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Chapter 29 : Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy
Authors:
Jean-Marie Meyer1,
Valérie A. Geoffroy1
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Affiliations:
1: Laboratoire de Microbiologie et Génétique, Université Louis-Pasteur-CNRS, FRE 2326, 67000 Strasbourg, France
Content Type: Monograph
Publication Year:
2004
Category: Bacterial Pathogenesis; Clinical Microbiology
Book DOI:
10.1128/9781555816544
Chapter DOI:
10.1128/9781555816544.ch29
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Abstract:
Siderophore-based methods could be successfully used for bacterial identification. Preliminary data for other bacteria (nonfluorescent Pseudomonas species, Burkholderia species, rhizobia, and bradyrhizobia, as well as enterobacteria, mycobacteria, and Aeromonas species) strongly indicate that any siderophore could be used as a taxonomic marker for its producing bacteria, which means that the emerging concept of siderotyping as a taxonomic tool could be applied to a large part of the microbial world. Nonfluorescent siderophores produced by fluorescent pseudomonads, together with pyoverdines, e.g., pyochelin, salicylic acid, or quinolobactin, and other compounds such as those produced by nonfluorescent pseudomonads, e.g., ornibactins, cepabactin, or desferriferrioxamines, are revealed by an overlay of 1% melted agarose in CAS reagent, which reveals siderophores as yellow to pink spots appearing at the surface of the gel. It is clear that siderotyping could be very useful for bacterial identification and Pseudomonas taxonomy: following the recognition of the type of pyoverdine it produces, a taxonomically undefined fluorescent pseudomonad is classified in a corresponding siderovar. The methods of siderotyping are very fast and easy to perform. The method has already proved its efficiency in the bacterial identification of fluorescent Pseudomonas species and in the detection of new species. Depending on the specificity level expressed by the pyoverdine, it should thus be possible to achieve, during a single experimental step, both isolation and identification of a fluorescent Pseudomonas strain.
Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
Figures
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy
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Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
/content/10.1128/9781555816544.chap29.fig29-1
FIGURE 1
Structure of the P. costantinii pyoverdine. Reprinted from Fernandez et al., 2001, with permission from the publisher.
10.1128/9781555816544/fig29-1_thmb.gif
10.1128/9781555816544/fig29-1.gif
FIGURE 1
Structure of the P. costantinii pyoverdine. Reprinted from Fernandez et al., 2001, with permission from the publisher.
Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
/content/10.1128/9781555816544.chap29.fig29-2
FIGURE 2
Variations at the chromophore level as found in different pyoverdine-related compounds.
10.1128/9781555816544/fig29-2_thmb.gif
10.1128/9781555816544/fig29-2.gif
FIGURE 2
Variations at the chromophore level as found in different pyoverdine-related compounds.
Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
/content/10.1128/9781555816544.chap29.fig29-3
FIGURE 3
IEF patterns of pyoverdines produced by Pseudomonas sp. strain LBSA1 (lane 1), P. fluorescens CIP 59.27 (lane 2), P. fluorescens ATCC 13525 (lane 3), P. fluorescens CIP 73.25 (lane 4), P. chlororaphis CIP 75.23 (lane 5), P. chlororaphis CIP 103295 (lane 6), P. fluorescens ATCC 17559 (lane 7), P. fluorescens CIP 104377 (lane 8), P. tolaasii CIP 106735 (lane 9), and the internal pHi standard (lane 10). A convenient device for electrofocusing is the mini-IEF gel apparatus from Bio-Rad. Following the manufacturer's recommendations, 5% polyacrylamide gels (10 by 6.5 cm; 0.4 mm thick) containing commercially available ampholines (the large pH 3 to 10 range is the most useful) are freshly cast (within 1 h), loaded with 10 to 20 samples (usually 1 _l of a 20 fold-concentrated CAA culture supernatant or 1 μl of a 5 mM purified siderophore aqueous solution), and electrophoresed for 15 min at 100 V, 15 min at 200 V, and 1 h at 450 V. The bright fluorescent pyoverdine bands are detected during exposure of the samples to UV light (350 nm).
