Bacillus encapsulated granular fertilizer and its production process and application

Abstract

The application relates to the technical field of biological fertilizer, and particularly discloses a bacillus encapsulated granular fertilizer and a production process and application thereof. The bacillus encapsulated granular fertilizer comprises bacterial sludge and fertilizer granules; the bacterial sludge is wrapped on the outer surface of the fertilizer granules; the bacterial content of the bacterial sludge is 200-500 billion cfu / mL, and the viscosity is 8000-10000 mPa; the production process of the bacillus encapsulated granular fertilizer comprises the following steps: (1) seed liquid is first cultured, and then the seed liquid is subjected to fermentation culture to obtain fermentation liquid; (2) the fermentation liquid is placed in a centrifugal equipment to be centrifuged to obtain bacterial sludge; and (3) the bacterial sludge is wrapped on the outer surface of the fertilizer granules by using an encapsulation equipment to obtain the bacillus encapsulated granular fertilizer. The production process of the bacillus encapsulated granular fertilizer can reduce the loss of bacterial bodies and energy, and thus the bacillus encapsulated granular fertilizer with high encapsulation rate, low powder drop rate and excellent use effect is obtained.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological fertilizer, in particular to a bacillus encapsulated granular fertilizer and its production process and application. BACKGROUND

[0002] Microbial granular fertilizer is a kind of biological fertilizer containing microorganisms and trace elements, which is prepared by mixing or coating microorganisms into or on the surface of fertilizer granules through blending or coating method. In recent years, through continuous improvement of the production process of microbial granular fertilizer, microbial fertilizer has been given more comprehensive efficacy, and therefore has been unanimously recognized and widely favored by farmers.

[0003] There are mainly two kinds of preparation methods for traditional microbial granular fertilizer: one is to mix bacterial powder with fertilizer granules, and then use high-temperature drying and granulation process for preparation. The high-temperature drying process in this preparation method can cause denaturation or death of a large number of bacteria, resulting in a large amount of bacterial powder loss and difficulty in meeting product specifications. The other is to coat the bacterial powder prepared in advance on the outer surface of the fertilizer granules by using a drum granulation method. The bacterial powder in this preparation method is obtained by high-temperature spray drying of fermented bacterial liquid, which can cause a large number of bacterial death. In addition, the bacterial powder on the surface of the microbial granular fertilizer prepared by the above two methods is prone to fall off, and the microbial granular fertilizers are prone to stick together, resulting in difficulty in meeting the product specifications of the microbial granular fertilizer. Therefore, the preparation methods of traditional microbial granular fertilizer can cause waste of bacterial powder and energy loss, and the obtained microbial granular fertilizer has low viable bacterial content, which greatly increases the production cost of microbial granular fertilizer. SUMMARY

[0004] In order to reduce the waste of bacterial powder and energy loss and improve the viable bacterial content of microbial granular fertilizer, the present application provides a bacillus encapsulated granular fertilizer and its production process and application.

[0005] In the first aspect, the present application provides a bacillus encapsulated granular fertilizer, which adopts the following technical solution:

[0006] A bacillus encapsulated granular fertilizer, comprising bacterial sludge and fertilizer granules; the bacterial sludge is wrapped on the outer surface of the fertilizer granules; the bacterial content of the bacterial sludge is 200-500 billion cfu / mL, and the viscosity is 8000-10000 mPa.

[0007] The application obtains a bacillus encapsulated granular fertilizer with high viable cell content, good stability and high encapsulation efficiency by wrapping the bacterial slurry on the surface of the fertilizer particles and controlling the viable cell content and viscosity of the bacterial slurry in the above range. Compared with the related art, the bacillus encapsulated granular fertilizer does not have the phenomenon of powder falling, effectively avoids the waste of bacterial bodies, and contains a large amount of nutrients and metabolic products in the bacterial slurry, so that the prepared bacillus encapsulated granular fertilizer can maintain excellent viable cell content for a long time, thereby obtaining a long shelf life.

[0008] The application can promote the significant growth of the plant height and stem diameter of the tomato plant by applying the bacillus encapsulated granular fertilizer provided by the application to the soil and cultivating the tomato plant.

[0009] Preferably, the viable cell content of the bacterial slurry is 25-35 billion cfu / mL, and the viscosity is 9000-9500 mPa.

[0010] The application further controls the viable cell content and viscosity of the bacterial slurry in the above range, so that the bacillus encapsulated granular fertilizer has a longer shelf life and can make the plant height and stem diameter of the tomato plant grow faster.

[0011] In the application, the bacillus includes but is not limited to bacillus subtilis, bacillus velezensis and bacillus amyloliquefaciens.

[0012] Preferably, the bacillus is selected from bacillus subtilis, bacillus velezensis and bacillus amyloliquefaciens.

[0013] In a specific embodiment, the bacillus can be bacillus subtilis, bacillus velezensis, bacillus amyloliquefaciens or a mixture of bacillus subtilis, bacillus velezensis and bacillus amyloliquefaciens.

[0014] Preferably, the bacillus is a mixture of bacillus subtilis, bacillus velezensis and bacillus amyloliquefaciens.

[0015] In a second aspect, the application provides a production process of a bacillus encapsulated granular fertilizer, which adopts the following technical scheme:

[0016] The production process of the bacillus encapsulated granular fertilizer comprises the following steps:

[0017] (1) Seed liquid fermentation: first, culture the seed liquid, then ferment and culture the seed liquid to obtain a fermentation liquid;

[0018] (2) Centrifugation: centrifuge the fermentation liquid in a centrifugal device to obtain a bacterial slurry;

[0019] (3) coating: using a coating device to wrap the bacteria mud on the outer surface of the fertilizer particles to obtain the bacillus coating granular fertilizer.

