Functional characterisation of genes putatively involved in fruiting body formation of the cultivated edible mushroom Agrocybe aegerita

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On the molecular level, fruiting body formation by edible mushrooms is poorly understood. As a basis of fruiting body formation in basidiomycetes, sexual development is well studied in the split gill mushroom Schizophyllum commune and the ink cap Coprinopsis cinerea. In contrast to that, edible mushrooms are up to now less accessible with regard to functional genetics approaches. Agrocybe aegerita is a commercially cultivated edible mushroom which exhibits the rare feature of monokaryotic fruiting sensu stricto which means that it is able to fruit without any sexual mating beforehand. A recently published work exploring molecular tools for transformation and constitutive gene expression in A. aegerita now permits functional genetics-based approaches in this mushroom. Within the proposed project, up to now uncharacterised fruiting-related genes (FRGs) will be functionally characterised, starting with four genes, the differential expression of which during fruiting has already been validated and which are predicted to likely encode transcription factors. In addition to the already set up functional-genetics tools for constitutive gene of interest expression (‘overexpression’) from preliminary work, CRISPR/Cas9-based gene deletion is to be adapted for A. aegerita. Ultimately, commonalities and differences between exceptional monokaryotic and regular dikaryotic fruiting of A. aegerita will be defined. To achieve this, RNA-sequencing is to be conducted, employing wild type monokaryotic fruiting stages versus different monokaryotic fruiting phenotypes exhibited by two selected mutants generated within this project. This approach will also help to understand the mutagenised genes’ roles within the complex genetic network of predicted FRGs in A. aegerita. Using this model system, the expected results will further bridge gaps in the knowledge on molecular biology of fruiting body formation and will also provide first insights on what differentiates monokaryotic fruiting from regular dikaryotic fruiting in mushrooms.