Agricultural microbial agent and its preparation method and application

Abstract

The present invention belongs to the technical field of microbial fertilizers, and in particular, relates to an agricultural microbial agent and a preparation method and application thereof, wherein the agricultural microbial agent comprises the following raw materials in mass percentage: 4-8% brown sugar, 1-3% glucose, 12-18% peptone, 8-12% beef extract, 0.5-1.5% edible salt, 8-12% yeast extract, 1-3% Bacillus megaterium, 1-3% peptone-like bacillus, 1-3% Bacillus subtilis, and the balance is water. The number of effective live bacteria in the agricultural microbial agent of the present invention is ≥200 million / mL, and the rate of foreign bacteria is ≤10%, which meets the use standard of the agricultural microbial agent. The application of the microbial agent of the present invention can not only significantly increase the yield of peppers and increase economic benefits, but also significantly increase the length, width and flesh thickness of peppers, reduce the nitrate content in peppers, and make its content far lower than the maximum limit standard of nitrate in pollution-free peppers, effectively improve the quality of peppers, and effectively reduce the amount of chemical fertilizers, and has various functions.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial fertilizers, and in particular relates to an agricultural microbial agent and a preparation method and application thereof. Background Art

[0002] my country is a large agricultural producer. In the agricultural production process, fertilizers are needed to supplement the necessary elements for crop production. However, in pursuit of high yields and increased income, humans apply a large amount of traditional fertilizers to the soil, which not only damages the soil to a certain extent, but also has poor absorption and utilization of traditional fertilizers by crops. In Hainan, continuous pepper cropping and high-water and high-fertilizer management have caused soil acidification and soil microbial flora imbalance, which seriously affect the sustainable production of Hainan peppers.

[0003] Microbial fertilizer is a product that uses the life activities of microorganisms to cause crops to obtain specific fertilizer effects. It is a type of fertilizer used in agricultural production. Microbial fertilizer contains a large number of beneficial microorganisms, which can improve crop nutritional conditions, fix nitrogen and activate some ineffective nutrients in the soil, creating a good soil microecological environment to promote crop growth.

[0004] However, most of the microbial agents on the market currently have relatively simple functions, which are mainly to improve soil and increase yield, but cannot improve the quality of crops or significantly reduce the amount of fertilizers used. Therefore, the present invention provides an agricultural microbial agent that can effectively solve the above defects. Summary of the invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and to provide an agricultural microbial agent and a preparation method and application thereof. The agricultural microbial agent of the present invention has various functions. The application of the microbial agent of the present invention can not only significantly increase the yield of peppers and increase economic benefits, but also significantly increase the length, width and flesh thickness of peppers, reduce the nitrate content in peppers, make its content far lower than the maximum limit standard of nitrate in pollution-free peppers, effectively improve the quality of peppers, and effectively reduce the amount of fertilizer used.

[0006] The object of the present invention is achieved in this way: an agricultural microbial agent comprises the following raw materials in percentage by mass: 4-8% of brown sugar, 1-3% of glucose, 12-18% of peptone, 8-12% of beef extract, 0.5-1.5% of edible salt, 8-12% of yeast extract, 1-3% of Bacillus megaterium, 1-3% of Paenibacillus peptoneus, 1-3% of Bacillus subtilis, and the balance is water.

[0007] Preferably, the agricultural microbial agent comprises the following raw materials in percentage by mass: 5-7% brown sugar, 1.5-2.5% glucose, 14-16% peptone, 9-11% beef extract, 0.8-1.2% edible salt, 9-11% yeast extract, 1.5-2.5% Bacillus megaterium, 1.5-2.5% Paenibacillus peptoneus, 1.5-2.5% Bacillus subtilis, and the balance is water.

[0008] Preferably, the agricultural microbial agent comprises the following raw materials in percentage by mass: 6% brown sugar, 2% glucose, 15% peptone, 10% beef extract, 1% edible salt, 10% yeast extract, 2% Bacillus megaterium, 2% Paenibacillus peptoneus, 2% Bacillus subtilis, and 50% water.

