Microbial biosensors designed with the luxCDABEG
reporter genes (that code for light- emitting
proteins) fused to a promoter of interest offer the opportunity for continuous
measurements of environmental conditions from the soil. We use the common soil bacterium Pseudomonas
putida KT2440 as host to two different plasmids we
developed to report on water potential and microbial growth in the rhizosphere.
The first plasmid pZKH2 contains a fusion between the strong constituitive
promoter npt and the lux genes that reports on growth of the bacteria. The second construct, pPHCA1, utilizes a proU-luxCDABEG fusion to track changes in water potential around
the bacterium. After testing
biosensor response in liquid media, we used both biosensors in microcosms filled
with non-sterile soil in which Zea mays L. was
growing. Using a very sensitive
CCD-camera to view light production through the glass sides of the microcosm,
we detected temporal and spatial patterns in carbon availability. Biosensors
demonstrated pulses of bacteria growth near the root tips likely associated
with the release of carbon. The biosensors were able
to continuously report from around roots for as long as six days at a
time with the ability to follow bacterial activity in real time around growing
roots.