Pseudomonas and application thereof
A pseudomonas and strain technology, applied in the field of microorganisms, can solve the problems affecting the growth and development of fruit trees, apple yield and quality, apple leaf shedding, environmental pollution, etc., and achieves good isolated leaf control effect, convenient application, and simple preparation process. Effect
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Embodiment 1
[0024] Isolation and identification of Pseudomonas sp. of the present invention
[0025] A. Isolation and preservation of Pseudomonas sp.
[0026] Step 1) Specimen collection: Collect the rhizosphere soil of healthy apple trees in Penglai District, Yantai City, Shandong Province, China, and put them in sterile bags for later use.
[0027] Step 2) Separation and purification of Pseudomonas sp.: Air-dry the collected soil samples and grind them. Weigh 10 g of the soil sample and put it into an Erlenmeyer flask, add 90 mL of sterile water, shake on a shaker for 30 minutes, and obtain a suspension of the soil sample. Draw 1mL of the suspension, add it into a small test tube filled with 9mL of sterile water, mix well, and obtain a concentration of 10 -2 The soil sample dilution solution, using the same method to obtain a concentration of 10 -5 、10 -6 soil sample dilution. Draw 100 μL of each of the two concentrations of dilutions and spread them on the NA medium with a coating...
Embodiment 2
[0039] Antibacterial activity analysis of Pseudomonas sp. of the present invention
[0040] The antibacterial activity analysis of the described Pseudomonas sp. was carried out by the plate confrontation growth method: with the apple leaf leaf spot and apple brown leaf spot as the indicator bacteria, the activated indicator bacteria cake was punched with a 6mm punch and placed In the center of the plate, inoculate Pseudomonas sp by spot-joining method, the distance between the two bacteria is about 4.5cm, and on the diameter of one line, the control group is only inoculated with the indicator bacteria cake, repeated 3 times, and cultured in the dark at 28°C. After the control group covered the plate, observe the growth of pathogenic bacteria in the test group, and measure the antibacterial zone ( image 3 , Figure 4 ,Table 2).
[0041] Table 2. Antibacterial activity analysis of Pseudomonas sp. against two indicator bacteria
[0042] Indicator bacteria Control ...
Embodiment 3
[0044] Antibacterial activity analysis of fermentation filtrate of Pseudomonas sp. of the present invention
[0045] After activation, the Pseudomonas sp. of the present invention was inserted into a 250mL conical flask containing 100mL fermentation medium at a volume ratio of 2%, and the fermentation broth was obtained by shaking at 30°C and 200rpm for 72h, and the fermentation broth was subjected to 5000r / min After centrifugation for 10 min, the supernatant was filtered and sterilized with a 0.22 μm microporous filter to obtain a fermentation filtrate. The fermentation filtrate was mixed with the sterilized PCDA medium at about 50°C in a volume ratio of 1:50, and then poured into the plate. Using the apple leaf leaf spot and apple brown leaf spot as the indicator bacteria, use a 6mm punch to punch out the activated ones. The indicator bacteria cake was placed in the center of the plate, and the growth rate method was used to analyze the antibacterial activity of the fermenta...
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