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Eukaryotic Cell, January 2006, p. 155-166, Vol. 5, No. 1
1535-9778/06/$08.00+0     doi:10.1128/EC.5.1.155-166.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Cdc42 Is Required for Proper Growth and Development in the Fungal Pathogen Colletotrichum trifolii

Changbin Chen, Young-sil Ha, Ji-young Min, Stephen D. Memmott, and Martin B. Dickman*

Department of Plant Pathology, University of Nebraska—Lincoln, Lincoln, Nebraska 68583-0722

Received 3 June 2005/ Accepted 22 October 2005

Cdc42 is a highly conserved small GTP-binding protein that is involved in regulating morphogenesis in eukaryotes. In this study, we isolated and characterized a highly conserved Cdc42 gene from Colletotrichum trifolii (CtCdc42), a fungal pathogen of alfalfa. CtCdc42 is, at least in part, functionally equivalent to Saccharomyces cerevisiae Cdc42p, since it restores the temperature-sensitive phenotype of a yeast Cdc42p mutant. Inhibition of CtCdc42 by expression of an antisense CtCdc42 or a dominant negative form of CtCdc42 (DN Cdc42) resulted in appressorium differentiation under noninductive conditions, suggesting that CtCdc42 negatively regulates pathogenic development in this fungus. We also examined the possible linkage between CtCdc42 and Ras signaling. Expression of a dominant active Cdc42 (DA Cdc42) in C. trifolii leads to aberrant hyphal growth under nutrient-limiting conditions. This phenotype was similar to that of our previously reported dominant active Ras (DA Ras) mutant. Also consistent with our observations of the DA Ras mutant, high levels of reactive oxygen species (ROS) were observed in the DA Cdc42 mutant, and proline restored the wild-type phenotype. Moreover, overexpression of DN Cdc42 resulted in a significant decrease in spore germination, virtually no hyphal branching, and earlier sporulation, again similar to what we observed in a dominant negative Ras (DN Ras) mutant strain. Interestingly, coexpression of DA Cdc42 with DN Ras resulted in germination rates close to wild-type levels, while coexpression of DN Cdc42 with the DA Ras mutant restored the wild-type phenotype. These data suggest that CtCdc42 is positioned as a downstream effector of CtRas to regulate spore germination and pathogenic development.


* Corresponding author. Mailing address: Department of Plant Pathology, 406 Plant Sciences Hall, University of Nebraska—Lincoln, Lincoln, NE 68583-0722. Phone: (402) 472-2849. Fax: (402) 472-2853. E-mail: mdickman{at}unlnotes.unl.edu.


Eukaryotic Cell, January 2006, p. 155-166, Vol. 5, No. 1
1535-9778/06/$08.00+0     doi:10.1128/EC.5.1.155-166.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.




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