Complex microbial inoculant for saline-alkali soil and application of complex microbial inoculant

Abstract

The invention relates to a complex microbial inoculant for saline-alkali soil and application of the complex microbial inoculant. Three new low-temperature-resistant strains which respectively have the functions of fixing nitrogen, dissolving inorganic phosphorus, resisting salt and alkali and generating auxin indoleacetic acid are selected and used as excellent microbial agent production strains. The three microbial strains are mixed according to a certain proportion to produce the compound microbial agent, so that the soil microbial environment can be better improved, the soil is activated, nutrients are immobilized, nutrient absorption of crops is promoted, and the yield and quality of the crops are improved.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a composite bacterial agent for saline-alkali land and application thereof. Background Art

[0002] The salt content of light saline-alkali land is less than 0.3%, and the pH value is 7.1-8.5. There are about 99.13 million hectares of saline-alkali land in my country, which are mainly divided into the northwest inland saline-alkali area, the northeast Songnen Plain saline-alkali area, the coastal saline-alkali area, the Huanghuaihai Plain saline-alkali area, and the upper and middle reaches of the Yellow River irrigation area saline-alkali area. At present, my country has achieved initial results in screening salt-alkali tolerant microorganisms, developing efficient microbial preparation technology, and applying microbial agents to improve saline-alkali land soil. However, my country's saline-alkali areas are mainly in the north, with a long low temperature period. The activity of commonly used functional microorganisms decreases in the low temperature season, and their functionality is greatly reduced.

[0003] Psychotropic microorganisms can grow and reproduce normally in low temperature environments. These microorganisms perform well in the low temperature environment of the north and can maintain high biological activity and metabolic capacity under low temperature conditions. They are more suitable for the cultivation of crops in saline-alkali lands in high latitudes. Summary of the invention

[0004] In view of the problems existing in the prior art, the present invention discloses a composite bacterial agent for saline-alkali land, comprising Pseudomonas mohnii TWS2N1, Pseudomonas rustica TWS23 and Penicillium oxalicum TWSF1, wherein the Pseudomonas mohnii TWS2N1 has the functions of fixing nitrogen and producing the plant growth hormone indoleacetic acid (IAA), the Pseudomonas rustica TWS23 has the functions of dissolving inorganic phosphorus and producing the plant growth hormone indoleacetic acid (IAA), and the Penicillium oxalicum TWSF1 is salt-alkali resistant and can dissolve inorganic phosphorus.

[0005] The Pseudomonas mohnii TWS2N1 was deposited in the General Microbiology Center of China National Microorganism Culture Collection on October 25, 2024, with a deposit number of CGMCC No. 32348. The Pseudomonas rustica TWS23 was deposited in the General Microbiology Center of China National Microorganism Culture Collection on September 25, 2024, with a deposit number of CGMCC No. 32078. The Penicillium oxalicum TWSF1 was deposited in the General Microbiology Center of China National Microorganism Culture Collection on October 25, 2024, with a deposit number of CGMCC No. 41553.

[0006] As a preferred embodiment of the present invention, the ratio of the effective viable counts of Pseudomonas mohnii TWS2N1, Pseudomonas rustica TWS23 and Penicillium oxalicum TWSF1 is 1-2:1-2:1-2, and the total effective viable count of the composite bacterial agent is not less than 2.0×10 8 CFU / g.

[0007] As a preferred embodiment of the present invention, the composite bacterial agent can be processed into tablets, granules or powders.

[0008] A method for preparing a composite bacterial agent for saline-alkali land comprises the following steps: Step 1, activating and fermenting the bacterial strains of Pseudomonas mohnii TWS2N1 and Pseudomonas rustica TWS23 respectively, and centrifuging to obtain bacterial sludge of the two bacteria; Step 2: mixing the bacterial sludge of the two bacteria with the adsorbent respectively, and freeze-drying them to obtain two initial bacterial agents; Step 3, activating the fungal strain of Penicillium oxalicum TWSF1 and solid fermenting and culturing it to obtain a fungal culture; Step 4: washing the fungal culture with a Tween-80 solution, filtering the washing solution obtained by washing to obtain a spore suspension, centrifuging the spore suspension, and then precipitating and freeze-drying to obtain fungal spore powder; Step 5, testing the effective viable count of the two initial bacterial inoculants and fungal spore powder; Step 6: According to the total effective live bacteria count of the composite bacterial agent, two initial bacterial agents and fungal spore powder are added in proportion, and the remaining part is filled with an adsorbent, and mixed evenly to obtain the composite bacterial agent.

[0009] The above-mentioned composite bacterial agent can be applied to the cultivation of crops in slightly saline-alkali lands in the cold northern regions.

[0010] The above-mentioned composite bacterial agent can be used in soil nitrogen fixation, phosphorus solubilization and salinization improvement.

