MGH Plant Growth Facility SOP
Biosafety Guidelines for Handling
Agrobacterium for Plant Transformation
MGH Molecular Biology Department
Jenifer Bush, Greenhouse Manager 617-726-5931
The Agrobacterium strain that we use in the laboratory has been altered from
the form found in nature to substantially reduce its virulence and any
environmental harm. For example, it is not able to form tumors on host plants
because the majority of crown gall tumor-forming related genes have been
deleted from the Ti plasmid.
The ability of A. tumefaciens to insert genes into a plant is what makes it such a
useful tool in the laboratory. However, this trait may still be a risk factor for the
environment.
To prevent any unintentional release of A. tumefaciens strains into the
environment, adhere to these guidelines.
1. Only use disarmed non-tumorigenic Ti plasmid vectors and non-pathogenic
viral vectors in A. tumefaciens for stable or transient plant transformation.
2. The leftover A. tumefaciens cultures must be rendered biologically inactive by
either autoclaving them or mixing them with an equal volume of bleach for 1
hour.
3. Any spilled A. tumefaciens culture should be wiped up, and the surface
cleaned with 70% ethanol or 50% concentration bleach. Towels or pads used to
clean the spill should then be autoclaved.
4. Plants that are treated or infiltrated with A. tumefaciens should be autoclaved
after seed harvest or when the experiment is complete.
5. Consider using a minimal amount of A. tumefaciens by pipetting A.
tumefaciens cultures onto flowers instead of dipping whole plants.
Updated March 2007 JB