Cucurbitariaceae has a high biodiversity worldwide on various hosts and is distributed in tropical and temperate regions. Woody litters collected in Changchun, Jilin Province, China, revealed a distinct collection of fungi in the family Cucurbitariaceae based on morphological and molecular data. Phylogenetic analyses of the concatenated matrix of the internal transcribed spacer (ITS) region, the large subunit (LSU) of ribosomal DNA, the RNA polymerase II subunit (rpb2), the translation elongation factor 1-alpha (tef1-α) and β-tubulin (β-tub) genes indicated that the isolates represent Allocucurbitaria and Parafenestella species based on maximum likelihood (ML), maximum parsimony (MP) and Bayesian analysis (BPP). We report four novel species: Allocucurbitaria mori, Parafenestella changchunensis, P. ulmi and P. ulmicola. The importance of five DNA markers for species-level identification in Cucurbitariaceae was determined by Assemble Species by Automatic Partitioning (ASAP) analyses. The protein-coding gene β-tub is determined to be the best marker for species level identification in Cucurbitariaceae.
Identification of monokaryons and their mating types and discrimination of hybrid offspring are key steps for the crossbreeding of Pleurotus tuoliensis (Bailinggu). However, conventional crossbreeding methods are troublesome and time consuming. Using RNA-seq technology, we developed new expressed sequence tag-simple sequence repeat (EST-SSR) markers for Bailinggu to easily and rapidly identify monokaryons and their mating types, genetic diversity and hybrid offspring. We identified 1110 potential EST-based SSR loci from a newly-sequenced Bailinggu transcriptome and then randomly selected 100 EST-SSRs for further validation. Results showed that 39, 43 and 34 novel EST-SSR markers successfully identified monokaryons from their parent dikaryons, differentiated two different mating types and discriminated F1 and F2 hybrid offspring, respectively. Furthermore, a total of 86 alleles were detected in 37 monokaryons using 18 highly informative EST-SSRs. The observed number of alleles per locus ranged from three to seven. Cluster analysis revealed that these monokaryons have a relatively high level of genetic diversity. Transfer rates of the EST-SSRs in the monokaryons of closely-related species Pleurotuseryngii var. ferulae and Pleurotus ostreatus were 72% and 64%, respectively. Therefore, our study provides new SSR markers and an efficient method to enhance the crossbreeding of Bailinggu and closely-related species.
Cladobotryum dendroides, which causes cobweb disease in edible mushrooms, is one of the major fungal pathogens. Our previous studies focused on the genetic and morphological characterization of this fungus, as well as its pathogenicity and the identification of appropriate fungicides. However, little is known about the genome characters, pathogenic genes, and molecular pathogenic mechanisms of C. dendroides. Herein, we reported a high-quality de novo genomic sequence of C. dendroides and compared it with closely-related fungi. The assembled C. dendroides genome was 36.69 Mb, consisting of eight contigs, with an N50 of 4.76 Mb. This genome was similar in size to that of C. protrusum, and shared highly conserved syntenic blocks and a few inversions with C. protrusum. Phylogenetic analysis revealed that, within the Hypocreaceae, Cladobotryum was closer to Mycogone than to Trichoderma, which is consistent with phenotypic evidence. A significant number of the predicted expanded gene families were strongly associated with pathogenicity, virulence, and adaptation. Our findings will be instrumental for the understanding of fungi–fungi interactions, and for exploring efficient management strategies to control cobweb disease.
Wet bubble disease, caused by Mycogone perniciosa, is a major threat to Agaricus bisporus production in China. In order to understand the variability 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, molecular, and 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 range 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.
In order to reveal the genetic variation signals of Auricularia heimuer that have occurred during their domestication and to find potential functional gene families, we constructed a monokaryotic pan-genome of A. heimuer representing four cultivated strains and four wild strains. The pan-genome contained 14,089 gene families, of which 67.56% were core gene families and 31.88% were dispensable gene families. We screened substrate utilization-related genes such as the chitinase gene ahchi1 of the glycoside hydrolase (GH) 18 family and a carbohydrate-binding module (CBM)-related gene from the dispensable families of cultivated populations. The genomic difference in the ahchi1 gene between the wild and cultivated genomes was caused by a 33 kb presence/absence variation (PAV). The detection rate of the ahchi1 gene was 93.75% in the cultivated population, significantly higher than that in the wild population (17.39%), indicating that it has been selected in cultivated strains. Principal component analysis (PCA) of the polymorphic markers in fragments near the ahchi1 gene was enriched in cultivated strains, and this was caused by multiple independent instances of artificial selection. We revealed for the first time the genetic basis of the ahchi1 gene in domestication, thereby providing a foundation for elucidating the potential function of the ahchi1 gene in the breeding of A. heimuer.
Introduction: Tomato growers need strategies that improve fruit nutritional quality and soil health while reducing dependence on synthetic fertilizers and wasting less crop straw. Fungal-fermented straw products (FSP) are a candidate amendment, but their suitable application rate, placement depth and side effects on soil microbes and antibiotic resistance genes (ARGs) remain unclear. Methods: An FSP was produced from corn straw via solid-state fermentation for 30 d using Auricularia cornea cv. Yumuer. A single-season greenhouse pot experiment was conducted with an unamended control (no FSP) and six FSP treatments combining different rates (0.5–5% w/w) and soil incorporation depths (3 and 10 cm). Measurements included tomato yield (growth and fruit yield), fruit quality, soil health, biochemical responses, and molecular responses. Results: FSP at 2% (w/w) incorporated to 10 cm increased yield by about 30% and raised fruit lycopene and vitamin C by 20–40% compared with the control. It also enhanced soil organic carbon, available P and K, humic substances and key enzymes, and shifted microbes toward decomposers and plant-beneficial taxa (e.g., Streptomyces , Sphingomonas , Nocardioides , and Arthrobacter ). A high surface dose (5% at 3 cm) increased total ARGs, whereas 2% at 10 cm achieved quality and soil benefits with ARG levels comparable to or lower than the control. Discussion: These results suggest a practical application pattern that balances agronomic benefits with ARG-related risk for recycling crop residues in intensive tomato systems.