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The complete mitochondrial genome of the Nematophagous fungus Esteya vermicola CBS 115803 was determined using the PacBio RS II sequencing technology. The circular molecule is 47,282bp in length with a GC content of 24.85%. Annotated genes including 14 conserved protein-coding genes, the large and the small rRNA subunit (rnl and rns) and 27 tRNAs. The phylogenetic analysis showed that E. vermicola had close genetic relationship with the genus Sporothrix.

Nematophagous (NP) fungi are ecologically important components of the soil microbiome in natural ecosystems. Esteya vermicola (Ev) has been reported as a NP fungus with a poorly understood evolutionary history and mechanism of adaptation to parasitism. Furthermore, NP fungal genomic basis of lifestyle was still unclear. We sequenced and annotated the Ev genome (34.2 Mbp) and integrated genetic makeup and evolution of pathogenic genes to investigate NP fungi. The results revealed that NP fungi had some abundant pathogenic genes corresponding to their niche. A number of gene families involved in pathogenicity were expanded, and some pathogenic orthologous genes underwent positive selection. NP fungi with diverse morphological features exhibit similarities of evolutionary convergence in attacking nematodes, but their genetic makeup and microscopic mechanism are different. Endoparasitic NP fungi showed similarity in large number of transporters and secondary metabolite coding genes. Noteworthy, expanded families of transporters and endo-beta-glucanase implied great genetic potential of Ev in quickly perturbing nematode metabolism and parasitic behavior. These results facilitate our understanding of NP fungal genomic features for adaptation to nematodes and lay a solid theoretical foundation for further research and application.

The emerald ash borer (EAB, Agrilus planipennis), an ash-tree wood-boring beetle, has caused widespread mortality of ash. Asian ash, which coevolved with EAB, is considered more resistant than its North American and European congeners. Although some compounds and proteins related to resistance to EAB have been identified, the underlying ash resistance mechanism to EAB still needs further study. The Asian ash species, Fraxinus chinensis var. rhynchophylla, is highly resistant to EAB. In this study, metabolic and transcriptional profiling of the phloem of this species was investigated, and differentially expressed metabolites and genes were analyzed by comparing them with those of the susceptible F. pennsylvanica. Four hundred and twenty-eight metabolites were detected in both species, and several coumarins and lignans, which were exclusive to F. chinensis var. rhynchophylla, were identified. Compared with susceptible F. pennsylvanica, genes related to phenylpropanoid biosynthesis, ethylene (ET), and jasmonic acid (JA) biosynthesis and signaling in F. chinensis var. rhynchophylla were found to be up-regulated. It was hypothesized that coumarins, lignans, and ET and JA signaling might contribute to greater resistance to EAB in F. chinensis var. rhynchophylla. This study suggests candidate metabolites and genes for biomarker development in future ash-breeding programs.

Symbioses have played pivotal roles in biological, ecological, and evolutionary diversification. Symbiotic bacteria affect the biology of hosts in a number of ways. Esteya vermicola, an endoparasitic nematophagous fungus, has high infectivity in the pine wood nematode (PWN), which causes devastating ecological damage and economic losses in Asia and Europe. An integration of molecular, phylogenetic, and morphological analyses revealed that surface-sterilized E. vermicola with septate hyphae from different geographic locations harbor bacterial endosymbionts. 16S rRNA gene sequences from four fungal strains all clustered in a well-supported monophyletic clade that was the most closely related to Pseudomonas stutzeri and affiliated with Gammaproteobacteria. The existence and intracellular location of endobacteria was revealed by fluorescent in situ hybridization (FISH). Our results showed that endobacteria were coccoid, vertically inherited, as yet uncultured, and essential symbionts. Ultrastructural observations indicated that young and old endobacteria differed in cell size, cell wall thickness, and the degree of reproduction. The results of the present study provide a fundamental understanding of the endobacteria inside E. vermicola and raise questions regarding the impact of endobacteria on the biology, ecology, and evolution of their fungal host.

Poplar (Populus spp.) is a high-economic-value tree species around the world. However, poplar trees are damaged by canker disease caused by Valsa sordida. In our previous study, an antagonistic bacterial strain of Streptomyces sioyaensis (No. F57) against V. sordida was isolated, however the antifungal mechanisms by which S. sioyaensis (No. F57) suppress the poplar canker pathogen V. sordida remains unclear. In the present study, S. sioyaensis (No. F57) fermentation broth significantly inhibited mycelial growth. The inhibition rate for mycelial dry weight and colony growth after 3 days was 89.27% and 83.94%, respectively. Spore germination of the pathogen was greatest inhibited 78.30% at 6 h. In vivo biocontrol assays, the disease index of poplar canker treated with S. sioyaensis (No. F57) fermentation broth was significantly reduced compared with the control. At concentrations of 10-fold diluent of fermentation broth, the control effect was up to 78.08%. The fermentation broth also increased mycelia electrical conductivity and the accumulation of malondialdehyde (MDA). The enzyme activities of succinate dehydrogenase (SDH) and Ca2+-Mg2+-ATPase in the tricarboxylic acid cycle (TCA cycle), Hexokinase (HK), Phosphofructokinase (PFK) and pyruvate kinase (PK) in the glycolytic pathway, and Glutathione peroxidase(GSH-Px) were all disrupted in the pathogen mycelia treated with S. sioyaensis (No. F57) fermentation broth. These result revealed that S. sioyaensis (No. F57) inhibited V. sordida by increasing the cell permeability and disrupting the metabolic pathways of the pathogen. Thus, S. sioyaensis (No. F57) can be a potent biocontrol agent for poplar canker disease management in application.

Symbioses have played pivotal roles in biological, ecological, and evolutionary diversification. Symbiotic bacteria affect the biology of hosts in a number of ways. Esteya vermicola, an endoparasitic nematophagous fungus, has high infectivity in the pine wood nematode (PWN), which causes devastating ecological damage and economic losses in Asia and Europe. An integration of molecular, phylogenetic, and morphological analyses revealed that surface-sterilized E. vermicola with septate hyphae from different geographic locations harbor bacterial endosymbionts. 16S rRNA gene sequences from four fungal strains all clustered in a well-supported monophyletic clade that was the most closely related to Pseudomonas stutzeri and affiliated with Gammaproteobacteria. The existence and intracellular location of endobacteria was revealed by fluorescent in situ hybridization (FISH). Our results showed that endobacteria were coccoid, vertically inherited, as yet uncultured, and essential symbionts. Ultrastructural observations indicated that young and old endobacteria differed in cell size, cell wall thickness, and the degree of reproduction. The results of the present study provide a fundamental understanding of the endobacteria inside E. vermicola and raise questions regarding the impact of endobacteria on the biology, ecology, and evolution of their fungal host.

The complete mitochondrial genome of the Nematophagous fungus Esteya vermicola CBS 115803 was determined using the PacBio RS II sequencing technology. The circular molecule is 47,282bp in length with a GC content of 24.85%. Annotated genes including 14 conserved protein-coding genes, the large and the small rRNA subunit (rnl and rns) and 27 tRNAs. The phylogenetic analysis showed that E. vermicola had close genetic relationship with the genus Sporothrix.