enzyme catalyzes the hydrolysis of 1,4-beta-linkages between N-acetylmuramic acid and N-acetyl-D-glucosamine residues in peptidoglycan and between N-acetyl-D-glucosamine residues in chitodextrin. EC An order of the ANGIOSPERMS, subclass Rosidae. Its members include some of the most known ornamental and edible plants of temperate zones including roses, apples, cherries, and peaches.Hordeum: A plant genus of the family POACEAE. The EDIBLE GRAIN, barley, is widely used as food.L Forms: Bacterial variants, unable to form a complete cell wall, which are formed in cultures by various bacteria; granules (L bodies) appear, unite, and grow into amorphous bodies which multiply and give rise to bacterial cells morphologically indistinguishable from the parent strain.Bacteriolysis: Rupture of bacterial cells due to mechanical force, chemical action, or the lytic growth of BACTERIOPHAGES.Plant Viral Movement Proteins: Viral proteins that facilitate the movement of viruses between plant cells by means of PLASMODESMATA, channels that traverse the plant cell walls.Fabaceae: The large family of plants characterized by pods. Some are edible and some cause LATHYRISM or FAVISM and other forms of poisoning. Other species yield useful materials like gums from ACACIA and various LECTINS like PHYTOHEMAGGLUTININS from PHASEOLUS. Many of them harbor NITROGEN FIXATION bacteria on their roots. Many but not all species of "beans" belong to this family.Plants, Genetically Modified: PLANTS, or their progeny, whose GENOME has been altered by GENETIC ENGINEERING.Transformation, Bacterial: The heritable modification of the properties of a competent bacterium by naked DNA from another source. The uptake of naked DNA is a naturally occuring phenomenon in some bacteria. It is often used as a GENE TRANSFER TECHNIQUE.Streptomyces: A genus of bacteria that form a nonfragmented aerial mycelium. Many species have been identified with some being pathogenic. This genus is responsible for producing a majority of the ANTI-BACTERIAL AGENTS of practical value.Transformation, Genetic: Change brought about to an organisms genetic composition by unidirectional transfer (TRANSFECTION; TRANSDUCTION, GENETIC; CONJUGATION, GENETIC, etc.) and incorporation of foreign DNA into prokaryotic or eukaryotic cells by recombination of part or all of that DNA into the cell's genome.Molecular Sequence Data: Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.Vicia faba: A plant species of the genus VICIA, family FABACEAE. The edible beans are well known but they cause FAVISM in some individuals with GLUCOSEPHOSPHATE DEHYDROGENASE DEFICIENCY. This plant contains vicine, convicine, Vicia lectins, unknown seed protein, AAP2 transport protein, and Vicia faba DNA-binding protein 1.Tobacco Mosaic Virus: The type species of TOBAMOVIRUS which causes mosaic disease of tobacco. Transmission occurs by mechanical inoculation.Sucrose: A nonreducing disaccharide composed of GLUCOSE and FRUCTOSE linked via their anomeric carbons. It is obtained commercially from SUGARCANE, sugar beet (BETA VULGARIS), and other plants and used extensively as a food and a sweetener.Schizophyllum: A genus of fleshy shelf basidiomycetous fungi, family Schizophyllaceae, order POLYPORALES, growing on woody substrata. It is pathogenic in humans.Cell Membrane: The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.Saccharomyces: A genus of ascomycetous fungi of the family Saccharomycetaceae, order SACCHAROMYCETALES.Plants, Medicinal: Plants whose roots, leaves, seeds, bark, or other constituent parts possess therapeutic, tonic, purgative, curative or other pharmacologic attributes, when administered to man or animals.Gene Expression Regulation, Plant: Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in plants.Naphthaleneacetic Acids: Naphthalene derivatives containing the -CH2CCO2H radical at the 1-position, the 2-position, or both. Compounds are used as plant growth regulators to delay sprouting, exert weed control, thin fruit, etc.Bacillus subtilis: A species of gram-positive bacteria that is a common soil and water saprophyte.Comovirus: A genus of plant viruses of the family COMOVIRIDAE in which the bipartite genome is encapsidated in separate icosahedral particles. Mosaic and mottle symptoms are characteristic, and transmission is exclusively by leaf-feeding beetles. Cowpea mosaic virus is the type species.Arabidopsis Proteins: Proteins that originate from plants species belonging to the genus ARABIDOPSIS. The most intensely studied species of Arabidopsis, Arabidopsis thaliana, is commonly used in laboratory experiments.Microscopy, Electron: Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.Closterovirus: A genus of plant viruses in the family CLOSTEROVIRIDAE containing highly flexuous filaments. Some members are important pathogens of crop plants. Natural vectors include APHIDS, whiteflies, and mealybugs. The type species is Beet yellows virus.RNA, Viral: Ribonucleic acid that makes up the genetic material of viruses.Nepovirus: A genus of polyhedral plant viruses of the family COMOVIRIDAE causing ringspots and spotting on leaves or sometimes symptomless infection. Transmission occurs by seeds, soil nematodes, or experimentally by mechanical inoculation. Tobacco ringspot virus is the type species.Bacillus: A genus of BACILLACEAE that are spore-forming, rod-shaped cells. Most species are saprophytic soil forms with only a few species being pathogenic.Culture Media: Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN.Raffinose: A trisaccharide occurring in Australian manna (from Eucalyptus spp, Myrtaceae) and in cottonseed meal.Peas: A variable annual leguminous vine (Pisum sativum) that is cultivated for its rounded smooth or wrinkled edible protein-rich seeds, the seed of the pea, and the immature pods with their included seeds. (From Webster's New Collegiate Dictionary, 1973)Caulimovirus: A genus of PLANT VIRUSES, in the family CAULIMOVIRIDAE, that are transmitted by APHIDS in a semipersistent manner. Aphid-borne transmission of some caulimoviruses requires certain virus-coded proteins termed transmission factors.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Plant Viruses: Viruses parasitic on plants higher than bacteria.Electroporation: A technique in which electric pulses of intensity in kilovolts per centimeter and of microsecond-to-millisecond duration cause a temporary loss of the semipermeability of CELL MEMBRANES, thus leading to ion leakage, escape of metabolites, and increased uptake by cells of drugs, molecular probes, and DNA.Telefacsimile: A telecommunication system combining the transmission of a document scanned at a transmitter, its reconstruction at a receiving station, and its duplication there by a copier.Genetic Vectors: DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition.Electrochemotherapy: A treatment modality that uses pulsed electrical currents to permeabilize cell membranes (ELECTROPORATION) and thereby enhance the uptake of chemotherapeutic agents, vaccines, or genes into the body's cells.SwedenTransfection: The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES.

