Stable fertilizer for preventing and treating clubroot of rape and preparation method thereof

Abstract

The invention relates to the technical field of agricultural production, in particular to stable fertilizer for preventing and treating clubroot of rape and a preparation method. The stable fertilizer for preventing and treating clubroot of rape comprises the following components in parts by weight: 3-4 parts of a mixed microbial fermentation preparation, 2-4 parts of a biological enzyme condensate, 7-8 parts of borax, 110-180 parts of calcium phosphate, 500-550 parts of urea, 60-80 parts of corn flour, 1-3 parts of montmorillonite and 2-3 parts of polymaleic acid. According to the method, the mixed microbial fermentation preparation and the bio-enzyme solidified substance are combined, and the prevention and treatment effect on the clubroot of the rape is improved on the premise that the physicochemical properties of soil are not damaged.

Description

Technical Field

[0001] The invention relates to the technical field of agricultural production, and in particular to a stable fertilizer for preventing and treating rape clubroot and a preparation method thereof. Background Art

[0002] In recent years, with the rapid development of the economy and the improvement of living standards, more and more people have begun to pay attention to healthy eating issues. Edible oil plays an important role in daily life. It not only provides rich taste and aroma for dishes, but also provides essential fat and energy for the human body. There are many varieties of edible oils, and different types of edible oils have different characteristics and nutritional values. Rapeseed oil is rich in unsaturated fatty acids and vitamin E, and is easily absorbed by the human body. It is a very popular edible oil. However, due to the high risk of rapeseed cultivation, the price of rapeseed oil has been relatively high.

[0003] Rapeseed clubroot is a soil-borne disease caused by infection with Brassica clubroot. This disease has the characteristics of high incidence and wide range of disease, and is the main risk factor in rapeseed cultivation. The pathogen of rapeseed clubroot is a lower organism. The dormant spores of the pathogen are highly resistant to stress and can survive for 6-10 years in the soil, on diseased residues or on the surface of seeds. Soil, diseased residues or seeds containing dormant spores of the pathogen are the primary source of infection in the second year. The dormant spores can be spread with the help of rainwater, irrigation water, agricultural operations, etc., resulting in a serious decline in rapeseed crop yields or even a total crop failure.

[0004] At present, the prevention and control methods of rapeseed clubroot disease mainly include chemical irrigation, crop rotation and soil improvement. Although chemical irrigation is effective quickly, the large amount of chemical agents used causes serious environmental pollution. Although crop rotation is environmentally friendly, it is time-consuming and cannot avoid the re-contamination of the soil by fallen leaves carrying pathogens when they return to the soil. The soil improvement method has a better prevention and control effect, but it is easy to cause soil compaction and a decrease in soil fertility. Summary of the invention

[0005] In view of the technical problem that rapeseed clubroot is difficult to be effectively prevented and controlled without destroying the physical and chemical properties of the soil, the present invention provides a stable fertilizer for preventing and controlling rapeseed clubroot and a preparation method thereof. A mixed microbial fermentation preparation and a bio-enzyme solid are added to the fertilizer, and components containing nutrients such as boron, phosphorus, and nitrogen are used in combination, thereby improving the prevention and control effect of rapeseed clubroot without destroying the physical and chemical properties of the soil.

[0006] In a first aspect, the present invention provides a stable fertilizer for preventing and treating rape clubroot, comprising 3-4 parts by weight of a mixed microbial fermentation preparation, 2-4 parts by weight of a bioenzyme solid, 7-8 parts by weight of borax, 110-180 parts by weight of calcium phosphate, 500-550 parts by weight of urea, 60-80 parts by weight of corn flour, 1-3 parts by weight of montmorillonite and 2-3 parts by weight of polymaleic acid; The mixed microbial fermentation preparation includes a fermentation broth of Streptomyces, a fermentation broth of Streptomyces oxysporus and medical stone powder, wherein the number of viable bacteria in the fermentation broth of Streptomyces is 7×10 8 CFU / g, the number of viable bacteria in the fermentation broth of Streptomyces oxysporum was 5×10 8 CFU / g; The raw materials of the bio-enzyme solidified material include diatomaceous earth, DMPP, bio-enzyme and cross-linking agent. The amount of the cross-linking agent used is sufficient to fix the bio-enzyme. In the bio-enzyme solidified material, the mass fraction of the bio-enzyme is 0.3%-0.8%, and the mass fraction of DMPP is 12.5%-30%.

