Microbial source humic acid fertilizer for promoting glycerol secretion of oil bean root

Abstract

The present application relates to the promotion oil bean root secretion of mevalonic acid microbial source humic acid fertilizer, the promotion oil bean root secretion of mevalonic acid artificial synthesis microbial source humic acid is prepared by the following method: the corn stalk is predecomposed by using the big ball cover mushroom strain, and the corn slurry with the concentration of 0-1% is added, the moisture content of corn stalk is 55-65%, the predecomposed corn stalk is added with water, is handled with water treatment device, and the mixture containing microbial fungal residues is obtained;The mixture containing microbial fungal residues is converted into artificial synthesis microbial source humic acid by four-stage reaction;The artificial synthesis microbial source humic acid fertilizer is applied to the root of oil bean, the mevalonic acid secreted by the root of oil bean reaches 0.006-0.007 ng / mg, and the length of the root of oil bean reaches 25-26 cm.The present application can improve the content of mevalonic acid secreted by the root of oil bean, improve the yield and quality of oil bean.

Description

Technical Field

[0001] This invention relates to microbial fertilizer manufacturing technology, specifically to a method for preparing and applying artificially synthesized microbial humic acid that can enhance the secretion of mevalonic acid by the roots of cowpeas. Background Technology

[0002] Green beans thrive in warm, dry climates with ample sunlight. They are susceptible to frost and heat, and are best suited for loam or sandy loam soils. The optimal temperature for growth and development is 18-26℃. Seeds will not germinate below 8℃ or above 38℃, seedlings will cease growth below 13℃, and flowering and pod-setting will be less efficient below 15℃ or above 30℃. Sowing is typically done in spring when local temperatures are consistently above 15℃. Direct sowing or transplanting seedlings can be used. For direct sowing, scatter the seeds evenly on the soil, cover with a thin layer of soil, and gently compact. For transplanting seedlings, first raise the seeds in seedling trays, and then transplant them into the field once they have reached a certain height. Field management and watering: Green beans require relatively high moisture levels. Keep the soil moist, increase watering during dry seasons, and ensure proper drainage during rainy seasons to prevent waterlogging and root rot. Fertilization: Apply top dressing as needed during the growing season, using compound fertilizer or organic fertilizer every 10-15 days to promote plant growth and fruit development. Weeding and Soil Loosening: Weed promptly to keep the soil loose, and loosen the soil regularly to increase soil aeration, which is beneficial for root growth. Pest and Disease Control: Adopt integrated management measures, including physical, biological, and chemical methods, to control the occurrence and spread of pests and diseases in a timely manner.

[0003] Phosphorus fixation in soil has long been a key limiting factor for crop growth. Soil phosphorus exists in various forms, including inorganic and organic phosphorus. Inorganic phosphorus accounts for 50%-80% of total soil phosphorus, while organic phosphorus accounts for 10%-15%. Inorganic phosphorus dominates because, after fixation, most of it exists in the soil in adsorbed and solid mineral forms, making it difficult for plant roots to effectively absorb. Its availability is significantly lower than that of organic phosphorus, which easily inhibits the normal growth and development of crops, ultimately having an adverse effect on plant growth. The secretion of organic acids by roots can increase the availability of phosphorus in the soil, thereby promoting crop growth.

[0004] Traditional cultivation of green beans suffers from low levels of organic acids secreted by the roots, which affects the availability of soil nutrients during cultivation, thus impacting the yield and quality of the green beans. Summary of the Invention

[0005] The purpose of this invention is to provide a microbial humic acid fertilizer that promotes the secretion of mevalonic acid by the roots of cowpeas. This artificially synthesized microbial humic acid fertilizer that promotes the secretion of mevalonic acid by the roots of cowpeas is used to solve the problem that the root secretion of organic acids in traditional cowpea cultivation is low, which affects the availability of soil nutrients during cowpea cultivation and thus affects the yield and quality of cowpeas.

[0006] To achieve the above objectives, the specific technical solution adopted by the present invention is as follows: This artificially synthesized microbial humic acid that promotes the secretion of mevalonic acid by the roots of cowpeas is prepared by the following method:

[0007] First, corn stalks are pre-decomposed using Pleurotus ostreatus spawn, and corn syrup with a concentration of 0.5-1% is added. The moisture content of the corn stalks is 55-65%. After pre-decomposition, water is added to the corn stalks, and the mixture is processed by a water processor to obtain a mixture containing microbial and fungal residues.

