Insecticidal fungus and application thereof
A technology of fungi and insecticides, applied in the direction of application, insecticides, fungi, etc., can solve the problems of lagging aphid reproduction speed tolerance, inability to control insect pests in time, slow onset of host effects, etc., to ensure the safety of agricultural products, Effects of protecting crops and reducing the amount of application
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Embodiment 1
[0036] Example 1 Isolation and purification of Beauveria bassiana BBNS-J9-16 (Beauveria bassiana) bacterial strain
[0037] Soil and corpses infected with Beauveria bassiana were collected from natural forests, grasslands or farmland in Inner Mongolia, Anhui, Jiangxi, Guizhou and other places, and the microorganisms were isolated and purified in the laboratory.
[0038] The separation and purification steps are: 1) Isolate the target microorganism from the soil: dilute the soil 20 times, 100 times, and 500 times with sterile water, inoculate them on selective medium respectively, cultivate them at 25°C for 4-7 days, pick out Transfer the colony to a new selective medium plate, culture a single colony for 7-10 days, and identify Beauveria bassiana by colony appearance and microscopic morphology inspection, transfer to the culture medium test tube, at 25 ℃ After culturing at high temperature for 14 days, they were stored in a refrigerator at 4°C. 2) Isolate target microorganism...
Embodiment 2
[0040] Biomorphological characteristics of the strain of embodiment 2
[0041] Prepare PDAY medium, its components and preparation method are to boil 200g of fresh potatoes and about 1000mL of water for 15min, filter with gauze and retain the liquid, add 20g of glucose, 5g of yeast extract powder, and 15g of agar, heat to completely melt the agar, and quantify to 1000mL, divided into Erlenmeyer flasks, placed in a high-pressure steam sterilizer at 121°C for 20min, and when cooled to about 60°C, pour it into a petri dish to make a plate for use.
[0042] Inoculate conidia on PDAY plate medium, culture at 25°C for 2 days, colonies can be seen with the naked eye, and the mycelia of the colonies are white and fluffy. Continue to cultivate for 3-4 days, and it can be seen that the colonies expand and grow, and milky white spores are produced on the surface. After the spores mature, White powdery spores were enriched on the surface of the colony.
[0043] Drop a drop of sterilized ...
Embodiment 3
[0044] Ribosome ITS region nucleotide sequence characteristics of embodiment 3 bacterial strains
[0045] Prepare LM medium, its components and preparation method are as follows: weigh 40g of glucose and 5g of peptone, add water to 1000mL, divide into Erlenmeyer flasks, place in a high-pressure steam sterilizer at 121°C for 20min, and let cool to room temperature for later use.
[0046] Inoculate the conidia into the above-mentioned medium, the inoculum size is 1×10 5 Spores / mL, cultured on a shaker at 25±1°C and 200rpm for 48h, filtered through filter paper, retained mycelia, squeezed dry for later use.
[0047] Genomic DNA was extracted by CTAB method. According to the routine DNA extraction operation, the genomic DNA is obtained, such as figure 2 shown.
[0048] ITS1-5.8S-ITS2 rDNA region primer 15'-TCCGTAGGTGAACCTGCGG and primer 25'-TCCTCCGCTTATTGATATGC were used for PCR amplification to obtain amplified DNA fragments, such as image 3 As shown, the fragment was subje...
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