An antagonistic rhizoplane bacterium Pseudomonas sp. strain EC-S101 physiologically stresses a spinach root rot pathogen Aphanomyces cochlioides

Abstract

We observed that an antagonistic rhizoplane bacterium Pseudomonas sp. strain EC-S101 induces excessive lateral and apical branching in the hyphae of a root rot phytopathogen Aphanomyces cochlioides AC-5 resulting in radial growth inhibition of hyphae in a dual culture assay. Confocal laser scanning microscopic observations using fluorescent stains indicated an increased quantity of nuclei and lipid bodies in the affected hyphae during the early stage (less affected hyphae) at day 3 of interaction. At a more advanced stage (severely affected hyphae) at day 3, nuclei became smaller and round-shaped compared with the oval shape in AC-5 control hyphae. After 7 days, nuclei disintegrated, and the nuclear materials were released into the disorganized cytoplasm. With transmission electron microscopy at 5 days of interaction, we found that the cell walls of AC-5 hyphae were considerably thicker than those of the control. Enlarged vacuoles, lipid bodies sunk into vacuoles, and vacuoles filled with electron-dense material, followed by an invagination of the AC-5 hyphal cell wall, were commonly observed. Nonmembranous electron-transparent inclusion bodies irregular in size were often distributed in the affected hyphae. By integrating our observations, we conclude that antagonistic effects evoked by strain EC-S101 resulted in the death of AC-5 hyphae, which might contribute to the suppression of A. cochlioides AC-5-linked damping-off disease in its host plants.