[0008] A mixture containing microbial and fungal residues undergoes a four-stage reaction to be converted into artificially synthesized microbial humic acid:

[0009] Potassium hydroxide and sodium hydroxide were added as catalysts to a mixture containing microbial and fungal residues at a mass ratio of 8:1. The reaction temperature was controlled at 140℃ for the first stage reaction, which lasted for 1 hour. Potassium hydroxide and sodium hydroxide were added again as catalysts at a mass ratio of 8:1, and the reaction temperature was controlled at 160℃ for the second stage reaction, which lasted for 1 hour. Next, potassium permanganate, potassium sulfate, and copper sulfate were added as catalysts at a mass ratio of 10:10:1, and the reaction temperature was controlled at 180℃ for the third stage reaction, which lasted for 1 hour. Finally, potassium hydroxide was added again as a catalyst, and the reaction temperature was controlled at 200℃ for the fourth stage reaction, which lasted for 2 hours. The resulting synthetic microbial humic acid liquid was dried to obtain synthetic microbial humic acid fertilizer.

[0010] The artificially synthesized microbial humic acid fertilizer was diluted with water and applied to the roots of the cowpea seedlings. After the seedling stage, the mevalonic acid secreted by the cowpea roots reached 0.006-0.007 ng / mg, and the length of the cowpea roots reached 25-26 cm.

[0011] In the above scheme, the pre-decomposition temperature of corn stalks is 15-32℃, the pre-decomposition time is 25-45 days, and the mass ratio of the pre-decomposed corn stalks to the volume ratio of water is 1g:10-30ml.

[0012] In the above scheme, the water added to the pre-decomposed corn stalks is treated by a water processor at a frequency of 20000Hz for 10-30 minutes.

[0013] In the first and second stage reactions of the above scheme, the total mass ratio of potassium hydroxide and sodium hydroxide to the volume ratio of the mixture containing microbial and fungal residues is 0.5-1.5%.

[0014] In the above scheme, the total mass of potassium permanganate, potassium sulfate, and copper sulfate is 0.1-2% of the volume of the mixture containing microbial and fungal residues.

[0015] In the above scheme, the total mass of potassium hydroxide and the volume ratio of the mixture containing microbial and fungal residues in the fourth stage reaction are 0.5-1.5%.

[0016] In the above scheme, when artificially synthesized microbial humic acid is applied to the roots of cowpeas, it should be diluted with water 1000 to 3000 times, and the application amount after dilution should be 10 to 100 mL per plant. Beneficial effects

[0017] This invention increases the content of mevalonic acid secreted by the roots of cowpeas by applying artificially synthesized microbial humic acid, synthesized from corn stalks containing *Agaricus bisporus* mycelium. It also provides a method for the pre-decomposition of corn stalks and the artificial synthesis of microbial humic acid. Mevalonic acid, as an organic acid secreted by plant roots, has the function of improving soil nutrient availability.

[0018] 2. The active functional groups such as carboxyl groups (-COOH) and phenolic hydroxyl groups (-OH) in the artificially synthesized microbial humic acid prepared in this invention can stimulate cell metabolism and enhance the secretion level of mevalonic acid by interacting with root cells, thereby promoting the availability of soil nutrients. It can promote the growth of cowpea roots, making the length of cowpea roots reach 25-26 cm, while the mevalonic acid secreted by cowpea roots reaches 0.006-0.007 ng / mg.

[0019] 3. This invention utilizes *Stropharia macrocarpa* to degrade straw. *Stropharia macrocarpa* decomposes complex organic substances such as cellulose, hemicellulose, and lignin in corn straw through its mycelium. After decomposition, these organic substances are transformed into nutrients such as amino acids and vitamins. Attached Figure Description

[0020] Figure 1 A comparison chart showing the content of mevalonic acid secreted by the roots of green beans after applying this invention and water-grown beans.

