Preparation method and application of continuous cropping resistant microbial fertilizer

Abstract

The invention discloses a preparation method and application of an anti-continuous cropping microbial fertilizer, and aims to solve the problems of aggravation of crop diseases and insect pests and reduction of yield and quality caused by continuous cropping planting. The preparation method comprises the following steps: activating and uniformly mixing bacillus subtilis and bacillus licheniformis, and adding an alpha-lactalbumin solution and a calcium chloride solution to prepare a bacillus aggregate; adding bentonite into the polytrehalose solution, and performing ultrasonic treatment on the mixture and the humic acid solution to prepare a modified polytrehalose-bentonite compound; uniformly mixing acremonium terricola, trichoderma harzianum and a bacillus aggregate, and adding a wall material solution to prepare a compound bacterium microcapsule preparation; and finally, mixing the microcapsules with the corn straw biochar, the chitosan oligosaccharide and the like to obtain a finished product. Through calcium ion induction and humic acid modified embedding, the strain stability and viable count are improved, soil organic matter and nitrogen phosphorus and potassium nutrients can be activated, pathogenic bacteria are inhibited, the crop rhizosphere environment is improved, the growth index and quality of red kidney beans and potatoes are effectively improved, and the method is suitable for continuous cropping resistant planting of crops.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of microbial fertilizer processing, and particularly relates to a preparation method of an anti-replanting microbial fertilizer and application thereof. BACKGROUND

[0002] Replanting easily causes replanting disorder, which leads to intensification of crop diseases and insect pests, and significant reduction of yield and quality. Replanting disorder belongs to soil-borne disease, which is mainly caused by unbalanced soil nutrition, lack of part of nutrient elements, change of soil physical and chemical properties, reduction of soil enzyme activity, and unbalanced microbial colony in crop rhizosphere due to multi-year replanting. The number of pathogenic microorganisms such as fungi, bacteria, nematodes and actinomycetes gradually increases, while the number of beneficial microorganisms decreases. In addition, continuous planting will cause crops to secrete some autotoxic substances (such as phenolic acids, organic acids and flavonoids) that inhibit crop growth, and these substances will continuously accumulate with the increase of planting years. Replanting disorder mainly harms the crop roots, hinders the roots from obtaining the nutrient substances required for growth, affects crop growth, and further leads to reduction of yield and quality.

[0003] At present, there is no effective prevention and control measure for replanting disorder. Although chemical control can take effect quickly, long-term and large-scale use of chemical pesticides will cause environmental pollution, and is easy to produce drug resistance and reduce the prevention and control effect. In addition, some existing anti-replanting products mainly supplement chemical elements, especially medium and trace elements, or apply biological organic fertilizer, anti-replanting microbial agent, etc., and have single function, slow effect, short prevention and control time and poor prevention and control effect. These products are mainly applied as base application, and are inconvenient to apply during the growth period of crops, and the addition of chemical elements is easy to cause environmental and soil problems, and may aggravate the occurrence of diseases. In addition, the strains in biological organic fertilizer are mostly directly applied to planting soil, which not only causes competition between beneficial strains and dominant pathogenic bacteria in the soil and produces inhibition effect, but also is difficult to effectively colonize and play a role in the crop plant roots. SUMMARY

[0004] Technical problems to be solved: In view of the above technical problems, the purpose of the present application is to disclose a preparation method and application of an anti-cropping microbial fertilizer, aiming to solve the problems of intensified crop diseases and pests and decreased yield and quality caused by heavy cropping. The preparation steps are as follows: Bacillus subtilis and Bacillus licheniformis are activated and uniformly mixed, and then alpha-lactalbumin solution and calcium chloride solution are added to prepare Bacillus spore aggregates; after polytrehalose solution is added to bentonite, it is ultrasonically treated with humic acid solution to prepare modified polytrehalose-bentonite compound; B. catenulatum and T. harzianum are uniformly mixed with Bacillus spore aggregates, and then wall material solution is added to prepare composite microbial microcapsule preparation; finally, the microcapsules are mixed with corn straw biochar, chitosan and the like to obtain the finished product. The present application improves the stability and viable count of the strains by calcium ion induction and embedding modification with humic acid, activates soil organic matter, nitrogen, phosphorus and potassium nutrients, inhibits pathogenic bacteria, improves the rhizosphere environment of crops, and effectively improves the growth index and quality of red kidney beans and potatoes, and is suitable for heavy-cropping-resistant planting of crops.

