Application of compound microbial agent to promotion of phosphorus and potassium solubilization of soil and improvement of available phosphorus and potassium content of soil

Abstract

The invention relates to an application of a compound microbial agent in promoting phosphorus and potassium solubilization of soil and increasing the content of available phosphorus and potassium in the soil. The bacterial strains in the compound microbial agent consist of a bacillus coagulans BC66 bacterial strain, a lactobacillus plantarum LP194 bacterial strain and a lactobacillus acidophilus LA71 bacterial strain. The invention also relates to an application of the compound microbial agent in promoting phosphorus and potassium solubilization of the soil and increasing the content of available phosphorus and potassium in the soil, and the bacterial strains in the compound microbial agent consist of a bacillus coagulans BC66 bacterial strain, a lactobacillus plantarum LP194 bacterial strain and a lactobacillus acidophilus LA71 bacterial strain. The invention develops a brand-new microbial preparation capable of promoting the phosphorus and potassium solubilizing effect of soil, and researches find that the three strains are mutually matched and have synergistic interaction on the effects of promoting phosphorus and potassium solubilizing of soil, improving the content of available phosphorus and potassium in soil, improving the utilization rate of phosphate fertilizer and potassium fertilizer and increasing the yield of fruit trees, and the yield of the fruit trees is increased under the condition that the using bacterium amount is consistent. Compared with a single strain intervention mode or other strain combined intervention mode, the microbial preparation has the advantage that the effects are obviously improved. Therefore, the compound microbial agent provides a new strategy for promoting phosphorus and potassium solubilization of soil, increasing the content of available phosphorus and potassium in soil, increasing the utilization rate of phosphate fertilizer and potassium fertilizer and increasing the yield of fruit trees.

Description

Technical Field

[0001] The invention belongs to the technical field of soil fertility improvement, and relates to the application of a composite microbial agent in promoting soil phosphorus and potassium dissolution and increasing the effective phosphorus and potassium content in the soil. Background Art

[0002] In order to ensure food security, reduce the negative impact of excessive use of fertilizers on the environment, improve soil fertility, and develop green agriculture, it has become an inevitable trend. Although fertilizers play an important role in modern agriculture, the utilization efficiency of fertilizers has been low for a long time due to improper fertilization and soil condition restrictions. Among them, the utilization rate of phosphate fertilizers and potassium fertilizers is particularly low. A large amount of phosphorus and potassium elements are fixed or lost in the soil and cannot be effectively used, resulting in soil acidification and compaction, reducing crop yields and quality.

[0003] As a new type of biological fertilizer, microbial agents have many advantages. First, microbial agents can use their own metabolic activities to decompose insoluble nutrients in the soil, such as fixed phosphorus and potassium, and convert them into effective nutrients that can be absorbed and utilized by crops. Secondly, microbial agents can improve soil structure, enhance soil water and fertilizer retention capacity, and improve soil fertility. In addition, microbial agents also have multiple functions such as disease resistance and insect suppression, and promote crop growth, which can significantly increase the yield and quality of crops.

[0004] However, the existing phosphorus-potassium-solubilizing microbial agents still have some limitations in practical applications, such as unstable strain activity and single formula, which lead to insignificant phosphorus-potassium-solubilizing effects. Therefore, it is very meaningful to develop a new type of phosphorus-potassium-solubilizing microbial agent to effectively increase the effective phosphorus and potassium content in the soil. Summary of the invention

[0005] In view of the shortcomings of the prior art, the present invention aims to provide an application of a composite microbial agent in promoting soil phosphorus and potassium dissolution and increasing the effective phosphorus and potassium content in the soil. The composite microbial agent can promote soil nutrient conversion and increase fruit tree yield.

[0006] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides an application of a composite microbial agent in promoting soil phosphorus and potassium solubilization and increasing the effective phosphorus and potassium content in the soil, characterized in that the strains in the composite microbial agent are composed of Bacillus coagulans BC66 strain with a preservation number of CGMCC 21879, Lactobacillus plantarum LP194 strain with a preservation number of CGMCC 21766, and Lactobacillus acidophilus LA71 strain with a preservation number of CGMCC 21765.

