Agricultural glycinin extract and application thereof in agriculture

Abstract

The invention relates to a glycinin extract for agriculture and application thereof in agriculture, which is characterized by comprising the steps of globulin extraction and microorganism extraction, the microorganism extracted from the microorganism is bacillus N311 which is preserved in the China Center for Type Culture Collection on October 15, 2015, and the preservation number of the China Center for Type Culture Collection is CCTCC M 2015607; the bacillus N311 is bacillus velezensis, and the bacillus N311 is bacillus velezensis. The glycinin extracting solution prepared by the invention not only has better antibacterial property, but also can promote the growth of crops.

Description

TECHNICAL FIELD

[0001] The present application relates to a soybean protein extract for agriculture and its application in agriculture. BACKGROUND

[0002] The Bacillus from Antarctica N311 has been preserved in the China Center for Type Culture Collection on October 15, 2015, with the preservation center preservation number CCTCC M 2015607, and the patent number of the applied strain is 2015109992628, and the patent name is the Bacillus from Antarctica N311 and its application in preventing and treating plant pathogenic fungi. The supernatant prepared by the Bacillus from Antarctica N311 has good bacteriostasis.

[0003] Our company mainly engages in the research and development, production and import and export business of microbial preparations, water-soluble fertilizers and the like. Our company believes that the Antarctic has a special geographical environment, and the growth conditions of the strain are much worse than those in the inland, so it may have better prospects for application in agriculture. After the company's evaluation, the Bacillus from Antarctica N311 has good application prospects, and the company has the ability to transform the patents of scientific research institutions. After several negotiations, our company purchased the patent, and on December 27, 2023, the patent was transferred to our company. In order to further determine the functionality of the strain, our company applied for the identification of the strain, and the identification result was issued on March 4, 2025. The strain is Bacillus velezensis.

[0004] After one and a half years of research, our company further developed the new function of promoting crop growth of the extract prepared by the Bacillus from Antarctica N311 on the basis of retaining the bacteriostasis of the extract, and the application in crops has good yield-increasing effect.

[0005] At present, there is no method and product for further research on the Bacillus from Antarctica N311. SUMMARY

[0006] The present application provides a soybean protein extract for agriculture and its application in agriculture, which solves the technical problem of further developing the new function of the extract prepared by the Bacillus from Antarctica N311 on the basis of retaining the bacteriostasis of the extract, and the application in crops has good effect.

[0007] In order to solve the above technical problem, the present application adopts the following technical scheme:

[0008] A soybean globulin extract for agriculture, comprising globulin extraction and microbial extraction; the microorganism of the microbial extraction is Bacillus velezensis N311, which has been preserved in the China Center for Type Culture Collection on October 15, 2015, and the preservation center preservation number is CCTCC M 2015607; the strain was re-identified as Bacillus velezensis on March 4, 2025 by the School of Life Science and Technology of Huazhong Agricultural University and the Microbial Product Quality Inspection and Test Center of the Ministry of Agriculture and Rural Affairs (Wuhan).

[0009] The globulin extraction is carried out according to the following steps

[0010] 30-50 mesh soybean meal is weighed, and the weighed soybean meal is added to a buffer solution with a pH of 8-9, and extracted at a rotation speed of 120-220 r / min and a temperature of 30-50℃ for 2-4 h, filtered to obtain filtrate 1 and residue 1, and the obtained filtrate 1 is a globulin extract;

[0011] The residue 1 is dispersed by stirring at a rotation speed of 120-220 r / min, water is added, sterilization is performed, Bacillus velezensis N311 seed liquid is added, fermentation is carried out in a closed environment, sterilization and filtration are performed, and filtrate 2 and residue 2 are obtained, and the obtained filtrate 2 is a microbial extract;

[0012] The globulin extract and the microbial extract are combined to obtain a soybean globulin extract for agriculture;

[0013] The soybean globulin extract for agriculture is dried to a moisture content of less than 6% to obtain a soybean globulin extract powder for agriculture;

[0014] The soybean globulin extract for agriculture comprises a soybean globulin extract for agriculture and a soybean globulin extract powder for agriculture;