[0005] Technical scheme: A preparation method of an anti-cropping microbial fertilizer, comprising the following steps: S1. After Bacillus subtilis and Bacillus licheniformis are activated, they are uniformly mixed at a volume ratio of 1: (2-4) to prepare mixed Bacillus spore liquid, and then the mixed Bacillus spore liquid and alpha-lactalbumin solution with a concentration of 10-30 mg / mL are uniformly mixed at a volume ratio of 1: (1-3), and then calcium chloride solution with a concentration of 1.7-5 mM is added, and stirred at 25-37℃ and 100-200 rpm for 30-90 min, and then centrifuged and washed to obtain Bacillus spore aggregates; S2. Add bentonite to polytrehalose solution with a mass fraction of 2-5wt%, and stir at 180-220 rpm for 30-60 min to prepare polytrehalose-bentonite suspension; mix the polytrehalose-bentonite suspension and humic acid solution with a mass fraction of 1-2wt% at a volume ratio of 1: (5-10), and treat under ultrasonic power of 300-500 W for 15-20 min, and dry to obtain modified polytrehalose-bentonite compound; S3. After B. catenulatum and T. harzianum are activated, they are uniformly mixed with Bacillus spore aggregates to prepare composite bacteria mixed suspension; then the composite bacteria mixed suspension and modified polytrehalose-bentonite compound with a concentration of 10-15wt% are mixed at a volume ratio of (1-3): 1, and stirred at 25-30℃ and 120-200 rpm for 20-40 min, and freeze-dried to obtain composite bacteria microcapsule preparation; S4. Mix 10-20 parts of composite bacteria microcapsule preparation, 20-30 parts of corn straw biochar, 2-4 parts of chitosan, 4.5-6.5 parts of potassium humate, and 1-2 parts of trace element chelate preparation to prepare anti-cropping microbial fertilizer.

[0006] Preferably, the concentration of the Bacillus subtilis liquid in S1 is (0.5-2) x 109 CFU / mL; the concentration of bacillus licheniformis bacterial solution is (1-3.5) x 10 9 CFU / mL; the concentration of bacillus licheniformis bacterial solution is (1-3) x 10 9 CFU / mL.

[0007] Preferably, the amount of calcium chloride solution added in S1 is 1 / 10-1 / 5 of the total volume of bacillus licheniformis bacterial solution and alpha-lactalbumin solution.

[0008] Preferably, the amount of bentonite added in S2 is 1-3 wt% based on the polyfucan solution.

[0009] Preferably, the concentration of bacillus licheniformis bacterial solution in S3 is (1-3) x 10 8 CFU / mL, the concentration of trichoderma harzianum bacterial solution is (2.5-4) x 10 8 CFU / mL.

[0010] Preferably, the ratio of bacillus licheniformis bacterial solution, trichoderma harzianum bacterial solution and bacillus licheniformis aggregate solution in S3 is (0.5-2) mL:(1-3) mL:0.1 g.

[0011] The anti-cropping microbial fertilizer prepared by any of the above methods.

