Rice whole-metabolism organic fertilizer, preparation method and application thereof

Abstract

The application discloses a kind of rice full element biological organic fertilizer and its preparation method and application.The rice full element biological organic fertilizer of the application includes active functional bacteria component, organic fertilizer component, amino acid component and inorganic compound fertilizer component, the active functional bacteria component contains the bacillus velezensis of promoting growth and reducing cadmium, the bacillus amyloliquefaciens of disease resistance and growth promotion and rice disease resistance and growth promotion endophytic streptomyces;The organic fertilizer component contains rotten sawdust, mushroom residue, general bran, chicken manure and pig manure.The application also includes the preparation method of the rice full element biological organic fertilizer.The fertilizer of the application has the advantages of organic fertilizer, inorganic fertilizer, microbial fertilizer and amino acid fertilizer, can improve fertilizer utilization, improve soil physical and chemical properties, nutritional status and biological activity, promote the growth and reproduction of rhizosphere microorganisms of rice, enhance the disease resistance level of rice, provide comprehensive and balanced nutrition for the growth of rice, and significantly improve the yield and quality of rice.

Description

Technical Field

[0001] This invention belongs to the field of microbial fertilizer and its preparation technology, and relates to a rice all-element biological organic fertilizer, its preparation method and application. Background Technology

[0002] my country is a major agricultural country, producing 21% of the world's food using only 8% of the world's arable land. However, it also consumes 35% of the world's chemical fertilizers, with fertilizer usage per unit area three times the world average. Long-term, continuous, and excessive application of chemical fertilizers has led to a continuous decrease in soil organic matter, resulting in declining soil fertility due to salinization, acidification, and compaction. This also pollutes water sources, damages the ecological environment, and harms human and animal health. Furthermore, it has caused previously high-yield fields to trend towards becoming "infertile" and low-yield fields.

[0003] In response to the various problems currently existing in my country's soil, such as declining basic soil fertility, resource depletion, and environmental pollution, priority should be given to developing the green fertilizer industry.

[0004] Increasing the application of organic matter and enhancing the diversity of soil microbial communities are the main ways to improve soil fertility. Bio-organic fertilizers have gained widespread attention due to their characteristics such as regulating plant growth, increasing crop yield, improving crop quality, reducing chemical fertilizer use, improving soil quality, and protecting the ecological environment, making them an irreplaceable agricultural input in the development of green ecological agriculture. However, while single organic fertilizers can enrich the soil, their nutrient content is low and their release is slow, resulting in limited effects on increasing the yield of current season crops. Furthermore, existing bio-organic fertilizers often contain microorganisms with limited function and low efficacy, leading to unstable effects on current season crops.

[0005] All-element bio-organic fertilizer refers to a new type of compound fertilizer that integrates compost, chemical fertilizer, bio-fertilizer, and amino acid fertilizer. Its functions include promoting the growth of seasonal crops, significantly inhibiting soil acidification, effectively regulating the soil microbial flora, and continuously improving basic soil fertility. Applying all-element bio-organic fertilizer can simultaneously increase soil organic matter content and soil biological activity, maintaining a good balance among soil mineralizing microorganisms, soil nitrifying bacteria, soil nitrogen-fixing bacteria, and soil phosphorus- and potassium-solubilizing bacteria, thus continuously and stably outputting soil fertility. Summary of the Invention

[0006] The primary objective of this invention is to provide a method for preparing a complete bio-organic fertilizer for rice. The rice complete bio-organic fertilizer prepared by this method combines the advantages of organic fertilizer, inorganic fertilizer, microbial fertilizer, and amino acid fertilizer. It can improve fertilizer utilization, enhance soil physicochemical properties, nutrient status, and biological activity, promote the growth and reproduction of rhizosphere microorganisms in rice, strengthen rice's disease resistance, provide comprehensive and balanced nutrition for rice growth, and significantly improve rice yield and quality.

[0007] A method for preparing a complete bio-organic fertilizer for rice includes the following steps:

[0008] (1) First, liquid fermentation was carried out on Bacillus belye 20-10 and Bacillus amyloliquefaciens CWJ2, and then the fermentation liquid was added to the decomposed organic fertilizer for solid fermentation.

