Application of Small Molecular Compounds in Inhibiting the Secretion System of Black Rot Fungus hrp III
A technology of small molecule compounds and black rot bacteria, applied in the field of agriculture, can solve the problems of unreported substances and achieve good results
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Embodiment 1
[0028] The detection of embodiment 1 compound to three types of fluorescence activity
[0029] After the tested strains Xcc 8004 / 0241lux and ΔhrpX / 0241lux were cultured overnight, measure the OD 600 Collect the bacterial liquid, then wash with XZM medium and resuspend the bacterial cells to OD 600 Adjust to 1. According to the starting cell OD 600 =0.15 concentration was transferred to XZM medium, and then the bacterial solution was dispensed and added to a 96-well plate, and 100 μL of bacterial solution was added to each well. Using DMSO as a control, each small molecule was added to a 96-well plate in a certain amount (xerophyllin G and carawayin B were both 5 μM, and the final concentration of xylose was 80 μM, and the three small molecules were combined according to their The respective amounts were added together). Each sample was replicated three times. After the addition, the 96-well plate was covered, and the edge of the plate was sealed with sealing glue, and plac...
Embodiment 2
[0031] The detection of embodiment 2 pathogenicity
[0032] The host plant used in the experiment was carrot seedlings all over the body (the species name is Raphanus sativus var. radiculac v. Manshenhong), and the inoculation method used was leaf-cutting method. The Xcc 8004 wild-type strain was cultured in NYGB medium at 28°C for 15-18h, and diluted to OD with NYGB supplemented with a certain concentration of compounds 600 =0.001, use sterilized scissors to soak in the bacterial solution for 5s, cut healthy leaves 1-2cm away from the leaf tip and cut to the central axis in the direction of the vertical veins, stay for 5s, and inoculate the plants at 25-30°C for 10d Observation results.
[0033] Pathogenicity tests showed that the pathogenicity of Xcc 8004 added with these three compounds was significantly reduced compared with that without addition (see figure 2 and image 3 ). The effect of adding a combination is better than three separate ones.
Embodiment 3
[0034] The measurement of growth curve in the leaf of embodiment 3
[0035] In this experiment, the plate number colony method was used for detection, and carrots with good growth and basically the same leaf size were selected for leaf-cut inoculation. Each sample was inoculated with at least 50 leaves to ensure the adoption of later experiments. After the inoculation, samples were taken on the 1st, 3rd, 5th, 7th, and 9th day respectively, and 9 leaves were picked randomly, and each 3 leaves was regarded as a group, and repeated at least 3 times.
[0036]For each sampling, 9 leaves of similar size were randomly selected from each sample, and then divided into 3 groups and marked, with 3 leaves in each group. The collected leaves are disinfected with medical disinfectant alcohol in the ultra-clean bench, mainly to wipe off the dust on the surface of the leaves (be careful when wiping, try to be as light as possible, to avoid artificial damage to the leaves). Dry the leaves, p...
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