Pseudomonas fluorescens

Pseudomonas fluorescens is a common Gram-negative, rod-shaped bacterium.[1] It belongs to the Pseudomonas genus; 16S rRNA analysis has placed P. fluorescens in the P. fluorescens group within the genus,[2] to which it lends its name.

Pseudomonas fluorescens
Pseudomonas fluorescens under white light
The same plate under UV light
Scientific classification
Domain: Bacteria
Phylum: Proteobacteria
Class: Gammaproteobacteria
Order: Pseudomonadales
Family: Pseudomonadaceae
Genus: Pseudomonas
Species group: Pseudomonas fluorescens group
Species:
P. fluorescens
Binomial name
Pseudomonas fluorescens
(Flügge 1886)
Migula, 1895
Type strain
ATCC 13525

CCUG 1253
CCEB 546
CFBP 2102
CIP 69.13
DSM 50090
JCM 5963
LMG 1794
NBRC 14160
NCCB 76040
NCIMB 9046
NCTC 10038
NRRL B-14678
VKM B-894

Synonyms

Bacillus fluorescens liquefaciens Flügge 1886
Bacillus fluorescens Trevisan 1889
Bacterium fluorescens (Trevisan 1889) Lehmann and Neumann 1896
Liquidomonas fluorescens (Trevisan 1889) Orla-Jensen 1909
Pseudomonas lemonnieri (Lasseur) Breed 1948
Pseudomonas schuylkilliensis Chester 1952
Pseudomonas washingtoniae (Pine) Elliott

General characteristics

Pseudomonas fluorescens has multiple flagella. It has an extremely versatile metabolism, and can be found in the soil and in water. It is an obligate aerobe, but certain strains are capable of using nitrate instead of oxygen as a final electron acceptor during cellular respiration.

Optimal temperatures for growth of P. fluorescens are 25–30°C. It tests positive for the oxidase test. It is also a nonsaccharolytic bacterial species.

Heat-stable lipases and proteases are produced by P. fluorescens and other similar pseudomonads.[3] These enzymes cause milk to spoil, by causing bitterness, casein breakdown, and ropiness due to production of slime and coagulation of proteins.[4][5]

The name

The word Pseudomonas means false unit, being derived from the Greek words pseudēs (Greek: ψευδής – false) and monas (Latin: monas, from Greek: μονάς – a single unit). The word was used early in the history of microbiology to refer to germs. The specific name fluorescens refers to the microbe's secretion of a soluble fluorescent pigment called pyoverdin, which is a type of siderophore.[6]

Genome-sequencing projects

The genomes of P. fluorescens strains SBW25,[7] Pf-5[8] and PfO-1[9] have been sequenced.

Interactions with Dictyostelium

There are two strains of Pseudomonas fluorescens associated with Dictyostelium discoideum. One strain serves as a food source and the other strain does not. The main genetic difference between these two strains is a mutation of the global activator gene called gacA. This gene plays a key role in gene regulation; when this gene is mutated in the nonfood bacterial strain, it is transformed into a food bacterial strain.[10]

Biocontrol properties

Some P. fluorescens strains (CHA0 or Pf-5, for example) present biocontrol properties, protecting the roots of some plant species against parasitic fungi such as Fusarium or the oomycete Pythium, as well as some phytophagous nematodes.[11]

It is not clear exactly how the plant growth-promoting properties of P. fluorescens are achieved; theories include:

  • The bacteria might induce systemic resistance in the host plant, so it can better resist attack by a true pathogen.
  • The bacteria might outcompete other (pathogenic) soil microbes, e.g., by siderophores, giving a competitive advantage at scavenging for iron.
  • The bacteria might produce compounds antagonistic to other soil microbes, such as phenazine-type antibiotics or hydrogen cyanide.