[0009] (2) Solid-state fermentation of Streptomyces Ahn65;

[0010] (3) The fermentation products obtained in steps (1) and (2) are then mixed with amino acids and inorganic compound fertilizer to obtain rice all-element biological organic fertilizer.

[0011] The preservation numbers of Bacillus belyssus 20-10, Bacillus amyloliquefaciens CWJ2, and Streptomyces Ahn65 are CCTCC NO:M 20221980, CCTCC NO:M 20221983, and CCTCC NO:M20221981, respectively.

[0012] Step (1) specifically includes:

[0013] a. Activation of bacterial strains: Bacillus belye 20-10 and Bacillus amyloliquefaciens CWJ2 were separately inoculated onto solid LB medium to activate the bacterial strains, and then separately inoculated into LB liquid medium for culture to prepare seed culture;

[0014] b. Fermentation culture: Inoculate the seed liquid separately into the fermenter at an inoculation rate of 0.5% to 2% by volume, and introduce sterile air for fermentation culture;

[0015] When the fermentation broth of both bacteria contains more than 5 billion live spores per milliliter and the spore ratio is more than 90%, the broth is placed in a tank with a pH of 7.0-8.0 and no contamination by other bacteria.

[0016] c. Adjust the carbon-nitrogen ratio of the decomposed organic fertilizer to 20:1 to 30:1, and the moisture weight ratio to 40 to 60%. Inoculate with the fermentation liquid of Bacillus belye and Bacillus amyloliquefaciens at a ratio of 0.5 to 2%, mix evenly, and carry out natural composting fermentation.

[0017] Furthermore, step (1) includes more specifically:

[0018] a. Activation of bacterial strains: Bacillus belye 20-10 and Bacillus amyloliquefaciens CWJ2 were inoculated on solid LB medium and cultured at 30°C for 16-30 h to activate the bacterial strains. Then, the bacterial cells were inoculated into 100 mL of LB liquid medium with a sterile loop and cultured at 30°C and 180 rpm for 16-24 h to prepare the seed culture.

[0019] b. Fermentation culture: First, sterilize the fermenter, fill it with culture medium and then sterilize it again. Maintain pressure and cool down to 30℃. Inoculate the seed tank fermentation liquid into the fermenter at an inoculation rate of 0.5-2% and introduce sterile air. Culture at 30℃ for 60-72 hours with an aeration rate of 1.5 Vols / vol·min and an oxygen content of 35% by volume.

[0020] Fermentation medium formula: maltose 2%, starch 1%, yeast extract 1.25%, NaCl 0.1%, ammonium sulfate 0.3%, calcium carbonate 0.1%, balance water, initial pH 7.2-7.5;

[0021] c. Adjust the carbon-nitrogen ratio of the decomposed organic fertilizer to 20:1 to 30:1, and the moisture content to 40 to 60% by weight. Inoculate with Bacillus vesiculosus and Bacillus amyloliquefaciens fermentation liquid at an inoculation rate of 0.5 to 2%, mix evenly, and carry out natural composting fermentation. When the temperature reaches above 50℃, start turning the pile, turning it 2 to 3 times a day. Continue until the organic fertilizer germination index is ≥90% to complete the composting fermentation.

[0022] The decomposed organic fertilizer is obtained by composting sawdust, mushroom residue, rice bran, chicken manure, and pig manure in a weight ratio of 0.5:0.5:(2-3):(2-3):4.

[0023] Further optimization involves step c, adjusting the carbon-to-nitrogen ratio of the decomposed organic fertilizer to 25:1, adjusting the moisture content to 40%, and inoculating the fermentation broth of Bacillus belye and Bacillus amyloliquefaciens at an inoculation rate of 0.5%, mixing evenly, and then carrying out natural composting fermentation.

[0024] Step (2) specifically includes:

[0025] a. Activation of microbial strains

[0026] Streptomyces Ahn65 was activated by inoculating it into ISP2 solid medium, and then transferred to ISP2 liquid medium to prepare seed culture.

