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Applied and Environmental Microbiology, January 2004, p. 324-331, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.324-331.2004

Differential Stress-Induced Regulation of Two Quinone Reductases in the Brown Rot Basidiomycete Gloeophyllum trabeum

Roni Cohen,¹ Melissa R. Suzuki,² and Kenneth E. Hammel^1,2*

Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706,¹ Institute for Microbial and Biochemical Technology, USDA Forest Products Laboratory, Madison, Wisconsin 53726²

Received 10 July 2003/ Accepted 1 October 2003

Quinone reductases (QRDs) have two important functions in the basidiomycete Gloeophyllum trabeum, which causes brown rot of wood. First, a QRD is required to generate biodegradative hydroxyl radicals via redox cycling between two G. trabeum extracellular metabolites, 2,5-dimethoxyhydroquinone (2,5-DMHQ) and 2,5-dimethoxy-1,4-benzoquinone (2,5-DMBQ). Second, because 2,5-DMBQ is cytotoxic and 2,5-DMHQ is not, a QRD is needed to maintain the intracellular pool of these metabolites in the reduced form. Given their importance in G. trabeum metabolism, QRDs could prove useful targets for new wood preservatives. We have identified two G. trabeum genes, each existing in two closely related, perhaps allelic variants, that encode QRDs in the flavodoxin family. Past work with QRD1 and heterologous expression of QRD2 in this study confirmed that both genes encode NADH-dependent, flavin-containing QRDs. Real-time reverse transcription PCR analyses of liquid- and wood-grown cultures showed that qrd1 expression was maximal during secondary metabolism, coincided with the production of 2,5-DMBQ, and was moderately up-regulated by chemical stressors such as quinones. By contrast, qrd2 expression was maximal during fungal growth when 2,5-DMBQ levels were low, yet was markedly up-regulated by chemical stress or heat shock. The total QRD activity in lysates of G. trabeum mycelium was significantly enhanced by induction beforehand with a cytotoxic quinone. The promoter of qrd2 contains likely antioxidant, xenobiotic, and heat shock elements, absent in qrd1, that probably explain the greater response of qrd2 transcription to stress. We conclude from these results that QRD1 is the enzyme G. trabeum routinely uses to detoxify quinones during incipient wood decay and that it could also drive the biodegradative quinone redox cycle. However, QRD2 assumes a more important role when the mycelium is stressed.

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* Corresponding author. Mailing address: Institute for Microbial and Biochemical Technology, USDA Forest Products Laboratory, One Gifford Pinchot Dr., Madison, WI 53726. Phone: (608) 231-9528. Fax: (608) 231-9262. E-mail: kehammel{at}wisc.edu.

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Applied and Environmental Microbiology, January 2004, p. 324-331, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.324-331.2004

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