[0007] Furthermore, the preparation method of the mixed microbial fermentation preparation comprises the following steps: Step 1: mixing the fermentation broth of Streptomyces and the fermentation broth of Streptomyces oxysporum, centrifuging and separating the supernatant; Step 2: Add the supernatant to the medical stone powder and stir evenly to obtain a mixed microbial fermentation preparation.

[0008] Furthermore, the particle size of the medical stone powder is 60-80 meshes.

[0009] Further, the biological enzyme includes chitinase, phosphatase and / or phytase. Chitinase has antifungal activity and can inhibit the mycelial growth and spore germination of the pathogenic bacteria of rape clubroot. Phosphatase can promote the hydrolysis of organic phosphorus compounds in the soil and improve the utilization rate of phosphorus. Phytase can simultaneously improve the utilization rate of phosphorus and calcium. The present invention loads the biological enzyme on diatomaceous earth, which is conducive to improving the stability of the biological enzyme and extending the effective period of the biological enzyme.

[0010] Furthermore, the preparation method of the bio-enzyme solidified material comprises the following steps: S1: using a buffer to dissolve the enzyme to obtain an enzyme buffer; S2: Mix DMPP and diatomaceous earth evenly to obtain a mixed carrier; S2: The bio-enzyme buffer and the mixed carrier are mixed evenly, and then a cross-linking agent is added during the stirring process to carry out a cross-linking reaction. After the cross-linking reaction, the residual liquid is removed, and the bio-enzyme solid is obtained after low-temperature drying.

[0011] Furthermore, the buffer is Tris buffer or PBS buffer, and the pH of the buffer is 5-7; the cross-linking agent is at least one of polyethyleneimine, bis(azobenzidine), epoxy polyamine, polyacrylamide, glutaraldehyde, formaldehyde, succinaldehyde, and acetaldehyde.

[0012] Furthermore, the temperature of the cross-linking reaction is 30-40° C., and the time of the cross-linking reaction is 0.5-1 h.

[0013] In a second aspect, the present invention provides a method for preparing the stable fertilizer as described above, comprising the following steps: Step (1): granulating borax, calcium phosphate, urea and montmorillonite, crushing them, and then mixing them with polymaleic acid to obtain ingredient A; Step (2): uniformly mixing the mixed microbial fermentation preparation, the bio-enzyme solid and the corn flour to obtain ingredient B; Step (3): Add ingredient B to ingredient A and mix well.

[0014] The beneficial effects of the present invention are: The present invention provides a stable fertilizer for preventing and treating rape clubroot and a preparation method thereof, which adopts a method of combining a mixed microbial fermentation preparation and a bioenzyme solidified material, uses bioenzymes to inhibit the mycelial growth and spore germination of the pathogen of rape clubroot, and uses a mixed microbial fermentation preparation to inhibit the growth of the pathogen of rape clubroot, thereby improving the prevention and treatment effect of rape clubroot without destroying the physical and chemical properties of the soil. The present invention adds a nitrification inhibitor 3,4-dipyrazole phosphate (DMPP) to the bioenzyme solidified material, and adds polymaleic acid to the fertilizer, thereby improving the stability of the fertilizer. DETAILED DESCRIPTION

[0015] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0016] The Streptomyces used in the present invention is Streptomyces pyrifos purchased from Yiqiang Biotechnology, with a viable count of 10 billion / g; Streptomyces oxysporum was purchased from Ningbo Mingzhou Biotechnology Co., Ltd., with a product number of BMZ077276. Chitinase was purchased from Hunan Yunbang Biotechnology Co., Ltd., model number YB125065; phosphatase was purchased from Pusrui (Shanghai) Biopharmaceutical Co., Ltd., model number MB12002; phytase was purchased from Shenzhen Hengsheng Biotechnology Co., Ltd., with an enzyme activity of 100,000 u / g.

[0017] Example 1 A stable fertilizer for preventing and treating rape clubroot, the preparation method of which comprises the following steps: 1-1 Preparation of mixed microbial fermentation preparations: Step 1: Mix the fermentation broth of Streptomyces and the fermentation broth of Streptomyces oxysporum and centrifuge at 5000 rpm to separate the supernatant. The number of viable cells in the fermentation broth of Streptomyces is 7×10 8 CFU / g, the number of viable bacteria in the fermentation broth of Streptomyces oxysporum was 5×108 CFU / g.

[0018] Step 2: Add the supernatant to the medical stone powder with a particle size of 60-80 meshes and stir evenly to obtain a mixed microbial fermentation preparation. The mass ratio of the supernatant to the medical stone powder is 1:5.

[0019] 1-2 Preparation of bio-enzyme solidified material: S1: using a Tris buffer having a pH of 5-7 to dissolve chitinase, phosphatase and phytase to obtain a biological enzyme buffer.