[0020] The application adopts a seed liquid fermentation, centrifugation and coating production process to prepare a bacillus coating granular fertilizer. In the production process of the bacillus coating granular fertilizer, the fermentation liquid obtained by seed liquid fermentation is centrifuged to obtain a coating bacteria mud similar to a flow phase. The bacteria mud has a high bacteria content and appropriate viscosity. The bacteria mud is used for coating the fertilizer particles, which can not only retain the nutrients and metabolic products of the bacteria in the fermentation liquid, but also reduce the loss of microbial bacteria and energy, thereby reducing the waste of resources and energy. Compared with the related art, the production process of the bacillus coating granular fertilizer provided by the application effectively avoids the denaturation loss of bacteria in the high-temperature powder spraying process and the energy consumption in the spray drying process, thereby increasing the viable count content of the bacillus coating granular fertilizer, and a bacillus coating granular fertilizer with low production cost, high coating rate, low powder drop rate, long shelf life and good use effect is obtained.

[0021] Preferably, the rotating speed of the coating device is 10-15 r / min.

[0022] The rotating speed of the coating device is controlled in the range of 10-15 r / min, which can make the bacteria mud uniformly and firmly coated on the outer surface of the granular fertilizer, thereby effectively improving the coating effect of the bacillus coating granular fertilizer, reducing the powder drop rate of the bacillus coating granular fertilizer, and increasing the viable count of the bacillus coating granular fertilizer.

[0023] Preferably, in the centrifugation step, the centrifugation rate is 5000-6000 r / min, and the centrifugation time is 250-300 s.

[0024] By adjusting the centrifugation rate, centrifugation time and other parameters in the centrifugation step, the viable count of the bacteria mud can be increased, and the flowability and viscosity of the bacteria mud can be improved. It is found that by further controlling the centrifugation rate and centrifugation time in the above range, a bacteria mud similar to a flow phase with high viable count and good viscosity can be obtained. The bacteria mud similar to a flow phase is used for coating the granular fertilizer, and the obtained bacillus coating granular fertilizer has the advantages of high coating rate, low powder drop rate and good coating effect.

[0025] In some embodiments, the centrifugation rate can be 5000-5500 r / min or 5500-6000 r / min.

[0026] In a specific embodiment, the centrifugation rate can be 5000 r / min, 5500 r / min or 6000 r / min.

[0027] In a specific embodiment, the centrifugation time can be 250 s or 300 s.

[0028] Preferably, the displacement in the centrifugation step is 0.3-0.4 L.

[0029] Preferably, the fermentation culture employs LB medium and / or total water-soluble medium.

[0030] Further, the fermentation culture includes primary fermentation, secondary fermentation and tertiary fermentation.

[0031] Preferably, the culture seed liquid and the primary fermentation employ LB medium.

[0032] The formula of the LB medium is: yeast extract 5 g / L, tryptone 10 g / L, sodium chloride 10 g / L.

[0033] Preferably, the secondary fermentation and the tertiary fermentation employ total water-soluble medium.

[0034] The components of the total water-soluble medium are selected from the group consisting of glucose, sucrose, soluble starch, yeast powder, tryptone, soybean meal, soybean peptone, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, manganese sulfate, calcium chloride, ammonium sulfate and antifoaming agent.

[0035] Further, the components of the total water-soluble medium include the following components:

[0036] one or both of glucose and corn flour, preferably in an amount of 0.5-2 g / L, and

[0037] one or more of yeast powder, tryptone, ammonium sulfate, soybean meal, soybean peptone, preferably in an amount of 0.5-2 g / L, and

[0038] dipotassium hydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, manganese sulfate, calcium carbonate and antifoaming agent.

[0039] In some embodiments, the following components are used in an amount of: dipotassium hydrogen phosphate 0.2-1 g / L, potassium dihydrogen phosphate 0.2-1 g / L, magnesium sulfate 0.1-0.5 g / L, manganese sulfate 0.1-0.5 g / L, calcium carbonate 0.1-0.5 g / L, and antifoaming agent 0.2-0.5 g / L, according to the volume of the total water-soluble medium.

[0040] In a specific embodiment, the formula of the total water-soluble medium can also be: glucose 12 g / L, yeast powder 4 g / L, ammonium sulfate 5.15 g / L, dipotassium hydrogen phosphate 1 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.5 g / L, manganese sulfate 0.5 g / L, calcium carbonate 2 g / L, and antifoaming agent 0.3 g / L.

[0041] In a specific embodiment, the formula of the full water-soluble culture medium can be: glucose 12 g / L, yeast powder 4 g / L, ammonium sulfate 8.87 g / L, dipotassium hydrogen phosphate 1 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.5 g / L, manganese sulfate 0.5 g / L, calcium carbonate 2 g / L, antifoam agent 0.5 g / L.

[0042] In a specific embodiment, the formula of the full water-soluble culture medium can also be: corn flour 12 g / L, soybean peptone 8 g / L, soybean cake powder 8 g / L, ammonium sulfate 8.87 g / L, dipotassium hydrogen phosphate 1 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.5 g / L, manganese sulfate 0.5 g / L, calcium carbonate 2 g / L, antifoam agent 0.4 g / L.

[0043] Further, the fermentation step specifically comprises: inoculating the seed liquid into LB culture medium, and then inoculating the LB culture medium into a shaker for primary fermentation; after the primary fermentation is completed, inoculating the fermentation liquid into a full water-soluble culture medium for secondary fermentation and tertiary fermentation.

[0044] Preferably, the inoculation amount of the fermentation culture is 5-10%, the rotation speed is 150-180 r / min, and the temperature is 32-37℃.

