Wet bubble disease (WBD) in Agaricus bisporus caused by Mycogone species imposes a substantial economic loss to mushroom production in China. Currently, fungicide application is the main method to control WBD. However, excessive use of fungicide is challenged by the appearance of resistance and food safety. Therefore, it is necessary to explore safe and efficient strategies to control WBD. Strain 9-13, isolated from the rhizosphere soil of Taxus chinensis, showed strong inhibitory activity against three Mycogone species. According to morphological and biochemical characteristics, and multilocus phylogenetic analysis, the strain was identified as Streptomyces nojiriensis. In addition, strain 9-13 extracts significantly inhibited mycelial growth and spore germination of M. perniciosa, M. rosea and M. xinjiangensis in vitro. Strain 9-13 and its extracts also exhibited broad-spectrum antifungal activities against 12 selected plant pathogenic fungi. Scanning electron microscopic observations showed that extracts destroyed mycelial structure, inducing mycelia to twist and shrink. Moreover, transmission electron microscopy revealed that extracts resulted in severe plasmolysis, rupture of cell membrane and a decrease in cell inclusions, and the cell wall appeared a rough and uneven surface. Notably, the extracts obviously reduced disease severity and incidence of WBD by from 83.85% to 87.32% in fruiting bodies and 77.36% in mushroom beds, and maintained fruiting time and color on harvested mushroom. Collectively, these results clearly indicate that S. nojiriensis 9-13 is a promising biocontrol agent to control WBD on A. bisporus.

Agaricus bisporus (Lange) Imbach is the most widely cultivated mushroom in the world. A. bisporus wet bubble disease is one of the most severe diseases of white button mushrooms and is caused by the fungal pathogen Hypomyces perniciosus. The pathogen causes a drastic reduction in mushroom yield because of malformation and deterioration of the basidiomes. However, the mechanism of the button mushroom\'s malformation development after infection with H. perniciosus remains obscure. Therefore, to reveal the mechanism of A. bisporus malformation caused by H. perniciosus, the interaction between the pathogen and host was investigated in this study using histopathological, physiological, and transcriptomic analyses. Results showed that irrespective of the growth stages of A. bisporus basidiomes infected with H. perniciosus, the host\'s malformed basidiomes and enlarged mycelia and basidia indicated that the earlier the infection with H. perniciosus, the more the malformation of the basidiomes. Analyzing physiological and transcriptomic results in tandem, we concluded that H. perniciosus causes malformation development of A. bisporus mainly by affecting the metabolism level of phytohormones (N6-isopentenyladenosine, cis-zeatin, and N6-[delta 2-isopentenyl]-adenine) of the host\'s fruiting bodies rather than using toxins. Our findings revealed the mechanism of the button mushroom\'s malformation development after infection with H. perniciosus, providing a reference for developing realistic approaches to control mushroom diseases. Our results further clarified the interaction between A. bisporus and H. perniciosus and identified the candidate genes for A. bisporus wet bubble disease resistance breeding. Additionally, our work provides a valuable theoretical basis and technical support for studying the interaction between other pathogenic fungi and their fungal hosts.

White button mushroom, Agaricus bisporus (Lange) Imbach, is the most extensively cultivated and edible mushroom worldwide. The production of A. bisporus is commonly affected by wet bubble disease (WBD), imposing a significant economic burden in China. Although studies have shown that this disease is caused by fungi of the Mycogone genus, the pathogen has not been fully characterized. In this study, 802 samples of diseased fruiting bodies of A. bisporus were collected from nine major mushroom-cultivating provinces in China, yielding a total of 586 Mycogone isolates. The morphologic characteristics of these isolates were observed and compared, and multilocus phylogenetic analyses (internal transcribed spacer [ITS], ACT, TEF1-α, TUB, RPB2, and large ribosomal subunit [LSU]) were performed on the selected representative isolates. Three Mycogone species were identified: a new species, M. xinjiangensis; M. perniciosa; and M. rosea. Mycogone rosea was the first ever reported in China. Furthermore, M. rosea was found to be the most prevalent species (54.95% of all isolates) in all the sampled areas, except in Hubei and Xinjiang, followed by M. perniciosa (39.93%) and M. xinjiangensis (5.12%). Pathogenicity tests on the fruiting body and mushroom bed substantiated Koch\'s postulates by the development of mildly different symptoms after inoculation with each species. This study, therefore, enhances our knowledge of the species associated with WBD in A. bisporus and provides useful insights for preventing WBD and allied diseases.

