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The Cys2His2 zinc finger protein gene c2h2 of Schizophyllum commune is involved in mushroom formation. Its inactivation results in a strain that is arrested at the stage of aggregate formation. In this study, the c2h2 orthologue of Agaricus bisporus was over-expressed in this white button mushroom forming basidiomycete using Agrobacterium-mediated transformation. Morphology, cap expansion rate, and total number and biomass of mushrooms were not affected by over-expression of c2h2. However, yield per day of the c2h2 over-expression strains peaked 1 day earlier. These data and expression analysis indicate that C2H2 impacts timing of mushroom formation at an early stage of development, making its encoding gene a target for breeding of commercial mushroom strains. Electronic supplementary material The online version of this article (doi:10.1007/s00253-016-7574-9) contains supplementary material, which is available to authorized users.

Recent genome-wide studies have demonstrated that fungi possess the machinery to alternatively splice pre-mRNA. However, there has not been a systematic categorization of the functional impact of alternative splicing in a fungus. We investigate alternative splicing and its functional consequences in the model mushroom forming fungus Schizophyllum commune. Alternative splicing was demonstrated for 2,285 out of 12,988 expressed genes, resulting in 20% additional transcripts. Intron retentions were the most common alternative splicing events, accounting for 33% of all splicing events, and 43% of the events in coding regions. On the other hand, exon skipping events were rare in coding regions (1%) but enriched in UTRs where they accounted for 57% of the events. Specific functional groups, including transcription factors, contained alternatively spliced genes. Alternatively spliced transcripts were regulated differently throughout development in 19% of the 2,285 alternatively spliced genes. Notably, 69% of alternatively spliced genes have predicted alternative functionality by loss or gain of functional domains, or by acquiring alternative subcellular locations. S. commune exhibits more alternative splicing than any other studied fungus. Taken together, alternative splicing increases the complexity of the S. commune proteome considerably and provides it with a rich repertoire of alternative functionality that is exploited dynamically.

Establishment of the dikaryotic mycelium and formation of fruiting bodies are highly complex developmental programmes that are activated by a combination of environmental cues. A wide variety of proteins are expected to regulate and coordinate these programmes or to fulfil enzymatic conversions or structural roles. With the identification of the first genes involved in mushroom development, we are only at the beginning of understanding fruiting body formation. The process of identification of genes will be accelerated by whole genome expression studies and increased availability of molecular tools to assign functions to genes. Establishment of the dikaryon and emergence of fruiting bodies in basidiomycetes are regulated by the mating-type genes. These genes encode DNA-binding proteins and pheromones and their receptors. Regulation of fruiting by the mating-type genes is mediated by downstream transcription factors. Several genes encoding such regulatory proteins have now been identified. Regulatory circuits ultimately activate genes encoding structural proteins or enzymes that are involved in fruiting body formation. The role of hydrophobins is well established. They enable hyphae to escape the aqueous environment to allow fruiting body development. Moreover, they coat aerial structures and line air channels in mushrooms. The hydrophobic coating irreversibly directs growth of hyphae into the air, allows dispersal of spores and ensures gas exchange in fruiting bodies under humid conditions. Apart from hydrophobins, phenolics polymerised by the action of laccases may contribute to surface hydrophobicity of fruiting bodies. These enzymes have also been proposed to cross-link cell walls of hyphae in the fruiting bodies but this still has to be established. Experimental evidence indicates that cytochrome P450 enzymes, lectins, haemolysins and expansins also function in mushroom development. Lectins may be involved in aggregation of hyphae, haemolysins in signalling particularly to induce apoptosis of selected hyphae in the fruiting body, while expansins may be involved in cell wall modification and extension.

Lecanicillium fungicola causes dry bubble disease and is an important problem in the cultivation of Agaricus bisporus. Little is known about the defense of mushrooms against pathogens in general and L. fungicola in particular. In plants and animals, a first attack by a pathogen often induces a systemic response that results in an acquired resistance to subsequent attacks by the same pathogen. The development of functionally similar responses in these two eukaryotic kingdoms indicates that they are important to all multi-cellular organisms. We investigated if such responses also occur in the interaction between the white button mushroom and L. fungicola. A first infection of mushrooms of the commercial A. bisporus strain Sylvan A15 by L. fungicola did not induce systemic resistance against a subsequent infection. Similar results were obtained with the A. bisporus strain MES01497, which was demonstrated to be more resistant to dry bubble disease. Apparently, fruiting bodies of A. bisporus do not express induced resistance against L. fungicola.

