Abstract

Simple SummaryThe pine wood nematode (PWN),Bursaphelenchus xylophilus, causes severe damage to pine forests worldwide, yet the molecular mechanisms controlling its gene activity remain poorly understood. In this study, we investigatedN6-methyladenosine (m6A), an RNA common chemical modification that functions as a regulatory switch controlling gene expression. We identified 21 genes inB. xylophilusthat are involved in adding, removing, or recognizing this modification. These genes showed distinct expression patterns across different developmental stages, from eggs to adults. In addition, they also responded significantly to environmental challenges, including low temperature, exposure to host defense compounds, and infection of pine seedlings. Together, these findings provide fundamental insights into m6A–mediated regulation in PWNs and establish a basis for future studies on pine wilt disease.m6A represents a prevalent epitranscriptomic modification in eukaryotes. The dynamic balance of m6A modification is governed by methyltransferases (writers), demethylases (erasers), and binding proteins (readers). m6A regulators are integral to critical biological processes, including embryonic development, cell differentiation, and stress responses.B. xylophilus, a highly destructive invasive plant-parasitic nematode, has caused considerable ecological and economic damage worldwide. However, the m6A regulatory system in PWNs has not yet been investigated. In this study, we systematically identified 21 m6A regulators in PWNs, including 10 writers, 6 erasers, and 5 readers, which belong to the METTL, ALKBH, and KH/RRM families. Phylogenetic and domain analyses revealed the evolutionary conservation and functional diversification of these protein families. Expression profiling indicated stage-specific expression patterns of m6A regulators during the egg, larval, diapause, and adult stages. Furthermore, significant responses were observed under low-temperature treatment, β-pinen