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Applied and Environmental Microbiology, October 2008, p. 6248-6253, Vol. 74, No. 20
0099-2240/08/$08.00+0     doi:10.1128/AEM.00970-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Formulation of a Defined V8 Medium for Induction of Sexual Development of Cryptococcus neoformans{triangledown}

Cory R. Kent,1,{dagger} Patricia Ortiz-Bermúdez,1,{dagger},{ddagger} Steven S. Giles,1 and Christina M. Hull1,2*

Department of Biomolecular Chemistry,1 Department of Medical Microbiology and Immunology, University of Wisconsin, School of Medicine and Public Health, Madison, Wisconsin 537062

Received 29 April 2008/ Accepted 24 August 2008

For over 3 decades, sexual development in the human fungal pathogen Cryptococcus neoformans and other fungi has been initiated by growing compatible mating partners on V8 juice medium. Although this medium is an efficient inducer of sexual development, the mechanism by which it promotes the process is unknown. To understand how V8 juice medium induces sexual development, we attempted to purify inducing factors from V8 juice, and we carried out a complete compositional analysis of V8 juice. We discovered that no single factor is responsible for the effects of V8 juice medium. Rather, the unique composition of V8 juice medium provides the proper nutrient composition for inducing and sustaining complete sexual development. Utilizing these findings, we developed a defined V8 (DV8) medium that mimics V8 juice medium in sexual development assays. Then, using DV8 as a tool, we explored the roles that specific molecules play in enhancing sexual development. Surprisingly, we discovered that copper is a key factor, leading to an upregulation of the mating pheromone genes MFa and MF{alpha}, both required for the initial steps in sexual development. The utilization of DV8 to investigate the effects of copper on sexual development presented here is an example of how defining the conditions that induce sexual development will advance the study of C. neoformans.

* Corresponding author. Mailing address: 587 Medical Science Center, 1300 University Avenue, Madison, WI 53706. Phone: (608) 265-5441. Fax: (608) 262-5253. E-mail: cmhull{at}wisc.edu

{triangledown} Published ahead of print on 5 September 2008.

{dagger} These authors contributed equally to this work.

{ddagger} Present address: Department of Chemical Engineering, University of Puerto Rico—Mayagüez, Mayagüez, PR 00680.

Applied and Environmental Microbiology, October 2008, p. 6248-6253, Vol. 74, No. 20
0099-2240/08/$08.00+0     doi:10.1128/AEM.00970-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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