[0045] Still further, the time of the primary fermentation is 6-8 h, the time of the secondary fermentation is 6-8 h, and the time of the tertiary fermentation is 19-28 h.

[0046] In the present application, the inoculation amount, rotation speed, temperature and fermentation time of the fermentation culture are controlled within the above ranges, which can realize the rapid growth and reproduction of Bacillus, thereby increasing the viable cell count in the fermentation product and increasing the viable cell count content of the bacterial slurry, so that the bacterial slurry is more suitable for preparing Bacillus coated granular fertilizer with high bacterial content.

[0047] Preferably, in the coating step, the weight ratio of the bacterial slurry to the fertilizer granules is (4-6):100.

[0048] In a third aspect, the present application provides the use of the Bacillus coated granular fertilizer in soil.

[0049] The Bacillus coated granular fertilizer provided by the present application can be applied to soil to cultivate plants. Through cultivation tests on tomatoes, it is found that the Bacillus coated granular fertilizer can significantly increase the height and diameter of tomatoes, thereby achieving the growth-promoting effect on tomato plants. Therefore, it is proved that the Bacillus coated granular fertilizer provided by the present application can adjust the microbial environment in the soil, realize the prevention and control of plant diseases and insect pests, and thereby ensure the rapid and normal growth of crops.

[0050] Preferably, the weight ratio of the bacillus encapsulated granular fertilizer to the soil is (0.8-1.2):2000.

[0051] In the present application, the bacillus is wrapped on the outer surface of the fertilizer granule, so that the bacillus microorganism can utilize the nitrogen in the air to convert the fertilizer granule, while reducing the volatilization and loss of the fertilizer granule, promoting the absorption and conversion of the plant to the fertilizer granule; in addition, the bacillus can also decompose the difficult soluble phosphorus and potassium components in the soil, and improve the soil hardening problem caused by chemical fertilizer. Therefore, the weight ratio of the bacillus encapsulated granular fertilizer to the soil in the present application is controlled within the above range, and the soil obtained by cultivating the soil is loose, and can hold a large amount of nutrients for a long time, so it has a good growth effect on tomato plants.

[0052] In a specific embodiment, the weight ratio of the bacillus encapsulated granular fertilizer to the soil is 1:2000.

[0053] In summary, the present application has the following beneficial effects:

[0054] 1. The present application provides a bacillus encapsulated granular fertilizer prepared from bacterial sludge and fertilizer granules, wherein the bacterial sludge is wrapped on the outer surface of the fertilizer granule, the bacterial content of the bacterial sludge is 20-50 billion cfu / mL, and the viscosity is 8000-10000 mPa. Compared with the biological granular fertilizer in the related art, the above-mentioned bacillus encapsulated granular fertilizer has the advantages of high viable bacterial content, no powder dropping phenomenon, good stability, and high encapsulation efficiency.

[0055] 2. The present application provides a production process of bacillus encapsulated granular fertilizer, comprising seed liquid fermentation, centrifugation, encapsulation and the like. The production process can obtain a flow phase-like encapsulatable bacterial sludge through centrifugation, which can be directly used for encapsulation of granular fertilizer. Therefore, the production process effectively avoids the denaturation loss of bacterial bodies in the high-temperature powder spraying process and the energy consumption in the spray drying process, reduces the waste of resources and energy, increases the viable bacterial content in the bacillus encapsulated granular fertilizer, and makes the use effect of the bacillus encapsulated granular fertilizer better.

[0056] 3. The present application uses an encapsulation device to prepare bacillus encapsulated granular fertilizer, and controls the rotation speed of the encapsulation device within the range of 10-15 r / min, so that the bacterial sludge can be uniformly and firmly wrapped on the outer surface of the granular fertilizer, effectively improving the encapsulation effect of the bacillus encapsulated granular fertilizer, reducing the powder dropping rate of the bacillus encapsulated granular fertilizer, and ensuring that the bacillus encapsulated granular fertilizer has a high viable bacterial count.

[0057] 4. By controlling the centrifugal speed and centrifugal time of the centrifugation step, the present application can adjust the viable count, fluidity and viscosity of the slurry. By controlling the centrifugal speed between 5000-6000 r / min and the centrifugal time between 250-300 s, a slurry close to a mobile phase can be obtained. Using the above-mentioned slurry close to a mobile phase to coat the fertilizer particles, a bacillus encapsulated granular fertilizer with high coating rate, low powder loss rate and good coating effect can be obtained.

[0058] 5. The bacillus encapsulated granular fertilizer prepared by the present application can be applied to the soil to regulate the bacterial community environment in the soil, achieve crop disease and pest control, and thus ensure the rapid and normal growth of crops. Through the exploration of the growth-promoting effect of bacillus encapsulated granular fertilizer on tomato plants, it is found that the growth-promoting effect of composite bacillus encapsulated granular fertilizer > bacillus velezensis encapsulated granular fertilizer > bacillus amyloliquefaciens encapsulated granular fertilizer > bacillus subtilis encapsulated granular fertilizer. DETAILED DESCRIPTION

[0059] The present application provides a bacillus encapsulated granular fertilizer. The production process of the bacillus encapsulated granular fertilizer includes the following steps:

[0060] (1) Seed liquid fermentation: the bacillus is placed in LB culture medium for 24 h culture, and the seed liquid is obtained after the culture is completed; then the seed liquid is inoculated into a 100 L fermenter at an inoculation amount of 5-10% for primary fermentation, the culture medium for primary fermentation is LB culture medium, the culture time for primary fermentation is 6-8 h, when the bacterial count density under microscopic examination reaches more than 100, the dissolved oxygen in the culture medium decreases to less than 30%, the seed liquid is transferred to a 1 ton fermenter for secondary fermentation at an inoculation amount of 5-10%, the culture medium for secondary fermentation is a full water-soluble culture medium, the culture time for secondary fermentation is 6-8 h, when the bacterial count density under microscopic examination reaches more than 100, the dissolved oxygen in the culture medium decreases to less than 30%, the seed liquid is transferred to a 10 ton fermenter for tertiary fermentation at an inoculation amount of 5-10%, the culture medium for tertiary fermentation is a full water-soluble culture medium, the culture time for tertiary fermentation is 19-28 h, when the spore production rate reaches more than 99%, the culture is completed, and the bacillus subtilis fermentation liquid is obtained. The rotation speed of seed liquid culture, primary fermentation, secondary fermentation and tertiary fermentation is 150-180 r / min, and the temperature is 32-37℃.

