Compound microbial fertilizer and application thereof in field planting

Abstract

The invention discloses a compound microbial fertilizer and application thereof in field planting. The survival rate of the paenibacillus mucilaginosus HX-02 disclosed by the invention can be up to 62.8% after 170d under the condition of 38% of nitrogen, phosphorus and potassium. The strain is organically combined with bacillus velezensis RKN1111 and a nitrogen-phosphorus-potassium inorganic fertilizer (N + P2O5 + K2O = 38%), the compound microbial fertilizer is developed, the viable count of effective bacteria is not less than 1.5 billion / g, and the viable count of paenibacillus mucilaginosus is not less than 0.5 billion / g. The compound microbial fertilizer taking the strain as a main functional bacterium is applied to field corn planting, can replace part of chemical fertilizers and increase soil fertility, can reduce the use amount of the chemical fertilizers by 15.56%, and achieves the effects of promoting the growth of field crops and increasing the yield by 10.2%. The paenibacillus mucilaginosus HX-02 is separated from soil, is harmless to human, livestock and environment, and meets the requirements of green plant protection and agricultural sustainable development.

Description

Technical Field

[0001] The invention belongs to the technical field of agriculture, and in particular relates to a composite microbial fertilizer and application thereof in field planting. Background Art

[0002] In the process of land cultivation, large-scale and excessive application of chemical fertilizers, arbitrary use of inferior manure, and long-term flooding have led to soil acidification, salinization, and compaction, and the soil's ability to provide fertilizer has decreased. In recent years, the use of microbial compound agents has become a more ideal solution. Microbial compound agents generally have the functions of improving soil structure, transforming soil nutrients, reducing crop diseases and pests, promoting plant growth, and increasing crop yields, and are increasingly valued by growers. Long-term use of microbial compound agents can not only improve soil aggregate structure, regulate soil pH, loosen the soil to promote plant root development, but also enhance plant absorption of water and nutrients, reduce the use of chemical fertilizers and pesticides, and achieve green and sustainable agricultural development. Summary of the invention

[0003] The purpose of the invention is to provide a composite microbial fertilizer and application thereof in field planting.

[0004] In order to achieve the purpose of the present invention, in the first aspect, the present invention provides a composite microbial agent, the active ingredient of which is jelly-like bacillus ( Paenibacillus mucilaginosus ) HX-02 and Bacillus velezinoffe ( Bacillus velezensis )RKN1111.

[0005] The jelly-like bacillus of the present invention ( Paenibacillus mucilaginosus )HX-02, now deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postal Code 100101, Deposit Number CGMCC No.30949, Deposit Date June 14, 2024.

[0006] Jelly-like Bacillus HX-02 can be used in an environment with 38% nitrogen, phosphorus and potassium.

[0007] Bacillus jelly HX-02 can replace some fertilizers, increase soil fertility, promote nutrient absorption during the growth stage of field crops, and increase yields.

[0008] Bacillus Velez Bacillus velezensis ) The deposit number of RKN1111 is CGMCC No. 29300. Bacillus velezensis RKN1111 has been disclosed in CN118389321A.

[0009] In a second aspect, the present invention provides a biological fertilizer or a plant growth promoter prepared from the composite microbial agent.

[0010] In a third aspect, the present invention provides a composite microbial fertilizer, which is composed of the composite microbial agent, nitrogen, phosphorus and potassium inorganic fertilizers and organic matter.

[0011] The number of live bacteria of Bacillus jelly-like in the composite microbial fertilizer is 30 million to 70 million / g (preferably 50 million / g), and the number of live bacteria of Bacillus velez is 50 million to 150 million / g (preferably 100 million / g).

[0012] The organic matter can be selected from one or more of humic acid, bentonite, peat and the like.

[0013] Furthermore, the nitrogen, phosphorus and potassium inorganic fertilizer is a chemical fertilizer with N+P2O5+K2O=38%.

[0014] Furthermore, the nitrogen, phosphorus and potassium inorganic fertilizer is a mixture of N, P2O5 and K2O in a mass ratio of 20:10:8.

[0015] The preparation method of the composite microbial fertilizer comprises: fermenting and culturing the jelly-like Paenibacillus HX-02 and the Velez Bacillus RKN1111 respectively, centrifugally concentrating and drying the fermented liquid to prepare jelly-like Paenibacillus powder and Velez Bacillus powder respectively, then mixing the two powders at a mass ratio of 5:1, and compounding the obtained composite powder with nitrogen, phosphorus and potassium inorganic fertilizers and organic matter; Among them, the mass ratio of composite bacterial powder, nitrogen, phosphorus and potassium inorganic fertilizers and organic matter is 1: (120-145): (20-40), preferably 1: 138: 28.

