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J Biol Chem, Vol. 275, Issue 19, 14242-14247, May 12, 2000

Ornithine Decarboxylase of Stagonospora (Septoria) nodorum Is Required for Virulence toward Wheat*

Andy BaileyDagger , Elisabeth Mueller§, and Paul Bowyer

From the Institute of Arable Crop Research-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, Long Ashton, Bristol, BS41 9AF, United Kingdom

A knockout strain of Stagonospora (Septoria) nodorum lacking the single ornithine decarboxylase (ODC) allele has been created by targeted gene replacement. A central region of the S. nodorum ODC gene was isolated by polymerase chain reaction using degenerate oligonucleotides and used to probe a lambda  genomic library. The gene was sequenced and the encoded ODC protein sequence was shown to be similar to those from other fungi. The functionality of the S. nodorum ODC was confirmed by complementation of an Aspergillus nidulans mutant (puA) strain devoid of ODC activity, restoring growth in the absence of exogenous polyamines. Sporulation of the transformants was reduced suggesting abberant regulation of the S. nodorum gene in A. nidulans. Transformation-mediated gene replacement was used to create strains which were auxotrophic for putrescine and lack ODC coding sequences. Pathogenicity studies on these mutants showed that they are greatly reduced in virulence compared with non-disrupted transformants. This confirms that the strains carrying an ODC disruption cannot obtain sufficient polyamines from the host plant for normal growth and, thus, that fungal ODC may be a suitable target for chemical intervention.

* This work was supported by the Institute of Arable Crop Research-Long Ashton Research Station, supported by the Biotechnology and Biological Sciences Research Council (BBSRC).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Recipient of a BBSRC Biological Chemistry Initiative grant. Current address: Dept. of Biological Sciences, University of Bristol, Bristol, UK.

§ Supported by a BBSRC Cooperative Awards in Science and Engineering studentship award co-funded by Zeneca Agrochemicals.

To whom all correspondence should be addressed. Tel.: 1275-549297; Fax: 1275-394281; E-mail: bowyerp@bbsrc.ac.uk.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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