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PEG Method for Gene Transfer in Plants

Polyethylene glycol (PEG) is used to induce DNA uptake into plant protoplasts for gene delivery. PEG treatment permeabilizes protoplast membranes, allowing plasmid DNA to enter the cell and integrate into the plastome. Specifically, PEG compacts DNA and associates it with cell membranes, facilitating DNA transfer into protoplasts. While this PEG method allows large-scale transformation of protoplasts, regeneration of transformed plants remains challenging and random DNA integration can cause undesirable traits.

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Etti Singh
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2K views9 pages

PEG Method for Gene Transfer in Plants

Polyethylene glycol (PEG) is used to induce DNA uptake into plant protoplasts for gene delivery. PEG treatment permeabilizes protoplast membranes, allowing plasmid DNA to enter the cell and integrate into the plastome. Specifically, PEG compacts DNA and associates it with cell membranes, facilitating DNA transfer into protoplasts. While this PEG method allows large-scale transformation of protoplasts, regeneration of transformed plants remains challenging and random DNA integration can cause undesirable traits.

Uploaded by

Etti Singh
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd

PEG METHOD OF

GENE DELIVERY
Presented by : Etti Singh
M.Sc. Biochemistry, 4th Semester
Plant Biotechnology
S.O.S. in Life Sciences, PTRSU, Raipur(C.G.)
WHAT IS PEG?

Polyethylene glycol is a polyether compound derived from petroleum.


PEG is also known as polyethylene oxide or polyoxyethylene,
depending on its molecular weight. It can be used to induce DNA
uptake into plant protoplasts.

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HOW DOES PEG METHOD OF
GENE TRANSFER WORK?
•  PEG-mediated transformation of plastids requires enzymatically removing the cell wall to obtain
protoplasts, then exposing the protoplasts to purified DNA in the presence of PEG.
•  The protoplasts first shrink in the presence of PEG, then lyse due to disintegration of the cell
membrane. Removing PEG before the membrane is irreversibly damaged reverses the process.
•  Treatment of freshly isolated protoplasts with PEG allows permeabilization of the plasma membrane
and facilitates uptake of DNA
• Plasmid DNA passes the plastid membranes and reaches the stroma where it integrates into the
plastome as during biolistic transformation.
•  A relatively small number of species have been transformed using this approach, mainly because it
requires efficient isolation, culture and regeneration of protoplasts, a tedious and technically demanding
in vitro technology.

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WHAT PEG DOES TO FACILITATE
THE GENE TRANSFER?

• PEG changes the DNA conformation, thus it compacts the DNA and also allows it to
associate with the cell membrane.

• PEG is also used as an in vitro crowding agent to mimic highly crowded cellular
conditions.

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ADVANTAGES OF PEG METHOD OF GENE TRANSFER

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 A large number of protoplasts can be simultaneously transformed.


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 This can be successfully used for a wide range of plant species with adequate
modifications.

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DISADVANTAGES OF PEG METHOD OF GENE TRANSFER

 The DNA is susceptible for degradation and rearrangement.


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 Random integration of foreign DNA into genome may result in


undesirable traits.
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 Regeneration of plants from transformed protoplasts is a difficult task.


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REFERENCES

• Bilang,
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R., Klöti, A., Schrott, M. and Potrykus, I., 1994. PEG-mediated direct gene
transfer and electroporation. In Plant molecular biology manual (pp. 1-16). Springer,
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Dordrecht.
• Craig, W., Gargano, D., Scotti, N., Nguyen, T.T., Lao, N.T., Kavanagh, T.A., Dix, P.J.
and
Place Cardi, T., 2005. Direct gene transfer in potato: a comparison of particle

bombardment of leaf explants and PEG-mediated transformation of


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protoplasts. Plant cell reports, 24(10), pp.603-611.


• Mathur, J. and Koncz, C., 1998. PEG-mediated protoplast transformation with naked
DNA.
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In Arabidopsis Protocols (pp. 267-276). Humana Press.
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THANK
YOU
ETTI SINGH
M.Sc. Biochemistry 4th Semester

Plant Biotechnology

S.O.S, in Life Sciences, PTRSU,


Raipur (C.G.)

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