[0016] Furthermore, the formula of the culture medium for fermenting and culturing the jelly-like Bacillus is: 15 g starch, 2.0 g (NH4)2SO4, 2.0 g KH2PO4, 1.5 g yeast extract, 0.5 g FeCl3, 1000 mL water, pH 7.5.

[0017] Furthermore, the formula of the culture medium for fermenting and culturing the Bacillus Velez subtilis is: 10.0 g starch, 20.0 g bean cake powder, 4.0 g MgSO4, 2.0 g yeast extract, 1000 mL water, pH 7.0.

[0018] In a fourth aspect, the present invention provides application of the composite microbial fertilizer in field planting.

[0019] In a fifth aspect, the present invention provides the use of the composite microbial fertilizer in increasing soil fertility and promoting plant growth.

[0020] Such plants include, but are not limited to, corn and wheat.

[0021] By means of the above technical solution, the present invention has at least the following advantages and beneficial effects: The jelly-like Paenibacillus HX-02 of the present invention can survive for 170 days under the condition of 38% nitrogen, phosphorus and potassium, with a survival rate of up to 62.8%. When applied to the composite microbial fertilizer (composite microbial agent + fertilizer + organic matter) in field corn and wheat planting, the effective live bacteria of the jelly-like Paenibacillus are not less than 50 million / g, which can reduce the amount of fertilizer by 15.56%. The strain HX-02 can replace part of the fertilizer and increase soil fertility, achieving the effect of promoting corn production by 10.2% and wheat production by 12.4%. The jelly-like Paenibacillus HX-02 is separated from the soil, is harmless to humans, animals and the environment, and meets the needs of green plant protection and sustainable agricultural development. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The figure shows the change of effective viable bacteria count of Paenibacillus gelatinosa in mixtures of different fertilizer concentrations in a preferred embodiment of the present invention. DETAILED DESCRIPTION

[0023] The present invention aims to provide a composite microbial fertilizer, jelly-like Paenibacillus HX-02, which is used in combination with chemical fertilizers and its application in field planting.

[0024] The present invention provides a strain HX-02 isolated from soil, which is classified and named as Paenibacillus jelly-like ( Paenibacillus mucilaginosus ), has been deposited in the General Microbiology Center of China Microorganism Culture Collection Committee, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postal Code 100101, with deposit number CGMCC No.30949 and deposit date June 14, 2024.

[0025] The present invention also provides a biological fertilizer or a plant growth promoter prepared from the jelly-like Paenibacillus or a bacterial agent containing the jelly-like Paenibacillus.

[0026] The invention also provides an application of the jelly-like Paenibacillus, in which the survival rate is as high as 62.8% under the condition of 38% nitrogen, phosphorus and potassium.

[0027] The invention also provides application of the jelly-like Paenibacillus composite microbial fertilizer in field planting.

[0028] The jelly-like Paenibacillus HX-02 of the invention has the functions of high salt tolerance, reducing the amount of chemical fertilizers, promoting plant growth and increasing yield.

[0029] The jelly-like Paenibacillus HX-02 of the present invention is compounded with chemical fertilizer to form a composite microbial fertilizer, which is conducive to colonization in the soil. In addition, the composite microbial fertilizer provided by the present invention has a reasonable formula design and is easy to produce and manufacture.

[0030] Strain HX-02 is isolated from the soil and is harmless to humans, animals and the environment, meeting the needs of green plant protection and sustainable agricultural development.

[0031] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0032] Example 1 Storage period of mixtures of jelly-like Paenibacillus sp. HX-02 and different fertilizers

[0033] 1. Experimental methods

[0034] Treatment group settings: Equal amounts of (NH4)2SO4, KH2PO4 and K2SO4 (the mass ratio of (NH4)2SO4, KH2PO4 and K2SO4 is 1:1:1) are mixed evenly, and then a certain amount of humic acid is added to make 500g of a mixture with different [(NH4)2SO4+KH2PO4+ K2SO4] concentrations. Finally, 22.5g of jelly-like Bacillus powder is added, and the effective live bacteria count is 10 billion / g.

[0035] The preparation method of jelly-like Paenibacillus powder is as follows: (1) Preparation of jelly-like Paenibacillus HX-02 fermentation broth

[0036] The jelly-like fermentation broth of Paenibacillus sp. HX-02 was prepared by secondary submerged fermentation.