[0027] b. Solid-state fermentation

[0028] Inoculate sterile wheat bran with the seed culture of Streptomyces Ahn65 at a mass ratio of 1-5%, and let it stand in the culture until the spore content of Streptomyces reaches more than 5 billion / g.

[0029] c. Dry the solid fermentation product of Streptomyces Ahn65 to prepare Streptomyces powder.

[0030] Furthermore, step (2) includes more specifically:

[0031] a. Activation of microbial strains

[0032] Streptomyces Ahn65 was activated by inoculating it into ISP2 solid medium, and then transferred to 50 mL of ISP2 culture medium. The culture was carried out at 28℃ and 180 rpm for 1-2 days to prepare seed culture.

[0033] b. Solid-state fermentation

[0034] Inoculate sterile wheat bran with the culture medium of Streptomyces Ahn65 at a mass ratio of 1-5%, add water to make up the water mass ratio of 40-60%, and incubate at 28℃ for 10-20 days. Stop the culture when the spore content of Streptomyces reaches more than 5 billion / g.

[0035] c. The solid fermentation product of Streptomyces Ahn65 was dried in a 40℃ incubator for 24 hours to prepare Streptomyces mycelium powder.

[0036] Further, Streptomyces powder is added to the product of step (1) at a mass ratio of 1-2%, mixed evenly, and then dried at low temperature until the moisture content is ≤30%, thus obtaining a bio-organic fertilizer rich in three functional microorganisms: Bacillus belye, Bacillus amyloliquefaciens, and Streptomyces, with each microorganism having an effective viable count of not less than 200 million / g.

[0037] The prepared bio-organic fertilizer is mixed with 5-8% amino acid raw powder and inorganic compound fertilizer by mass percentage, and then mixed evenly to make a complete bio-organic fertilizer. The complete bio-organic fertilizer has an effective live bacteria count of 6-8 billion / g, an organic matter content of 30-55%, a total NPK nutrient content of 8-11%, a moisture content of ≤30%, a total amino acid content of 5-10%, a free amino acid content of 0.5-5%, a chloride ion content of ≤2.0%, and a pH of 6.0-7.0.

[0038] This invention also provides a complete bio-organic fertilizer for rice, which is prepared by the above method.

[0039] This invention also provides the application of the aforementioned rice whole-element biological organic fertilizer, which promotes rice growth, improves soil nutrition, and enhances rice disease resistance.

[0040] Compared with existing technologies such as the patent "A compound microbial agent and its application in promoting growth, resisting disease and reducing cadmium in rice (application number CN2023110559849)", the present invention has the following beneficial effects:

[0041] (1) The whole element biological organic fertilizer of the present invention has a more significant effect on increasing rice yield. When applied at a rate of 150 kg / mu, it can increase rice yield by 41.59%-42.71%, which is much higher than the yield increase effect of the compound microbial agent in patent CN2023110559849 (19.35%).

[0042] (2) The highest control efficacy of the whole element biological organic fertilizer of the present invention against rice leaf blast is 66.08%, which is higher than the control efficacy of the compound microbial agent against rice leaf blast (56.74%) in CN2023110559849.

[0043] (3) The whole element biological organic fertilizer of the present invention not only promotes rice growth and improves rice resistance to rice blast, but also effectively enhances soil fertility and microbial diversity. The soil organic matter and total nitrogen, available nitrogen, available phosphorus and available potassium contents increased by 8.82%, 13.46%, 7.80%, 197.87% and 39.23% respectively compared with the blank, and the total number of soil microorganisms was greater and the microbial abundance was greater. The Streptomyces Ahn65 used in this invention is classified as Streptomyces sp., with accession number CCTCCNO:M 20221981, accession date of December 19, 2022, and deposited at the China Center for Type Culture Collection (CCTCC), located at No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province, within the campus of Wuhan University. The taxonomic name of Bacillus velezensis 20-10 is Bacillus velezensis, with accession number CCTCCNO:M20221980, deposit date of December 19, 2022, and depositary institution China Center for Type Culture Collection (CCTCC), located at No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province, within the campus of Wuhan University. The taxonomic name of Bacillus amyloliquefaciens CWJ2 is Bacillus amyloliquefaciens, with accession number CCTCCNO:M 20221983, deposit date of December 19, 2022, and deposited at the China Center for Type Culture Collection (CCTCC), located at Wuhan University, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province. Detailed Implementation

[0044] The present invention will be further described in detail below with reference to the embodiments.

