Treatment of Bacteria with Plant Root Exudates to Emulate In Vitro Plant-Microbe Interactions

Take a stock culture of Bacillus mycoides, a plant growth-promoting rhizobacterium, and streak an inoculum onto a culture plate.

Incubate the plate to promote bacterial proliferation into colonies.

Inoculate a single colony into a liquid medium and incubate with shaking to promote bacterial growth.

Dilute the culture using the liquid medium to achieve the optimal density for the assay.

Add potato root exudates to the culture to emulate plant-microbe interactions.

Incubate the culture with agitation, facilitating the interaction of the signaling molecules with bacterial receptors.

This interaction induces signaling pathways that activate regulatory proteins, which bind to DNA and alter gene expression.

Transfer the culture into a tube and spin down the bacterial cells into a pellet.

Discard the supernatant and dip the tube into liquid nitrogen to flash-freeze the cells.

Store the cells to analyze plant root exudate-induced changes in bacterial gene expression.

To grow the bacteria, streak a B. mycoides strain from a minus 80 degree Celsius glycerol stock onto an LB agar plate, and incubate the plate at 30 degrees Celsius overnight. Then inoculate the LB liquid medium with a single colony fro