[0008] The present invention has developed a new microbial preparation for promoting the effect of soil phosphorus and potassium dissolution, which is a combination of three strains of Bacillus coagulans BC66, Lactobacillus plantarum LP194, and Lactobacillus acidophilus LA71. Studies have found that these three strains cooperate with each other in promoting soil phosphorus and potassium dissolution, increasing the effective phosphorus and potassium content in the soil, and increasing the utilization rate of phosphate fertilizer and potassium fertilizer and the yield of fruit trees. When the amount of bacteria is consistent, compared with a single strain intervention method or other strain combined intervention methods, the microbial preparation is significantly improved in the above-mentioned effects. Therefore, the composite microbial agent provides a new strategy for promoting soil phosphorus and potassium dissolution, increasing the effective phosphorus and potassium content in the soil, and increasing the utilization rate of phosphate fertilizer and potassium fertilizer and the yield of fruit trees.

[0009] Preferably, the ratio of the viable cell counts of the BC66 strain, the LP194 strain and the LA71 strain is (1-8):(2-5):(2-5).

[0010] Among them, the specific point values ​​in (1-8) can be selected from 1, 2, 3, 4, 5, 6, 7, 8, etc.; the specific point values ​​in (2-5) can be selected from 2, 3, 4, 5, etc.; other specific point values ​​within the above numerical range can be selected, and they will not be listed here one by one.

[0011] Based on the potential interaction relationship between Bacillus coagulans BC66, Lactobacillus plantarum LP194 and Lactobacillus acidophilus LA71, the present invention also found that when the three strains are compounded in the above-mentioned specific ratio of live bacteria count, their effects in promoting soil phosphorus and potassium solubilization, increasing the effective phosphorus and potassium content in the soil, improving the utilization rate of phosphorus and potassium fertilizers and fruit tree yield are more significant.

[0012] Preferably, the content of live bacteria in the composite microbial agent is not less than 5×10 9 CFU / g or 5×10 9 CFU / mL, for example 5×10 9 CFU / mL (CFU / g), 1×10 10CFU / mL (CFU / g), 1×10 11 CFU / mL (CFU / g), 1×10 12 CFU / mL (CFU / g), etc.; other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0013] Preferably, the composite microbial agent is in the form of a solution or a powder.

[0014] The dosage form of the composite microbial agent of the present invention is not limited, including the most commonly used solutions, powders, or further prepared capsules, tablets or granules.

[0015] Preferably, when the composite microbial agent is in the form of a powder, it is prepared by a method comprising the following steps:

[0016] The BC66 strain, the LP194 strain and the LA71 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged respectively, mixed with a protective agent and then dried to obtain BC66 bacterial powder, LP194 bacterial powder and LA71 bacterial powder; the BC66 bacterial powder, LP194 bacterial powder and LA71 bacterial powder are mixed according to the ratio of the number of live bacteria to obtain the composite microbial inoculant.

[0017] Alternatively, the BC66 strain, the LP194 strain and the LA71 strain are inoculated into a culture medium together for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged, mixed with a protective agent and then dried to obtain the composite microbial agent.

[0018] The protective agent includes any one of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinyl pyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol, or a combination of at least two thereof.

[0019] Preferably, when the composite microbial agent is in the form of a solution, it is prepared by a method comprising the following steps:

[0020] The BC66 strain, the LP194 strain and the LA71 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged respectively, and the bacterial mud is resuspended to obtain a BC66 bacterial suspension, a LP194 bacterial suspension and a LA71 bacterial suspension; the BC66 bacterial suspension, the LP194 bacterial suspension and the LA71 bacterial suspension are mixed according to the ratio of the number of viable bacteria to obtain the composite microbial inoculant.

[0021] Alternatively, the BC66 strain, the LP194 strain and the LA71 strain are inoculated into a culture medium together for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged, and the bacterial sludge is resuspended to obtain the composite microbial agent.

[0022] Preferably, the composite microbial agent also contains a bacterial protective agent and / or a nutritional component.

[0023] In a second aspect, the present invention provides a method for promoting soil phosphorus and potassium dissolution and increasing fruit tree yield, the method comprising: mixing a composite microbial agent with water and applying the mixture to fruit tree planting soil;

[0024] The strains in the composite microbial agent are composed of Bacillus coagulans BC66 strain with a preservation number of CGMCC 21879, Lactobacillus plantarum LP194 strain with a preservation number of CGMCC 21766, and Lactobacillus acidophilus LA71 strain with a preservation number of CGMCC 21765.

