A Co-Cultivation System to Study the Molecular Responses of Plant Hosts and Interacting Bacteria

Take a tank containing a sterile growth medium with an acidic pH.

Transfer a mesh screen with plant seedlings into the tank.

Seal with a lid and porous tape to maintain sterility while allowing gaseous exchange.

Incubate with shaking to aerate the medium.

Inspect the medium to check for the absence of cloudiness, indicating no bacterial contamination.

Spot a sample onto a culture plate and incubate it to observe the lack of bacterial growth, confirming the absence of contaminating bacteria.

Inoculate with Agrobacterium, a plant pathogen, and incubate.

Plant root-derived signaling molecules and the acidic pH activate a bacterial membrane-bound sensor protein.

The activation induces gene expression, producing proteins for bacterial attachment to root surfaces.

Following attachment, Agrobacterium delivers a DNA fragment, known as transfer DNA (T-DNA), into the plant cells via a secretion system.

Upon integration into the plant genome, the T-DNA synthesizes signaling molecules that enhance bacterial colonization.

In a sterilized flow hood, pour 18 milliliters of autoclaved liquid MS medium into sterile cylindrical glass tanks with sterile lids.

Next, use sterile forceps to gently transfer each mes