[0009] Preferably, the number of effective live bacteria in the agricultural microbial agent is ≥ 200 million / mL, and the rate of foreign bacteria is ≤ 10%.

[0010] The present invention also discloses a method for preparing the agricultural microbial agent, which comprises the following steps:

[0011] (1) Sterilize all equipment used;

[0012] (2) preparing and sterilizing the first-level seed tank, and transferring the seed culture medium into the first-level seed tank for culturing;

[0013] (3) Transfer all materials in the primary seed tank to the sterilized secondary seed tank, inoculate the shake flask strain into the secondary seed tank for cultivation and primary fermentation;

[0014] (4) transferring all materials in the secondary seed tank to the fermentation tank for secondary fermentation;

[0015] (5) The fermented bacterial liquid is sampled and tested after fermentation. After all indicators are tested to be qualified, it is put into a storage tank, filled and packaged into a finished product, and an agricultural microbial agent is obtained.

[0016] Preferably, the agricultural microbial agent is in the form of an aqueous solution.

[0017] Preferably, the agricultural microbial agent is used in pepper cultivation by flushing, and the agricultural microbial agent is diluted into a 200-300 times liquid.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] 1. The number of effective live bacteria in the agricultural microbial agent of the present invention is ≥ 200 million / mL, and the rate of foreign bacteria is ≤ 10%, which meets the use standards of agricultural microbial agents.

[0020] 2. Field tests have shown that the application of the agricultural microbial agent of the present invention can significantly increase the yield of peppers, with an increase of 20.1-20.6%; the application of the agricultural microbial agent of the present invention can also significantly increase the fruit length, fruit width and flesh thickness of peppers. Compared with conventional fertilization, the fruit length increases by 6.77-7.81%, the fruit width increases by 9.64-9.59%; the flesh thickness increases by 16.2-21.1%; based on the purchase price of 3.5 yuan / kg in 2018 and 4 yuan / kg in 2019, excluding production costs, the application of the agricultural microbial agent of the present invention can increase the income per mu of peppers by 710.6-891.8 yuan compared with conventional fertilization, which is 32.8-36.1% higher than conventional fertilization.

[0021] 3. The application of the agricultural microbial agent of the present invention can effectively reduce the amount of chemical fertilizers used, and the application of the agricultural microbial agent of the present invention can significantly reduce the nitrate content in peppers, making its content far lower than the maximum limit of nitrate in pollution-free peppers, and can improve the quality of crops. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The present invention is a flow chart for preparing the agricultural microbial agent. DETAILED DESCRIPTION

[0023] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0024] The raw materials used in the embodiments of the present invention are as follows, but are not limited to the following raw materials. The present invention only uses the following raw materials as specific examples to further illustrate the effects of the agricultural microbial agent of the present invention:

[0025] Brown sugar, glucose, edible salt: local;

[0026] Peptone, beef extract, yeast extract: Shandong Yubao Biotechnology Co., Ltd.;

[0027] Bacillus megaterium, Bacillus peptoneus, Bacillus subtilis: self-produced.

[0028] The main equipment used in the present invention is shown in the following table:

[0029] Name of main production equipment Brand Model quantity Name of production unit First level seed tank 20L 1 Yixin Pharmaceutical Secondary seed tank 200L 1 Yixin Pharmaceutical Fermentation tank 2000L 1 Yixin Pharmaceutical Storage tank 3000L 2 Yixin Pharmaceutical Electric steam boiler 108KW 2 Shanghai Huazheng gas tank 1M3 / 8kg 1 Shanghai Shenjiang Screw air compressor TH22A-7(22kw) 1 Shanghai Bright

[0030] The present invention sets embodiments 1 to 5, and the feeding ratio is shown in the following table:

[0031]

[0032] The preparation methods of Examples 1 to 5 include the following steps:

[0033] (1) Sterilize all equipment used;

[0034] (2) preparing and sterilizing the first-level seed tank, and transferring the seed culture medium into the first-level seed tank for culturing;

[0035] (3) Transfer all materials in the primary seed tank to the sterilized secondary seed tank, inoculate the shake flask strain into the secondary seed tank for cultivation and primary fermentation;

[0036] (4) transferring all materials in the secondary seed tank to the fermentation tank for secondary fermentation;

[0037] (5) The fermented bacterial liquid is sampled and tested after fermentation. After all indicators are tested to be qualified, it is put into a storage tank, filled and packaged into a finished product, and an agricultural microbial agent is obtained.