[0045] Example 1: Preparation of All-Element Bio-Organic Fertilizer

[0046] (1) Preparation of active functional bacteria:

[0047] First, Bacillus belye and Bacillus amyloliquefaciens were fermented separately in fermenters. The operating steps and process conditions were as follows:

[0048] a. Activation of microbial strains

[0049] Bacillus belye 20-10 and Bacillus amyloliquefaciens CWJ2 were inoculated onto solid LB medium and cultured at 30°C for 24 hours to activate the bacterial strains. Then, a spoonful of bacterial cells was scooped out with a sterile inoculation loop and inoculated into 100 mL of LB liquid medium (in a 500 mL Erlenmeyer flask) sterilized at 121°C for 30 min. The culture was then carried out at 30°C and 180 rpm for 24 hours to prepare the seed culture.

[0050] b. Fermentation tank cultivation

[0051] First, clean and sterilize the fermenter, fill it with fermentation medium and sterilize it again, maintain pressure and cool it down to 30°C, inoculate the seed liquid into the fermenter at an inoculation rate of 1% by volume, and introduce sterile air.

[0052] The sterilization process employs high-pressure steam sterilization, specifically at 121°C and a pressure of 0.4 kg / cm². 2 Sterilize for 30 min under the specified conditions; after inoculation in the fermenter, incubate at 30℃ for 72 h with an aeration rate of 1.5 Vols / vol·min and an oxygen content of 35% by volume.

[0053] LB medium formulation: 1% tryptone, 5% yeast extract, 0.5% sodium chloride, water, pH 7.2, with 1.8-2.0% agar added to the solid medium;

[0054] Fermentation medium formula: maltose 2%, starch 1%, yeast extract 1.25%, NaCl 0.1%, ammonium sulfate 0.3%, calcium carbonate 0.1%, balance water, initial pH 7.2-7.5;

[0055] The percentage content of all components in the culture medium formulation is by weight.

[0056] Fermentation broth requirements: The fermentation broth should contain more than 5 billion live spores per milliliter, with a spore ratio of more than 90%, before being placed in the tank. The pH value should be 7.0-8.0, and there should be no contamination by other microorganisms.

[0057] c. Adjust the carbon-to-nitrogen ratio of the decomposed organic fertilizer to 25:1 and the moisture content to 40% by weight. Inoculate with the fermentation broth of Bacillus belye and Bacillus amyloliquefaciens at a ratio of 0.5%, mix thoroughly, and allow for natural composting. When the temperature reaches above 50℃, begin turning the compost pile 2-3 times daily until the organic fertilizer germination index is ≥90%, thus completing the composting process.

[0058] The aforementioned composted organic fertilizer is obtained by composting sawdust, mushroom residue, rice bran, chicken manure, and pig manure in a weight ratio of 0.5:0.5:2:3:4.

[0059] (2) Preparation of active functional Streptomyces preparations:

[0060] a. Activation of microbial strains

[0061] Streptomyces Ahn65 was inoculated into ISP2 solid medium for activation, and then transferred to 50 mL of ISP2 culture medium. The culture was carried out at 28 °C and 180 rpm for 1-2 days to prepare seed culture.

[0062] b. Solid-state fermentation

[0063] The culture medium of Streptomyces Ahn65 was inoculated into sterile wheat bran at a volume ratio of 1%, and water was added until the water volume ratio was 40%. The mixture was then incubated at 28°C for 20 days. The culture was stopped when the spore content of Streptomyces reached more than 5 billion / g.

[0064] c. Preparation of Streptomyces preparations

[0065] The solid-state fermentation product of Streptomyces Ahn65 was dried in an incubator at 40℃ for about 24 hours to prepare Streptomyces mycelium powder, which was then sealed and stored in bags. Plate dilution and counting were performed, and the effective viable count reached 200-500 billion / g.