[0025] The present invention also develops a new strategy for increasing fruit tree yields, that is, a composite microbial agent comprising BC66 strain, LP194 strain and LA71 strain is mixed with water and applied to fruit tree planting soil, and the three strains cooperate with each other in improving the utilization rate of phosphate fertilizer and potassium fertilizer and fruit tree yield, and synergistically increase the effect. The agent can effectively increase the effective phosphorus and potassium content in the soil and the yield of fruit trees by applying efficient phosphorus and potassium solubilizing strains and combining the growth characteristics of fruit trees, and improve the absorption and utilization of soil nutrients by fruit trees. The development of this technology is of great significance for achieving sustainable development of agriculture.

[0026] Preferably, the ratio of the viable cell counts of the BC66 strain, the LP194 strain and the LA71 strain is (1-8):(2-5):(2-5).

[0027] Among them, the specific point values ​​in (1-8) can be selected from 1, 2, 3, 4, 5, 6, 7, 8, etc.; the specific point values ​​in (2-5) can be selected from 2, 3, 4, 5, etc.; other specific point values ​​within the above numerical range can be selected, and they will not be listed here one by one.

[0028] Preferably, the composite microbial agent is in the form of a solution or a powder.

[0029] Preferably, the content of live bacteria in the composite microbial agent is not less than 5×10 9 CFU / g or 5×10 9 CFU / mL, for example 5×10 9 CFU / mL (CFU / g), 1×10 10 CFU / mL (CFU / g), 1×10 11 CFU / mL (CFU / g), 1×10 12CFU / mL (CFU / g), etc.; other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0030] Preferably, the usage of the composite microbial agent is 10-30kg / mu, for example, 10kg / mu, 12kg / mu, 15kg / mu, 18kg / mu, 20kg / mu, 22kg / mu, 25kg / mu, 30kg / mu, etc.; other specific point values ​​within this numerical range can be selected, and they will not be described one by one here.

[0031] Preferably, the mode of application includes flushing.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] The present invention has developed a new microbial preparation for promoting the effect of soil phosphorus and potassium dissolution, which is a combination of three strains of Bacillus coagulans BC66, Lactobacillus plantarum LP194, and Lactobacillus acidophilus LA71. Studies have found that these three strains cooperate with each other in promoting soil phosphorus and potassium dissolution, increasing the effective phosphorus and potassium content in the soil, and increasing the utilization rate of phosphate fertilizer and potassium fertilizer and the yield of fruit trees. When the amount of bacteria is consistent, compared with a single strain intervention method or other strain combined intervention methods, the microbial preparation is significantly improved in the above-mentioned effects. Therefore, the composite microbial agent provides a new strategy for promoting soil phosphorus and potassium dissolution, increasing the effective phosphorus and potassium content in the soil, and increasing the utilization rate of phosphate fertilizer and potassium fertilizer and the yield of fruit trees.

[0034] The BC66 strain involved in the present invention is classified and named Bacillus coagulans. The preservation unit is the General Microbiology Center of China Microorganism Culture Collection Administration. The preservation time is March 8, 2021. The preservation number is CGMCC 21879. The address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0035] The LP194 strain involved in the present invention is classified and named Lactobacillus plantarum, and the preservation unit is the General Microbiology Center of China Microorganism Culture Collection Administration. The preservation time is January 29, 2021, the preservation number is CGMCC 21766, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0036] The LA71 strain involved in the present invention is classified and named Lactobacillus acidophilus, and the preservation unit is the General Microbiology Center of China Microorganism Culture Collection Administration. The preservation time is January 29, 2021, the preservation number is CGMCC 21765, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. DETAILED DESCRIPTION

[0037] In order to further explain the technical means and effects adopted by the present invention, the technical solution of the present invention is further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0038] The BC66 strain involved in the following experiment is classified and named Bacillus coagulans, deposited on March 8, 2021, and has a deposit number of CGMCC 21879.

[0039] The LP194 strain involved in the following experiment is classified and named Lactobacillus plantarum, the preservation date is January 29, 2021, and the preservation number is CGMCC 21766.

[0040] The LA71 strain involved in the following experiment is classified and named Lactobacillus acidophilus, the preservation date is January 29, 2021, and the preservation number is CGMCC 21765.

[0041] The preparation method of the bacterial solution involved in the following test is as follows: the desired strain is inoculated into MRS liquid culture medium, cultured at 37°C for 24 hours for activation, and activated twice continuously to obtain an activated solution; the activated solution is inoculated into MRS liquid culture medium at an inoculum volume of 5% (v / v), and cultured at 37°C to obtain a bacterial solution; the number of viable bacteria in each bacterial solution is 2×10 10 CFU / mL.

