A compound microbial agent for improving corn yield, a preparation method and application thereof
By preparing a compound microbial agent containing Enterobacteriaceae and Pseudomonas, the problems of soil degradation and yield reduction in maize cultivation were solved, achieving the effects of soil improvement and increased maize yield.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
- Filing Date
- 2026-02-25
- Publication Date
- 2026-06-02
AI Technical Summary
Current corn cultivation faces problems such as soil degradation, poor crop development and reduced yield, and increased pests and diseases. Furthermore, excessive use of pesticides and fertilizers leads to environmental pollution and increased planting costs.
A compound microbial agent containing Enterobacter sp. X1 and Pseudomonas sp. X2, along with wood ash, vermiculite, sodium carboxymethyl cellulose, trehalose, and humic acid, was prepared to improve soil physicochemical properties and promote corn growth.
It significantly increases soil organic carbon content, improves soil physical and chemical properties, promotes corn growth, increases yield and income, and achieves a significant increase in corn production.
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Figure CN122127172A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the fields of agriculture and biotechnology, specifically to a compound microbial agent for improving corn production capacity, its preparation method, and its application. Background Technology
[0002] As a staple food crop, corn plays a crucial role in ensuring food production. To alleviate the contradiction between limited land resources and a growing population, corn production systems are gradually trending towards highly intensive cultivation. Currently, corn cultivation faces many problems, mainly manifested in soil degradation, poor crop development and reduced yields, and increased pests and diseases. Furthermore, excessive use of pesticides and fertilizers not only exacerbates soil acidification and compaction but also leads to a decline in corn quality, increased environmental pollution, and higher planting costs.
[0003] Compound microbial inoculants are formulated by mixing and cultivating several microorganisms with different functional groups and symbiotic or mutualistic relationships, resulting in an agent with stronger growth-promoting capabilities and greater stability. Using compound microbial inoculants to replace traditional chemical fertilizers can effectively improve crop growth and reduce environmental pollution. Therefore, developing a stable and efficient compound microbial inoculant that can be applied in the field and significantly improve crop yield is of significant practical importance. Summary of the Invention
[0004] The purpose of this invention is to provide a compound microbial agent for improving corn production capacity, its preparation method and application. When applied to farmland, this compound microbial agent can improve the physical and chemical properties of the soil and increase corn yield.
[0005] This invention relates to a compound microbial agent for improving corn production capacity, comprising a compound bacterial solution and a compound carrier. The compound bacterial solution includes an Enterobacter suspension and a Pseudomonas suspension, and the compound carrier includes wood ash, vermiculite, sodium carboxymethyl cellulose, trehalose, and humic acid.
[0006] The Enterobacter sp. X1 was deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, on January 6, 2026, with accession number CCTCC NO: M 2026023.
[0007] The Pseudomonas species described is Pseudomonas sp. X2, deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, on January 6, 2026, with accession number CCTCC NO: M 2026024.
[0008] This invention provides a method for preparing the above-mentioned composite microbial agent, comprising the following steps:
[0009] 1. Enterobacteriaceae and Pseudomonas were streaked onto TSB solid medium to obtain two activated strains.
[0010] 2. Using an inoculation needle, two activated bacterial strains were picked up and inoculated separately into TSB liquid medium to obtain Enterobacter suspension and Pseudomonas suspension; the OD values of the two bacterial suspensions were measured. 600 The value is 0.8~1.0;
[0011] 3. Dry the wood ash and sterilize it to obtain treated wood ash; soak the vermiculite in CaCl2 solution, dry it until the moisture content is <5%, sterilize it, and obtain treated vermiculite;
[0012] 4. Mix the treated wood ash from step 3 with the treated vermiculite at a volume ratio of 1:1 to obtain a mixed matrix; add sodium carboxymethyl cellulose, trehalose, and humic acid to the mixed matrix to obtain a compound carrier;
[0013] 5. Mix the Enterobacter suspension and Pseudomonas suspension to form a compound bacterial solution; mix the compound bacterial solution with the compound carrier at a volume ratio of 1:1, air dry, and then mature at room temperature. Finally, adjust the moisture content to 25% to obtain the compound microbial agent.
[0014] Furthermore, in step three, the plant ash has a pH of 7.0 and an electrical conductivity of <1 ms / cm.