Gibberellic acid stabilises microtubules in maize suspension cells to cold and stimulates acetylation of alpha-tubulin. (1/1558)

Gibberellic acid is known to stabilise microtubules in plant organs against depolymerisation. We have now devised a simplified cell system for studying this. Pretreatment of a maize cell suspension with gibberellic acid for just 3 h stabilised protoplast microtubules against depolymerisation on ice. In other eukaryotes, acetylation of alpha-tubulin is known to correlate with microtubule stabilisation but this is not established in plants. By isolating the polymeric tubulin fraction from maize cytoskeletons and immunoblotting with the antibody 6-11B-1, we have demonstrated that gibberellic acid stimulates the acetylation of alpha-tubulin. This is the first demonstrated link between microtubule stabilisation and tubulin acetylation in higher plants.  (+info)

Enhancer-like properties of an RNA element that modulates Tombusvirus RNA accumulation. (2/1558)

Prototypical defective interfering (DI) RNAs of the plus-strand RNA virus tomato bushy stunt virus contain four noncontiguous segments (regions I-IV) derived from the viral genome. Region I corresponds to 5'-noncoding sequence, regions II and III are derived from internal positions, and region IV represents a 3'-terminal segment. We analyzed the internally located region III in a prototypical DI RNA to understand better its role in DI RNA accumulation. Our results indicate that (1) region III is not essential for DI RNA accumulation, but molecules that lack it accumulate at significantly reduced levels ( approximately 10-fold lower), (2) region III is able to function at different positions and in opposite orientations, (3) a single copy of region III is favored over multiple copies, (4) the stimulatory effect observed on DI RNA accumulation is not due to region III-mediated RNA stabilization, (5) DI RNAs lacking region III permit the efficient accumulation of head-to-tail dimers and are less effective at suppressing helper RNA accumulation, and (6) negative-strand accumulation is also significantly depressed for DI RNAs lacking region III. Collectively, these results support a role for region III as an enhancer-like element that facilitates DI RNA replication. A scanning-type mutagenesis strategy was used to define portions of region III important for its stimulatory effect on DI RNA accumulation. Interestingly, the results revealed several differences in the requirements for activity when region III was in the forward versus the reverse orientation. In the context of the viral genome, region III was found to be essential for biological activity. This latter finding defines a critical role for this element in the reproductive cycle of the virus.  (+info)

A function for the plasmalemma grooves of a fission yeast. (3/1558)