[0012] Preferably, the anti-cropping microbial fertilizer is used in crop planting. Beneficial effects

[0013] 1. The present application uses calcium ions to induce the adsorption of lactalbumin nanotubes on the surface of bacillus subtilis and bacillus licheniformis, forming bacillus licheniformis aggregates, avoiding the influence of different bacterial characteristics. Calcium ions and lactalbumin molecular chain negative groups specifically electrostatically combine, making the protein molecular chain change from the folded state to the extended state, exposing more hydrophobic regions and active sites, and also playing a "bridging role" to connect dispersed proteins into linear polymer chains, and the linear polymer chains curl into closed nanotube structures. The amino and carboxyl groups on the surface of lactalbumin nanotubes and the amino and phosphotransferase phosphate groups in the peptidoglycan of bacillus licheniformis cell wall electrostatically interact and preliminarily combine, and then hydrogen bond, hydrophobic interaction and van der Waals force are used to strengthen the combination, so that a single bacillus licheniformis is combined with multiple lactoferrin nanotubes, and through the "bacteria-nanotube-bacteria" bridging effect, bacillus licheniformis aggregates are formed, ensuring the good stability of the two kinds of bacillus licheniformis.

[0014] 2.The application utilizes humic acid to modify the polyfuscaprene-bentonite gel network to synchronously embed the Aureobasidium pullulans, Trichoderma harzianum and Bacillus polymyxa polymer, and to prepare a composite microbial microcapsule preparation, thereby avoiding the influence of dominant pathogenic bacteria in the heavy-crop soil on the survival of the bacterial species. The polyfuscaprene polysaccharide molecular chain is inserted into the layered structure of the bentonite, the hydroxyl groups thereof form hydrogen bonds with the bentonite sheet layer hydroxyl groups, the molecules self-associate to form crosslinking points, and a porous three-dimensional gel network is formed, thereby providing a binding site for bacterial species loading. The humic acid carboxyl groups form electrostatic salt bonds with the exchangeable cations between the bentonite layers, the carboxyl groups and phenolic hydroxyl groups thereof form hydrogen bonds with the gel network hydroxyl groups, and are combined onto the gel network, thereby improving the water absorption and retention capacity, increasing the active adsorption sites, and enhancing the physical strength, and creating conditions for isolating the interference of pathogenic bacteria. The gel network carboxyl groups are electrostatically attracted to the surface amino groups of the bacterial species, the hydroxyl groups form electrostatic interactions with the surface phosphate groups and carboxyl groups of the bacterial species, and rapid synchronous anchoring is achieved; hydrogen bonds are also formed between the network and the bacterial species, and hydrophobic interactions occur in the hydrophobic regions to strengthen the combination, which is conducive to the formation of a stable embedding structure and reduces the threat of pathogenic bacteria to the survival of the bacterial species.

[0015] 3.The composite microcapsule preparation prepared by the application is used in the heavy-crop soil, on the one hand, the polymer formed by the Bacillus subtilis and the Bacillus licheniformis occupies the ecological niche of the red kidney bean root system and the surrounding soil by virtue of the structural advantage, competes for carbon, nitrogen nutrients and oxygen, reduces the nutrient intake of pathogenic bacteria, and at the same time, the two produce antibacterial substances to destroy the cell walls of pathogenic bacteria; the Trichoderma harzianum acts on the pathogenic bacteria in a parasitic manner, releases hydrolytic enzymes to degrade pathogenic toxins, and the Aureobasidium pullulans metabolically produces antioxidant active substances to reduce the infection channels of pathogenic bacteria, thereby comprehensively improving the stress resistance of the crop root system. On the other hand, the Bacillus subtilis secretes phosphatase to decompose the insoluble organic phosphorus in the planting soil for the crop to absorb, the Bacillus licheniformis synthesizes transport proteins to promote the release of available potassium from the insoluble potassium ore, and the Aureobasidium pullulans activates trace elements to be transported to the root system; the humic acid of the gel network chelates soil nutrients to avoid loss, the polyfuscaprene decomposes to promote the proliferation of the composite bacterial species, and the bentonite improves the soil aggregate structure, which is beneficial to the efficient absorption of nutrients by the crops, the growth of the root system, and the improvement of the yield and quality of the crops. DETAILED DESCRIPTION

[0016] The application will be further described below in conjunction with examples, and the following examples are used to explain the application, but the application is not limited to the following examples: The bacterial species used in the application: Bacillus subtilis CICC22109, Bacillus licheniformis CICC22069, Aureobasidium pullulans CICC2652 and Trichoderma harzianum CICC13010 are all purchased from the China Industrial Microbial Culture Collection Center.