[0021] Figure 2 A comparison diagram showing the length of the root system of a green bean (cowpea) after applying the present invention. Detailed Implementation

[0022] The invention will be further described below with reference to the accompanying drawings: Example

[0023] This artificially synthesized microbial humic acid, which enhances the secretion of mevalonic acid by the roots of cowpeas, is prepared by the following method: Corn stalks are pre-decomposed using *Agaricus bisporus* spawn. During pre-decomposition, the moisture content of the corn stalks is 55%, and 0.5% corn steep liquor is added. The pre-decomposition temperature is 32°C, and the pre-decomposition time is 25 days. The pre-decomposed corn stalks are then added to water treated by a water processor at 20000Hz for 10 minutes to obtain a mixture containing microbial fungal residues. This mixture undergoes a four-stage reaction to convert into artificially synthesized microbial humic acid. In the first stage, potassium hydroxide and sodium hydroxide are added as catalysts at a mass ratio of 8:1, and the total mass of potassium hydroxide and sodium hydroxide is 0.5% of the volume of the mixture. The reaction temperature is controlled at 140°C, and the reaction time is 1 hour. In the second stage, potassium hydroxide and sodium hydroxide are added as catalysts, with a mass ratio of 8:1 and a total mass ratio of 1.5% of the mixture. The reaction temperature is controlled at 160℃, and the reaction time is 1 hour. In the third stage, potassium permanganate, potassium sulfate, and copper sulfate are added as catalysts, with a mass ratio of 10:10:1 and a total mass ratio of 0.1% of the mixture. The reaction temperature is controlled at 180℃, and the reaction time is 1 hour. In the fourth stage, potassium hydroxide is added as a catalyst, with a total mass ratio of 0.5% of the mixture. The reaction temperature is controlled at 200℃, and the reaction time is 2 hours. The resulting liquid is dried to obtain synthetic microbial humic acid. When applying the synthetic microbial humic acid to the roots of cowpeas, it needs to be diluted 1000 times, with an application rate of 10 mL per plant.

[0024] Straw, as a biomass resource rich in cellulose, hemicellulose, and lignin, can be gradually decomposed into more readily usable small-molecule compounds through the metabolism of *Stropharia masticata* spores. During metabolism, *Stropharia masticata* spores not only break down straw into basic organic substances such as reducing sugars and proteins, but also accumulate abundant amino acids and other nitrogen-containing compounds during their own reproduction and growth. These compounds can effectively synthesize high-molecular-weight organic matter similar to humic acid, providing a new potential pathway for the efficient utilization of biomass resources and soil improvement. Example

[0025] This method and application for preparing and applying artificially synthesized microbial humic acid that can enhance the secretion of mevalonic acid by the roots of cowpeas involves: Pre-decomposing corn stalks using *Agaricus bisporus* spawn. The moisture content of the corn stalks during pre-decomposition is 65%, and 0.7% corn steep liquor is added. The pre-decomposition temperature is 25°C, and the pre-decomposition time is 35 days. The pre-decomposed corn stalks are then added to water treated by a water processor at 20000Hz for 20 minutes to obtain a mixture containing microbial fungal residues. This mixture undergoes a four-stage reaction to convert into artificially synthesized microbial humic acid. In the first stage, potassium hydroxide and sodium hydroxide are added as catalysts at a mass ratio of 8:1, and the total mass of potassium hydroxide and sodium hydroxide is 1.5% of the volume of the mixture. The reaction temperature is controlled at 140°C, and the reaction time is 1 hour. In the second stage, potassium hydroxide and sodium hydroxide are added as catalysts, with a mass ratio of 8:1 and a total mass ratio of 0.5% of the mixture. The reaction temperature is controlled at 160℃, and the reaction time is 1 hour. In the third stage, potassium permanganate, potassium sulfate, and copper sulfate are added as catalysts, with a mass ratio of 10:10:1 and a total mass ratio of 0.1% of the mixture. The reaction temperature is controlled at 180℃, and the reaction time is 1 hour. In the fourth stage, potassium hydroxide is added as a catalyst, with a total mass ratio of 0.5% of the mixture. The reaction temperature is controlled at 200℃, and the reaction time is 2 hours. The resulting liquid is dried to obtain synthetic microbial humic acid. When applying the synthetic microbial humic acid to the roots of cowpeas, it needs to be diluted 1000 times, with an application rate of 10 mL per plant. Example

