Soil remediation and water retention microbial fertilizer

Abstract

The invention relates to a soil remediation and water retention microbial fertilizer, which is prepared by culturing a compound strain and optimizing nutrient substances in a culture dish, preparing the most suitable growth environment of the mixed strain, increasing the effective viable count of a microbial agent, and mixing the mixed agent with organic and inorganic fertilizers to form the compound microbial fertilizer with good water retention.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial fertilizers, and in particular relates to a soil remediation and water-retaining microbial fertilizer. Background Art

[0002] In the development of the national economy, agriculture occupies an unshakable and important position. Over the years, my country's grain production has made breakthrough progress. One of the factors is the massive promotion of chemical fertilizers. However, the long-term excessive use of chemical fertilizers has also brought about a series of problems such as the decline in the quality of agricultural products, soil compaction, environmental pollution, and ecological deterioration, which has led to an increase in agricultural costs and affected ecological benefits. If this problem cannot be solved, it will not only affect the sustainable development between agricultural resources and the environment, but also reduce the quality of cultivated land and affect the sustainable development of agriculture. Compared with traditional chemical fertilizers, microbial fertilizers are a new type of fertilizer that is green, safe, economical, and environmentally friendly. It can effectively improve the utilization rate of chemical fertilizers, promote pesticide absorption, reduce crop diseases, and protect the farmland environment. Under the influence of soil pollution, climate change, and unreasonable fertilization, vigorously using microbial fertilizers to improve the quality of cultivated land is a key issue of current concern.

[0003] The single microbial fertilization effects of ammonia-fixing bacteria, silicates, phosphate-solubilizing fungi, etc. are relatively simple, while the composite microbial fertilization system made by mixed bacterial culture and optimized culture environment can not only supply nitrogen, phosphorus, potassium and other nutrients to crops, but also effectively improve the microbial environment of the land. This study uses microorganisms, organic components, and inorganic components as composite components to form an integrated composite microbial fertilizer product, so that the product can achieve the function of "one fertilizer with multiple effects" when applied to the soil. On the one hand, it can provide the fast-acting N, P, and K required for crop growth, so as to achieve the effect of increasing production and income with less or no chemical fertilizers; on the other hand, it can promote crop growth, improve the nutritional conditions of crops, and promote the development of crop root systems; at the same time, it can also increase the beneficial microbial flora in the soil, improve soil fertility, and improve the nutrient supply of crops and the quality of crop production, thereby saving land use costs and effectively repairing and protecting soil structure. Summary of the invention

[0004] The purpose of the invention is to provide a soil repair and water-retaining microbial fertilizer.

[0005] In order to solve the above technical problems, the specific process of the present invention is as follows:

[0006] A soil remediation and water-retaining microbial fertilizer, wherein the culture process of the composite fermentation agent is as follows: Bacillus licheniformis, Bacillus megaterium, and Pseudomonas aeruginosa are inoculated into LB liquid culture medium respectively, and cultured at 180-220 r / min shaking table and 37°C for 16-24 hours to obtain Bacillus licheniformis, Bacillus megaterium, and Pseudomonas aeruginosa seed liquid; LB liquid culture medium: 8-12 g / L peptone, 8-12 g / L NaCl, 5 g / L yeast extract powder, 2% agar, ammonia water to adjust pH to 7.2-7.4, and distilled water to 1 L;

[0007] Inoculate 2% of Bacillus licheniformis, 2% of Bacillus megaterium and 2% of Pseudomonas aeruginosa seed solution in a mass ratio into an optimized culture medium, ferment at 30°C, 140-160r / min for 28-36h to obtain a composite fermentation agent, and store it at 4°C for later use.

[0008] Among them, the optimized culture medium is: 8-12g / L carbon source, 13-14g / L soybean meal powder, 2-3g / L ammonium chloride, 8-12g / L yeast extract, 3-5g / L dipotassium hydrogen phosphate, 5-7g / L potassium dihydrogen phosphate and 0.8-1.0g / L manganese sulfate.

[0009] Wherein, the carbon source is D-erythrose.

