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Protoplast Production Methods in Plants

This document discusses methods for producing protoplasts from plant tissue. It describes protoplasts as naked plant cells without cell walls but with other cellular components. Three methods for producing protoplasts are discussed: 1) the mechanical method, which is crude and yields few protoplasts; 2) the enzymatic method, which uses enzyme mixtures to dissolve cell walls and yields more viable protoplasts; and 3) the two-step and one-step enzymatic methods, where the two-step method uses pectinase followed by cellulose and the one-step uses both simultaneously.

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21 views5 pages

Protoplast Production Methods in Plants

This document discusses methods for producing protoplasts from plant tissue. It describes protoplasts as naked plant cells without cell walls but with other cellular components. Three methods for producing protoplasts are discussed: 1) the mechanical method, which is crude and yields few protoplasts; 2) the enzymatic method, which uses enzyme mixtures to dissolve cell walls and yields more viable protoplasts; and 3) the two-step and one-step enzymatic methods, where the two-step method uses pectinase followed by cellulose and the one-step uses both simultaneously.

Uploaded by

Andrew
Copyright
© All Rights Reserved
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NAME: TANYARADZWA O CHABHONGORA

REG NUMBER: C19138019S


PROGRAMME: BSCST
COURSE CODE: CUBT 207
QUESTION
Comment Assignment is the same as tht of
CHIKATI VIMBISO

Discuss any 3 methods used to produce protoplast


from plant tissue.
Protoplasts are naked plant cells without the cell
wall, but they possess plasma membrane and all
other cellular [Link] of different
species can be fused to generate a hybrid and this
process is referred to as somatic
[Link] used to produce
protoplast from plant tissue include mechanical
method and the enzymatic [Link]
isolation by mechanical method is crude and
tedious [Link] results in the isolation of a
very small number of [Link] involves
plasmolysis of a suitable tissue such as onion bulb
epidermis, by immersion in 1,0 molar
sucrose,followed by slicing the tissue and finally
deplasmolysis in 0,4M [Link]
deplasmolysis a few surviving proplasts from
higher plant tissues but are effective with certain
filamentous algae. Protoplasts have been isolated
from higher plants, mosses(Binding 1966),
liverworts(Schieder 1975), fungi( Ferency et
al.1957),blue-green algae(Crespi et
al1976).Spheroplasts- protoplasts that retain part
of their lipo-polyssaccharide wall components
have been prepared from the green alga
chlorella(Brown Aach 1975).The enzymatic
method avoid the shortcomings of mechanical
methods and can yield large numbers of
protoplasts( Cocking 1960). It is rhe application of
crude fungal enzyme mixtures to exposed plant
tissue caused dissolution of cell walls followed by
the emergence of naked [Link]
advantages of enzymatic method include good
yield of viable cells and minimal or no damage to
the [Link] can be isolated from a
wide variety of tissues and organs that include
leaves,roots,shoot apices,fruits
[Link] shoots are most frequently
[Link] plant cell wall is degradedby the
enzymescellulase, hemicellulose and pectinase ,
the enzymes are usually used at a pH 4,5 to6,0,
temperature 25-30 with a wide variation in
incubation period that range from half an hour to
20 [Link] enzymatic isolation of protoplasts
can be carried out by two approaches ,the two
step or sequential method, the tissue is first
treated with pectinase to separate cells by
degrading middle [Link] free cells are then
exposed to cellulose to release protoplasts.
Pectinase breaks up the cell aggregates into
individual cells while cellulose removes the cell
wall proper and the one step or simultsneous
[Link] is the preferred method for
protoplast [Link] involves the simultaneous
use of both the enzymes and cellulose.
REFERENCE LIST

[Link] R, Cocking EC, Thompson JA (1986)


Efficient plant regeneration from rice protoplasts
through somatic embryogenesis. Biotechnology
4:1087–1090
CrossRefGoogle Scholar
[Link] MA, Stone BA (1975) A new substrate
for investigating the specificity of β-glucan
hydrolases. FEBS Lett 52:202–207
PubMedCrossRefGoogle Scholar
[Link] YPS (1977) Protoplasts isolation, culture
and somatic hybridization. In: Reinert J, Bajaj YPS
(eds) Applied and fundamental aspects of plant
cell, tissue, and organ culture. Springer, Berlin
Heidelberg New York, pp 467

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