Method for culturing bacillus subtilis fungicide and composite fertilizer containing fungicide

A technology of Bacillus subtilis and bacteria agent, which is applied in the field of compound fertilizer and its preparation, can solve the problems of impact and low microbial degradation efficiency, and achieve the effects of improving disease resistance, saving money, and reducing dependence on chemical fertilizers

Inactive Publication Date: 2017-11-07

JIANGSU FOOD & PHARMA SCI COLLEGE

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AI-Extracted Technical Summary

Problems solved by technology

The Netherlands, Japan, Germany, and the United States have successively invested a large amount of funds in the research and application of soil pollution recovery, but the use of microorganis...

Abstract

The invention discloses a composite fertilizer containing a bacillus subtilis fungicide and a preparation method of the composite fertilizer and belongs to the technical field of fertilizers. The composite fertilizer is composed of the following raw materials in parts by weight: 50-80 parts of humus, 40-60 parts of sludge, 35-50 parts of chicken manure, 30-40 parts of straws, 20-30 parts of urea, 20-30 parts of calcium superphosphate, 20-30 parts of potassium chloride, 15-20 parts of water, 5-10 parts of bacillus subtilis fungicide, 5-10 parts of bacillus megatherium, 5-10 parts of rhodococcus rhodococcus powder and the balance of EM bacteria. The novel composite microbial fertilizer disclosed by the invention has the effects of improving physical and chemical properties of soil, improving the nutrient supplying capability of soil, reducing dependence of crops on the chemical fertilizer and inhibiting plant diseases and insect pests. Therefore, the novel composite microbial fertilizer and the preparation method thereof are suitable to be popularized and used in the field of fertilizers.

Application Domain

SuperphosphatesBacteria +6

Technology Topic

FungicideRhodococcus +14

Examples

Experimental program(3)

Example Embodiment

[0029] Example 1

[0030] A compound fertilizer containing Bacillus subtilis inoculants, consisting of the following raw materials by weight: 50 kg of humus, 40 kg of silt, 35 kg of chicken manure, 30 kg of straw, 20 kg of urea, 20 kg of superphosphate, 20 kg of potassium chloride, 15 kg of water, and Bacillus subtilis 5kg, Bacillus megaterium 5kg, Rhodococcus powder 5kg, Bacillus 1.2kg, Yeast 1.2kg, Lactobacillus 1.2kg.

[0031] In this embodiment, the preparation process of the Bacillus subtilis inoculum is as follows:

[0032] (1) Use a sterile pipette to draw 0.3-0.5 mL of sterile normal saline, drop it into the strain preservation tube, and shake it slightly until it is evenly suspended, add 0.2 mL of the bacterial solution to the beef extract peptone liquid medium, and shake at 35°C Cultivate for 1-2 days to get the bacteria liquid;

[0033] (2) Transfer the bacterial liquid obtained in step (1) to the beef broth peptone culture medium, and place the beef broth peptone culture medium in a thermostat at 35°C for 1 day to obtain Bacillus subtilis colonies;

[0034] (3) Inoculate a loop of Bacillus subtilis colonies obtained in step (2) on beef extract peptone culture medium for streaking and purification, and subject the purified Bacillus subtilis colonies to mutagenesis breeding to obtain Bacillus subtilis strains. The obtained Bacillus subtilis strain was inoculated on the beef extract peptone slant medium, and the beef extract peptone slant medium was cultured at 35° C. for 1 day for use;

[0035] (4) Insert the Bacillus subtilis strain on the beef extract peptone slant medium in step (3) into an Erlenmeyer flask containing a 30mL shake flask fermentation medium at an inoculum of 3% (v/v) for 6 hours. Obtain the fermentation broth of Bacillus subtilis;

[0036] (5) The Bacillus subtilis fermentation broth in step (4) is subjected to pilot-scale fermentation to obtain the Bacillus subtilis pilot-scale fermentation broth, and the obtained Bacillus subtilis pilot-scale fermentation broth is centrifuged, filtered and dried. The Bacillus subtilis agent is obtained.

[0037] In this embodiment, in the step (4), the Erlenmeyer flask containing the shake flask fermentation medium was cultured for 6 hours under the conditions of a rotation speed of 200 r/min and a pH of 6.