[0020] S2: Evenly mix DMPP and diatomaceous earth to obtain a mixed carrier.

[0021] S3: The bio-enzyme buffer and the mixed carrier are mixed evenly, and then glutaraldehyde solution is added during the stirring process for cross-linking reaction, the mass concentration of the glutaraldehyde solution is 4%, the temperature of the cross-linking reaction is 30°C, the time of the cross-linking reaction is 0.5h, and after the cross-linking reaction, the residual liquid is removed by centrifugation, and the bio-enzyme solid is obtained after low-temperature drying. In the bio-enzyme solid, the total mass fraction of chitinase, phosphatase and phytase is 0.3%, and the mass fraction of DMPP is 12.5%.

[0022] 1-3 Preparation of stable fertilizer for preventing and controlling rapeseed clubroot: Step (1): 7 parts by weight of borax, 180 parts by weight of calcium phosphate, 500 parts by weight of urea and 2 parts by weight of montmorillonite are granulated and crushed, and then mixed with 3 parts by weight of polymaleic acid to obtain ingredient A.

[0023] Step (2): 3 parts by weight of a mixed microbial fermentation preparation, 4 parts by weight of a bio-enzyme solid and 60 parts by weight of corn flour are uniformly mixed to obtain ingredient B.

[0024] Step (3): Add ingredient B to ingredient A and mix evenly to obtain fertilizer.

[0025] Example 2 A stable fertilizer for preventing and treating rape clubroot, the preparation method of which comprises the following steps: 2-1 Preparation of mixed microbial fermentation preparations: Step 1: Mix the fermentation broth of Streptomyces and the fermentation broth of Streptomyces oxysporum and centrifuge at 5500 rpm to separate the supernatant. The number of viable cells in the fermentation broth of Streptomyces is 7×10 8 CFU / g, the number of viable bacteria in the fermentation broth of Streptomyces oxysporum was 5×10 8 CFU / g.

[0026] Step 2: Add the supernatant to the medical stone powder with a particle size of 60-80 meshes and stir evenly to obtain a mixed microbial fermentation preparation. The mass ratio of the supernatant to the medical stone powder is 1:3.

[0027] 2-2 Preparation of bio-enzyme solidified material: S1: Dissolve chitinase and phosphatase in PBS buffer with a pH of 5-7 to obtain a biological enzyme buffer.

[0028] S2: Evenly mix DMPP and diatomaceous earth to obtain a mixed carrier.

[0029] S3: The bio-enzyme buffer and the mixed carrier are mixed evenly, and then a 5% epoxy polyamine solution is added during the stirring process for cross-linking reaction. The temperature of the cross-linking reaction is 36°C, and the time of the cross-linking reaction is 0.5h. After the cross-linking reaction, the residual liquid is removed by centrifugation, and the bio-enzyme solid is obtained after low-temperature drying. In the bio-enzyme solid, the total mass fraction of chitinase, phosphatase and phytase is 0.6%, and the mass fraction of DMPP is 20%.

[0030] 2-3 Preparation of stable fertilizer for preventing and controlling rapeseed clubroot: Step (1): 8 parts by weight of borax, 110 parts by weight of calcium phosphate, 550 parts by weight of urea and 1 part by weight of montmorillonite are granulated and then crushed to obtain ingredient A.

[0031] Step (2): 3 parts by weight of a mixed microbial fermentation preparation, 3 parts by weight of a bio-enzyme solid and 80 parts by weight of corn flour are uniformly mixed to obtain ingredient B.

[0032] Step (3): Add ingredient B to ingredient A and mix evenly to obtain fertilizer.

[0033] Example 3 A stable fertilizer for preventing and treating rape clubroot, the preparation method of which comprises the following steps: 3-1 Preparation of mixed microbial fermentation preparations: Step 1: Mix the fermentation broth of Streptomyces and the fermentation broth of Streptomyces oxysporum and centrifuge at 6000 rpm to separate the supernatant. The number of viable cells in the fermentation broth of Streptomyces is 7×10 8 CFU / g, the number of viable bacteria in the fermentation broth of Streptomyces oxysporum was 5×10 8 CFU / g.

[0034] Step 2: Add the supernatant to the medical stone powder with a particle size of 60-80 meshes and stir evenly to obtain a mixed microbial fermentation preparation. The mass ratio of the supernatant to the medical stone powder is 1:7.

[0035] 3-2 Preparation of bio-enzyme solidified material: S1: Dissolve chitinase and phosphatase in PBS buffer with a pH of 5-7 to obtain a biological enzyme buffer.

