Development of genetic tools in environmental Pseudomonas species

Abstract

Plant growth-promoting rhizobacteria possess many beneficial traits that make them desirable in agricultural applications. Several Pseudomonas spp. have developed a close association with plants and perform nutrient exchange as well as protect the plant from pathogens. Limited research has been done in genetically manipulating these organisms, which could further enhance the agricultural utility of these strains, providing a far less destructive alternative than the chemical fertilizers and pesticides used today. In this work I aimed to build a genetics system in several species of environmental Pseudomonas as the basis for future manipulation and research within these strains. I first investigated growth dynamics and transformation strategies to later introduce phage-derived recombinases for directed chromosomal mutation of the rpoB gene while providing direct comparison of these criteria between different strains of Pseudomonas. I also highlight some of the unique behavior within these Pseudomonas spp. that further emphasizes the individuality between strains and difficulty in creating a “one-size fits all” protocol, while identifying key components to consider when adapting an environmental bacterium to life in the lab.