[0010] A soil remediation and water-retaining microbial fertilizer further comprises the following steps:

[0011] 24-28 parts of urea solution with a mass fraction of 20-25%, 10-15 parts of monoammonium phosphate solution with a mass fraction of 8-12%, 7-11 parts of potassium sulfate solution with a mass fraction of 6-9%, 0.6-1.0 parts of sodium borate solution with a mass fraction of 10-20%, 50-70 parts of dry chicken manure, 8-12 parts of white clay, and 0.9-1.4 parts of potassium L-aspartate solution with a mass fraction of 8-15% are mixed, and 6-9% of a composite fermentation bacterial agent is added, and the mixture is stirred at 800-1500 r / min for 40-60 minutes to be evenly mixed, and air-dried to control the water content to be below 20%.

[0012] The beneficial effects of the present invention are:

[0013] 1. By cultivating the composite strain and optimizing the nutrients in the culture dish, the most suitable growth environment for the composite strain is prepared, the effective live bacteria of the microbial agent is increased, and the composite bacteria is mixed with organic and inorganic fertilizers to form a composite microbial fertilizer with good water retention. The composite microbial fertilizer changes the salinity and alkali components in the soil through the physiological activities of microorganisms, reduces the concentration and content of salinity, improves soil fertility, improves crop nutrition supply and crop production quality, thereby saving land use costs and effectively repairing and protecting soil structure.

[0014] 2. The mixed microbial flora is composed of three mixed strains of Bacillus licheniformis, Bacillus megaterium, and Pseudomonas aeruginosa. The mixed strains have no antagonism to each other and can inhibit harmful bacteria such as Escherichia coli and Staphylococcus aureus during the growth and reproduction process. They also have good phosphorus, potassium and nitrogen-solubilizing abilities and promote plant growth.

[0015] 3. The carbon source in the mixed strain culture dish of the present invention is D-erythrose, which is a four-carbon sugar with a simpler structure than glucose (hexose). Although glucose is a common carbon source, the carbon skeleton of D-erythrose is smaller, and it can participate in the metabolic pathway more quickly in the microbial metabolism process, and its smaller molecular structure may enable microorganisms to utilize and convert more quickly. This simple structure can be more efficiently metabolized and utilized by some microorganisms, providing more energy to support the growth of microorganisms. Using D-erythrose as a carbon source in compound microbial fertilizers can selectively promote the growth of beneficial bacteria and reduce the competition of some unfavorable microorganisms. In addition, the organic acids or other metabolites that may be generated by D-erythrose during the metabolism process have a certain inhibitory effect on the community balance of microorganisms and harmful bacteria in the soil (such as Escherichia coli and Staphylococcus aureus), thereby enhancing the disease resistance and environmental adaptability of compound microbial fertilizers. D-erythrose can effectively promote its nitrogen fixation activity in the metabolic process of some nitrogen-fixing microorganisms. Studies have shown that some nitrogen-fixing bacteria, such as some Bacillus, can use D-erythrose as a carbon source to provide energy, thereby showing higher efficiency in fixing nitrogen in the atmosphere. In contrast, glucose may cause some microorganisms to produce more byproducts, affecting their nitrogen fixation efficiency, while the simplified metabolic pathway of D-erythrose helps to improve nitrogen fixation ability.

[0016] 4. The organic fertilizer composition of the present invention makes full use of the high nutrient content of chicken manure and its easy availability. Long-term application can provide sufficient nutrients for crops, make the root and stem of crops develop well, and enhance the cold and frost resistance of crops. In addition, the manure is relatively sticky and heavy, and when applied to the soil, it has better water retention and improvement and repair effects on sandy land that has leaked fertilizer and water. DETAILED DESCRIPTION

[0017] The present invention is further described in detail below with reference to the embodiments.

[0018] Example 1

[0019] Cultivation of composite fermentation agent: Bacillus licheniformis, Bacillus megaterium, and Pseudomonas aeruginosa were inoculated into LB liquid culture medium respectively, and cultured at 200 r / min shaking table and 37° C. for 20 h to obtain seed liquids of Bacillus licheniformis, Bacillus megaterium, and Pseudomonas aeruginosa; LB liquid culture medium: 10 g / L peptone, 10 g / L NaCl, 5 g / L yeast extract powder, 2% agar, ammonia water to adjust pH to 7.3, and distilled water to 1 L;

[0020] The seed solution of Bacillus licheniformis, Bacillus megaterium and Pseudomonas aeruginosa with a mass ratio of 2% was inoculated into the optimized culture medium, and fermented at 30°C, 150r / min for 32h to obtain a composite fermentation agent, which was stored at 4°C for later use; the optimized culture medium was: 10g / L D-erythrose, 13.5g / L soybean meal powder, 2.5g / L ammonium chloride, 10g / L yeast extract, 4g / L dipotassium hydrogen phosphate, 6g / L potassium dihydrogen phosphate and 0.9g / L manganese sulfate.

