Rhizosphere Competence of Wild-Type and Genetically Engineered Pseudomonas brassicacearum Is Affected by the Crop Species.

2,4-Diacetylphloroglucinol (2,4-DAPG)-producing Pseudomonas brassicacearum Q8r1-96 is a highly effective biocontrol agent of take-all disease of wheat. Strain Z30-97, a recombinant derivative of Q8r1-96 containing the phzABCDEFG operon from P. synxantha (formerly P. fluorescens) 2-79 inserted into its chromosome, also produces phenazine-1-carboxylic acid. Rhizosphere population sizes of Q8r1-96, Z30-97, and 2-79, introduced into the soil, were assayed during successive growth cycles of barley, navy bean, or pea under controlled conditions as a measure of the impact of crop species on rhizosphere colonization of each strain. In the barley rhizosphere, Z30-96 colonized less that Q8r1-96 when they were introduced separately, and Q8r1-96 out-competed Z30-96 when the strains were introduced together. In the navy bean rhizosphere, Q8r1-96 colonized better than Z30-97 when the strains were introduced separately. However, both strains had similar population densities when introduced together. Strain Q8r1-96 and Z30-97 colonized the pea rhizosphere equally well when each strain was introduced separately, but Z30-97 out-competed Q8r1-96 when they were introduced together. To our knowledge, this is the first report of a recombinant biocontrol strain of Pseudomonas spp. gaining rhizosphere competitiveness on a crop species. When assessing the potential fate of and risk posed by a recombinant Pseudomonas sp. in soil, both the identity of the introduced genes and the crop species colonized by the recombinant strain need to be considered.

[1]  C. Pieterse,et al.  Unraveling Root Developmental Programs Initiated by Beneficial Pseudomonas spp. Bacteria1[C][W][OA] , 2013, Plant Physiology.

[2]  W. Blankenfeldt,et al.  Recent insights into the diversity, frequency and ecological roles of phenazines in fluorescent Pseudomonas spp. , 2013, Environmental microbiology.

[3]  J. Junaid,et al.  Commercial Biocontrol Agents and Their Mechanism of Action in the Management of Plant Pathogens , 2013 .

[4]  M. Mazzola,et al.  Diversity and natural functions of antibiotics produced by beneficial and plant pathogenic bacteria. , 2012, Annual review of phytopathology.

[5]  Young Cheol Kim,et al.  Comparative Genomics of Plant-Associated Pseudomonas spp.: Insights into Diversity and Inheritance of Traits Involved in Multitrophic Interactions , 2012, PLoS genetics.

[6]  L. Thomashow,et al.  Population Structure and Diversity of Phenazine-1-Carboxylic Acid Producing Fluorescent Pseudomonas spp. from Dryland Cereal Fields of Central Washington State (USA) , 2012, Microbial Ecology.

[7]  T. Paulitz,et al.  Biological control of take-all by fluorescent Pseudomonas spp. from Chinese wheat fields. , 2011, Phytopathology.

[8]  Fang Yang,et al.  Biosynthesis of phloroglucinol compounds in microorganisms—review , 2011, Applied Microbiology and Biotechnology.

[9]  L. Pierson,et al.  Metabolism and function of phenazines in bacteria: impacts on the behavior of bacteria in the environment and biotechnological processes , 2010, Applied Microbiology and Biotechnology.

[10]  W. Blankenfeldt,et al.  Diversity and Evolution of the Phenazine Biosynthesis Pathway , 2009, Applied and Environmental Microbiology.

[11]  B. Lugtenberg,et al.  Plant-growth-promoting rhizobacteria. , 2009, Annual review of microbiology.

[12]  D. Fitzpatrick Lines of Evidence for Horizontal Gene Transfer of a Phenazine Producing Operon into Multiple Bacterial Species , 2009, Journal of Molecular Evolution.

[13]  C. Keel,et al.  Dialogues of root-colonizing biocontrol pseudomonads , 2007, European Journal of Plant Pathology.

[14]  V. Stockwell,et al.  Using Pseudomonas spp. for Integrated Biological Control. , 2007, Phytopathology.

[15]  L. Thomashow,et al.  SELECTING, MONITORING, AND ENHANCING THE PERFORMANCE OF BACTERIAL BIOCONTROL AGENTS: PRINCIPLES, PITFALLS, AND PROGRESS , 2007 .

