Microbial compound fertilizer for improving perilla yield and application thereof

Abstract

This invention discloses a microbial compound fertilizer for increasing perilla yield and its application, belonging to the field of microbial fertilizer technology. The microbial compound fertilizer comprises the following raw materials: compound microbial inoculant, humic acid, organic matter, trace elements, polyglutamic acid, and trehalose; the compound microbial inoculant includes *Bacillus polymyxa*, *Candida*, and *Penicillium oxalate*. This invention provides a microbial compound fertilizer that can significantly increase perilla yield. This microbial compound fertilizer integrates active ingredients such as organic matter, trace elements, and microorganisms. Through the scientific and rational compounding of these components, a stable synergistic effect is established among them, improving the soil environment and increasing fertilizer utilization, thereby effectively increasing perilla yield. It solves the problems of unstable microbial activity, insufficient fertilizer effect sustainability, and limited soil improvement effect in existing perilla cultivation, providing reliable technical support for high-quality and high-yield perilla cultivation.

Description

Technical Field

[0001] This invention relates to the field of microbial fertilizer technology, and in particular to a microbial compound fertilizer for increasing perilla yield and its application. Background Technology

[0002] Perilla (Perilla frutescens) is a plant used for both medicinal and edible purposes, possessing high medicinal and economic value. Currently, perilla cultivation commonly suffers from low yield, unstable quality, and low fertilizer utilization. Although various perilla-specific fertilizers and microbial fertilizers have been reported in existing technologies, and these fertilizers have played a positive role in improving perilla yield and quality to some extent, they still have significant shortcomings. For example, the microbial activity is unstable, making it difficult to maintain its effect, resulting in insufficient sustained fertilizer efficacy, and its soil-improving effect is also very limited.

[0003] Fulvic acid (FA), also known as fulvic acid, is an organic substance in humic acids, widely found in soil, water bodies, and sediments. Fulvic acid, with its relatively low molecular weight, is characterized by low carbon content and high oxygen content. It contains abundant acidic functional groups such as carboxyl, phenolic hydroxyl, and ketone carbonyl groups, making it strongly acidic in aqueous solutions. It can form complexes with metal ions (such as calcium, magnesium, and iron) and organic matter, affecting the migration and transformation of substances in the environment. In soil ecology, fulvic acid participates in aggregate formation as an organic colloid and activates trace elements through its carboxyl groups, promoting nutrient absorption by plants. However, there are currently no reports of compound fertilizers combining fulvic acid with specific microbial agents and designed specifically for the growth characteristics of perilla. Summary of the Invention

[0004] The purpose of this invention is to provide a microbial compound fertilizer for increasing perilla yield and its application, thereby solving the problems existing in the prior art. This invention provides a microbial compound fertilizer that can significantly increase perilla yield. This microbial compound fertilizer integrates active ingredients such as organic matter, trace elements, and microorganisms. Through the scientific and rational compounding of these components, a stable synergistic effect is established among them, improving the soil environment and increasing fertilizer utilization, thus effectively increasing perilla yield. It solves the problems of unstable microbial activity, insufficient fertilizer effect sustainability, and limited soil improvement effect in existing perilla cultivation, providing reliable technical support for high-quality and high-yield perilla cultivation.

[0005] To achieve the above objectives, the present invention provides the following solution: This invention provides a microbial compound fertilizer for increasing perilla yield. By weight, the microbial compound fertilizer comprises the following raw materials: 3-8 parts of compound microbial agent, 5-12 parts of fulvic acid, 40-60 parts of organic matter, 8-15 parts of trace elements, 0.1-0.5 parts of polyglutamic acid, and 0.1-0.3 parts of trehalose. The compound microbial agent contains Bacillus polymyxa, Candida albicans, and Penicillium oxalate.

[0006] Furthermore, the ratio of viable counts of the *Bacillus polymyxa*, *Candida*, and *Penicillium oxalate* is (60-80):(40-60):1; the total viable count of the composite microbial agent is 2-8 × 10⁻⁶. 10 CFU / mL.

[0007] Furthermore, the trace elements are composed of calcium magnesium phosphate fertilizer, zinc sulfate, boric acid and sodium molybdate mixed in a mass ratio of (5-8):(1-2):(1-2):(0.5-1).

[0008] Furthermore, the fulvic acid is mineral-derived fulvic acid.