[0038] The invention also provides a method for preparing a compound fertilizer containing Bacillus subtilis inoculum, and the specific steps are as follows:

[0039] (1) Weigh the raw materials according to the weight ratio of the above raw materials, and set aside;

[0040] (2) Mix the humus, sludge, chicken manure and straw prepared in step (1) uniformly to obtain a mixture. Grind the obtained mixture with a grinder, and pass the ground mixture through a 100-mesh sieve to obtain a mixed powder ;

[0041] (3) Mix uniformly the urea, superphosphate and superphosphate prepared in step (1) to obtain a compound fertilizer, and mix the obtained compound fertilizer and the mixed powder in step (2) evenly to obtain a mixture Material, spare

[0042] (4) Add the prepared Bacillus subtilis inoculum, Bacillus megaterium and Rhodococcus powder in step (1) into a container filled with water and stir evenly to obtain a combined bacteria homogenate;

[0043] (5) Spray the homogenate of the combined bacteria prepared in step (4) evenly on the mixture to obtain semi-finished microbial fertilizer, and place the semi-finished microbial fertilizer at 32°C for drying treatment to obtain microbial fertilizer.

Example Embodiment

[0044] Example 2

[0045] The compound fertilizer containing Bacillus subtilis inoculum is composed of the following raw materials by weight: 60 kg of humus, 45 kg of silt, 40 kg of chicken manure, 35 kg of straw, 25 kg of urea, 25 kg of superphosphate, 25 kg of potassium chloride, 16 kg of water, and Bacillus subtilis 6kg, Bacillus megaterium 6kg, Rhodococcus powder 6kg, Bacillus 2.2kg, Yeast 2.2kg, Lactobacillus 2.2kg.

[0046] In this embodiment, the preparation process of the Bacillus subtilis inoculum is as follows:

[0047] (1) Use a sterile pipette to draw 0.3-0.5 mL of sterile normal saline, drop it into the strain preservation tube, and shake it slightly until it is evenly suspended, add 0.2 mL of the bacterial solution to the beef extract peptone liquid medium, and shake at 35°C Cultivate for 1-2 days to get the bacteria liquid;

[0048] (2) Transfer the bacterial liquid obtained in step (1) to beef broth peptone culture medium, and place the beef broth peptone culture medium in a thermostat at 35°C for 2 days to obtain Bacillus subtilis colonies;

[0049] (3) Inoculate a loop of Bacillus subtilis colonies obtained in step (2) on beef extract peptone culture medium for streaking and purification, and subject the purified Bacillus subtilis colonies to mutagenesis breeding to obtain Bacillus subtilis strains. The obtained Bacillus subtilis strain was inoculated on the beef extract peptone slant medium, and the beef extract peptone slant medium was cultured at 35° C. for 2 days for use;

[0050] (4) The Bacillus subtilis strain on the beef extract peptone slant medium in step (3) was fermented and cultured for 10 hours at an inoculum amount of 4% (v/v) into an Erlenmeyer flask containing a 40mL shake flask fermentation medium. Obtain the fermentation broth of Bacillus subtilis;

[0051] (5) The Bacillus subtilis fermentation broth in step (4) is subjected to pilot-scale fermentation to obtain the Bacillus subtilis pilot-scale fermentation broth, and the obtained Bacillus subtilis pilot-scale fermentation broth is centrifuged, filtered and dried. The Bacillus subtilis agent is obtained.

[0052] In this embodiment, in the step (4), the Erlenmeyer flask containing the shake flask fermentation medium is placed at a rotation speed of 300 r/min and a pH of 7 for 10 hours.

[0053] The invention also provides a method for preparing a compound fertilizer containing Bacillus subtilis inoculum, and the specific steps are as follows:

[0054] (1) Weigh the raw materials according to the weight ratio of the above raw materials, and set aside;

[0055] (2) Mix the humus, sludge, chicken manure and straw prepared in step (1) uniformly to obtain a mixture. Grind the obtained mixture with a grinder, and pass the ground mixture through a 150-mesh sieve to obtain a mixed powder ;

[0056] (3) Mix uniformly the urea, superphosphate and superphosphate prepared in step (1) to obtain a compound fertilizer, and mix the obtained compound fertilizer and the mixed powder in step (2) evenly to obtain a mixture Material, spare

[0057] (4) Add the prepared Bacillus subtilis inoculum, Bacillus megaterium and Rhodococcus powder in step (1) into a container filled with water and stir evenly to obtain a combined bacteria homogenate;

[0058] (5) Spray the homogenate of the combined bacteria prepared in step (4) evenly on the mixture to obtain a semi-finished microbial fertilizer, and place the obtained semi-finished microbial fertilizer at 33°C for drying treatment to obtain microbial fertilizer.