[0026] This method and application for preparing and applying artificially synthesized microbial humic acid that can enhance the secretion of mevalonic acid by the roots of cowpeas involves: Pre-decomposing corn stalks using *Agaricus bisporus* spawn. The moisture content of the corn stalks during pre-decomposition is 60%, and 1% corn steep liquor is added. The pre-decomposition temperature is 20℃, and the pre-decomposition time is 35 days. The pre-decomposed corn stalks are then added to water treated by a water processor at 20000Hz for 10 minutes to obtain a mixture containing microbial fungal residues. This mixture can be converted into artificially synthesized microbial humic acid through a four-stage reaction. In the first stage, potassium hydroxide and sodium hydroxide are added as catalysts at a mass ratio of 8:1, and the total mass of potassium hydroxide and sodium hydroxide is 1.5% of the volume of the mixture. The reaction temperature is controlled at 140℃, and the reaction time is 1 hour. In the second stage, potassium hydroxide and sodium hydroxide are added as catalysts, with a mass ratio of 8:1 and a total mass of 0.5% of the mixture's volume. The reaction temperature is controlled at 160℃, and the reaction time is 1 hour. In the third stage, potassium permanganate, potassium sulfate, and copper sulfate are added as catalysts, with a mass ratio of 10:10:1 and a total mass of 2% of the mixture's volume. The reaction temperature is controlled at 180℃, and the reaction time is 1 hour. In the fourth stage, potassium hydroxide is added as a catalyst, with a mass of 0.5% of the mixture's volume. The reaction temperature is controlled at 200℃, and the reaction time is 2 hours. The resulting liquid is dried to obtain synthetic microbial humic acid. When applying the synthetic microbial humic acid to the roots of cowpeas, it needs to be diluted 3000 times, with an application rate of 100 mL per plant.

[0027] To verify the effectiveness of the present invention, a confidentiality experiment was conducted, as follows:

[0028] This experiment prepared artificially synthesized microbial humic acid fertilizer according to the method of the invention patent. First, corn stalks were pre-decomposed using *Agaricus bisporus* spawn, with the moisture content of the corn stalks controlled at 65%, and 0.5% corn steep liquor was added as a promoter. The pre-decomposition temperature was controlled at 20℃ for 30 days. The pre-decomposed corn stalks were then added to water that had been treated at a frequency of 20000Hz for 10-30 minutes, and mixed at a mass-to-volume ratio of 1g:20ml to obtain a pre-decomposed mixture containing microbial fungal residues. Next, the mixture containing microbial fungal residues undergoes a four-stage reaction to convert it into artificially synthesized microbial humic acid: In the first and second stages, potassium hydroxide and sodium hydroxide are added respectively (the mass ratio of potassium hydroxide to sodium hydroxide is 8:1, and the total mass of potassium hydroxide and sodium hydroxide to the volume ratio of the mixture is 1.5%), with the reaction temperatures controlled at 140℃ and 160℃, respectively, for 1 hour each; in the third stage, potassium permanganate, potassium sulfate, and copper sulfate are added (the mass ratio of potassium permanganate, potassium sulfate, and copper sulfate is 10:10:1, and the total mass of potassium permanganate, potassium sulfate, and copper sulfate to the volume ratio of the mixture is 2%), with the reaction temperature at 180℃ for 1 hour; in the fourth stage, potassium hydroxide is added (the mass ratio of potassium hydroxide to the volume ratio of the mixture is 0.5%), with the reaction temperature at 200℃ for 2 hours. After the reaction is completed, the liquid is dried to obtain artificially synthesized microbial humic acid fertilizer.

[0029] The experiment was validated using a soil-cultivated potted method for green bean seedlings. Green beans were planted in pots of nutrient-rich soil, with an experimental group and a control group. The experimental group received a synthetic microbial humic acid fertilizer diluted 1000 times, at a dosage of 50 mL per plant; the control group received only water. Greenhouse conditions were maintained throughout the experiment, with regular watering to ensure normal growth of the green beans. At the end of the seedling stage, the soil around the roots was removed, the roots of the green bean plants were thoroughly cleaned, and they were placed in purified water to collect root exudates. The collected exudate samples were transported under refrigeration to Meiji Biotechnology Co., Ltd., where the organic acid composition, particularly mevalonic acid, in the root exudates was analyzed using an organic acid detection method.

