Viability and pathogenicity of Esteya vermicola in pine trees

Esteya vermicola, as the first reported endoparasitic fungus of pinewood nematode (PWN), exhibited high infectivity in vitro and has been patented based on its potential as a bio-control agent against PWN. The isolation substrates and taxonomic status suggested E. vermicola is associated with beetles, saprotrophic and kills nematodes in trees. However, the direct experimental evidence for this was still lacking. In the present studies, beta-tubulin gene was applied to confirm the taxonomic identification of E. vermicola. Furthermore, our results showed that E. vermicola survived resin and other chemicals secreted by pine trees, and reproduced with new lunate conidia to parasitize other migratory PWNs. In order to confirm the pathogenicity of E. vermicola, pine seedlings and large pine trees were inoculated with 300 mu L and 40 mL conidia suspensions (10(9) mL(-1)). The results showed that all treated pine trees were healthy with no differences compared to the controls. Furthermore, necrosis or discoloration caused by this fungus was not observed on wood slices. Basal knowledge was provided for the application of E. vermicola to control PWN in vivo.