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Trichosporon asahii  is a pathogenic fungus that causes severe, deep-seated fungal infections in neutropenic patients .  Elucidating the infection mechanisms of  T. asahii  based on genetic studies requires a specific gene-targeting system. Here, we established an efficient gene-targeting system in a highly pathogenic  T. asahii  strain identified using the silkworm infection model. By comparing the pathogenicity of  T. asahii  clinical isolates in a silkworm infection model,  T. asahii  MPU129 was identified as a highly pathogenic strain. Using an  Agrobacterium tumefaciens -mediated gene transfer system, we obtained a  T. asahii  MPU129 mutant lacking the  ku70  gene, which encodes the Ku70 protein involved in the non-homologous end-joining repair of DNA double-strand breaks. The  ku70  gene-deficient mutant showed higher gene-targeting efficiency than the wild-type strain for constructing a mutant lacking the  cnb1  gene, which encodes the beta-subunit of calcineurin. The  cnb1  gene-deficient mutant showed reduced pathogenicity against silkworms compared with the parental strain. These results suggest that an efficient gene-targeting system in a highly pathogenic  T. asahii  strain is a useful tool for elucidating the molecular mechanisms of  T. asahii  infection.

Development of an efficient gene-targeting system for elucidating infection mechanisms of the fungal pathogen Trichosporon asahii