[0036] S2: Evenly mix DMPP and diatomaceous earth to obtain a mixed carrier.

[0037] S3: The bio-enzyme buffer and the mixed carrier are mixed evenly, and then a 5% epoxy polyamine solution is added during the stirring process for cross-linking reaction. The temperature of the cross-linking reaction is 40°C, and the time of the cross-linking reaction is 1 hour. After the cross-linking reaction, the residual liquid is removed by centrifugation, and the bio-enzyme solid is obtained after low-temperature drying. In the bio-enzyme solid, the total mass fraction of chitinase, phosphatase and phytase is 0.8%, and the mass fraction of DMPP is 25%.

[0038] 3-3 Preparation of stable fertilizer for preventing and controlling rapeseed clubroot: Step (1): 7 parts by weight of borax, 120 parts by weight of calcium phosphate, 540 parts by weight of urea and 3 parts by weight of montmorillonite are granulated and then crushed to obtain ingredient A.

[0039] Step (2): 4 parts by weight of a mixed microbial fermentation preparation, 2 parts by weight of a bio-enzyme solid and 70 parts by weight of corn flour are uniformly mixed to obtain ingredient B.

[0040] Step (3): Add ingredient B to ingredient A and mix evenly to obtain fertilizer.

[0041] Comparative Example 1 A fertilizer for preventing and treating rape clubroot disease, the preparation method of which comprises the following steps: 1-1 Preparation of mixed microbial fermentation preparations: Step 1: Mix the fermentation broth of Streptomyces and the fermentation broth of Streptomyces oxysporum and centrifuge at 5000 rpm to separate the supernatant. The number of viable cells in the fermentation broth of Streptomyces is 7×10 8 CFU / g, the number of viable bacteria in the fermentation broth of Streptomyces oxysporum was 5×10 8 CFU / g.

[0042] Step 2: Add the supernatant to the medical stone powder with a particle size of 60-80 meshes and stir evenly to obtain a mixed microbial fermentation preparation. The mass ratio of the supernatant to the medical stone powder is 1:5.

[0043] 1-2 Preparation of bio-enzyme solidified material: S1: using a Tris buffer having a pH of 5-7 to dissolve chitinase, phosphatase and phytase to obtain a biological enzyme buffer.

[0044] S2: The bio-enzyme buffer and diatomaceous earth are mixed evenly, and then glutaraldehyde solution is added during stirring to perform a cross-linking reaction, wherein the mass concentration of the glutaraldehyde solution is 4%, the temperature of the cross-linking reaction is 30°C, the time of the cross-linking reaction is 0.5h, and after the cross-linking reaction, the residual liquid is removed by centrifugation to obtain a bio-enzyme solid. In the bio-enzyme solid, the total mass fraction of chitinase, phosphatase and phytase is 0.3%.

[0045] 1-3 Preparation of fertilizer for preventing and treating rapeseed clubroot: Step (1): 7 parts by weight of borax, 180 parts by weight of calcium phosphate, 500 parts by weight of urea and 2 parts by weight of montmorillonite are granulated and crushed, and then mixed with 3 parts by weight of polymaleic acid and 0.5 parts by weight of DMPP to obtain ingredient A.

[0046] Step (2): 3 parts by weight of a mixed microbial fermentation preparation, 4 parts by weight of a bio-enzyme solid and 60 parts by weight of corn flour are uniformly mixed to obtain ingredient B.

[0047] Step (3): Add ingredient B to ingredient A and mix evenly to obtain fertilizer.

[0048] Example 4 Treatment group: The stable fertilizers prepared in Examples 1 to 3 and Comparative Example 1 were diluted 80 times with water to obtain diluted fertilizers, and the diluted fertilizers were marked as 1# diluted fertilizer, 2# diluted fertilizer, 3# diluted fertilizer and 4# diluted fertilizer in sequence. The root irrigation method was adopted, and the 1#-4# diluted fertilizers were used to irrigate the roots of rapeseed when transplanting rapeseed. 120 ml of the diluted fertilizer was irrigated to the roots of each rapeseed, and then the roots were irrigated once every 7 days. 100 rapeseeds were irrigated with 1#-4# diluted fertilizers respectively, and the rapeseeds applied with 1# diluted fertilizer, 2# diluted fertilizer, 3# diluted fertilizer and 4# diluted fertilizer were marked as treatment group 1, treatment group 2, treatment group 3 and control group 1, respectively.

[0049] Control group 2: The difference between control group 2 and treatment group 1 was only the application of commercially available compound fertilizer.

