Cloning of genes involved in the biosynthesis of pseudobactin, a high-affinity iron transport agent of a plant growth-promoting Pseudomonas strain

Moores, Jane C.; Magazin, Marilyn; Ditta, Gary S.; Leong, John M.

doi:10.1128/jb.157.1.53-58.1984

Cited by 73 publications

(24 citation statements)

References 34 publications

(32 reference statements)

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“…This fits well with the situation in Pseudomonas sp. strain B10, where minimally 12 genes in four separate clusters are needed for the biosynthesis of pseudobactin (22).…”

Section: Discussionmentioning

confidence: 99%

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Isolation and analysis of genes involved in siderophore biosynthesis in plant-growth-stimulating Pseudomonas putida WCS358

Marugg¹,

Spanje²,

Hoekstra³

et al. 1985

J Bacteriol

115

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The plant-growth-stimulating Pseudomonas putida WCS358 was mutagenized with transposon TnS. The resulting mutant colony bank was screened for mutants defective in the biosynthesis of the fluorescent siderophore. A total of 28 mutants, divided into six different classes, were isolated that were nonfluorescent or defective in iron acquisition or both. These different types of mutants together with the probable overall structure of the siderophore, i.e., a small peptide chain attached to a fluorescing group, suggest a biosynthetic pathway in which the synthesis of the fluorescing group is preceded by the synthesis of the peptide part. A gene colony bank of P. putida WCS358 was constructed with the broad-host-range cosmid vector pLAFR1. This genomic library, established in Escherichia coli, was mobilized into the 28 individual mutants, screening for transconjugants restored in fluorescence or growth under iron-limiting conditions or both. A total of 13 cosmids were found to complement 13 distinct mutants. The complementation analysis revealed that at least five gene clusters, with a minimum of seven genes, are needed for siderophore biosynthesis. Some of these genes seem to be arranged in an operon-like structure. * Corresponding author. hexapeptide are involved in the iron(III) binding. The structures of pyoverdine, produced by P. fluorescens (24,25) andPseudomonas aeruginosa (32), and of pseudobactin 7SR1, the siderophore of the plant-deleterious Pseudomonas sp.strain 7SR1 (34), were found to be very similar to that of pseudobactin. These findings suggest a common structure for siderophores of various fluorescent Pseudomonas species, although it is stated that at least minor structural differences exist among the fluorescent pigments (20).

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“…This fits well with the situation in Pseudomonas sp. strain B10, where minimally 12 genes in four separate clusters are needed for the biosynthesis of pseudobactin (22).…”

Section: Discussionmentioning

confidence: 99%

“…Recently, complementation studies with nonfluorescent mutants, defective in siderophore biosynthesis, of Pseudomonas sp. strain B10 have indicated that a minimum of 12 genes arranged in four gene clusters is required for the biosynthesis of pseudobactin (22).…”

mentioning

confidence: 99%

Isolation and analysis of genes involved in siderophore biosynthesis in plant-growth-stimulating Pseudomonas putida WCS358

Marugg¹,

Spanje²,

Hoekstra³

et al. 1985

J Bacteriol

115

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“…All recombinant DNA procedures have been described elsewhere [17]. Cosmids carrying di¡erent Pseudomonas B10 genomic fragments [19] were transferred from E. coli HB101 to P. aeruginosa by triparental matings with pRK2013 [20]. Exconjugants were selected on SM9 and screened for £uorescent emission under UV light exposure.…”

Section: Manipulation Of Nucleic Acids and Genetic Techniquesmentioning

confidence: 99%

“…This observation indicates that PvdS is required for PpsbA transcription in the heterologous P. aeruginosa host, and suggests that a pvdS homolog is present in Pseudomonas B10. To search for this gene, eight cosmids encompassing the pseudobactin region of the Pseudomonas B10 genome [19] were individually screened for their ability to complement the pvdS mutation P. aeruginosa PAO OT11pvdS [8]. Cosmid pJLM4A, containing a DNA insert of 26.8 kb, was able to restore the £uorescent phenotype in PAO OT11pvdS.…”

Section: Identi¢cation Of the Psbs Genementioning

confidence: 99%

Transcriptional regulation of pseudobactin synthesis in the plant growth-promoting Pseudomonas B10

