Interactions Between Strains of 2,4-Diacetylphloroglucinol-Producing Pseudomonas fluorescens in the Rhizosphere of Wheat.

ABSTRACT Strains of fluorescent Pseudomonas spp. that produce the antibiotic 2,4-diacetylphoroglucinol (2,4-DAPG) are among the most effective rhizobacteria controlling diseases caused by soilborne pathogens. The genotypic diversity that exists among 2,4-DAPG producers can be exploited to improve rhizosphere competence and biocontrol activity. Knowing that D-genotype 2,4-DAPG-producing strains are enriched in some take-all decline soils and that P. fluorescens Q8r1-96, a representative D-genotype strain, as defined by whole-cell repetitive sequence-based polymerase chain reaction (rep-PCR) with the BOXA1R primer, is a superior colonizer of wheat roots, we analyzed whether the exceptional rhizosphere competence of strain Q8r1-96 on wheat is characteristic of other D-genotype isolates. The rhizosphere population densities of four D-genotype strains and a K-genotype strain introduced individually into the soil were significantly greater than the densities of four strains belonging to other genotypes (A, B, and L) and remained above log 6.8 CFU/g of root over a 30-week cycling experiment in which wheat was grown for 10 successive cycles of 3 weeks each. We also explored the competitive interactions between strains of different genotypes inhabiting the same soil or rhizosphere when coinoculated into the soil. Strain Q8r1-96 became dominant in the rhizosphere and in nonrhizosphere soil during a 15-week cycling experiment when mixed in a 1:1 ratio with either strain Pf-5 (A genotype), Q2-87 (B genotype), or 1M1-96 (L genotype). Furthermore, the use of the de Wit replacement series demonstrated a competitive disadvantage for strain Q2-87 or strong antagonism by strain Q8r1-96 against Q2-87 in the wheat rhizosphere. Amplified rDNA restriction analysis and sequence analysis of 16S rDNA showed that species of Arthrobacter, Chryseobacterium, Flavobacterium, Massilia, Microbacterium, and Ralstonia also were enriched in culturable populations from the rhizosphere of wheat at the end of a 30-week cycling experiment in the presence of 2,4-DAPG producers. Identifying the interactions among 2,4-DAPG producers and with other indigenous bacteria in the wheat rhizosphere will help to elucidate the variability in biocontrol efficacy of introduced 2,4-DAPG producers and fluctuations in the robustness of take-all suppressive soils.

[1]  M. Zala,et al.  Biocontrol of soil-borne fungal plant diseases by 2,4-diacetylphloroglucinol-producing fluorescent pseudomonads with different restriction profiles of amplified 16S rDNA , 1998, European Journal of Plant Pathology.

[2]  J. T. de Souza,et al.  Frequency, Diversity, and Activity of 2,4-Diacetylphloroglucinol-Producing Fluorescent Pseudomonas spp. in Dutch Take-all Decline Soils. , 2003, Phytopathology.

[3]  M. Mazzola,et al.  Wheat Genotype-Specific Induction of Soil Microbial Communities Suppressive to Disease Incited by Rhizoctonia solani Anastomosis Group (AG)-5 and AG-8. , 2002, Phytopathology.

[4]  J. Vanderleyden,et al.  Flagella-driven chemotaxis towards exudate components is an important trait for tomato root colonization by Pseudomonas fluorescens. , 2002, Molecular plant-microbe interactions : MPMI.

[5]  B. Landa,et al.  Identification of Differences in Genome Content among phlD-Positive Pseudomonas fluorescens Strains by Using PCR-Based Subtractive Hybridization , 2002, Applied and Environmental Microbiology.

[6]  D. Wood,et al.  Survival of GacS/GacA Mutants of the Biological Control Bacterium Pseudomonas aureofaciens 30-84 in the Wheat Rhizosphere , 2002, Applied and Environmental Microbiology.

[7]  B. Landa,et al.  Differential Ability of Genotypes of 2,4-Diacetylphloroglucinol-Producing Pseudomonas fluorescens Strains To Colonize the Roots of Pea Plants , 2002, Applied and Environmental Microbiology.

[8]  F. O'Gara,et al.  Phenotypic Selection and Phase Variation Occur during Alfalfa Root Colonization by Pseudomonas fluorescens F113 , 2002, Journal of bacteriology.

[9]  B. Landa,et al.  Comparison of Three Methods for Monitoring Populations of Different Genotypes of 2,4-Diacetylphloroglucinol-Producing Pseudomonas fluorescens in the Rhizosphere. , 2002, Phytopathology.

