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.

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.

Dietary alfalfa fiber (AF) is conceived to modulate gut microbial richness and diversity to improve the health and growth of weaning piglets. The objective of this study was to evaluate the prebiotic effects of AF on diarrhea incidence, the production of short-chain fatty acids (SCFAs), and microbiota composition in weaning piglets. This study utilized 100 crossbred piglets (Duroc × Landrace × Yorkshire) with a body weight of 8.42 ± 1.88 kg randomly assigned to the following treatments: 0.00% AF meal (A), 6.00% of AF meal (B), 12.00% AF meal (C), and 18.00% AF meal (D). The cecum samples were used to determine microbial community composition and diversity through high-throughput 16S rDNA sequencing. The results of this study show that the lowest average daily gain (ADG) was observed in treatment D, and the highest ADG was recorded in treatment C. However there was no significant difference between the treatment groups and the control. The average daily feed intake (ADFI) was significantly higher in treatment C compared to the other treatments. The feed conversion ratio was high in the control group compared to the AF treated groups. The highest diarrhea incidence was observed in treatment A and the lowest diarrhea incidence was observed in treatment C and D. The highest acetate and propionate levels were observed in treatment B, but there was no significant difference between the treatment groups and the control. The supplementation of AF significantly increased the butyrate level in treatment D compared with treatments A and B but was not significantly different from treatment C. The Observed_species richness and Simpson diversity values of the cecum bacterial composition in the AF fed piglets were higher than the control. In addition, the Chao 1 richness and Shannon diversity increased with an increase in AF supplementation, reaching a plateau at treatment B and C, then decreasing at treatment D. The Bacteroidetes, Firmicutes, Tenericutes, Proteobacteria, Cyanobacteria, Spirochaetae, Actinobacteria, Fibrobacteres, Saccharibacteria, Synergistetes, Chlamydiae, Elusimicrobia, Deferribacteres, Fusobacteria, and others were relatively abundant in all treatments. The Bacteroidetes and Firmicutes were the dominant phyla, accounting for 98% of all reads. AF treatment decreased the Bacteroidetes phylum and increased the Firmicutes phylum compared with treatment A. Therefore, the dietary inclusion of AF may decrease diarrhea incidence, increase cecal bacterial composition and richness, and consequently improve the growth performance of weaning piglets.

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.

Cobweb disease is a fungal disease that can cause serious damage to edible mushrooms worldwide. To investigate cobweb disease in Morchella sextelata in Guizhou Province, China, we isolated and purified the pathogen responsible for the disease. Through morphological and molecular identification and pathogenicity testing on infected M. sextelata, we identified Cladobotryum mycophilum as the cause of cobweb disease in this region. This is the first known occurrence of this pathogen causing cobweb disease in M. sextelata anywhere in the world. We then obtained the genome of C. mycophilum BJWN07 using the HiFi sequencing platform, resulting in a high-quality genome assembly with a size of 38.56 Mb, 10 contigs, and a GC content of 47.84%. We annotated 8428 protein-coding genes in the genome, including many secreted proteins, host interaction-related genes, and carbohydrate-active enzymes (CAZymes) related to the pathogenesis of the disease. Our findings shed new light on the pathogenesis of C. mycophilum and provide a theoretical basis for developing potential prevention and control strategies for cobweb disease.

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.