Polymorphism of the polyketide synthase gene phlD in biocontrol fluorescent pseudomonads producing 2,4-diacetylphloroglucinol and comparison of PhlD with plant polyketide synthases

Many biocontrol fluorescent pseudomonads can protect plants from soilborne fungal pathogens through production of the antifungal secondary metabolite 2,4-diacetylphloroglucinol (Phl), One of the phl biosynthetic genes, phlD, encodes a polyketide synthase similar to plant chalcone syntheses. Here, re striction analysis of phlD from 39 Phl(+) biocontrol fluorescent pseudomona ds yielded seven different banding patterns. The gene was sequenced in seve n strains, representing the different restriction patterns. Cluster analysi s of phlD restriction data or phlD sequences indicated that phlD polymorphi sm was high, and two main clusters were obtained when predicted PhlD sequen ces were compared. When the seven PhlD sequences were studied with those of other procaryotic polyketide syntheses (gram-positive bacteria) and plant chalcone syntheses, however, Phl+ pseudomonads, grampositive bacteria, and plants clustered separately. Yet, sequence analysis of active site regions for PhlD and plant chalcone synthases revealed that PhlD can be considered a member of the chalcone synthase family, which may be interpreted as conve rgent evolution of key enzymes involved in secondary metabolism. For the 39 PhlD pseudomonads, a relationship was found among phlD restriction pattern s, phylogenetic groups defined by 16S rDNA restriction analysis (confirmed by 16S rDNA sequencing), and production levels of Phl in vitro.