[0017] The activation process of the bacterial species described in the application is as follows: Example 1

[0018] A preparation method of an anti-continuous cropping microbial fertilizer, comprising the following steps: S1. 20 mL of Bacillus subtilis bacterial liquid (bacterial concentration of 1 x 10 9 CFU / mL) and 60 mL of Bacillus licheniformis bacterial liquid (bacterial concentration of 2 x 10 9 CFU / mL) are mixed to prepare a mixed Bacillus bacterial liquid (bacterial concentration of 1.75 x 10 9 CFU / mL), then 30 mL of the mixed Bacillus bacterial liquid and 60 mL of an α-lactalbumin solution with a concentration of 20 mg / mL are mixed and slowly added with 18 mL of a 2 mM calcium chloride solution, stirred at 30°C and 150 rpm for 60 min, centrifuged at 4000 rpm for 8 min and washed with sterile normal saline for 3 times, and dried to obtain Bacillus aggregatum; ​​​​S2. To 100 mL of 3.5% polyfucose solution, 2 g of bentonite was added and stirred at 200 rpm for 30 min to prepare a polyfucose-bentonite suspension; 10 mL of the polyfucose-bentonite suspension and 70 mL of 1.5% humic acid solution were mixed and treated under ultrasonic power of 300 W for 15 min, and dried to obtain a modified polyfucose-bentonite composite; S3. 30 mL of Clonostachys rosea liquid (bacterial concentration of 2 x 10 8 CFU / mL), 30 mL of Trichoderma harzianum (bacterial concentration of 3 x 10 8 CFU / mL) and 1 g of Bacillus sp. aggregate were mixed to prepare a composite bacteria mixed suspension; 40 mL of the composite bacteria mixed suspension and 20 mL of 12 wt% modified polyfucose-bentonite composite solution were mixed and stirred at 30°C and 150 rpm for 30 min, and freeze-dried to obtain a composite bacteria microcapsule preparation; S4. First, 30 g of the composite bacteria microcapsule preparation, 40 g of corn straw biochar 20-30 parts, 6 g of chitosan oligosaccharide, 10 g of mineral source fulvic acid potassium, and 3 g of trace element chelate preparation were weighed; then the composite bacteria microcapsule preparation and corn straw biochar were placed in a horizontal screw belt mixer and stirred and mixed at low speed for 15 min, and then the chitosan oligosaccharide, mineral source fulvic acid potassium and trace element chelate preparation were mixed and uniformly mixed, and packaged to obtain an anti-repeated cropping microbial fertilizer. Example 2