To be specific, certain P. fluorescens isolates produce the secondary metabolite 2,4-diacetylphloroglucinol (2,4-DAPG), the compound found to be responsible for antiphytopathogenic and biocontrol properties in these strains.[12] The phl gene cluster encodes factors for 2,4-DAPG biosynthesis, regulation, export, and degradation. Eight genes, phlHGFACBDE, are annotated in this cluster and conserved organizationally in 2,4-DAPG-producing strains of P. fluorescens. Of these genes, phlD encodes a type III polyketide synthase, representing the key biosynthetic factor for 2,4-DAPG production. PhlD shows similarity to plant chalcone synthases and has been theorized to originate from horizontal gene transfer.[13] Phylogenetic and genomic analysis, though, has revealed that the entire phl gene cluster is ancestral to P. fluorescens, many strains have lost the capacity, and it exists on different genomic regions among strains.[14]

Some experimental evidence supports all of these theories, in certain conditions; a good review of the topic is written by Haas and Defago.[15]

Several strains of P. fluorescens, such as Pf-5 and JL3985, have developed a natural resistance to ampicillin and streptomycin.[16] These antibiotics are regularly used in biological research as a selective pressure tool to promote plasmid expression.

The strain referred to as Pf-CL145A has proved itself a promising solution for the control of invasive zebra mussels and quagga mussels (Dreissena). This bacterial strain is an environmental isolate capable of killing >90% of these mussels by intoxication (i.e., not infection), as a result of natural product(s) associated with their cell walls, and with dead Pf-145A cells killing the mussels equally as well as live cells.[17] Following ingestion of the bacterial cells mussel death occurs following lysis and necrosis of the digestive gland and sloughing of stomach epithelium.[18] Research to date indicates very high specificity to zebra and quagga mussels, with low risk of nontarget impact.[19] Pf-CL145A has now been commercialized under the product name Zequanox, with dead bacterial cells as its active ingredient.

Recent results showed the production of the phytohormone cytokinin by P. fluorescens strain G20-18 to be critical for its biocontrol activity by activating plant resistance.[20]

Medical properties

By culturing P. fluorescens, mupirocin (an antibiotic) can be produced, which has been found to be useful in treating skin, ear, and eye disorders.[21] Mupirocin free acid and its salts and esters are agents currently used in creams, ointments, and sprays as a treatment of methicillin-resistant Staphylococcus aureus infection.

Pseudomonas fluorescens demonstrates hemolytic activity, and as a result, has been known to infect blood transfusions.[22]

Disease

Pseudomonas fluorescens is an unusual cause of disease in humans, and usually affects patients with compromised immune systems (e.g., patients on cancer treatment). From 2004 to 2006, an outbreak of P. fluorescens in the United States involved 80 patients in six states. The source of the infection was contaminated heparinized saline flushes being used with cancer patients.[23]

Pseudomonas fluorescens is also a known cause of fin rot in fish.

Metabolism

Pseudomonas fluorescens produces phenazine, phenazine carboxylic acid,[24] 2,4-diacetylphloroglucinol[25] and the MRSA-active antibiotic mupirocin.[26]

Biodegradation capacities

4-Hydroxyacetophenone monooxygenase is an enzyme found in P. fluorescens that transforms piceol, NADPH, H+, and O2 into 4-hydroxyphenyl acetate, NADP+, and H2O.

