Bursaphelenchus xylophilus (pine wood nematode, PWN) has been present in China for over 40 years and has spread to northeast China, where native pine species are key components of the local top community. Pinus thunbergii is known to be susceptible to PWN among local conifer species, whereas research on PWN’s pathogenicity in Larix remains limited. Furthermore, there are no research reports on PWN infestation in Picea and Abies species within China. This study conducted a detailed analysis of phenotypic changes and temporal spectral reflectance variations in four conifer species in northeast China—P. thunbergii, Larix kaempferi, Picea koraiensis, and Abies holophylla—following artificial inoculation with PWN. The aim of this study is to establish a theoretical basis for identifying the potential hosts and threats of PWN. The study incorporated a 60-day post-inoculation observation period to systematically monitor and compare temporal changes in external morphology, disease susceptibility (incidence and mortality rates), spectral reflectance, and the normalized wilt index (NWI) in 2–3-year-old seedlings of P. thunbergii, L. kaempferi, P. koraiensis, and A. holophylla after inoculation with PWN. The results showed that P. thunbergii displayed the earliest infection symptoms, followed by L. kaempferi, A. holophylla, and finally P. koraiensis. After inoculation, P. thunbergii was the first to experience mortality, followed by L. kaempferi, P. koraiensis, and A. holophylla. Following inoculation, P. thunbergii exhibited the earliest significant increase in NWI (p < 0.001), followed by L. kaempferi and A. holophylla; P. koraiensis showed the latest increase (p < 0.001). In conclusion, the experiment identified P. koraiensis as having the strongest resistance to PWN among the four species, followed by A. holophylla. P. thunbergii showed the weakest resistance, while L. kaempferi exhibited moderate resistance. The ranking of PWN susceptibility for the four conifer species, from highest to lowest, is as follows: P. thunbergii, L. kaempferi, A. holophylla, and P. koraiensis.

The pine wood nematode (Bursaphelenchus xylophilus, PWN) is a globally significant quarantine pest that causes severe economic and ecological damage to coniferous forests worldwide. Additionally, PWNs continue to expand into higher latitudes. However, studies on their cold tolerance remain limited. This study investigated the overwintering environment of PWNs in epidemic areas of Liaoning Province, China. It established a protocol to induce anhydrobiosis in PWNs, evaluated their low-temperature resistance, observed morphological changes during anhydrobiosis, and explored potentially involved key genes. The results showed that (1) there was no significant difference in thermal insulation between infected and healthy wood in Liaoning Province; both effectively reduced temperature fluctuation rates, providing a protective function for PWN overwintering. (2) PWNs significantly enabled their cold tolerance through anhydrobiosis, accompanied by significant morphological changes and substantial lipid droplet depletion. (3) Eleven anhydrobiosis-related genes were identified. Among these, the collagen gene family showed consistent expression patterns throughout dehydration and rehydration. This suggests a potential role in cuticle structural changes and osmoregulation during anhydrobiosis. These findings provide a theoretical basis for understanding how PWNs survive winter conditions in high-latitude regions. Additionally, they offer valuable insights for future research into PWN anhydrobiosis and the development of effective control strategies.