[0015] The mass ratio of the soybean meal to the buffer solution is 1:12-18;

[0016] The mass ratio of the residue 1 to water is 1:0.1-0.4;

[0017] The mass ratio of the Bacillus velezensis N311 seed liquid to the residue 1 is 1-4:96-99;

[0018] The oxygen content in the closed space during fermentation is maintained at 5%-15%;

[0019] The fermentation time is 72-96 h, the fermentation temperature is 28-30℃, and the fermentation stirring rotation speed is 120-220 r / min;

[0020] The sterilization temperature is 100-121℃, and the time is 20-30 min;

[0021] The dry material temperature is not more than 60 DEG C.

[0022] The bacillus N311 seed liquid is prepared according to the following steps,

[0023] The bacillus N311 is inoculated into the seed culture medium, and is cultured at 28-30 DEG C for 24-48 h to obtain activated bacillus N311;

[0024] The activated bacillus N311 is inoculated into the LB culture medium, and is cultured at 28-30 DEG C with 180-220 r / min oscillation for 48-60 h to obtain the bacillus N311 seed liquid;

[0025] The seed culture medium is: sodium chloride 10 g / L, tryptone 10 g / L, yeast extract 5 g / L, agar 20 g / L, and the rest is water, and is sterilized at 121 DEG C for 20 min;

[0026] The LB culture medium is: sodium chloride 10 g / L, tryptone 10 g / L, yeast extract 5 g / L, and the rest is water, and is sterilized at 121 DEG C for 20 min.

[0027] The application of the soybean protein extract for agriculture in agriculture, characterized in that the soybean protein extract for agriculture is used as a nutrient solution, and is applied to crops; or

[0028] The soybean protein extract for agriculture is used as a raw material in a fertilizer.

[0029] The nutrient solution is a solution obtained by diluting the soybean protein extract for agriculture with water at a ratio of 200-1500 times;

[0030] The soybean protein extract for agriculture is used as a raw material in a fertilizer, and the mass ratio of the soybean protein extract for agriculture to the fertilizer is 5-90:10-95.

[0031] The soybean protein extract for agriculture is used as a raw material in a fertilizer, and the material temperature is not more than 60 DEG C during drying.

[0032] The application has the following beneficial technical effects:

[0033] 1. The application is a further development of the function of the antifungal bacillus N311 from Antarctica, and belongs to a new application of an old product, and has obvious creativity.

[0034] 2. This application includes globulin extraction and microbial extraction. Experiments have shown that microbial extraction alone is inferior to globulin extraction and microbial extraction in terms of both antibacterial properties and crop growth promotion. The possible reason is that Bacillus N311 extraction degrades soybean globulin, thus affecting the effect of use.

[0035] 3. During the drying process of this application, the material temperature shall not exceed 60℃. Exceeding 60℃ will affect the performance and may be related to protein denaturation caused by high temperature. Detailed Implementation

[0036] To better understand the above technical solutions, the technical solutions of the present invention will be described in detail below with reference to preferred examples.

[0037] Example 1

[0038] A soybean globulin extract for agricultural use includes globulin extraction and microbial extraction; the microorganism used for microbial extraction is Bacillus N311, which was deposited at the China Center for Type Culture Collection on October 15, 2015, with accession number CCTCC M 2015607; this strain was identified as Bacillus belyes by the College of Life Science and Technology of Huazhong Agricultural University and the Microbial Product Quality Inspection and Testing Center of the Ministry of Agriculture and Rural Affairs (Wuhan) on March 4, 2025.

[0039] The globulin extraction is performed according to the following steps.

[0040] Weigh out 40-mesh soybean meal and add it to a Tris-HCl buffer solution with a pH of 8.50. Extract for 3 hours at a speed of 180 r / min and a temperature of 40℃. Filter to obtain filtrate 1 and residue 1. Filtrate 1 is the globulin extract.

[0041] Stirring at 180 r / min, breaking up filter residue 1, adding water, sterilizing, adding Bacillus N311 seed liquid, fermenting in a closed environment, sterilizing and filtering to obtain filtrate 2 and filter residue 2, the obtained filtrate 2 is the microbial extract;

[0042] Combining the globulin extract and the microbial extract yields a soybean globulin extract suitable for agricultural use.

