Method for preparing bacteria-free pine wood nematodes

Abstract

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.

Description

Technical field [0001] The invention relates to bacteria-free pine wood nematodes, in particular to a method for preparing bacteria-free pine wood nematodes. Background technique [0002] Pine wood nematode disease [Bursaphelenchu ​​xylophilus (Sleiner&Buhrer) Nickle], also known as pine wilt disease, is a devastating disease of pine trees. In view of the danger of the disease, scholars from various countries have conducted systematic and in-depth studies on it, but the pathogenic mechanism has not yet been fully understood. Regarding the pathogen of pine wood nematode disease, there are currently two views. One view is that pine wood nematode is the only pathogen, and the other view is that pine wood nematode disease is a compound disease involving pine wood nematode and its accompanying bacteria. Bacteria play a leading role in science. [0003] Kiyohara and Tokushige (1971) inoculated 14-25 year-old black pine and red pine with pine wood nematodes cultured on polychaete in di...

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Problems solved by technology

Zhao Boguang et al. have also invented a method of obtaining sterile nematodes by surface sterilization of eggs, and Ben Ailing et al. have used this method to cultivate sterile nematodes, but the operation steps are also relatively cumbersome, especially for the picking of eggs. Extremely inconvenient, making it less operable (Ben Ailing et al., 2008)

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