Mycogone perniciosa is a mycoparasite causing Wet Bubble Diseases (WBD) of Agaricus bisporus. In the present study, the whole genome of M. perniciosa strain MgR1 was sequenced using Illumina NextSeq500 platform. This sequencing generated 8.03 Gb of high-quality data and a draft genome of 39 Mb was obtained through a de novo assembly of the high-quality reads. The draft genome resulted into prediction of 9276 genes from the 1597 scaffolds. NCBI-based homology analysis revealed the identification of 8660 genes. Notably, non-redundant protein database analysis of the M. perniciosa strain MgR1 revealed its close relation with the Trichoderma arundinaceum. Moreover, ITS-based phylogenetic analysis showed the highest similarity of M. perniciosa strain MgR1 with Hypomyces perniciosus strain CBS 322.22 and Mycogone perniciosa strain PPRI 5784. Annotation of the 3917 genes of M. perniciosa strain MgR1 grouped in three major categories viz. biological process (2583 genes), cellular component (2013 genes), and molecular function (2919 genes). UniGene analysis identified 2967 unique genes in M. perniciosa strain MgR1. In addition, prediction of the secretory and pathogenicity-related genes based on the fungal database indicates that 1512 genes (16% of predicted genes) encode for secretory proteins. Moreover, out of 9276 genes, 1296 genes were identified as pathogenesis-related proteins matching with 51 fungal and bacterial genera. Overall, the key pathogenic genes such as lysine M protein domain genes, G protein, hydrophobins, and cytochrome P450 were also observed. The draft genome of MgR1 provides an understanding of pathogenesis of WBD in A. bisporus and could be utilized to develop novel management strategies.

Wet bubble disease, caused by Mycogone perniciosa, is a major threat to Agaricus bisporus production in China. In order to understand the variability of in genetic, pathogenicity, morphology, and symptom production of the fungus, 18 isolates of the pathogen were collected from diseased A. bisporus in different provinces in China. The isolates were characterized by a combination of morphological, cultural, and molecular pathogenicity testing on different strains of A. bisporus and amplified fragment length polymorphism (AFLP) analysis. The 18 isolates were identified by Koch\'s postulate and confirmed different pathogenic variability among them. The yellow to brown isolates were more virulent than the white isolates. AFLP markers clustered the isolates into two distinct groups based on their colony color, with a high level of polymorphism of Jaccard similarities ranges from 0.39% to 0.64%. However, there was no evidence of an association between the genetic diversity and the geographical origin of the isolates. Through knowledge of the genetic diversity, phenotypic virulence of M. perniciosa is a key factor for successful breeding of resistant strains of A. bisporus and developing of an integrated disease management strategy to manage wet bubble disease of A. bisporus.

The mycoparasitic fungus Hypomyces perniciosus causes wet bubble disease of mushrooms, particularly Agaricus bisporus. The genome of a highly virulent strain of H. perniciosus HP10 was sequenced and compared to three other fungi from the order Hypocreales that cause disease on A. bisporus. H. perniciosus genome is ~44 Mb, encodes 10,077 genes and enriched with transposable elements up to 25.3%. Phylogenetic analysis revealed that H. perniciosus is closely related to Cladobotryum protrusum and diverged from their common ancestor ~156.7 million years ago. H. perniciosus has few secreted proteins compared to C. protrusum and Trichoderma virens, but significantly expanded protein families of transporters, protein kinases, CAZymes (GH 18), peptidases, cytochrome P450, and SMs that are essential for mycoparasitism and adaptation to harsh environments. This study provides insights into H. perniciosus evolution and pathogenesis and will contribute to the development of effective disease management strategies to control wet bubble disease.