[0043] Soybean globulin extract for agricultural use is dried to a moisture content of 5% to obtain soybean globulin extract powder for agricultural use.

[0044] The drying process is spray drying at a temperature of 58°C.

[0045] The mass ratio of soybean meal to buffer solution is 1:14;

[0046] The mass ratio of the residue 1 and water is 1:0.2;

[0047] The mass ratio of the bacillus N311 seed liquid and the residue 1 is 2:98;

[0048] The oxygen content in the closed space is maintained at 12% during the fermentation process;

[0049] The fermentation time is 84h, the fermentation temperature is 28℃, and the fermentation stirring speed is 180r / min;

[0050] The sterilization temperature is 121℃, and the time is 20min.

[0051] The bacillus N311 seed liquid is prepared according to the following steps,

[0052] The bacillus N311 is inoculated into the seed culture medium, cultured at 28℃ for 24h, activated, and the activated bacillus N311 is obtained;

[0053] The activated bacillus N311 is inoculated into the LB culture medium, and cultured at 28℃ with 180r / min shaking for 48h, which is the bacillus N311 seed liquid;

[0054] The seed culture medium is: sodium chloride 10g / L, tryptone 10g / L, yeast extract 5g / L, agar 20g / L, and the rest is water, sterilized at 121℃ for 20min;

[0055] The LB culture medium is: sodium chloride 10g / L, tryptone 10g / L, yeast extract 5g / L, and the rest is water, sterilized at 121℃ for 20min.

[0056] The application of the soybean protein extract for agriculture in agriculture, the soybean protein extract for agriculture is used as a nutrient solution, applied to crops; or

[0057] The soybean protein extract for agriculture is used as a raw material in fertilizer.

[0058] The nutrient solution is a solution obtained by diluting the soybean protein extract for agriculture 200 times with water; or the nutrient solution is a solution obtained by diluting the soybean protein extract powder for agriculture 1200 times with water;

[0059] The soybean protein extract for agriculture is used as a raw material in fertilizer, and the mass ratio of the soybean protein extract for agriculture and urea is 80:20; or

[0060] The soybean protein extract for agriculture is used as a raw material in fertilizer, and the mass ratio of the soybean protein extract powder for agriculture and urea is 10:90;

[0061] Tris-HCl buffer solution with pH 8.50 is configured as follows: 25 ml of 0.2 mol / L Tris and 15 ml of 0.1N HCl.

[0062] Example 2

[0063] A soybean globulin extract for agriculture, comprising globulin extraction and microbial extraction; the microorganism of the microbial extraction is Bacillus N311, which has been preserved in the China Center for Type Culture Collection on October 15, 2015, with the preservation center preservation number CCTCC M 2015607; the strain was re-identified as Bacillus velezensis by the College of Life Science and Technology, Huazhong Agricultural University and the Microbial Product Quality Inspection and Test Center of the Ministry of Agriculture and Rural Affairs (Wuhan) on March 4, 2025.

[0064] The globulin extraction is carried out according to the following steps

[0065] 40-mesh soybean meal is weighed, and the weighed soybean meal is added to a Tris-HCl buffer solution with pH 8.23, and extracted at a rotation speed of 180 r / min and a temperature of 45℃ for 3 h, filtered to obtain filtrate 1 and residue 1, and the obtained filtrate 1 is a globulin extract;

[0066] The residue 1 is dispersed by stirring at a rotation speed of 180 r / min, water is added, sterilized, Bacillus N311 seed liquid is added, and fermented in a closed environment, sterilized, filtered to obtain filtrate 2 and residue 2, and the obtained filtrate 2 is a microbial extract;

[0067] The globulin extract and the microbial extract are combined to obtain a soybean globulin extract for agriculture;

[0068] The soybean globulin extract for agriculture is dried to a moisture content of 5% to obtain a soybean globulin extract powder for agriculture;

[0069] The drying is spray drying, and the drying temperature is 58℃;

[0070] The mass ratio of the soybean meal to the buffer solution is 1:16;

[0071] The mass ratio of the residue 1 to water is 1:0.3;

[0072] The mass ratio of the Bacillus N311 seed liquid to the residue 1 is 3:97;

[0073] The oxygen content in the closed space during the fermentation process is maintained at 14%;

[0074] The fermentation time is 84 h, the fermentation temperature is 28℃, and the fermentation stirring rotation speed is 180 r / min;

[0075] The sterilization temperature is 121℃ and the time is 20 minutes.

