Biological control of Rhizoctonia root rot on bean by phenazine- and cyclic lipopeptide-producing Pseudomonas CMR12a.

Pseudomonas CMR12a was previously selected as an efficient biocontrol strain producing phenazines and cyclic lipopeptides (CLPs). In this study, biocontrol capacity of Pseudomonas CMR12a against Rhizoctonia root rot of bean and the involvement of phenazines and CLPs in this ability were tested. Two different anastomosis groups (AGs) of Rhizoctonia solani, the intermediately aggressive AG 2-2 and the highly aggressive AG 4 HGI, were included in growth-chamber experiments with bean plants. The wild-type strain CMR12a dramatically reduced disease severity caused by both R. solani AGs. A CLP-deficient and a phenazine-deficient mutant of CMR12a still protected bean plants, albeit to a lesser extent compared with the wild type. Two mutants deficient in both phenazine and CLP production completely lost their biocontrol activity. Disease-suppressive capacity of CMR12a decreased after washing bacteria before application to soil and thereby removing metabolites produced during growth on plate. In addition, microscopic observations revealed pronounced branching of hyphal tips of both R. solani AGs in the presence of CMR12a. More branched and denser mycelium was also observed for the phenazine-deficient mutant; however, neither the CLP-deficient mutant nor the mutants deficient in both CLPs and phenazines influenced hyphal growth. Together, results demonstrate the involvement of phenazines and CLPs during Pseudomonas CMR12a-mediated biocontrol of Rhizoctonia root rot of bean.

[1]  L. Vanhaecke,et al.  N-Acylhomoserine lactone quorum-sensing signalling in antagonistic phenazine-producing Pseudomonas isolates from the red cocoyam rhizosphere. , 2011, Microbiology.

[2]  M. Höfte,et al.  Rhizoctonia spp. causing root and hypocotyl rot in Phaseolus vulgaris in Cuba , 2010 .

[3]  M. Höfte,et al.  Biosurfactants in plant-Pseudomonas interactions and their importance to biocontrol. , 2009, Environmental microbiology reports.

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

[5]  S. Sindhu.  Biological Control of Soilborne Plant Pathogens with Rhizosphere Bacteria , 2009 .

[6]  K. Rabaey,et al.  Phenazines and biosurfactants interact in the biological control of soil-borne diseases caused by Pythium spp. , 2008, Environmental microbiology.

[7]  W. Verstraete,et al.  Metabolites produced by Pseudomonas sp. enable a Gram-positive bacterium to achieve extracellular electron transfer , 2008, Applied Microbiology and Biotechnology.

[8]  M. Höfte,et al.  Biosurfactants are involved in the biological control of Verticillium microsclerotia by Pseudomonas spp. , 2007, Journal of applied microbiology.

[9]  P. de Vos,et al.  Characterization of CMR5c and CMR12a, novel fluorescent Pseudomonas strains from the cocoyam rhizosphere with biocontrol activity , 2007, Journal of applied microbiology.

[10]  M. Höfte,et al.  Role of the cyclic lipopeptide massetolide A in biological control of Phytophthora infestans and in colonization of tomato plants by Pseudomonas fluorescens. , 2007, The New phytologist.

[11]  D. Weller.  Pseudomonas biocontrol agents of soilborne pathogens: looking back over 30 years. , 2007, Phytopathology.

[12]  D. Newman,et al.  The phenazine pyocyanin is a terminal signalling factor in the quorum sensing network of Pseudomonas aeruginosa , 2006, Molecular microbiology.

[13]  I. de Bruijn,et al.  Cyclic lipopeptide production by plant-associated Pseudomonas spp.: diversity, activity, biosynthesis, and regulation. , 2006, Molecular plant-microbe interactions : MPMI.

[14]  G. O’Toole,et al.  Saccharomyces cerevisiae-Based Molecular Tool Kit for Manipulation of Genes from Gram-Negative Bacteria , 2006, Applied and Environmental Microbiology.

[15]  M. Höfte,et al.  Control of Phytophthora cryptogea in the hydroponic forcing of witloof chicory with the rhamnolipid‐based biosurfactant formulation PRO1 , 2005 .

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

[17]  C. Ryu,et al.  Induced Systemic Resistance and Promotion of Plant Growth by Bacillus spp. , 2004, Phytopathology.

[18]  M. Kowsari,et al.  Pectic zymogram variation and pathogenicity of Rhizoctonia solani AG-4 to bean (Phaseolus vulgaris) isolates in Isfahn, Iran , 2004, Mycopathologia.

[19]  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.

[20]  C. Campion,et al.  Anastomosis Groups, Pathogenicity and Sensitivity to Fungicides of Rhizoctonia solani Isolates Collected on Potato Crops in France , 2003, European Journal of Plant Pathology.

[21]  M. Höfte,et al.  Phenazines are Involved in Biocontrol of Pythium myriotylum on Cocoyam by Pseudomonas aeruginosa PNA1 , 2001, European Journal of Plant Pathology.

[22]  L. Panella,et al.  A Greenhouse Test for Screening Sugar Beet (Beta Vulgaris) for Resistance to Rhizoctonia Solani , 2001, European Journal of Plant Pathology.

[23]  T. V. van Beek,et al.  Biochemical, Genetic, and Zoosporicidal Properties of Cyclic Lipopeptide Surfactants Produced by Pseudomonas fluorescens , 2003, Applied and Environmental Microbiology.

[24]  G. Lamers,et al.  Interactions in the tomato rhizosphere of two Pseudomonas biocontrol strains with the phytopathogenic fungus Fusarium oxysporum f. sp. radicis-lycopersici. , 2003, Molecular plant-microbe interactions : MPMI.

