Soil phosphorus-dissolving solid bacterial fertilizer
The soil phosphorus-soluble solid microbial fertilizer prepared by the combined strains of *Penicillium desertii* and *Penicillium fimbriatum* solves the problem of plants' difficulty in absorbing soil phosphorus, achieves a highly efficient phosphorus-soluble effect, promotes crop growth and improves soil phosphorus utilization.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-31
- Publication Date
- 2026-03-31
AI Technical Summary
Plants have difficulty absorbing phosphorus directly from the soil, resulting in low phosphorus utilization and limiting crop growth and development.
A composite strain of *Penicillium desertorum* and *Penicillium steckii* was used as a soil phosphorus-solubilizing bacterial fertilizer, utilizing agricultural waste substrate as a carrier to enhance phosphorus-solubilizing capacity through synergistic effects.
It significantly improves the solubility of soil phosphorus, promotes crop growth, enhances soil phosphorus utilization, and meets the standards for agricultural microbial agents.
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Figure CN121758221A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of microbial fertilizer technology, specifically relating to a soil phosphorus-soluble solid microbial fertilizer. Background Technology
[0002] Phosphorus, an essential mineral element for plant growth and development, participates in various metabolic processes such as photosynthesis, respiration, signal transduction, and energy metabolism. It is also a component of organic macromolecules like proteins and phospholipids within plants. However, due to phosphorus immobilization in the soil, plants cannot directly absorb and utilize it. Phosphorus deficiency thus becomes a significant limiting factor in plant growth and development. Phosphorus-solubilizing bacteria, as biological inoculants, can improve phosphorus availability, and their ability to convert insoluble phosphorus into soluble forms has been well-studied. Research indicates that fungi have a much higher phosphorus-solubilizing capacity than bacteria. Microbial inoculants, as novel environmentally friendly fertilizers, are rich in beneficial live bacteria and various natural active substances. In agricultural production, microbial inoculants can effectively improve soil structure, enhance crop resistance, and promote crop growth and development, thereby increasing crop yield and improving quality.
[0003] Currently, improving soil phosphorus use efficiency and promoting crop growth mainly relies on directly adding phosphate-solubilizing bacteria to the soil or applying inoculants. In-depth research into this technology is of great significance for improving soil phosphorus use efficiency and promoting healthy crop growth. Compared to traditional phosphate fertilizer application methods, phosphate-solubilizing inoculants have significant advantages such as no secondary pollution, strong sustainability, and high efficiency, demonstrating unique value in the agricultural field. Therefore, studying the application effects of phosphate-solubilizing inoculants is of great significance for improving agricultural soil quality, ensuring crop production safety, and achieving sustainable development of agricultural resources. Summary of the Invention
[0004] In view of this, the purpose of this invention is to provide a soil phosphorus-solidifying bacterial fertilizer. The bacterial fertilizer of this invention has a good phosphorus-solubilizing effect, and the phosphorus-solubilizing capacity is further improved through the synergistic effect of bacterial strains.
[0005] To achieve the above objectives, the present invention provides the following technical solution: This invention provides a soil phosphorus-soluble solid microbial fertilizer, comprising *Penicillium desertii* (… Penicillium desertorum ) and Penicillium fimbriatum ( Penicillium steckii ); The desert penicillium ( Penicillium desertorum It was deposited at the China General Microbiological Culture Collection Center on January 9, 2025, with accession number CGMCC NO: 41748; The Penicillium fissure ( Penicillium steckiiIt was deposited on January 8, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO: 41749.
[0006] Preferably, the ratio of *Penicillium desertii* to *Penicillium chrysogenum* is 1:1.
[0007] Preferably, the phosphorus-soluble solid microbial fertilizer for soil can reach a phosphorus soluble content of 984.43 mg / L.
[0008] Preferably, the soil phosphorus-soluble solid microbial fertilizer also includes agricultural waste microbial bran.
[0009] Preferably, the agricultural waste mushroom substrate is mixed with distilled water, and the ratio of agricultural waste mushroom substrate to distilled water is 1:3.
[0010] Preferably, the inoculum volume is 9 wt%.