[0009] Furthermore, the organic matter is a mixture of peat moss, biochar and straw in a mass ratio of (5-8):1:(10-20).

[0010] This invention also provides a method for preparing the above-mentioned microbial compound fertilizer, comprising the following steps: (1) Mix peat, biochar and straw and then crush them to obtain organic matter; (2) Mix Bacillus polymyxa, Candida albicans and Penicillium oxalate to obtain a compound microbial agent; (3) After mixing the organic matter with polyglutamic acid and trehalose, the compound microbial agent is inoculated, and after composting, trace elements and fulvic acid are added and mixed evenly. After granulation and drying, the microbial compound fertilizer is obtained.

[0011] Optionally, the particle size of the pulverized material is 0.5-2 cm; and the moisture content of the organic matter is 10%-15%.

[0012] Furthermore, the composting treatment is carried out at a temperature of 38-42℃ for 5-7 days; after 2-3 days of composting, the pile is turned over, and then turned over once a day thereafter.

[0013] The present invention also provides an application of the above-mentioned microbial compound fertilizer in increasing the yield of perilla.

[0014] The present invention also provides a method for increasing the yield of perilla: 7-10 days before sowing perilla, urea, superphosphate, potassium sulfate and the above-mentioned microbial compound fertilizer are applied as base fertilizer; wherein, the application rate of urea is 20-25 kg / mu, the application rate of superphosphate is 30-40 kg / mu, the application rate of potassium sulfate is 3-6 kg / mu, and the application rate of microbial compound fertilizer is 10-15 kg / mu.

[0015] The present invention discloses the following technical effects: This invention provides a microbial compound fertilizer that can significantly increase perilla yield. After using this compound fertilizer, seed yield can increase by 22.63%-27.42%. This microbial compound fertilizer integrates organic matter, trace elements, and microorganisms, among other active ingredients. Through the scientific and rational compounding of these components, a stable synergistic effect is established, improving the soil environment and increasing fertilizer utilization, thereby effectively increasing perilla yield. It solves the problems of unstable microbial activity, insufficient fertilizer effectiveness, and limited soil improvement in existing perilla cultivation methods, providing reliable technical support for high-quality and high-yield perilla cultivation. Detailed Implementation

[0016] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0017] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0018] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0019] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be readily apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0020] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0021] The research and development concept of this invention is as follows: To address the problems of unstable microbial activity, insufficient sustained fertilizer effect, and weak specificity for perilla in existing fertilizers, this invention aims to develop a perilla-specific microbial fertilizer with complementary components and synergistic effects. This microbial compound fertilizer integrates multiple active ingredients such as organic matter, trace elements, and microorganisms. Through the scientific and rational compounding of these components, a stable synergistic effect is established among them, effectively increasing perilla yield by improving the soil environment and enhancing fertilizer utilization.

[0022] This invention, through systematic strain screening and compounding experiments, ultimately determined that a composite microbial agent should be constructed with *Bacillus polymyxa*, *Candida*, and *Penicillium oxalate* as its core components. On one hand, these strains work synergistically to effectively regulate the soil microbial community structure, inhibit the growth and reproduction of harmful microorganisms, promote nutrient transformation in the soil, making it easier for perilla to absorb and utilize, and enhance the perilla's stress resistance, creating a favorable microbial environment for perilla growth, thereby promoting perilla growth and development and increasing yield. On the other hand, the strains in the composite microbial agent can decompose organic matter, releasing its nutrients. Their metabolites can also interact with fulvic acid, further enhancing the fulvic acid's effect of activating soil nutrients; while fulvic acid provides suitable environmental conditions for microbial growth and reproduction, helping the microbial agent to fully exert its effects. Organic matter not only provides basic nutrition and a suitable growth environment for microorganisms, effectively maintaining the activity of microbial fertilizers and prolonging their effects, but also enhances the environmental adaptability of microorganisms through pre-composting treatment. Combined with fulvic acid and trace elements, it improves the soil's physical and chemical properties and increases soil fertility. Micronutrients fully meet the nutrient requirements for perilla growth, working synergistically with microbial agents and humic acid to promote efficient absorption and utilization of nutrients by perilla. Polyglutamic acid and trehalose assist other components from the perspectives of water and fertilizer retention and enhancing stress resistance, respectively, ultimately achieving a significant improvement in overall fertilizer efficiency.

