Abstract
Pine wilt disease (PWD), caused by the pinewood nematode (PWN) Bursaphelenchus xylophilus, is one of the most destructive diseases in trees of the genus Pinus and is responsible for significant environmental and economic losses in North America, Eastern Asia, and Western Europe. However, pine species are not equally affected, with some being tolerant/resistant while others are susceptible to nematode infection for reasons still unclear. The present study aims to investigate differential chemical responses of susceptible and tolerant/resistant pine species shortly after nematode infection by characterizing the phenolic profiles of adult Pinus sylvestris, Pinus pinaster, Pinus pinea, and Pinus halepensis. HPLC and LC-MS were used to identify and quantify the pine´s phenolic compounds: gallic acid, ferulic acid, taxifolin, rutin, resveratrol, (+)-secoisolariciresinol, (−)-epicatechin, protocatechuic acid hexoside, gallic acid hexoside, ferulic acid glucoside, quercetin hexoside, and two unidentified compounds (#A and #B). Prior to infection, we could not differentiate between nematode-tolerant/resistant and susceptible adult pine species based on their constitutive phenolic compounds. In the presence of the PWN, the phenolic profile allowed for a noticeable separation of the PWN-tolerant/resistant P. halepensis from the susceptible P. sylvestris, contrasting with a more homogenous response from P. pinea and P. pinaster. Observations on P. halepensis suggest that taxifolin, resveratrol, and rutin may have an active role in protecting against B. xylophilus, possibly in conjugation with other biochemical and anatomical characters. We emphasize the importance of studying pine tolerant/resistance on adult trees, and not on excised branches, saplings, or seedlings to accurately simulate the nematode–pine host interactions occurring under natural conditions.