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.
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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