[0021] 12 parts by mass of 10% monoammonium phosphate solution, 9 parts by mass of 7.5% potassium sulfate solution, 0.8 parts by mass of 15% sodium borate solution, 60 parts by mass of dry chicken manure, 10 parts by mass of white clay, and 1.2 parts by mass of 12% potassium L-aspartate solution are mixed, and 7.5% of a composite fermentation agent is added, and the mixture is stirred at 1300 r / min for 50 minutes to be evenly mixed, and air-dried to control the water content to be below 20%.

[0022] Example 2

[0023] Cultivation of composite fermentation agent: Bacillus licheniformis, Bacillus megaterium, and Pseudomonas aeruginosa were inoculated into LB liquid culture medium respectively, and cultured at 220 r / min shaking table and 37° C. for 16 h to obtain seed liquids of Bacillus licheniformis, Bacillus megaterium, and Pseudomonas aeruginosa; LB liquid culture medium: 12 g / L peptone, 8 g / L NaCl, 5 g / L yeast extract powder, 2% agar, ammonia water to adjust pH to 7.4, and distilled water to 1 L;

[0024] The seed solution of Bacillus licheniformis, Bacillus megaterium and Pseudomonas aeruginosa with a mass ratio of 2% is inoculated into the optimized culture medium, and fermented at 30°C, 140r / min for 36 hours to obtain a composite fermentation agent, which is stored at 4°C for later use; the optimized culture medium is: 8g / L D-erythrose, 14g / L soybean meal powder, 2g / L ammonium chloride, 12g / L yeast extract, 3g / L dipotassium hydrogen phosphate, 7g / L potassium dihydrogen phosphate and 0.8g / L manganese sulfate.

[0025] Mix 15 parts of 12% monoammonium phosphate solution, 7 parts of 9% potassium sulfate solution, 1.0 part of 10% sodium borate solution, 70 parts of dry chicken manure, 12 parts of white clay, and 0.9 part of 15% potassium L-aspartate solution, add 9% composite fermentation bacteria agent, stir at 1500r / min for 40min to mix evenly, and air-dry to control the water content below 20%.

[0026] Example 3

[0027] Cultivation of composite fermentation agent: Bacillus licheniformis, Bacillus megaterium, and Pseudomonas aeruginosa were inoculated into LB liquid culture medium respectively, and cultured at 180 r / min shaking table and 37° C. for 24 h to obtain seed liquids of Bacillus licheniformis, Bacillus megaterium, and Pseudomonas aeruginosa; LB liquid culture medium: 8 g / L peptone, 12 g / L NaCl, 5 g / L yeast extract powder, 2% agar, ammonia water to adjust pH to 7.2, and distilled water to 1 L;

[0028] The seed solution of Bacillus licheniformis, Bacillus megaterium and Pseudomonas aeruginosa with a mass ratio of 2% was inoculated into the optimized culture medium, and fermented at 30°C, 160r / min for 28h to obtain a composite fermentation agent, which was stored at 4°C for later use; the optimized culture medium was: 12g / L D-erythrose, 13g / L soybean meal powder, 3g / L ammonium chloride, 8g / L yeast extract, 5g / L dipotassium hydrogen phosphate, 5g / L potassium dihydrogen phosphate and 1.0g / L manganese sulfate.

[0029] 10 parts by mass of 8-12% monoammonium phosphate solution, 11 parts by mass of 6% potassium sulfate solution, 0.6 parts by mass of 20% sodium borate solution, 50 parts by mass of dry chicken manure, 8 parts by mass of white clay, and 1.4 parts by mass of 8% potassium L-aspartate solution are mixed, and 6% of a composite fermentation agent is added, and the mixture is stirred at 800 r / min for 60 minutes to be evenly mixed, and air-dried to control the water content to be below 20%.

