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Detection of antibiotic-related genes from bacterial biocontrol agents with polymerase chain reaction

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Pseudomonas chlororaphis PA23, Pseudomonas spp. strain DF41, and Bacillus amyloliquefaciens BS6 consistently inhibit infection of canola petals by Sclerotinia sclerotiorum in both greenhouse and field experiments. Bacillus thuringiensis BS8, Bacillus cereus L, and Bacillus mycoides S have shown significant inhibition against S. sclerotiorum on plate assays. The presence of antibiotic biosynthetic or self-resistance genes in these strains was investigated with polymerase chain reaction and, in one case, Southern blotting. Thirty primers were used to amplify (i) antibiotic biosythetic genes encoding phenazine-1-carboxylic acid, 2,4-diacetylphloroglucinol, pyoluteorin, and pyrrolnitrin, and (ii) the zwittermicin A self-resistance gene. Our findings revealed that the fungal antagonist P. chlororaphis PA23 contains biosynthetic genes for phenazine-1-carboxylic acid and pyrrolnitrin. Moreover, production of these compounds was confirmed by high performance liquid chromatography. Pseudomonas spp. DF41 and B. amyloliquefaciens BS6 do not appear to harbour genes for any of the antibiotics tested. Bacillus thuringiensis BS8, B. cereus L, and B. mycoides S contain the zwittermicin A self-resistance gene. This is the first report of zmaR in B. mycoides.Key words: Pseudomonas, Bacillus, biocontrol, antibiotic genes.

La souche PA23 de Pseudomonas chlororaphis, la souche DF41 de Pseudomonas spp. et la souche BS6 de Bacillus amyloliquefaciens ont régulièrement inhibé l'infection de pétales de canola par Sclerotinia sclerotiorum dans des expériences en serre et dans le champ. Bacillus thuringiensis BS8, Bacillus cereus L et Bacillus mycoides S ont démontré une inhibition significative de S. sclerotiorum dans des tests d'ensemencement. La présence de gènes de la biosynthèse d'antibiotiques ou d'auto-résistance chez ces souches fut évaluée à l'aide de la réaction de la polymérase en chaîne et dans un cas, de l'hybridation de type Southern. Trente amorces ont été utilisées pour amplifier (i) les gènes de la biosynthèse d'antibiotiques codant l'acide phenazine-1-carboxylique, le 2,4-diacetylphloroglucinol, la pyoluteorine, la pyrrolnitrine et (ii) le gène d'auto-résistance à la zwittermicine A. Nos découvertes révèlent que l'antagoniste fongique P. chlororaphis PA23 contient des gènes de la biosynthèse du l'acide phenazine-1-carboxylique et de la pyrrolnitrine. De plus, la production de ces composés fut confirmée par chromatographie liquide à haute performance. Pseudomonas spp. DF41 et B. amyloliquefaciens BS6 n'ont pas semblé renfermer de gènes d'aucun des antibiotiques analysés. Bacillus thuringiensis BS8, B. cereus L mycoides S contiennent le gène de l'auto-résistance à la zwittermicine A. Il s'agit de la première mention de zmaR chez B. mycoides.Mots clés : Pseudomonas, Bacillus, biocontrôle, gènes d'antibiotiques.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: May 1, 2006

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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