[0023] The strains used in the following examples can be purchased from the China Industrial Microbial Culture Collection Center (CICC) or the China Agricultural Microbial Culture Collection Center (ACCC). Fulvic acid can be purchased from Shandong Rongyao Biotechnology Co., Ltd.

[0024] Example 1 This embodiment provides a microbial compound fertilizer for increasing perilla yield. The raw material composition, by weight, is as follows: 5 parts compound microbial inoculant, 8 parts mineral-derived fulvic acid, 50 parts organic matter, 12 parts trace elements, 0.3 parts polyglutamic acid, and 0.2 parts trehalose. The compound microbial inoculant consists of *Bacillus polymyxa*, *Candida*, and *Penicillium oxalate*, with a viable count ratio of 70:50:1. The total viable count of the compound microbial inoculant is 5 × 10⁻⁶. 10CFU / mL; organic matter is a mixture of peat moss, biochar and straw in a mass ratio of 6:1:15; micronutrients are a mixture of calcium magnesium phosphate fertilizer, zinc sulfate, boric acid and sodium molybdate in a mass ratio of 6:1.5:1.5:0.5.

[0025] The preparation method of microbial compound fertilizer includes the following steps: (1) Organic matter preparation: Peat moss, biochar and straw are mixed in proportion, crushed to a particle size of 1 cm, and then the moisture content is adjusted to 12% to obtain organic matter for later use; (2) Preparation of compound microbial inoculant: Bacillus polymyxa, Candida albicans, and Penicillium oxalate were activated and cultured separately using NA medium, YPD medium, and PDA medium, respectively. After activation and culture, the bacterial cells were collected by centrifugation, resuspended in sterile water to obtain a bacterial suspension, and then mixed according to the ratio of viable bacteria to obtain the compound microbial inoculant. The total viable bacteria count of the compound microbial inoculant was 5 × 10⁻⁶. 10 CFU / mL; (3) Mixed composting: Mix the prepared organic matter with polyglutamic acid and trehalose evenly, then inoculate with compound microbial agent, control the fermentation temperature to 38-42℃ for composting treatment, turn the pile for the first time on the second day of composting, and then turn the pile once a day for 5 days to obtain composted material. (4) Finished product processing: Take out the decomposed material, add the medium and trace elements and mineral fulvic acid mixed in proportion, stir at low speed for 15 minutes until fully mixed, then send it to the granulator to granulate, control the particle size to 2-3 mm, and then place it in a 45℃ low temperature drying equipment to dry until the moisture content is ≤12%. After sieving through a 10-mesh sieve, package it to obtain the finished microbial compound fertilizer.

[0026] Example 2 This embodiment provides a microbial compound fertilizer for increasing perilla yield. The raw material composition, by weight, is as follows: 3 parts compound microbial inoculant, 5 parts mineral-derived fulvic acid, 60 parts organic matter, 8 parts trace elements, 0.1 parts polyglutamic acid, and 0.1 parts trehalose. The compound microbial inoculant consists of *Bacillus polymyxa*, *Candida*, and *Penicillium oxalate*, with a viable count ratio of 60:60:1. The total viable count of the compound microbial inoculant is 2 × 10⁻⁶. 10 CFU / mL; organic matter is a mixture of peat moss, biochar and straw in a mass ratio of 5:1:10; micronutrients are a mixture of calcium magnesium phosphate fertilizer, zinc sulfate, boric acid and sodium molybdate in a mass ratio of 5:2:2:0.5.

[0027] The preparation method of microbial compound fertilizer includes the following steps: (1) Organic matter preparation: Peat moss, biochar and straw are mixed in proportion, crushed to a particle size of 0.5 cm, and the moisture content is adjusted to 10% to obtain organic matter for later use; (2) Preparation of compound microbial inoculant: Bacillus polymyxa, Candida albicans, and Penicillium oxalate were activated and cultured separately using NA medium, YPD medium, and PDA medium, respectively. After activation and culture, the bacterial cells were collected by centrifugation, resuspended in sterile water to obtain a bacterial suspension, and then mixed according to the ratio of viable bacteria to obtain the compound microbial inoculant. The total viable bacteria count of the compound microbial inoculant was 2 × 10⁻⁶. 10 CFU / mL; (3) Mixed composting: Mix the prepared organic matter with polyglutamic acid and trehalose evenly, then inoculate with compound microbial agent, control the temperature at 38-42℃ for composting, turn the pile for the first time on the 3rd day of composting, and then turn the pile once a day for 7 days to obtain the composted material. (4) Finished product processing: Add trace elements and mineral fulvic acid to the decomposed material, stir for 10 minutes to mix evenly, then send it to a granulator to granulate, control the particle size to 2-3 mm, and then place it in a 45℃ low temperature drying equipment to dry to a moisture content ≤12%. After sieving through a 10-mesh sieve, package it to obtain the finished microbial compound fertilizer.