Lecanicillium fungicola causes dry bubble disease in commercially cultivated mushroom. This review summarizes current knowledge on the biology of the pathogen and the interaction between the pathogen and its most important host, the white-button mushroom, Agaricus bisporus. The ecology of the pathogen is discussed with emphasis on host range, dispersal and primary source of infection. In addition, current knowledge on mushroom defence mechanisms is reviewed. Taxonomy:Lecanicillium fungicola (Preuss) Zare and Gams: Kingdom Fungi; Phylum Ascomycota; Subphylum Pezizomycotina; Class Sordariomycetes; Subclass Hypocreales; Order Hypocreomycetidae; Family Cordycipitaceae; genus Lecanicillium. Host range:Agaricus bisporus, Agaricus bitorquis and Pleurotus ostreatus. Although its pathogenicity for other species has not been established, it has been isolated from numerous other basidiomycetes. Disease symptoms: Disease symptoms vary from small necrotic lesions on the caps of the fruiting bodies to partially deformed fruiting bodies, called stipe blow-out, or totally deformed and undifferentiated masses of mushroom tissue, called dry bubble. The disease symptoms and severity depend on the time point of infection. Small necrotic lesions result from late infections on the fruiting bodies, whereas stipe blow-out and dry bubble are the result of interactions between the pathogen and the host in the casing layer. Economic importance:Lecanicillium fungicola is a devastating pathogen in the mushroom industry and causes significant losses in the commercial production of its main host, Agaricus bisporus. Annual costs for mushroom growers are estimated at 2–4% of total revenue. Reports on the disease originate mainly from North America and Europe. Although China is the main producer of white-button mushrooms in the world, little is known in the international literature about the impact of dry bubble disease in this region. Control: The control of L. fungicola relies on strict hygiene and the use of fungicides. Few chemicals can be used for the control of dry bubble because the host is also sensitive to fungicides. Notably, the development of resistance of L. fungicola has been reported against the fungicides that are used to control dry bubble disease. In addition, some of these fungicides may be banned in the near future.

BACKGROUND: Mycelia of higher fungi consist of interconnected hyphae that are compartmentalized by septa. These septa contain large pores that allow streaming of cytoplasm and even organelles. The cytoplasm of such mycelia is therefore considered to be continuous. METHODOLOGY/PRINCIPAL FINDINGS: Here, we show by laser dissection that septa of Schizophyllum commune can be closed depending on the environmental conditions. The most apical septum of growing hyphae was open when this basidiomycete was grown in minimal medium with glucose as a carbon source. In contrast, the second and the third septum were closed in more than 50% and 90% of the cases, respectively. Interestingly, only 24 and 37% of these septa were closed when hyphae were growing in the absence of glucose. Whether a septum was open or closed also depended on physical conditions of the environment or the presence of toxic agents. The first septum closed when hyphae were exposed to high temperature, to hypertonic conditions, or to the antibiotic nourseothricin. In the case of high temperature, septa opened again when the mycelium was placed back to the normal growth temperature. CONCLUSIONS/SIGNIFICANCE: Taken together, it is concluded that the septal pores of S. commune are dynamic structures that open or close depending on the environmental conditions. Our findings imply that the cytoplasm in the mycelium of a higher fungus is not continuous per se.

Disruption of the SC3 gene in the basidiomycete Schizophyllum commune affected not only formation of aerial hyphae but also attachment to hydrophobic surfaces. However, these processes were not completely abolished, indicating involvement of other molecules. We here show that the SC15 protein mediates formation of aerial hyphae and attachment in the absence of SC3. SC15 is a secreted protein of 191 aa with a hydrophilic N-terminal half and a highly hydrophobic C-terminal half. It is not a hydrophobin as it lacks the eight conserved cysteine residues found in these proteins. Besides being secreted into the medium, SC15 was localized in the cell wall and the mucilage that binds aerial hyphae together. In a strain in which the SC15 gene was deleted (DeltaSC15) formation of aerial hyphae and attachment were not affected. However, these processes were almost completely abolished when the SC15 gene was deleted in the DeltaSC3 background. The absence of aerial hyphae in the DeltaSC3DeltaSC15 strain can be explained by the inability of the strain to lower the water surface tension and to make aerial hyphae hydrophobic.