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Method for preparing Bacillus subtilis

A Bacillus subtilis, high temperature sterilization technology, applied in microorganism-based methods, biochemical equipment and methods, chemicals for biological control, etc., can solve the problems of high cost and large dosage, and achieve low production cost, Low energy consumption and no three wastes

Active Publication Date: 2010-02-24
海南利蒙特生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the wettable powder of Bacillus subtilis is commonly used in the market, which is usually prepared by conventional fermentation methods such as liquid deep fermentation, that is, only liquid medium is used to cultivate and reproduce microorganisms, and then filtered and dried to obtain Bacillus subtilis mother powder. Most of the spores of Bacillus subtilis are 20 billion live spores per gram, so it is used in a large amount to prevent crop diseases and insect pests, and the cost is high

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  • Method for preparing Bacillus subtilis

Examples

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Effect test

Embodiment 1

[0031] Strain cultivation: prepare potato sucrose liquid medium according to the recipe, adjust the pH value to 7.5, divide into six 500mL Erlenmeyer flasks, each bottle is 100mL, sterilize at 121°C for 30 minutes, and cool to room temperature. Take the strain of Bacillus subtilis, use an inoculation loop to pick a ring from the culture medium of the inclined test tube, inoculate it into a conical flask, put it on a shaker at 200 rpm, and cultivate it at 28°C for 40 hours. Vigorous growth, no bacteria form, the concentration is 1×10 9 live spores / ml for later use.

[0032] Seed tank cultivation: the volume of the seed tank is 200 liters, and the weight ratio of corn flour: soybean flour: water is 1:2:150 to prepare 120 liters of corn flour and soybean flour medium, adjust the pH value to 7.5, and steam at 120 to 130 ° C Bacteria for 30 minutes, cooled to room temperature, after the microscopic examination was normal, inoculate 1.2 liters of liquid bacteria in the flame protec...

Embodiment 2

[0036] Strain cultivation: Prepare beef juice peptone medium according to the formula, adjust the pH value to 7.0, divide into six 500mL Erlenmeyer flasks, each bottle is 100mL, sterilize at 125°C for 30 minutes, and cool to room temperature. Take the BS-0898 strain, use an inoculation loop to pick a ring from the strain in the culture medium of the inclined test tube, inoculate it into a conical flask, place it on a shaker at 200 rpm, cultivate it at 28°C for 20 hours, examine it under a microscope, and the strain Vigorous growth, no bacteria form, ready for use.

[0037] Seed tank cultivation: the volume is 200 liters, prepare 120 liters of corn flour and soybean flour culture medium according to the weight ratio of corn flour: soybean flour: water 1: 3: 200, adjust the pH value to 8.5, steam sterilize at 120-130° C. for 30 minutes, Cool to room temperature, after microscopic examination is normal, inoculate 1.2 liters of liquid bacteria in the flame protection, the inoculat...

Embodiment 3

[0041] Strain cultivation: prepare liquid beef juice peptone medium according to the formula, adjust the pH value to 6.5, divide into six 500mL Erlenmeyer flasks, each bottle is 100mL, sterilize at 121°C for 30 minutes, and cool to room temperature. Take the strain of Bacillus subtilis, use an inoculation loop to pick a ring from the strain of the inclined test tube culture medium, inoculate it into a conical flask, place it on a shaker at 200 rpm, and cultivate it at 30°C for 40 hours, and examine it under a microscope. Vigorous growth, no bacteria form, ready for use.

[0042] Seed tank cultivation: the volume of the seed tank is 200 liters, corn flour by weight: soybean flour: glucose: water is 1:2:1:200 to prepare 120 liters of corn flour soybean flour glucose medium, adjust the pH value to 7.8, 120~ Steam sterilize at 130°C for 30 minutes, cool to room temperature, and inoculate 1.2 liters of liquid strains under flame protection with an inoculation concentration of 0.6×10 ...

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Abstract

The invention discloses a method for preparing Bacillus subtilis, which adopts a liquid-solid two-phase combined fermentation process, i.e. seed solution is cultured in the liquid, is inoculated to asolid culture medium after reaching proper concentration without a filtering step and insusceptible to pollution, and the concentration of the prepared Bacillus subtilis is over 1*10<11> active spore / gram. The method has simple steps, little device investment, low production cost and no three wastes. The Bacillus subtilis prepared by the method has high concentration and good control effect on plant diseases and insect pests, can be used for controlling rice blast, vegetable gray mould, powdery mildew and oidium heveae steinm and has favorable application prospect.

Description

technical field [0001] The invention relates to a method for preparing Bacillus subtilis. Background technique [0002] Worldwide, more than 2 million tons of pesticides are produced in the world every year, mainly chemical pesticides, long-term use of chemical pesticides, pathogens and pests are prone to develop resistance, and some chemical pesticides have been proven to have carcinogenic, teratogenic, and mutagenic problems. At the same time, chemical pesticide residues also restrict the quality of agricultural products. [0003] In recent years, biopesticides have risen rapidly. Biological pesticides refer to pesticides that use bacteria, fungi, viruses, nematodes and their metabolites (agricultural antibiotics) as active ingredients. The technology, means, and level are relatively low, and the biopesticide itself has high requirements for technology, storage, and transportation, and the subsequent rise of pyrethroids has caused the development of biopesticides to ente...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01P3/00C12R1/125
Inventor 刘国忠张洁岩刘勇李新
Owner 海南利蒙特生物科技有限公司
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