[0066] (3) Preparation of bio-organic fertilizer:

[0067] The Streptomyces preparation is added at a mass ratio of 1% to the product of the secondary fermentation of the functional bacteria prepared in step (1), which is the bio-organic fertilizer. After mixing evenly, it is transferred to a low-temperature dryer for drying until the moisture content is ≤30%. This yields a bio-organic fertilizer rich in three functional microorganisms: Bacillus belye, Bacillus amyloliquefaciens, and Streptomyces. The effective viable count of each microorganism is not less than 200 million / g.

[0068] (4) Preparation of complete elemental bio-organic fertilizer:

[0069] The amino acid raw powder was purchased from Shandong Weifang Lvlong Biotechnology Co., Ltd., and contains 80% plant-derived amino acids. The compound fertilizer was purchased from Shandong Stanley Fertilizer Co., Ltd., with a total nutrient content (N+P2O5+K2O) of 50%. The prepared bio-organic fertilizer was fed into a silo, and 6.9% by weight of amino acid raw powder and compound fertilizer were added. After mixing evenly, the mixture was packaged to produce a complete bio-organic fertilizer.

[0070] The Quanyuan biological organic fertilizer contains 6-8 billion live bacteria per gram, 30-55% organic matter, 8-11% total NPK nutrients, ≤30% moisture, 5-10% total amino acids, 0.5-5% free amino acids, ≤2.0% chloride ion content, and a pH of 6.0-7.0.

[0071] Example 2: An exploratory experiment on organic fertilizer components:

[0072] 1. Preparation of organic fertilizer components:

[0073] Preparation of organic fertilizer 1 with different degrees of decomposition: Chicken manure and pig manure treated at 85℃ for 2 days were mixed with crushed sawdust, mushroom residue and rice bran in a weight ratio of 3:4:0.5:0.5:2. The mixture was then transferred to a large solid composting tank. The carbon-nitrogen ratio was adjusted to 30:1 and the moisture content was adjusted to 50%. An organic material composting agent was added and composting was carried out. When the pile temperature reached above 60℃, the pile was turned and aerated. The pile was turned 3 times a day. The compost fermented for 15 days. A portion was taken out as incompletely decomposed organic fertilizer 1 and composted for about 20 days. During this period, a small amount of water could be added appropriately until the organic fertilizer germination index was ≥90% and the compost was fully decomposed. The composting was then stopped to obtain decomposed organic fertilizer 1.

[0074] Preparation of organic fertilizer 2 with different degrees of decomposition: Pig manure and rice bran treated at 85℃ for 2 days were screened, crushed and mixed evenly in a weight ratio of 8:2. The mixture was then transferred to a large solid composting tank, the carbon-nitrogen ratio was adjusted to 30:1 and the moisture content was adjusted to 50%. Organic material composting agent was added and composting was carried out. When the pile temperature reached above 60℃, the pile was turned and aerated. The pile was turned 3 times a day. The compost fermented for 15 days. A portion was taken out as incompletely decomposed organic fertilizer 2. The remainder was composted for 35 days until the organic fertilizer germination index was ≥90% and the compost was complete. Composting was then stopped to obtain decomposed organic fertilizer 2.

[0075] Organic fertilizer 3: A certain brand of organic fertilizer purchased from the Mawangdui Farmers Market in Changsha.

[0076] 2. Secondary fermentation by functional bacteria:

[0077] 1) Exploring the formula for organic fertilizer used in secondary fermentation:

[0078] The conditions and levels of secondary fermentation of different types of functional microorganisms, including Bacillus belyssus, Bacillus amyloliquefaciens and Streptomyces, were explored under different raw material ratios, different degrees of composting and different sources of organic fertilizer.

[0079] Fermentation broths of *Bacillus belye*, *Bacillus amyloliquefaciens*, and spore suspensions of *Streptomyces* were inoculated into five different organic fertilizers at a 1% inoculum. Water was added until the water-to-nitrogen ratio reached 40% by volume, and the carbon-to-nitrogen ratio was adjusted to 25:1. After thorough mixing, secondary composting fermentation was carried out. A control group without microbial inoculation was also included. When the temperature reached above 50℃, the compost was turned 2-3 times daily. Samples were taken on the third day, diluted using the plate dilution method, and then divided into 10 dilution gradients. -5 -10 -7 The samples were plated, and the number of colonies in different dilution gradients was counted. The average number of target microorganisms in the samples was calculated based on the dilution factor.