[0019] A method for preparing an anti-repeated cropping microbial fertilizer, comprising the following steps: S1. 20 mL of Bacillus subtilis liquid (bacterial concentration of 2 x 10 9 CFU / mL) and 60 mL of Bacillus licheniformis liquid (bacterial concentration of 1 x 10 9 CFU / mL) were mixed to prepare a mixed Bacillus sp. liquid (bacterial concentration of 1.25 x 10 9 CFU / mL), and then 30 mL of the mixed Bacillus sp. liquid and 60 mL of 20 mg / mL α-lactalbumin solution were mixed and slowly added with 18 mL of 4 mM calcium chloride solution, stirred at 35°C and 150 rpm for 60 min, centrifuged at 3500 rpm for 10 min and washed with sterile physiological saline for 3 times, and then dried to obtain Bacillus sp. aggregate; S2. To 100 mL of 3.5% polyfucose solution, 2 g of bentonite was added and stirred at 200 rpm for 30 min to prepare a polyfucose-bentonite suspension; 10 mL of the polyfucose-bentonite suspension and 70 mL of 1.5% humic acid solution were mixed and treated under ultrasonic power of 300 W for 15 min, and dried to obtain a modified polyfucose-bentonite composite; S3. 30 mL of Gliocladium liquid (bacterial concentration of 2 x 10 8 CFU / mL), 30 mL of Trichoderma harzianum (bacterial concentration of 3 x 10 8 CFU / mL) and 1 g of Bacillus sp. aggregate were uniformly mixed to prepare a composite bacteria mixed suspension; 40 mL of the composite bacteria mixed suspension and 20 mL of a modified polyfusarinid-bentonite compound solution with a concentration of 12 wt% were mixed and stirred at 30°C and 150 rpm for 30 min, and then freeze-dried to obtain a composite bacteria microcapsule preparation; S4. 30 g of the composite bacteria microcapsule preparation, 40 g of corn straw biochar 20-30 parts, 6 g of chitosan oligosaccharide, 10 g of potassium abscisic acid, and 3 g of trace element chelate preparation were weighed; then the composite bacteria microcapsule preparation and the corn straw biochar were placed in a horizontal screw ribbon mixer and stirred and mixed at low speed for 15 min, and then the chitosan oligosaccharide, the potassium abscisic acid, and the trace element chelate preparation were uniformly mixed, and packaged to obtain an anti-cropping microbial fertilizer. Example 3

[0020] A preparation method of an anti-cropping microbial fertilizer, comprising the following steps: S1. 20 mL of Bacillus subtilis liquid (bacterial concentration of 1.5 x 10 9 CFU / mL) and 40 mL of Bacillus licheniformis liquid (bacterial concentration of 2.5 x 10 9 CFU / mL) were uniformly mixed to prepare a mixed Bacillus liquid (bacterial concentration of 2.17 x 10 9 CFU / mL), and then 30 mL of the mixed Bacillus liquid and 60 mL of an α-lactalbumin solution with a concentration of 20 mg / mL were uniformly mixed, and then 13 mL of a 2 mM calcium chloride solution was slowly added, and stirred at 30°C and 150 rpm for 60 min, and then centrifuged at 4000 rpm for 8 min and washed with sterile physiological saline for 3 times, and then dried to obtain Bacillus aggregate; S2. 2 g of bentonite was added to 100 mL of a polyfusarinid solution with a mass fraction of 3.5%, and stirred at 200 rpm for 30 min to prepare a polyfusarinid-bentonite suspension; 10 mL of the polyfusarinid-bentonite suspension and 70 mL of a humic acid solution with a mass fraction of 1.5% were mixed and treated at an ultrasonic power of 300 W for 15 min, and then dried to obtain a modified polyfusarinid-bentonite compound; S3. 30 mL of Gliocladium liquid (bacterial concentration of 2 x 10 8 CFU / mL), 30 mL of Trichoderma harzianum (bacterial concentration of 3 x 10 8The CFU / mL) was mixed with 1g of Bacillus aggregate to prepare a compound bacterial suspension; then 40mL of the compound bacterial suspension and 20mL of a 12wt% modified polytrehalose-bentonite complex solution were mixed and stirred at 30℃ and 150rpm for 30min, and then freeze-dried to obtain the compound bacterial microcapsule formulation. S4. First, weigh out 30g of compound microbial microcapsule preparation, 40g of corn straw biochar (20-30 parts), 6g of chitosan oligosaccharide, 10g of mineral-derived potassium humate, and 3g of trace element chelating preparation. Then, put the compound microbial microcapsule preparation and corn straw biochar into a horizontal ribbon mixer and stir at low speed for 15 minutes. Next, add chitosan oligosaccharide, mineral-derived potassium humate, and trace element chelating preparation and mix well. Package the mixture to make a microbial fertilizer resistant to continuous cropping. Example 4