[0011] Preferably, the soil phosphorus-soluble solid bacterial fertilizer has a pH of 8.18, a fineness of 80.0, a moisture content of 19.32%, and a viable bacterial count of 2.2 × 10⁻⁶. 8 CFU / g.
[0012] It contains at least the following beneficial technical effects: The microbial fertilizer described in this invention exhibits excellent phosphorus-solubilizing effects, with the synergistic effect of the bacterial strains further enhancing its phosphorus-solubilizing capacity. A phosphorus-solubilizing fungal agent was prepared using agricultural waste substrate as a carrier, with *Penicillium desertii* achieving a maximum sporulation yield of 5.56 × 10⁻⁶. 10 The highest sporulation rate of the compound bacterial strain reached 6.17 × 10⁶ cells / mL. 10 The solid fungal agent prepared under the conditions of a material-to-liquid ratio of 1:3 and an inoculum amount of 9% had a pH of 8.18, a fineness of 80.0, a water content of 19.32%, and a viable count of 2.2 × 10⁶ cells / mL. 8 The CFU / g values all meet the standards for agricultural microbial inoculants (GB20287-2006); the maximum phosphorus solubility after mixing the two fungi in a 1:1 ratio can reach 984.43 mg / L. Attached Figure Description
[0013] Figure 1 This is a graph showing the results of the strain antagonism experiment; Figure 2 The graph shows the results of the combined phosphorus solubility determination of two fungal strains. Figure 3 Figure showing the sporulation results of Penicillium desertis with different inoculation amounts; Figure 4 Figure 1 shows the sporulation results of *Penicillium desertii* and its composite strain with different material-to-liquid ratios.
[0014] desert penicillium ( Penicillium desertorumIt was deposited at the China General Microbiological Culture Collection Center on January 9, 2025, with accession number CGMCC NO: 41748; Penicillium fimbriae ( Penicillium steckii It was deposited on January 8, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO: 41749. Detailed Implementation
[0015] Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without inventive effort are within the scope of protection of this invention. The invention will now be described with reference to specific embodiments. It should be noted that these embodiments are merely descriptive and do not limit the invention in any way.
[0016] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.
[0017] The culture medium formulations described in the following examples are as follows: PDB medium (1L formulation): Peel 200.0 g of potatoes, cut into small pieces, add 1000 mL of water and boil for 30 min. Filter with double-layer gauze, take the filtrate and add 20 g of glucose, heat over low heat and stir constantly with a glass rod, and add distilled water to make up to 1000 mL, and adjust the pH to 5.
[0018] PDA medium: Add 18 g of agar per liter of PDA medium.
[0019] NBRIP liquid medium (g / L): glucose 10.0 g, Ca3(PO4)2 5.0 g, MgCl2·6H2O 5.0 g, MgSO4·7H2O 0.25 g, KCl 0.2 g, (NH4)2SO4 0.1 g, pH 7.2.
[0020] NBRIP solid medium: Add 18 g of agar per liter of NBRIP liquid medium.
[0021] The basic properties of the purchased mushroom substrate are as follows: The pH was 8.77, and the total phosphorus was 1.43 × 10⁻⁶. 4 mg / kg, total nitrogen is 2.24×10 mg / kg. 4 mg / kg, total potassium is 2.07×10 mg / kg. 4 mg / kg, organic matter is 641 g / kg.
[0022] Example 1 Strain antagonism experiment Two phosphate-solubilizing fungi were inoculated onto PDA medium using the streak-cross method and cultured at 30 °C for 5 days. Fungal growth at the crossover point was observed every 24 hours. The results of the combined culture on day 5 are shown below. Figure 1 As shown in the diagram, the strains at the intersection of the doodled lines are growing well and show no antagonistic reaction, and can be mixed and cultured to prepare phosphate-solubilizing agents.
