Abstract
ABSTRACTThe genus Malassezia includes 14 species that are found on the skin of humans and animals and are associated with a number of diseases. Recent genome sequencing projects have defined the gene content of all 14 species; however, to date, genetic manipulation has not been possible for any species within this genus. Here, we develop and then optimize molecular tools for the transformation of Malassezia furfur and Malassezia sympodialis using Agrobacterium tumefaciens delivery of transfer DNA (T-DNA) molecules. These T-DNAs can insert randomly into the genome. In the case of M. furfur, targeted gene replacements were also achieved via homologous recombination, enabling deletion of the ADE2 gene for purine biosynthesis and of the LAC2 gene predicted to be involved in melanin biosynthesis. Hence, the introduction of exogenous DNA and direct gene manipulation are feasible in Malassezia species.
Highlights
The genus Malassezia includes 14 species that are found on the skin of humans and animals and are associated with a number of diseases
The evolutionary trajectory toward becoming pathogenic must have differed, because Malassezia is more closely related to the smut plant pathogens like Ustilago maydis, while C. neoformans is related to fungal saprophytes [11]
At 37°C, M. sympodialis ATCC 42132 grew as well as the M. furfur strains in all medium types, while CBS 7222 showed no growth on media yeast extract-peptonedextrose (mYPD) and limited growth on the three remaining media, and strain ATCC 44340 was unable to grow on any medium tested at 37°C (Fig. 1)
Summary
The genus Malassezia includes 14 species that are found on the skin of humans and animals and are associated with a number of diseases. This research opens a new direction in terms of being able to analyze gene functions in this little understood genus These tools will contribute to define the mechanisms that lead to the commensalism and pathogenicity in this group of obligate fungi that are predominant on the skin of mammals. Wu and colleagues sequenced, assembled, and annotated genomes for all 14 known Malassezia species, including multiple strains of the most widely studied species (M. furfur, M. globosa, M. sympodialis, and M. restricta), for a total. Exceptions within the genus are represented by a subset of strains of M. furfur that have double the size of other M. furfur genomes, indicating genome duplication, hybridization events, or diploidy [9] Despite these extensive genome sequence resources, further work on Malassezia species is hampered by the current inability to modify their genes
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