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Chromosomal Insertion of Phenazine-1-Carboxylic Acid Biosynthetic Pathway Enhances Efficacy of Damping-off Disease Control by Pseudomonas fluorescens

December 2000 , Volume 13 , Number  12
Pages  1,293 - 1,300

T. M. Timms-Wilson , 1 R. J. Ellis , 1 A. Renwick , 2 D. J. Rhodes , 2 D. V. Mavrodi , 3 D. M. Weller , 3 L. S. Thomashow , 3 and M. J. Bailey 1

1Molecular Microbial Ecology, NERC Institute of Virology and Environmental Microbiology, Mansfield Road, Oxford, OX1 3SR, U.K.; 2Zeneca Agrochemicals, Jealott's Hill Research Station, Bracknell, Berkshire, RG12 6EY, U.K.; 3USDA-ARS, Department of Plant Pathology, Washington State University, Pullman 99164-6430, U.S.A.

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Accepted 12 August 2000.

A disarmed Tn5 vector (pUT∷Ptac-phzABCDEFG) was used to introduce a single copy of the genes responsible for phenazine-1-carboxylic acid (PCA) biosynthesis into the chromosome of a plant-growth-promoting rhizobacterium Pseudomonas fluorescens. The PCA gene cluster was modified for expression under a constitutive Ptac promoter and lacked the phzIR regulators. PCA-producing variants significantly improved the ability of the wild-type P. fluorescens to reduce damping-off disease of pea seedlings caused by Pythium ultimum, even under conditions of heavy soil infestation. Under conditions of oxygen limitation that are typical of the rhizosphere, PCA production per cell in vitro was greater than that recorded in fast-growing, nutrient-rich cultures. Similarly, when the in vitro nutrient supply was limited, P. fluorescens∷phz variants that produced the most PCA effectively competed against P. ultimum by suppressing mycelial development. Soil-based bioassays confirmed that the level of PCA biosynthesis correlated directly with the efficacy of biological control and the persistence of inocula in soil microcosms. They also showed that soil pretreatment with bacteria provides a suitable method for plant protection by reducing infection, effectively decontaminating the soil. These data demonstrate that the insertion of a single chromosomal copy of the genes for a novel antifungal compound, PCA, enhances the ecological fitness of a natural isolate already adapted to the rhizosphere and capable of suppressing fungal disease.

© 2000 The American Phytopathological Society