Anaerobic fungi exhibit a broad biotechnological potential. Nevertheless, there are significant knowledge gaps and many fundamental scientific questions on them, impeding methodological progress for the detection and especially maintenance of long-term cultures. We focus on basic issues in anaerobic fungi research in order to better understand their diversity, metabolic requirements and syntrophic microbial interactions. Thus, four distinct work packages were designed (https://www.hipoaf.com/).

1.      Standardization (University of Innsbruck)

This work package includes a centralized media preparation and culture collection maintenance by the University of Innsbruck and their distribution among the partners.

2.      Diversity and Cultivation (University of Innsbruck, Technical University of Munich)

To expand the knowledge on anaerobic fungal diversity, a database on their distribution shall be created and potential novel ecological niches for strain isolation shall be identified. Novel and known habitats shall then be screened for different representatives of Neocallimastigomycota as well as for their resting stages and potential inducers thereof. A special focus shall be laid on the identification and investigation of syntrophic interactions between anaerobic fungi and closely connected bacteria or archaea.

3.      Detection and Classification (Bavarian State Research Center)

Established (qPCR, visualization) and potentially novel (FISH, live/dead staining, PCR-based techniques, metagenomics/- transcriptomics and direct RNAseq ) detection and classification techniques for anaerobic fungi in environmental as well as experimental samples shall be validated and improved. Alongside, an optimized bioinformatics workflow for taxonomic and functional analyses shall be established, that can then be used to describe and phylogenetically assign novel strains.

4.      Semi-continuous Cultivation (Zurich University of Applied Sciences)

Biochemical growth requirements and cultivation parameters shall be defined with the aim to improve flow-through conditions. Therefore, suitable minimal as well as complex media for different habitats shall be defined and metabolites, growth inhibitors and promoters shall be investigated. Finally, a semi-continuous fermenter digesting lignocellulosic biomass by anaerobic fungi shall be constructed and operated.