Reference Work Entry

Handbook of Hydrocarbon and Lipid Microbiology

pp 3461-3470

Combined Use of Fluorescent Reporters and Flow Cytometry for Simultaneous Monitoring of Bacterial Growth and Gene Expression on Plant Roots

  • L. RochatAffiliated withDepartment of Fundamental Microbiology, University of Lausanne
  • , M. Péchy-TarrAffiliated withDepartment of Fundamental Microbiology, University of Lausanne
  • , M. MaurhoferAffiliated withInstitute of Integrative Biology, Plant Pathology, Swiss Federal Institute of Technology (ETH)
  • , C. Keel*Affiliated withDepartment of Fundamental Microbiology, University of Lausanne

Abstract:

A protocol of a recently developed assay is presented in which FACS-based flow cytometry is used in combination with autofluorescent reporter constructs to simultaneously monitor and quantify bacterial growth (root colonization levels) and single cell activity (expression of a plant-beneficial trait) on roots of hydroponically grown plants. The experimental approach is illustrated with an example of a reporter of antifungal gene expression in a plant-beneficial strain of Pseudomonas fluorescens. It can be adapted to the study of other bacteria-root interactions.