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Microbiology 149 (2003), 131-142; DOI  10.1099/mic.0.25921-0
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Microbiology 149 (2003), 131-142; DOI  10.1099/mic.0.25921-0
© 2003 Society for General Microbiology

Molecular and genetic characterization of a serotype A MATa Cryptococcus neoformans isolate

S. M. Keller1, M. A. Viviani2, M. C. Esposto2, M. Cogliati2 and B. L. Wickes1

1 Department of Microbiology, Mail Code 7758, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA
2 Laboratorio di Micologia Medica, Instituto di Igiene e Medicina Preventiva, Universita degli Studi di Milano, IRCCS Ospedale Maggiore, Milano, Italy

Correspondence
B. L. Wickes
wickes{at}uthscsa.edu

Cryptococcus neoformans comprises two varieties (neoformans and gattii) and four serotypes (A, B, C and D). Fertile isolates of both mating types have been identified in serotypes B, C and D; however, a fertile serotype A MATa strain has not been confirmed, although serotype A MAT{alpha} strains will mate with serotype D MATa strains. Preliminary analysis of a recent Italian environmental isolate (IUM 96-2828) suggested that this strain was haploid, serotype A and MATa. In this study, IUM 96-2828 has been characterized in detail. A mating reaction between IUM 96-2828 and H99 (serotype A MAT{alpha}) produced abundant spores with an equal distribution of MATa and MAT{alpha} progeny, all of which were serotype A. Karyotypic analysis of F1 spores revealed evidence of recombination, confirming that IUM 96-2828 was fertile. The MATa pheromone gene from IUM 96-2828 was sequenced and found to be most closely related to the serotype D MATa pheromone gene. Phylogenetic comparisons of other genes not linked to mating type also suggested IUM 96-2828 was most closely related to serotype A strains. Biochemical analysis showed that the carbon assimilation profiles of H99 and IUM 96-2828 were identical for 97 % (30/31) of the substrates while isozyme analysis showed 89 % (17/19) identity. Assays of major virulence factors found no difference between H99 and IUM 96-2828. Virulence studies using the mouse model demonstrated that IUM 96-2828 was virulent for mice, although it was less virulent than H99. These data strongly suggest that IUM 96-2828 is a true haploid serotype A MATa isolate that is fertile.

The GenBank accession number for the MFa1A pheromone sequence reported in this paper is AY129299.




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