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Molecular and Cellular Biology, January 2000, p. 329-339, Vol. 20, No. 1
0270-7306/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of a Ustilago maydis Gene Specifically Induced during the Biotrophic Phase: Evidence for Negative as Well as Positive Regulation

Christoph W. Basse, Stefan Stumpferl,dagger and Regine Kahmann*

Institut für Genetik und Mikrobiologie der Universität München, D-80638 Munich, Germany

Received 27 July 1999/Returned for modification 3 September 1999/Accepted 4 October 1999

The phytopathogenic basidiomycete Ustilago maydis requires its host plant, maize, for completion of its sexual cycle. To investigate the molecular events during infection, we used differential display to identify plant-induced U. maydis genes. We describe the U. maydis gene mig1 (for "maize-induced gene"), which is not expressed during yeast-like growth of the fungus, is weakly expressed during filamentous growth in axenic culture, but is extensively upregulated during plant infection. mig1 encodes a small, highly charged protein of unknown function which contains a functional N-terminal secretion sequence and is not essential for pathogenic development. Adjacent to mig1 is a second gene (mdu1) related to mig1, which appears to result from a gene duplication. mig1 gene expression during the infection cycle was assessed by fusing the promoter to eGFP. Expression of mig1 was absent in hyphae growing on the leaf surface but was detected after penetration and remained high during subsequent proliferation of the fungus until teliospore formation. Successive deletions as well as certain internal deletions in the mig1 promoter conferred elevated levels of reporter gene expression during growth in axenic culture, indicative of negative regulation. During fungal growth in planta, sequence elements between positions -148 and -519 in the mig1 promoter were specifically required for high levels of induction, illustrating additional positive control. We discuss the potential applications of mig1 for the identification of inducing compounds and the respective regulatory genes.


* Corresponding author. Mailing address: Institut für Genetik und Mikrobiologie, Maria-Ward-Str.1a, D-80638 München, Germany. Phone: 49-89-21806150. Fax: 49-89-1785633. E-mail: R.Kahmann{at}lrz.uni-muenchen.de.

dagger Present address: Biozentrum Niederursel, Goethe Universität, D-60439 Frankfurt, Germany.


Molecular and Cellular Biology, January 2000, p. 329-339, Vol. 20, No. 1
0270-7306/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



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