[16]  B. Landa,et al.  Role of 2,4-Diacetylphloroglucinol-Producing Fluorescent Pseudomonas spp. in the Defense of Plant Roots , 2006 .

[17]  L. Thomashow,et al.  Role of ptsP, orfT, and sss Recombinase Genes in Root Colonization by Pseudomonas fluorescens Q8r1-96 , 2006, Applied and Environmental Microbiology.

[18]  W. Blankenfeldt,et al.  Phenazine compounds in fluorescent Pseudomonas spp. biosynthesis and regulation. , 2006, Annual review of phytopathology.

[19]  L. Pierson,et al.  Quorum Sensing and Phenazines are Involved in Biofilm Formation by Pseudomonas chlororaphis (aureofaciens) Strain 30-84 , 2006, Microbial Ecology.

[20]  F. O'Gara,et al.  Molecular-based strategies to exploit Pseudomonas biocontrol strains for environmental biotechnology applications. , 2006, FEMS microbiology ecology.

[21]  K. K. Pal,et al.  Biological Control of Plant Pathogens , 2006 .

[22]  A. Johansen,et al.  Non-target effects of the microbial control agents Pseudomonas fluorescens DR54 and Clonostachys rosea IK726 in soils cropped with barley followed by sugar beet: a greenhouse assessment , 2005 .

[23]  P. Bakker,et al.  Ascomycete communities in the rhizosphere of field-grown wheat are not affected by introductions of genetically modified Pseudomonas putida WCS358r. , 2005, Environmental microbiology.

[24]  D. Fravel Commercialization and Implementation of Biocontrol 1 , 2005 .

[25]  D. Haas,et al.  Biological control of soil-borne pathogens by fluorescent pseudomonads , 2005, Nature Reviews Microbiology.

[26]  L. Thomashow,et al.  Transformation of Pseudomonas fluorescens with genes for biosynthesis of phenazine-1-carboxylic acid improves biocontrol of rhizoctonia root rot and in situ antibiotic production. , 2004, FEMS microbiology ecology.

[27]  B. Landa,et al.  Minimal changes in rhizobacterial population structure following root colonization by wild type and transgenic biocontrol strains. , 2004, FEMS microbiology ecology.

[28]  D. Hassett,et al.  Pseudomonas aeruginosa Pyocyanin Is Critical for Lung Infection in Mice , 2004, Infection and Immunity.

[29]  B. Landa,et al.  Influence of temperature on plant–rhizobacteria interactions related to biocontrol potential for suppression of fusarium wilt of chickpea , 2004 .

[30]  A. Winding,et al.  Non-target effects of bacterial biological control agents suppressing root pathogenic fungi. , 2004, FEMS microbiology ecology.

[31]  P. Bakker,et al.  Effects of Pseudomonas putida modified to produce phenazine-1-carboxylic acid and 2,4-diacetylphloroglucinol on the microflora of field grown wheat , 2002, Antonie van Leeuwenhoek.

[32]  Jos M. Raaijmakers,et al.  Antibiotic production by bacterial biocontrol agents , 2004, Antonie van Leeuwenhoek.

[33]  Brion Duffy,et al.  Pathogen self-defense: mechanisms to counteract microbial antagonism,. , 2003, Annual review of phytopathology.

[34]  S. Shaukat,et al.  Impact of biocontrol agents Pseudomonas fluorescens CHA0 and its genetically modified derivatives on the diversity of culturable fungi in the rhizosphere of mungbean , 2003, Journal of applied microbiology.

[35]  B. Landa,et al.  Interactions Between Strains of 2,4-Diacetylphloroglucinol-Producing Pseudomonas fluorescens in the Rhizosphere of Wheat. , 2003, Phytopathology.

[36]  B. Ownley,et al.  Identification and Manipulation of Soil Properties To Improve the Biological Control Performance of Phenazine-Producing Pseudomonas fluorescens , 2003, Applied and Environmental Microbiology.

[37]  P. Bakker,et al.  Repeated Introduction of Genetically Modified Pseudomonas putida WCS358r without Intensified Effects on the Indigenous Microflora of Field-Grown Wheat , 2003, Applied and Environmental Microbiology.

[38]  T. Chin-A-Woeng,et al.  Phenazines and their role in biocontrol by Pseudomonas bacteria. , 2003, The New phytologist.