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References
  1. Palleroni, N.J. (1984) Pseudomonadaceae. Bergey's Manual of Systematic Bacteriology. Krieg, N. R. and Holt J. G. (editors) Baltimore: The Williams and Wilkins Co., pg. 141 – 199
  2. Anzai; Kim, H; Park, JY; Wakabayashi, H; Oyaizu, H; et al. (Jul 2000). "Phylogenetic affiliation of the pseudomonads based on 16S rRNA sequence". Int J Syst Evol Microbiol. 50 (4): 1563–89. doi:10.1099/00207713-50-4-1563. PMID 10939664.
  3. Frank, J.F. 1997. Milk and dairy products. In Food Microbiology, Fundamentals and Frontiers, ed. M.P. Doyle, L.R. Beuchat, T.J. Montville, ASM Press, Washington, p. 101.
  4. Jay, J.M. 2000. Taxonomy, role, and significance of microorganisms in food. In Modern Food Microbiology, Aspen Publishers, Gaithersburg MD, p. 13.
  5. Ray, B. 1996. Spoilage of Specific food groups. In Fundamental Food Microbiology, CRC Press, Boca Raton FL, p. 220. I
  6. C D Cox and P Adams (1985) Infection and Immunity 48(1): 130–138
  7. Pseudomonas fluorescens
  8. "Pseudomonas fluorescens Pf-5 Genome Page". Archived from the original on 2009-06-28. Retrieved 2009-04-23.
  9. "Pseudomonas fluorescens PfO-1 Genome Page". Archived from the original on 2009-06-24. Retrieved 2009-04-23.
  10. Stallforth, Pierre; Brock, Debra A.; Cantley, Alexandra M.; Tian, Xiangjun; Queller, David C.; Strassmann, Joan E.; Clardy, Jon (2013-09-03). "A bacterial symbiont is converted from an inedible producer of beneficial molecules into food by a single mutation in the gacA gene". Proceedings of the National Academy of Sciences of the United States of America. 110 (36): 14528–14533. doi:10.1073/pnas.1308199110. ISSN 0027-8424. PMC 3767522. PMID 23898207.
  11. Haas, D.; Keel, C. (2003). "Regulation of antibiotic production in root-colonizing Pseudomonas spp. and relevance for biological control of plant disease". Annual Review of Phytopathology. 41: 117–153. doi:10.1146/annurev.phyto.41.052002.095656. PMID 12730389.
  12. Bangera M. G.; Thomashow L. S. (1999). "Identification and characterization of a gene cluster for synthesis of the polyketide antibiotic 2,4-diacetylphloroglucinol from pseudomonas fluorescens q2-87". Journal of Bacteriology. 181 (10): 3155–3163. doi:10.1128/JB.181.10.3155-3163.1999.
  13. Bangera M. G.; Thomashow L. S. (1999). "Identification and characterization of a gene cluster for synthesis of the polyketide antibiotic 2,4-diacetylphloroglucinol from pseudomonas fluorescens q2-87". Journal of Bacteriology. 181 (10): 3155–3163. doi:10.1128/JB.181.10.3155-3163.1999.
  14. Moynihan J. A.; Morrissey J. P.; Coppoolse E. R.; Stiekema W. J.; O'Gara F.; Boyd E. F. (2009). "Evolutionary history of the phl gene cluster in the plant-associated bacterium pseudomonas fluorescens". Applied and Environmental Microbiology. 75 (7): 2122–2131. doi:10.1128/aem.02052-08. PMC 2663185. PMID 19181839.
  15. Haas, D; Defago, G (2005). "Biological control of soil-borne pathogens by fluorescent pseudomonads". Nature Reviews Microbiology. 3 (4): 307–19. doi:10.1038/nrmicro1129. PMID 15759041.
  16. Alain Sarniguet; et al. (1995). "The sigma factor σs affects antibiotic production and biological control activity of Pseudomonas fluorescens Pf-5". Proc. Natl. Acad. Sci. U.S.A. 92 (26): 12255–12259. doi:10.1073/pnas.92.26.12255. PMC 40335. PMID 8618880.
