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Applied and Environmental Microbiology, May 2001, p. 2088-2094, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2088-2094.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Induction of Laccase Activity in Rhizoctonia solani by Antagonistic Pseudomonas fluorescens Strains and a Range of Chemical Treatments

Jonathan D. Crowe and Stefan Olsson*

Section of Genetics and Microbiology, Department of Ecology, The Royal Veterinary and Agricultural University, DK-1871 Frederiksberg, Copenhagen, Denmark

Received 11 December 2000/Accepted 26 February 2001

Fungi often produce the phenoloxidase enzyme laccase during interactions with other organisms, an observation relevant to the development of biocontrols. By incorporating the laccase substrate 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) into agar, we analyzed laccase induction in the plant-pathogenic fungus Rhizoctonia solani when paired against isolates of the soil bacterium Pseudomonas fluorescens. Substantial induction of R. solani laccase was seen only in pairings with strains of P. fluorescens known to produce antifungal metabolites. To study laccase induction further, a range of chemical treatments was applied to R. solani liquid cultures. p-Anisidine, copper(II), manganese(II), calcium ionophore A23187, lithium chloride, calcium chloride, cyclic AMP (cAMP), caffeine, amphotericin B, paraquat, ethanol, and isopropanol were all found to induce laccase; however, the P. fluorescens metabolite viscosinamide did not do so at the concentrations tested. The stress caused by these treatments was assessed by measuring changes in lipid peroxidation levels and dry weight. The results indicated that the laccase induction seen in pairing plate experiments was most likely due to calcium or heat shock signaling in response to the effects of bacterial metabolites, but that heavy metal and cAMP-driven laccase induction was involved in sclerotization.


* Corresponding author. Mailing address: The Royal Veterinary and Agricultural University, Department of Ecology, Section of Genetics and Microbiology, 40, Thorvaldsensvej, DK-1871 Frederiksberg, Copenhagen, Denmark. Phone: (45) 35282646. Fax: (45) 35282606. E-mail: stefan.olsson{at}ecol.kvl.dk.


Applied and Environmental Microbiology, May 2001, p. 2088-2094, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2088-2094.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.