root tissue culture system to study winter wheat-rhizobacteria interactions

A tissue culture procedure was developed for growing winter wheat roots. This system was used to study bacterial-root interactions and root colonization by Pseudomonas cepacia strains R55 and R85, Azospirillum brasilense ATCC 29729 and Azotobacter chroococcum ATCC 9043. Axenic root tissue cultures were inoculated with bacteria and incubated at 25 degrees C on a rotary shaker (150 rpm) for up to 3 weeks. At various intervals, root morphology and root hair development, bacterial colonization of root surfaces and nitrogenase activity were determined. As determined by plate counting techniques, bacterial attachment to the roots varied from 7.5 X 10(4) to 3.2 X 10(7) colony-forming units cm-1. Scanning electron microscopy of inoculated roots revealed that some rhizobacteria (e.g., P. cepacia R85) significantly enhanced root hair development, and others (e.g., P. cepacia R55) specifically colonized root exudation sites. Nitrogenase activity of roots inoculated with either A. brasilense ATCC 29729 or A. chroococcum ATCC 9043 was stimulated if the inorganic nitrogen sources in the medium were replaced with 300 microgram glutamine ml-1. We propose that the use of root tissue culture systems will facilitate studies on plant root-microbe interactions.