[25]  S. Beatson,et al.  Identification of type II and type III pyoverdine receptors from Pseudomonas aeruginosa. , 2003, Microbiology.

[26]  O. Nybroe,et al.  Surface motility in Pseudomonas sp. DSS73 is required for efficient biological containment of the root-pathogenic microfungi Rhizoctonia solani and Pythium ultimum. , 2003, Microbiology.

[27]  J. Sørensen,et al.  Antibiotic and Biosurfactant Properties of Cyclic Lipopeptides Produced by Fluorescent Pseudomonas spp. from the Sugar Beet Rhizosphere , 2002, Applied and Environmental Microbiology.

[28]  S. Kuninaga,et al.  Hyphal Anastomosis Reactions, rDNA-Internal Transcribed Spacer Sequences, and Virulence Levels Among Subsets of Rhizoctonia solani Anastomosis Group-2 (AG-2) and AG-BI. , 2002, Phytopathology.

[29]  S. Heeb,et al.  Regulatory roles of the GacS/GacA two-component system in plant-associated and other gram-negative bacteria. , 2001, Molecular plant-microbe interactions : MPMI.

[30]  J. Sørensen,et al.  Pseudomonas fluorescens DR54 Reduces Sclerotia Formation, Biomass Development, and Disease Incidence of Rhizoctonia solani Causing Damping-Off in Sugar Beet , 2001, Microbial Ecology.

[31]  S. Babu,et al.  Induction of systemic resistance in rice against sheath blight disease by Pseudomonas fluorescens , 2001 .

[32]  E. Ron,et al.  Natural roles of biosurfactants. , 2001, Environmental microbiology.

[33]  J. Sørensen,et al.  Structure, production characteristics and fungal antagonism of tensin – a new antifungal cyclic lipopeptide from Pseudomonas fluorescens strain 96.578 , 2000, Journal of applied microbiology.

[34]  J. Sørensen,et al.  Vital fluorescent stains for detection of stress in Pythium ultimum and Rhizoctonia solani challenged with viscosinamide from Pseudomonas fluorescens DR54 , 1999 .

[35]  H. De Greve,et al.  Identification and molecular characterization of a novel Salmonella enteritidis pathogenicity islet encoding an ABC transporter , 1999, Molecular microbiology.

[36]  J. Sørensen,et al.  Viscosinamide, a new cyclic depsipeptide with surfactant and antifungal properties produced by Pseudomonas fluorescens DR54 , 1999, Journal of Applied Microbiology.

[37]  D. Weller,et al.  Effect of Population Density of Pseudomonas fluorescens on Production of 2,4-Diacetylphloroglucinol in the Rhizosphere of Wheat. , 1999, Phytopathology.

[38]  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 .

[39]  Y. Bashan.  INOCULANTS OF PLANT GROWTH-PROMOTING BACTERIA FOR USE IN AGRICULTURE , 1998 .

[40]  H. Schweizer,et al.  A broad-host-range Flp-FRT recombination system for site-specific excision of chromosomally-located DNA sequences: application for isolation of unmarked Pseudomonas aeruginosa mutants. , 1998, Gene.

[41]  M. Stanghellini,et al.  BIOSURFACTANTS: Their Identity and Potential Efficacy in the Biological Control of Zoosporic Plant Pathogens. , 1997, Plant disease.

[42]  J. Handelsman,et al.  Biocontrol of Soilborne Plant Pathogens. , 1996, The Plant cell.

[43]  U. Gisi,et al.  Chemical Control of Rhizoctonia Species , 1996 .

[44]  B. Sneh,et al.  Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control , 1996, Springer Netherlands.

[45]  T. Mew,et al.  Isolation and identification of antifungal metabolites produced by rice-associated antagonistic Pseudomonas spp. , 1995 .

[46]  Schweizer Hd.  Small broad-host-range gentamycin resistance gene cassettes for site-specific insertion and deletion mutagenesis. , 1993 .

[47]  P. Lipps,et al.  Anastomosis grouping and variation in virulence among isolates of Rhizoctonia solani associated with dry bean and soybean in Ohio and Zaire , 1993 .

[48]  H. D. Schweizer.  Small broad-host-range gentamycin resistance gene cassettes for site-specific insertion and deletion mutagenesis. , 1993, BioTechniques.

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

[50]  J. Trevors,et al.  A drop-collapsing test for screening surfactant-producing microorganisms , 1991 .

[51]  V. de Lorenzo,et al.  Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria , 1990, Journal of bacteriology.

[52]  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.

[53]  Teri,et al.  Molecular Cloning A Laboratory Manual Second Edition Sambrook , 1989 .

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

[55]  J. M. Turner,et al.  Occurrence, biochemistry and physiology of phenazine pigment production. , 1986, Advances in microbial physiology.

[56]  D. Hanahan.  Studies on transformation of Escherichia coli with plasmids. , 1983, Journal of molecular biology.

[57]  D. Helinski,et al.  Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[58]  C. R. Howell.  Control of rhizoctonia solani on cotton seedlings with Pseudomonas fluorescens and with an antibiotic produced by the bacterium. , 1979 .

[59]  H. Boyer,et al.  A complementation analysis of the restriction and modification of DNA in Escherichia coli. , 1969, Journal of molecular biology.

[60]  King Eo,et al.  Two simple media for the demonstration of pyocyanin and fluorescin. , 1954 .

[61]  E. King,et al.  Two simple media for the demonstration of pyocyanin and fluorescin. , 1954, The Journal of laboratory and clinical medicine.