[0037] a. Seed tank culture

[0038] Culture medium: starch 5.0 g, (NH4)2SO4 0.5 g, KH2PO4 2.0 g, yeast extract 1.5 g, FeCl3 0.5 g, water 1000 mL, pH 7.5.

[0039] Culture conditions: tank pressure 0.5MPa, culture temperature 28℃, ventilation ratio 1:0.3.

[0040] b. Fermentation tank culture

[0041] Culture medium: starch 15 g, (NH4)2SO4 2.0 g, KH2PO4 2.0 g, yeast extract 1.5 g, FeCl3 0.5 g, water 1000 mL, pH 7.5; Culture conditions: tank pressure 0.5MPa, culture temperature 30℃, ventilation ratio 1:0.5-1.0.

[0042] (1) Preparation of jelly-like Bacillus HX-02 bacterial powder

[0043] The jelly-like Bacillus HX-02 strain fermentation broth prepared above was subjected to a post-treatment process combining centrifugal concentration and rotary flash drying to prepare a high-bacteria-content bacterial powder with an effective live bacterial count of 10 billion / g.

[0044] A compound microbial fertilizer was developed. This experiment set up 5 treatments, namely T1, T2, T3, T4 and T5, each treatment was repeated 3 times, and stored at room temperature. The concentrations of T1, T2, T3, T4 and T5 are as follows: T1: [(NH4)2SO4+KH2PO4+ K2SO4]=0

[0045] T2: [(NH4)2SO4+KH2PO4+ K2SO4]=8%

[0046] T3: [(NH4)2SO4+KH2PO4+ K2SO4]=18%

[0047] T4: [(NH4)2SO4+KH2PO4+ K2SO4]=28%

[0048] T5: [(NH4)2SO4+KH2PO4+ K2SO4]=38%

[0049] Jelly-like Paenibacillus detection: Take out the sample, mix it thoroughly, weigh 10.0g of the sample accurately with a sterile medicine spoon, add it to 100.0mL of sterile water, let it stand for 20min, then place it on a shaker, shake it at 200r / min for 30min to make a bacterial suspension. Use a 5mL sterile pipette to add it to 45mL of sterile water, and continue to dilute it in a gradient of 1:10 to obtain 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 The bacterial suspension was diluted to 10 -6 and 10 -7 Dilution, use a 0.5 mL sterile pipette to take 0.1 mL of bacterial suspension, and evenly spread it on the test medium plate of jelly-like Bacillus. Each dilution is repeated 3 times, and sterile water is used as a blank control. Incubate at 32℃ for 48h, count the number of colonies on the plate, and calculate the effective bacterial content of jelly-like Bacillus based on the average value.

[0050] 2. Experimental results

[0051] The mixtures containing jelly-like Paenibacillus with different fertilizer ratios prepared by the above method were stored at room temperature, and samples were taken at 0, 22d, 56d, 78d, 120d and 170d, respectively, to determine the effective bacterial content. The preservation effect of jelly-like Paenibacillus in mixtures with different fertilizer concentrations is shown in Table 1, and the change of the effective live bacterial count of jelly-like Paenibacillus in mixtures with different fertilizer concentrations is shown in Table Figure 1 .

[0052] Table 1 Preservation effect of jelly-like Paenibacillus in mixtures of different fertilizer concentrations

[0053] Example 2 Screening of highly efficient compound strains of strain HX-02

[0054] The experiment was conducted in Dongxu Village, Boye County, at 115°30″ east longitude and 38°30″ north latitude. The soil is light loamy tidal brown soil and the area is 3082.5m 2 The corn variety was Zhengdan 958. The experiment was conducted with 5 treatments and 3 replications.

[0055] The treatment groups were set up as follows: T1: 50 kg of conventional corn compound fertilizer (N+P2O5+K2O=45%), purchased from Baoding Watson Fertilizer Manufacturing Co., Ltd.

[0056] T2: FHF + HX-02 + JS; T3: FHF+HX-02+RKN1111; T4: FHF+HX-02; T5: FHF+HX-02+RKN1111+JS.