Ultrastructural studies on regenerating protoplasts of Schizosaccharomyces pombe show that the spatial differentiation of the plasmalemma into grooves and flat areas is reflected in a functional differentiation in cell-wall synthesis. The grooves are the initial site of production of wall fibrils.  (+info)

Protection of Escherichia coli cells against extreme turgor by activation of MscS and MscL mechanosensitive channels: identification of genes required for MscS activity. (4/1558)

Mechanosensitive channels are ubiquitous amongst bacterial cells and have been proposed to have major roles in the adaptation to osmotic stress, in particular in the management of transitions from high to low osmolarity environments. Electrophysiological measurements have identified multiple channels in Escherichia coli cells. One gene, mscL, encoding a large conductance channel has previously been described, but null mutants were without well-defined phenotypes. Here, we report the characterization of a new gene family required for MscS function, YggB and KefA, which has enabled a rigorous test of the role of the channels. The channel determined by KefA does not appear to have a major role in managing the transition from high to low osmolarity. In contrast, analysis of mutants of E.coli lacking YggB and MscL shows that mechanosensitive channels are designed to open at a pressure change just below that which would cause cell disruption leading to death.  (+info)

The pro1(+) gene from Sordaria macrospora encodes a C6 zinc finger transcription factor required for fruiting body development. (5/1558)

During sexual morphogenesis, the filamentous ascomycete Sordaria macrospora differentiates into multicellular fruiting bodies called perithecia. Previously it has been shown that this developmental process is under polygenic control. To further understand the molecular mechanisms involved in fruiting body formation, we generated the protoperithecia forming mutant pro1, in which the normal development of protoperithecia into perithecia has been disrupted. We succeeded in isolating a cosmid clone from an indexed cosmid library, which was able to complement the pro1(-) mutation. Deletion analysis, followed by DNA sequencing, subsequently demonstrated that fertility was restored to the pro1 mutant by an open reading frame encoding a 689-amino-acid polypeptide, which we named PRO1. A region from this polypeptide shares significant homology with the DNA-binding domains found in fungal C6 zinc finger transcription factors, such as the GAL4 protein from yeast. However, other typical regions of C6 zinc finger proteins, such as dimerization elements, are absent in PRO1. The involvement of the pro1(+) gene in fruiting body development was further confirmed by trying to complement the mutant phenotype with in vitro mutagenized and truncated versions of the pro1 open reading frame. Southern hybridization experiments also indicated that pro1(+) homologues are present in other sexually propagating filamentous ascomycetes.  (+info)

RNA polymerase I transcription in a Brassica interspecific hybrid and its progenitors: Tests of transcription factor involvement in nucleolar dominance. (6/1558)

In interspecific hybrids or allopolyploids, often one parental set of ribosomal RNA genes is transcribed and the other is silent, an epigenetic phenomenon known as nucleolar dominance. Silencing is enforced by cytosine methylation and histone deacetylation, but the initial discrimination mechanism is unknown. One hypothesis is that a species-specific transcription factor is inactivated, thereby silencing one set of rRNA genes. Another is that dominant rRNA genes have higher binding affinities for limiting transcription factors. A third suggests that selective methylation of underdominant rRNA genes blocks transcription factor binding. We tested these hypotheses using Brassica napus (canola), an allotetraploid derived from B. rapa and B. oleracea in which only B. rapa rRNA genes are transcribed. B. oleracea and B. rapa rRNA genes were active when transfected into protoplasts of the other species, which argues against the species-specific transcription factor model. B. oleracea and B. rapa rRNA genes also competed equally for the pol I transcription machinery in vitro and in vivo. Cytosine methylation had no effect on rRNA gene transcription in vitro, which suggests that transcription factor binding was unimpaired. These data are inconsistent with the prevailing models and point to discrimination mechanisms that are likely to act at a chromosomal level.  (+info)

Long-distance RNA-RNA interactions and conserved sequence elements affect potato virus X plus-strand RNA accumulation. (7/1558)

Conserved octanucleotide sequences located upstream of two major potato virus X (PVX) subgenomic RNAs (sgRNAs), as well as elements in the 5' end of the genome, affect accumulation of sgRNA. To determine if complementarity between these sequences is important for PVX RNA accumulation, we analyzed the effects of mutations within these elements and compensatory mutations in a tobacco protoplast system and in plants. Mutations in the 5' nontranslated region (NTR mutants) that reduced complementarity resulted in lower genomic RNA (gRNA) and sgRNA levels, whereas mutations to the octanucleotide elements affected only the corresponding sgRNA levels. However, for both the NTR and octanucleotide mutants, the extent of reductions in RNA levels did not directly correlate with the degree of complementarity, suggesting that the sequences of these elements are also important. Mutants containing changes in the NTR and compensatory changes in one of the octanucleotide elements restored levels of gRNA and the other sgRNA species with an unaltered octanucleotide element to those of wild-type. Although compensatory changes significantly increased levels of the sgRNA species with the modified octanucleotide element, levels were not restored to those of wild-type. Our data indicate that long distance RNA-RNA interactions and the sequences of the interacting elements are required for PVX plus-strand RNA accumulation.  (+info)