[0042] Example 1

[0043] Verification of the effect of composite microbial agents on soil phosphorus and potassium dissolution:

[0044] (1) Experimental groups: 13 groups in total:

[0045] Blank control group;

[0046] Medium-dose intervention group: BC66+LP194 group (intervention with BC66 bacterial solution and LP194 bacterial solution, the ratio of live bacteria count was 5:3),

[0047] LP194+LA71 group (intervention with LP194 bacterial solution and LA71 bacterial solution, the ratio of viable bacteria count was 3:2), BC66+LA71 group (intervention with BC66 bacterial solution and LA71 bacterial solution, the ratio of viable bacteria count was 5:2);

[0048] BC66+LP194+LA71 group 1 (combined intervention of three bacterial solutions, the ratio of live bacteria count was 5:3:2),

[0049] BC66+LP194+LA71 group 2 (combined intervention with three bacterial solutions, the ratio of live bacteria count was 2:3:5),

[0050] BC66+LP194+LA71 group 3 (combined intervention of three bacterial solutions, the ratio of live bacteria count was 7:2:3),

[0051] ATCC7050+ATCC8014+ATCC4356 group (commercially available Bacillus coagulans liquid, commercially available Lactobacillus plantarum liquid and Lactobacillus acidophilus liquid were used for intervention, with the ratio of viable bacteria count being 5:3:2);

[0052] BC66 group (intervention with BC66 bacterial solution),

[0053] LP194 group (intervention with LP194 bacterial solution),

[0054] LA71 group (intervention with LA71 bacterial solution);

[0055] Low-dose intervention group: BC66+LP194+LA71 group 1 (combined intervention with three bacterial solutions, the ratio of live bacteria count was 5:3:2);

[0056] High-dose intervention group: BC66+LP194+LA71 group 1 (combined intervention of three bacterial liquids, the ratio of live bacteria count was 5:3:2).

[0057] (2) Experimental process: In the potted plant experiment, the soil from which apples were planted was taken out and transferred to each pot. Each pot was filled with 5 kg of soil. The bacterial solution or mixed bacterial solution of each group was first mixed with water and then flushed to the soil. The blank control group used water without bacterial solution (600 g) for flushing; the medium-dose intervention group used water containing 0.6 mL of bacterial solution or 0.6 mL of mixed bacterial solution (600 g) for flushing; the low-dose intervention group used water containing 0.3 mL of mixed bacterial solution (600 g) for flushing; and the high-dose intervention group used water containing 1.2 mL of mixed bacterial solution (600 g) for flushing.

[0058] (3)Indicator analysis:

[0059] The available phosphorus and available potassium content of each group of soil were tested on the 1st, 30th, 90th and 180th day of the experiment. The determination method of available phosphorus and available potassium content of soil was referred to "Soil Agrochemical Analysis" (Bao Shidan, third edition). The results are shown in Table 1.

[0060] Table 1

[0061]

[0062]

[0063]

[0064] It can be seen from the data results in Table 1 that, compared with the blank control group or commercially available microbial agents, the composite microbial agent involved in the present invention can effectively promote soil phosphorus and potassium solubilization and increase the effective phosphorus and potassium content in the soil, and the three strains of Bacillus coagulans BC66, Lactobacillus plantarum LP194 and Lactobacillus acidophilus LA71 have a synergistic effect on the above effects.

[0065] Example 2

[0066] Verification of the yield-increasing effect of compound microbial agents on fruit trees:

[0067] (1) Experimental groups: 10 groups in total:

[0068] Blank control group;

[0069] Medium-dose intervention group: BC66+LP194 group (intervention with BC66 bacterial solution and LP194 bacterial solution, the ratio of live bacteria count was 5:3),

[0070] LP194+LA71 group (intervention with LP194 bacterial solution and LA71 bacterial solution, the ratio of live bacteria count was 3:2),

[0071] BC66+LA71 group (intervention with BC66 bacterial solution and LA71 bacterial solution, the ratio of viable bacteria count was 5:2);

[0072] BC66+LP194+LA71 group 1 (combined intervention of three bacterial solutions, the ratio of live bacteria count was 5:3:2),

[0073] BC66+LP194+LA71 group 2 (combined intervention with three bacterial solutions, the ratio of live bacteria count was 2:3:5),

[0074] BC66+LP194+LA71 group 3 (combined intervention of three bacterial solutions, the ratio of live bacteria count was 7:2:3),

[0075] ATCC7050+ATCC8014+ATCC4356 group (commercially available Bacillus coagulans liquid, commercially available Lactobacillus plantarum liquid and Lactobacillus acidophilus liquid were used for intervention, with the ratio of viable bacteria count being 5:3:2);

[0076] Low-dose intervention group: BC66+LP194+LA71 group 1 (combined intervention with three bacterial solutions, the ratio of live bacteria count was 5:3:2);

[0077] High-dose intervention group: BC66+LP194+LA71 group 1 (combined intervention of three bacterial liquids, the ratio of live bacteria count was 5:3:2).