[0038] After testing, the number of effective live bacteria in the agricultural microbial agents prepared in Examples 1 to 5 of the present invention is ≥ 200 million / mL, and the rate of foreign bacteria is ≤ 10%, which meets the use standards of agricultural microbial agents.

[0039] Test example

[0040] The effect of the product of the embodiment is verified by taking the application of the product of the embodiment on peppers as an example.

[0041] The agricultural microbial agent of the embodiment is used on peppers to test the fertilizer effect and safety, and the fertilizer effect and safety of the product are evaluated from the aspects of yield and economic benefit, so as to provide a scientific basis for the registration, promotion and application of the agricultural microbial agent.

[0042] Experimental location: Xi'an Village, Qiaotou Town, Chengmai County, Hainan Province.

[0043] Trial period: November 2017-April 2018, November 2018-April 2019.

[0044] Soil analysis: The soil in the test field is paddy soil developed from coastal sediments, and the soil fertility is medium. Before the test, the soil of the tillage layer (0-20cm) was taken for soil analysis of various indicators: organic matter was determined by potassium dichromate volumetric method-external heating method; pH value analysis was determined by water-soil ratio of 2.5:1 and composite electrode; alkaline hydrolysis nitrogen was determined by alkaline diffusion method; available phosphorus was extracted with hydrochloric acid-ammonium fluoride and determined by molybdenum antimony colorimetry; available potassium was extracted with ammonium acetate and determined by flame photometry. The results of soil physical and chemical properties analysis were: soil organic matter 17.5g / kg, pH 5.30, alkaline hydrolysis nitrogen 154.0mg / kg, available phosphorus 80.1mg / kg, available potassium 68.8mg / kg.

[0045] Test crop: Guyu pepper 109.

[0046] Fertilizers tested:

[0047] Commercial organic fertilizer, compound fertilizer (base fertilizer is 15-15-15, topdressing is 16-6-18), superphosphate (containing P2O5 16%).

[0048] Agricultural microbial agent, produced and provided by Hainan Wanhebao Biotechnology Co., Ltd., has an effective live bacteria count of ≧200 million / ml and is in liquid form.

[0049] Experimental design and fertilization method:

[0050] Experimental design: A small plot experiment was adopted, with a length of 6.67m, a width of 3.0m and an area of ​​20m 2 The peppers in the plot are planted in furrows and ridges. Peppers are planted on the ridges, and the furrows between the ridges are used for topdressing and watering. Each plot is built with 2 ridges and 3 furrows. The ridge width is 90cm and the furrow width is 40cm. 1.5m wide protective rows are set around the experimental site.

[0051] The experiment includes 4 treatments, which are as follows:

[0052] Treatment 1: blank control, no fertilizer (CK);

[0053] Treatment 2: local conventional fertilization (F1);

[0054] Treatment 3: Conventional fertilizer reduced by 20% + test fertilizer (F2)

[0055] Treatment 4: Conventional fertilizer reduction of 20% + agricultural microbial inoculants to inactivate the matrix (F3)

[0056] The specific application method and management measures of Treatment 4 are the same as those of Treatment 3.

[0057] Three replicate plots were set up for each treatment, and the plots were arranged in random blocks. Pepper seedlings were raised on October 1, 2017 and October 3, 2018, and transplanted on November 15, 2017 and November 17, 2018. Pepper seedlings were planted on ridges, with two rows of peppers transplanted per ridge, with a plant spacing of 0.40 m and a row spacing of 0.70 m.