[0011] Beneficial effects of the present invention: The present invention selects three new low-temperature resistant strains that respectively have the functions of nitrogen fixation, inorganic phosphorus dissolution, salt and alkali tolerance, and production of plant growth hormone indoleacetic acid as excellent microbial agent production strains. The three microbial strains are mixed in a certain proportion to produce a composite microbial agent, which can better improve the soil microbial environment, activate the soil, fix nutrients, promote crop nutrient absorption, and improve crop yield and quality. DETAILED DESCRIPTION

[0012] Example 1

[0013] The composite bacterial agent for saline-alkali land of the present invention comprises Pseudomonas mohnii TWS2N1, Pseudomonas rustica TWS23 and Penicillium oxalicum TWSF1, and the effective live bacteria count ratio is 1:1:1, and the total effective live bacteria count of the composite bacterial agent is not less than 2.0×10 8 CFU / g.

[0014] Among them, the Pseudomonas mohnii TWS2N1 has the function of fixing nitrogen and producing the plant growth hormone indoleacetic acid (IAA), the Pseudomonas rustica TWS23 has the function of dissolving inorganic phosphorus and producing the plant growth hormone indoleacetic acid (IAA), and the Penicillium oxalicum TWSF1 is salt-alkali resistant and can dissolve inorganic phosphorus.

[0015] A method for preparing a composite bacterial agent for saline-alkali land comprises the following steps: Step 1, activating and fermenting the bacterial strains of Pseudomonas mohnii TWS2N1 and Pseudomonas rustica TWS23 respectively, and centrifuging to obtain bacterial sludge of the two bacteria; Step 2: mixing the bacterial sludge of the two bacteria with the adsorbent respectively, and freeze-drying them to obtain two initial bacterial agents; Step 3, activating the fungal strain of Penicillium oxalicum TWSF1 and solid fermenting and culturing it to obtain a fungal culture; Step 4: washing the fungal culture with a Tween-80 solution, filtering the washing solution obtained by washing to obtain a spore suspension, centrifuging the spore suspension, and then precipitating and freeze-drying to obtain fungal spore powder; Step 5, testing the effective viable count of the two initial bacterial inoculants and fungal spore powder; Step 6: According to the total effective live bacteria count of the composite bacterial agent, two initial bacterial agents and fungal spore powder are added in proportion, and the remaining part is filled with an adsorbent, and mixed evenly to obtain the composite bacterial agent.

[0016] The composite bacterial agent can be processed into tablets, granules or powders, and applied to the planting of crops in slightly saline-alkali lands in cold northern regions, and can also be applied to nitrogen fixation, phosphorus solubilization and salinization improvement of soils.

[0017] Example 2

[0018] The composite bacterial agent for saline-alkali land of the present invention comprises Pseudomonas mohnii TWS2N1, Pseudomonas rustica TWS23 and Penicillium oxalicum TWSF1, and the effective live bacteria count ratio is 2:1:2, and the total effective live bacteria count of the composite bacterial agent is not less than 2.0×10 8 CFU / g.

[0019] Among them, the Pseudomonas mohnii TWS2N1 has the function of fixing nitrogen and producing the plant growth hormone indoleacetic acid (IAA), the Pseudomonas rustica TWS23 has the function of dissolving inorganic phosphorus and producing the plant growth hormone indoleacetic acid (IAA), and the Penicillium oxalicum TWSF1 is salt-alkali resistant and can dissolve inorganic phosphorus.

[0020] A method for preparing a composite bacterial agent for saline-alkali land comprises the following steps: Step 1, activating and fermenting the bacterial strains of Pseudomonas mohnii TWS2N1 and Pseudomonas rustica TWS23 respectively, and centrifuging to obtain bacterial sludge of the two bacteria; Step 2: mixing the bacterial sludge of the two bacteria with the adsorbent respectively, and freeze-drying them to obtain two initial bacterial agents; Step 3, activating the fungal strain of Penicillium oxalicum TWSF1 and solid fermenting and culturing it to obtain a fungal culture; Step 4: washing the fungal culture with a Tween-80 solution, filtering the washing solution obtained by washing to obtain a spore suspension, centrifuging the spore suspension, and then precipitating and freeze-drying to obtain fungal spore powder; Step 5, testing the effective viable count of the two initial bacterial inoculants and fungal spore powder; Step 6: According to the total effective live bacteria count of the composite bacterial agent, two initial bacterial agents and fungal spore powder are added in proportion, and the remaining part is filled with an adsorbent, and mixed evenly to obtain the composite bacterial agent.

[0021] The composite bacterial agent can be processed into tablets, granules or powders, and applied to the planting of crops in slightly saline-alkali lands in cold northern regions, and can also be applied to nitrogen fixation, phosphorus solubilization and salinization improvement of soils.

