Multifunctional microbial agent and use thereof

Abstract

A multifunctional microbial agent and the use thereof. The multifunctional microbial agent comprises a composite microbial agent and an adsorbable carrier, wherein the adsorbable carrier comprises livestock and poultry manure, mushroom bran and straw. Three novel Bacillus strains having nitrogen fixation, phosphate solubilizing and potassium solubilizing activities are obtained by means of screening, and serve as excellent microbial agent production strains. The microbial agent produced by means of mixing the composite microbial agent and exogenous organic raw materials the livestock and poultry manure and the crop straw can better improve the microbial environment of soil, activate soil for nutrient retention, and promote nutrient absorption of crops, thus improving the yield and quality of crops.

Description

A multifunctional microbial agent and its application Technical Field

[0001] The present invention relates to the field of bioengineering technology, in particular to a multifunctional microbial agent and application thereof. Background Art

[0002] Microbial inoculants are green, environmentally friendly agricultural products with advantages such as increasing soil fertility, reducing the use of chemical fertilizers and pesticides, purifying and remediating soil, reducing plant diseases, and improving quality and yield. However, microbial growth cycles are long, and some strains have stringent requirements for their growth environment. Multifunctional microbial inoculants combined with adsorbable carriers can better meet the environmental and nutritional needs of microorganisms. Once applied to the soil, the functional bacteria are more likely to colonize and quickly take effect.

[0003] Agricultural waste mainly includes crop straw and livestock and poultry manure. If it enters the environment without proper treatment, it will easily cause waste of resources and environmental pollution.

[0004] Multifunctional microbial agents are primarily made by combining livestock and poultry manure, crop straw, and other materials with microbial liquid. These agents utilize simple strains and are highly effective in promoting plant growth. This can effectively reduce resource waste and environmental pollution caused by agricultural waste.

[0005] Summary of the Invention

[0006] The purpose of the present invention is to provide a multifunctional microbial agent and application thereof.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides a multifunctional microbial agent, comprising a composite microbial bacterial solution and an adsorbable carrier; the adsorbable carrier comprises livestock and poultry excrement, bacterial chaff and straw;

[0009] The composite microbial bacterial liquid comprises: nitrogen-fixing jelly-like Paenibacillus MSSW01, potassium-dissolving jelly-like Paenibacillus MSSW02 and jelly-like Paenibacillus MSSW03.

[0010] Preferably, the volume ratio of the nitrogen-fixing Paenibacillus MSSW01, the potassium-solubilizing Paenibacillus MSSW02 and the Paenibacillus MSSW03 is 1-2:1-2:1-2.

[0011] Preferably, the viable bacterial counts of the nitrogen-fixing jelly-like Paenibacillus MSSW01, potassium-solubilizing jelly-like Paenibacillus MSSW02 and jelly-like Paenibacillus MSSW03 are respectively 2 to 5×10 9 cfu·mL -1 .

[0012] Preferably, the viable bacteria count of the composite microbial solution is 2 to 5×10 9 cfu·mL -1 .

[0013] Preferably, the mass ratio of livestock and poultry manure, mushroom chaff and straw is 25-35:10-20:8-12.

[0014] Preferably, the mass volume ratio of the adsorbable carrier to the multifunctional microbial agent is 3-10 g:1 ml.

[0015] Preferably, the livestock manure is pig manure, cow manure, sheep manure, chicken manure or duck manure; the straw is corn straw; and the moisture content of the adsorbable carrier is 20-25%.

[0016] The present invention also provides application of the multifunctional microbial agent in wheat planting.

[0017] The present invention also provides the use of the multifunctional microbial agent in soil nitrogen fixation, potassium solubilization or phosphorus solubilization.

[0018] This invention screened and identified three new Bacillus strains with nitrogen-fixing, phosphate-solubilizing, and potassium-solubilizing activities, respectively, and used them as excellent microbial agent production strains. The multifunctional microbial agent, produced by mixing the microbial solution with adsorbable carriers—livestock manure and crop straw—can improve the soil microbial environment, activate soil nutrient retention, promote crop nutrient absorption, and increase crop yield and quality.

[0019] Biological Deposit Description

[0020] The nitrogen-fixing jelly-like Paenibacillus MSSW01, whose Latin name is Paenibacillus mucilaginosus, is deposited in the China Center for Type Culture Collection, Wuhan University, Wuhan, China. The deposit date is August 14, 2023, and the deposit number is CCTCC NO: M 20231471.

[0021] Paenibacillus mucilaginosus MSSW02, whose Latin name is Paenibacillus mucilaginosus, is deposited in the China Center for Type Culture Collection, Wuhan University, Wuhan, China. The deposit date is August 14, 2023, and the deposit number is CCTCC NO: M 20231472.