[0057] FHF is a compound fertilizer (N+P2O5+K2O=38%), HX-02, RKN1111, JS (Bacillus licheniformis, see Ren Yue, Cai Binhai, Zhang Aimin. Antagonistic effect of thiamethoxam on jelly-like Bacillus, Bacillus subtilis and Bacillus licheniformis [J]. Anhui Agricultural Sciences, 2019, 47(05): 1-5. Bacillus licheniformis JS was kindly donated by Professor Zhang Aimin of Hebei University) are all test strains. The bacterial powder was prepared by the laboratory of Bioengineering Technology Innovation Center of Hebei University. The bacterial powder preparation process is as follows: slant culture → primary shake flask → secondary shake flask → centrifugation at 7000 rpm → collection of bacterial sludge → addition of calcium carbonate to dry (the powder is obtained after crushing) → determination of effective viable bacteria count.

[0058] Mix the bacterial powder with light calcium carbonate → 10 billion or 100 billion / g bacterial powder. Among them, HX-02 bacterial powder is 10 billion / g; RKN1111 and JS bacterial powder are both 100 billion / g.

[0059] The results of the harvest test are shown in Table 2.

[0060]

[0061] As shown in Table 2, treatment T3 (FHF+HX-02+RKN1111) was significantly higher than other treatments. The yield of T3 treatment was 624.86 kg / mu, which was 56.32 kg higher than the yield of 567.09 kg / mu of conventional fertilization T1 treatment, with an increase rate of 10.20%; it was 3.2% higher than the yield of 604.31 kg / mu of T4 with the HX-02 strain alone, and 3.5% higher than the other combinations T2 and T5. Comprehensively considering the yield increase effect and cost, T3 is the best combination.

[0062] Example 3 Effect of Paenibacillus jelly HX-02 on reducing fertilizer usage and increasing corn yield

[0063] The experimental field is located in Zhen Village, Boye County, at 115°32″ east longitude and 38°20″ north latitude. The soil is light loamy tidal brown soil. The experimental field area is 4140.96m 2 The corn variety is Zhengdan 958.

[0064] The treatment groups were set up as follows: T1: 50 kg of conventional corn compound fertilizer (45%), purchased from Baoding Watson Fertilizer Manufacturing Co., Ltd.

[0065] T2: 50 kg of 36% compound microbial fertilizer.

[0066] T3: 50 kg of 38% compound microbial fertilizer.

[0067] Preparation of 38% compound microbial fertilizer: First, the bacterial powder particles were prepared. 12 kg of jelly-like Bacillus HX-02 and Bacillus Velez RKN1111 powder (mass ratio of 5:1) were evenly adhered on the surface of 1000 kg of organic fertilizer particles to obtain bacterial powder particles; then 47.5% (N+P2O5+K2O=47.5% (the mass ratio of N, P2O5 and K2O is 20:10:8) 800 kg of compound fertilizer (FHF: Baoding Watson Fertilizer Manufacturing Co., Ltd.) was mixed with 200 kg of bacterial powder particles in a mass ratio of 4:1 to obtain 38% (N+P2O5+K2O=38%) compound microbial fertilizer.

[0068] Preparation of 36% compound microbial fertilizer: First, the bacterial powder particles were prepared. 12 kg of equal proportions of jelly-like Bacillus and Bacillus Velezii were evenly adhered on the surface of 1000 kg of organic fertilizer particles to obtain bacterial powder particles; then 45.0% (N+P2O5+K2O=45.0%) 800 kg of compound fertilizer (FHF) was mixed with 200 kg of bacterial powder particles in a mass ratio of 4:1 to obtain 36% (N+P2O5+K2O=36%) compound microbial fertilizer. The test results at the harvest period are shown in Tables 3 and 4.

[0069] Table 3 2021 harvest season test seed list

[0070] Table 4 2022 harvest season test seed list

[0071] As can be seen from Tables 3 and 4, the results in 2021 and 2022 are similar. The yield of T3 treatment 624.86 kg / mu is 57.77 kg higher than the yield of conventional fertilization T1 treatment 567.09 kg / mu, with an increase rate of 10.20%. At the same time, it can be seen that treatment T3 is also significantly higher than other treatments. The two-year field test in Boye County compared the effects of different combinations of microbial fertilizers on the growth traits and yield of corn. The results showed that compared with conventional compound fertilizers, compound microbial fertilizers with different strains can significantly improve the growth potential and yield of corn.

[0072] Example 4 Effect of Paenibacillus jelly HX-02 on reducing fertilizer usage and increasing wheat yield

[0073] The composite microbial fertilizer of the present invention was used in a wheat field application test, and also achieved a good yield-increasing effect. The wheat variety was Jingke 938.

[0074] The method of using basal fertilizer was adopted, with four treatments, three replications and random arrangement.