[10]  B. M. Gardener,et al.  Microbial populations responsible for specific soil suppressiveness to plant pathogens. , 2002, Annual review of phytopathology.

[11]  C. Keel,et al.  The Sigma Factor AlgU (AlgT) Controls Exopolysaccharide Production and Tolerance towards Desiccation and Osmotic Stress in the Biocontrol Agent Pseudomonas fluorescensCHA0 , 2001, Applied and Environmental Microbiology.

[12]  M. Zala,et al.  Cosmopolitan distribution of phlD-containing dicotyledonous crop-associated biocontrol pseudomonads of worldwide origin , 2001 .

[13]  D. Weller,et al.  Changes in Populations of Rhizosphere Bacteria Associated with Take-All Disease of Wheat , 2001, Applied and Environmental Microbiology.

[14]  F. O'Gara,et al.  Impact of 2,4-Diacetylphloroglucinol-Producing Biocontrol StrainPseudomonas fluorescens F113 on Intraspecific Diversity of Resident Culturable Fluorescent Pseudomonads Associated with the Roots of Field-Grown Sugar Beet Seedlings , 2001, Applied and Environmental Microbiology.

[15]  D. Weller,et al.  Exploiting Genotypic Diversity of 2,4-Diacetylphloroglucinol-Producing Pseudomonas spp.: Characterization of Superior Root-Colonizing P. fluorescensStrain Q8r1-96 , 2001, Applied and Environmental Microbiology.

[16]  B. Lugtenberg,et al.  Molecular determinants of rhizosphere colonization by Pseudomonas. , 2001, Annual review of phytopathology.

[17]  B L Maidak,et al.  The RDP-II (Ribosomal Database Project) , 2001, Nucleic Acids Res..

[18]  B. M. Gardener,et al.  A rapid polymerase chain reaction-based assay characterizing rhizosphere populations of 2,4-diacetylphloroglucinol-producing bacteria. , 2001, Phytopathology.

[19]  L. Thomashow,et al.  Genetic Diversity of phlD from 2,4-Diacetylphloroglucinol-Producing Fluorescent Pseudomonas spp. , 2001, Phytopathology.

[20]  D. Crawford,et al.  Use of Randomly Amplified Polymorphic DNA as a Means of Developing Genus- and Strain-Specific Streptomyces DNA Probes , 2000, Applied and Environmental Microbiology.

[21]  S. Kalloger,et al.  Genotypic and Phenotypic Diversity of phlD-ContainingPseudomonas Strains Isolated from the Rhizosphere of Wheat , 2000, Applied and Environmental Microbiology.

[22]  R. Fani,et al.  Frequency and Biodiversity of 2,4-Diacetylphloroglucinol-Producing Bacteria Isolated from the Maize Rhizosphere at Different Stages of Plant Growth , 2000, Applied and Environmental Microbiology.

[23]  J. Buyer,et al.  Importance of pfkA for Rapid Growth ofEnterobacter cloacae during Colonization of Crop Seeds , 2000, Applied and Environmental Microbiology.

[24]  Kevin P. Smith,et al.  HOST VARIATION FOR INTERACTIONS WITH BENEFICIAL PLANT-ASSOCIATED MICROBES. , 1999, Annual review of phytopathology.

[25]  L. Thomashow,et al.  Identification and Characterization of a Gene Cluster for Synthesis of the Polyketide Antibiotic 2,4-Diacetylphloroglucinol from Pseudomonas fluorescens Q2-87 , 1999, Journal of bacteriology.

[26]  Kevin P. Smith,et al.  Genetic basis in plants for interactions with disease-suppressive bacteria. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[27]  S. Kalloger,et al.  Genotypic and phenotypic diversity of PhlD-containing Pseudomonas , 1999 .

[28]  J. V. van Elsas,et al.  Comparison of Paenibacillus azotofixans Strains Isolated from Rhizoplane, Rhizosphere, and Non-Root-Associated Soil from Maize Planted in Two Different Brazilian Soils , 1998, Applied and Environmental Microbiology.

[29]  D. Glandorf,et al.  Role of the O-antigen of lipopolysaccharide, and possible roles of growth rate and of NADH:ubiquinone oxidoreductase (nuo) in competitive tomato root-tip colonization by Pseudomonas fluorescens WCS365. , 1998, Molecular plant-microbe interactions : MPMI.

[30]  K. Toyota,et al.  Effects of bacterial colonization of tomato roots on subsequent colonization by Pseudomonas fluorescens MelRC2Rif , 1998 .

[31]  D. Weller,et al.  Natural plant protection by 2,4-diacetylphloroglucinol-producing Pseudomonas spp. in take-all decline soils , 1998 .

