Presenting Author Iowa State University, Ames Labarotory
Rhizosphere, the narrow region surrounding plant roots directly influenced by root exudates and the root associated microbiome, plays an important role in plant productivity and the rhizosphere is well known for stimulating microbial metabolic activities. How microbial communities interact to form stable, metabolically interconnected functional communities is an area of intense interest. The question remains on how microbially produced secreted molecules that function as intercellular communication signals shape the structure and function of microbial communities. Our goal is to detect and quantify signal molecules produced by microbes or plants in the root-soil environment, spatially and temporarily. As a proof-of-concept, we are imaging diffusible extracellular microbial metabolites involved in a bacteria-bacteria communication process called quorum-sensing. Quorum-sensing relies on the accumulation of high concentrations of signal molecules in the environment to control bacterial gene expression, influencing rhizosphere colonization and plant health. Local concentration of quorum sensing molecules are determined using aptamer based sensors. Aptamers that can specifically bind the desired signal molecules are selected through SELEX and immobilized on the surface of nano-porous membranes. Binding of the signal molecules and aptamer covered surface results in changes in surface charge distribution and steric hinderance and thus modifying the transmembrane ionic transport. Changes in transmembrane impedance can be measured through electrochemical impedance spectroscopy methods to monitor local concentrations of signal molecules. Sensor responses were determined for different concentrations of signal molecules and results showed detection of C4-HSL in a soil-mimic solution with KD of 10 nM. We demonstrate that the quorum-sensing signal molecule C4-homoserine lactone and (C4-HSL) can rapidly diffuse in a soil-mimic gel, providing evidence for our use of a soil-mimic for further aptasensor development and validation. The sensors were then inserted into soil mimic gel for monitoring C4-HSL diffusion and the resulted impedance changes were used to determine the C4-HSL concentration at different positions in the gel. Measurements of local C4-HSL concentrations variation and numerical solution of diffusion equation were used to create 4D images of C4-HSL molecule diffusion in the soil-mimic gel.
This research is supported by the U.S. Department of Energy, Office of Science, Biological and Environmental Research (BER) through the Ames Laboratory. The Ames Laboratory is operated for the U.S. Department of Energy by Iowa State University under Contract No. DE-AC02-07CH11358
