Cloning of sft-4 and its influence on vitality and virulence of pine wood nematode, Bursaphelenchus xylophilus

Abstract

In our previous screening of the transcriptome of the causal agent of the devastating pine wilt disease, pine wood nematode (PWN, Bursaphelenchus xylophilus), after treatment with the nematicide fomepizole, Surfeit locus gene sft-4, which encodes a regulatory factor, was found to be downregulated. In situ hybridization results showed that the sft-4 was continuously expressed from egg to adult and was especially high in the reproductive system. Here in a study of the effect of RNA interference (RNAi) of sft-4 and recombinant SFT-4 on PWN activity, treatment with sft-4 dsRNA inhibited feeding, reproduction, oviposition and egg hatching of PWN with the greatest inhibition on reproduction and oviposition, whereas recombinant SFT-4 had the opposite effect. In addition, RNAi of sft-4 changed the female–male ratio and lifespan of PWN. In bioassays of PWNs, with RNAi of sft-4 on seedlings and 2-year-old Pinus thunbergii trees, none of the treated plants developed symptoms during the monitoring period, indicating that virulence of PWNs was either significantly weakened. These results indicate that the influence of sft-4 on PWN pathogenicity may be mainly through regulating reproductive function of PWN and its lifespan.

Keywords

Black pine / Pinaceae / Bursaphelenchus xylophilus / Sft-4 / In situ hybridization / RNAi / Pathogenicity