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出典(authority):フリー百科事典『ウィキペディア（Wikipedia）』「2014/10/07 08:35:05」(JST)

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Protoplasts of cells from a petunia's leaf

Protoplasts of Physcomitrella patens

Protoplast, from ancient Greek πρωτόπλαστος (prōtóplastos, "first-formed"), initially referred to the first human^{[citation needed]} or, more generally, to the first organized body of a species. In modern biology, it has several definitions:

A protoplast is a plant, bacterial or fungal cell that had its cell wall completely or partially removed using either mechanical or enzymatic means.
- 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 protoplast refers to that unit of biology which is composed of a cell's nucleus and the surrounding protoplasmic materials.

Enzymes for the preparation of protoplasts

Cell walls are made of a variety of polysaccharides. Protoplasts can be made by degrading cell walls with a mixture of the appropriate polysaccharide-degrading enzymes:

Type of cell	Enzyme
Plant cells	Cellulase, pectinase, xylanase
Gram-positive bacteria	Lysozyme (+EDTA)
Fungal cells	Chitinase

During and subsequent to digestion of the cell wall, the protoplast becomes very sensitive to osmotic stress. This means cell wall digestion and protoplast storage must be done in an isotonic solution to prevent rupture of the plasma membrane.

Uses for protoplasts

Fused protoplast (left) with chloroplasts (from a leaf cell) and coloured vacuole (from a petal).

Protoplasts can be used to study membrane biology, including the uptake of macromolecules and viruses . These are also used in somaclonal variation.

Protoplasts are widely used for DNA transformation (for making genetically modified organisms), since the cell wall would otherwise block the passage of DNA into the cell.^[1] In the case of plant cells, protoplasts may be regenerated into whole plants first by growing into a group of plant cells that develops into a callus and then by regeneration of shoots (caulogenesis) from the callus using plant tissue culture methods.^[2] Growth of protoplasts into callus and regeneration of shoots requires the proper balance of plant growth regulators in the tissue culture medium that must be customized for each species of plant. Unlike protoplasts from vascular plants, protoplasts from mosses, such as Physcomitrella patens, do not need phytohormones for regeneration, nor do they form a callus during regeneration. Instead, they regenerate directly into the filamentous protonema mimicking a germinating moss spore.^[3]

Protoplasts may also be used for plant breeding, using a technique called protoplast fusion. Protoplasts from different species are induced to fuse by using an electric field or a solution of polyethylene glycol. Originally, non–Ti-plasmid dependent DNA uptake was demonstrated in 1985 by R. Hain and A.P. Czernilofsky et al., in the article "Uptake, integration, expression and genetic transmission of a selectable chimaeric gene by plant protoplasts" using the Ca-phosphate coprecipitation technique.^[4] This technique may be used to generate somatic hybrids in tissue culture.

Additionally, protoplasts of plants expressing fluorescent proteins in certain cells maybe used for Fluorescence Activated Cell Sorting (FACS), where only cells fluorescing a chosen wavelength are retained. Among other things, this technique is used to isolate specific cell types (e.g. guard cells from leaves, pericycle cells from roots) for further investigation such as transcriptomics.

References

^ Davey MR, Anthony P, Power JB, Lowe KC (March 2005). "Plant protoplasts: status and biotechnological perspectives". Biotechnol. Adv. 23 (2): 131–71. doi:10.1016/j.biotechadv.2004.09.008. PMID 15694124.
^ Thorpe TA (October 2007). "History of plant tissue culture". Mol. Biotechnol. 37 (2): 169–80. doi:10.1007/s12033-007-0031-3. PMID 17914178.
^ 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. Protoplasma 219, 99–105.
^ R. Hain and A.P. Czernilofsky et al. (1985). "Uptake, integration, expression and genetic transmission of a selectable chimaeric gene by plant protoplasts". Mol Gen Genet 199:161–168.

English Journal

Toc33 and Toc64-III cooperate in precursor protein import into the chloroplasts of Arabidopsis thaliana.

