Gene Transfer Methods /Techniques in Plants

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It is a technique to efficiently and stably introduce foreign genes (Gene of interest) into the genome of target cells or transferring a gene into a non native organism and the process is called Transgenesis. There are two methods of gene transfer in plants.
1. Direct or Vector-less Gene Transfer.
a. Physical
i. Particle gun
ii. Electroporation
iii. Microinjection
b. Chemical Method
i. Liposome
ii. PEG
2. Indirect or Vector-Mediated Gene Transfer
a. A.tumefaciens (produces a gall)
b. A. rhizogenes (produces excessive root hairs)
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Particle gun
The foreign DNA containing gene of interest is coated onto beads/surface of either Au or tungsten (1-um) called as micro carrier.
The coated beads are then attached to the end of the plastic bullet (Macro carrier) and loaded into the firing chamber of the gene gun and then bombarded on target cells by particle gun.
This method avoids the need of protoplast and is better in efficiency. This technique can be used for any plant cells, root section, embryos, seeds and pollen or even on intact plant in the experimental filed.
Though some cells get destroyed but on the other side many cell receive foreign DNA and thus can produce large no of transforming plant cell.
Biolistics technique has been used successfully to transform soyabean, cotton, spruce, sugarcane, papaya, sunflower, rice, maize, wheat, tobacco etc
Electroporation
Introduction of DNA into plant cell by exposing them for very brief periods into high voltage electric pulses, which induce transient pores in the plasma membrane, is called electroporation.
If plant cell (Protoplast) is placed in medium containing GoI then the DNA enter into plant cell via pores and finally incorporated into plant genome.
Advantage: Some plants are PEG sensitive so electroporation may be of choice, method is fast, less costly and high percentage of stable transformants can be produced.
It has been used to produce stably transformed cell lines and plants of several species like tobacco, maize, rice, wheat, etc.

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