A high-throughput method for quantifying growth of phytopathogenic bacteria in Arabidopsis thaliana.
Measuring the growth of pathogenic bacteria in leaves is a mainstay of plant pathology studies. We have made significant improvements to standard methods that will not only increase the throughput but also reduce the space limitations. Additionally, the method described here is as accurate as the standard method. Briefly, we infected leaves by dipping whole seedlings of Arabidopsis into a bacterial solution containing a surfactant. After harvest, the seedlings were then simply shaken in buffer. The resulting bacterial solutions were diluted in microtitre plates and spotted onto agar plates. Colony-forming units were then counted 40 h after plating. Therefore, we have eliminated most of the labour-intensive steps involved in measuring the growth of bacteria in Arabidopsis, and describe a method that could be automated. The assay is sensitive enough to detect small differences between pathogens or ecotypes.
Tornero
P
P
Department of Biology and Curriculum in Genetics, Coker Hall 108, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Dangl
J L
JL
D017360:Arabidopsis / Q000382:microbiology*; D015169:Colony Count, Microbial / Q000379:methods*; D010935:Plant Diseases / Q000382:microbiology*; D011549:Pseudomonas / Q000302:isolation & purification*