[0076] The bacillus N311 seed liquid is prepared according to the following steps,

[0077] The bacillus N311 is inoculated into the seed culture medium, cultured at 30℃ for 24 hours to activate, and the activated bacillus N311 is obtained;

[0078] The activated bacillus N311 is inoculated into the LB culture medium, and cultured at 30℃ with 180r / min shaking for 48 hours to obtain the bacillus N311 seed liquid;

[0079] The seed culture medium is: sodium chloride 10g / L, tryptone 10g / L, yeast extract 5g / L, agar 20g / L, and the rest is water, sterilized at 121℃ for 20 minutes;

[0080] The LB culture medium is: sodium chloride 10g / L, tryptone 10g / L, yeast extract 5g / L, and the rest is water, sterilized at 121℃ for 20 minutes.

[0081] The application of the soybean protein extract for agriculture in agriculture uses the soybean protein extract for agriculture as a nutrient solution and applies it to crops; or

[0082] The soybean protein extract for agriculture is used as a raw material in fertilizer.

[0083] The nutrient solution is a solution obtained by diluting the soybean protein extract solution for agriculture 200 times with water; or the nutrient solution is a solution obtained by diluting the soybean protein extract powder for agriculture 1200 times with water.

[0084] The soybean protein extract for agriculture is used as a raw material in fertilizer, and the mass ratio of the soybean protein extract solution for agriculture, urea and potassium dihydrogen phosphate is 60:32:8.

[0085] The soybean protein extract solution for agriculture is used as a raw material in fertilizer, and the mass ratio of the soybean protein extract powder for agriculture, urea and potassium dihydrogen phosphate is 8:52:40.

[0086] The Tris-HCl buffer solution with pH 8.23 is prepared by mixing 25ml of 0.2mol / L Tris and 22.5ml of 0.1N HCl.

[0087] Example 3

[0088] A soybean globulin extract for agriculture, comprising globulin extraction and microbial extraction; the microorganism of the microbial extraction is Bacillus velezensis N311, which has been preserved in the China Center for Type Culture Collection on October 15, 2015, and the preservation center preservation number is CCTCC M 2015607; the strain was re-identified as Bacillus velezensis on March 4, 2025 by the School of Life Science and Technology of Huazhong Agricultural University and the Microbial Product Quality Inspection and Test Center of the Ministry of Agriculture and Rural Affairs (Wuhan).

[0089] The globulin extraction is carried out according to the following steps

[0090] 40-mesh soybean meal is weighed, and the weighed soybean meal is added to a Tris-HCl buffer solution with a pH of 8.74, and extracted at a rotation speed of 200 r / min and a temperature of 47℃ for 4 h, filtered to obtain filtrate 1 and residue 1, and the obtained filtrate 1 is a globulin extract;

[0091] The residue 1 is dispersed by stirring at a rotation speed of 200 r / min, water is added, sterilization is performed, Bacillus velezensis N311 seed liquid is added, fermentation is performed in a closed environment, sterilization, filtration are performed, and filtrate 2 and residue 2 are obtained, and the obtained filtrate 2 is a microbial extract;

[0092] The globulin extract and the microbial extract are combined to obtain a soybean globulin extract for agriculture;

[0093] The soybean globulin extract for agriculture is dried to a moisture content of 5% to obtain a soybean globulin extract powder for agriculture;

[0094] The drying is spray drying, and the drying temperature is 56℃;

[0095] The mass ratio of the soybean meal to the buffer solution is 1:17;

[0096] The mass ratio of the residue 1 to water is 1:0.4;

[0097] The mass ratio of the Bacillus velezensis N311 seed liquid to the residue 1 is 4:96;

[0098] The oxygen content in the closed space during fermentation is maintained at 12%;

[0099] The fermentation time is 84 h, the fermentation temperature is 28℃, and the fermentation stirring rotation speed is 200 r / min;

[0100] The sterilization temperature is 121℃, and the time is 20 min.