[0061] (2) Centrifugation: the fermentation liquid is placed in a butterfly type centrifugal equipment for centrifugation, the centrifugal speed is set to 5000-6000 r / min, the centrifugal time is 250-300 s, and the displacement is 0.3-0.4 L, and the slurry is obtained after centrifugation;

[0062] (3) Coating: the bacteria slurry is added into a storage tank, and then the bacteria slurry is sprayed into a cylinder containing fertilizer particles by using a water pump nozzle, and coating is performed at a cylinder rotation speed of 10-15 r / min, and the bacillus subtilis coated particle fertilizer is obtained after the cylinder coating.

[0063] The bacillus subtilis coated particle fertilizer produced by using the production process of the bacillus subtilis coated particle fertilizer is applied in soil; further, the weight ratio of the bacillus subtilis coated particle fertilizer to soil is (0.8-1.2):2000.

[0064] In the present application, the bacillus subtilis is bacillus subtilis, bacillus velezensis, bacillus amyloliquefaciens, or a mixture of bacillus subtilis, bacillus velezensis and bacillus amyloliquefaciens.

[0065] In the present application, the formula of the LB medium and the fully water-soluble medium are as follows, respectively:

[0066] The formula of the LB medium is: yeast extract 5 g / L, tryptone 10 g / L, sodium chloride 10 g / L, water 1 L.

[0067] The formula of the fully water-soluble medium is: one or both of glucose and corn flour in an amount of 0.5-2 g / L; one or more of yeast powder, tryptone, ammonium sulfate, soybean meal, and soybean protein peptone in an amount of 0.5-2 g / L; dipotassium hydrogen phosphate 0.2-1 g / L, potassium dihydrogen phosphate 0.2-1 g / L, magnesium sulfate 0.1-0.5 g / L, manganese sulfate 0.1-0.5 g / L, calcium carbonate 0.1-0.5 g / L, and defoaming agent 0.2-0.5 g / L.

[0068] In the present application, the preservation number of bacillus velezensis is CGMCC No. 20317, which has been disclosed in the ZL202110072888.X patent; bacillus subtilis and bacillus amyloliquefaciens are purchased from Beijing Century Arms Biological Engineering Co., Ltd.; the model of the defoaming agent is DDYD-625, which is purchased from Nanjing Daoda Chemical Co., Ltd.; the raw materials, reagents, solvents, etc. in the present application can be obtained by commercial purchase.

[0069] The present application is further described in detail below in combination with preparation examples, examples, comparative examples and detection tests.

[0070] Preparation Example

[0071] Preparation Example 1 provides a bacillus subtilis fermentation broth.

[0072] The preparation method of the above-mentioned Bacillus subtilis fermentation broth is as follows: the Bacillus subtilis is placed in an LB culture medium for culture, and the culture conditions are as follows: the temperature is 35°C, the rotation speed is 180 r / min, and the time is 24 h, and a seed liquid is obtained after the culture is completed; then the seed liquid is inoculated into a 100 L culture tank at a 10% inoculation amount for primary fermentation, the culture medium for the primary fermentation is an LB culture medium, and the culture conditions for the primary fermentation are as follows: the temperature is 35°C, the rotation speed is 150 r / min, and the time is 6-8 h, when the density of the bacterial number under microscopic examination reaches more than 100, and the dissolved oxygen in the medium reduces to less than 30%, the Bacillus subtilis is transferred into a 1 ton fermentation tank at a 10% inoculation amount for secondary fermentation, the culture medium for the secondary fermentation is a full water-soluble culture medium, and the culture conditions for the secondary fermentation are as follows: the temperature is 35°C, the rotation speed is 160 r / min, and the time is 6-8 h, when the density of the bacterial number under microscopic examination reaches more than 100, and the dissolved oxygen in the medium reduces to less than 30%, the Bacillus subtilis is transferred into a 10 ton fermentation tank at a 10% inoculation amount for tertiary fermentation, the culture medium for the tertiary fermentation is a full water-soluble culture medium, and the culture conditions for the tertiary fermentation are as follows: the temperature is 35°C, the rotation speed is 180 r / min, and the time is 19-28 h, when the spore production rate reaches more than 99%, the culture is ended, and the Bacillus subtilis fermentation broth is obtained.

[0073] In the preparation method of the above-mentioned Bacillus subtilis fermentation broth, the formula of the full water-soluble culture medium used is as follows: glucose 12 g / L, yeast powder 4 g / L, ammonium sulfate 5.15 g / L, dipotassium hydrogen phosphate 1 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.5 g / L, manganese sulfate 0.5 g / L, calcium carbonate 2 g / L, and defoaming agent 0.3 g / L.

[0074] Preparation Example 2

[0075] Preparation Example 2 provides a Bacillus velezensis fermentation broth.

