Autophagy contributes to the feeding, reproduction, and mobility of Bursaphelenchus xylophilus at low temperatures.

The pine wood nematode (PWN), Bursaphelenchus xylophilus, is the pathogen causing pine wilt disease (PWD), which is a devastating forest disease. At present, little is known about the defense mechanisms of the PWN, which limits PWD control. Although autophagy plays an important role in the physiological and pathological processes of eukaryotes, its significance in the PWN remains unknown. In this study, we prepared an anti-BxATG8 polyclonal antibody and identified two PWN autophagy marker proteins: BxATG8-I and BxATG8-II. By western blot analysis, we found that the ratio of BxATG8-II to BxATG8-I, which represents autophagic activity, was decreased significantly when samples were treated with the autophagy inhibitor 3-methyladenine. As such, we were able to successfully detect and quantify autophagic activity in the PWN. Thereafter, we investigated the effects of low and high temperatures on PWN growth and reproduction. The results revealed that feeding rate, reproduction rate, and mobility decreased at 15°C and increased at 35°C. By contrast, autophagic activity was high at 15°C and low at 35°C, suggesting that the PWN regulates autophagic activity in response to changes in temperature to maintain physiological homeostasis. When autophagy was inhibited at 15°C, feeding rate, reproductive rate, and mobility declined further, indicating that autophagy is crucial for PWN growth and reproduction at low temperature. These results indicate that autophagy in the PWN is an important response mechanism to temperature changes.