Exopolysaccharide Production and Precipitation Method as a Tool to Study Virulence Factors

Acidovorax avenae subsp. avenae (Aaa) is the causal agent of red stripe in sugarcane, a disease characterized by two forms: leaf stripe and top rot. Despite the importance of this disease, little is known about Aaa virulence factors (VFs) and their function in the infection process. Among the different array of VFs exerted by phytopathogenic bacteria, exopolysaccharides (EPSs) often confer a survival advantage by protecting the cell against abiotic and biotic stresses, including host defensive factors. They are also main components of the extracellular matrix involved in cell-cell recognition, surface adhesion, and biofilm formation. EPS composition and properties have been well studied for some plant pathogenic bacteria; nevertheless, there is no knowledge about Aaa-EPS. In this work, we describe a simple and reliable method for EPS production, precipitation, and quantification based on cold precipitation after ethanol addition, which will allow to study EPS characteristics of different Aaa strains and to evaluate the association among EPS (e.g., amount, composition, viscosity) and Aaa pathogenicity.