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Abstract Members of the familyTrichomeriaceae,belonging to theChaetothyrialesorder and theAscomycotaphylum, are known for their capability to inhabit hostile environments characterized by extreme temperatures, oligotrophic conditions, drought, or presence of toxic compounds. The genusKnufiaencompasses many polyextremophilic species. In this report, the genomic and morphological features of the strain FJI-L2-BK-P2 presented, which was isolated from the Mars 2020 mission spacecraft assembly facility located at the Jet Propulsion Laboratory in Pasadena, California. The identification is based on sequence alignment for marker genes, multi-locus sequence analysis, and whole genome sequence phylogeny. The morphological features were studied using a diverse range of microscopic techniques (bright field, phase contrast, differential interference contrast and scanning electron microscopy). The phylogenetic marker genes of the strain FJI-L2-BK-P2 exhibited highest similarities with type strain ofKnufia obscura(CBS 148926^T) that was isolated from the gas tank of a car in Italy. To validate the species identity, whole genomes of both strains (FJI-L2-BK-P2 and CBS 148926^T) were sequenced, annotated, and strain FJI-L2-BK-P2 was confirmed asK. obscura.The morphological analysis and description of the genomic characteristics ofK. obscuraFJI-L2-BK-P2 may contribute to refining the taxonomy ofKnufiaspecies. Key morphological features are reported in thisK. obscurastrain, resembling microsclerotia and chlamydospore-like propagules. These features known to be characteristic features in black fungi which could potentially facilitate their adaptation to harsh environments.