[0022] Example 3

[0023] The composite bacterial agent for saline-alkali land of the present invention comprises Pseudomonas mohnii TWS2N1, Pseudomonas rustica TWS23 and Penicillium oxalicum TWSF1, and the effective live bacteria count ratio is 1:2:2, and the total effective live bacteria count of the composite bacterial agent is not less than 2.0×10 8 CFU / g.

[0024] Among them, the Pseudomonas mohnii TWS2N1 has the function of fixing nitrogen and producing the plant growth hormone indoleacetic acid (IAA), the Pseudomonas rustica TWS23 has the function of dissolving inorganic phosphorus and producing the plant growth hormone indoleacetic acid (IAA), and the Penicillium oxalicum TWSF1 is salt-alkali resistant and can dissolve inorganic phosphorus.

[0025] A method for preparing a composite bacterial agent for saline-alkali land comprises the following steps: Step 1, activating and fermenting the bacterial strains of Pseudomonas mohnii TWS2N1 and Pseudomonas rustica TWS23 respectively, and centrifuging to obtain bacterial sludge of the two bacteria; Step 2: mixing the bacterial sludge of the two bacteria with the adsorbent respectively, and freeze-drying them to obtain two initial bacterial agents; Step 3, activating the fungal strain of Penicillium oxalicum TWSF1 and solid fermenting and culturing it to obtain a fungal culture; Step 4: washing the fungal culture with a Tween-80 solution, filtering the washing solution obtained by washing to obtain a spore suspension, centrifuging the spore suspension, and then precipitating and freeze-drying to obtain fungal spore powder; Step 5, testing the effective viable count of the two initial bacterial inoculants and fungal spore powder; Step 6: According to the total effective live bacteria count of the composite bacterial agent, two initial bacterial agents and fungal spore powder are added in proportion, and the remaining part is filled with an adsorbent, and mixed evenly to obtain the composite bacterial agent.

[0026] The composite bacterial agent can be processed into tablets, granules or powders, and applied to the planting of crops in slightly saline-alkali lands in cold northern regions, and can also be applied to nitrogen fixation, phosphorus solubilization and salinization improvement of soils.

[0027] Example 4

[0028] A method for preparing a composite bacterial agent for saline-alkali land comprises the following steps: Step 1, activating and fermenting the bacterial strains of Pseudomonas mohnii TWS2N1 and Pseudomonas rustica TWS23 respectively, and centrifuging to obtain bacterial sludge of the two bacteria; Step 2: mixing the bacterial sludge of the two bacteria with the adsorbent respectively, and freeze-drying them to obtain two initial bacterial agents; Step 3, activating the fungal strain of Penicillium oxalicum TWSF1 and solid fermenting and culturing it to obtain a fungal culture; Step 4: washing the fungal culture with a Tween-80 solution, filtering the washing solution obtained by washing to obtain a spore suspension, centrifuging the spore suspension, and then precipitating and freeze-drying to obtain fungal spore powder; Step 5: The effective viable count of the two initial bacterial agents and fungal spore powder was tested. The test showed that the effective viable count of the initial bacterial agent of Pseudomonas TWS2N1 was 3.5×10 8 CFU / g, the effective viable count of the initial bacterial inoculum of Pseudomonas TWS23 was 4.8×10 8 CFU / g, the number of effective viable bacteria in the fungal spore powder of Penicillium oxalicum TWSF1 was 10.7×10 8 CFU / g; Step 6: The total effective viable count is 2.0×10 8CFU / g, according to the ratio of effective live bacteria count of Pseudomonas mohnii TWS2N1: Pseudomonas rustica TWS23: Penicillium oxalicum TWSF = 2:2:1, two initial bacterial agents and fungal spore powder are added, and the rest is filled with adsorbent, and mixed evenly to obtain a composite agent.

[0029] Among them, the Pseudomonas mohnii TWS2N1 has the function of fixing nitrogen and producing the plant growth hormone indoleacetic acid (IAA), the Pseudomonas rustica TWS23 has the function of dissolving inorganic phosphorus and producing the plant growth hormone indoleacetic acid (IAA), and the Penicillium oxalicum TWSF1 is salt-alkali resistant and can dissolve inorganic phosphorus.

[0030] The composite bacterial agent can be processed into tablets, granules or powders, and applied to the planting of crops in slightly saline-alkali lands in cold northern regions, and can also be applied to nitrogen fixation, phosphorus solubilization and salinization improvement of soils.

[0031] Taking wheat as an example, the growth-promoting effect of compound fungicides on crops was explored.