[0022] Paenibacillus mucilaginosus MSSW03;

[0023] The strain was deposited in the China Center for Type Culture Collection: address: Wuhan University, the deposit date is August 14, 2023, and the deposit number is CCTCC NO: M 20231473. DETAILED DESCRIPTION

[0024] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0025] Example 1

[0026] A method for preparing a multifunctional microbial agent comprises the following steps:

[0027] (1) Bacillus jelly MSSW03 (viable count 3×10 9 cfu·mL -1 ), Paenibacillus sp. MSSW02 (viable count 3×10 9 cfu·mL -1 ), Paenibacillus nitrogenus MSSW01 (viable count 3×10 9 cfu·mL -1 ) The bacterial solutions of the three strains were mixed in a volume ratio of 1:1:1 to obtain a multifunctional microbial agent (viable bacteria count 3×10 9 cfu·mL -1 ).

[0028] (2) Pig manure, fungus dregs and crop straw are mixed in a mass ratio of 30:15:10 and placed in a mixing machine, and the water content is adjusted to 23% to obtain a mixture as an adsorbable carrier. Then, the multifunctional microbial agent obtained in step (1) is added in a mass volume ratio of 7g:1ml, and the mixture is fully stirred to obtain a multifunctional microbial agent product. The number of viable bacteria in the multifunctional microbial agent is not less than 5×10 8 cfu·mL -1 .

[0029] Example 2

[0030] (1) Taking wheat as an example, the growth-promoting effect of multifunctional microbial agents on crops was explored.

[0031] In a 10cm long and wide plastic pot, add farmland soil and compact it to 1.5cm from the pot rim. Evenly place 25 wheat seeds of roughly the same size and cover with soil up to the pot rim. Add an appropriate amount of water to maintain a consistent soil moisture level in each pot, and water each pot the same amount daily. Water each pot every two days for the next step.

[0032] One week after emergence, the multifunctional microbial agent in Example 5 was inoculated by watering. Specifically, 10 g of the multifunctional microbial agent was accurately weighed and dissolved in 90 mL of sterile saline. 1 mL of the solution was drawn, diluted with 9 mL of sterile saline and applied, and 5 mL was inoculated into each flower pot. Five replicates were set for each treatment. The potted plants not inoculated with the agent were used as the blank control group ck, and other commercially available multifunctional microbial agent products (Guangxi Nongbao Bioengineering Co., Ltd.; microbial agent (agricultural special) Bacillus subtilis) were used as the positive control group.

[0033] Cultivation conditions: Cultivate outdoors, irrigate with distilled water, and harvest after 30 days.

[0034] 30 days after sowing, carefully pull out the entire wheat plant in the pot, shake off most of the soil stuck to the roots, wash it with tap water and dry it with newspaper, spread it flat on the table one by one, and use a ruler to measure the length from the cotyledon node to the growth point as the wheat plant height.

[0035] Use a ruler to measure the length of the third true cotyledon as the leaf length of wheat.

[0036] The wheat plants were washed and dried, and the number of roots extending from the embryo of the wheat seeds was counted as the number of seed roots.

[0037] The maximum root length was measured using a ruler from the junction of the wheat rhizome to the longest root tip of the entire plant, that is, the length of the longest seed root in the seed embryo.

[0038] After washing and drying the wheat plants, they were weighed directly using a balance (accurate to 0.01 g) to obtain their fresh weight.

[0039] The wheat plants were then dried in an oven at 100°C until constant weight was reached and weighed to determine their dry weight. The above data were recorded and shown in Table 1.

[0040] Table 1 Effects of different treatments on wheat traits

[0041] As shown in Table 1, the plant height, leaf length, maximum root length, fresh weight, and dry weight of wheat inoculated with the multifunctional microbial agent produced by the present invention were significantly increased compared to the blank control group, increasing by 17.75%, 42.27%, 33.03%, 55.79%, and 133.33%, respectively. Compared with other commercially available multifunctional microbial agents, these increases were 1.17%, 2.22%, 9.85%, 2.70%, and 9.30%, respectively. The multifunctional microbial agent produced by the present invention has a significant effect on promoting wheat growth and has great market potential.

[0042] (2) Experiments on determination of soil physical and chemical properties

[0043] Thirty days after sowing and harvesting in step (1), the potted soil from each experimental group was collected, air-dried, sieved, and packaged into plastic bags. pH, organic matter, total nitrogen, available potassium, and available phosphorus were measured. Soil that had not been planted with wheat was used as the original soil group. The data are recorded in Table 2.