[39]  N. Amarger Genetically modified bacteria in agriculture. , 2002, Biochimie.

[40]  D. Glandorf,et al.  Effects of Pseudomonas putida WCS358r and its genetically modified phenazine producing derivative on the Fusarium population in a field experiment, as determined by 18S rDNA analysis , 2002 .

[41]  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.

[42]  D. Haas,et al.  Fusaric Acid-Producing Strains of Fusarium oxysporum Alter 2,4-Diacetylphloroglucinol Biosynthetic Gene Expression in Pseudomonas fluorescens CHA0 In Vitro and in the Rhizosphere of Wheat , 2002, Applied and Environmental Microbiology.

[43]  B. M. Gardener,et al.  Biological Control of Plant Pathogens: Research, Commercialization, and Application in the USA , 2002 .

[44]  L. Thomashow,et al.  Functional Analysis of Genes for Biosynthesis of Pyocyanin and Phenazine-1-Carboxamide from Pseudomonas aeruginosa PAO1 , 2001, Journal of bacteriology.

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

[46]  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.

[47]  J. Thomas-Oates,et al.  Introduction of the phzH gene of Pseudomonas chlororaphis PCL1391 extends the range of biocontrol ability of phenazine-1-carboxylic acid-producing Pseudomonas spp. strains. , 2001, Molecular plant-microbe interactions : MPMI.

[48]  P. Bakker,et al.  Effect of Genetically Modified Pseudomonas putida WCS358r on the Fungal Rhizosphere Microflora of Field-Grown Wheat , 2001, Applied and Environmental Microbiology.

[49]  M. Girlanda,et al.  Impact of Biocontrol Pseudomonas fluorescens CHA0 and a Genetically Modified Derivative on the Diversity of Culturable Fungi in the Cucumber Rhizosphere , 2001, Applied and Environmental Microbiology.

[50]  A. Winding,et al.  Succession of Indigenous Pseudomonas spp. and Actinomycetes on Barley Roots Affected by the Antagonistic StrainPseudomonas fluorescens DR54 and the Fungicide Imazalil , 2001, Applied and Environmental Microbiology.

[51]  J. Lynch,et al.  Biocontrol of Pythium in the pea rhizosphere by antifungal metabolite producing and non‐producing Pseudomonas strains , 2001, Journal of applied microbiology.

[52]  L. Thomashow,et al.  phzO, a Gene for Biosynthesis of 2-Hydroxylated Phenazine Compounds in Pseudomonas aureofaciens 30-84 , 2001, Journal of bacteriology.

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

[54]  M. Bailey,et al.  Chromosomal insertion of phenazine-1-carboxylic acid biosynthetic pathway enhances efficacy of damping-off disease control by Pseudomonas fluorescens. , 2000, Molecular plant-microbe interactions : MPMI.

[55]  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.

[56]  P. Bakker,et al.  Biocontrol by Phenazine-1-carboxamide-Producing Pseudomonas chlororaphis PCL1391 of Tomato Root Rot Caused by Fusarium oxysporum f. sp. radicis-lycopersici , 1998 .

[57]  A. Boronin,et al.  A Seven-Gene Locus for Synthesis of Phenazine-1-Carboxylic Acid by Pseudomonas fluorescens2-79 , 1998, Journal of bacteriology.

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

[59]  C. Keel,et al.  Influence of biocontrol strain Pseudomonas fluorescens CHA0 and its antibiotic overproducing derivative on the diversity of resident root colonizing pseudomonads , 1997 .

[60]  L. Thomashow,et al.  Frequency of Antibiotic-Producing Pseudomonas spp. in Natural Environments , 1997, Applied and environmental microbiology.

[61]  J. Lynch,et al.  Impact of Field Release of Genetically Modified Pseudomonas fluorescens on Indigenous Microbial Populations of Wheat , 1995, Applied and environmental microbiology.

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

[63]  B. Ownley Influence of in situ and in vitro pH on suppression of Gaeumannomyces graminis var. tritici by Pseudomonas fluorescens 2-79. , 1992 .

[64]  V. de Lorenzo,et al.  Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria , 1990, Journal of bacteriology.

[65]  L. Thomashow,et al.  Role of a phenazine antibiotic from Pseudomonas fluorescens in biological control of Gaeumannomyces graminis var. tritici , 1988, Journal of bacteriology.