Example Embodiment

[0059] Example 3

[0060] The compound fertilizer containing Bacillus subtilis inoculants is composed of the following raw materials by weight: 70 kg of humus, 45 kg of silt, 40 kg of chicken manure, 35 kg of straw, 25 kg of urea, 25 kg of superphosphate, 25 kg of potassium chloride, 18 kg of water, and Bacillus subtilis 8kg, Bacillus megaterium 8kg, Rhodococcus powder 8kg, Bacillus 3kg, Yeast 3kg, Lactobacillus 3kg.

[0061] In this embodiment, the preparation process of the Bacillus subtilis inoculum is as follows:

[0062] (1) Use a sterile pipette to draw 0.3-0.5 mL of sterile normal saline, drop it into the strain preservation tube, and shake it slightly until it is evenly suspended, add 0.2 mL of the bacterial solution to the beef extract peptone liquid medium, and shake at 35°C Cultivate for 1-2 days to get the bacteria liquid;

[0063] (2) Transfer the bacterial liquid obtained in step (1) to beef broth peptone culture medium, and place the beef broth peptone culture medium in a thermostat at 35°C for 2 days to obtain Bacillus subtilis colonies;

[0064] (3) Inoculate a loop of Bacillus subtilis colonies obtained in step (2) on beef extract peptone culture medium for streaking and purification, and subject the purified Bacillus subtilis colonies to mutagenesis breeding to obtain Bacillus subtilis strains. The obtained Bacillus subtilis strain was inoculated on the beef extract peptone slant medium, and the beef extract peptone slant medium was cultured at 35° C. for 2 days for use;

[0065] (4) Put the Bacillus subtilis strain on the beef extract peptone slant medium in step (3) into an Erlenmeyer flask containing 350 mL shake flask fermentation medium at an inoculum of 5% (v/v) for 15 hours. Obtain the fermentation broth of Bacillus subtilis;

[0066] (5) The Bacillus subtilis fermentation broth in step (4) is subjected to pilot-scale fermentation to obtain the Bacillus subtilis pilot-scale fermentation broth, and the obtained Bacillus subtilis pilot-scale fermentation broth is centrifuged, filtered and dried. The Bacillus subtilis agent is obtained.

[0067] In this embodiment, in the step (4), the Erlenmeyer flask containing the shake flask fermentation medium is cultured for 15 hours under the conditions of a rotational speed of 300 r/min and a pH of 7.

[0068] The invention also provides a method for preparing a compound fertilizer containing Bacillus subtilis inoculum, and the specific steps are as follows:

[0069] (1) Weigh the raw materials according to the weight ratio of the above raw materials, and set aside;

[0070] (2) Mix the humus, sludge, chicken manure and straw prepared in step (1) uniformly to obtain a mixture. Grind the obtained mixture with a grinder, and pass the ground mixture through a 200-mesh sieve to obtain a mixed powder ;

[0071] (3) Mix uniformly the urea, superphosphate and superphosphate prepared in step (1) to obtain a compound fertilizer, and mix the obtained compound fertilizer and the mixed powder in step (2) evenly to obtain a mixture Material, spare

[0072] (4) Add the prepared Bacillus subtilis inoculum, Bacillus megaterium and Rhodococcus powder in step (1) into a container filled with water and stir evenly to obtain a combined bacteria homogenate;

[0073] (5) Spray the homogenate of the combined bacteria prepared in step (4) evenly on the mixture to obtain a semi-finished microbial fertilizer, and place the semi-finished microbial fertilizer at 34°C for drying treatment to obtain microbial fertilizer. fertilizer.

[0074] Compared with traditional microbial fertilizers, the new composite microbial fertilizer prepared in the present invention has obvious advantages: (1) The new composite stress-resistant microbial fertilizer in the present invention has a degrading efficiency of more than 70% for organophosphorus pesticides. The degradation rate of many different pesticide residues is also increased by more than 50%. (2) In the present invention, the cost of raw materials is controlled below 1,000 yuan/t, and the factory cost is controlled below 2,500 yuan/t, which greatly reduces the cost of the enterprise. (3) The microbial fertilizer in the present invention can not only improve the physical and chemical properties of the soil and increase the fertilizer supply capacity of the soil, but also reduce the dependence of crops on chemical fertilizers and inhibit the occurrence of diseases and insect pests. In summary, the novel composite microbial fertilizer prepared by the present invention is suitable for promotion and application in the fertilizer field.

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Description & Claims & Application Information

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Classification and recommendation of technical efficacy words

low cost
Improve physical and chemical properties

Method, apparatus and article for microfluidic control via electrowetting, for chemical, biochemical and biological assays and the like

InactiveUS7163612B2low cost

Owner:KECK GRADUATE INST OF APPLIED LIFE SCI

Method for culturing bacillus subtilis fungicide and composite fertilizer containing fungicide

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Examples

Example Embodiment

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

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