Reintroducing Akanthomyces ampullifer: providing genetic barcodes, culture, and updated description for the dipteran pathogen rediscovered in Germany

The genus Akanthomyces (Ascomycota, Hypocreales) includes entomopathogenic species known to infect a variety of insects and spiders. In this study, we present the first isolate of A. ampullifer characterized by molecular methods, found on dead bodies of the common cave limoniid Limonia nubeculosa (Diptera) in the subterranean spaces of Southwestern Germany. In total, seven specimens exhibited distinctive morphological traits that, when compared with historical records, confirm their identification as A. ampullifer—particularly noted for its affinity to dipteran hosts. Absent from culture collections and molecular repositories, this species has eluded detailed scientific documentation using modern methods. Our research bridges this knowledge gap, providing the first genetic identification barcodes of five genes, living culture, cultivation requirements, and an updated description. This overlooked fungus is phylogenetically most closely related to the species A. pyralidarum, A. laosensis ..., , , # Reintroducing *Akanthomyces ampullifer*: providing genetic barcodes, culture, and updated description for the dipteran pathogen rediscovered in Germany

[https://doi.org/10.5061/dryad.cz8w9gjck](https://doi.org/10.5061/dryad.cz8w9gjck)

## Description of file structure

**ITS LSU TEF RPB1 RPB2 alignment final.fast**a

Concatenated alignment of ITS (1-563 bp), LSU (564-1404 bp), TEF (1405-2361 bp), RPB2 (2362-3082 bp), RPB2 (3083-4083 bp)**Akanthomyces Tree edited.pdf**

The best-scoring maximum likelihood tree was calculated from ITS, LSU, TEF1-α, RPB1, and RPB2 loci sequences showing species relationships within the genus Akanthomyces and the position of A. ampullifer CCF 6648. Maximum likelihood bootstrap supports are attached to nodes, with only values >70 % shown; lower supports are indicated with a hyphen; ex-type strains and type specimens are designated by a superscript T; the tree is rooted with Samsoniella species

**Tree_Akanthomyces_5G.newick**

The best-scoring maximum ...