[0015] Furthermore, in step three, the wood ash is sterilized by high-pressure steam sterilization at 121℃ for 30-40 minutes.
[0016] Furthermore, in step three, the concentration of the CaCl2 solution is 0.05~0.15 mol / L.
[0017] Furthermore, the soaking time in step three is 2-3 hours.
[0018] Furthermore, in step three, the vermiculite is sterilized by dry heat at 160~170℃ for 2~3 hours.
[0019] This invention provides the application of the above-mentioned compound microbial agent in soil improvement.
[0020] This invention provides the application of the above-mentioned compound microbial agent in promoting corn growth.
[0021] This invention provides the application of the above-mentioned compound microbial agent in increasing corn yield.
[0022] The beneficial effects of this invention are:
[0023] The active ingredients in the compound microbial agent of this invention are Enterobacter sp. X1 and Pseudomonas sp. X2. The compound microbial agent prepared by this invention has phosphorus-solubilizing and nitrogen-fixing capabilities, and can also effectively increase the organic carbon content in the soil and improve the soil's physical and chemical properties. The compound microbial agent of this invention can promote corn growth, achieving the goal of increasing yield and income. Attached Figure Description
[0024] Figure 1 Phylogenetic tree of Enterobacter X1;
[0025] Figure 2 Phylogenetic tree of Pseudomonas X2;
[0026] Figure 3 Photograph of the composite microbial inoculant prepared in Example 3;
[0027] Figure 4 The change in soil pH after the application of compound microbial inoculant;
[0028] Figure 5 The change in soil EC value after application of compound microbial inoculant;
[0029] Figure 6 Changes in soil SOC after application of compound microbial inoculant;
[0030] Figure 7 Changes in soil total nitrogen (TN) after application of compound microbial inoculant;
[0031] Figure 8 Changes in soil total phosphorus (TP) after application of compound microbial inoculant;
[0032] Figure 9 The effect of compound microbial inoculants on promoting the fresh weight of corn plants;
[0033] Figure 10 The effect of compound microbial inoculants on promoting the dry weight of corn plants;
[0034] Figure 11 The effect of compound microbial inoculants on promoting the height of corn plants;
[0035] Figure 12 The effect of compound microbial inoculants on corn yield. Detailed Implementation
[0036] The embodiments of the present invention will be described in detail below. The following embodiments are implemented based on the technical solution of the present invention, and detailed implementation schemes and specific operation processes are given. However, the protection scope of the present invention is not limited to the following embodiments.
[0037] Example 1: Isolation, screening and identification of Enterobacteriaceae
[0038] The Enterobacter sp. X1 in this embodiment is deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, China, on January 6, 2026, with accession number CCTCC NO: M 2026023.
[0039] 1. Isolation and purification of Enterobacter X1
[0040] Fresh rhizosphere soil from corn was taken, and under aseptic conditions, it was added to sterile water, shaken, diluted, and spread on 50% TSB solid agar plates. After multiple isolation and purification processes, a single colony was obtained, which was Enterobacter X1.
[0041] 2. Identification of Enterobacter X1
[0042] Enterobacter X1 colonies were cultured on TSB solid medium for 48 hours. The colonies were grayish-white, oval, opaque, with neat edges, and Gram-negative.
[0043] Using Enterobacter X1 DNA as a template, PCR amplification was performed using universal 16S rDNA primers 27F and 1492R. Primer 27F was 5'-AGAGTTTGATCCTGGCTCAG-3'; primer 1492R was 5'-GGTTACCTTGTTACGACTT-3'. Sequencing was performed after amplification, and the 16S rDNA sequence is shown in SEQ ID NO: 1 of the sequence listing. The obtained Enterobacter X1 16S rDNA sequencing results were entered into the NCBI database for BLAST alignment to obtain the relevant type strain. After preliminary processing of the target sequence and type strain sequence using MEGA11, a phylogenetic tree was constructed according to the Neighbor-joining method. The results are shown below. Figure 1 As shown, Enterobacter X1 is highly similar to Enterobacter sp. in its 16S rRNA gene, thus identifying Enterobacter X1 as Enterobacter sp., belonging to the genus Enterobacter.
[0044] Example 2: Isolation, screening and identification of Pseudomonas aeruginosa
[0045] The Pseudomonas strain in this embodiment is Pseudomonas sp. X2, deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, China, on January 6, 2026, with accession number CCTCC NO: M 2026024.
