A kind of biological source insecticide and preparation method thereof
An insecticide and biological source technology, applied in the field of biological control, can solve the problems of lack of high pathogenicity and achieve the effect of strong toxicity, increased cost and thorough effect
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Embodiment 1
[0028] Embodiment 1: obtain pathogenic fungus
[0029] The brown planthopper carcasses were collected from rice fields in the agricultural area of Yuyao City, Zhejiang Province, and stored in a 4°C refrigerator for later use. The population of the tested brown planthopper was established and preserved in the laboratory. It was reared in isolation with TN1 rice seedlings in an artificial climate room (temperature 26°C±1°C, photoperiod 14L:10D, relative humidity 75%-85%) to maintain the population; TN1 rice planting The conditions were the same as those of brown planthopper, and they were used in the experiment after the tillering stage.
[0030] Isolation and purification of pathogenic fungi: Potato dextrose agar medium (PDA) was used as the separation and purification medium. First, the surface of the dead body of the brown planthopper was sterilized with 100% alcohol, washed 5 times with sterile distilled water, dried and pounded. Broken insect body. Appropriately dilute ...
Embodiment 2
[0032] Embodiment 2: identification and preservation of pathogenic fungi
[0033] Morphological identification: On the PDA medium, the colony was initially white and filamentous, and then gradually expanded, reaching an average diameter of 6.20 cm on the 5th day, and almost covering the entire bottom of the plate on the 7th day. After 5 days of colony culture, a sparse, dark green conidia layer was produced on it, and it was distributed in a ring ( figure 1 A), the observation of the microscopic section shows that after cultivating for 5 days, the hyphae have branched, septum is arranged, smooth ( figure 1 B), where the bar value in A is 1.5cm, and the bar in B is 15μm.
[0034] Molecular level identification: Use the fungal genome extraction kit (Tiangen Biochemical Technology Co., Ltd.) to extract the genomic DNA of the pathogenic fungus according to the operating instructions. The general primers are as follows:
[0035] ITS1 (5'-GTTTCCGATAGGTGAACCTGC-3');
[0036]ITS4 (...
Embodiment 3
[0038] Example 3: Bioassays
[0039] The preserved strains were activated and cultured on a PDA slant at 25°C and photoperiod 12L:12D for 7 days until sporulation, and the conidia of the activated strains were transferred to 20 mL of PDA liquid medium (without adding agar). In a Erlenmeyer flask with PDA medium), shake (~150r / min) culture at 25°C for 2 days, then transfer 1 mL of bacterial liquid to 50 mL of culture liquid and continue culturing for about 1 day. Put the obtained mycelia liquid on the rice which has been sterilized by steam damp heat to an appropriate degree of maturity according to the ratio of 1 / 10 (v / w), and after fully mixing, spread the bacterium rice evenly in a sterilized petri dish with a diameter of 15 cm ( 100g rice / dish), at 25°C and 12L: 12D conditions, ferment and produce spores for 5-7 days, after the surface of the rice grains is covered with spores, dry them with air at 33°C for 2 days, and use the spore powder produced on the rice grains with ...
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