Verticillium lecanii for preventing and controlling fly type pests and use thereof
A technology for Verticillium lecanii and pests, applied in the fields of application, insecticides, fungi, etc., to achieve the effects of simple cultivation, good pathogenicity of adults, and rapid growth
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Embodiment 1
[0007] Embodiment one, the isolation of pathogenic bacteria, identification
[0008] 1.1 Materials and methods
[0009] 1.1.1 Materials
[0010] The adults of Boettcheriscaperegrine (Diptera: Sarcophagidae) infected by an entomogenic fungus were collected in Kunming.
[0011] Potato dextrose agar medium (PDA): 200g potato + 17-20g agar + 17-20g glucose + 1000ml water.
[0012] Sterile operating conditions: All utensils and utensils are sterilized in a high-temperature autoclave (121°C, 30min), and inoculation and other operations are performed in an ultra-clean workbench.
[0013] Culture conditions: Culture in a 29°C light (12L:12D) incubator. After the colonies are formed, transfer to the test tube PDA slope, culture for 2 to 3 days, and transfer to a 4°C refrigerator for storage.
[0014] 1.1.2 Isolation and purification of pathogenic bacteria
[0015] Isolation Pathogens were isolated from the carcasses of diseased R. Soak the dead body of the adult sarcophagus in seq...
Embodiment 2
[0024] Embodiment two, the biological characteristics of the purified strain of Verticillium lecanii
[0025] 2.1 Materials and methods
[0026] 2.1.1 The strains to be tested Select a plate that grows uniformly and vigorously after purification as the strains to be tested. The hyphae were inoculated on PDA again, and cultivated in a constant temperature light (12L:12D) incubator at 29°C.
[0027] 2.1.2 Determination of colony growth rate and sporulation amount Take a plate of Verticillium lecanii cultivated in advance and punch holes with an 8mm hole puncher, inoculate it on a new medium, do three repetitions, and place at 29 Cultivate in a constant temperature and light (12D:12L) incubator at ℃, measure and record its diameter regularly every day, until the colony is overgrown with the culture medium. Use a hole puncher with a diameter of 8mm to take the bacterium cake at the same position of the culture medium, add 1% Tween-80 and 20ml sterile water, wash the spores to ma...
Embodiment 3
[0037] Example 3. Indoor pathogenicity determination of purified strains of Verticillium lecanii to Sarcophagus spp., Lucilia sericata, Musca domestica, Tysofly, and Drosophila
[0038] 3.1 Materials and methods
[0039] 3.1.1 Source of tested insects
[0040]Select the same batch of 4-day-old, disease-free, and uniformly sized adult larvae, Lucilia sericata, housefly, casefly, and fruit fly as test insects, and put the test insects separately into small insect cages , fed with water and white sugar, and reared under the conditions of 29±1°C and a photoperiod of 12L:12D.
[0041] 3.1.2 Preparation of spore suspension
[0042] Take the well-growing purified Verticillium lecanii and wash the spores with sterile water and filter (generally, the number of spores in the filtrate can reach 10 8 spores / ml), use a sterile capillary pipette to take a drop of the filtrate and drop it on a hemocytometer, count the spores under a microscope and record the data, and then dilute to 10 wi...
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