[0030] Comparative Example 1

[0031] The difference between this comparative example and Example 1 is that the fermentation agent used in this comparative example is a single agent of Bacillus licheniformis, and the rest is the same as Example 1.

[0032] Comparative Example 2

[0033] The difference between this comparative example and Example 1 is that the fermentation bacteria used in this comparative example are Bacillus licheniformis and Bacillus megaterium, and the rest is the same as Example 1.

[0034] Comparative Example 3

[0035] The difference between this comparative example and Example 1 is that the fermentation bacteria used in this comparative example are Bacillus amyloliquefaciens, Bacillus megaterium and Pseudomonas aeruginosa, and the rest is the same as Example 1.

[0036] Comparative Example 4

[0037] The difference between this comparative example and Example 1 is that the fermentation bacteria used in this comparative example are Bacillus licheniformis, Bacillus megaterium and Bacillus amyloliquefaciens, and the rest is the same as Example 1.

[0038] Comparative Example 5

[0039] The difference between this comparative example and Example 1 is that the fermentation bacteria used in this comparative example are Bacillus licheniformis, Bacillus cereus and Pseudomonas aeruginosa, and the rest is the same as Example 1.

[0040] Comparative Example 6

[0041] The difference between this comparative example and Example 1 is that D-erythrose in this comparative example is glucose; the rest is the same as Example 1.

[0042] Comparative Example 7

[0043] The difference between this comparative example and Example 1 is that the amount of D-erythrose added in this comparative example is different, wherein the optimized culture medium is: 16 g / L D-erythrose, 13.5 g / L soybean meal powder, 2.5 g / L ammonium chloride, 10 g / L yeast extract, 4 g / L dipotassium hydrogen phosphate, 6 g / L potassium dihydrogen phosphate and 0.9 g / L manganese sulfate; the rest is the same as Example 1.

[0044] Comparative Example 8

[0045] The difference between this comparative example and Example 1 is that the amount of D-erythrose added in this comparative example is different, wherein the optimized culture medium is: 5g / L D-erythrose, 13.5g / L soybean meal powder, 2.5g / L ammonium chloride, 10g / L yeast extract, 4g / L dipotassium hydrogen phosphate, 6g / L potassium dihydrogen phosphate and 0.9g / L manganese sulfate; the rest is the same as Example 1.

[0046] Comparative Example 9

[0047] The difference between this comparative example and Example 1 is that the chicken manure in this comparative example is humus, and the rest is the same as Example 1.

[0048] Experiment 1: Effect of culture environment on bacterial activity

[0049] The fermentation bacteria obtained above were tested for bacterial activity, and the results are shown in Table 1.

[0050] Table 1

[0051] sample Inoculated viable count (cfu / mL) Viable bacteria count after culture (cfu / mL) Example 1 <![CDATA[8.72×10 7 ]]> <![CDATA[2.82×10 9 ]]> Example 2 <![CDATA[8.56×10 7 ]]> <![CDATA[2.74×10 9 ]]> Example 3 <![CDATA[8.28×10 7 ]]> <![CDATA[2.58×10 9 ]]> Comparative Example 1 <![CDATA[4.53×10 6 ]]> <![CDATA[9.67×10 7 ]]> Comparative Example 2 <![CDATA[7.87×10 6 ]]> <![CDATA[1.68×10 8 ]]> Comparative Example 3 <![CDATA[1.74×10 7 ]]> <![CDATA[2.23×10 8 ]]> Comparative Example 4 <![CDATA[1.82×10 7 ]]> <![CDATA[1.98×10 8 ]]> Comparative Example 5 <![CDATA[1.96×10 7 ]]> <![CDATA[2.10×10 8 ]]> Comparative Example 6 <![CDATA[2.56×10 7 ]]> <![CDATA[5.94×10 8 ]]> Comparative Example 7 <![CDATA[7.24×10 7 ]]> <![CDATA[1.87×10 9 ]]> Comparative Example 8 <![CDATA[5.82×10 7 ]]> <![CDATA[8.64×10 8 ]]>

