Escherichia coli for preventing and treating bacterial wilt and its application
A technology for Escherichia coli and bacterial wilt disease, applied in the field of agricultural biology, can solve problems such as Escherichia coli research that has not yet been reported.
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Embodiment 1
[0020] Embodiment 1: Bacteria carry out separation and purification
[0021] 1. Medium
[0022] LB agar medium (LB solid medium): peptone 10g, yeast extract 5g, NaCl 10g, agar 15g, with H 2 Dilute to 1000 mL with O, pH 7.0-7.2, and sterilize at 121°C for 20 minutes.
[0023] 2. Experimental steps
[0024] (1) Sample collection: Select a representative location (root soil of healthy plants in a field with bacterial wilt disease, location in Guangdong Province), shovel off the topsoil, and use a soil picking tool to collect a soil layer at a depth of 5 to 10 centimeters below the surface 5 samples were taken from each site, packed into bags and recorded.
[0025] (2) Preparation of soil dilution: Weigh 10 g of soil, put it into a conical flask filled with 90 mL of sterile water and oscillate fully to prepare a soil dilution with a concentration of 1:10. After the soil particles have settled, absorb 1mL of the supernatant and transfer it into a test tube filled with 9mL of st...
Embodiment 2
[0028] Embodiment 2: the plate antagonism test of isolated bacteria to R. solanacearum
[0029] 1. Medium
[0030] TTC solid medium: ① 10g of peptone, 1g of casein acid hydrolyzate (Casein Acid Hydrolysate), 5g of glucose, 15g of agar, with H 2 O was adjusted to 1000mL, pH6.8-7.2; sterilized at 121°C for 20 minutes to obtain solution I; Filter and sterilize with a 0.22 μm filter membrane to obtain solution II; ③ When solution I is cooled to about 45°C, add 0.5mL solution II to every 100mL solution I, and pour the plate.
[0031] TTC liquid medium: same as TTC solid medium, without agar and TTC.
[0032] 2. Experimental steps
[0033] 2.1 Plate Antagonism Test - Primary Screening
[0034] 2.1.1 Strain activation
[0035] Ralstonia solanacearum and bacterium carry out strain activation: solanacearum GMI1000 (ATCC) and the 384 strains of bacteria isolated in Example 1 were transferred to TTC solid medium plate and LB agar medium plate respectively, and were cultured at 28° C...
Embodiment 3
[0052] Embodiment 3: the biological control effect test of bacterial strain GZ-34
[0053] In order to test the biocontrol effect of the strain GZ-34, the fermentation broth of the strain GZ-34 was tested on the biocontrol effect of bacterial wilt on potted plants.
[0054] 1. Experimental steps
[0055] 1.1 Buy eggplant seedlings
[0056] Buy eggplant seedlings grown in greenhouses.
[0057] 1.2 Transplanting eggplant seedlings
[0058] Fill the flowerpots (size 170mm*160mm) with matrix soil, compact them slightly, and then move one eggplant seedling into each pot, press the roots slightly with two fingers after watering, and cultivate them in a greenhouse. Cultivate in pots for about 4 weeks.
[0059] 1.3 Activation of R. solanacearum and antagonistic bacteria and preparation of fermentation broth
[0060] Ralstonia solanacearum GMI1000 and antagonistic bacteria GZ-34 were activated respectively, and then picked single colony into TTC liquid medium and LB liquid medium sh...
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