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.
To clarify the role of bacteria in PWD, three PWNs with different virulence (strongly virulent strain AMA3, normally virulent strain AA3, and weakly virulent strain YW4) were selected as research objects, and three strains of Bacillus cereus (nematode-associated bacteria GD1, Pinus massoniana endophytic bacteria GD2, and P. elliottii endophytic bacteria NJSZ-13) at different concentrations were used to determine their effects on the survival and fecundity of the nematodes. The results showed that strains GD1 and GD2 could significantly improve the survival and fecundity of PWNs at three different concentrations, while NJSZ-13 showed the opposite effects. The inoculation experiments showed that the disease index of P. massoniana under different treatments was as follows: AMA3 < a mixture of AMA3 and GD1 < a mixture of AMA3 and GD2. Similar results were shown in the Larix kaempferi inoculation experiment. Further, using RNA-sequencing analysis, we found that the up-regulated genes in PWN were sHsp 21, Hsp 70, and Hsp 72 after being treated by strains GD1 and GD2. The longevity regulatory pathways, MAPK signaling pathways, glutathione metabolic pathways, and cytochrome P450 metabolic pathways related to these genes are clearly enriched. These results show that the bacteria can improve the host adaptability of PWN, and endophytic bacteria of pine trees may be more effective in improving the host adaptability of nematodes than the associated bacteria of nematodes.
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