  17. Molloy, D. P., Mayer, D. A., Gaylo, M. J., Morse, J. T., Presti, K. T., Sawyko, P. M., Karatayev, A. Y., Burlakova, L. E., Laruelle, F., Nishikawa, K. C., Griffin, B. H. 2013. Pseudomonas fluorescens strain CL145A – A biopesticide for the control of zebra and quagga mussels (Bivalvia: Dreissenidae). J. Invertebr. Pathol. 113(1):104–114.
  18. Molloy, D. P., Mayer, D. A., Giamberini, L., and Gaylo, M. J. 2013. Mode of action of Pseudomonas fluorescens strain CL145A, a lethal control agent of dreissenid mussels (Bivalvia: Dreissenidae). J. Invertebr. Pathol. 113(1):115–121.
  19. Molloy, D. P.; Mayer, D. A.; Gaylo, M. J.; Burlakova, L. E.; Karatayev, A. Y.; Presti, K. T.; Sawyko, P. M.; Morse, J. T.; Paul, E. A. (2013). "Non-target trials with Pseudomonas fluorescens strain CL145A, a lethal control agent of dreissenid mussels (Bivalvia: Dreissenidae)". Manag. Biol. Invasions. 4 (1): 71–79. doi:10.3391/mbi.2013.4.1.09.
  20. Großkinsky DK, Tafner R, Moreno MV, Stenglein SA, García de Salamone IE, Nelson LM, Novák O, Strnad M, van der Graaff E, Roitsch T (2016). "Cytokinin production by Pseudomonas fluorescens G20-18 determines biocontrol activity against Pseudomonas syringae in Arabidopsis". Scientific Reports. 6: 23310. doi:10.1038/srep23310. PMC 4794740. PMID 26984671.
  21. Bactroban
  22. Gibb AP, Martin KM, Davidson GA, Walker B, Murphy WG (1995). "Rate of growth of Pseudomonas fluorescens in donated blood". Journal of Clinical Pathology. 48 (8): 717–8. doi:10.1136/jcp.48.8.717. PMC 502796. PMID 7560196.
  23. Gershman MD, Kennedy DJ, Noble-Wang J, et al. (2008). "Multistate outbreak of Pseudomonas fluorescens bloodstream infection after exposure to contaminated heparinized saline flush prepared by a compounding pharmacy". Clin Infect Dis. 47 (11): 1372–1379. doi:10.1086/592968. PMID 18937575.
  24. Mavrodi, D.V.; Ksenzenko, V. N.; Bonsall, R. F.; Cook, R. J.; Boronin, A. M.; Thomashow, L. S. (1998). "A seven-gene locus for synthesis of phenazine-1-carboxylic acid by Pseudomonas fluorescens 2–79". J. Bacteriol. 180 (9): 2541–2548. doi:10.1128/JB.180.9.2541-2548.1998. PMC 107199. PMID 9573209.
  25. Achkar, Jihane; Xian, Mo; Zhao, Huimin; Frost, J. W. (2005). "Biosynthesis of Phloroglucinol". J. Am. Chem. Soc. 127 (15): 5332–5333. doi:10.1021/ja042340g. PMID 15826166.
  26. Fuller, AT; Mellows, G; Woolford, M; Banks, GT; Barrow, KD; Chain, EB (1971). "Pseudomonic acid: an antibiotic produced by Pseudomonas fluorescens". Nature. 234 (5329): 416–417. doi:10.1038/234416a0. PMID 5003547.

Further reading

Appanna, Varun P.; Auger, Christopher; Thomas, Sean C.; Omri, Abdelwahab (13 June 2014). "Fumarate metabolism and ATP production in Pseudomonas fluorescens exposed to nitrosative stress". Antonie van Leeuwenhoek. 106 (3): 431–438. doi:10.1007/s10482-014-0211-7. PMID 24923559.

Cabrefiga, J.; Frances, J.; Montesinos, E.; Bonaterra, A. (1 October 2014). "Improvement of a dry formulation of Pseudomonas fluorescens EPS62e for fire blight disease biocontrol by combination of culture osmoadaptation with a freeze-drying lyoprotectant". Journal of Applied Microbiology. 117 (4): 1122–1131. doi:10.1111/jam.12582. PMID 24947806. Retrieved 2 November 2014.

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