[0101] The Bacillus velezensis N311 seed liquid is prepared according to the following steps,

[0102] The bacillus N311 is inoculated into a seed culture medium, and cultivated at 30℃ for 24 hours to activate the bacillus N311;

[0103] The activated bacillus N311 is inoculated into an LB culture medium, and cultivated at 30℃ with 200r / min oscillation for 48 hours to obtain the bacillus N311 seed liquid;

[0104] The seed culture medium is: sodium chloride 10g / L, tryptone 10g / L, yeast extract 5g / L, agar 20g / L, and the rest is water, sterilized at 121℃ for 20 minutes;

[0105] The LB culture medium is: sodium chloride 10g / L, tryptone 10g / L, yeast extract 5g / L, and the rest is water, sterilized at 121℃ for 20 minutes.

[0106] The application of the soybean protein extract for agriculture in agriculture uses the soybean protein extract for agriculture as a nutrient solution, and applies it to crops; or

[0107] The soybean protein extract for agriculture is used as raw material in fertilizer.

[0108] The nutrient solution is a solution obtained by diluting the soybean protein extract for agriculture 200 times with water; or a solution obtained by diluting the soybean protein extract powder for agriculture 1200 times with water.

[0109] The soybean protein extract for agriculture is used as raw material in fertilizer, and the mass ratio of the soybean protein extract for agriculture, urea and potassium dihydrogen phosphate is 60:32:8.

[0110] The soybean protein extract for agriculture is used as raw material in fertilizer, and the mass ratio of the soybean protein extract powder for agriculture, urea and potassium dihydrogen phosphate is 8:52:40.

[0111] The pH8.74 Tris-HCl buffer solution is configured as follows: 25ml of 0.2mol / L Tris and 10ml of 0.1N HCl.

[0112] Example 4

[0113] A soybean globulin extract for agriculture, comprising globulin extraction and microbial extraction; the microorganism of the microbial extraction is Bacillus N311, which has been preserved in the China Center for Type Culture Collection on October 15, 2015, and the preservation center preservation number is CCTCC M 2015607; the strain was re-identified as Bacillus velezensis by the College of Life Science and Technology, Huazhong Agricultural University and the Microbial Product Quality Inspection and Test Center of the Ministry of Agriculture and Rural Affairs (Wuhan) on March 4, 2025.

[0114] The globulin extraction is carried out according to the following steps

[0115] 40-mesh soybean meal is weighed, and the weighed soybean meal is added to a barbituric acid buffer solution with a pH of 8.6, and extracted at a rotation speed of 200 r / min and a temperature of 47℃ for 4 h, filtered to obtain filtrate 1 and residue 1, and the obtained filtrate 1 is a globulin extract;

[0116] The residue 1 is dispersed by stirring at a rotation speed of 200 r / min, water is added, sterilized, and Bacillus N311 seed liquid is added, and fermented in a closed environment, sterilized, filtered to obtain filtrate 2 and residue 2, and the obtained filtrate 2 is a microbial extract;

[0117] The globulin extract and the microbial extract are combined to obtain a soybean globulin extract for agriculture;

[0118] The soybean globulin extract for agriculture is dried to a moisture content of 5% to obtain a soybean globulin extract powder for agriculture;

[0119] The drying is spray drying, and the drying temperature is 55℃;

[0120] The mass ratio of the soybean meal to the buffer solution is 1:16;

[0121] The mass ratio of the residue 1 to water is 1:0.3;

[0122] The mass ratio of the Bacillus N311 seed liquid to the residue 1 is 4:96;

[0123] The oxygen content in the closed space during the fermentation process is maintained at 15%;

[0124] The fermentation time is 84 h, the fermentation temperature is 28℃, and the fermentation stirring rotation speed is 200 r / min;

[0125] The sterilization temperature is 121℃, and the time is 20 min.