10.1128/9781555816544/fig29-3_thmb.gif
10.1128/9781555816544/fig29-3.gif
FIGURE 3
IEF patterns of pyoverdines produced by Pseudomonas sp. strain LBSA1 (lane 1), P. fluorescens CIP 59.27 (lane 2), P. fluorescens ATCC 13525 (lane 3), P. fluorescens CIP 73.25 (lane 4), P. chlororaphis CIP 75.23 (lane 5), P. chlororaphis CIP 103295 (lane 6), P. fluorescens ATCC 17559 (lane 7), P. fluorescens CIP 104377 (lane 8), P. tolaasii CIP 106735 (lane 9), and the internal pHi standard (lane 10). A convenient device for electrofocusing is the mini-IEF gel apparatus from Bio-Rad. Following the manufacturer's recommendations, 5% polyacrylamide gels (10 by 6.5 cm; 0.4 mm thick) containing commercially available ampholines (the large pH 3 to 10 range is the most useful) are freshly cast (within 1 h), loaded with 10 to 20 samples (usually 1 _l of a 20 fold-concentrated CAA culture supernatant or 1 μl of a 5 mM purified siderophore aqueous solution), and electrophoresed for 15 min at 100 V, 15 min at 200 V, and 1 h at 450 V. The bright fluorescent pyoverdine bands are detected during exposure of the samples to UV light (350 nm).
Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
/content/10.1128/9781555816544.chap29.fig29-4
FIGURE 4
Heterologous pyoverdine-mediated 59Fe incorporation in P. costantinii CFBP 5705. Values on the ordinate correspond to the percentage of 59Fe incorporation after 20 min of incubation, compared to the homologous system. Numbers 1 to 35 on the abscissa correspond to the different pyoverdines tested, originating from the following bacterial strains: 1, Pseudomonas sp. strain E8; 2, P. syringae ATCC 19310; 3, P. fluorescens 9AW; 4, P. putida ATCC 12633; 5, P. fluorescens 51W; 6, P. aeruginosa, Pa6; 7, P. fluorescens CCM 2798; 8, P. fluorescens CHA0; 9, P. tolaasii LMG 2342; 10, P. aeruginosa ATCC 27853; 11, P. fluorescens ii; 12, P. fluorescens SB8.3; 13, P. fluorescens ATCC 17400; 14, P. fluorescens 1.3; 15, Pseudomonas sp. strain 267; 16, P. fluorescens ATCC 13525; 17, P. aeruginosa ATCC 15692; 18, P. fluorescens 18.1; 19, P. fluorescens 12; 20, P. fluorescens CFBP 2392; 21, P. putida CFBP 2461; 22, Pseudomonas sp. strain ATCC 15915; 23, P. monteilii CFML 90–54; 24, P. mosselii CFML 90–77; 25, P. rhodesiae CFML 92–104; 26, P. putida CFML 90–33; 27, Pseudomonas sp. strain CFML 90–40; 28, Pseudomonas sp. strain CFML 90–42; 29, Pseudomonas sp. strain CFML 90–51; 30, Pseudomonas sp. strain CFML 90.52; 31, Pseudomonas sp. strain 7SR1; 32, Pseudomonas sp. strain 2908; 33, Pseudomonas sp. strain A214; 34, P. aureofaciens; 35, P. costantinii CFBP 5705.