[32]  B. Duffy,et al.  Zinc Improves Biocontrol of Fusarium Crown and Root Rot of Tomato by Pseudomonas fluorescens and Represses the Production of Pathogen Metabolites Inhibitory to Bacterial Antibiotic Biosynthesis. , 1997, Phytopathology.

[33]  F. O'Gara,et al.  Ecological interaction of a biocontrol Pseudomonas fluorescens strain producing 2,4-diacetylphloroglucinol with the soft rot potato pathogen Erwinia carotovora subsp. atroseptica , 1997 .

[34]  F. O'Gara,et al.  Role of 2,4-Diacetylphloroglucinol in the Interactions of the Biocontrol Pseudomonad Strain F113 with the Potato Cyst Nematode Globodera rostochiensis , 1997, Applied and environmental microbiology.

[35]  J. M. Wood,et al.  Osmoadaptation by rhizosphere bacteria. , 1996, Annual review of microbiology.

[36]  C. Keel,et al.  Conservation of the 2,4-diacetylphloroglucinol biosynthesis locus among fluorescent Pseudomonas strains from diverse geographic locations , 1996, Applied and environmental microbiology.

[37]  F. O'Gara,et al.  Mutational Disruption of the Biosynthesis Genes Coding for the Antifungal Metabolite 2,4-Diacetylphloroglucinol Does Not Influence the Ecological Fitness of Pseudomonas fluorescens F113 in the Rhizosphere of Sugarbeets , 1995, Applied and environmental microbiology.

[38]  A. Jagendorf,et al.  Molecular mechanisms of defense by rhizobacteria against root disease. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[39]  P. Lemanceau,et al.  Effect of Two Plant Species, Flax (Linum usitatissinum L.) and Tomato (Lycopersicon esculentum Mill.), on the Diversity of Soilborne Populations of Fluorescent Pseudomonads , 1995, Applied and environmental microbiology.

[40]  S. Lindow,et al.  Ecological Similarity and Coexistence of Epiphytic Ice-Nucleating (Ice+) Pseudomonas syringae Strains and a Non-Ice-Nucleating (Ice-) Biological Control Agent , 1994, Applied and environmental microbiology.

[41]  J. G. Hancock,et al.  Growth patterns and metabolic activity of pseudomonads in sugar beet spermospheres: relationship to pericarp colonization by Pythium ultimum , 1994 .

[42]  P. Bakker,et al.  Siderophore receptor PupA as a marker to monitor wild-type Pseudomonas putida WCS358 in natural environments , 1994, Applied and environmental microbiology.

[43]  D. Weller,et al.  Use of mixtures of fluorescent pseudomonads to suppress take-all and improve the growth of wheat , 1994 .

[44]  S. Lindow,et al.  Invasion and Exclusion among Coexisting Pseudomonas syringae Strains on Leaves , 1993, Applied and environmental microbiology.

[45]  F. O'Gara,et al.  Traits of fluorescent Pseudomonas spp. involved in suppression of plant root pathogens. , 1992, Microbiological reviews.

[46]  M. Mazzola,et al.  Contribution of phenazine antibiotic biosynthesis to the ecological competence of fluorescent pseudomonads in soil habitats , 1992, Applied and environmental microbiology.

[47]  T. Heulin,et al.  Genetic and Phenotypic Diversity of Bacillus polymyxa in Soil and in the Wheat Rhizosphere , 1992, Applied and environmental microbiology.

[48]  C. Keel,et al.  Suppression of root diseases by Pseudomonas fluorescens CHA0 - importance of the bacterial seconday metabolite 2,4-diacetylphloroglucinol , 1992 .

[49]  M. Mazzola,et al.  Effects of Fungal Root Pathogens on the Population Dynamics of Biocontrol Strains of Fluorescent Pseudomonads in the Wheat Rhizosphere , 1991, Applied and environmental microbiology.

[50]  C. Bull Relationship between root colonization and suppression of Gaeumannomyces graminis var. tritici by Pseudomonas fluorescens strains 2-79 , 1991 .

[51]  D. Hartnett,et al.  Competition between Pyrenophora tritici-repentis and Septoria nodorum in the wheat leaf as measured with de Wit replacement series , 1990 .

[52]  G. Défago,et al.  Naturally occurring fluorescent pseudomonads involved in suppression of black root rot of tobacco , 1986 .

[53]  C. R. Howell Suppression of Pythium ultimum-induced damping-off of cotton seedlings by Pseudomonas fluorescens and its antibiotic, pyoluteorin. , 1980 .