Development of a self-cloning system for Actinomadura verrucosospora and identification of polyketide synthase genes essential for production of the angucyclic antibiotic pradimicin. (8/1558)

A self-cloning system for Actinomadura verrucosospora, a producer of the angucyclic antibiotic pradimicin A (PRM A), has been developed. The system is based on reproducible and reliable protoplasting and regeneration conditions for A. verrucosospora and a novel plasmid vector that consists of a replicon from a newly found Actinomadura plasmid and a selectable marker cloned from the Actinomadura strain. The system has an efficiency of more than 10(5) CFU/microgram of DNA. Using this system, we have cloned and identified the polyketide synthase (PKS) genes essential for PRM A biosynthesis from A. verrucosospora. Nucleotide sequence analysis of the 3.5-kb SalI-SphI fragment showed that ketosynthase subunits (open reading frame 1 [ORF1] and ORF2) of the essential PKS genes have strong similarities (59 to 89%) to those for angucyclic antibiotic biosynthesis.  (+info)

  • Conclusion] The preparation system has been successfully established, which lays foundation for establishing protoplast regeneration system from N. fordii leaves. (
  • For these reasons, most researchers now prefer the enzymatic method, except in cases where the enzymes can have a damaging effect on the resulting protoplasts. (
  • DNA-free gene editing can be achieved by using purified Cas9 enzymes with gRNA and transfecting them directly into your cells or protoplasts of interest. (
  • In other cases, the protoplasts were diluted into molten soft agar at 45 1C containing either LB medium and 0.5 M sucrose or M9 medium, 0.5 M sucrose, and either Leu (100 mg/ml) or Arg (100 mg/ml), depending on the nutritional requirements of the auxotrophic strains used, and then poured onto soft agar plates of the same composition. (
  • Barbier-Brygoo H., Ephritikhine G., Klämbt D., Guern J. (1990) The Sensitivity of Plant Protoplasts to Auxin is Likely Modulated by the Number of Receptors at the Plasmalemma. (
  • In some cases, protoplasts were resus- pended in LB medium containing 0.5 M sucrose and spread onto soft agar plates containing LB medium and 0.5 M sucrose with autoclaved plastic spreaders. (
  • In the past, protoplasts were isolated using mechanical methods which were labor-intensive, tedious and inefficient. (
  • Method] Taking the yield and survival rate of protoplast as indices, the effects of different factors, including pretreatment, enzymolysis method, enzymolysis temperature, centrifugation speed, illumination, and sieve mesh number were studied for the separation and purification of protoplast from N. fordii leaves. (
  • protoplast ( energid ) The living unit of a cell, consisting of the nucleus and cytoplasm bounded by the plasma membrane. (
  • This means cell wall digestion and protoplast storage must be done in an isotonic solution to prevent rupture of the plasma membrane. (
  • A.R.D. Taylor and J.L. Hall, An ultrastructural comparison of lanthanum and silicotungstic acid/chromic acid as plasma membrane stains of isolated protoplasts, Plant Sci. (
  • The freezing tolerance of protoplasts isolated from nonacclimated rye leaves (Secale cereale L. cv Puma) was significantly altered by using a pH-induced protoplastliposome fusion technique to modify the lipid composition of the plasma membrane. (
  • In control nonacclimated protoplasts, osmotic contraction resulted in endocytotic vesiculation ofthe plasma membrane which was irreversible and resulted in lysis during osmotic expansion after melting of the suspending medium. (
  • Normally, isolated protoplasts do not fuse with each other because the surface of the isolated protoplast carries negative charge (-10 to -30 mV) around the outside of plasma membrane and, thus, there is a strong tendency for pro-toplasts to repel one another due to their same charges. (
  • The development of defined protoplast transient expression systems for high-throughput screening and systematic characterization of gene functions has greatly contributed to elucidating plant signal transduction pathways, in combination with genetic, genomic, and transgenic approaches. (
  • In barley ( Hordeum vulgare ) aleurone protoplasts, the endogenous α-amylase gene is regulated by ABA and GA in parallel to what is observed in seeds ( Jacobsen and Beach, 1985 ). (
  • set the foundation to use protoplasts to study gene regulation and signal transduction in plant cells. (
  • Chemical methods for direct gene transfer to protoplasts. (
  • To obtain stable transformants we used two methods of protoplast culture after PEG-mediated gene transfer. (