[0080] The results are shown in Table 1. Both *Bacillus belyceae* and *Bacillus amyloliquefaciens* showed the highest proliferation levels when using well-rotted organic fertilizer 1 as the secondary fermentation material, followed by well-rotted organic fertilizer 2. Secondary fermentation with unrotted organic fertilizer 1, unrotted organic fertilizer 2, and commercially available organic fertilizer resulted in low proliferation levels. This indicates that the degree of organic fertilizer decomposition is a key indicator for the secondary fermentation of functional bacteria. Furthermore, the raw material ratio and nutrient composition of the organic fertilizer also affect the secondary fermentation of functional bacteria. In this experiment, *Streptomyces* did not show effective proliferation in any of the organic fertilizers used for secondary fermentation, suggesting that *Streptomyces* may not be suitable for secondary fermentation in organic fertilizers.

[0081] Table 1. Level of secondary fermentation by microorganisms in different organic fertilizers (Unit: *10) 8 CFU / g

[0082]

[0083] 2) Exploration of secondary fermentation conditions

[0084] Based on the above conclusions, well-rotted organic fertilizer 1 was selected as the organic fertilizer for secondary fermentation, and the conditions for secondary fermentation were explored.

[0085] The moisture-to-volume weight ratio of the organic fertilizer was adjusted to 40%, 50%, and 60% respectively. *Bacillus bellis* and *Bacillus amyloliquefaciens* were inoculated at a 1% inoculum rate, mixed thoroughly, and then subjected to solid-state fermentation at room temperature indoors. When the temperature reached above 50℃, the compost was turned 2-3 times daily. After 3 days of natural composting, the number of *Bacillus bellis* and *Bacillus amyloliquefaciens* bacteria in secondary fermentation was determined using the plate count method. The results are shown in Table 2. The number of *Bacillus bellis* and *Bacillus amyloliquefaciens* bacteria in secondary fermentation decreased with increasing moisture content; a moisture content of 40% was optimal.

[0086] Table 2. Effect of different moisture contents on the number of bacteria in the secondary fermentation of functional bacteria. Unit: *10 8 CFU / g

[0087]

[0088] The carbon-to-nitrogen ratio of the decomposed organic fertilizer 1 was adjusted to 25-30:1, and the moisture content (by volume) was 40%. *Bacillus bellis* and *Bacillus amyloliquefaciens* were inoculated at inoculum rates of 0.5%, 1%, and 2%, respectively. Once the temperature reached above 50℃, the compost was turned 2-3 times daily. After 3 days of natural composting, the number of *Bacillus bellis* and *Bacillus amyloliquefaciens* bacteria in secondary fermentation was determined using the plate count method. The results are shown in Table 3. The number of *Bacillus bellis* and *Bacillus amyloliquefaciens* bacteria in secondary fermentation was highest with an inoculum rate of 2%, but considering cost and convenience, an inoculum rate of 0.5% was preferable.

[0089] Table 3. Effect of different inoculum sizes on the number of functional bacteria in secondary fermentation. Unit: *10 8 CFU / g

[0090]

[0091] The carbon-nitrogen ratio of the decomposed organic fertilizer 1 was adjusted to 20:1, 25:1, and 30:1, and the moisture content (volume weight ratio) was adjusted to 40%. *Bacillus bellis* and *Bacillus amyloliquefaciens* were inoculated at a ratio of 0.5%, mixed thoroughly, and allowed to undergo natural composting fermentation. When the temperature reached above 50℃, the compost was turned 2-3 times daily. After 3 days of natural composting fermentation, the number of *Bacillus bellis* and *Bacillus amyloliquefaciens* bacteria in secondary fermentation was determined using the plate count method. The results are shown in Table 4. The optimal carbon-nitrogen ratio for secondary fermentation by *Bacillus bellis* and *Bacillus amyloliquefaciens* was 25:1. At this point, the highest fermentation level of *Bacillus bellis* was 5.932 billion / g, while the highest fermentation level of *Bacillus amyloliquefaciens* reached 1.585 billion / g. This significantly reduced costs for the large-scale propagation and application of functional microorganisms.

