A kind of pantoea pineapple and its agent and application
A technology of Pantoea pineapple and microbial agent, which is applied in the field of plant protection, can solve the problem of less Pantoea pineapple, etc., and achieve the effects of lower production costs, easy production, and fast growth
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Embodiment 1
[0026] Example 1 Screening and identification of colloid anthracnose pathogen and Fusarium oxysporum antagonistic bacteria
[0027] 1. Screening of antagonistic bacteria
[0028] a. Isolation and purification of antagonistic bacteria
[0029] Take 10g of the rhizosphere soil sample of the diseased orchid plant and add it to a triangular flask filled with 90mL sterile water and 30-50 glass beads to make a suspension with a concentration of 1:10, and shake it in a shaker at 28°C and 180rpm After 20-30 minutes, place the bacterial solution in a water bath at 75-85°C for 12-18 minutes. Transfer 1 mL of the soil suspension into a test tube filled with 9 mL of sterile water to make 10 -2 The bacterial suspension was diluted successively to make 10 -3 、10 -4 、10 -5 、10 -6 bacterial suspension. After serial dilution, pipette 100 μL respectively for a dilution factor of 10 -3 -10 -6 The bacterial suspension was evenly spread on the separation medium-beef extract-peptone medium...
Embodiment 2
[0047] Embodiment 2 utilizes Pantoea pineapple P.ananatis H2 to prepare microbial bacterial agent
[0048] Pick a ring of H2 colonies on the LB plate or slant and transfer it to the test tube for activation, and culture it on a shaker at 30°C for 16 hours with a rotation speed of 150rpm. The LB liquid medium in the test tube is: peptone 10g / L, yeast extract 5g / L, chlorine Sodium chloride 5g / L, pH 7.0; the activated bacterial solution was inoculated into a 250mL Erlenmeyer flask containing 100mL LB liquid medium according to the inoculum amount of 1%-3%, at a speed of 150rpm, and cultured on a shaker at 35°C for 10h-24h , when the bacteria content is greater than 2×10 8 cfu / mL or more can obtain the fermentation broth, which can be stored as a microbial agent.
Embodiment 3
[0049] Example 3 Antagonistic effect of Pantoea pineapple P.ananatis H2 on colloid anthracnose pathogen
[0050] 1. The microbial bacterial agent prepared in Example 2 was centrifuged at room temperature at 10000r / min for 20min, and filtered with a microporous membrane of 0.22 μm to obtain the antagonistic bacteria filtrate;
[0051] 2. Take the antagonistic bacteria filtrate and sterilized PDA solid medium at about 50°C and mix them evenly at a ratio of 1:19 and pour them into a petri dish. After cooling, put a gelatinous anthracnose pathogenic bacteria cake with a diameter of 7mm in the center of the plate to Sterile water was used as the control, each treatment was repeated three times, and the diameter of the pathogenic bacteria colony was measured after 5 days.
[0052] Sterile filtrate antibacterial rate = (control colony diameter - treated colony diameter) / (control colony diameter - 0.7) × 100%.
[0053] The result is as Figure 5 It was shown that the antibacterial...
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