Applied Microbiology and Biotechnology

, Volume 68, Issue 5, pp 630–638 | Cite as

Proline-based modulation of 2,4-diacetylphloroglucinol and viable cell yields in cultures of Pseudomonas fluorescens wild-type and over-producing strains

Biotechnological Products and Process Engineering

Abstract

The antifungal compound 2,4-diacetylphloroglucinol (DAPG) is produced in the rhizosphere of wheat by pseudomonad populations responsible for the natural biological control phenomenon known as “take-all decline.” Studies were conducted to elucidate the impact of DAPG and its co-product 2,4,6-trihydroxyacetophenone (THA) on the production of Pseudomonas fluorescens for biological control. Increasing DAPG from 0.1 g/l to 0.5 g/l and THA from 0.05 g/l to 0.5 g/l significantly inhibited the growth and lowered the yield of viable bacteria in liquid cultures. On further examination of these metabolites applied in seed coatings, levels of DAPG and THA exceeding 0.05 mg/g seed significantly reduced wheat germination percentages. The three-way interaction of DAPG, THA, and culture medium ingredients was significant, and greatest seed germination loss (40–50%) was observed when 0.5 mg DAPG and 0.25 mg THA were combined in a coating of 0.5 ml culture medium per gram of seed. Based on the results of Biolog GN microplate, flask, and fermentor screens of C sources, proline was found to optimize the viable cell yields of the P. fluorescens strains tested. The combination of proline with glucose and urea as C and N sources in growth media could be optimized to minimize DAPG production and maximize the vitality of P. fluorescens Q8R1-96 and Q69c-80:miniTn5:phl20 (DAPG over-producer). In production cultures, the proline supply rate offers a potentially useful means to optimize the biological control agent yield and quality.

