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Flutolanil and carboxin resistance in Coprinus cinereus conferred by a mutation in the cytochrome b 560 subunit of succinate dehydrogenase complex (Complex II)

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Abstract

A gene that confers resistance to the systemic fungicide flutolanil was isolated from a mutant strain of the basidiomycete Coprinus cinereus. The flutolanil resistance gene was mapped to a chromosome of approximately 3.2 Mb, and a chromosome-specific cosmid library was constructed. Two cosmid clones that were able to transform a wild-type, flutolanil-sensitive, strain of C. cinereus to resistance were isolated from the library. Analysis of a subclone containing the resistance gene revealed the presence of the sdhC gene, which encodes the cytochrome b 560 subunit of the succinate dehydrogenase (SDH) complex (Complex II) in the mitochondrial membrane. Comparison between the sdhC gene of a wild-type strain and that of a mutant strain revealed a single point mutation, which results in the replacement of Asn by Lys at position 80. Measurements of succinate-cytochrome c reductase activity in the transformants with mutant sdhC gene(s) suggest that flutolanil resistance of the fungus is caused by a decrease in the affinity of the SDH complex for flutolanil. This sdhC mutation also conferred cross-resistance against another systemic fungicide, carboxin, an anilide that is structurally related to flutolanil. In other organisms carboxin resistance mutations have been found in the genes sdhB and sdhD, but this is the first demonstration that a mutation in sdhC can also confer resistance. The mutant gene cloned in this work can be utilized as a dominant selectable marker in gene manipulation experiments in C. cinereus.

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Acknowledgements

We are grateful to Dr. Takashi Kamada for helpful advice on constructing the chromosome-specific cosmid library, to Dr. Patricia J. Pukkila for kindly consenting to our use of the cosmid vector, and to Dr. Miriam E. Zolan for critical reading of the manuscript. This work was supported by a grant from the Ministry of Agriculture, Forestry, and Fisheries of Japan (Development of Innovative Plants and Animals using Transformation and Cloning, Project 2102)

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Authors and Affiliations

  1. Biological Function Division, National Food Research Institute, 305-8642, Tsukuba, Ibaraki, Japan

    Y. Ito, Y. Seshime & S. Oita

  2. Department of Biotechnology, Faculty of Bioresource Science, Akita Prefectural University, 010-0195, Akita, Japan

    H. Muraguchi & S. O. Yanagi

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  1. Y. Ito
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  2. H. Muraguchi
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  3. Y. Seshime
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  4. S. Oita
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  5. S. O. Yanagi
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Correspondence to Y. Ito.

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Communicated by E. Cerdá-Olmedo

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Ito, Y., Muraguchi, H., Seshime, Y. et al. Flutolanil and carboxin resistance in Coprinus cinereus conferred by a mutation in the cytochrome b 560 subunit of succinate dehydrogenase complex (Complex II). Mol Genet Genomics 272, 328–335 (2004). https://doi.org/10.1007/s00438-004-1060-2

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  • DOI: https://doi.org/10.1007/s00438-004-1060-2

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