Application of AHL molecule to prevention and control of citrus bacterial canker as antibacterial synergist of chemical pesticide
A technology of citrus canker and chemical agents, which is applied in the field of xonopodispv.citri (Xa), can solve problems such as threats to human and animal health, environmental pollution, and soil compaction, and achieve the effects of promoting green agricultural development, environmental friendliness, and environmental protection
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Embodiment 1
[0025] Example 1 Preparation of AHL Small Molecules
[0026] The seed solution of Pseudomonas aeruginosa cultured overnight was inserted into YEB medium, 28 o C. After culturing at 200 rpm for 24 hours, the culture supernatant was collected by centrifugation. The obtained supernatant is used for extraction of small molecules. Among them, the formula of YEB medium is: 0.5 g / LMgSO 4 •7H 2 O, 5 g / L Sucrose, 5 g / L Yeast Extract, 10 g / L Trypton, 5 g / L NaCl, pH 7.0.
[0027] The present invention adopts the conventional compound separation and extraction method to separate and extract the active substances in the fermentation broth. After collecting the supernatant of the culture medium by centrifugation, add an equal volume of ethyl acetate to the supernatant, place it at 4°C for 2 hours, then shake it at 20°C for 1 hour, extract it with a separatory funnel, and collect the upper layer. Then, the collected upper layer was separated and evaporated with a rotary evaporator, and ...
Embodiment 2
[0028] Example 2 The bactericidal and synergistic effect of small-molecule AHL mixed with 1% shenzimycin on Xac of citrus canker
[0029] Colony counting statistical method (CFU method) was used to measure the bactericidal synergistic effect of different concentrations of small molecule AHL extracts on Shenzimycin: the small molecule AHL was prepared and diluted to a certain concentration with methanol as a solvent, and cultured overnight to OD 600 =1.0 of Xac Bacterial solution, add 1% Shenzimycin diluted 200 times 100mg / ml, set a treatment without small molecule AHL as a positive control, and then set a series of small molecules with concentration gradients (40μM, 80μM, 160μM, 320μM, 500μM) Treatment of Molecular AHL Mixed with Shenzimycin. All processed in 28 o C. After culturing at 200rpm for 24 hours, dilute 10 4 times, take 200 μL of the bacterial solution to smear the plate (the medium is LB), and place it in a constant temperature incubator at 28°C for cultivation...
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