Bacillus tequilensis and application thereof
A technology of bacillus and bacterial liquid, applied in the direction of application, bacteria, edible seed preservation, etc., can solve the problems of lack of efficient degradation or treatment methods, achieve good results, prevent grain mildew, and low cost
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Embodiment 1
[0031] Isolation and Purification of Bacillus Tequila D5-8
[0032] Blattella germanica is a strain raised in the Laboratory of Vector Organisms and Insect-borne Diseases of Shandong First Medical University. It has been continuously bred in the laboratory for 15 years. When isolating Bacillus Tequila, first rinse the body surface of the adult Blattella germanica with 75% alcohol for 3 times, 2 minutes each time, rinse with sterile water for 3 times and then dissect to obtain the complete digestive head of the German cockroach.
[0033] 1. Sterilization
[0034] All experimental equipment such as petri dishes, centrifuge tubes, test tubes, gun tips, and sterile water were sterilized by high pressure at 0.1MPa, 121°C, and sterilized for 30 minutes.
[0035] 2. Grinding
[0036] Cut the digestive tract of the German cockroach into pieces, put it into a sterile centrifuge tube, add 1ml of sterile water, and grind it thoroughly with a grinding rod.
[0037] 3. Serial dilution ...
Embodiment 2
[0051] Study on the Culture Conditions of Bacillus Tequila D5-8
[0052] 1. Temperature
[0053] Bacillus tequila D5-8 was inoculated in LB medium with pH 7.0, and cultured on a shaker with a rotation speed of 220r / min under the temperature conditions of 15°C, 25°C, 30°C, 35°C, and 40°C respectively. After 24 hours, samples were taken to measure the light absorption value (OD) at a wavelength of 600nm. 600 ). See Table 1, compare the OD of Bacillus tequila culture solution under various temperature conditions 600 It can be seen that the optimum culture temperature of Bacillus tequila D5-8 is 30°C, and the suitable growth range is between 25°C and 35°C.
[0054] The influence of table 1 temperature on the growth of Bacillus tequila
[0055] temperature OD 600
15℃ 1.056±0.055d 20℃ 1.227±0.099c 25℃ 1.397±0.058b 30℃ 1.597±0.054a 35℃ 1.514±0.119ab 40℃ 1.270±0.114c
[0056] 2.pH value
[0057] Bacillus tequila was respectively i...
Embodiment 3
[0068] Prepare Bacillus tequila D5-8 bacterium liquid, specifically comprise the following steps:
[0069] (1) Select the standard solid LB medium, adjust the pH value to 6.5, and inoculate the Bacillus tequila D5-8 strain after the plate culture dish is cooled to room temperature, and cultivate it for 1 day at a temperature of 25°C to obtain amplified Proliferated Bacillus Tequila D5-8 colony;
[0070] (2) Select the standard liquid LB medium, adjust the pH value to 6.5, and inoculate 1 cm per 1 L of standard liquid LB medium 2 Step (1) The Bacillus tequila D5-8 colonies propagated on the standard solid LB medium were cultivated for 1 day on a shaking table with a rotation speed of 220 rpm at a temperature of 25° C. to obtain a culture solution;
[0071] (3) Refrigerate the culture solution obtained in step (2) at 8°C for 8 hours, take it out, let it stand at room temperature, and then add water to dilute it. The volume ratio of culture solution to water is 1:100 to obtain T...
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