[0075] T1: 50 kg / mu of compound microbial fertilizer as base application; T2: 50 kg / mu of inactivated compound microbial fertilizer as base application; T3: 50 kg / mu of nitrogen, phosphorus and potassium compound fertilizer equal to T1; T4: No use as control.

[0076] Technical indicators of compound microbial fertilizer: N+P2O5+ K2O=38%, effective live bacteria count of HX-02 strain and RKN1111 bacterial powder ≥150 million / g.

[0077] Since wheat was planted in early October 2021, according to the results of field surveys, the application of basic biological organic fertilizer has a significant promoting effect on the growth and development of wheat. The yield survey and seed test results of each plot are shown in Tables 5 and 6.

[0078] Table 5 Yield of each plot

[0079] Table 6 F test table

[0080] F 0.05 (3,6) = 4.757, F 0.05 (2,6) = 5.143. Asterisks indicate significant differences ( p <0.05).

[0081] Table 7 Comparison of yield differences among plots

[0082] LSD 0.05 =1.075, LSD 0.01 =1 .629. Lowercase letters indicate the significant difference between treatments at the 5% level, different letters represent the significant difference level, and the same letters represent the insignificant difference; uppercase letters indicate the significant difference between treatments at the 1% level, different letters represent the extremely significant difference level, and the same letters represent the insignificant difference.

[0083] The yield differences of each plot are compared in Table 7. From the above experimental results, it can be seen that the compound microbial fertilizer of HX-02 strain combined with RKN1111 strain has the best effect at 50 kg / mu. Compared with the control, the yield per mu increased by 44.5 kg, the increase rate was 12.4%, and the difference reached the extremely significant level, thus confirming that the fertilizer has a good yield-increasing effect on wheat.

[0084] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A composite microbial agent, characterized in that: The active ingredient is jelly-like Bacillus ( Paenibacillus mucilaginosus ) HX-02 and Bacillus velezinoffe ( Bacillus velezensis )RKN1111; The deposit number of Paenibacillus jelly HX-02 is CGMCC No.30949; The deposit number of Bacillus velez RKN1111 is CGMCC No.29300.

2. A biological fertilizer or plant growth promoter prepared from the composite microbial agent according to claim 1.

3. A compound microbial fertilizer, characterized in that: It is composed of the composite microbial agent as claimed in claim 1, inorganic fertilizers of nitrogen, phosphorus and potassium, and organic matter; The number of live bacteria of Paenibacillus jelly in the composite microbial fertilizer is 30-70 million / g, and the number of live bacteria of Bacillus velez is 50-150 million / g; The organic matter is selected from one or more of humic acid, bentonite and peat.

4. The composite microbial fertilizer according to claim 3, characterized in that: The nitrogen, phosphorus and potassium inorganic fertilizer is a chemical fertilizer with N+P2O5+K2O=38%.

5. The composite microbial fertilizer according to claim 3, characterized in that: The nitrogen, phosphorus and potassium inorganic fertilizer is prepared by mixing N, P2O5 and K2O in a mass ratio of 20:10:

8.

6. The composite microbial fertilizer according to claim 3, characterized in that: The preparation method of the composite microbial fertilizer comprises: fermenting and culturing the jelly-like Paenibacillus HX-02 and the Velez Bacillus RKN1111 respectively, centrifugally concentrating and drying the fermented liquid to prepare jelly-like Paenibacillus powder and Velez Bacillus powder respectively, then mixing the two powders at a mass ratio of 5:1, and compounding the obtained composite powder with nitrogen, phosphorus and potassium inorganic fertilizers and organic matter; Among them, the mass ratio of composite bacterial powder, nitrogen, phosphorus and potassium inorganic fertilizers and organic matter is 1: (120-145): (20-40).

7. The composite microbial fertilizer according to any one of claims 4 to 6, characterized in that: The formula of the culture medium for fermenting and culturing the jelly-like Paenibacillus is: 15 g starch, 2.0 g (NH4)2SO4, 2.0 g KH2PO4, 1.5 g yeast extract, 0.5 g FeCl3, 1000 mL water, pH 7.5; The formula of the culture medium for fermenting and culturing the Velez subtilis is: 10.0 g starch, 20.0 g bean cake powder, 4.0 g MgSO4, 2.0 g yeast extract, 1000 mL water, pH 7.

0.

8. Use of the composite microbial fertilizer according to any one of claims 3 to 7 in field planting.

9. Use of the composite microbial fertilizer according to any one of claims 3 to 7 in increasing soil fertility and promoting plant growth.

10. The use according to claim 9, characterized in that: The plants include corn and wheat.

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