Beauveria bassiana strain having insect intestinal infection effect, and application thereof in pest prevention and treatment
A technology of Beauveria bassiana and insects, which is applied in the fields of biotechnology and pesticides, and can solve the problems of restricting field control effects and limiting the large-scale application of fungal pesticides.
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Embodiment 1
[0085] Example 1. Isolation and Identification of Bacterial Strain Bb-bm01
[0086] The bacterial strain Bb-bm01 of the present invention is a new bacterial strain of Beauveria bassiana isolated and purified from the silkworm worm. Bb-bm01 was specifically screened by the following method: under sterile conditions, put the silkworm covered with white hyphae all over the body on the sterilized SDAY medium plate and shake gently, and then place the SDAY plate at 27°C for cultivation Cultivate in an incubator for 3 to 4 days, pick a single colony to a purification medium (SDAY), and carry out purification culture in a 27°C incubator. It was found that a strain similar to Beauveria bassiana had a velveteen-like round colony on SDAY medium, the color gradually changed from white to milky white or light yellow, the hyphae were fluffy, and the powder layer was loose, and it was named Bb-bm01 ( figure 1 ).
[0087] Bb-bm01 was sent to the General Microorganism Center (CGMCC) of C...
Embodiment 2
[0095] Embodiment 2. The method of feeding insect Beauveria bassiana spores orally
[0096] The invention establishes a method for orally feeding the insect Beauveria bassiana. Take the silkworm, a model animal of Lepidoptera, as an example:
[0097] Prepare a certain concentration of fresh Beauveria bassiana spore suspension with 0.01% Triton X-100 sterile water. The silkworm larvae on the second day of the fifth instar were selected. Fix the head of the larva, draw 1 μl of spore suspension with a micropipette (Gilson, P2), and slowly inject it into the mouth of the silkworm larvae ( figure 2 ). The whole process prevents the spore suspension from contacting the mouthparts and body wall of the larvae, and prevents the spores of Beauveria bassiana from infecting insects through the body wall. Then it is placed in an incubator with a temperature of 27°C and a humidity of 30-40%, and is fed with fresh mulberry leaves.
Embodiment 3
[0098] Example 3. Beauveria bassiana Bb-bm01 enteric infection of Lepidoptera larvae
[0099] Inoculate Beauveria bassiana Bb-bm01 on SDAY medium, culture in 27°C incubator for 10 days, then prepare conidia suspension with 0.01% Triton X-100 sterile water, and measure spores with hemocytometer Concentration, respectively dubbed 1×10 9 , 1×10 8 , 1×10 7 , 1×10 6 , 1×10 5, 1×10 4 Conidia / ml concentration of spore suspension.
[0100] A. Targeting the second day larvae of the fifth instar of silkworm P50 strain
[0101] The second-instar larvae of the fifth instar silkworm P50 strain were used to test the intestinal toxicity of Beauveria bassiana Bb-bm01. The feeding conditions of the silkworm larvae were 27°C and 40% humidity. Take 1 μl of different concentrations of Bb-bm01 spore suspension, and use a micropipette to orally feed the second-instar larvae of the fifth instar silkworm P50, and the blank control is 0.01% Triton X-100. There were 30 silkworm larvae in ea...
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