[0058] Fertilization method:

[0059] Base fertilizer: 40kg 15-15-15 triple compound fertilizer, 50kg superphosphate and 250kg commercial organic fertilizer per mu.

[0060] Topdressing: Topdressing was performed every 15 days after transplantation, with the amount of topdressing being 18 kg / mu each time. Treatment 2 used 16-6-18 compound fertilizer for conventional fertilization, treatment 3 used 16-6-18 compound fertilizer mixed with the microbial agent of Example 3, wherein the microbial agent was diluted to 250 times liquid, and treatment 4 used 16-6-18 compound fertilizer mixed with the agricultural microbial agent inactivated matrix of Example 3. When topdressing, the fertilizer was spread in the ditch and then watered.

[0061] Measurement and statistical analysis methods:

[0062] (1) Yield survey: The first harvest and yield were calculated on January 20, 2018 and January 23, 2019, and the subsequent harvest and yield were calculated on February 3, February 17, March 2, and March 16, 2018, and February 6, February 20, March 3, and March 18, 2019. The pepper yield in the plot was converted into per mu yield. The blank control was a control without any fertilizer application, and the yield was based on the basic soil yield.

[0063] (2) Analyze the nitrate content of pepper by ultraviolet spectrophotometry.

[0064] (3) Statistical analysis method: SPSS V17.0 statistical analysis software was used for analysis.

[0065] Test results:

[0066] 1. Effects of different fertilization treatments on pepper yield

[0067] The average yield of peppers treated with agricultural microbial agents in 2018 and 2019 was 1567.1kg / mu and 1616.4kg / mu, an increase of 609.3kg / mu and 605.0kg / mu compared with the control, with an increase rate of 63.6% and 59.8%, respectively, and an increase of 267.6kg / mu and 270.7kg / mu compared with conventional fertilization, with an increase rate of 20.6% and 20.1% (Table 1). Variance analysis showed that the yield-increasing effect reached a significant level. The yield of peppers treated with water spraying was equivalent to that of conventional treatment, indicating that agricultural microbial agents can significantly increase pepper yields.

[0068] Table 1 Effects of different treatments on pepper yield

[0069]

[0070]

[0071] 2. Effects of different fertilization treatments on economic traits of pepper

[0072] Application of agricultural microbial agents to peppers can significantly increase fruit length, fruit width and flesh thickness. Compared with conventional fertilization, the indicators increased by 1.30 cm, 0.35 cm and 0.06 cm in 2018, with growth rates of 6.77%, 9.64% and 16.2%, respectively; in 2019, the indicators increased by 1.50 cm, 0.35 cm and 0.08 cm, with increase rates of 7.81%, 9.59% and 21.1%, respectively (Table 2). Effects of sterilized substrate treatment on pepper fruit traits Compared with conventional fertilization, fruit length increased by 4.69%, fruit width increased by 4.68%, and flesh thickness increased by 5.41% in 2018; fruit length increased by 5.73%, fruit width increased by 5.21%, and flesh thickness increased by 5.26% in 2019. At the same time, the application of agricultural microbial agents can also significantly reduce the nitrate content in peppers, with a decrease of 37.6% and 39.7% in 2018 and 2019, respectively (Table 2). In summary, the application of agricultural microbial agents to peppers can significantly improve the economic traits of peppers and reduce the nitrate content in fruits.

[0073] Table 2 Effects of different treatments on pepper fruit traits

[0074]

[0075] 3. Economic benefits of applying agricultural microbial agents

[0076] Combined with the input cost, the economic benefits of each treatment were analyzed (Table 3). The application of agricultural microbial agents to peppers can significantly improve economic benefits, with an average increase of 2,680.9 yuan and 3,609.6 yuan per mu in 2018 and 2019, respectively, which is 36.1% and 32.8% higher than the 1,970.3 yuan and 2,717.8 yuan of conventional fertilization treatments. This shows that the application of agricultural microbial agents to peppers can bring good economic benefits.