Leoni¹

2002

FEMS Microbiology Letters

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We have investigated the iron-dependent regulation of the psbA gene, encoding the enzyme L-ornithine N 5 -oxygenase in the rhizobacterium Pseudomonas B10. We have cloned and characterized a Pseudomonas B10 gene, designated psbS, required for psbA expression. PsbS is endowed with structural and functional features of extracytoplasmatic function (ECF) sigma factors, and is closely related to the iron starvation sigmas PvdS, PbrA, and PfrI, which mediate the iron-repressible expression of pseudobactin biosynthesis genes in different Pseudomonas species. Expression of psbA was found to be indirectly controlled by Fur, which abrogates psbS transcription in the presence of sufficient iron. ß

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“…Eight pLAFR1-derived cosmids from a genomic library of Pseudomonas strain B10 (25) were individually screened for the ability to complement the pvdA mutation in the nonfluorescent, L-Orn N 5 -hydroxylase-deficient Pseudomonas aeruginosa mutant PALS124 (pvdA). Only pJLM1, carrying a DNA insert of approximately 23 kb (25), was able to restore the fluorescent phenotype in PALS124 grown on KB agar plates (13). Subcloning in the shuttle vector pUCP18 and deletion analysis localized the complementing DNA region within the 2.7-kb PstI-SphI insert of pCA⌬Sh.…”

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confidence: 99%

Pseudobactin Biogenesis in the Plant Growth-Promoting Rhizobacterium Pseudomonas Strain B10: Identification and Functional Analysis of the l -Ornithine N ⁵ -Oxygenase ( psbA ) Gene

Ambrosi¹,

Leoni²,

Putignani³

et al. 2001

J Bacteriol

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Pseudobactin B10 , the fluorescent siderophore produced by the rhizobacterium Pseudomonas strain B10, contains the hydroxamate ligand D-N 5 -hydroxyornithine (D-N 5 -OH-Orn). We cloned the L-Orn N 5 -oxygenase (psbA) gene from a genomic library of Pseudomonas strain B10 and demonstrated that PsbA is involved in the conversion of L-Orn to its N 5 -OH derivative. PsbA shows significant similarity to microbial -amino acid hydroxylases containing flavin adenine dinucleotide and NADP cofactor-binding sites and the FATGY signature of the putative substrate recognition pocket. The psbA gene is monocistronic, and its transcription is negatively controlled by iron. A site-specific psbA mutant of Pseudomonas strain B10 was biochemically complemented with the precursor L-N 5 -OH-Orn, suggesting that L-Orn is hydroxylated before conversion to the D isomer. The L-Orn N 5 -hydroxylase-defective mutants of Pseudomonas strain B10 and Pseudomonas aeruginosa PAO1 were much less effective than the parental strains in suppressing the growth of the phytopathogen Erwinia carotovora in iron-poor medium. The extent of in vitro inhibition of E. carotovora was strictly iron dependent and directly correlated with the amount of released siderophores. These data strengthen the role of fluorescent siderophores in biocontrol of deleterious rhizomicroorganisms.

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Cloning of genes involved in the biosynthesis of pseudobactin, a high-affinity iron transport agent of a plant growth-promoting Pseudomonas strain

Cited by 73 publications

References 34 publications

Isolation and analysis of genes involved in siderophore biosynthesis in plant-growth-stimulating Pseudomonas putida WCS358

Isolation and analysis of genes involved in siderophore biosynthesis in plant-growth-stimulating Pseudomonas putida WCS358

Transcriptional regulation of pseudobactin synthesis in the plant growth-promoting Pseudomonas B10

Pseudobactin Biogenesis in the Plant Growth-Promoting Rhizobacterium Pseudomonas Strain B10: Identification and Functional Analysis of the l -Ornithine N ⁵ -Oxygenase ( psbA ) Gene

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Cloning of genes involved in the biosynthesis of pseudobactin, a high-affinity iron transport agent of a plant growth-promoting Pseudomonas strain

Cited by 73 publications

References 34 publications

Isolation and analysis of genes involved in siderophore biosynthesis in plant-growth-stimulating Pseudomonas putida WCS358

Isolation and analysis of genes involved in siderophore biosynthesis in plant-growth-stimulating Pseudomonas putida WCS358

Transcriptional regulation of pseudobactin synthesis in the plant growth-promoting Pseudomonas B10

Pseudobactin Biogenesis in the Plant Growth-Promoting Rhizobacterium Pseudomonas Strain B10: Identification and Functional Analysis of the l -Ornithine N 5 -Oxygenase ( psbA ) Gene

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Pseudobactin Biogenesis in the Plant Growth-Promoting Rhizobacterium Pseudomonas Strain B10: Identification and Functional Analysis of the l -Ornithine N ⁵ -Oxygenase ( psbA ) Gene