Sommer M, Rudolf M, Tillmann B, Tripp J, Sommer MS, Schleiff E.SourceCenter of Membrane Proteomics, Cluster of Excellence 'Macromolecular Complexes' Frankfurt, Department of Biosciences, Molecular Cell Biology of Plants, Johann-Wolfgang-Goethe University Frankfurt, D-60438, Frankfurt, Germany.
Plant, cell & environment.Plant Cell Environ.2013 May;36(5):970-83. doi: 10.1111/pce.12030. Epub 2013 Jan 7.
The import of cytosolically synthesized precursor proteins into chloroplasts by the translocon at the outer envelope membrane of chloroplasts (TOC) is crucial for organelle function. The recognition of precursor proteins at the chloroplast surface precedes translocation and involves the membrane-ins
PMID 23131143

Structural characterization of an immunoenhancing heteroglycan of a hybrid mushroom (pfls1h) of Pleurotus florida and Lentinus squarrosulus (Mont.) Singer.

Sen IK, Maji PK, Behera B, Mallick P, Maiti TK, Sikdar SR, Islam SS.SourceDepartment of Chemistry and Chemical Technology, Vidyasagar University, Midnapore 721102, West Bengal, India.
Carbohydrate research.Carbohydr Res.2013 Apr 19;371:45-51. doi: 10.1016/j.carres.2013.02.004. Epub 2013 Feb 24.
An immunostimulating water-soluble heteroglycan (PS-II) was isolated from an aqueous extract of the fruit bodies of a hybrid mushroom, pfls1h produced by intergeneric protoplast fusion between Pleurotus florida and Lentinus squarrosulus (Mont.) Singer. Structural characterization of PS-II was carrie
PMID 23500959

A β-glucan from the alkaline extract of a somatic hybrid (PfloVv5FB) of Pleurotus florida and Volvariella volvacea: structural characterization and study of immunoactivation.

Maity KK, Patra S, Dey B, Bhunia SK, Mandal S, Bahera B, Maiti TK, Sikdar SR, Islam SS.SourceDepartment of Chemistry and Chemical Technology, Vidyasagar University, Midnapore 721102, West Bengal, India.
Carbohydrate research.Carbohydr Res.2013 Apr 5;370:13-8. doi: 10.1016/j.carres.2013.01.016. Epub 2013 Jan 29.
A water soluble polysaccharide isolated from the alkaline extract of the somatic hybrid mushroom (PfloVv5FB), obtained through protoplast fusion between Pleurotus florida and Volvariella volvacea strains was found to contain d-glucose only. Structural investigation was carried out using acid hydroly
PMID 23419942

Japanese Journal

スサビノリプロトプラストの生残と生長に対する抗生物質の影響

阿部真比古,藤田雄二,小林正裕 [他]
水産増殖 = Aquaculture science 63(1), 1-8, 2015-03
NAID 40020401830

An inverse relationship between allelopathic activity and salt tolerance in suspension cultures of three mangrove species, Sonneratia alba, S. caseolaris and S. ovata : development of a bioassay method for allelopathy, the protoplast co-culture method

Hasegawa Ai,Oyanagi Tomoya,Minagawa Reiko [他]
Journal of plant research 127(6), 755-761, 2014-11
NAID 40020256492

Characterization of α-1,3-glucanase isozyme from Paenibacillus glycanilyticus FH11 in a new subgroup of family 87 α-1,3-glucanase(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)

Suyotha Wasana,Yano Shigekazu,Itoh Takafumi [他],Fujimoto Hiroko,Hibi Takao,Tachiki Takashi,Wakayama Mamoru
Journal of bioscience and bioengineering 118(4), 378-385, 2014-10
… The recombinant Agl-FH1 and Agl-FH2 exhibited the same enzymatic properties as those of each wild-type enzyme, and both of the recombinants showed the activity on the protoplast formation of Schizophyllum commune mycelia. …
NAID 110009865418