  • The stimulation of GUS activity in protoplasts cotransformed with vectors containing FRT inactivated gusA gene and a chimeric FLP gene depended on both the expression of the FLP recombinase and the presence and structure of the FRT sites. (
  • Possibilities of using protoplast fusion for gene mapping and strain improvement are suggested. (
  • Dominant selectable markers, namely, kanamycin resistance ( nptII ) and hygromycin B resistance ( hpt ) genes had been previously integrated in the nuclear genomes of the otherwise almost fully isogenic parental strains using direct gene transfer to protoplasts. (
  • Transformed protoplasts have been used to assay gene function using metabolic profiling, to explore protein-DNA interactions using chromatin immunoprecipitation (ChIP), and to assay the activation of reporter constructs by transcription factors. (
  • The rolB Gene of Agrobacterium rhizogenes Does Not Increase the Auxin Sensitivity of Tobacco Protoplasts by Modifying the Intracellular Auxin Concentration. (
  • The genetic requirement for t 0 rescuer protoplasts was that they were spo + for the gene to be complemented. (
  • There were some arguments or concerns in using protoplasts in gene function studies. (
  • Protoplasts can be made by degrading cell walls with a mixture of the appropriate polysaccharide-degrading enzymes: During and subsequent to digestion of the cell wall, the protoplast becomes very sensitive to osmotic stress. (
  • Cultivation in liquid medium, followed by mycelium treatment with Novozym 234 in the presence of KCl 0.7M as osmotic stabilizer, produced 5.05 x 10 6 to 1.15 x 10 7 x mL -1 protoplasts. (
  • The permeability of isolated protoplasts was assessed by the relative ability of solutes to effect osmotic stabilization. (
  • An yield improvement in our laboratory will be the result of new investigations on initial density of protoplast, osmotic pressure, composition of culture media and the utility of agar adjunction at the first step of culture. (
  • In nonacclimated protoplasts fused with mono- or diunsaturated species of PtdCho, osmotic contraction resulted in the reversible formation of exocytotic extrusions of the plasma membraneas normally occurs in protoplasts isolated from cold-acclimated leaves (acclimated protoplasts). (
  • The influence of different cell age, enzymolysis time, temperature and osmotic pressure stabilizer on productive rate of protoplast from Lentinula edodes were discussed in this paper. (
  • It is the best conditions of preparing protoplast with Lentinula edodes hyphae that cell age was 6 days, That enzymolysis time was 3 hours, that enzymolysis temperature was 34℃ and that concentration of mannitol as osmotic pressure stabilizer was 0.6mol/l. (
  • An osmotic stabilizer, 0.6 M sucrose in 20 mM phosphate buffer pH 5.8, gave the highest yield of protoplasts. (
  • Whereas the reversion of ring-lacking protoplasts coincided with the polarization of the dotted F-actin pattern, the ring-containing protoplasts became furrowed as the F-actin rings constricted. (
  • Protoplasts from different species are induced to fuse by using an electric field or a solution of polyethylene glycol. (
  • Fusing protoplasts from two different species into a single cell can produce a hybrid plant. (
  • We chose cabbage, rapeseed and Chinese cabbage as species representatives and introduced RNPs into their protoplasts with PEG 4000. (
  • Curing of cryptic molecules from multiple plasmid complements by protoplast regeneration may prove to be generally valuable in lactic streptococci and other gram-positive species. (
  • If traditional methods are usually used with witloof chicory tissues to multiply interesting genotypes, protoplasts allow new possibilities as species amelioration and selection. (
  • Proteins annotated to have RNA-binding capacity were overrepresented in the obtained medium scale mRNA-bound proteome, indicating the specificity of the method and providing in vivo UV crosslinking experimental evidence for several candidate RBPs from leaf mesophyll protoplasts. (
  • Protoplasts are fungal, plant or gram-positive bacterial cells without a cell wall. (
  • Protoplast, from ancient Greek πρωτόπλαστος (prōtóplastos, "first-formed"), in biology, it was proposed by Hanstein (1880) to refer to the entire cell, excluding the cell wall, but currently has several definitions: a plant, bacterial or fungal cell that had its cell wall completely or partially removed using either mechanical or enzymatic means. (
  • A further differentiation can be made for bacteria: protoplasts: have their cell wall entirely removed and are derived from gram + (gram-positive) spheroplasts: have their cell wall only partially removed and are gram - (gram-negative) more generally, a unit of biology which is composed of a cell's nucleus and the surrounding protoplasmic materials. (
