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Genetics, Vol. 167, 619-631, June 2004, Copyright © 2004
doi:10.1534/genetics.103.023408
A Genetic Linkage Map of Cryptococcus neoformans variety neoformans Serotype D (Filobasidiella neoformans)
Robert E. Marra*,
,1,
Johnny C. Huang*,
Eula Fung
,
Kirsten Nielsen*,
Joseph Heitman*,
,
Rytas Vilgalys*, and
Thomas G. Mitchell*
* Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina 27710
Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina 27710
Department of Biology, Duke University, Durham, North Carolina 27710
Stanford Genome Technology Center, Palo Alto, California 94304
1 Corresponding author: Department of Plant Pathology and Ecology, Connecticut Agricultural Experiment Station, PO Box 1106, New Haven, CT 06504.
E-mail: robert.marra{at}po.state.ct.us
To construct a genetic linkage map of the heterothallic yeast, Cryptococcus neoformans (Filobasidiella neoformans), we crossed two mating-compatible strains and analyzed 94 progeny for the segregation of 301 polymorphic markers, consisting of 228 restriction site polymorphisms, 63 microsatellites, two indels, and eight mating-type (MAT)-associated markers. All but six markers showed no significant (P < 0.05) segregation distortion. At a minimum LOD score of 6.0 and a maximum recombination frequency of 0.30, 20 linkage groups were resolved, resulting in a map length of 
1500 cM. Average marker density is 5.4 cM (range 1–28.7 cM). Hybridization of selected markers to blots of electrophoretic karyotypes unambiguously assigned all linkage groups to chromosomes and led us to conclude that the C. neoformans genome is 20.2 Mb, comprising 14 chromosomes ranging in size from 0.8 to 2.3 Mb, with a ratio of 13.2 kb/cM averaged across the genome. However, only 2 of 12 ungrouped markers hybridized to chromosome 10. The hybridizations revealed at least one possible reciprocal translocation involving chromosomes 8, 9, and 12. This map has been critical to genome sequence assembly and will be essential for future studies of quantitative trait inheritance.
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