[0126] The Bacillus N311 seed liquid is prepared according to the following steps,

[0127] The Bacillus sp. N311 is inoculated into a seed culture medium, and cultivated at 30℃ for 24 hours to activate the Bacillus sp. N311, and the activated Bacillus sp. N311 is inoculated into an LB culture medium, and cultivated at 30℃ with 200r / min oscillation for 48 hours to obtain the seed liquid of the Bacillus sp. N311;

[0128] The LB culture medium is: sodium chloride 10g / L, tryptone 10g / L, yeast extract 5g / L, and the rest is water, and sterilized at 121℃ for 20 minutes.

[0129] The LB culture medium is: sodium chloride 10g / L, tryptone 10g / L, yeast extract 5g / L, and the rest is water, and sterilized at 121℃ for 20 minutes.

[0130] The LB culture medium is: sodium chloride 10g / L, tryptone 10g / L, yeast extract 5g / L, and the rest is water, and sterilized at 121℃ for 20 minutes.

[0131] The soybean protein extract for agriculture is applied to the fertilizer as a raw material.

[0132] The soybean protein extract for agriculture is applied to the fertilizer as a raw material.

[0133] The nutrient solution is a solution obtained by diluting the soybean protein extract solution for agriculture by 200 times with water, or a solution obtained by diluting the soybean protein extract powder for agriculture by 1200 times with water.

[0134] The soybean protein extract for agriculture is applied to the fertilizer as a raw material.

[0135] The soybean protein extract for agriculture is applied to the fertilizer as a raw material.

[0136] The pH 8.6 barbitone buffer solution is configured as follows: barbitone 1.84g, sodium barbitone 10.3g, and distilled water to 1000.00ml.

[0137] The beneficial effects of the present application are further illustrated below in combination with test data:

[0138] Experiment 1

[0139] 1.1 Test site: Shandong Tudakuer Fertilizer Co., Ltd.

[0140] 1.2 Experimental detection: inhibition diameter (mm), root activity (mg / g.h), dry root weight (g), and aboveground weight (g).

[0141] 1.3 Test materials: Comparative Example 1 (identical to Example 1 except that the globulin extract prepared in Example 1 was used), Comparative Example 2 (identical to Example 1 except that the microbial extract prepared in Example 1 was used), Comparative Example 3 (identical to Example 1 except that Bacillus velezensis was purchased from China Center for Type Culture Collection, with the preservation number CCTCC AB 2016194, and the strain was derived from the soil adjacent to the paddy field at Xiaoshanjiao, Lichuan, Hubei, and had a broad antibacterial property), Comparative Example 4 (identical to Example 1 except that Bacillus velezensis was purchased from China Center for Type Culture Collection, with the preservation number CCTCC AB 205354, and the strain was derived from sandy soil in Xinjiang), and the globulin extracts prepared in Examples 1 to 4 for agricultural use.

[0142] 1.4 Test method: Lettuce was used for the plant test, and gravel culture was used, and the gravel was sterilized at 160°C for 2h before use. The culture pots had a mouth diameter of 38cm, a bottom diameter of 30cm, and a height of 40.5cm, and two plants were planted in each pot. On the 7th day after transplanting, each pot was irrigated with each treatment diluted 300 times with water, a total of 4 times, and sampling was performed on the 31st day for detection. Two obviously large and two obviously small plants were removed from each treatment, and 20 pots were planted for each treatment.

[0143] 1.5 Detection method:

[0144] Inhibition zone diameter: the bacteria that had been incubated for 24h were taken twice with a inoculation loop, and then uniformly smeared on the surface of the culture medium with a coating rod, spotted on a 6mm wide filter paper, and placed at 37°C for one day. The diameter (mm) of the inhibition zone was measured with a vernier caliper. The selected bacteria for the experiment were Fusarium graminearum.

[0145] Dry root weight and aboveground weight were measured by weighing, washing, and drying in a 60°C oven until a constant weight was obtained, with a constant weight difference of less than 0.02g between two consecutive measurements.