  • Additionally, protoplasts of plants expressing fluorescent proteins in certain cells may be used for Fluorescence Activated Cell Sorting (FACS), where only cells fluorescing a selected wavelength are retained. (
  • Protoplasts are spherical naked plant cells produced by the removal of the cell wall with digestive enzymes (Fig. 1). (
  • Plant protoplasts can be cultured in defined media and will form a new cell wall, divide, and in many cases regenerate complete plants (e.g. 1, 2, 3, 4). (
  • Binding, H., 1974a, Cell cluster formation by leaf protoplasts from axenic cultures of haploid Petunia hybrida L. (
  • versatilis than in S. pombe protoplasts and it was blocked when the reversion was inhibited by Novozyme 234 indicating that the cell wall formation is essential for the F-actin ring constriction. (
  • Plant protoplasts show physiological perceptions and responses to hormones, metabolites, environmental cues, and pathogen-derived elicitors, similar to cell-autonomous responses in intact tissues and plants. (
  • Compared with cell culture lines, the use of fresh tissues as protoplast sources offers unique advantages. (
  • A protoplast is a plant cell that has had its tough outer cell wall removed by chemical treatment. (
  • Selection of kanamycin-resistant cell lines after treatment of BMS protoplasts with PEG and pCaMVNeo (50ug. (
  • Altogether, the best estimate of the limiting equivalent porosity of the protoplast was 0.4 to 0.6 nm in radius and of the cell wall, 1.1 nm. (
  • Protoplast formation, fusion, and cell regeneration have been achieved with mutant strains of Actinoplanes brasiliensis. (
  • The plant protoplast provides an excellent means not only for characterizing cell wall proteins, but also for visualizing the dynamics of cell wall regeneration, during which cell wall proteins are secreted. (
  • This review deals with sub-proteomic approaches to the plant cell wall through the use of protoplasts, a methodology that will provide the basis for further exploration of cell wall proteins and cell wall dynamics. (
  • Large amount of intact and clean protoplast with very few cell debris was obtained. (
  • Fungicidal action was evaluated against Trametes versicolor and Postia placenta using both protoplasts and cell fragments. (
  • The giant multi-nucleate cells of the green alga, Boergesenia forbesii, undergo segregative cell division upon wounding such that small protoplasts are formed within 2 hours. (
  • Regeneration of fertile barley plants from mechanically isolated protoplasts of the fertilized egg cell Plant Cell 6: 531-543. (
  • A monoclonal antibody that defines a surface antigen on Candida albicans hyphae cross-reacts with yeast cell protoplasts. (
  • In a Western blot, MAb 1.183 showed reactivity with a 20-kDa protein in the sodium dodecyl sulfate extract from protoplasts, whereas no reactivity was found with cell wall material obtained from yeast cells. (
  • To clarify the involvement of actin in the formation of the yeast cell wall, reverting protoplasts of Schizosaccharomyces pombe were used as a simple model system. (
  • A close spatial as well as temporal relationship between actin and cell wall formation was observed in protoplast reversion. (
  • That is, the site of actin 'dots' in the reverting protoplasts coincided with the site of new wall formation and the timing of rearrangement of actin coincided with the initiation of cell wall formation and with the timing of cell wall expansion. (
  • Treatment of reverting protoplasts with cytochalasin D (CD) further clarified the close relationship between actin and cell wall organization. (
  • A low dose of CD caused weakly stained unlocalized actin, which induced grossly aberrant cell wall deposition as well as substantial changes in the morphology of the reverting protoplasts. (
  • These results demonstrated that actin is associated with initiation of cell wall formation, the proper deposition of cell wall materials, and maintaining the normal morphology of reverting protoplasts. (
  • The sub-protoplasts were spherical, so their two-dimensional areas (πr 2 ) were summed for each cell by the computer and multiplied by 4 to give the total surface area. (
  • Protoplast culture and electric cell fusion of cruciferous vegetables were studied. (
  • On the other hand, pollen culture was not formed colony, since protoplasts from pollen grains showed only 2 or 3 cell divisions. (