[0030] The experimental results show that, see Figure 1 Compared with applying water, treatment with synthetic microbial humic acid can increase the content of mevalonic acid secreted by the roots of cowpea, with the mevalonic acid secreted by the roots reaching 0.006-0.007 ng / mg.

[0031] The experimental results show that, see Figure 2 Compared with applying water, applying synthetic microbial humic acid can increase the root length of cowpeas, reaching 25-26 cm.

Claims

1. A microbial-derived humic acid fertilizer that promotes the secretion of mevalonic acid by the roots of cowpeas, characterized in that: The microbial-derived humic acid that promotes the secretion of mevalonic acid from the roots of cowpeas is prepared by the following method: First, corn stalks are pre-decomposed using Pleurotus ostreatus spawn, and corn syrup with a concentration of 0.5-1% is added. The moisture content of the corn stalks is 55-65%. After pre-decomposition, water is added to the corn stalks, and the mixture is processed by a water processor to obtain a mixture containing microbial and fungal residues. A mixture containing microbial and fungal residues undergoes a four-stage reaction to be converted into artificially synthesized microbial humic acid: Potassium hydroxide and sodium hydroxide were added as catalysts to a mixture containing microbial and fungal residues at a mass ratio of 8:

1. The reaction temperature was controlled at 140℃ for the first stage reaction, which lasted for 1 hour. Potassium hydroxide and sodium hydroxide were added again as catalysts at a mass ratio of 8:1, and the reaction temperature was controlled at 160℃ for the second stage reaction, which lasted for 1 hour. Next, potassium permanganate, potassium sulfate, and copper sulfate were added as catalysts at a mass ratio of 10:10:1, and the reaction temperature was controlled at 180℃ for the third stage reaction, which lasted for 1 hour. Finally, potassium hydroxide was added again as a catalyst, and the reaction temperature was controlled at 200℃ for the fourth stage reaction, which lasted for 2 hours. The resulting synthetic microbial humic acid liquid was dried to obtain synthetic microbial humic acid fertilizer. In the fourth stage reaction, the total mass ratio of potassium hydroxide to the volume ratio of the mixture containing microbial and fungal residues is 0.5-1.5%. The artificially synthesized microbial humic acid fertilizer was diluted with water and applied to the roots of the cowpea seedlings. After the seedling stage, the mevalonic acid secreted by the cowpea roots reached 0.006-0.007 ng / mg, and the length of the cowpea roots reached 25-26 cm.

2. The microbial humic acid fertilizer for promoting mevalonic acid secretion from the roots of cowpeas according to claim 1, characterized in that: The pre-decomposition temperature of the corn stalks is 15-32℃, the pre-decomposition time is 25-45 days, and the mass ratio of the pre-decomposed corn stalks to the volume of water is 1g:10-30ml.

3. The microbial humic acid fertilizer for promoting mevalonic acid secretion from the roots of cowpeas according to claim 2, characterized in that: The pre-decomposed corn stalks were added to water and treated with a water processor at a frequency of 20000Hz for 10-30 minutes.

4. The microbial humic acid fertilizer for promoting mevalonic acid secretion from the roots of cowpeas according to claim 3, characterized in that: During the first and second stage reactions, the total mass ratio of potassium hydroxide and sodium hydroxide to the volume ratio of the mixture containing microbial and fungal residues is 0.5-1.5%.

5. The microbial humic acid fertilizer for promoting mevalonic acid secretion from the roots of cowpeas according to claim 4, characterized in that: The total mass ratio of potassium permanganate, potassium sulfate, and copper sulfate to the volume ratio of the mixture containing microbial and fungal residues is 0.1-2%.

6. The microbial humic acid fertilizer for promoting mevalonic acid secretion from the roots of cowpeas according to claim 5, characterized in that: When the artificially synthesized microbial humic acid fertilizer is applied to the roots of cowpeas, it should be diluted with water 1000 to 3000 times, and the application amount after dilution should be 10 to 100 mL per plant.

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