Background: Lentinula edodes (Berk.) is the second most productive mushroom in the world. It contains compounds effective for antiviral, antitumor, antioxidant and immune regulation. Although genomes have previously been reported for this species, a high-quality chromosome-level reference for L. edodes is unavailable. This hinders detailed investigation of population genetics, breeding history of strains and genes related to environmental stress responses. Results: A high-quality chromosome-level genome was constructed. We separated a monokaryon from protoplasts of the commercial L. edodes strain L808 and assembled the genome of L. edodes using PacBio long-read and Illumina short-read sequencing, along with the high-throughput chromatin conformation capture (Hi-C) technique. We assembled a 45.87 Mb genome, and 99% of the sequences were anchored onto 10 chromosomes. The contig and scaffold N50 length were 2.17 and 4.94 Mb, respectively. Over 96% of the complete Benchmarking Universal Single-Copy Orthologs (BUSCO) were identified, and 9853 protein-coding genes were predicted. We performed population genome resequencing using 34 wild strains and 65 commercial cultivars of L. edodes originating from China, Japan, the United States and Australia. Based on whole-genome variants, we showed substantial differences in the Chinese wild population, which divided into different branches according to the main areas of their geographical distribution. We also determined the breeding history of L. edodes at the molecular level, and demonstrated that the cultivated strains in China mainly originated from wild strains from China and Northeast Asia. Phenotypic analysis showed that 99 strains exhibited differences on the Cd accumulation. Three significant loci in the of L. edodes genome were identified using the genome-wide association study (GWAS) of Cd accumulation traits. Functional genes associated with Cd accumulation traits were related to DNA ligase and aminoacyl tRNA synthetase, indicating that DNA damage repair and in vivo protein translation may be responses to Cd stress. Conclusions: A high-quality chromosome-level genome and population genetic data of L. edodes provide genetic resources for functional genomic, evolutionary and artificial breeding studies for L. edodes.

Traditional seed and fruit phenotyping are mainly accomplished by manual measurement or extraction of morphological properties from two-dimensional images. These methods are not only in low-throughput but also unable to collect their three-dimensional (3D) characteristics and internal morphology. X-ray computed tomography (CT) scanning, which provides a convenient means of non-destructively recording the external and internal 3D structures of seeds and fruits, offers a potential to overcome these limitations. However, the current CT equipment cannot be adopted to scan seeds and fruits with high throughput. And there is no specialized software for automatic extraction of phenotypes from CT images. Here, we introduced a high-throughput image acquisition approach by mounting a specially designed seed-fruit container onto the scanning bed. The corresponding 3D image analysis software, 3DPheno-Seed&Fruit, was created for automatic segmentation and rapid quantification of eight morphological phenotypes of internal and external compartments of seeds and fruits. 3DPheno-Seed&Fruit is a graphical user interface design and user-friendly software with an excellent phenotype result visualization function. We described the software in detail and benchmarked it based upon CT image analyses in seeds of soybean, wheat, peanut, pine nut, pistachio nut and dwarf Russian almond fruit. R² values between the extracted and manual measurements of seed length, width, thickness, and radius ranged from 0.80 to 0.96 for soybean and wheat. High correlations were found between the 2D (length, width, thickness, and radius) and 3D (volume and surface area) phenotypes for soybean. Overall, our methods provide robust and novel tools for phenotyping the morphological seed and fruit traits of various plant species, which could benefit crop breeding and functional genomics.

Morels (Morchella spp.) are popular edible fungi with significant economic and scientific value. However, white mold disease, caused by Paecilomyces penicillatus, can reduce morel yield by up to 80% in the main cultivation area in China. Paecilomyces is a polyphyletic genus and the exact phylogenetic placement of P. penicillatus is currently still unclear. Here, we obtained the first high-quality genome sequence of P. penicillatus generated through the single-molecule real-time (SMRT) sequencing platform. The assembled draft genome of P. penicillatus was 40.2 Mb, had an N50 value of 2.6 Mb and encoded 9454 genes. Phylogenetic analysis of single-copy orthologous genes revealed that P. penicillatus is in Hypocreales and closely related to Hypocreaceae, which includes several genera exhibiting a mycoparasitic lifestyle. CAZymes analysis demonstrated that P. penicillatus encodes a large number of fungal cell wall degradation enzymes. We identified many gene clusters involved in the production of secondary metabolites known to exhibit antifungal, antibacterial, or insecticidal activities. We further demonstrated through dual culture assays that P. penicillatus secretes certain soluble compounds that are inhibitory to the mycelial growth of Morchella sextelata. This study provides insights into the correct phylogenetic placement of P. penicillatus and the molecular mechanisms that underlie P. penicillatus pathogenesis.

Ganoderma tsugae is an endemic medicinal mushroom in Northeast China, providing important source of pharmaceutical product. Comparing with other Ganoderma species, wild G. tsugae can utilize coniferous wood. However, functional genes related to medicinal component synthesis and the genetic mechanism of conifer substrate utilization is still obscure. Here, we assembled a high-quality G. tsugae genome with 18 contigs and 98.5% BUSCO genes and performed the comparative genomics with other Ganoderma species. G. tsugae diverged from their common ancestor of G. lingzhi and G. sinense about 21 million years ago. Genes in G. tsugae-specific and G. tsugae-expanded gene families, such as salh, phea, cyp53a1, and cyp102a, and positively selected genes, such as glpk and amie, were functionally enriched in plant-pathogen interaction, benzoate degradation, and fanconi anemia pathway. Those functional genes might contribute to conifer substrate utilization of G. tsugae. Meanwhile, gene families in the terpene synthesis were identified and genome-wide SNP variants were detected in population. Finally, the study provided valuable genomic resources and offered useful hints for the functional gene mapping and investigation of key gene contributing to conifer cultivation substrate utilization and medicinal component biosynthesis.

Molecular barcoding and morphological characters were used to identify a new saprotrophic species in Pestalotiopsis, which was associated with senescent leaves of Eleutherococcus brachypus (Araliaceae) in Jilin Province, China. The matrix of the internal transcribed spacer (ITS) region, translation elongation factor 1-alpha (tef1-α), and β-tubulin (tub2) were used in the maximum likelihood (ML), maximum parsimony (MP) and Bayesian inference (BI) methods. The new collections formed a distinct clade with Pestalotiopsis lijiangensis. The new species differs from P. lijiangensis by its conidial length/width ratio. Detailed description and micrographs revealed that the species is unique in its olivaceous concolorous median cells and has significantly smaller conidia compared to other related species. The position of the apical appendages of Pestalotiopsis eleutherococci are distinct and are slightly shorter while the basal appendage is slightly longer compared to P. lijiangensis. Therefore, we introduce Pestalotiopsis eleutherococci as a novel species.

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.