Pseudomonas fluorescens

Pseudomonas fluorescens is a common Gram-negative, rod-shaped bacterium. It belongs to the Pseudomonas genus; 16S rRNA analysis has placed P. fluorescens in the P. fluorescens group [2] within the genus, to which it lends its name.
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General characteristics
P. 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 Pseudomonas fluorescens are 25-30 degrees Celsius. It tests positive for the oxidase test. Pseudomonas fluorescens is also a nonsaccharolytic bacteria. Heat-stable lipases and proteases are produced by Pseudomonas fluorescens and other [3] similar pseudomonads. These enzymes cause milk to spoil, by causing bitterness, casein breakdown, and ropiness due to production of slime and coagulation of proteins.

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

sequencing projects
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The genomes of P. fluorescens strains SBW25, [edit]Biocontrol

Pf-5 and PfO-1 have been sequenced.

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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 Pythium, as well as some [10] phytophagous nematodes. 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 phenazinetype antibiotics or hydrogen cyanide

To be specific, certain P. fluorescens isolates produce the secondary metabolite 2,4diacetylphloroglucinol (2,4-DAPG), the compound found to be responsible for antiphytopathogenic [11] and biocontrol properties in these strains. 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 2,4DAPG production. PhlD shows similarity to plant chalcone synthases and has been theorized to

originate from horizontal gene transfer. But phylogenetic and genomic analysis has revealed that the entire phl gene cluster is ancestral to P. fluorescens, many strains have lost the capacity, and it [13] exists on different genomic regions among strains. There is experimental evidence to support all of these theories, in certain conditions; a good review of [14] the topic is written by Haas and Defago. The strain referred to as Pf-CL145A has proved itself a promising solution to the invasive Dreissena( zebra and quagga) mussels. The strain of the bacteria produces toxins that destroy the digestive system of the mussels and produces a >90% kill rate. Several strains of "P. Fluorescens", such as Pf-5 and JL3985, have developed a natural resistance [15] to ampicillin and streptomycin. These antibiotics are regularly used in biological research as a selective pressure tool to promote plasmid expression. [edit]Medical

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Properties

By culturing Pseudomonas fluorescens, Mupirocin (an antibiotic) can be produced, which has been [16] found to be useful in treating skin, ear, and eye disorders. Mupirocin free acid and its salts and esters are agents currently used in creams, ointments, and sprays as a treatment of Methicillinresistant Staphylococcus aureus (MRSA) infection. P. fluorescens demonstrates hemolytic activity and, as a result, has been known to infect blood [17] transfusions. It is also used in milk to make yogurt.
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United States Patents: 6489358, 4873012, 6156792 [edit]Disease P. 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, there was an outbreak of P. fluorescens in the United States, involving 80 patients in six states. The source of the [18] infection was contaminated heparinized saline flushes being used with cancer patients. [edit]Biodegradation

capacities

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

Description and significance

Pseudomonas fluorescens are Gram-negative rod shaped bacteria that inhabit soil, plants, and water surfaces.(2) The optimum growth temperature is between 25-30 degrees Celsius (10). The Pf-5 strain resides in the plants rhizosphere and produces a variety of secondary metabolites including antibiotics against soil borne plant pathogens.(4) Pseudomonas fluorescens PFO-1 is well adapted to the soil where it was first isolated in agricultural soil.(1) Pseudomonas fluorescens strain SBW25 grow on plant leaves and roots where they can contribute to plant growth. Soluble, green fluorescent pigments are produced when the iron concentration is low. The significance of these organisms have increased because of their ability to degrade various pollutants and their use as bio-control against pathogens.(2) Sequencing the genome provided further information of its environmental interaction ands its metabolic capabilities, which can be used against agricultural disease control (1). Pseudomonas fluorescens is interesting and important to study because it produces a particular

antibiotic (Mupirocin) which has been proven effective in treating certain kinds of skin, ear, and eye disorders (10)

