Transferring genes to plants

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Lecture 4

• Transferring genes to plants

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Monocotyledones are not easy to handle callus
is very difficult to be initiated, and
A.tumefaciens is not pathogenic for them
1. Pericarp sholud be pulled back and the
immature embryos (0.5 - 1.0 mm) are removed.
2. The immature embryos are placed on a callus
induction medium
Transformation is performed by gene gun method
high osmotic media prepare calli for
transfomation
plantsciences.montana.edu/ .../transform1.htm
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DNA with desired gene and antibiotic resistance
is coated onto the surface of gold particles.
Calli are placed in vacuum chamber, Helium
pressure shot DNA into cells
Gene gun
Coating gold particles with DNA
vacuum chamber
Calli remain on the high osmotic media for 20
hours following shooting.
plantsciences.montana.edu/ .../transform1.htm
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Closer look on (gene gun)
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1. DNA- or RNA-coated gold/tungsten particles are
   loaded into the gun and you pull the trigger.
2. A low pressure helium pulse delivers the coated
   gold/tungsten particles into virtually any target
   cell or tissue.
3. The particles carry the DNA ? cells do not have
   to be removed from tissue in order to transform
   the cells
4. As the cells repair their injuries, they
   integrate their DNA into their genome, thus
   allowing for the host cell to transcribe and
   translate the transgene.

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Alternate Methods of Transforming Plants
Particle Bombardment
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After shooting calli are placed on a selective
media containing a herbicide for three weeks.
Then calli are transferred to a media to induce
the production of shoots. After they form small
shoots, they are transferred to DARKER
containers on a root induction media.
plantsciences.montana.edu/ .../transform1.htm
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plantsciences.montana.edu/ .../transform1.htm
The small plantlets are transplanted into soil
and acclimated under high humidity conditions
With current procedures only 10-20 of the plants
are actually transgenic, so they should be
tested on transgene expression
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