[0078] (2) Experimental process: Field orchard tests were conducted at an apple planting base in Yantai City. The experimental plots were divided into ten plots on average, and the cultivation and management measures of each group were the same. The bacterial solution or mixed bacterial solution of each group was first mixed with water and then flushed into the planting soil. The blank control group used water without bacterial solution (20 tons / mu) for flushing; the medium-dose intervention group used water containing bacterial solution or mixed bacterial solution (20 kg / mu) for flushing; the low-dose intervention group used water containing bacterial solution or mixed bacterial solution (10 kg / mu) for flushing; the high-dose intervention group used water containing bacterial solution or mixed bacterial solution (30 kg / mu) for flushing. Each group was flushed three times (March 31, May 31, and July 31), with an interval of 2 months each time.

[0079] (3)Indicator analysis:

[0080] The soil available phosphorus and quick-acting potassium contents were tested on the 30th day after the three irrigations and at the maturity stage, and the results are shown in Table 2. The actual yield was measured at the apple maturity stage in late October, and the results are shown in Table 3.

[0081] Table 2

[0082]

[0083]

[0084]

[0085] Table 3

[0086]

[0087] From the data results in Table 2 and Table 3, it can be seen that compared with the blank control group or the commercially available microbial agent, the composite microbial agent involved in the present invention can effectively increase the soil available phosphorus and potassium content and apple yield during apple planting, indicating that the composite microbial agent of the present invention continuously promotes the release of soil phosphorus and potassium during the apple growth period, increases the soil available phosphorus and potassium content, thereby increasing apples' absorption of soil phosphorus and potassium, and increasing apple yield. Moreover, the three strains of Bacillus coagulans BC66, Lactobacillus plantarum LP194, and Lactobacillus acidophilus LA71 have a synergistic effect on the above effects.

[0088] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0089] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0090] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. An application of a composite microbial agent in promoting soil phosphorus and potassium dissolution and increasing the effective phosphorus and potassium content in the soil, characterized in that: The strains in the composite microbial agent are composed of Bacillus coagulans BC66 strain with a preservation number of CGMCC 21879, Lactobacillus plantarum LP194 strain with a preservation number of CGMCC 21766, and Lactobacillus acidophilus LA71 strain with a preservation number of CGMCC 21765.

2. The use according to claim 1, characterized in that: The ratio of the number of viable bacteria among the BC66 strain, LP194 strain and LA71 strain is (1-8):(2-5):(2-5).

3. The use according to claim 1, characterized in that: The content of live bacteria in the composite microbial agent is not less than 5×10 9 CFU / g or 5×10 9 CFU / mL.

4. The use according to claim 1, characterized in that: The dosage form of the composite microbial agent includes solution or powder.

5. The use according to claim 1, characterized in that: The composite microbial agent also contains a bacterial protective agent and / or a nutritional component.

6. A method for promoting soil phosphorus and potassium dissolution and increasing fruit tree yield, characterized in that: The method comprises: mixing the composite microbial agent with water and applying the mixture to the fruit tree planting soil; The strains in the composite microbial agent are composed of Bacillus coagulans BC66 strain with a preservation number of CGMCC 21879, Lactobacillus plantarum LP194 strain with a preservation number of CGMCC 21766, and Lactobacillus acidophilus LA71 strain with a preservation number of CGMCC 21765.

7. The method according to claim 6, characterized in that The ratio of the number of viable bacteria among the BC66 strain, LP194 strain and LA71 strain is (1-8):(2-5):(2-5).

8. The method according to claim 6, characterized in that The dosage form of the composite microbial agent is a solution or a powder; the content of live bacteria in the composite microbial agent is not less than 5×10 9 CFU / g or 5×10 9 CFU / mL.

9. The method according to claim 8, characterized in that The usage of the composite microbial agent is 10-30 kg / mu.

10. The method according to claim 6, characterized in that The application method includes flushing.

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