[0076] The preparation method of the Bacillus velezensis fermentation broth is the same as that of the Bacillus subtilis fermentation broth, except that the Bacillus subtilis in the preparation method of the Bacillus subtilis fermentation broth is replaced by Bacillus velezensis, and thus the Bacillus velezensis fermentation broth is obtained.

[0077] In the preparation method of the above-mentioned Bacillus velezensis fermentation broth, the formula of the full water-soluble culture medium used is as follows:

[0078] Glucose 12 g / L, yeast powder 4 g / L, ammonium sulfate 8.87 g / L, dipotassium hydrogen phosphate 1 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.5 g / L, manganese sulfate 0.5 g / L, calcium carbonate 2 g / L, and defoaming agent 0.5 g / L.

[0079] Preparation Example 3

[0080] Preparation Example 3 provides a Bacillus amyloliquefaciens fermentation broth.

[0081] The preparation method of the Bacillus velezensis fermentation broth is the same as that of the Bacillus subtilis fermentation broth, except that Bacillus subtilis in the preparation method of the Bacillus subtilis fermentation broth is replaced by Bacillus amyloliquefaciens, so that the Bacillus amyloliquefaciens fermentation broth can be obtained.

[0082] In the preparation method of the Bacillus amyloliquefaciens fermentation broth, the formula of the total water-soluble culture medium used is: corn flour 12 g / L, soybean peptone 8 g / L, soybean cake powder 8 g / L, ammonium sulfate 8.87 g / L, dipotassium hydrogen phosphate 1 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.5 g / L, manganese sulfate 0.5 g / L, calcium carbonate 2 g / L, and defoaming agent 0.4 g / L.

[0083] Viable cell count detection

[0084] The effective viable cell counts in the Bacillus subtilis fermentation broth, the Bacillus velezensis fermentation broth and the Bacillus amyloliquefaciens fermentation broth obtained in Preparation Examples 1-3 are detected, and the detection results are shown in Table 1. The detection method of the effective viable cell count refers to GB20287-2006.

[0085] Table 1 Effective viable cell counts of the Bacillus fermentation broths obtained in Preparation Examples 1-3

[0086]

[0087]

[0088] Preparation Example 4

[0089] Preparation Example 4 provides a Bacillus subtilis slurry.

[0090] The preparation method of the Bacillus subtilis slurry is as follows: the Bacillus subtilis fermentation broth provided in Preparation Example 1 is placed in a butterfly centrifuge for separation, the centrifugal speed is set to 5000 r / min, the centrifugal time is 250 s, and the displacement is 0.3 L, so that the Bacillus subtilis slurry is obtained.

[0091] Preparation Example 5

[0092] Preparation Example 5 provides a Bacillus velezensis slurry.

[0093] The preparation method of the Bacillus velezensis slurry is as follows: the Bacillus velezensis fermentation broth provided in Preparation Example 2 is placed in a butterfly centrifuge for separation, the centrifugal speed is set to 5000 r / min, the centrifugal time is 250 s, and the displacement is 0.3 L, so that the Bacillus velezensis slurry is obtained.

[0094] Preparation Example 6

[0095] Preparation Example 6 provides a B. amyloliquefaciens slurry.

[0096] The preparation method of the B. amyloliquefaciens slurry is as follows: the B. amyloliquefaciens fermentation broth provided in Preparation Example 3 is subjected to separation in a butterfly centrifuge, the centrifugal speed is set to 5000 r / min, the centrifugal time is 250 s, and the displacement is 0.3 L, to obtain the B. amyloliquefaciens slurry.

[0097] Preparation Example 7

[0098] Preparation Example 7 provides a B. velezensis slurry.

[0099] The above preparation example is different from Preparation Example 5 in that the centrifugal speed in the preparation method of the B. velezensis slurry; the centrifugal speed in Preparation Example 7 is 5500 r / min.

[0100] Preparation Example 8

[0101] Preparation Example 8 provides a B. velezensis slurry.

[0102] The above preparation example is different from Preparation Example 5 in that the centrifugal speed in the preparation method of the B. velezensis slurry; the centrifugal speed in Preparation Example 8 is 6000 r / min.

[0103] Preparation Example 9

[0104] Preparation Example 9 provides a B. velezensis slurry.

[0105] The above preparation example is different from Preparation Example 5 in that the centrifugal time in the preparation method of the B. velezensis slurry; the centrifugal time in Preparation Example 9 is 300 s.

[0106] Preparation Example 10

[0107] Preparation Example 10 provides a B. velezensis slurry.

[0108] The above preparation example is different from Preparation Example 5 in that the displacement in the preparation method of the B. velezensis slurry; the displacement in Preparation Example 10 is 0.4 L.

[0109] Comparative Example

[0110] Comparative Example 1

[0111] Comparative Example 1 provides a B. velezensis slurry.

[0112] The above Comparative Example 1 is different from Preparation Example 5 in that the centrifugal speed in the preparation method of the B. velezensis slurry; the centrifugal speed in Comparative Example 1 is 4000 r / min.

[0113] Comparative Example 2

[0114] Comparative Example 2 provides a Bacillus velezensis slurry.

[0115] The above Comparative Example 2 differs from Preparation Example 5 in that the centrifugal speed in the preparation method of the Bacillus velezensis slurry; the centrifugal speed in Comparative Example 2 is 7000 r / min.

[0116] Comparative Example 3

[0117] Comparative Example 3 provides a Bacillus velezensis slurry.

[0118] The above Comparative Example 3 differs from Preparation Example 5 in that the centrifugal speed in the preparation method of the Bacillus velezensis slurry; the centrifugal time in Comparative Example 3 is 200 s.

[0119] Comparative Example 4

[0120] Comparative Example 4 provides a Bacillus velezensis slurry.