  • Because the visualization of MTs by immunofluorescence is technically difficult in intact hyphae of A. nidulans, we developed a method for removing the cell wall under conditions that do not perturb cell physiology, as evidenced by the fact that the resulting protoplasts undergo nuclear division at a normal rate and that cell cycle mutant phenotypes are expressed at restrictive temperature. (
  • Both protoplasts and spheroplasts refer to altered forms of plant, bacterial or fungal cells from which the cell wall has been partially or completely removed. (
  • When used in reference to bacterial cells, protoplasts may also refer to the spherical shape assumed by gram-positive bacteria while spheroplasts refer to the spherical shape assumed by gram-negative bacteria upon partial or complete removal of the cell wall. (
  • Since cell walls are composed of a variety of polysaccharides, viable protoplasts can be prepared by weakening the primary stress-bearing layer of the cell wall (peptidoglycan) using the appropriate enzymes. (
  • For plant cells, cellulase, pectinase, and xylanase can be used to break down the cell walls while lysozome (+EDTA) can be used to produce protoplasts from gram-positive bacteria. (
  • Levels of abscisis acid (ABA) were determined in isolated guard cell (GCP) and mesophyll cell (MCP) protoplasts of Vicia faba L. in relation to water stress. (
  • Cell wall oligogalacturonides increase cytosolic free calcium in carrot protoplasts. (
  • Protoplast fusion is the joining of two plant cells together to form one cell with multiple nuclei. (
  • Gunilla) protoplasts and after illumination for 10 min at high and very low CO2 in the presence of respiratory and photorespiratory inhibitors. (
  • Böttcher UF, Aviv D, Galun E (1989) Complementation between protoplasts treated with either of two metabolic inhibitors results in somatic hybrid plants. (
  • The manifestation of regular Ca 2+ oscillations depends on the concentrations of oat protoplasts in suspension that suggests the propagation of a signal between the cells and synchronization of their light- induced Ca 2+ oscillations response. (
  • Benomyl at concentrations up to 50 ppm did not exhibit toxic effects in suspension or protoplast cultures if dissolved by autoclaving or boiling. (
  • A low-K(m) component was found in protoplast-derived cells (K(m) = 45 +/- 5 micromolar) as well as in suspension-cultured cells (K(m) = 84 +/- 21 micromolar). (
  • It was present in protoplast-derived cells but not in suspension-cultured cells, and had the same magnitude in wild-type and mutant cells. (
  • The reduced V(max) of the low-K(m) component in the Val(r)-2 mutant, and the differential expression of the other two components in suspension-cultured cells and protoplast-derived cells indicate that the kinetically distinguishable components represent physically distinct transport systems. (
  • Splenocytes, derived from mice that had been immunized with protoplasts prepared from suspension cultures of root cells of Glycine max (L.) Merr. (
  • A similar yield of protoplasts (1-2 x 10(5) ml-1 after 16 h) was produced by using either enzymatic system. (
  • We have demonstrated that a yeast FLP/FRT site-specific recombination system functions in maize and rice protoplasts. (
  • When exponentially growing yeast cells were treated sequentially with EDTA, beta-mercaptoethanol, and Zymolase to form protoplasts, a specific immunofluorescence signal was obtained with MAb 1.183. (
  • In summary, we demonstrated the suitability of RNP transfection into B. oleracea and B. rapa protoplasts for high-efficiency indel induction of two endogenous genes. (
  • The loss of five different plasmids, including small multicopy molecules, was readily detected in Streptococcus lactis 712 by screening lysates of random protoplast regenerants on agarose gels. (
  • S.C. Bhatla, Justine Kiessling, Ralf Reski (2002): of polarity induction by cytochemical localization of phenylalkylamine-binding receptors in regenerating protoplasts of the moss Physcomitrella patens. (
  • J. Burgess, P.J. Linstead and V.E.L. Fisher, Studies on higher plant protoplasts by scanning electron microscopy, Micron 8:113-122 (1977). (
  • In scanning electron micrographs, the morphology of the extrusions of nonacclimated protoplasts fused with [LintdCho was virtually indistinguishable from that of the extrusions formed in acclimated protoplasts. (
  • Scanning electron microscopy revealed the presence of a fibrillar net structure on the surface of non-treated control reverting protoplasts. (
  • The in planta analysis of elements involved in phytohormone signaling can be achieved through transient expression in mesophyll protoplasts, which are a fast and versatile alternative to generating plant lines that stably express a transgene. (