[0021] A method for preparing a microbial fertilizer resistant to continuous cropping includes the following steps: S1. Add 20 mL of Bacillus subtilis bacterial suspension (bacterial concentration of 1×10⁻⁶) 9 CFU / mL) and 60 mL of Bacillus licheniformis bacterial solution (bacterial concentration of 2×10⁻⁶ CFU / mL) 9 Mix well to prepare a mixed Bacillus bacterial suspension (CFU / mL) with a bacterial concentration of 1.75 × 10⁻⁶ CFU / mL. 9 (CFU / mL), then mix 30 mL of mixed Bacillus bacterial culture and 90 mL of α-lactalbumin solution with a concentration of 10 mg / mL, and slowly add 12 mL of 2 mM calcium chloride solution. Stir at 30 °C and 150 rpm for 60 min, centrifuge at 3500 rpm for 10 min, wash 3 times with sterile physiological saline, and dry to obtain Bacillus aggregates. S2. Add 2g of bentonite to 100mL of a 3.5% (w / w) trehalose solution and stir at 200rpm for 30min to prepare a trehalose-bentonite suspension; mix 10mL of the trehalose-bentonite suspension with 70mL of a 1.5% (w / w) humic acid solution and treat with an ultrasonic power of 400W for 15min, then dry to obtain the modified trehalose-bentonite composite. S3. Add 30 mL of *Agrostis spp.* solution (strain concentration 2 × 10⁻⁶) to the solution. 8 CFU / mL), 30 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL), and 30 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL). 8 The CFU / mL) was mixed with 1g of Bacillus aggregate to prepare a compound bacterial suspension; then 40mL of the compound bacterial suspension and 20mL of a 12wt% modified polytrehalose-bentonite complex solution were mixed and stirred at 30℃ and 150rpm for 30min, and then freeze-dried to obtain the compound bacterial microcapsule formulation. S4. First, 30 g of the composite bacteria microcapsule preparation, 40 g of corn straw biochar 20-30 parts, 6 g of chitosan oligosaccharide, 10 g of mineral fulvic acid potassium, and 3 g of trace element chelate preparation are weighed; then the composite bacteria microcapsule preparation and corn straw biochar are placed in a horizontal screw ribbon mixer and stirred at low speed for 15 min, and then chitosan oligosaccharide, mineral fulvic acid potassium, and trace element chelate preparation are added and mixed evenly to package into a heavy stubble-resistant microbial fertilizer. Example 5

[0022] A method for preparing a heavy stubble-resistant microbial fertilizer, comprising the following steps: S1. 20 mL of Bacillus subtilis bacterial solution (bacterial concentration of 1.5 x 10 9 CFU / mL) and 40 mL of Bacillus licheniformis bacterial solution (bacterial concentration of 2.5 x 10 9 CFU / mL) are mixed to prepare a mixed Bacillus bacterial solution (bacterial concentration of 2.17 x 10 9 CFU / mL), and then 30 mL of the mixed Bacillus bacterial solution and 60 mL of a 20 mg / mL a-lactalbumin solution are mixed and slowly added with 13 mL of a 2 mM calcium chloride solution, stirred at 30°C and 150 rpm for 60 min, centrifuged at 4000 rpm for 8 min, washed with sterile physiological saline for 3 times, and dried to obtain Bacillus aggregates; S2. 2 g of bentonite is added to 100 mL of a 2% polytrehalose solution, stirred at 200 rpm for 30 min to prepare a polytrehalose-bentonite suspension; 10 mL of the polytrehalose-bentonite suspension and 70 mL of a 2% humic acid solution are mixed and treated at an ultrasonic power of 400 W for 15 min, and dried to obtain a modified polytrehalose-bentonite composite; S3. 30 mL of Microdochium denticulatum bacterial solution (bacterial concentration of 2 x 10 8 CFU / mL), 30 mL of Trichoderma harzianum (bacterial concentration of 3 x 10 8 CFU / mL), and 1 g of Bacillus aggregates are mixed to prepare a composite bacteria mixed suspension; 40 mL of the composite bacteria mixed suspension and 20 mL of a 12 wt% modified polytrehalose-bentonite composite solution are mixed and stirred at 30°C and 150 rpm for 30 min, and freeze-dried to obtain a composite bacteria microcapsule preparation; S4. First, 30 g of the composite bacteria microcapsule preparation, 40 g of corn straw biochar 20-30 parts, 6 g of chitosan oligosaccharide, 10 g of mineral fulvic acid potassium, and 3 g of trace element chelate preparation are weighed; then the composite bacteria microcapsule preparation and corn straw biochar are placed in a horizontal screw ribbon mixer and stirred at low speed for 15 min, and then chitosan oligosaccharide, mineral fulvic acid potassium, and trace element chelate preparation are added and mixed evenly to package into a heavy stubble-resistant microbial fertilizer. Example 6