[0121] The above Comparative Example 4 differs from Preparation Example 5 in that the centrifugal speed in the preparation method of the Bacillus velezensis slurry; the centrifugal time in Comparative Example 4 is 400 s.

[0122] Viable cell count and water content detection

[0123] The effective viable cell count in the Bacillus slurry obtained in Preparation Examples 4-10 and Comparative Examples 1-4 was detected, and the detection results are shown in Table 2.

[0124] Table 2 Effective viable cell count of Bacillus slurry obtained in Preparation Examples 4-10 and Comparative Examples 1-4

[0125]

[0126]

[0127] According to the detection results in Table 2, the effective viable cell count in the Bacillus velezensis slurry obtained in Preparation Examples 4-10 is greater than 200 billion cfu / mL, and the viscosity is between 8000-10000 mPa, while the effective viable cell count in the Bacillus velezensis slurry obtained in Comparative Example 1 and Comparative Example 3 is less than 200 billion cfu / mL, and the viscosity of the Bacillus velezensis slurry obtained in Comparative Example 2 and Comparative Example 4 is greater than 10000 mPa·s. Therefore, by controlling the centrifugal speed to be between 5000-6000 r / min and the centrifugal time to be between 250-300 s, a Bacillus velezensis slurry with high viable cell count and moderate viscosity can be obtained, and a Bacillus velezensis coated granule with high coating rate, high viable cell count and low dust loss rate can be obtained by using the Bacillus velezensis slurry to coat the granular fertilizer.

[0128] Example

[0129] Example 1

[0130] Example 1 provides a Bacillus subtilis coated granular fertilizer.

[0131] The preparation method of the above-mentioned Bacillus subtilis coated granular fertilizer is as follows: 5 kg of Bacillus subtilis slurry (containing 200 billion cfu / mL) provided by Preparation Example 4 is loaded into a storage tank, the slurry is sprayed into a drum containing 100 kg of granular fertilizer by using a water pump nozzle, the rotation speed of the drum is 10 r / min, and the Bacillus subtilis coated granular fertilizer is obtained after drum coating.

[0132] Example 2

[0133] Example 2 provides a Bacillus velezensis coated granular fertilizer.

[0134] The preparation method of the above-mentioned Bacillus velezensis coated granular fertilizer is as follows: 5 kg of Bacillus velezensis slurry (containing 200 billion cfu / mL) provided by Preparation Example 5 is loaded into a storage tank, the slurry is sprayed into a drum containing 100 kg of granular fertilizer by using a water pump nozzle, the rotation speed of the drum is 10 r / min, and the Bacillus velezensis coated granular fertilizer is obtained after drum coating.

[0135] Example 3

[0136] Example 3 provides a Bacillus amyloliquefaciens coated granular fertilizer.

[0137] The preparation method of the above-mentioned Bacillus amyloliquefaciens coated granular fertilizer is as follows: 5 kg of Bacillus amyloliquefaciens slurry (containing 200 billion cfu / mL) provided by Preparation Example 6 is loaded into a storage tank, the slurry is sprayed into a drum containing 100 kg of granular fertilizer by using a water pump nozzle, the rotation speed of the drum is 10 r / min, and the Bacillus amyloliquefaciens coated granular fertilizer is obtained after drum coating.

[0138] Example 4

[0139] Example 4 provides a Bacillus velezensis coated granular fertilizer.

[0140] The preparation method of the above-mentioned Bacillus velezensis coated granular fertilizer is as follows: 5 kg of Bacillus velezensis slurry (containing 200 billion cfu / mL) provided by Preparation Example 5 is loaded into a storage tank, the slurry is sprayed into a drum containing 100 kg of granular fertilizer by using a water pump nozzle, the rotation speed of the drum is 15 r / min, and the Bacillus velezensis coated granular fertilizer is obtained after drum coating.

[0141] Example 5

[0142] Example 5 provides a composite Bacillus coated granular fertilizer.

[0143] The preparation method of the above-mentioned bacillus complex coated granular fertilizer is as follows: 5 kg of bacillus complex slurry (containing 200 billion cfu / mL) is loaded into a storage tank, and the slurry is sprayed into a drum containing 100 kg of granular fertilizer by using a water pump nozzle, the rotating speed of the drum is 15 r / min, and the bacillus complex coated granular fertilizer is obtained after coating by the drum.

[0144] The above-mentioned bacillus complex slurry is a mixture of bacillus subtilis, bacillus velezensis and bacillus amyloliquefaciens. The bacterial content of bacillus subtilis, bacillus velezensis and bacillus amyloliquefaciens is 200 billion cfu / mL, and the weight ratio of bacillus subtilis, bacillus velezensis and bacillus amyloliquefaciens is 1:2:1.

[0145] Comparative Example 5

[0146] Comparative Example 5 provides a bacillus velezensis coated granular fertilizer.

[0147] The above-mentioned comparative example and Example 2 are different in that the rotating speed of the drum in the preparation method of the bacillus velezensis coated granular fertilizer; the rotating speed of the drum in Comparative Example 5 is 5 r / min.

[0148] Comparative Example 6

[0149] Comparative Example 6 provides a bacillus velezensis coated granular fertilizer.

[0150] The above-mentioned comparative example and Example 2 are different in that the rotating speed of the drum in the preparation method of the bacillus velezensis coated granular fertilizer; the rotating speed of the drum in Comparative Example 6 is 20 r / min.

[0151] Comparative Example 7

[0152] Comparative Example 7 provides a bacillus velezensis coated granular fertilizer.