[0077] Table 3 Comparison of economic benefits of peppers treated with different fertilizers

[0078]

[0079] Note: The average price of pepper in 2018 and 2019 was 3.5 yuan / kg and 4 yuan / kg

[0080] In summary, field tests show that the application of the agricultural microbial agent of the present invention can significantly increase the yield of peppers, with an increase of 20.1-20.6%. At the same time, the application of the agricultural microbial agent of the present invention also significantly increases the length, width and flesh thickness of peppers. Compared with conventional fertilization, the fruit length increases by 6.77-7.81%, the fruit width increases by 9.64-9.59%; the flesh thickness increases by 16.2-21.1%. Finally, the application of the agricultural microbial agent of the present invention significantly reduces the nitrate content in peppers, making its content far lower than the maximum limit of nitrate in pollution-free peppers. Based on the purchase price of 3.5 yuan / kg in 2018 and 4 yuan / kg in 2019, excluding the production cost, the application of the agricultural microbial agent of the present invention increases the income per mu of peppers by 710.6-891.8 yuan compared with conventional fertilization, which is 32.8-36.1% higher than conventional fertilization. Overall, the application of the agricultural microbial agent of the present invention to peppers can significantly increase yield, improve economic properties, reduce nitrate content, and increase economic benefits compared to conventional fertilization. Therefore, the application of the agricultural microbial agent produced by the present invention can significantly improve the economic properties of peppers, increase yield, and increase economic benefits.

Claims

1. An agricultural microbial agent, characterized in that: The invention comprises the following raw materials in percentage by mass: 4-8% brown sugar, 1-3% glucose, 12-18% peptone, 8-12% beef extract, 0.5-1.5% edible salt, 8-12% yeast extract, 1-3% Bacillus megaterium, 1-3% Paenibacillus peptoneus, 1-3% Bacillus subtilis, and the balance is water.

2. The agricultural microbial agent according to claim 1, characterized in that: The invention comprises the following raw materials in percentage by mass: 5-7% brown sugar, 1.5-2.5% glucose, 14-16% peptone, 9-11% beef extract, 0.8-1.2% edible salt, 9-11% yeast extract, 1.5-2.5% Bacillus megaterium, 1.5-2.5% Paenibacillus peptoneus, 1.5-2.5% Bacillus subtilis, and the balance is water.

3. The agricultural microbial agent according to claim 1, characterized in that: The invention comprises the following raw materials in percentage by weight: 6% of brown sugar, 2% of glucose, 15% of peptone, 10% of beef extract, 1% of edible salt, 10% of yeast extract, 2% of Bacillus megaterium, 2% of Paenibacillus peptoneus, 2% of Bacillus subtilis and 50% of water.

4. The agricultural microbial agent according to any one of claims 1 to 3, characterized in that The number of effective live bacteria in the agricultural microbial agent is ≥ 200 million / mL, and the rate of foreign bacteria is ≤ 10%.

5. A method for preparing the agricultural microbial agent as claimed in claim 4, characterized in that: The following steps are involved: (1) Sterilize all equipment used; (2) preparing and sterilizing the first-level seed tank, and transferring the seed culture medium into the first-level seed tank for culturing; (3) Transfer all materials in the primary seed tank to the sterilized secondary seed tank, inoculate the shake flask strain into the secondary seed tank for cultivation and primary fermentation; (4) transferring all materials in the secondary seed tank to the fermentation tank for secondary fermentation; (5) The fermented bacterial liquid is sampled and tested after fermentation. After all indicators are tested to be qualified, it is put into a storage tank, filled and packaged into a finished product, and an agricultural microbial agent is obtained.

6. The agricultural microbial agent according to claim 4, characterized in that: The agricultural microbial agent is in the form of an aqueous solution.

7. Use of the agricultural microbial agent as claimed in claim 6 in pepper cultivation.

8. The use of the agricultural microbial agent in pepper planting as claimed in claim 7, characterized in that: The agricultural microbial agent is applied by flushing.

9. The use of the agricultural microbial agent in pepper planting as claimed in claim 8, characterized in that: The agricultural microbial agent is diluted to 200 to 300 times liquid.

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