Ecology
Pseudomonas fluorescens are commensal species with plants, allowing plants to attain key nutrients, degrading pollutants, and suppressing pathogens via antibiotic productions. These microbes produce secondary metabolites that suppress plant disease and signal gene expression to neighboring cells inhabiting the rhizosphere. Pseudomonas also use siderophores from other microorganism to obtain iron which increases their survival in iron-limited environments. Plants provide these organisms with nutrients and shelter against stressful environments. (4) One of many byproducts of plant cells include active oxygen such as superoxide which are toxic to microbes. Rhizosphere bacteria such as P. fluorescens possess superoxide dismutases to convert superoxide to hydrogen peroxide and catalases to convert peroxide to water. The presence of these enzymes contribute to Pseudomonas fluorescenss tolerance to oxidative stress. (4)

Pathology
Despite their commensal nature, Pseudomonas fluorescens are nonpathogenic and lack virulence factors of other plant pathogens. In Pseudomonas fluroescens Pf-5, enzymes that degrade plant cell walls and their components such as cellulase, pectinase, or pectin lyase are not present. However, it is capable of breaking down some plant-derived carbohydrates, fatty acids, and oils and can hydrolyze proteins causing the spoilage of milk, meat, and fish. It has also been found to be an opportunistic pathogen in immune compromised fish like Koi which are commonly kept in backyard garden ponds. There are other species of Pseudomonas that are pathogens. Pseudomonas syringae is a plant pathogen that impacts food and biomass production. Diseases include bacterial specks on tomatoes and leaves which result in stunted growth and halo blight of beans. These microbes can spread by rain and are seed-borne pathogens(4,6) Although Pseudomonas fluorescens typically has a low level of virulence, in 1997 four patients at the National Taiwan University Hospital developed Pseudomonas fluorescens bacteremia. These patients had been treated in the chemotherapy room and had begun exhibiting symptoms such as fevers and chills. Eight cultures were isolated from catheters and from the blood of the four patients. All isolates were identified as Pseudomonas fluorescens (11).

Pseudomonas aeruginosa are opportunistic human pathogens that are one of the main causes of human infections. These microbes live in diverse environments including soils, marshes, as well as plant and animal tissues, which show their nutritional versatility. Their resistance to antibiotics have made them dangerous pathogens. Pseudomonas aeruginosa are commonly seen in the lungs, especially those with cystic fibrosis. They are also present in urinary-tract infections, burn victims, and patients on respirators with hospital-acquired pneumonia. (5)

Application to Biotechnology
Studies done on Pseudmonas fluorescens have shown the microbes potential benefit in bioremediation against several strains of plant pathogens. The results of the experiment showed that at high concentrations all five strains of Pseudomonas fluorescens tested inhibit spore production by pathogenic plant fungus. Fungi such as Alternaria cajani and Curvularia lunata grow on plant surfaces causing disease and death of the plant. Plant treatment with Pseudomonas fluorescens can prevent these fungi from growing and spreading through spore production. Pseudomonas fluorescens grows

at an optimum temperature of 25 degrees Celsius but can also survive in temperatures as low as 0 degrees C. Therefore, it is rarely pathogenic in humans making it an effective microbe for treating crops since it is not able to survive in the human body. Pseudomonas species are effective against mold causing disease in produce such as apples and pears. This and further studies of Pseudomonas fluorescens will determine its effectiveness an alternative to chemical fungicides.(13) Production of secondary metabolites play an important role in plant disease suppression. Antibiotics such as pyrrolnitrin, pyoluteorin, and 2,4-diacetylphloroglucinol that inhibit phytopathogen growth are produced by Pseudomonas fluorescens Pf-5. Diseases from Rhizoctonia solani and Pythium ultimum that affect cotton plants are inhibited by this strain. Pseudomonas fluorescens produces hydrogen cyanide and the siderophores pyocheline and pyoverdine which it uses to outcompete with many pathogenic bacteria for iron necessary for growth and suppress pathogens in the rhizosphere. The bacteria's degrading ability has been applied to pollutants such as styrene, TNT and, polycyclic aromatic hydrocarbons.(4-6) Pseudomonas fluorescens produce exopolysaccharides which are used for protection against bacteriophages or dehydration as well as for defense against the host immune system. Polysaccharides are being used within the food, chemical, and agricultural industries.(3)