References

  1. Bochner BR (1978) Device, compostion and method for identifying microorganisms. US patent 4,129,483Google Scholar
  2. Cliquet S, Jackson MA (1999) Influence of culture conditions on production and freeze-drying tolerance of Paecilomyces fumosoroseus blastospores. J Ind Microbiol Biotechnol 23:97–102CrossRefPubMedGoogle Scholar
  3. Cook RJ, Veseth RJ (1991) Wheat health management. APS, St. PaulGoogle Scholar
  4. DaCruz SH, Cilli EM, Emandes JR (2002) Structural complexity of the nitrogen source and influence on yeast growth and fermentation. J Inst Brew 108:54–61Google Scholar
  5. Doudoroff M, Palleroni NJ (1974) Genus I: Pseudomonas. In: Buchanan RE, Gibbons NE (eds) Bergey’s manual of determinative bacteriology. Williams & Wilkins, Baltimore, p. 218Google Scholar
  6. Duffy BK, Défago G (1999) Environmental factors modulating antibiotic and siderophore biosynthesis by Pseudomonas fluorescens biocontrol strains. Appl Environ Microbiol 65:2429–2438PubMedGoogle Scholar
  7. Harrison LA, Letendre L, Kovacevich P, Pierson E, and Weller D (1993) Purification of an antibiotic effective against Gaeumannomyces graminis var. tritici produced by a biocontrol agent, Pseudomonas aureofaciens. Soil Biol Biochem 25:215–221CrossRefGoogle Scholar
  8. Keel C, Schnider U, Maurhofer M, Voisard C, Laville J, Burger U, Wirthner P, Haas D, and Défago G et al. (1992) Suppression of root diseases by Pseudomonas fluorescens CHA0: importance of the bacterial secondary metabolite 2,4-diacetylphloroglucinol. Mol Plant-Microb Interact 5:4–13Google Scholar
  9. Mazzola M, Fujimoto DK, Thomashow LS, and Cook RJ. (1995) Variation in sensitivity of Gaeumannomyces graminis to antibiotics produced by fluorescent Pseudomonad spp. and effect on biological control of take-all of wheat. Appl Environ Microbiol 61:2554–2559Google Scholar
  10. Morita Y, Nakamori S, Takagi H (2003) l-Proline accumulation and freeze tolerance in Saccharomyces cerevisiae are caused by a mutation in the PRO1 gene encoding γ-glutamyl kinase. Appl Environ Microbiol 69:212–219CrossRefPubMedGoogle Scholar
  11. Reddi RK, Khudyakov YP, Borovkov A (1969) Pseudomonas fluorescens strain 26-0, producing phytotoxic substances. Mikrobiologija 38:909–913Google Scholar
  12. Selmer-Olsen E, Pehrson R, Soerhaug T, Birkeland SE (1996) Effect of drying medium on the viability of dried Lactobacillus helveticus CNRZ 303 immobilized in calcium alginate beads. Prog Biotechnol 11:229–235Google Scholar
  13. Shanahan P, O’Sullivan DJ, Simpson P, Glennon JD, and O’Gara F (1992) Isolation of 2,4-diacetylphloroglucinol from a fluorescent Pseudomonad and investigation of physiological parameters influencing its production. Appl Environ Microbiol 58:353–358PubMedGoogle Scholar
  14. Shanahan P, Glennon JD, Crowley JJ, Donnelly DF, O’Gara F (1993) Liquid chromatographic assay of microbially derived phloroglucinol antibiotics for establishing the biosynthetic route to production, and the factors affecting their regulation. Anal Chim Acta 272:271–277CrossRefGoogle Scholar
  15. Slininger PJ, Jackson MA (1992) Nutritional factors regulating growth and accumulation of phenazine 1-carboxylic acid by Pseudomonas fluorescens 2-79. Appl Microbiol Biotechnol 37:388–392CrossRefGoogle Scholar
  16. Slininger PJ, Shea-Wilbur MA (1995) Liquid-culture pH, temperature, and carbon (not nitrogen) source regulate phenazine productivity of the take-all biocontrol agent Pseudomonas fluorescens 2-79. Appl Microbiol Biotechnol 43:794–800CrossRefPubMedGoogle Scholar
  17. Slininger PJ, VanCauwenberge JE, Bothast RJ, Weller DM, Thomashow LS, Cook RJ (1996) Effect of growth culture physiological state, metabolites, and formulation on the viability, phytotoxicity, and efficacy of the take-all biocontrol agent Pseudomonas fluorescens 2-79 stored encapsulated on wheat seeds. Appl Microbiol Biotechnol 45:391–398CrossRefGoogle Scholar
  18. Takagi H, Iwamoto F, Nakamori S (1997) Isolation of freeze-tolerant laboratory strains of Saccharomyces cerevisiae from proline–analog-resistant mutants. Appl Microbiol Biotechnol 47:405–411CrossRefPubMedGoogle Scholar
  19. Takagi H, Sakai K, Morida K, Nakamori S (2000) Proline accumulation by mutation or disruption of the proline oxidase gene improves resistance to freezing and desiccation stresses in Saccharomyces cerevisiae. FEMS Microbiol Lett 184:103–108CrossRefPubMedGoogle Scholar
  20. Thomas KC, Hynes SH, Ingledew WM (1994) Effects of particulate materials and osmoprotectants on very-high-gravity ethanolic fermentation by Saccharomyces cerevisiae. Appl Environ Microbiol 60:1512–1518PubMedGoogle Scholar
  21. USDA (1994) Agriculture fact book. Office of Communications, Washington, D.C.Google Scholar
  22. Vidaver AK (1967) Synthetic and complex media for the rapid detection of fluorescence of phytopathogenic Pseudomonads: effect of the carbon source. Appl Microbiol 15:1523–1524PubMedGoogle Scholar
  23. Weller DM, Zhang BX, Cook RJ (1985) Application of a rapid screening test for selection of bacteria suppressive to take-all of wheat. Plant Dis 69:710–713Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  1. 1.Agricultural Research Service, National Center for Agricultural Utilization ResearchUnited States Department of AgriculturePeoriaUSA

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