[0050] The incidence of rapeseed clubroot and soil agglomeration in treatment group 1-treatment group 3 and control group 1-control group 2 were statistically analyzed, and the incidence of rapeseed clubroot in treatment group 1-treatment group 3 and control group 1-control group 2 was calculated. The statistical results are shown in Table 1.

[0051] Table 1 The incidence of rapeseed clubroot in treatment groups 1-3 and control groups 1-2

[0052] Test Example 1 Example 1 and Comparative Example 1 were stored at room temperature, and the residual rate of DMPP in the fertilizer was measured every 7 days. The measurement results are shown in Table 2.

[0053] Table 2 Residual rate of DMPP in the fertilizers of Example 1 and Comparative Example 1

[0054] Although the present invention has been described in detail by way of preferred embodiments, the present invention is not limited thereto. Without departing from the spirit and essence of the present invention, a person of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and such modifications or substitutions shall be within the scope of the present invention. Any person of ordinary skill in the art may easily conceive of changes or substitutions within the technical scope disclosed by the present invention, and such changes or substitutions shall be within the scope of protection of the present invention.

Claims

1. A stable fertilizer for preventing and treating rape clubroot, characterized in that: The invention comprises 3-4 parts by weight of a mixed microbial fermentation preparation, 2-4 parts by weight of a bioenzyme solid, 7-8 parts by weight of borax, 110-180 parts by weight of calcium phosphate, 500-550 parts by weight of urea, 60-80 parts by weight of corn flour, 1-3 parts by weight of montmorillonite and 2-3 parts by weight of polymaleic acid; The mixed microbial fermentation preparation includes a fermentation broth of Streptomyces, a fermentation broth of Streptomyces oxysporus and medical stone powder, wherein the number of viable bacteria in the fermentation broth of Streptomyces is 7×10 8 CFU / g, the number of viable bacteria in the fermentation broth of Streptomyces oxysporum was 5×10 8 CFU / g; The raw materials of the bio-enzyme solidified material include diatomaceous earth, DMPP, bio-enzyme and cross-linking agent. In the bio-enzyme solidified material, the mass fraction of the bio-enzyme is 0.3%-0.8%, and the mass fraction of DMPP is 12.5%-30%.

2. The stable fertilizer for preventing and treating rape clubroot as claimed in claim 1, characterized in that: The preparation method of the mixed microbial fermentation preparation comprises the following steps: Step 1: mixing the fermentation broth of Streptomyces and the fermentation broth of Streptomyces oxysporum, centrifuging and separating the supernatant; Step 2: Add the supernatant to the medical stone powder and stir evenly to obtain a mixed microbial fermentation preparation.

3. The stable fertilizer for preventing and treating rape clubroot as claimed in claim 1, characterized in that: The particle size of medical stone powder is 60-80 mesh.

4. The stable fertilizer for preventing and treating rape clubroot as claimed in claim 1, characterized in that: The biological enzyme includes chitinase, phosphatase and / or phytase.

5. The stable fertilizer for preventing and treating rape clubroot as claimed in claim 1, characterized in that: The preparation method of the bio-enzyme solidified material comprises the following steps: S1: using a buffer to dissolve the enzyme to obtain an enzyme buffer; S2: Mix DMPP and diatomaceous earth evenly to obtain a mixed carrier; S3: Evenly mix the bio-enzyme buffer and the mixed carrier, then add a cross-linking agent during the stirring process to carry out a cross-linking reaction, remove the residual liquid after the cross-linking reaction, and obtain the bio-enzyme solid after low-temperature drying.

6. The stable fertilizer for preventing and treating rape clubroot as claimed in claim 5, characterized in that: The buffer is Tris buffer or PBS buffer; the cross-linking agent is at least one of polyethyleneimine, bis(azobenzidine), epoxy polyamine, polyacrylamide, glutaraldehyde, formaldehyde, succinaldehyde and acetaldehyde.

7. The stable fertilizer for preventing and treating rape clubroot as claimed in claim 5, characterized in that: The temperature of the cross-linking reaction is 30-40°C, and the time of the cross-linking reaction is 0.5-1h.

8. A method for preparing a stable fertilizer according to any one of claims 1 to 7, characterized in that: The steps include: Step (1): granulating borax, calcium phosphate, urea and montmorillonite, crushing them, and then mixing them with polymaleic acid to obtain ingredient A; Step (2): uniformly mixing the mixed microbial fermentation preparation, the bio-enzyme solid and the corn flour to obtain ingredient B; Step (3): Add ingredient B to ingredient A and mix well.

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