In previous research, the pine wood nematode Bxadh2 gene significantly increased its expression in pine seedlings inoculated with endophytic Bacillus cereus GD2 and pine wood nematode. To explore pine wood nematode Bxadh2 gene function, we cloned and analyzed its biological information, and we preliminarily studied its physiological function through RNA interference. We found that the Bxadh2 gene’s full CDS length is 1269 bp, which encodes 422 amino acids, and presents a relatively stable hydrophobic protein. The protein encoded by the Bxadh2 gene has no signal peptide or transmembrane structure, and it is an intracellular protein that does not participate in transmembrane movement. The RNAi interference results showed that when the pine wood nematode’s Bxadh2 gene was suppressed, its survival rate and fecundity significantly decreased, indicating that the expression of the Bxadh2 gene was related to the growth and development of pine wood nematodes.

Pine wood nematode disease is a destructive disease to the ecological environment and forest resources. The disease is produced by Bursaphelenchus xylophilus, and the onset rate is extremely fast and the pathogenesis of the disease is not clear. Prevention of this disease is still important in production, so it is of great significance to explore its early diagnosis. In this study, the changes of chlorophyll fluorescence dynamic induction parameters, non-photochemical quenching parameters and organic acid content in needles of 7-year-old-infected Pinus hwangshanensis were studied by chlorophyll fluorescence technology and organic acid mass combined technology. The results showed that in the chlorophyll fluorescence dynamic induction group, Fm/Fo, Fv/Fo, Fm, Psi_o and Pi_Abs showed a significant downward trend, while Phi_Do, ABS/RC, TRO/RC and DIO/RC showed a significant uptrend. In non-photochemical quenching, Fm showed a downward trend, while Qp_L1, Qp_L2, QY_L1, QY_L2 and QY_Lss showed a significant upward trend. In the change of organic acid content, ferulic acid, syringic acid, gallic acid and jasmonic acid showed a significant increasing trend, while benzoic acid and salicylic acid showed a first increasing trend and then decreasing trend. Experimental results show that pine wood nematode had significant effects on photosynthesis and the organic acid content of pine before it showed symptoms of the disease. After comprehensive analysis, chlorophyll fluorescence dynamic induction parameters Pi_Abs, Fm, Phi_Do, TRO/RC, Psi_o, non-photochemical quenching parameters QY_L2, Qp_L1, QY_Lss, QY_L1 and organic acids ferulic acid, syringic acid, benzoic acid and gallic acid contents are ideal indicators for early diagnosis of pine wood nematodiasis.

To explore the effect of associated bacteria on the low-temperature adaptability of pinewood nematodes (PWNs), transcriptome sequencing (RNA-seq) of PWN AH23 treated with the associated bacterial strain Bacillus cereus GD1 was carried out with reference to the whole PWN genome. Bioinformatic software was utilized to analyze the differentially expressed genes (DEGs). This study was based on the analysis of DEGs to verify the function of daf-11 by RNAi. The results showed that there were 439 DEGs between AH23 treated with GD1 and those treated with ddH2O at 10 °C. There were 207 pathways annotated in the KEGG database and 48 terms annotated in the GO database. It was found that after RNAi of daf-11, the survival rate of PWNs decreased significantly at 10 °C, and fecundity decreased significantly at 15 °C. It can be concluded that the associated bacteria GD1 can enhance the expression of genes related to PWN low-temperature adaptation and improve their adaptability to low temperatures.