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We have previously demonstrated that inoculation of tomato plants with 2,4-diacetylphloroglucinol (DAPG)- and hydrogen cyanide (HCN)-producingPseudomonas brassicacearum LBUM300 could significantly reduce bacterial canker symptoms caused byClavibacter michiganensis subsp.michiganensis . In this study, in order to better characterize the population dynamics of LBUM300 in the rhizosphere of tomato plants, we characterized the role played by DAPG and HCN production by LBUM300 on rhizosphere colonization of healthy andC. michiganensis subsp.michiganensis -infected tomato plants. The impact ofC. michiganensis subsp.michiganensis presence on the expression of DAPG and HCN biosynthetic genes in the rhizosphere was also examined.In planta assays were performed using combinations ofC. michiganensis subsp.michiganensis and wild-type LBUM300 or DAPG (LBUM300ΔphlD ) or HCN (LBUM300ΔhcnC ) isogenic mutant strains. Populations of LBUM300 andphlD andhcnC gene expression levels were quantified in rhizosphere soil at several time points up to 264 h postinoculation using culture-independent quantitative PCR (qPCR) and reverse transcriptase quantitative PCR (RT-qPCR) TaqMan assays, respectively. The presence ofC. michiganensis subsp.michiganensis significantly increased rhizospheric populations of LBUM300. InC. michiganensis subsp.michiganensis -infected tomato rhizospheres, the populations of wild-type LBUM300 and strain LBUM300ΔhcnC , both producing DAPG, were significantly higher than the population of strain LBUM300ΔphlD . A significant upregulation ofphlD expression was observed in the presence ofC. michiganensis subsp.michiganensis , whilehcnC expression was only slightly increased in the mutant strain LBUM300ΔphlD whenC. michiganensis subsp.michiganensis was present. Additionally, biofilm production was found to be significantly reduced in strain LBUM300ΔphlD compared to the wild-type and LBUM300ΔhcnC strains.IMPORTANCE The results of this study suggest thatC. michiganensis subsp.michiganensis infection of tomato plants contributes to increasing rhizospheric populations of LBUM300, a biocontrol agent, as well as the overexpression of the DAPG biosynthetic operon in this bacterium. The increasing rhizospheric populations of LBUM300 represent one of the key factors in controllingC. michiganensis subsp.michiganensis in tomato plants, as DAPG-producing bacteria have shown the ability to decrease bacterial canker symptoms in tomato plants.

applied and environmental microbiology

Interaction between 2,4-Diacetylphloroglucinol- and Hydrogen Cyanide-Producing Pseudomonas brassicacearum LBUM300 and Clavibacter michiganensis subsp. michiganensis in the Tomato Rhizosphere