[0023] Example 2 Activated *Penicillium desertii* and *Penicillium fimbriatum* were inoculated onto PDA medium and incubated at 30°C for 7 days until spores matured. After maturation, the spores were washed off the plates with sterile water and counted using a hemocytometer. The spore concentration was then diluted to obtain a concentration of 1 × 10⁻⁶ spores. 8 Spore suspensions of two fungal strains were prepared at a 1:1 ratio and inoculated into NBRIP medium at different inoculation ratios (1%, 3%, 5%, 7%, 9%). The mixtures were then incubated on a shaker at 170 r / min and 30 ℃. Every 24 h, 200 μL of the test solution was aspirated into plastic microplates, and the absorbance was measured at OD720 nm using a microplate reader. The available phosphorus content of the test solution was calculated. Results are shown below. Figure 2 As shown ( Figure 2 A, B, C, D, E, and F in the table represent the combined phosphorus solubility of the two fungi on days 3, 5, 7, 9, 11, and 13, respectively. The phosphorus solubility in each treatment group generally showed an initial increase followed by a decrease, with the highest phosphorus solubility in the 9% inoculum treatment group reaching 984.43 mg / L on day 9. Figure 2 D).
[0024] Example 3 The prepared spore suspension of *Penicillium desertii* (1×10⁻⁶) 8 Spores / mL were inoculated into mycelium bran (1 g mycelium bran plus 1 mL distilled water) at different inoculum amounts, with three treatment groups: 7%, 9%, and 11%. The culture was incubated at 28 ℃ and 150 r / min for 3 days. The culture medium was filtered through four layers of gauze, and spore yield was recorded using a hemocytometer. The experiment was repeated three times. Results are shown below. Figure 3 As shown, the number of spores of *Penicillium desertii* increased with the increase in inoculation amount. The 9% inoculation amount treatment group was not significantly different from the other two treatment groups. Based on Example 2 and considering the economic benefits of practical application in farmland, 9% was determined as the fungal inoculation amount for subsequent experiments.
[0025] Based on the determined inoculum size, three different material-to-liquid ratio treatment groups were set up: 1:3, 1:5, and 1:7 (3 g of crushed inoculum was added to 9 mL, 15 mL, and 21 mL of distilled water, respectively). Sterilization, inoculation, cultivation, and counting were then performed. In 50 mL Erlenmeyer flasks, inoculum and water were added according to the ratio and mixed thoroughly. The flask openings were sealed with sealing film and autoclaved for 30 min. After cooling to room temperature, 1×10⁻⁶ inoculum was added to each Erlenmeyer flask at a 9% inoculum ratio on a sterile operating table. 8 A mixed fungal spore suspension (Penicillium desertii and Penicillium fimbriae in a 1:1 ratio) was prepared at 28 °C and 150 r / min for 3 days. The culture medium was then filtered through four layers of gauze, and the cells were counted using a hemocytometer. The effect of different substrate-to-solvent ratios on the sporulation yield of Penicillium desertii was investigated. Figure 4 As shown in Figure A, the number of spores in each group was significantly increased compared to those that were not sieved, therefore, the substrate was pretreated by sieving in subsequent experiments. The spore production of the compound strain is shown in Figure A. Figure 4 As shown in Figure B, the highest sporulation rate of *Penicillium desertii* reached 5.56 × 10⁻³ when the material-to-liquid ratio was 1:3. 10 The sporulation rate of the compound bacterial strain reached 6.17 × 10⁶ cells / mL. 10 per mL.
[0026] Example 4 Add 90 mL of distilled water to 30 g of sterilized carrier substrate at a substrate-to-liquid ratio of 1:3, and add fungal spore suspension (1×10⁻⁶) at a 9% inoculation rate. 8 Solid phosphate-solubilizing fungal inoculum was prepared (spores / mL) and its technical indicators were determined.
[0027] Weigh 15 g of solid bacterial agent into a 50 mL beaker, add an appropriate amount of distilled water, stir evenly with a glass rod, let stand for 30 min, and measure the pH of the suspension using a pH meter. Perform three repeated experiments.
[0028] Place two test sieves with apertures of 4.75 mm and 1.0 mm stacked together on a base plate. Pour 50 g of solid bacterial agent sample into the 4.75 mm test sieve and sieve it. Weigh the sample from the 1.0 mm test sieve. The formula for calculating particle fineness is: In the formula: g is the mass fraction of the inoculum (%); m1 and m2 are the mass of the sample on the 1.0 mm experimental sieve and the mass of the solid inoculum sample, respectively, in grams (g). Three replicate experiments were conducted.
