1. Plant Genetic Transformation

2. All stable transformation methods consist of three steps:
Delivery of DNA into a single plant cell.
Integration of the DNA into the plant cell genome.
Conversion of the transformed cell into a whole plant.

3. Methods of delivering DNA into plant cells
Biological
Agrobacterium
Other bacteria
Viruses
Physical
Particle bombardment
Electroporation
Silicon carbide whiskers
Carbon nanofibers

5. Agrobacterium-mediated Transformation

6. Biology of the Agrobacterium-plant interaction
The only known natural example of inter-kingdom DNA transfer

7. Infects at root crown or just below the soil line.
Can survive independent of plant host in the soil.
Infects plants through breaks or wounds.
Common disease of woody shrubs, herbaceous plants, dicots.
Galls are spherical wart-like structures similar to tumors.

10. The genus Agrobacterium has a wide host range:
Overall, Agrobacterium can transfer T-DNA to a broad group of plants.
Yet, individual Agrobacterium strains hve a limited host range.
The molecular basis for the strain-specific host range is unknown.
Many monocot plants can be transformed (now), although they do not form crown gall tumors.
Under lab conditions, T-DNA can be transferred to yeast, other fungi, and even animal and human cells.

11. Why is Agrobacterium used for producing transgenic plants?
The T-DNA element is defined by its borders but not the sequences within. So researchers can substitute the T-DNA coding region with any DNA sequence without any effect on its transfer from Agrobacterium into the plant.

14. Ti Plasmid

26. Key steps from natural Agrobacterium to “useful”Agrobacterium
Some vir genes deleted—disarmed
Opines not going to be produced
Deleting tumorogenesis function
Choosing strains that transfer DNA in lab
Clone in genes of interest, antibiotic resistance genes, etc.
Binary system-- two plasmids are better than one Ti plasmid

30. Arabidopsis Floral Dip

31. Biolistic Transformation Gene Gun

35. Agroinfiltration Transient method of expressing transgenes