Chengjian Xie

Esteya vermicola is a nematophagous fungus with strong parasitic ability against the pinewood nematode (Bursaphelenchus xylophilus) and shows great potential for the biological control of pine wilt disease. However, this fungus is highly sensitive to environmental stress factors and often exhibits early necrosis when cultured on conventional nutrient-rich media, limiting its large-scale application. In this study, we optimized the long-term cultivation and conidiation conditions of E. vermicola CBS115803 by supplementing minimal medium (MM) with amino acids, and evaluated its stress tolerance and infectivity against the pinewood nematode. Among 20 tested amino acids, histidine significantly increased total conidia production, while arginine, glutamine, and proline markedly promoted the formation of lunate conidia. The combination of arginine, histidine, glutamine, and proline (AHGP) produced the highest overall conidia yield and lunate conidia proportion. The MM + AHGP medium maintained long-term colony viability, whereas colonies on PDA and CM media showed obvious degeneration. This formulation also improved mycelial growth, total conidiation, and the proportion of lunate conidia. Moreover, conidia produced on MM + AHGP exhibited the highest germination rates and infectivity under various stress conditions, including cold, heat, oxidative, osmotic, and UV stresses. Conidia germination was significantly enhanced following treatment at 0 °C, suggesting that low temperatures may activate dormancy-breaking pathways. This amino acid-optimized medium offers an effective technical foundation for stable large-scale production and storage of E. vermicola conidia, providing a new avenue for the biocontrol of pine wilt disease.

Chrysanthemum morifolium Ramat. is a major ornamental crop that suffers from diverse fungal, bacterial, viral, and insect pests, causing significant yield and quality losses. Between 2015 and 2025, rapid progress in molecular biology, genomics, and ecological regulation has advanced both fundamental research and applied control strategies. Multi-locus sequencing, multiplex PCR, and next-generation sequencing refined the identification of fungal and bacterial pathogens, while functional studies of WRKY, MYB, and NAC transcription factors revealed key resistance modules. Hormone-mediated signaling pathways, particularly those of salicylic acid, jasmonic acid, and abscisic acid, were shown to play central roles in host defense. Despite these advances, durable genetic resistance against bacterial pathogens and broad-spectrum defense against viruses remains limited. Novel technologies, including virus-free propagation, RNA interference, and spray-induced gene silencing, have shown promising outcomes. For insect pests, studies clarified the damage and virus-vectoring roles of aphids and thrips, and resistance traits linked to trichomes, terpenoids, and lignin have been identified. Biocontrol agents such as Trichoderma spp., Bacillus spp., predatory mites, and entomopathogenic fungi have also demonstrated efficacy. Future efforts should integrate molecular breeding, genome editing, RNA-based tools, and microbiome management to achieve sustainable chrysanthemum protection.

BACKGROUND: Sapindus mukorossi , a versatile woody plant utilized in biomass energy, biotechnology and landscaping, has been the focus of this study. The primary objective was to assess the antifungal potential of soapberry pericarp extracts against several common plant pathogenic fungi. RESULTS: The investigation revealed that the soapberry pericarp extract displayed broad‐spectrum antifungal activity against six plant pathogenic fungi. Notably, it effectively inhibited the spore germination of Verticillium dahliae and Fusarium oxysporum . In subsequent inoculation trials with V. dahliae , the soapberry extract significantly reduced the incidence of cotton verticillium wilt compared to the control. Additionally, cotton treated with the extract showed a higher upregulation of disease‐resistance genes PR1 and PR5 postinfection with V. dahliae . The extract also suggested a potential growth‐promoting effect. In fruit biocontrol assays, it demonstrated remarkable antifungal activity against Botryitis cinerea in tomato, preventing mycelial expansion at the inoculation site, and reduced Penicillium digitatum infection in citrus to merely 13%, in contrast to 85% in the control. Extraction experiments further confirmed that using water as the solvent was the most efficient and economical method for extracting antifungal active components from soapberry pericarp. CONCLUSION: The study concluded that soapberry pericarp extracts possess significant broad‐spectrum antifungal properties, showing promise as a potential alternative for plant disease management and fruit biocontrol. The water extraction method identified offers an effective and cost‐effective way to harness these beneficial antifungal components. © 2025 Society of Chemical Industry.

Bursaphelenchus xylophilus, the causative agent of pine wilt disease (PWD), poses a severe global threat to coniferous forests. Esteya vermicola, an endoparasitic nematophagous fungus, exhibits promising biocontrol potential against this pinewood nematode. The vesicular transport system, evolutionarily conserved in eukaryotes, is essential for fungal pathogenicity. Based on our genome sequence of E. vermicola CBS115803, we identified EvSec22, a gene encoding a SNARE protein implicated in vesicular transport process. This study investigates the role of EvSec22 in E. vermicola during nematode infection, utilizing our optimized gene knockout methodology. Infection assays revealed that EvSec22 deletion significantly impaired the pathogenicity of E. vermicola against B. xylophilus. Phenotypic analyses revealed that the ΔEvSec22 mutant exhibited suppressed hyphal growth, reduced conidiation, and abnormal septal spacing. Furthermore, the mutant showed significantly diminished tolerance to osmotic stress (sorbitol) and oxidative stress (hydrogen peroxide). Overall, the EvSec22 gene is associated with the virulence of E. vermicola CBS115803 against B. xylophilus, and its deletion also affects the normal growth of E. vermicola and its tolerance to abiotic stress. This study providing new insights into SNARE protein functions in fungal biocontrol agents.