[0146] Root activity was measured by the α-naphthylamine method.

[0147] The treatments in this experiment were different, and the other treatments were consistent.

[0148] 2 Results and analysis

[0149] Inhibition zone diameter (mm), root activity (mg / g.h), dry root weight (g), and aboveground weight (g) are shown in Table 1

[0150] Table 1

[0151] Inhibition zone diameter (mm) Root activity (mg / g.h) Dry root weight (g) Above ground weight (g) Comparative 1 6.73 0.28 0.28 1.78 Comparative 2 6.24 0.29 0.29 1.82 Comparative 3 7.43 0.31 0.31 1.90 Comparative 4 7.21 0.32 0.31 1.92 Example 1 8.12 0.36 0.34 1.96 Example 2 8.25 0.38 0.38 2.02 Example 3 8.17 0.35 0.35 1.98 Example 4 8.04 0.33 0.32 1.93

[0152] From the comparison of the data of Comparative Example 1, Comparative Example 2 and Example 1 in Table 1, it can be seen that the soybean globulin extract for agriculture prepared by combining the globulin extract and the microbial extract has a much better effect than the globulin extract and the microbial extract.

[0153] From the comparison of the data of Comparative Example 3, Comparative Example 4 and Example 1, it can be seen that the effect of Example 1 prepared in the present application is more obvious in terms of antibacterial property, but there is no obvious advantage in promoting crop growth.

[0154] Experiment Two

[0155] 1.1 Test site: Daliuzhuang, Jining Yutai County; Lettuce variety: Rosa.

[0156] 1.2 Experimental detection: yield (kg / acre).

[0157] 1.3 Test materials: Comparative Example 1 (except that the globulin extract prepared in Example 1 is used, other conditions are the same as those in Example 1), Comparative Example 2 (except that the microbial extract prepared in Example 1 is used, other conditions are the same as those in Example 1), Comparative Example 3 (except that Bacillus velezensis purchased from China Center for Type Culture Collection, preservation number CCTCC AB 2016194 is used, other conditions are the same as those in Example 1, the strain is derived from the soil adjacent to the paddy field at the foot of Xiaoshan in Lichuan, Hubei, and has a broad antibacterial property), Comparative Example 4 (except that Bacillus velezensis purchased from China Center for Type Culture Collection, preservation number CCTCC AB 205354 is used, other conditions are the same as those in Example 1, the strain is derived from sandy soil in Xinjiang), and the soybean globulin extract for agriculture prepared in Examples 1 to 4.

[0158] 1.4 Test method: Two acres of lettuce gardens with similar plots were divided into 8 subplots, each with an area of 150 square meters. The plant spacing of lettuce was 30*30 cm. Drip irrigation was used, each time for 1.5 h, and the nutrient solution of each treatment was diluted by 300 times. The base fertilizer for each subplot was 15-15-15 potassium sulfate compound fertilizer, and the dosage was 10 kg per subplot.

[0159] 1.5 Detection method: each treatment was harvested uniformly, and each subplot was weighed separately.

[0160] The treatments in this experiment were different, and other treatments were consistent.

[0161] 2 Results and analysis

[0162] Yield (kg / acre), see Table 2

[0163] Table 2

[0164] Yield (kg / acre) Comparative 1 2017.4 Comparative 2 2038.6 Comparative 3 2115.1 Comparative 4 2179.4 Example 1 2204.5 Example 2 2297.1 Example 3 2247.1 Example 4 2189.3

[0165] As can be seen from the data of Table 1 and Table 2, the data of Comparative, Comparative 2 and Example 1 are compared, the soybean globulin extract for agriculture prepared by combining the globulin extract and the microbial extract is much better than the globulin extract and the microbial extract. While the difference between Comparative 3, Comparative 4 and Example 1 is not obvious in promoting crop growth in Table 1, but there is a significant gap in yield performance in Table 2, which may be due to the influence of bacteriostasis, high bacteriostasis can make the plant grow better and healthier, thereby increasing the yield, that is, Bacillus N311 has obvious advantages compared with ordinary strains.