[0092] Table 4. Effects of different carbon-to-nitrogen ratios on the number of bacteria in secondary fermentation of functional microorganisms. Unit: *10 8 CFU / g

[0093]

[0094] Example 3: Field trial of rice using all-element biological organic fertilizer:

[0095] (1) Experimental Design:

[0096] The field trial was conducted in Yiyang City, Hunan Province, from July to November 2023, covering an area of ​​5 mu (approximately 0.33 hectares). The soil texture was clay loam, and the soil type was red-yellow mud. The previous crop was rapeseed, followed by purple clover green manure. The basic physicochemical properties of the soil were: pH 5.24, organic matter 38.93 g / kg, total nitrogen 2.22 g / kg, available phosphorus 7.97 mg / kg, available potassium 176.17 mg / kg, and Cd 0.32 mg / kg.

[0097] The experimental field consisted of 4 treatments, with 3 plots per treatment, for a total of 12 plots. Each plot had an area of ​​20m². 2 The experimental field was surrounded by a 3-meter-wide protective row. The plots were randomly arranged, and the entire field ridge above the plowed layer was covered with agricultural film to prevent cross-contamination of fertilizer and water. Four treatments were implemented, with identical field management and pest and disease control practices.

[0098] Treatment 1: Apply a combination of whole-element bio-organic fertilizer and reduced chemical fertilizer, i.e., apply 80% of the conventional fertilizer before rice transplanting, i.e., apply 16 kg / 667 m² of compound fertilizer. 2As a base fertilizer, apply 150 kg / 667 m² of complete elemental bio-organic fertilizer one week after rice transplanting. 2 .

[0099] Treatment 2: Inactivated all-element biological organic fertilizer + reduced chemical fertilizer application, i.e., applying 80% of the conventional fertilizer before rice transplanting, i.e., applying 16 kg / 667 m² of compound fertilizer. 2 As a base fertilizer, apply 150 kg / 667 m² of inactivated all-element bio-organic fertilizer one week after rice transplanting. 2 .

[0100] Treatment 3: Conventional fertilization, applying compound fertilizer at 20 kg / 667 m² according to local recommendations. 2 .

[0101] Treatment 4: Blank control (CK) - no fertilizer applied.

[0102] (2) Experimental results:

[0103] The application of the all-element bio-organic fertilizer significantly promoted the growth and tillering of rice. Compared with the control group and conventional fertilization, the plant height increased by 25.10% and 14.96%, respectively, while the number of tillers increased by 75.78% and 57.39%, respectively. The wet weight and dry weight of the plots increased by 41.59% and 42.71% compared with the control group, respectively, and by 18.83% and 17.74% compared with conventional fertilization. This is the first all-element bio-organic fertilizer used for rice, and its yield increase level is far higher than that of commercially available bio-organic fertilizers and organic fertilizers, as shown in Tables 5 and 6.

[0104] Table 5. Effects of different treatments on economic traits of rice.

[0105]

[0106]

[0107] Table 6. Effects of different treatments on rice yield

[0108]

[0109] Meanwhile, this all-element bio-organic fertilizer can also improve the disease resistance of rice, with a disease index control efficacy of up to 66.08% against rice leaf blast (Table 7), and can effectively improve soil nutrient indicators such as nitrogen, phosphorus, and potassium, as well as soil organic matter content (Table 8). The treatment with the all-element bio-organic fertilizer significantly improved the abundance and diversity of soil microorganisms, with indicators representing the number of soil microbial species (OTU number), total species index (ace index, Chao index), and species biodiversity (shannon) all significantly higher than the control (Table 9).

[0110] Table 7. Effects of different treatments on late-season rice leaf blast.

[0111]

[0112] Table 8. Effects of different treatments on nutrient content in the rhizosphere soil of late-season rice.