[0029] Place the empty aluminum box in a drying oven at 105 ℃ for 30 min to ensure there is no moisture inside. Remove the box and cool it to room temperature. Weigh and record its mass. Weigh 20 g of the sample from a 1.0 mm test sieve into the aluminum box. Weigh the total mass and place the box in the drying oven at 105 ℃ for 5 h to ensure complete evaporation of moisture from the sample. Remove the box and cool it to room temperature for 20 min. Weigh the total mass of the box and sample again. The moisture content formula is: In the formula: w is the moisture content of the sample (%); m1 is the mass of the sample and aluminum box, in grams (g); m2 is the mass of the dried sample and aluminum box, in grams (g); m0 is the mass of the empty aluminum box, in grams (g). Three repeated experiments were conducted.
[0030] The effective viable count was determined using the plate count method. The specific steps are as follows: Weigh 10 g of solid sample and add it to 100 mL of sterile water (for liquid samples, add 10 mL to 90 mL of sterile water), let stand for 20 min, then shake thoroughly at 200 r / min for 30 min to obtain 10 g of viable bacteria. -1 Bacterial suspension, repeated steps for serial dilution to obtain 10 -2 10 -3 10 -4 10 -5 10 -6 and 10 -7 Bacterial suspension. Take 0.1 mL of three consecutive appropriately diluted bacterial suspensions and spread them evenly on PDA medium. Each dilution is repeated three times, with sterile water as a blank control. Incubate the plates at 28 °C for 5 days. Plates showing 20–300 colonies are used as the counting standard. The calculation formula is as follows: Where: n m The effective viable count is expressed as 100 million per gram (100 million / g); x is the average colony count, expressed as cells; k is the dilution factor; V1 is the basal liquid volume, expressed as milliliters (mL); m0 is the sample volume, expressed as grams (g); V2 is the volume of bacterial suspension added, expressed as milliliters (mL); n v V0 represents the effective viable count per volume, expressed in billions per milliliter (billions / mL); V0 represents the sample volume, expressed in milliliters (mL).
[0031] The technical specifications are shown in Table 1: pH 8.18, fineness 80.0, moisture content 19.32%, and viable bacteria count 2.2 × 10⁻⁶. 8 The CFU / g values all meet the standards for agricultural microbial inoculants (GB20287-2006).
[0032] Table 1. Comparison of Technical Indicators of Compound Microbial Agent with National Standards The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. A soil phosphorus-solubilizing solid bacterial fertilizer, characterized by, including Penicillium desertorum (ATCC 9645) Penicillium steckii ) and Penicillium variabile (ATCC 10408) Penicillium steckii ). The desert penicillium fungus (Penicillium desertsom Penicillium steckii ) was deposited at China General Microbiological Culture Collection Center on January 9, 2025, and the deposit number is CGMCC NO: 41748. The Penicillium fissure ( Penicillium steckii It was deposited on January 8, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO: 41749.
2. The soil phosphorus-solubilizing solid bacterial manure of claim 1, characterized in that, The ratio of the Penicillium desertorum to the Penicillium steckii is 1:
1.
3. The soil phosphorus-soluble solid bacterial fertilizer according to claim 1, characterized in that, The soil phosphorus dissolving solid bacterial fertilizer can reach a phosphorus dissolving amount of 984.43 mg / L.
4. The soil phosphorus-soluble solid bacterial fertilizer according to claim 1, characterized in that, The soil phosphorus dissolving solid bacterial fertilizer further comprises agricultural waste bacterial chaff.
5. The soil phosphorus-soluble solid bacterial fertilizer according to claim 4, characterized in that, The agricultural waste bacterial chaff is mixed with distilled water, and the solid-liquid ratio of the agricultural waste bacterial chaff to the distilled water is 1:
3.
6. The soil phosphorus-solidifying solid bacterial fertilizer according to claim 3, characterized in that, The inoculation amount of the bacterial liquid is 9wt%.
7. The soil phosphorus-solubilizing solid bacterial manure according to claim 1, characterized by, The soil-soluble phosphorus solid bacterial fertilizer has a pH of 8.18, a fineness of 80.0, a moisture content of 19.32%, and a viable bacterial count of 2.2×10 8 CFU / g.