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Applied and Environmental Microbiology, October 2002, p. 4996-5004, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.4996-5004.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Bcmfs1, a Novel Major Facilitator Superfamily Transporter from Botrytis cinerea, Provides Tolerance towards the Natural Toxic Compounds Camptothecin and Cercosporin and towards Fungicides

Keisuke Hayashi,{dagger} Henk-jan Schoonbeek,{ddagger} and Maarten A. De Waard*

Laboratory of Phytopathology, Wageningen University, 6700 EE Wageningen, The Netherlands

Received 28 March 2002/ Accepted 19 July 2002

Bcmfs1, a novel major facilitator superfamily gene from Botrytis cinerea, was cloned, and replacement and overexpression mutants were constructed to study its function. Replacement mutants showed increased sensitivity to the natural toxic compounds camptothecin and cercosporin, produced by the plant Camptotheca acuminata and the plant pathogenic fungus Cercospora kikuchii, respectively. Overexpression mutants displayed decreased sensitivity to these compounds and to structurally unrelated fungicides, such as sterol demethylation inhibitors (DMIs). A double-replacement mutant of Bcmfs1 and the ATP-binding cassette (ABC) transporter gene BcatrD was more sensitive to DMI fungicides than a single-replacement mutant of BcatrD, known to encode an important ABC transporter of DMIs. The sensitivity of the wild-type strain and mutants to DMI fungicides correlated with Bcmfs1 expression levels and with the initial accumulation of oxpoconazole by germlings of these isolates. The results indicate that Bcmfs1 is a major facilitator superfamily multidrug transporter involved in protection against natural toxins and fungicides and has a substrate specificity that overlaps with the ABC transporter BcatrD. Bcmfs1 may be involved in protection of B. cinerea against plant defense compounds during the pathogenic phase of growth on host plants and against fungitoxic antimicrobial metabolites during its saprophytic phase of growth.

* Corresponding author. Mailing address: Laboratory of Phytopathology, Wageningen University, P.O. Box 8025, 6700 EE Wageningen, The Netherlands. Phone: 31 317 483123. Fax: 31 317 483412. E-mail: maarten.dewaard{at}wur.nl.

{dagger} Present address: Ube Research Laboratory, Ube Industries, Ltd., 1978-5 Kogushi, Ube, Yamaguchi, Japan.

{ddagger} Present address: Department of Plant Biology, University of Fribourg, CH-1700 Fribourg, Switzerland.

Applied and Environmental Microbiology, October 2002, p. 4996-5004, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.4996-5004.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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