[0153] The above-mentioned bacillus velezensis coated granular fertilizer is a powder coated granular fertilizer. The preparation method of the bacillus velezensis coated granular fertilizer is as follows: the bacillus velezensis fermentation broth provided in Preparation Example 2 is separated in a butterfly centrifuge, the centrifugal speed is set to 5000 r / min, the centrifugal time is 250 s, the displacement is 0.3 L, and the bacillus velezensis slurry is obtained after centrifugation; then the bacillus velezensis slurry 5 kg (containing 200 billion cfu / mL) is dried by using a high-temperature spray dryer to obtain bacillus velezensis powder; then the bacillus velezensis powder is loaded into a storage tank, and the slurry is sprayed into a roller drum containing 100 kg of granular fertilizer by using a water pump nozzle, the roller drum rotates at a speed of 10-15 r / min, and an appropriate amount of water is added to make the bacillus velezensis powder completely coated on the surface of the granular fertilizer particles, and the powder type bacillus velezensis coated granular fertilizer is obtained after the roller coating.

[0154] Comparative Example 8

[0155] Comparative Example 8 provides a bacillus velezensis coated granular fertilizer.

[0156] The preparation method of the above-mentioned bacillus velezensis coated granular fertilizer is as follows: the bacillus velezensis fermentation broth provided in Preparation Example 2 is pumped into a mixed sedimentation tank, then the temperature of the fermentation tank is increased to 38℃, and the pH is adjusted to 5; then 0.3% of the polyphosphorus iron sulfate aqueous solution is pumped into the mixing tank, stirred at a speed of 160 r / min for 5 min, and then treated by standing; after the fermentation broth is clarified, the fermentation broth is pumped into a centrifuge and concentrated at 8000 r / min to obtain a slurry; then the slurry is pumped into a mixing tank, and fertilizer particles are added to the mixing tank to adsorb the concentrated fermentation broth and the fertilizer particles to obtain a mixture; finally, the mixture is sent to a flash dryer for medium-temperature drying treatment, and a powder type bacillus velezensis coated granular fertilizer is obtained.

[0157] Viable cell count detection

[0158] The viable cell count of the bacillus velezensis coated granular fertilizer obtained in Examples 1-5 and Comparative Examples 5-8 is detected, and the detection results are shown in Table 3.

[0159] The detection method of the viable cell count of the bacillus velezensis coated granular fertilizer is as follows: 10 g of the bacillus velezensis coated granular fertilizer is crushed and dissolved in 100 mL of sterile water, and then the viable cell count is counted according to the method of GB20287-2006, which is recorded as the initial viable cell count; then the bacillus velezensis coated granular fertilizer is stored for 1 year, and the viable cell count is counted again by using the same method, which is recorded as the viable cell count after 1 year; the viable cell count reduction rate is calculated by using the initial viable cell count and the viable cell count after 1 year, and the calculation formula is as follows:

[0160] Viable cell count reduction rate = (viable cell count after 1 year - initial viable cell count) / initial viable cell count

[0161] Table 3: Test results of viable cell count of bacillus encapsulated granular fertilizer obtained in Examples 1-5 and Comparative Examples 5-8

[0162]

[0163]

[0164] According to the test results of Examples 1-5 and Comparative Examples 5-8, it can be seen that the production process of bacillus encapsulated granular fertilizer provided in Examples 1-5 can prepare bacillus subtilis encapsulated granular fertilizer, bacillus velezensis encapsulated granular fertilizer, bacillus amyloliquefaciens encapsulated granular fertilizer and composite bacillus encapsulated granular fertilizer. The initial viable cell count of the above-mentioned bacillus encapsulated granular fertilizer is 9.4-9.6 billion cfu / mL, and after 1 year of storage, the viable cell count reduction rate is 1.04-6.31%. Although the initial viable cell count of the bacillus velezensis encapsulated granular fertilizer provided in Comparative Example 8 is 9.1 billion cfu / mL, after 1 year, the viable cell count reduction rate is as high as 9.89%. Therefore, it shows that the bacillus encapsulated granular fertilizer obtained by using the production process of bacillus encapsulated granular fertilizer provided in the present application has good stability, slow viable cell count reduction and long shelf life.

[0165] According to the test results of Comparative Examples 1-3, it can be seen that the viable cell count reduction rates of bacillus subtilis encapsulated granular fertilizer and bacillus amyloliquefaciens encapsulated granular fertilizer obtained in Example 1 and Example 3 after 1 year are 6.31% and 4.21% respectively. The viable cell count reduction rate of bacillus velezensis encapsulated granular fertilizer provided in Example 2 after 1 year is only 2.12%, which shows that the bacillus encapsulated granular fertilizer prepared by using bacillus velezensis has better stability, slower viable cell count reduction and longer shelf life.

[0166] According to the test results of Comparative Example 2, Example 4 and Comparative Examples 5-6, it can be seen that with the increase of the rotating speed of the roller, the initial bacterial content of bacillus velezensis encapsulated granular fertilizer shows a trend of first increasing and then decreasing. Although the viable cell count reduction rate is basically unchanged after 1 year of storage, the viable cell count reduction rates of bacillus velezensis encapsulated granular fertilizer provided in Comparative Examples 5-6 are relatively high. Therefore, it shows that by controlling the rotating speed of the roller between 10-15 r / min, the bacillus encapsulated granular fertilizer can have a higher viable cell count, thereby obtaining a longer shelf life.

[0167] The detection results of Comparative Example 2 and Comparative Example 7 show that the initial viable bacterial count of the Bacillus velezensis liquid coated granular fertilizer provided in Example 2 is obviously higher than that of the Bacillus velezensis powder coated granular fertilizer provided in Comparative Example 7, and the viable bacterial count of the Bacillus velezensis powder coated granular fertilizer provided in Comparative Example 7 decreases by 8.64% after being stored for 1 year. Therefore, it is proved that the preparation process of the Bacillus coated granular fertilizer provided in the present application can prepare a Bacillus liquid coated granular fertilizer, which has high viable bacterial count, slow viable bacterial count decrease and long shelf life.

