Cladobotryum protrusum is one of the mycoparasites that cause cobweb disease on cultivated edible mushrooms. However, the molecular mechanisms of evolution and pathogenesis of C. protrusum on mushrooms are largely unknown. Here, we report a high-quality genome sequence of C. protrusum using the single-molecule, real-time sequencing platform of PacBio and perform a comparative analysis with closely related fungi in the family Hypocreaceae. The C. protrusum genome, the first complete genome to be sequenced in the genus Cladobotryum, is 39.09 Mb long, with an N50 of 4.97 Mb, encoding 11,003 proteins. The phylogenomic analysis confirmed its inclusion in Hypocreaceae, with its evolutionary divergence time estimated to be ~170.1 million years ago. The genome encodes a large and diverse set of genes involved in secreted peptidases, carbohydrate-active enzymes, cytochrome P450 enzymes, pathogen–host interactions, mycotoxins, and pigments. Moreover, C. protrusum harbors arrays of genes with the potential to produce bioactive secondary metabolites and stress response-related proteins that are significant for adaptation to hostile environments. Knowledge of the genome will foster a better understanding of the biology of C. protrusum and mycoparasitism in general, as well as help with the development of effective disease control strategies to minimize economic losses from cobweb disease in cultivated edible mushrooms.
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.
Grifola frondosa is an economically important edible and medicinal mushroom usually produced on substrate consisting of sawdust supplemented with wheat bran. Cultivation of G. frondosa on crop straw (corn cob, corn straw, rice straw, and soybean straw) as a substrate was optimized by using the D-optimum method of the simplex-lattice design, and the alternative of crop straw as a substitute for sawdust in the substrate composition was determined by the optimized model. The results showed that there was a significant positive correlation existing between the yield and corn cob. The growth cycle was negatively correlated with sawdust, corn cob and soybean straw, with sawdust significantly shortening the growth cycle of G. frondosa. The optimized high-yielding formula included 73.125% corn cob, 1.875% rice straw, 23% wheat bran and 2% light calcium carbonate (CaCO3) (C/N = 48.40). The average yield of the first flush was 134.72 ± 4.24 g/bag, which was increased by 39.97% compared with the control formula. The biological efficiency (BE) was 44.91 ± 1.41%, which was increased by 38.53% compared with the control. Based on the results of this study, corn cob can replace sawdust as one of the main cultivation substrates of G. frondosa.
Mycogone perniciosa causes wet bubble disease in Agaricus bisporus and various Agaricomycetes species. In a previous work, we identified 41 GH18 chitinase genes and other pathogenicity-related genes in the genome of M. perniciosa Hp10. Chitinases are enzymes that degrade chitin, and they have diverse functions in nutrition, morphogenesis, and pathogenesis. However, these important genes in M. perniciosa have not been fully characterized, and their functions remain unclear. Here, we performed a genome-wide analysis of M. perniciosa GH18 genes and analyzed the transcriptome profiles and GH18 expression patterns in M. perniciosa during the time course of infection in A. bisporus. Phylogenetic analysis of the 41 GH18 genes with those of 15 other species showed that the genes were clustered into three groups and eight subgroups based on their conserved domains. The GH18 genes clustered in the same group shared different gene structures but had the same protein motifs. All GH18 genes were localized in different organelles, were unevenly distributed on 11 contigs, and had orthologs in the other 13 species. Twelve duplication events were identified, and these had undergone both positive and purifying selection. The transcriptome analyses revealed that numerous genes, including transporters, cell wall degrading enzymes (CWDEs), cytochrome P450, pathogenicity-related genes, secondary metabolites, and transcription factors, were significantly upregulated at different stages of M. perniciosa Hp10 infection of A. bisporus. Twenty-three out of the 41 GH18 genes were differentially expressed. The expression patterns of the 23 GH18 genes were different and were significantly expressed from 3 days post-inoculation of M. perniciosa Hp10 in A. bisporus. Five differentially expressed GH18 genes were selected for RT-PCR and gene cloning to verify RNA-seq data accuracy. The results showed that those genes were successively expressed in different infection stages, consistent with the previous sequencing results. Our study provides a comprehensive analysis of pathogenicity-related and GH18 chitinase genes’ influence on M. perniciosa mycoparasitism of A. bisporus. Our findings may serve as a basis for further studies of M. perniciosa mycoparasitism, and the results have potential value for improving resistance in A. bisporus and developing efficient disease-management strategies to mitigate wet bubble disease.
cornea cv. Yu Muer) is a new white variety of edible fungus that was selected from a mutant of Auricularia cornea by the Engineering Research Center of the Ministry of Education, Jilin Agricultural University. Yu Muer is in genus Auricularia, family Auriculariaceae, order Auriculariales, class Agaricomycetes, and phylum Basidiomycota. It is an edible fungus and is also used in medicine (Royse, 2014; Wang, Jiang, et al., 2019a; Wang, Li, et al., 2019b). The fruiting bodies of Yu Muer are thick, tender, and crispy tastes like jellyfish, and have a jade-like warm, soft color. It is rich in nutrients, including physiologically active substances such as polysaccharides,
Brown blotch disease (BBD) caused by Pseudomonas tolaasii is one of the most devastating diseases of Pleurotus spp. worldwide. Breeding for resistant strains is the most effective method for controlling BBD. To identify resistant germplasm for BBD management, 97 strains comprising 21 P. cf. floridanus, 20 P. ostreatus, and 56 P. pulmonarius were screened by two different methods; namely, inoculation of the pathogen on the mushroom pileus (IMP) and on the spawned substrate (IMSS) under controlled conditions. Out of the 97 strains screened, 22 P. pulmonarius, and four P. cf. floridanus were moderately resistant to BBD using the IMP method. Eleven P. pulmonarius, six P. cf. florida, and one P. ostreatus strains were highly resistant to BBD using the IMSS method. All of the 97 strains showed varying degrees of susceptibility using the IMP method, but eight strains were completely resistant using the IMSS method. Combining these two methods, five strains were highly resistant (four P. pulmonarius and one P. cf. floridanus) and 11 were moderately resistant (eight P. pulmonarius and three P. cf. floridanus). The resistance sources to P. tolaasii identified in P. pulmonarius and P. cf. floridanus could be used for further breeding of Pleurotus spp.
Ewingella americana is a cosmopolitan bacterial pathogen that has been isolated from many hosts. Here, we sequenced a high-quality genome of E. americana B6-1 isolated from Flammulina filiformis, an important cultivated mushroom, performed a comparative genomic analysis with four other E. americana strains from various origins, and tested the susceptibility of B6-1 to antibiotics. The genome size, predicted genes, and GC (guanine-cytosine) content of B6-1 was 4.67 Mb, 4301, and 53.80%, respectively. The origin of the strains did not significantly affect the phylogeny, but mobile genetic elements shaped the evolution of the genus Ewingella. The strains encoded a set of common genes for type secretion, virulence effectors, CAZymes, and toxins required for pathogenicity in all hosts. They also had antibiotic resistance, pigments to suppress or evade host defense responses, as well as genes for adaptation to different environmental conditions, including temperature, oxidation, and nutrients. These findings provide a better understanding of the virulence, antibiotic resistance, and host adaptation strategies of Ewingella, and they also contribute to the development of effective control strategies.
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