Bursaphelenchus xylophilus , the causative agent of pine wilt disease (PWD), poses a severe global threat to coniferous forests. Esteya vermicola , an endoparasitic nematophagous fungus, exhibits promising biocontrol potential against this pinewood nematode. The vesicular transport system, evolutionarily conserved in eukaryotes, is essential for fungal pathogenicity. Based on our genome sequence of E. vermicola CBS115803, we identified EvSec22, a gene encoding a SNARE protein implicated in vesicular transport process. This study investigates the role of EvSec22 in E. vermicola during nematode infection, utilizing our optimized gene knockout methodology. Infection assays revealed that EvSec22 deletion significantly impaired the pathogenicity of E. vermicola against B. xylophilus. Phenotypic analyses revealed that the ΔEvSec22 mutant exhibited suppressed hyphal growth, reduced conidiation, and abnormal septal spacing. Furthermore, the mutant showed significantly diminished tolerance to osmotic stress (sorbitol) and oxidative stress (hydrogen peroxide). Overall, the EvSec22 gene is associated with the virulence of E. vermicola CBS115803 against B. xylophilus, and its deletion also impacts the normal growth of E. vermicola and its tolerance to abiotic stress. This study providing new insights into SNARE protein functions in fungal biocontrol agents.

Verticillium dahliae, a virulent soil-borne fungus, elicits Verticillium wilt in numerous dicotyledonous plants through intricate pathogenic mechanisms. Ubiquitination, an evolutionarily conserved post-translational modification, marks and labels proteins for degradation, thereby maintaining cellular homeostasis. Within the ubiquitination cascade, ubiquitin ligase E3 demonstrates a unique capability for target protein recognition, a function often implicated in phytopathogenic virulence. Our research indicates that two ubiquitin ligase E3s, VdBre1 and VdHrd1, are intrinsically associated with virulence. Our findings demonstrate that the deletion of these two genes significantly impairs the ability of V. dahliae to colonize the vascular bundles of plants and to form typical penetration pegs. Furthermore, transcriptomic analysis suggests that VdBre1 governs the lipid metabolism pathway, while VdHrd1 participates in endoplasmic-reticulum-related processes. Western blot analyses reveal a significant decrease in histone ubiquitination and histone H3K4 trimethylation levels in the ΔVdBre1 mutant. This research illuminates the function of ubiquitin ligase E3 in V. dahliae and offers fresh theoretical perspectives. Our research identifies two novel virulence-related genes and partially explicates their roles in virulence-associated structures and gene regulatory pathways. These findings augment our understanding of the molecular mechanisms inherent to V. dahliae.

BACKGROUND: The pinewood nematode ( Bursaphelenchus xylophilus ) causes severe damage to pine trees. The nematophagous fungus, Esteya vermicola , exhibits considerable promise in the biological control of Bursaphelenchus xylophilus due to its infectivity. Notably, the lunate conidia produced by E. vermicola can infect Bursaphelenchus xylophilus . In the study, we aim to investigate the genes involved in the formation of the lunate conidia of E. vermicola CBS115803. RESULTS: Esteya vermicola CBS115803 yielded 95% lunate conidia on the complete medium (CM) and 86% bacilloid conidia on the minimal medium (MM). Transcriptomic analysis of conidia from both media revealed a significant enrichment of differentially expressed genes in the pathway related to ‘cellular amino acid biosynthesis and metabolism’. Functional assessment showed that the knockout of two arginine biosynthesis genes ( EV232 and EV289 ) resulted in defects in conidia germination, mycelial growth, lunate conidia formation, and virulence of E. vermicola CBS115803 in Bursaphelenchus xylophilus . Remarkably, the addition of arginine to the MM improved mycelial growth, conidiation and lunate conidia formation in the mutants and notably increased conidia yield and the lunate conidia ratio in the wild‐type E. vermicola CBS115803. CONCLUSION: This investigation confirms the essential role of two arginine biosynthesis genes in lunate conidia formation in E. vermicola CBS115803. The findings also suggest that the supplementation of arginine to the culture medium can enhance the lunate conidia yield. These insights contribute significantly to the application of E. vermicola CBS115803 in managing Bursaphelenchus xylophilus infections. © 2023 Society of Chemical Industry.

Pine wilt disease, which is caused by the nematode Bursaphelenchus xylophilus, is one of the most destructive forest diseases worldwide. Esteya vermicola, a nematophagous fungus, has emerged as a promising biological control agent. However, the limited availability of gene function analysis techniques hinders further genetic modification of this fungus. In this study, we employed a combination of enzymes (driselase, snailase, and cellulase) to enzymatically degrade the cell wall of the fungus, resulting in a high yield of protoplasts. Furthermore, by utilizing 0.6 M sucrose as an osmotic pressure stabilizer, we achieved a significant protoplast regeneration rate of approximately 31%. Subsequently, we employed the polyethylene glycol-mediated protoplast transformation method to successfully establish a genetic transformation technique for E. vermicola CBS115803. Additionally, through our investigation, we identified the Olic promoter from Aspergillus nidulans, which effectively enhanced the expression of the DsRed gene encoding a red fluorescent protein in E. vermicola CBS115803. Moreover, we successfully implemented a split-marker strategy to delete the EvIPMD gene in E. vermicola CBS115803. In summary, our findings present valuable experimental methodologies for gene function analysis in E. vermicola CBS115803.

Mahonia fortunei, belonging to the Berberidaceae family, is widely cultivated in fields, parks, courtyards, and roadsides for its excellent ornamental characteristics and medicinal values in southe...