Genetics. Published Articles Ahead of Print: December 1, 2005, Copyright © 2005
doi:10.1534/genetics.105.046672


A more recent version of this article appeared on April 1, 2006.

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Multilocus Sequence Typing Reveals Three Genetic Subpopulations of Cryptococcus neoformans var. grubii (serotype A), including a Unique Population in Botswana

Anastasia P. Litvintseva 1*, Rameshwari Thakur 2, Rytas Vilgalys 3 and Thomas G. Mitchell 1

1 Duke University Medical Center
2 National Health Laboratory, Ministry of Health, Gaborone, Botswana
3 Duke University

* To whom correspondence should be addressed. E-mail: litvi001{at}mc.duke.edu.

Submitted on June 8, 2005
Revised on August 15, 2005
Accepted on 28 October 2005


Abstract

We applied multilocus sequence typing (MLST) to investigate the population structure and mode of reproduction of Cryptococcus neoformans var. grubii (serotype A). This MLST system utilizes 12 unlinked polymorphic loci, which are dispersed on nine different chromosomes, and allows the unambiguous identification of closely related strains of serotype A. We compared MLST analyses with the conventional genotyping method of detecting amplified fragment length polymorphisms (AFLP), and there was excellent correlation between the MLST and AFLP results. However, MLST differentiated a larger number of strains. We analyzed a global collection of isolates of serotype A using both methods, and the results identified at least three genetically distinct subpopulations, designated Groups VNI, VNII, and VNB. Groups VNI and VNII are widespread, dominated by isolates with the MAT{alpha} mating type, and predominantly clonal. Conversely, isolates of group VNB are unique to Botswana, include a significant proportion of fertile strains with the MAT{alpha} mating type, and manifest compelling evidence of recombination. We have AFLP genotyped more than 1,000 strains of serotype A from different parts of the world, including isolates from several African countries, and to date, haploid serotype A isolates of Group VNB have only been found in Botswana.

Key Words: AFLP, Cryptococcus neoformans, MLST, population structure, recombination




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