[0113]

[0114] Table 9. Effects of different treatments on rhizosphere microbial diversity

[0115]

Claims

1. A method for preparing a complete bio-organic fertilizer for rice, characterized in that, Includes the following steps: (1) First, liquid fermentation was carried out on Bacillus belye 20-10 and Bacillus amyloliquefaciens CWJ2, and then the fermentation liquid was added to the decomposed organic fertilizer for solid fermentation. (2) Streptomyces Ahn65 was fermented in solid state to obtain Streptomyces powder; (3) Mix the solid fermentation product of step (1) and the Streptomyces powder obtained in step (2), as well as amino acids and inorganic compound fertilizer to obtain rice whole element biological organic fertilizer; The preservation numbers of Bacillus belyssus 20-10, Bacillus amyloliquefaciens CWJ2 and Streptomyces Ahn65 are CCTCC NO:M 20221980, CCTCC NO:M 20221983 and CCTCC NO:M 20221981, respectively. Step (1) includes: a. Activation of bacterial strains: Bacillus belye 20-10 and Bacillus amyloliquefaciens CWJ2 were inoculated onto solid LB medium to activate the bacterial strains. Then, the bacterial cells were inoculated into LB liquid medium using a sterile loop and cultured at 30°C and 180 rpm for 16-24 h to prepare seed culture. b. Fermentation culture: First, sterilize the fermenter, fill it with culture medium and then sterilize it again. Maintain pressure and cool down to 30°C. Inoculate the seed liquid into the fermenter at an inoculation rate of 0.5-2% and introduce sterile air. Culture at 30°C for 60-72 h with an aeration rate of 1.5 Vols / vol·min and an oxygen content of 35% by volume to obtain the fermentation broth. c. Adjust the carbon-to-nitrogen ratio of the decomposed organic fertilizer to 25:1 and the moisture content to 40% by weight. Inoculate with the fermentation broth of Bacillus belye and Bacillus amyloliquefaciens at a ratio of 0.5%, mix thoroughly, and allow for natural composting fermentation. When the temperature reaches above 50℃, begin turning the compost pile 2-3 times daily until the organic fertilizer germination index is ≥90%, thus completing the composting fermentation process. The aforementioned composted organic fertilizer is obtained by composting sawdust, mushroom residue, rice bran, chicken manure, and pig manure in a weight ratio of 0.5:0.5:2:3:

4. Step (2) includes: a. Activation of microbial strains Streptomyces Ahn65 was activated by inoculating it into ISP2 solid medium, and then transferred to ISP2 culture medium. The culture was carried out at 28℃ and 180 rpm for 1-2 days to prepare seed culture. b. Solid-state fermentation Inoculate sterile wheat bran with the seed culture of Streptomyces Ahn65 at a mass ratio of 1-5%, add water to make up to a moisture mass ratio of 40-60%, and incubate at 28℃ for 10-20 days. Stop the culture when the spore content of Streptomyces reaches more than 5 billion / g. c. The solid fermentation product of Streptomyces Ahn65 was dried in a 40℃ incubator for 24 hours to prepare Streptomyces mycelium powder; Rice-based whole-element biological organic fertilizer contains 6-8 billion live bacteria per gram, 30-55% organic matter, 8-11% total NPK nutrients, ≤30% moisture, 5-10% total amino acids, 0.5-5% free amino acids, ≤2.0% chloride ion content, and a pH of 6.0-7.

0.

2. The method for preparing rice-based complete bio-organic fertilizer according to claim 1, characterized in that, Step (1) Fermentation medium formula for liquid fermentation: maltose 2%, starch 1%, yeast extract 1.25%, NaCl 0.1%, ammonium sulfate 0.3%, calcium carbonate 0.1%, the remainder is water, and the initial pH value is 7.2 to 7.

5.

3. A rice-based complete bio-organic fertilizer, prepared by the method described in claim 1 or 2.

4. The application of the rice complete bio-organic fertilizer according to claim 3, characterized in that, The applications include: promoting rice growth, improving soil nutrition, and enhancing rice's disease resistance.

Images