Application of Fomepizole in Killing Pine Xylophilus
A technology of pine xylophilus and fomepizole, which is applied in the direction of nematicides, applications, biocides, etc., can solve the problems of no control methods, etc., and achieve the effects of low cost, clear action sites, and good application prospects
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Embodiment 1
[0015] Embodiment 1, fomepizole is to the inhibitory effect of pine wood nematode alcohol dehydrogenase
[0016] In the experiment of the inhibitory effect of fomepizole on pine wood nematode alcohol dehydrogenase, in order to obtain the total protein of pine wood nematode, about 3000 sterile pine wood nematodes were washed three times with sterile distilled water, centrifuged at 3000rpm for 5 minutes, discarded Then, the collected pine wood nematodes were crushed, suspended in 100 μL of physiological saline, centrifuged at 12000 rpm at 4 °C for 30 minutes, and the supernatant was taken as the enzyme solution for testing. Alcohol dehydrogenase activity assay reaction system: 100mM Tris-HCl buffer (pH=8.3), 5mM NAD + , 50 μL of the enzyme solution to be tested, 100 mM ethanol, 0.02 mM fomepizole. Immediately after mixing, measure the UV absorption value at 340nm, and calculate the rate of change of the absorption value per unit time. The reaction system without fomepizole was...
Embodiment 2
[0018] Embodiment 2. Effect of Fomepizole on the movement and reproduction of pine xylophilus.
[0019] In the experiment of the effect of fomepizole on the movement and reproduction of pine xylophilus, about 3000 pine wood nematodes were soaked in 0.02mM fomepizole solution and incubated at 25°C for 24 hours. Under the same conditions, B. xylophilus soaked in sterile water was used as a control. Each experiment was repeated three times.
[0020] In the experiment of the effect of fomepizole on the movement of B. xylophilus, the number of head swings per minute was used to evaluate the strength of B. xylophilus. The results showed that the fomepizole-treated B. xylophilus swayed its head 16 times per minute, while the control B. xylophilus swayed 46 times. This indicated that the motility of B. xylophilus in the fomepizole-treated group was significantly weaker than that in the control group.
[0021] In the experiment of the effect of fomepizole on the reproduction of pine...
Embodiment 3
[0022] Embodiment 3, determination of the activity of fomepizole against pine wood nematodes.
[0023] In the bioassay experiment of fomepizole killing pine wood nematodes, 400 μL of different concentrations of fomepizole solutions (0.010mM-0.080mM) were used to resuspend pine wood nematodes in a sterile 24-well plate to adjust the number of pine wood nematodes in the test system. About 200 pine wood nematodes were treated with sterile distilled water as a negative control; after 24 hours of incubation at 25°C in the dark, observe the morphology of pine wood nematodes under a stereo microscope, calculate the number of pine wood nematodes that survived and died, and repeat the above experiment ; B. xylophilus under the field of vision is dead if it is stiff and motionless, otherwise it is survival. The 24-hour corrected mortality of pine xylophilus under different gradients is calculated respectively. The results are statistically analyzed with SPSS17.0. The calculation formula ...
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