[0046] 1. Isolation and purification of Pseudomonas X2
[0047] Fresh rhizosphere soil from corn was taken, and under aseptic conditions, it was shaken and diluted with sterile water. The mixture was then spread on 50% TSB solid agar plates. After repeated isolation and purification, single colonies were obtained, which were identified as Pseudomonas X2.
[0048] 2. Identification of Pseudomonas X2
[0049] When Pseudomonas X2 was cultured on TSB solid medium for 48 hours, the colonies were yellow, round, smooth and moist, and Gram-negative.
[0050] Using Pseudomonas X2 DNA as a template, PCR amplification was performed using universal 16S rDNA primers 27F and 1492R. Primer 27F was 5'-AGAGTTTGATCCTGGCTCAG-3'; primer 1492R was 5'-GGTTACCTTGTTACGACTT-3'. Sequencing was performed after amplification, and the 16S rDNA sequence is shown in SEQ ID NO: 1 of the sequence listing. The obtained 16S rDNA sequencing results of Pseudomonas X2 were entered into the NCBI database for BLAST alignment to obtain the relevant type strain. After preliminary processing of the target sequence and the type strain sequence using MEGA11, a phylogenetic tree was constructed according to the Neighbor-joining method. The results are shown below. Figure 2 As shown, the 16S rRNA gene of Pseudomonas X2 is highly similar to that of Pseudomonas sp., therefore Pseudomonas X2 is identified as Pseudomonas sp., belonging to the genus Pseudomonas.
[0051] Example 3:
[0052] The preparation method of the compound microbial agent in this embodiment includes the following steps:
[0053] I. Activation of bacteria: Enterobacter X1 and Pseudomonas X2, which were stored in cold storage, were streaked onto TSB solid medium to obtain two activated strains.
[0054] II. Preparation of bacterial suspensions: Two activated bacterial strains were picked up with an inoculation needle and inoculated into TSB liquid medium to obtain Enterobacter X1 bacterial suspension and Pseudomonas X2 bacterial suspension.
[0055] III. Pretreatment of compound carriers: Wood ash was adjusted to pH 7.0 with 1M HCl, rinsed with deionized water until conductivity <1ms / cm, dried at 60℃ and then sterilized by high-pressure steam at 121℃ for 30min; Vermiculite was activated by soaking in 0.1M CaCl2 solution for 2h, dried at 60℃ until moisture content <5%, sterilized by dry heat at 160℃ for 2h, and cooled for later use.
[0056] IV. Preparation of the composite carrier: The wood ash treated in step three and vermiculite are mixed at a volume ratio of 1:1 to obtain a mixed matrix; sodium carboxymethyl cellulose (binder), trehalose (protectant), and humic acid (microbial activity promoter) are added to the mixed matrix to optimize the physicochemical properties and microbial compatibility of the carrier, thus obtaining the composite carrier; wherein the mass fractions of sodium carboxymethyl cellulose, trehalose, and humic acid in the composite carrier are 1%, 5%, and 0.5%, respectively.
[0057] V. Preparation of Compound Microbial Agent: Enterobacter X1 bacterial suspension and Pseudomonas X2 bacterial suspension were mixed at a volume ratio of 1:1 to form a compound bacterial solution. The compound bacterial solution was then mixed with a compound carrier at a volume ratio of 1:1 and air-dried at 30°C until the moisture content reached 20%. The carrier was then aged at room temperature for 48 hours, and the final moisture content was adjusted to 25%, thus obtaining the compound microbial agent.
[0058] A photograph of the composite microbial agent prepared in this embodiment is shown below. Figure 3 As shown. The compound microbial agent prepared in this embodiment was subjected to the following field trials.
[0059] The field experiment was conducted at the field experimental site of the Red Soil Ecological Experimental Station of the Chinese Academy of Sciences, located in Gongzhuling City, Jilin Province, China (124°8′N, 43°5′E). The soil pH in this area was 6.9, EC value was 77.29 μs / cm, soil organic carbon (SOC) content was 10.23 g / kg, total nitrogen (TN) content was 0.93 g / kg, and total phosphorus (TP) content was 0.50 g / kg. Each experimental plot was 20 × 5 m, belonging to a maize planting system. Each treatment was replicated three times. The compound microbial agent prepared in Example 2 was applied to the maize roots at a rate of 400 kg / ha. The control treatment did not receive any microbial agent.