[0032] In the turnover box with the inner size of 615mm×372mm×149mm, add 30kg of light saline-alkali soil, add appropriate amount of water to keep the humidity of each pot soil consistent, and control the amount of watering for each pot to be the same each time. The compound bacterial agent is applied with water, and 2g of compound bacterial agent is diluted 1000 times for each pot before application.

[0033] The potted plants not inoculated with the composite microbial agent were used as the blank control group CK, the potted plants inoculated with other commercially available multifunctional microbial agents were used as T1, and the potted plants inoculated with the composite microbial agent provided by the present invention were used as T2. Each treatment was repeated 5 times.

[0034] When the outdoor temperature was below 20℃, after 30 days of outdoor cultivation, all wheat plants were pulled out, and the rhizosphere soil was collected to measure the plant height, leaf length, root length, and fresh weight of the wheat; the pH, available nitrogen, and available phosphorus content of the rhizosphere soil were determined. The results are shown in Tables 1 and 2. Table 1 Effects of different treatments on wheat growth deal with Plant height / cm Leaf length / cm Root length / cm Fresh weight / g CK 25.6±1.2 8.4±0.3 9.1±0.1 2.1±0.2 T1 26.7±1.4 9.3±0.2 10.2±0.4 2.4±0.1 T2 29.5±1.5 12.6±0.3 12.7±0.3 3.2±0.1 Table 2 Effects of different treatments on soil

[0035] The experimental results are shown in Tables 1 and 2. Under low temperature conditions, the application of the composite microbial agent provided by the present invention when planting wheat in saline-alkali land can effectively promote the increase of wheat plant height, leaf length, root length, and fresh weight, reduce soil pH, and increase the content of soil available nutrients, and is superior to other multifunctional microbial agents available on the market.

[0036] The parts not described in detail herein are prior art. Experimental methods without specific conditions in the examples are usually carried out under conventional conditions or under conditions recommended by the manufacturer, and experimental steps not described in detail are referred to the prior art database.

[0037] Although the specific embodiments of the present invention are described in detail above, the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in this field without departing from the purpose of the present invention, and modifications or deformations without creative labor are still within the protection scope of the present invention.

Claims

1. A composite bacterial agent for saline-alkali land, characterized in that: Including Pseudomonas mohnii TWS2N1, Pseudomonas rustica TWS23 and Penicillium oxalicum TWSF1, the Pseudomonas mohnii TWS2N1 has been deposited in the General Microbiology Center of China National Microbiological Culture Collection on October 25, 2024, with a deposit number of CGMCC No.32348, the Pseudomonas rustica TWS23 has been deposited in the General Microbiology Center of China National Microbiological Culture Collection on September 25, 2024, with a deposit number of CGMCC No.32078, and the Penicillium oxalicum TWSF1 has been deposited in the General Microbiology Center of China National Microbiological Culture Collection on October 25, 2024, with a deposit number of CGMCC No.41553.

2. The composite bacterial agent for saline-alkali land according to claim 1, characterized in that: The ratio of the effective viable bacteria counts of Pseudomonas mohnii TWS2N1, Pseudomonas rustica TWS23 and Penicillium oxalicum TWSF1 is 1-2:1-2:1-2, and the total effective viable bacteria count of the composite bacterial agent is not less than 2.0×10 8 CFU / g.

3. The composite bacterial agent for saline-alkali land according to claim 2, characterized in that: The composite bacterial agent is in the form of tablets, granules or powders.

4. The composite bacterial agent for saline-alkali land according to claim 3, characterized in that: The preparation method comprises the following steps: Step 1, activating and fermenting the bacterial strains of Pseudomonas mohnii TWS2N1 and Pseudomonas rustica TWS23 respectively, and centrifuging to obtain bacterial sludge of the two bacteria; Step 2: mixing the bacterial sludge of the two bacteria with the adsorbent respectively, and freeze-drying them to obtain two initial bacterial agents; Step 3, activating the fungal strain of Penicillium oxalicum TWSF1 and solid fermenting and culturing it to obtain a fungal culture; Step 4: washing the fungal culture with a Tween-80 solution, filtering the washing solution obtained by washing to obtain a spore suspension, centrifuging the spore suspension, and then precipitating and freeze-drying to obtain fungal spore powder; Step 5, testing the effective viable count of the two initial bacterial inoculants and fungal spore powder; Step 6: According to the total effective live bacteria count of the composite bacterial agent, two initial bacterial agents and fungal spore powder are added in proportion, and the remaining part is filled with an adsorbent, and mixed evenly to obtain the composite bacterial agent.

5. Use of the composite bacterial agent for saline-alkali land according to any one of claims 1 to 4 in the cultivation of crops in slightly saline-alkali land in northern cold regions.

6. Use of the composite bacterial agent for saline-alkali land according to any one of claims 1 to 4 in soil nitrogen fixation, phosphorus solubilization and improvement of salinization.

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