[0028] Example 3 This embodiment provides a microbial compound fertilizer for increasing perilla yield. The raw material composition, by weight, is as follows: 8 parts compound microbial inoculant, 12 parts mineral-derived fulvic acid, 40 parts organic matter, 15 parts trace elements, 0.5 parts polyglutamic acid, and 0.3 parts trehalose. The compound microbial inoculant consists of *Bacillus polymyxa*, *Candida*, and *Penicillium oxalate*, with a viable count ratio of 80:40:1. The total viable count of the compound microbial inoculant is 8 × 10⁻⁶. 10 CFU / mL; organic matter is a mixture of peat moss, biochar and straw in a mass ratio of 8:1:20; micronutrients are a mixture of calcium magnesium phosphate fertilizer, zinc sulfate, boric acid and sodium molybdate in a mass ratio of 8:1:1:1.

[0029] The preparation method of microbial compound fertilizer includes the following steps: (1) Organic matter preparation: Mix peat, biochar and straw in proportion, crush to a particle size of 2cm, adjust the moisture content to 15%, and obtain organic matter for later use; (2) Preparation of compound microbial inoculant: Bacillus polymyxa, Candida albicans, and Penicillium oxalate were activated and cultured separately using NA medium, YPD medium, and PDA medium, respectively. After activation and culture, the bacterial cells were collected by centrifugation, resuspended in sterile water to obtain a bacterial suspension, and then mixed according to the ratio of viable bacteria to obtain the compound microbial inoculant; the total viable bacteria count of the compound microbial inoculant was 8 × 10⁻⁶. 10 CFU / mL; (3) Mixed composting: Mix the prepared organic matter with polyglutamic acid and trehalose evenly, then inoculate with compound microbial agent, control the temperature at 38-42℃ for composting, turn the pile for the first time on the second day of composting, and then turn the pile once a day for 5 days to obtain composted material. (4) Finished product processing: Add trace elements and mineral fulvic acid to the decomposed material, stir for 20 minutes to mix evenly, then send it to a granulator to granulate, control the particle size to 2-3 mm, and then place it in a 45℃ low temperature drying equipment to dry to a moisture content ≤12%. After sieving through a 10-mesh sieve, package it to obtain the finished microbial compound fertilizer.

[0030] Comparative Example 1 Same as Example 1, except that the compound microbial agent is replaced with a polymyxin Bacillus agent, while the total number of viable bacteria remains unchanged.

[0031] Comparative Example 2 Same as Example 1, except that the compound microbial agent is replaced with Penicillium oxalate agent, while the total number of viable bacteria remains unchanged.

[0032] Comparative Example 3 Same as Example 1, except that Penicillium oxalate in the compound microbial agent is replaced with Trichoderma harzianum, while the ratio of viable counts of each species and the total viable count remain unchanged.

[0033] Comparative Example 4 Same as Example 1, except that the compound microbial agent is omitted.

[0034] Comparative Example 5 Same as Example 1, except that fulvic acid is omitted.

[0035] Experimental Example 1 The effects of the microbial compound fertilizers prepared in Examples 1-3 and Comparative Examples 1-5 on the growth of perilla were verified: A small-plot experiment was conducted at the Kangjin Experimental Base in Hulan District, Harbin City, Heilongjiang Province. A total of nine plots were set up. Except for the blank control group, the other eight plots used the microbial compound fertilizers prepared in Examples 1-3 and Comparative Examples 1-5, respectively. Each plot had a depth of 15m³. 2 Using "Longzisu No. 1" as the experimental variety, 40 plants were planted in each plot.

