3/28/2019 Protoplast - Wikipedia

Protoplast
Protoplast, from ancient Greek πρωτόπλ ασ τος (prōtóplastos, "first-
formed"), is a biological term proposed by Hanstein in 1880 to refer to the
entire cell, excluding the cell wall, [1 ][2 ] but currently has sev eral
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)
Protoplasts of cells from a petunia's
spheroplasts: have their cell wall only partially removed and are gram − leaf
(gram-negative)
more generally, a unit of biology which is composed of a cell's nucleus and
the surrounding protoplasmic materials.

Contents
Enzymes for the preparation of protoplasts
Uses for protoplasts
See also
References
Protoplasts of the moss
Physcomitrella patens

Enzymes for the preparation of

protoplasts
Cell walls are made of a v ariety of poly saccharides. Protoplasts can be made by degrading cell walls with a mixture of
the appropriate poly saccharide-degrading enzy mes:

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 v ery sensitiv e to osmotic stress. This
means cell wall digestion and protoplast storage must be done in an isotonic solution to prev ent rupture of the
plasma membrane.

Uses for protoplasts

Protoplasts can be used to study membrane biology , including the uptake of macromolecules and v iruses . These are
also used in somaclonal v ariation.
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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. [3 ] In the case of plant cells, protoplasts may be
regenerated into whole plants first by growing into a group of plant cells
that dev elops into a callus and then by regeneration of shoots
(caulogenesis) from the callus using plant tissue culture methods. [4 ]
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
Fused protoplast (on left), containing
v ascular plants, protoplasts from mosses, such as Physcomitrella patens,
both chloroplasts (from a leaf cell) as
do not need phy tohormones for regeneration, nor do they form a callus well as a coloured vacuole (from a
during regeneration. Instead, they regenerate directly into the filamentous petal).
protonema, mimicking a germinating moss spore. [5 ]

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 poly ethy lene gly col. [6 ] This technique
may be used to generate somatic hy brids in tissue culture.

Additionally , protoplasts of plants expressing fluorescent proteins in certain cells may be used for Fluorescence
Activ ated Cell Sorting (FACS), where only cells fluorescing a selected wav elength are retained. Among other things,
this technique is used to isolate specific cell ty pes (e.g., guard cells from leav es, pericy cle cells from roots) for
further inv estigations, such as transcriptomics.

See also
L-form bacteria

References
1. Hanstein, J. 1880. Das Protoplasma (https://archive.org/details/DasProtoplasma). Heidelberg.
2. Sharp, L. W. (1921). Introduction To Cytology (https://archive.org/details/introductiontocy032473mbp). New York:
McGraw Hill, p. 24.
3. 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 (https://doi.org/10.1016%2Fj.biotechadv.2004.09.
008). PMID 15694124 (https://www.ncbi.nlm.nih.gov/pubmed/15694124).
4. Thorpe TA (October 2007). "History of plant tissue culture". Mol. Biotechnol. 37 (2): 169–80. doi:10.1007/s12033-007-
0031-3 (https://doi.org/10.1007%2Fs12033-007-0031-3). PMID 17914178 (https://www.ncbi.nlm.nih.gov/pubmed/1791417
8).
5. 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 (http://www.springerlink.com/content/m
747f7xfu9dxyprw/?p=2747894a7a9849a09e60b0e49f51afdc&pi=3Observation). Protoplasma 219, 99–105.
6. 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.

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