New applications of melatonin in inhibition of plant oomycetes diseases, and novel plant oomycetes bactericide
A bactericide and melatonin technology, applied in the field of melatonin, can solve the problem of less melatonin, and achieve the effects of reducing the amount of use, improving the efficacy of pesticides, and weakening the ability to infect
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Embodiment 1
[0025] Embodiment 1 prepares medium for experiment
[0026] 1. Rye solid medium (RSA): ① Soak 60g rye grains in 300ml distilled water, 4°C for 36 hours to avoid deterioration; ② Pour out the soaking water and store in the refrigerator, add 100ml of distilled water was mashed for 1-2min, and the suspension was placed in a water bath at 50°C for 3h; ④ Filter the suspension with 3-4 layers of gauze and save the filtrate; Add 20g of sucrose, then add water to 1000ml; ⑥Finally add 12g of agar, autoclave at 121°C for 20min.
[0027] 2. Potato Dextrose Agar Medium (PDA): Weigh 44.3g of commercial PDA medium powder, dissolve it in 1L of distilled water, and sterilize under high pressure at 121°C for 20min.
Embodiment 2
[0028] Embodiment 2 bacteria living experiment
[0029] 1. Treatment of potato leaves, potato cubes and tobacco leaves with melatonin alone
[0030] Potato leaves, potato cubes and tobacco leaves were treated with melatonin, and then the lesion area of the host plants was counted. Potato leaves and potato cubes with a volume of 2cm×3cm×3mm were soaked with different concentrations of melatonin (1mM, 3mM, 6mM, 8mM and 10mM) for 3-5 minutes, and then 7mm infestation bacteria were inoculated into potato leaves and On the potato cubes, place them at 18° C. under the conditions of a photoperiod of 12h:12h, for 5 days. Soak the tobacco leaves with different concentrations of melatonin (1mM, 3mM, 4mM and 6mM) for 3-5 minutes, then inoculate the tobacco leaves with a 7mm block of tobacco black shank bacteria, and place them at 27°C with a photoperiod of 12h:12h Next, 3 days.
[0031] According to the statistical lesion area (such as figure 1 Shown) can be found, with melatonin t...
Embodiment 3
[0036] Embodiment 3 germ in vitro experiment
[0037] 1. Treatment of potato late blight and tobacco black shank with melatonin
[0038] Configure rye solid medium, then add melatonin to the medium, and the final concentration of added melatonin is (1mM, 1.5mM, 2mM, 3mM and 5mM). Take 7mm potato infestans bacterial block and place it in the middle of the culture medium, seal it and place it in the dark at 18°C for 14 days; configure PDA medium, then add melatonin to the medium, the final concentration of melatonin added is (2mM, 4mM, 6mM, 8mM and 9mM). Take tobacco black shank bacterial block of 7 mm and place it in the middle of the medium, seal it and place it at 27°C in the dark for 18 days.
[0039] Observe the growth status of the two pathogens (such as Figure 5 , 6 Shown) It can be found that with the increase of melatonin concentration, the inhibition of bacterial growth is more and more obvious. Treat Potato infestans with melatonin, the drug concentration of I...
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