[0060] (1) At maturity, the results of statistical analysis of the physicochemical properties of maize rhizosphere soil (including soil pH, EC, SOC, TN, and TP) are as follows: Figures 4-8 As shown in the figure. Statistical analysis indicates that the rhizosphere soil physicochemical environment was significantly optimized in the experimental group treated with the compound microbial agent of this invention. Compared with the control group, there were no significant differences in soil pH and EC, but SOC, TN, and TP all increased significantly, by 10.03%, 102.13%, and 56.26%, respectively. The compound microbial agent of this invention significantly optimized the rhizosphere microenvironment of mature maize by increasing SOC.
[0061] (2) At maturity, the growth status of corn was statistically analyzed, and the results are as follows: Figures 9-11 As shown in the figure, compared with the control, the fresh weight of maize plants increased by 43.0%, the dry weight increased by 42.0%, and the plant height increased by 15.8% after the application of compound microbial inoculant. This indicates that the compound microbial inoculant can promote maize growth.
[0062] (3) During the harvest season, the yield of corn was statistically analyzed. Compared with the control group, the yield increased significantly by 16.51% after adding the compound microbial agent, achieving the goal of increasing corn yield and income. The results are as follows: Figure 12 As shown.
Claims
1. A compound microbial agent for enhancing corn production capacity, characterized in that, The compound microbial agent includes a compound bacterial solution and a compound carrier. The compound bacterial solution includes Enterobacter suspension and Pseudomonas suspension. The compound carrier includes wood ash, vermiculite, sodium carboxymethyl cellulose, trehalose, and humic acid.
2. The compound microbial agent for improving corn production capacity according to claim 1, characterized in that, The Enterobacter sp. X1 was deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, on January 6, 2026, with accession number CCTCC NO: M 2026023.
3. A compound microbial agent for improving corn production capacity according to claim 1 or 2, characterized in that, The Pseudomonas species described is Pseudomonas sp. X2, deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, on January 6, 2026, with accession number CCTCC NO: M 2026024.
4. The method for preparing the compound microbial agent for improving corn production capacity as described in claim 1, characterized in that, The method includes the following steps:
1. Enterobacteriaceae and Pseudomonas were streaked onto TSB solid medium to obtain two activated strains.
2. Using an inoculation needle, two activated bacterial strains were picked up and inoculated separately into TSB liquid medium to obtain Enterobacter suspension and Pseudomonas suspension; the OD values of the two bacterial suspensions were measured. 600 The value is 0.8~1.0; 3. Dry the wood ash and sterilize it to obtain treated wood ash; soak the vermiculite in CaCl2 solution, dry it until the moisture content is <5%, sterilize it, and obtain treated vermiculite; 4. Mix the treated wood ash from step 3 with the treated vermiculite at a volume ratio of 1:1 to obtain a mixed matrix; add sodium carboxymethyl cellulose, trehalose, and humic acid to the mixed matrix to obtain a compound carrier; 5. Mix the Enterobacter suspension and Pseudomonas suspension to form a compound bacterial solution; mix the compound bacterial solution with the compound carrier at a volume ratio of 1:1, air dry, and then mature at room temperature. Finally, adjust the moisture content to 25% to obtain the compound microbial agent.
5. The method for preparing the compound microbial agent for improving corn production capacity according to claim 4, characterized in that, In step three, the pH of the wood ash is 7.0 and the conductivity is <1 ms / cm.
6. The method for preparing the compound microbial agent for improving corn production capacity according to claim 4 or 5, characterized in that, In step three, the concentration of the CaCl2 solution is 0.05~0.15 mol / L.
7. The method for preparing the compound microbial agent for improving corn production capacity according to claim 4, characterized in that, In step three, the mass fractions of sodium carboxymethyl cellulose, trehalose, and humic acid in the compound carrier are 1%, 5%, and 0.5%, respectively.
8. The application of the compound microbial agent as described in claim 1 in soil improvement.
9. The application of the compound microbial agent as described in claim 1 in promoting maize growth.
10. The application of the compound microbial agent as described in claim 1 in improving corn yield.