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J Bacteriol, April 1998, p. 2021-2026, Vol. 180, No. 8
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Iron Uptake in Ustilago maydis: Tracking the Iron Path

Orly Ardon,1 Raphael Nudelman,2 Catherine Caris,2 Jacqueline Libman,2,§ Abraham Shanzer,2 Yona Chen,3 and Yitzhak Hadar1,*

Department of Plant Pathology and Microbiology and The Otto Warburg Center for Agricultural Biotechnology1 and Department of Soil and Water Sciences,3 Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, and Department of Organic Chemistry, The Weizmann Institute of Science,2 Rehovot 76100, Israel

Received 2 January 1998/Accepted 6 February 1998

In this study, we monitored and compared the uptake of iron in the fungus Ustilago maydis by using biomimetic siderophore analogs of ferrichrome, the fungal native siderophore, and ferrioxamine B (FOB), a xenosiderophore. Ferrichrome-iron was taken up at a higher rate than FOB-iron. Unlike ferrichrome-mediated uptake, FOB-mediated iron transport involved an extracellular reduction mechanism. By using fluorescently labeled siderophore analogs, we monitored the time course, as well as the localization, of iron uptake processes within the fungal cells. A fluorescently labeled ferrichrome analog, B9-lissamine rhodamine B, which does not exhibit fluorescence quenching upon iron binding, was used to monitor the entry of the compounds into the fungal cells. The fluorescence was found intracellularly 4 h after the application and later was found concentrated in two to three vesicles within each cell. The fluorescence of the fluorescently labeled FOB analog CAT18, which is quenched by iron, was visualized around the cell membrane after 4 h of incubation with the ferrated (nonfluorescent) compounds. This fluorescence intensity increased with time, demonstrating fungal iron uptake from the siderophores, which remained extracellular. We here introduce the use of fluorescent biomimetic siderophores as tools to directly track and discriminate between different pathways of iron uptake in cells.

* Corresponding author. Mailing address: Department of Plant Pathology and Microbiology and The Otto Warburg Center for Agricultural Biotechnology, The Hebrew University of Jerusalem, Faculty of Agricultural, Food and Environmental Quality Sciences, Rehovot 76100, Israel. Phone: 972 8 9481315. Fax: 972 8 9468785. E-mail: hadar{at}agri.huji.ac.il.

§ Jacqueline Libman passed away on March 30, 1997, while this work was in progress. This article is dedicated in her memory.




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