Separation and purification method of beauveria bassiana
A technology for separation and purification of Beauveria bassiana, applied in the field of application, can solve the problems of limited practical application, high application cost, carcinogenicity of cycloheximide, etc., and achieve good practical application prospects, simple method, and cost reduction effect
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Embodiment 1
[0043] Treat the collected pathogens: use 75% (v / v) ethanol cotton balls to disinfect the surface of the silkworm body of the naturally infected Ambrysalis larvae (which can also be artificially infected by conventional means), and then put it into the sterile Put a group of cotton balls soaked with sterile water in the dish, and culture it at 26°C for 3-5 days (d). After the silkworm body surface grows white fluffy hyphae and conidia For strain isolation;
[0044] S1. Preparation of spore liquid: use a sterile inoculation loop to pick 2 to 3 rings of conidia powder from the surface of the silkworm body, put them in a triangular flask filled with 50 mL of sterile water (containing sterile glass beads), and vortex for 5 min, to fully disperse the spores;
[0045] S2. Prepare the plate:
[0046] S21. Prepare the following reagents: the concentration of streptomycin sulfate is 1000U / mL (prepared with sterile water, without autoclaving, and added before use); the mass percentage...
Embodiment 2
[0053] Treat the collected pathogens: sterilize the surface of silkworm chrysalis infected with natural infection or artificial infection of Beauveria bassiana with 75% (v / v) ethanol cotton balls, then put it into a sterile petri dish, and put it in the dish for a period of time. Group cotton balls soaked in sterile water, place them at 26°C and incubate them for 3-5 days (d), and isolate the strains after the white fluff mycelia and conidia grow on the surface of the silkworm body;
[0054] S1. Preparation of spore liquid: use a sterile inoculation loop to pick 2 to 3 rings of conidia powder from the surface of the silkworm body, put them in a triangular flask filled with 50 mL of sterile water (containing sterile glass beads), and vortex for 5 min, to fully disperse the spores;
[0055] S2. Prepare the plate:
[0056] S21. Prepare the following reagents: the concentration of streptomycin sulfate is 10000U / mL (prepared with sterile water, without autoclaving, and added befor...
Embodiment 3
[0063] S1. Dilute the biopesticide containing Beauveria bassiana spores for biological control (randomly selected by Guangzhou Biological Control Station, those skilled in the art can also use similar biopesticides containing Beauveria bassiana spores for experiments) to a mass Dilutions with a percentage concentration of 1 or 10%.
[0064] S2. preparing a plate;
[0065]S21. Prepare the following reagents: the concentration of streptomycin sulfate is 100000U / mL (prepared with sterile water, without autoclaving, and added before use); the mass percentage concentration of sodium deoxycholate is 2.2%, and the mass percentage of Bengal red Concentration is 0.25%, autoclave for standby;
[0066] S22. Peel 200g of potatoes, cut them into small pieces, add 1000mL of water to boil until rotten (boil for 20-30min, it can be pierced with a glass rod), filter with 4 layers of gauze, add 20g of agar and 20g of glucose Afterwards, add water to 1000 mL, adjust the pH of the medium to 6.0...
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