[0052] Experiment 2: Effect of fertilizer on tomato biomass

[0053] Under the same cultivation conditions, the experiment was set up. The control group was a commercially available compound microbial fertilizer, which was Shangguyuanwo compound microbial fertilizer. 100kg / mu of compound microbial fertilizer was applied at the tomato seedling stage, flowering stage, and fruiting stage. Then the biomass, yield, and soil basic nutrients of tomatoes under different treatments were tested. The test crop was tomato, the test variety was 1612, and the planting method was film drip irrigation, with a configuration of 1 film, 2 tubes, and 2 rows, a plant spacing of 40cm, a row spacing of 60cm, and a theoretical plant number of 5.2*104 plants·hm2 -2 The total irrigation volume was 150 m3, with 8 irrigations and 6 fertilizer applications.

[0054] Determination of tomato biomass: After the tomato flowers first bloom, dig 1m*1m=1m 2 The plants were quickly brought back to the laboratory to measure the fresh biomass of tomatoes, then they were sterilized at 105℃ for 30min and dried at 70℃ to constant weight, and the dry biomass was measured. The results are shown in Table 2. The soil before and after planting was collected to measure the contents of ammonium nitrogen, available phosphorus and quick-acting potassium. The results are shown in Table 3.

[0055] Table 2

[0056] sample Fresh biomass (kg / mu) Dry biomass (kg / mu) Control group 459.95 286.55 Example 1 574.69 363.79 Example 2 572.15 357.02 Example 3 569.86 361.86 Comparative Example 1 464.32 290.2 Comparative Example 2 475.92 301.73 Comparative Example 3 489.28 308.73 Comparative Example 4 496.64 314.87 Comparative Example 5 493.57 311.94 Comparative Example 6 513.68 324.65 Comparative Example 7 558.42 354.59 Comparative Example 8 532.68 332.39 Comparative Example 9 541.59 342.28

[0057] Table 3

[0058]

[0059]

Claims

1. A soil remediation and water-retaining microbial fertilizer, characterized in that: The culture process of the composite fermentation agent is as follows: Bacillus licheniformis, Bacillus megaterium and Pseudomonas aeruginosa are inoculated into LB liquid culture medium respectively, and cultured at 180-220 r / min shaking table and 37° C. for 16-24 hours to obtain seed liquids of Bacillus licheniformis, Bacillus megaterium and Pseudomonas aeruginosa; LB liquid culture medium: 8-12 g / L peptone, 8-12 g / L NaCl, 5 g / L yeast extract powder, 2% agar, ammonia water to adjust pH to 7.2-7.4, and distilled water to 1 L; Inoculate 2% of Bacillus licheniformis, 2% of Bacillus megaterium and 2% of Pseudomonas aeruginosa seed solution in a mass ratio into an optimized culture medium, ferment at 30°C, 140-160r / min for 28-36h to obtain a composite fermentation agent, and store it at 4°C for later use.

2. A soil remediation and water-retaining microbial fertilizer according to claim 1, characterized in that: The optimized culture medium comprises: 8-12 g / L carbon source, 13-14 g / L soybean meal powder, 2-3 g / L ammonium chloride, 8-12 g / L yeast extract, 3-5 g / L dipotassium hydrogen phosphate, 5-7 g / L potassium dihydrogen phosphate and 0.8-1.0 g / L manganese sulfate.

3. A soil remediation and water-retaining microbial fertilizer as claimed in claim 2, characterized in that: The carbon source is D-erythrose.

4. The soil remediation and water-retaining microbial fertilizer according to claim 1, characterized in that: The microbial fertilizer also includes: 24-28 parts of urea solution with a mass fraction of 20-25%, 10-15 parts of monoammonium phosphate solution with a mass fraction of 8-12%, 7-11 parts of potassium sulfate solution with a mass fraction of 6-9%, 0.6-1.0 parts of sodium borate solution with a mass fraction of 10-20%, 50-70 parts of dry chicken manure, 8-12 parts of white clay, 0.9-1.4 parts of L-aspartic acid potassium solution with a mass fraction of 8-15% are mixed, and 6-9% of a composite fermentation bacterial agent is added, and the mixture is stirred at 800-1500r / min for 40-60min to be evenly mixed, and air-dried to control the water content to be below 20%.

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