[0044] Determination of total nitrogen content in soil: Refer to NY / T 53-1987, "Determination of Total Nitrogen in Soil (Semi-micro Kelvin Method)." In the presence of sodium thiosulfate, concentrated sulfuric acid, perchloric acid, and a catalyst, a redox reaction converts all nitrogen into ammonium. The digested solution is alkalized, and the distilled ammonia is absorbed by boric acid. Titration is then performed with standard hydrochloric acid solution, and the total nitrogen content in the soil is calculated based on the amount of standard hydrochloric acid solution used.

[0045] Determination of available potassium in soil: Refer to NY / T 889-2004, "Determination of available and slow-release potassium in soil." Weigh 5 g of air-dried soil sample that has passed a 1-µm sieve into a 100-mL Erlenmeyer flask. Add 50.0 mL of ammonium acetate solution (soil-to-liquid ratio of 1:10). Tightly cap the flask, shake at 150-180 rpm for 30 min at 20-25°C, and dry filter. Measure the filtrate directly using a flame photometer.

[0046] Determination of available phosphorus in soil: Referencing NY / T 1121.7-2014, "Soil Testing Part 7: Determination of Available Phosphorus in Soil," extract the available phosphorus in the soil using a 0.5 mol / L sodium bicarbonate solution (pH 8.5). The phosphorus in the extract reacts with a molybdenum antimony anti-colorant to produce phosphomolybdenum blue, the absorbance of which is measured at a wavelength of 880 nm. Within a certain concentration range, the phosphorus content and absorbance values ​​conform to the Lambert-Beer law.

[0047] Table 2 Effects of applying multifunctional microbial agents on soil fertility

[0048] The experimental results are shown in Table 2. After the experiment in step (1), the potted soil was tested for indicators. It was found that the nutritional indicators of the soil in which the multifunctional microbial agent was applied were greatly improved. Compared with the blank control group, the total nitrogen content increased by 291.20%, the available phosphorus content increased by 298.51%, the available potassium content increased by 283.71%, and the organic matter content increased by 36.68%. Compared with other commercially available multifunctional microbial agents, they increased by 1.45%, 2.02%, 1.38%, and 4.36%, respectively.

[0049] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A multifunctional microbial inoculum, characterized in that, It includes a composite microbial agent and an adsorbable carrier; the adsorbable carrier includes livestock and poultry manure, mushroom residue, and straw; The composite microbial agent includes: Paenibacillus mucilaginosus MSSW01 for nitrogen fixation, Paenibacillus mucilaginosus MSSW02 for potassium solubilization, and Paenibacillus mucilaginosus MSSW03; The preservation number of Paenibacillus mucilaginosus MSSW01 for nitrogen fixation is CCTCC NO: M 20231471; The preservation number of Paenibacillus mucilaginosus MSSW02 for potassium solubilization is CCTCC NO: M 20231472; The preservation number of Paenibacillus mucilaginosus MSSW03 is CCTCC NO: M 20231473; The volume ratio of Paenibacillus mucilaginosus MSSW01 for nitrogen fixation, Paenibacillus mucilaginosus MSSW02 for potassium solubilization, and Paenibacillus mucilaginosus MSSW03 is 1-2:1-2:1-2.

2. The multifunctional microbial inoculant according to claim 1, characterized in that, The types of the livestock and poultry manure are pig manure, cow manure, sheep manure, chicken manure, or duck manure.

3. The multifunctional microbial inoculum according to claim 1, wherein The viable counts of the nitrogen-fixing Paenibacillus mucilaginosus MSSW01, potassium-solubilizing Paenibacillus mucilaginosus MSSW02, and Paenibacillus mucilaginosus MSSW03 are respectively: 2 - 5×10 9 cfu·mL -1 .

4. The multifunctional microbial inoculant according to claim 1, characterized in that, The viable count of the compound bacterial agent is 2-5×10 9 cfu·mL -1 .

5. The multifunctional microbial inoculum according to claim 1, characterized in that, The mass ratio of the livestock and poultry manure, mushroom residue, and straw is 25-35:10-20:8-12.

6. The multifunctional microbial inoculant according to claim 1, wherein The mass-volume ratio of the adsorbable carrier to the composite microbial agent is 3-10 g:1 ml.

7. The multifunctional microbial inoculant according to claim 1, characterized in that, The type of the straw is corn straw; the water content of the adsorbable carrier is 20-25%.

8. Application of the multifunctional microbial agent according to any one of claims 1-7 in wheat planting.

9. Application of the multifunctional microbial agent according to any one of claims 1-7 in soil nitrogen fixation, potassium solubilization, or phosphorus solubilization.

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