Application of beta-nitro ethyl acrylate compound in preventing and/or treating plant diseases
A kind of ethyl nitroacrylate, plant disease technology, applied in the direction of application, plant growth regulator, botany equipment and methods, etc., to achieve strong inhibitory effect and reduce pathogenicity
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Embodiment 1
[0050] Example 1 compound is tested for hpa1 gene promoter activity
[0051] hpa1in PXO99 A The strain was activated on the plate of PSA medium for 3 days, and a single colony was picked and grown to OD in M210 liquid medium 600 About 0.6, 12000rmp / min centrifuge to collect the bacteria, resuspended in XOM 2 Medium and adjust OD 600 About 0.3, add the final concentration of 100μmol / L small molecule compound. Treat at 28°C and 200rmp / min for 15h, collect the bacteria by centrifugation and resuspend in 0.01mol / LPBS (pH=7.4) buffer, adjust OD 600 It is about 0.05, and the mean fluorescence intensity (MFI) of GFP is detected by flow cytometry. An equal volume of DMSO was used as a negative control; the experiment was repeated 3 times, and 3 parallel experiments were performed.
[0052] The experimental results show that the three compounds have a strong inhibitory effect on the activity of the hpa1 promoter, and the calculation method is DMSO%=MFI (化合物) / MFI (DMSO) , It can...
Embodiment 2
[0056] Embodiment 2 test compound is to wild-type bacterial strain PXO99 A growth effects
[0057] The test process is: under the condition of adding the compound to be tested, the PXO99 in the rich medium M210 and the T3SS induction medium XOM2 (XOM2 is a poor medium supplemented with 0.5% sucrose as a carbon source) were measured respectively. A growth curve. Xoo wild-type strain PXO99 A Streak on the PSA plate for activation, pick a single colony after 3 days and place it in M210 liquid medium, grow to OD 600 About 2.0, 12000rmp centrifuge for 2min to collect the bacteria, wash the bacteria once with sterile water and repeat the above operation, then resuspend in M210 or XOM2, adjust the OD 600 is 0.1. After adding 100 μmol / L of the compound to be tested, the bacterial solution was placed on a 96-well plate, and an equal volume of DMSO was used as a negative control, and 3 replicates were set for each treatment. Set the temperature at 28°C, and measure it with a fully ...
Embodiment 3
[0059] Example 3: Compounds 1-3, 1-7 and 1-8 on PXO99 A Impact of Hypersensitivity Response (HR) on Tobacco
[0060] Xoo wild-type strain PXO99 A Streak on the PSA plate for activation, pick a single colony and place it in M210 liquid medium after 3 days, and cultivate it to OD in the medium 600 About 2.0, centrifuge at 12000rmp for 2min to collect the bacteria, wash the bacteria once with sterile water, repeat the above operation, and adjust the OD 600 To 0.8, add 100 μmol / L of the three compounds to be tested and the positive control TS006 compound, and an equal volume of DMSO as the negative control, and treat at 28°C for 2 hours. Then use a needle-free syringe to inoculate on the tobacco that has been cultivated to the 4-leaf stage, continue to culture in the greenhouse after inoculation, and take pictures to observe the phenomenon after 24 hours.
[0061] Test results such as figure 2 shown. It can be seen from the figure that compounds I-3, I-7 and I-8 have obvious...
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