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Title: DETECTION OF ANTIBIOTIC-RELATED GENES FROM BACTERIAL BIOCONTROL AGENTS WITH POLYMERASE CHAIN REACTION

Author
item ZHANG, Y - UNIV. OF MANITOBA
item FERNANDO, W. G. - UNIV. OF MANITOBA
item DE KIEVIT, T. - UNIV. OF MANITOBA
item BERRY, C - UNIV. OF MANITOBA
item DAAYF, F - UNIV. OF MANITOBA
item Paulitz, Timothy

Submitted to: Canadian Journal of Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/5/2005
Publication Date: 4/20/2006
Citation: Zhang, Y., Fernando, W.D., De Kievit, T.R., Berry, C., Daayf, F., Paulitz, T.C. 2006. Detection of antibiotic-related genes from bacterial biocontrol agents with polymerase chain reaction. Canadian Journal of Microbiology. Vol 52:476-481.

Interpretive Summary: Bacterial biocontrol agents were tested for the control of white rot of canola caused by Sclerotinia sclerotiorum. These isolates were tested with PCR probes for the presence of genes coding for known antifungal compounds. Pseudomonas. chlororaphis PA23 contains biosynthetic genes for phenazine-1-carboxylic acid and pyrrolnitrin., and this was confirmed by HPLC. 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

Technical Abstract: 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