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<p>Pine wilt disease (PWD) is a major threat to pine forests worldwide and is caused by the plant-parasitic nematode <i>Bursaphelenchus xylophilus</i>. Despite its ecological and economic impact, the miRNA-mediated regulatory landscape of this pathogen remains poorly characterized. Here, we performed an integrative computational analysis to identify putative miRNAs in <i>B. xylophilus</i> and to prioritize candidate regulatory targets potentially relevant to pathogenicity. Using expressed sequence tag (EST) data and a stepwise screening pipeline, we predicted 22 putative novel miRNAs whose precursor sequences formed canonical stem–loop hairpin structures with consistently negative minimum free energy values (mean − 26.73&#xa0;kcal/mol), supporting thermodynamically stable folding. Phylogenetic analysis grouped these miRNAs into three major clusters. To facilitate downstream functional interpretation, we additionally performed a supporting protein annotation update and curated a set of 14 PWD-associated proteins from the literature. Predicted miRNA–target analysis and network construction suggested that bxy-miR-4731-5p and bxy-miR-607 may convergently target USP15, a ubiquitin-specific protease family gene implicated in nematode development and parasite–host interaction processes. Overall, this study provides a resource of putative miRNAs and prioritized candidate regulatory modules in <i>B. xylophilus</i> and proposes testable hypotheses for future experimental validation, which may ultimately support the development of improved control strategies for PWD.</p>

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Identification of putative novel miRNAs and predicted regulatory targets in Bursaphelenchus xylophilus, the causal agent of pine wilt disease

  • 期刊论文

  • 发表日期：

    2026年5月8日

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  • Caoli Zhu,
  • Yicheng Yan,
  • Jiawei Sun,
  • Yongheng Wang,
  • Zhiyuan Yang

摘要

Pine wilt disease (PWD) is a major threat to pine forests worldwide and is caused by the plant-parasitic nematode Bursaphelenchus xylophilus. Despite its ecological and economic impact, the miRNA-mediated regulatory landscape of this pathogen remains poorly characterized. Here, we performed an integrative computational analysis to identify putative miRNAs in B. xylophilus and to prioritize candidate regulatory targets potentially relevant to pathogenicity. Using expressed sequence tag (EST) data and a stepwise screening pipeline, we predicted 22 putative novel miRNAs whose precursor sequences formed canonical stem–loop hairpin structures with consistently negative minimum free energy values (mean − 26.73 kcal/mol), supporting thermodynamically stable folding. Phylogenetic analysis grouped these miRNAs into three major clusters. To facilitate downstream functional interpretation, we additionally performed a supporting protein annotation update and curated a set of 14 PWD-associated proteins from the literature. Predicted miRNA–target analysis and network construction suggested that bxy-miR-4731-5p and bxy-miR-607 may convergently target USP15, a ubiquitin-specific protease family gene implicated in nematode development and parasite–host interaction processes. Overall, this study provides a resource of putative miRNAs and prioritized candidate regulatory modules in B. xylophilus and proposes testable hypotheses for future experimental validation, which may ultimately support the development of improved control strategies for PWD.