Bacillus licheniformis XNRB-3 and application thereof
A technology of Bacillus licheniformis and inoculum, which is applied in the field of agricultural microorganisms, can solve the problem of less Bacillus biological control agents, etc., and achieves the effects of reducing the obstacles of continuous cropping of apples, promoting growth and excellent effects.
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Embodiment 1
[0082] Example 1: Isolation and identification of bacterial strains
[0083] 1. Isolation and purification of strains:
[0084] The roots of healthy and vigorous apples were selected from replanted orchards in Jincheng City, Shanxi Province by the improved sticking root method, and the endogenous antagonistic bacteria were isolated and screened immediately. First rinse the roots with tap water for 5 minutes to remove the adhered soil particles, then immerse in 75% ethanol for 2 minutes for surface sterilization, then rinse with sterile water for 3 times, then soak in 5% sodium hypochlorite solution for 5 minutes, and finally rinse with sterile water Rinse 5 times. After proper drying, the surface-sterilized roots were cut into 1-cm root segments and transferred to LB medium to isolate endophytic bacteria. Four roots were placed in each Petri dish, and the Petri dish was kept at 37 °C for one week. In addition, apply the sterile water used to rinse the root system for the las...
Embodiment 2
[0108] Example 2: Optimization of fermentation process of strain XNRB-3 and preparation of fermentation-related products
[0109] 1. Optimization of liquid fermentation conditions:
[0110] The determination of the bacterial fermentation liquid adopts the turbidimetric method, that is, the fermentation liquid 10-15mL is centrifuged at 2000r / min for 2min, and the OD of the bacterial liquid is measured.600 value (the absorbance value of the solution at 600nm wavelength), combined with the results of single factor experiments and orthogonal experiments, Plackett Burman and Box Behnken optimized the composition of Bacillus licheniformis XNRB-3 liquid fermentation medium in response surface analysis (RSM) and fermentation conditions.
[0111] After optimization, the best liquid fermentation conditions of strain XNRB-3 are:
[0112] The composition of liquid fermentation medium: sucrose 21.03g / L, beef extract 8.54g / L, MgSO 4 1.0g / L, K 2 HPO 4 1.5g / L, KCl 1.0g / L; pH 7.5.
[011...
Embodiment 3
[0121] Example 3: Functional identification of strain XNRB-3 and fermentation-related products
[0122] 1. Broad-spectrum antibacterial effect of strain XNRB-3:
[0123] In order to further verify whether the XNRB-3 strain has a broad-spectrum antibacterial effect on plant-derived diseases, another four common pathogenic fungi were selected for further antibacterial tests. The result is as Figure 5 shown.
[0124] The results of confrontation test showed that XNRB-3 strain had a strong inhibitory effect on the growth of 4 kinds of pathogenic Fusarium mycelium. It also has a strong inhibitory effect on Phytophthora, Rhizoctonia solani, Alternaria and apple tree rot. Therefore, the strain XNRB-3 has a broad-spectrum antibacterial effect on plant pathogenic bacteria under the conditions of the in vitro petri dish test.
[0125] 2. Effects of fermentation-related products of strain XNRB-3 on the germination of Fusarium oxysporum spores:
[0126] The effect of the fermentatio...
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