The invention discloses a method for preparing
bacteria-free
pine wood nematodes. The method for preparing the
bacteria-free
pine wood nematodes comprises the following steps: culturing the
pine wood nematodes; preparing pure pine wood
nematode eggs; soaking the pine wood
nematode eggs in 15% H2O2 for 50-60 minutes; incubating the pine wood nematodes; and culturing the pine wood nematodes and performing
bacteria-free identification, thereby preparing the bacteria-free pine wood nematodes. As for the method for preparing the bacteria-free pine wood nematodes, the property of being capable of being adhered on a
glass vessel of the pine wood
nematode eggs is utilized, a small culture dish is adopted for collecting the eggs, the newly prepared 15% H2O2 is further used for performing sterilization treatment, the bacteria-free pine wood nematode eggs are obtained, the bacteria-free eggs are incubated,
mass propagation is further performed on
botrytis cinerea, and then the bacteria-free pine wood nematodes are finally obtained. Under sterile conditions, the obtained bacteria-free pine wood nematodes are used for performing an
inoculation test on seedlings cultured by spruce bacteria-free tissues, and the result shows that the
inoculation of the bacteria-free pine wood nematodes can still enable the bacterial-free seedlings to wilt. The method can be adopted for enabling the pine wood nematodes to be bacteria-free and lose
pathogenicity, thereby being the simple and successful method.