Phenazines are involved in biocontrol of Pythium myriotylum on cocoyam by Pseudomonas aeruginosa PNA1

Root rot of cocoyam (Xanthosoma sagittifolium) caused by Pythium myriotylum is the most devastating disease of this important tropical tuber crop with yield reductions of up to 90%. Bioassays were conducted in vitro and in sterile volcanic soil artificially infested with Pythium myriotylum, isolate CRPm, to test whether Pseudomonas aeruginosa PNA1 can control the cocoyam root rot disease. P. aeruginosa PNA1 (wild type) produces phenazine-1-carboxylic acid and phenazine-1-carboxamide (oxychlororaphin), while its tryptophan auxotrophic mutant FM13 is phenazine negative and secretes anthranilate in vitro. PNA1 and FM13 have previously been shown to control Pythium debaryanum and Pythium splendens on lettuce and bean. PNA1 and FM13 significantly inhibited growth of P. myriotylum in dual cultures, while their supernatants highly reduced mycelial dry weight in potato dextrose broth. However, in the presence of tissue culture derived cocoyam plantlets, only strain PNA1 strongly reduced root rot disease severity. Soil experiments involving strain PNA1 in comparison to phenazine-deficient mutants suggested that the biocontrol activity of PNA1 against P. myriotylum may involve phenazines. Phenazine involvement was further strengthened by the fact that FM13 fed with exogenous tryptophan (so that phenazine production is restored) significantly reduced disease severity on cocoyam. The efficiency of PNA1 to control P. myriotylum on cocoyam was significantly improved when the strain and the pathogen were allowed to interact for 24 h prior to transplanting cocoyam plantlets, while doubling the inoculum density of the pathogen negatively affected its efficiency.