Claims

1. A soy glycinin extract for use in agriculture, characterized in that, The method comprises globulin extraction and microbial extraction; the microorganism of the microbial extraction is Bacillus velezensis N311, which was preserved in the China Center for Type Culture Collection on October 15, 2015, and the preservation center preservation number is CCTCC M 2015607; the strain was re-identified as Bacillus velezensis on March 4, 2025 by the College of Life Science and Technology of Huazhong Agricultural University and the Microbial Product Quality Inspection and Test Center of the Ministry of Agriculture and Rural Affairs (Wuhan).

2. The glycinin extract for agricultural use according to claim 1, characterized by, The globulin extraction is performed according to the following steps 30-50 mesh soybean meal is weighed, and the weighed soybean meal is added into a buffer solution with a pH of 8-9, and then extracted at a rotation speed of 120-220 r / min and a temperature of 30-50 DEG C for 2-4 h, filtered to obtain filtrate 1 and residue 1, and the obtained filtrate 1 is a globulin extract; The residue 1 is dispersed by stirring at a rotation speed of 120-220 r / min, water is added, sterilization is performed, Bacillus velezensis N311 seed liquid is added, fermentation is performed in a closed environment, sterilization and filtration are performed to obtain filtrate 2 and residue 2, and the obtained filtrate 2 is a microbial extract; The globulin extract and the microbial extract are combined to obtain a soybean globulin extract for agriculture; The soybean globulin extract for agriculture is dried to a water content of less than 6% to obtain a soybean globulin extract powder for agriculture; The soybean globulin extract for agriculture comprises the soybean globulin extract liquid for agriculture and the soybean globulin extract powder for agriculture; The mass ratio of the soybean meal to the buffer solution is 1:12-18; The mass ratio of the residue 1 to water is 1:0.1-0.4; The mass ratio of the Bacillus velezensis N311 seed liquid to the residue 1 is 1-4:96-99; The oxygen content in the closed space is maintained at 5%-15% during the fermentation; The fermentation time is 72-96 h, the fermentation temperature is 28-30 DEG C, and the fermentation stirring rotation speed is 120-220 r / min; The sterilization temperature is 100-121 DEG C, and the time is 20-30 min; The drying material temperature is not higher than 60 DEG C.

3. The glycinin extract for agricultural use according to claim 1, characterized by, The Bacillus velezensis N311 seed liquid is prepared according to the following steps Bacillus velezensis N311 is inoculated into a seed culture medium, and then cultured at 28-30 DEG C for 24-48 h to obtain activated Bacillus velezensis N311; The activated Bacillus velezensis N311 is inoculated into an LB culture medium, and then cultured at 28-30 DEG C with oscillation at 180-220 r / min for 48-60 h to obtain the Bacillus velezensis N311 seed liquid; The seed culture medium is: sodium chloride 10 g / L, tryptone 10 g / L, yeast extract 5 g / L, agar 20 g / L, and the balance is water, and high-pressure sterilization is performed at 121 DEG C for 20 min; The LB culture medium is: sodium chloride 10 g / L, tryptone 10 g / L, yeast extract 5 g / L, and the balance is water, and high-pressure sterilization is performed at 121 DEG C for 20 min.

4. Use of a glycinin extract prepared according to any one of claims 1 to 3 for agriculture, characterized in that, The soybean globulin extract for agriculture is used as a nutrient solution and applied to crops; or The soybean globulin extract for agriculture is used as a raw material and applied to a fertilizer.

5. Use of a soy glycinin extract prepared according to claim 4 for agriculture, characterized in that, The nutrient solution is a solution obtained by diluting a soybean protein isolate for agricultural use with water at a dilution ratio of 200 to 1500 times; The soybean protein isolate for agricultural use is used as a raw material for the fertilizer, and the mass ratio of the soybean protein isolate for agricultural use to the fertilizer is 5 to 90:10 to 95.

6. Use of a glycinin extract prepared according to claim 4 or 5 for agriculture, characterized in that, The soybean protein isolate for agricultural use is used as a raw material for the fertilizer, and the material temperature is not more than 60°C during drying.