Serial Dilution-Based Growth Curves and Growth Curve Synchronization for High-Resolution Time Series of Bacterial Biofilm Growth

Abstract

The ability to form stable surface-attached communities called biofilms is of paramount importance to both beneficial and harmful interactions between microbes and microbial, plant or animal partners. Assessment of biofilm formation ability is often performed by growing the organisms in microtiter plate wells and staining the well-attached material, a method whose use for time-course analysis is limited by its destructive nature. Here we combine a serial dilution-based biofilm growth curve method with online monitoring of planktonic growth and a serially diluted growth curve synchronization algorithm to reconstruct the time-course of planktonic and biofilm growth. As demonstrated here with the rhizosphere bacterium Pseudomonas putida, the method allows accurate determination of the growth rate and doubling time, a robust depiction of the biofilm formation and dispersal dynamics and assessment of the biofilm development defects in mutant strains.