  • Preparation and Purification of Protoplast from Nervilia fordii (Hance) Schltr. (
  • 摘要】 [Objective] To set up protoplast preparation and purification method from Nervilia fordii (Hance) Schltr. (
  • Method] Taking the yield and survival rate of protoplast as indices, the effects of different factors, including pretreatment, enzymolysis method, enzymolysis temperature, centrifugation speed, illumination, and sieve mesh number were studied for the separation and purification of protoplast from N. fordii leaves. (
  • Therefore, in order to examine those changes in respiratory activity peculiar to the host, attempts were made to isolate protoplasts from infected tissues, and from healthy controls. (
Genetic Manipulations with Plant Material | SpringerLink
Genetic Manipulations with Plant Material | SpringerLink (
Difference Between Spheroplasts and Protoplasts
Difference Between Spheroplasts and Protoplasts (
Plant Protoplasts: An Experimental System for Cell Biologists | SpringerLink
Plant Protoplasts: An Experimental System for Cell Biologists | SpringerLink (
Frontiers | TOR-Dependent and -Independent Pathways Regulate Autophagy in Arabidopsis thaliana | Plant Science
Frontiers | TOR-Dependent and -Independent Pathways Regulate Autophagy in Arabidopsis thaliana | Plant Science (
Techniques of Somatic Cell Hybridization by Fusion of Protoplasts | SpringerLink
Techniques of Somatic Cell Hybridization by Fusion of Protoplasts | SpringerLink (
May 1982 - Scientific American
May 1982 - Scientific American (
A method for high-frequency intergeneric fusion of plant protoplasts | SpringerLink
A method for high-frequency intergeneric fusion of plant protoplasts | SpringerLink (
IJMS  | Free Full-Text | Efficient Genome Editing Using CRISPR/Cas9 Technology in Chicory | HTML
IJMS | Free Full-Text | Efficient Genome Editing Using CRISPR/Cas9 Technology in Chicory | HTML (
Structural insights into Arabidopsis ethylene response factor 96 with an extended N-terminal binding to GCC box | SpringerLink
Structural insights into Arabidopsis ethylene response factor 96 with an extended N-terminal binding to GCC box | SpringerLink (
Tobacco plant protoplasts, SEM - Stock Image B060/0065 - Science Photo Library
Tobacco plant protoplasts, SEM - Stock Image B060/0065 - Science Photo Library (
Frontiers | Progress in Somatic Embryogenesis of Japanese Pines | Plant Science
Frontiers | Progress in Somatic Embryogenesis of Japanese Pines | Plant Science (
Plus it
Plus it (
Plus it
Plus it (
Reagents<... (
Holzforschung (
Plus it
Plus it (
General Properties of Isolated Protoplasts and Uptake of Foreign Genetic Material | SpringerLink
General Properties of Isolated Protoplasts and Uptake of Foreign Genetic Material | SpringerLink (
Plus it
Plus it (
Plant Cell Culture Protocols | Springer for Research & Development
Plant Cell Culture Protocols | Springer for Research & Development (
Frontiers | Dehydrins Impart Protection against Oxidative Stress in Transgenic Tobacco Plants | Plant Science
Frontiers | Dehydrins Impart Protection against Oxidative Stress in Transgenic Tobacco Plants | Plant Science (
Plus it
Plus it (
Balance between Cytosolic and Chloroplast Translation Affects Leaf Variegation | Plant Physiology
Balance between Cytosolic and Chloroplast Translation Affects Leaf Variegation | Plant Physiology (
Protoplast of moss, Physcomitrella - Stock Image - B060/0015 - Science Photo Library
Protoplast of moss, Physcomitrella - Stock Image - B060/0015 - Science Photo Library (
Direct Transfer of Synthetic Double-Stranded RNA into Protoplasts of Arabidopsis thaliana | SpringerLink
Direct Transfer of Synthetic Double-Stranded RNA into Protoplasts of Arabidopsis thaliana | SpringerLink (
The Magnesium-Chelatase H Subunit Binds Abscisic Acid and Functions in Abscisic Acid Signaling: New Evidence in Arabidopsis |...
The Magnesium-Chelatase H Subunit Binds Abscisic Acid and Functions in Abscisic Acid Signaling: New Evidence in Arabidopsis |... (
Spheroplast - Wikipedia
Spheroplast - Wikipedia (
HDA9-PWR-HOS15 Is a Core Histone Deacetylase Complex Regulating Transcription and Development | Plant Physiology
HDA9-PWR-HOS15 Is a Core Histone Deacetylase Complex Regulating Transcription and Development | Plant Physiology (
Search Results - AccessScience from McGraw-Hill Education
Search Results - AccessScience from McGraw-Hill Education (
Plus it
Plus it (
Chloroplasts in protoplast of tobacco - Stock Image - B100/0037 - Science Photo Library
Chloroplasts in protoplast of tobacco - Stock Image - B100/0037 - Science Photo Library (
Category:Cell biology - Wikimedia Commons
Category:Cell biology - Wikimedia Commons (
Category:Cell biology - Wikimedia Commons
Category:Cell biology - Wikimedia Commons (