[0168] The detection results of Comparative Examples 1-3 and Example 5 show that the initial viable bacterial count of the composite Bacillus coated granular fertilizer obtained in Example 5 is 950 million cfu / mL, the viable bacterial count is 900 million cfu / mL after being stored for 1 year, and the viable bacterial count decreases by 5.26%. It is proved that the composite Bacillus coated granular fertilizer with high viable bacterial count and long shelf life can also be prepared by mixing Bacillus subtilis, Bacillus velezensis and Bacillus amyloliquefaciens.

[0169] Application Examples

[0170] Application Examples 1-4

[0171] Application Examples 1-4 respectively provide a cultivation method of a tomato plant.

[0172] The difference between the above application examples lies in the source of the Bacillus coated granular fertilizer in the soil in the cultivation method of the tomato plant. The Bacillus coated granular fertilizer in Application Examples 1-3 respectively comes from Examples 1-3, and the Bacillus coated granular fertilizer in Application Example 4 respectively comes from Example 5.

[0173] The cultivation method of the tomato plant provided in Application Example 1 is specifically as follows: take a plastic pot, fill it with 2 kg of soil, transplant the tomato plant into the plastic pot, and after the tomato plant is planted for 8 days, uniformly apply 1 g of the Bacillus coated granular fertilizer provided in Example 1 to the soil in the plastic pot, and then perform field management on the plastic pot every day.

[0174] Application Example 5

[0175] Application Example 5 provides a cultivation method of a tomato plant.

[0176] The difference between the above application examples and Application Example 2 lies in that no Bacillus coated granular fertilizer is applied to the soil.

[0177] Performance detection test

[0178] The application performance of the Bacillus coated granular fertilizer provided in Examples 1-3 and Example 5 of the present application is detected, and the results are shown in Table 4. The specific method is as follows:

[0179] Test method: according to the method provided in application examples 1-5 to cultivate tomato plants, after the tomato plants grow for 30 days, use a tape measure to measure the plant height L of the tomato plants; use a vernier caliper to measure the stem diameter Q of the tomato plants, the measurement position of the stem diameter is 1 cm from the stem base.

[0180] Calculate the growth rate of the plant height L1 of the tomato plants in application examples 1-4 compared to the plant height L0 of the tomato plants in application example 5 (blank control group); and the growth rate of the stem diameter Q1 of the tomato plants in application examples 1-4 compared to the stem diameter Q0 of the tomato plants in application example 5 (blank control group). The calculation formula is as follows:

[0181] Plant height growth rate = (L1-L0) / L0 x 100%

[0182] Stem diameter growth rate = (Q1-Q0) / Q0 x 100%

[0183] Wherein, L1 is the plant height of the tomato plants in application examples 1-4, L0 is the plant height of the tomato plants in application example 5; Q1 is the stem diameter of the tomato plants in application examples 1-4, Q0 is the stem diameter of the tomato plants in application example 5

[0184] Table 4: Test results of the plant height and stem diameter of the tomato plants obtained in application examples 1-5

[0185]

[0186]

[0187] According to the above test results, it can be seen that the bacillus encapsulated granular fertilizer provided in application examples 1-3 and example 5 can promote the growth of tomato plants, thereby significantly increasing the plant height and diameter of the tomato plants.

[0188] Although the present application has been described in detail in the foregoing description and specific embodiments, it is clear that modifications and improvements can be made to the present application without departing from the spirit of the present application, which will be apparent to those skilled in the art. Therefore, these modifications and improvements made on the basis of the spirit of the present application are within the scope of the present application.

Claims

1. A Bacillus encapsulated granular fertilizer, characterized in that, The bacillus encapsulated granular fertilizer comprises bacterial slurry and fertilizer granules; the bacterial slurry is wrapped on the outer surface of the fertilizer granules; the bacterial content of the bacterial slurry is 20-50 billion cfu / mL, and the viscosity is 8000-10000 mPa; The production process of the bacillus encapsulated granular fertilizer comprises the following steps: (1) Seed liquid fermentation: first, culture the seed liquid, then ferment and culture the seed liquid to obtain a fermentation liquor; (2) Centrifugation: centrifuge the fermentation liquor in a centrifugal device to obtain bacterial slurry; the centrifugation rate is 5000-6000 r / min, and the centrifugation time is 250-300 s; (3) Encapsulation: use an encapsulation device to wrap the bacterial slurry on the outer surface of the fertilizer granules to obtain bacillus encapsulated granular fertilizer; the rotation speed of the encapsulation device is 10-15 r / min.

2. The Bacillus encapsulated granular fertilizer of claim 1, wherein, The bacterial content of the bacterial slurry is 250-350 billion cfu / mL, and the viscosity is 9000-9500 mPa·s.

3. The Bacillus encapsulated granular fertilizer of claim 1, wherein, The bacillus is selected from Bacillus subtilis, Bacillus velezensis and Bacillus amyloliquefaciens.

4. The Bacillus encapsulated granular fertilizer of claim 3, wherein, The bacillus is a mixture of Bacillus subtilis, Bacillus velezensis and Bacillus amyloliquefaciens.

5. The Bacillus encapsulated granular fertilizer of claim 1, wherein, The fermentation culture uses LB medium and / or full water-soluble medium.

6. The bacillus encapsulated granular fertilizer according to any one of claims 1-5 is applied to soil.

7. Use of the Bacillus encapsulated granular fertilizer according to claim 6 in soil, characterized in that, The weight ratio of the bacillus encapsulated granular fertilizer to soil is (0.8-1.2):2000.

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