Application of a Bacillus fermented extract in the prevention and treatment of plant diseases and insect pests
A plant disease and extract technology, which is applied in the fields of application, plant growth regulators, and microbial-based methods, can solve the problems of pest resistance, harm to human health, and pesticide residues, and achieve simple production processes and low toxicity to humans and animals , The effect of low product cost
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Embodiment 1
[0012] Example 1: Bacillus horneckiae Preparation of DSM23495 fermentation extract
[0013] (1) Fermentation culture
[0014] According to the conventional culture method of microorganisms, select a small amount of strains stored on the slope of LB solid medium Bacillus horneckiae DSM23495, inoculated on the surface of the LB solid plate, cultured at 30°C for 2 days, as a strain for large-scale fermentation culture, ready for use.
[0015] LB solid medium: 1% tryptone, 0.5% yeast extract, 1% NaCl, 1% agar, distilled water, pH adjusted to 7.0.
[0016] Cut the above-mentioned bacteria on the surface of the LB solid plate, inoculate it into an Erlenmeyer flask of a sterile LB liquid medium, and culture it on a shaker at 30° C. and 120 rpm for 10 days for later use.
[0017] (2) Preparation of fermentation extract
[0018] The above LB liquid medium was extracted 3 times with ethyl acetate, the ethyl acetate extracts were combined and distilled under reduced pressure to obtain an extra...
Embodiment 2
[0019] Example 2: Antibacterial activity test
[0020] Using micro-dilution method to determine Bacillus horneckiae The antibacterial activity of the fermentation extract of DSM23495 against Botrytis cinerea, Fusarium wilt and apple rot disease.
[0021] 1) Preparation of bacterial suspension
[0022] After inoculating the test fungus on the surface of the PDA medium and culturing for 72 h at 28°C, draw 2 mL of sterile 0.85% NaCl solution (containing 0.25% Tween-20) to wash the culture, and gently scrape the colony with a glass spatula. Pipette an appropriate amount of bacterial suspension into a sterile test tube and adjust it to 0.5 McDonald's turbidity (equivalent to 1.5×10 8 CFU / mL) spare.
[0023] 2) Sample preparation
[0024] Take a certain amount of the sample to be tested (the elution component I prepared in Example 1), dissolve it in 100 μL of 50% DMSO, mix well, pipette 50 μL of the sample solution into another centrifuge tube, and then add 50 μL 50% DMSO to obtain a samp...
Embodiment 3
[0031] Example 3: Determination of insecticidal activity of Spodoptera exigua
[0032] Determined by leaf disc dipping method Bacillus horneckiae The insecticidal activity of the fermented extract of DSM23495 against the 3rd instar larvae of Spodoptera exigua.
[0033] Operation steps: (1) Cut the mesophyll of fresh cabbage leaves into small leaf discs of 1 cm × 1 cm, and prepare the sample (elution fraction II prepared in Example 1) with a 90% acetone solution to make a solution of a certain concentration; (2) Take 50 μL of sample solution and add it to a 24-well plate with 4 leaf discs, 8 replicates for each concentration, with 90% acetone solution as a blank control; (3) After the solvent evaporates, add Spodoptera exigua 3 Instar larvae, 1 per hole, covered with a lid, cultured in the dark at 35°C and RH40%; (4) After culturing for 48 hours, observe the feeding and death of the beet armyworm larvae, and calculate the corrected mortality and suppressed feeding rate . Correcte...
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