Phylogeny and intercontinental distribution of the pneumocandin-producing anamorphic fungus Glarea lozoyensis

Abstract

Glarea lozoyensis is an anamorphic ascomycete that produces pneumocandin B0, the starting molecule for the synthesis of the antifungal drug caspofungin (CANCIDAS™). Glarea lozoyensis was first isolated in 1985 from a water sample from Madrid, Spain. Until now, only the original strain was known, but we have discovered new strains from Argentina and the USA. Molecular phylogenetic reference to a 28S rDNA database of antibiotic-producing fungi quickly identified these strains as being conspecific with G. lozoyensis. Bayesian inference phylogeny of ITS, 28S rDNA and α-actin gene fragments revealed that G. lozoyensis is related to species of the genus Cyathicula (Helotiales). Glarea lozoyensis was not conspecific with any of the Cyathicula species sequenced, although it appears to share a common ancestor. Glarea lozoyensis and Cyathicula strains were fermented on nutritional microarrays in 96-well plates. Cyathicula extracts did not show antifungal activity and did not produce pneumocandins, whereas potent antifungal activity and pneumocandin A0 production were confirmed for the four G. lozoyensis isolates. Also, culture morphology differed, with G. lozoyensis strains producing a dark brown, profusely sporulating mycelium with pigmented multicellular conidia accumulating in conidial masses, while all Cyathicula species tested in culture formed hyaline to light brown mycelia and lacked conidia. The chemistry and taxonomic distribution of the echinocandin class of antifungals is comprehensively reviewed.