A kind of Pseudomonas fluorescens bacterial strain and its application
A technology of Pseudomonas fluorescens and strains, applied in the field of microorganisms, can solve the problems of waste of resources, pollution of the environment, no treatment methods, etc., achieve obvious social and economic benefits, and avoid environmental pollution
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Embodiment 1
[0020] Embodiment 1. The construction method and process of the Pseudomonas fluorescens strain zc-C3-V of the present invention are as follows:
[0021] 1. Take biocontrol Pseudomonas C3 and Pseudomonas fluorescens V as the starting bacteria, cultivate with modified and KB medium to logarithmic phase, remove the supernatant by centrifugation, and use 20mg / ml lysozyme solution for precipitation at 30°C After treatment for 1.5h, protoplasts were obtained.
[0022] 2. Dilute the protoplasts with SMM and pure water respectively, and then plate them to calculate the protoplast formation rate.
[0023] The protoplast formation rate of Pseudomonas fluorescens V is 75%, such as figure 1 As shown, the biocontrol Pseudomonas C3 protoplast formation rate was 73%, as shown in figure 2 As shown, the 8-h protoplast formation rate was 88%; it can be seen that the cell wall of C3 is not easily ruptured. figure 1 and figure 2 The picture on the left is diluted with water 10 6 Coating ...
Embodiment 2
[0027] Example 2. Identification of Pseudomonas fluorescens strain zc-C3-V
[0028] Amplify the 16S rRNA of the fusion with universal primers, and send it to Qingdao Qingke Zixi Biotechnology Co., Ltd. for sequencing. , its 16S rRNA is 99% similar to Pseudomonas fluorescens sp. f2 (2012), so it is determined that the constructed fusant is a Pseudomonas fluorescens strain.
Embodiment 3
[0029] Example 3 Experiment on acid production ability of Pseudomonas fluorescens strain zc-C3-V
[0030] The obtained strains zc-C3-V were all transferred to the LB plate with the starting bacteria Pseudomonas fluorescens V as a control, and different single clones were taken for seed culture, and the seed culture conditions were seed medium (g / L). ): glucose 20.0, corn steep liquor 10.0, urea 2.0, KH2PO4 2.0, MgSO4 7H2O 0.5 pH 7.0. Prepared according to the formula, and packed into a triangular bottle, 25ml / 250ml bottle. Then put it in a shaker room at 30°C for 16 to 20 hours with shaking at 200 rpm. After the cultivation, it was transferred to the fermentation medium. The fermentation formula is: glucose 18%, corn steep liquor 2%, calcium carbonate 4%, pH 6.7-7.0. Sterilize in a sterilizer at 118°C for 20 minutes. The fermentation conditions were 260 rpm and 30 °C for 72 h. The acid production conversion rate was calculated according to the optical rotation value, and ...
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