10.1128/9781555816544/fig29-4_thmb.gif
10.1128/9781555816544/fig29-4.gif
FIGURE 4
Heterologous pyoverdine-mediated 59Fe incorporation in P. costantinii CFBP 5705. Values on the ordinate correspond to the percentage of 59Fe incorporation after 20 min of incubation, compared to the homologous system. Numbers 1 to 35 on the abscissa correspond to the different pyoverdines tested, originating from the following bacterial strains: 1, Pseudomonas sp. strain E8; 2, P. syringae ATCC 19310; 3, P. fluorescens 9AW; 4, P. putida ATCC 12633; 5, P. fluorescens 51W; 6, P. aeruginosa, Pa6; 7, P. fluorescens CCM 2798; 8, P. fluorescens CHA0; 9, P. tolaasii LMG 2342; 10, P. aeruginosa ATCC 27853; 11, P. fluorescens ii; 12, P. fluorescens SB8.3; 13, P. fluorescens ATCC 17400; 14, P. fluorescens 1.3; 15, Pseudomonas sp. strain 267; 16, P. fluorescens ATCC 13525; 17, P. aeruginosa ATCC 15692; 18, P. fluorescens 18.1; 19, P. fluorescens 12; 20, P. fluorescens CFBP 2392; 21, P. putida CFBP 2461; 22, Pseudomonas sp. strain ATCC 15915; 23, P. monteilii CFML 90–54; 24, P. mosselii CFML 90–77; 25, P. rhodesiae CFML 92–104; 26, P. putida CFML 90–33; 27, Pseudomonas sp. strain CFML 90–40; 28, Pseudomonas sp. strain CFML 90–42; 29, Pseudomonas sp. strain CFML 90–51; 30, Pseudomonas sp. strain CFML 90.52; 31, Pseudomonas sp. strain 7SR1; 32, Pseudomonas sp. strain 2908; 33, Pseudomonas sp. strain A214; 34, P. aureofaciens; 35, P. costantinii CFBP 5705.
Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
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Tables
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy
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Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
/content/10.1128/9781555816544.chap29.tab29-1
TABLE 1
Possible side chains of pyoverdine isoforms
10.1128/9781555816544/tab29-1_thmb.gif
10.1128/9781555816544/tab29-1.gif
TABLE 1
Possible side chains of pyoverdine isoforms
Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
/content/10.1128/9781555816544.chap29.tab29-2
TABLE 2
Amino acid composition of the peptidic part of 44 pyoverdines with fully determined structures a
10.1128/9781555816544/tab29-2_thmb.gif
10.1128/9781555816544/tab29-2.gif
TABLE 2
Amino acid composition of the peptidic part of 44 pyoverdines with fully determined structures a
Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
/content/10.1128/9781555816544.chap29.tab29-3
TABLE 3
Examples of structurally related pyoverdines demonstrating cross-reactivity in pyoverdine-mediated iron uptake a
10.1128/9781555816544/tab29-3_thmb.gif
10.1128/9781555816544/tab29-3.gif
TABLE 3
Examples of structurally related pyoverdines demonstrating cross-reactivity in pyoverdine-mediated iron uptake a
Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
/content/10.1128/9781555816544.chap29.tab29-4
TABLE 4
Correlation between siderotype and species
10.1128/9781555816544/tab29-4_thmb.gif
10.1128/9781555816544/tab29-4.gif
TABLE 4
Correlation between siderotype and species
Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29
/content/10.1128/9781555816544.chap29.tab29-5
TABLE 5
The most common siderotypes and their corresponding siderovars found in 28 bacterial collections of diverse environmental origins representing 1,340 fluorescent Pseudomonas isolates a
10.1128/9781555816544/tab29-5_thmb.gif
10.1128/9781555816544/tab29-5.gif
TABLE 5
The most common siderotypes and their corresponding siderovars found in 28 bacterial collections of diverse environmental origins representing 1,340 fluorescent Pseudomonas isolates a
Citation:
Meyer J, Geoffroy V.
2004.
Environmental Fluorescent Pseudomonas and Pyoverdine Diversity: How Siderophores Could Help Microbiologists in Bacterial Identification and Taxonomy,
p 451-468.
In
Crosa J, Mey A, Payne S,
Iron Transport in Bacteria.
ASM Press, Washington, DC.
doi: 10.1128/9781555816544.ch29