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Background  Nodulisporic acids (NAs) are indole diterpene fungal metabolites exhibiting potent systemic efficacy against blood-feeding arthropods, e.g., bedbugs, fleas and ticks, via binding to arthropod specific glutamate-gated chloride channels. Intensive medicinal chemistry efforts employing a nodulisporic acid A template have led to the development of N- tert -butyl nodulisporamide as a product candidate for a once monthly treatment of fleas and ticks on companion animals. The source of the NAs is a monophyletic lineage of asexual endophytic fungal strains that is widely distributed in the tropics, tentatively identified as a  Nodulisporium  species and hypothesized to be the asexual state of a  Hypoxylon  species.    Methods and Results  Inferences from GenBank sequences indicated that multiple researchers have encountered similar  Nodulisporium  endophytes in tropical plants and in air samples. Ascomata-derived cultures from a wood-inhabiting fungus, from Martinique and closely resembling  Hypoxylon investiens,  belonged to the same monophyletic clade as the NAs-producing endophytes. The hypothesis that the Martinique  Hypoxylon  collections were the sexual state of the NAs-producing endophytes was tested by mass spectrometric analysis of NAs, multi-gene phylogenetic analysis, and phenotypic comparisons of the conidial states. We established that the Martinique  Hypoxylon  strains produced an ample spectrum of NAs and were conspecific with the pantropical  Nodulisporium  endophytes, yet were distinct from  H. investiens . A new species,  H. pulicicidum , is proposed to accommodate this widespread organism.    Conclusions and Significance  Knowledge of the life cycle of  H. pulicicidum  will facilitate an understanding of the role of insecticidal compounds produced by the fungus, the significance of its infections in living plants and how it colonizes dead wood. The case of  H. pulicicidum  exemplifies how life cycle studies can consolidate disparate observations of a fungal organism, whether from environmental sequences, vegetative mycelia or field specimens, resulting in holistic species concepts critical to the assessment of the dimensions of fungal diversity.

Hypoxylon pulicicidum sp. nov. (Ascomycota, Xylariales), a Pantropical Insecticide-Producing Endophyte