[0036] Seven days before sowing perilla, each plot received 22 kg of urea, 35 kg of superphosphate, and 5 kg of potassium sulfate per mu (approximately 0.067 hectares) during land preparation and fertilization, along with 12 kg of the microbial compound fertilizer described in this example. The fertilizer was thoroughly mixed with the soil at a depth of 30 cm, and the plots were left to stand for 7 days before sowing. The control group received only urea, superphosphate, and potassium sulfate. After sowing, the perilla was cultivated according to local conventional management methods until maturity. The perilla yield of each plot was statistically analyzed and is shown in Table 1.

[0037] Table 1. Statistical results of perilla yield in each plot As shown in Table 1, the microbial compound fertilizers of Examples 1-3 significantly increased perilla yield, with seed yield increasing by 22.63%-27.42% compared to the blank control group. In contrast, Comparative Examples 1-2, using a single microbial agent, only achieved a yield increase of 0.62%-8.70%, indicating that microbial strains adapted to the growth characteristics of perilla must be selected to achieve the desired yield increase. Comparative Example 3, after replacing the microbial strains in the compound microbial agent, showed a lower yield increase than the examples, confirming the synergistic effect of *Bacillus polymyxa*, *Candida*, and *Penicillium oxalate*. Comparative Example 4, omitting the compound microbial agent, only saw a yield increase of 3.37%, and Comparative Example 5, omitting mineral-derived fulvic acid, saw a yield increase of 9.23%, both significantly lower than the examples, indicating that fulvic acid has a synergistic effect with the microbial agent, organic matter, and other components. In summary, the complementary and synergistic effects of the components in the microbial compound fertilizer of this invention are key to high perilla yield; the absence or replacement of a single component will disrupt the balance and significantly reduce fertilizer efficiency.

[0038] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A microbial compound fertilizer for increasing perilla yield, characterized in that, By weight, the microbial compound fertilizer contains the following raw materials: 3-8 parts of compound microbial agent, 5-12 parts of humic acid, 40-60 parts of organic matter, 8-15 parts of trace elements, 0.1-0.5 parts of polyglutamic acid, and 0.1-0.3 parts of trehalose; The compound microbial agent contains Bacillus polymyxa, Candida albicans, and Penicillium oxalate.

2. The microbial compound fertilizer according to claim 1, characterized in that, The ratio of viable counts of the *Bacillus polymyxa*, *Candida*, and *Penicillium oxalate* is (60-80):(40-60):1; the total viable count of the compound microbial agent is 2-8 × 10⁻⁸. 10 CFU / mL.

3. The microbial compound fertilizer according to claim 1, characterized in that, The micronutrients are composed of calcium magnesium phosphate fertilizer, zinc sulfate, boric acid and sodium molybdate in a mass ratio of (5-8):(1-2):(1-2):(0.5-1).

4. The microbial compound fertilizer according to claim 1, characterized in that, The fulvic acid mentioned is mineral-derived fulvic acid.

5. The microbial compound fertilizer according to claim 1, characterized in that, The organic matter is a mixture of peat, biochar and straw in a mass ratio of (5-8):1:(10-20).

6. A method for preparing a microbial compound fertilizer as described in any one of claims 1-5, characterized in that, Includes the following steps: (1) Mix peat, biochar and straw and then crush them to obtain organic matter; (2) Mix Bacillus polymyxa, Candida albicans and Penicillium oxalate to obtain a compound microbial agent; (3) After mixing the organic matter with polyglutamic acid and trehalose, the compound microbial agent is inoculated, and after composting, trace elements and fulvic acid are added and mixed evenly. After granulation and drying, the microbial compound fertilizer is obtained.

7. The preparation method according to claim 6, characterized in that, The particle size of the pulverized material is 0.5-2 cm; the moisture content of the organic matter is 10%-15%.

8. The preparation method according to claim 6, characterized in that, The composting treatment is carried out at a temperature of 38-42℃ for 5-7 days. After 2-3 days of composting, the compost pile is turned over, and then turned over once a day thereafter.

9. The application of a microbial compound fertilizer as described in any one of claims 1-5 in increasing perilla yield.

10. A method for increasing perilla yield, characterized in that, 7-10 days before sowing perilla, apply urea, superphosphate, potassium sulfate, and the microbial compound fertilizer according to any one of claims 1-5 as base fertilizer; wherein the application rate of urea is 20-25 kg / mu, the application rate of superphosphate is 30-40 kg / mu, the application rate of potassium sulfate is 3-6 kg / mu, and the application rate of microbial compound fertilizer is 10-15 kg / mu.

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