[0023] A method for preparing a microbial fertilizer resistant to continuous cropping includes the following steps: S1. Add 20 mL of Bacillus subtilis bacterial suspension (bacterial concentration of 1.5 × 10⁻⁶). 9 CFU / mL) and 40 mL of Bacillus licheniformis bacterial suspension (strain concentration of 2.5 × 10⁻⁶ CFU / mL) 9 Mix well to prepare a mixed Bacillus bacterial suspension (CFU / mL) with the bacterial concentration of 2.17 × 10⁻⁶ CFU / mL. 9 (CFU / mL), then mix 30 mL of mixed Bacillus bacterial culture and 60 mL of α-lactalbumin solution with a concentration of 20 mg / mL, and slowly add 13 mL of 2 mM calcium chloride solution. Stir at 30 °C and 150 rpm for 60 min, centrifuge at 4000 rpm for 8 min, wash 3 times with sterile physiological saline, and dry to obtain Bacillus aggregates. S2. Add 2g of bentonite to 100mL of 5% polytrehalose solution and stir at 200rpm for 30min to prepare a polytrehalose-bentonite suspension; mix 10mL of polytrehalose-bentonite suspension with 50mL of 1.5% humic acid solution and treat with ultrasonic power of 300W for 15min, then dry to obtain the modified polytrehalose-bentonite composite. S3. Add 30 mL of *Agrostis spp.* solution (strain concentration 2 × 10⁻⁶) to the solution. 8 CFU / mL), 30 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL), and 30 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL). 8 The CFU / mL) was mixed with 1g of Bacillus aggregate to prepare a compound bacterial suspension; then 40mL of the compound bacterial suspension and 20mL of a 12wt% modified polytrehalose-bentonite complex solution were mixed and stirred at 30℃ and 150rpm for 30min, and then freeze-dried to obtain the compound bacterial microcapsule formulation. S4. First, weigh out 30g of compound microbial microcapsule preparation, 40g of corn straw biochar (20-30 parts), 6g of chitosan oligosaccharide, 10g of mineral-derived potassium humate, and 3g of trace element chelating preparation. Then, put the compound microbial microcapsule preparation and corn straw biochar into a horizontal ribbon mixer and stir at low speed for 15 minutes. Next, add chitosan oligosaccharide, mineral-derived potassium humate, and trace element chelating preparation and mix well. Package the mixture to make a microbial fertilizer resistant to continuous cropping. Example 7

[0024] A method for preparing a microbial fertilizer resistant to continuous cropping includes the following steps: S1. Add 20 mL of Bacillus subtilis bacterial suspension (bacterial concentration of 1.5 × 10⁻⁶). 9 CFU / mL) and 40 mL of Bacillus licheniformis bacterial suspension (strain concentration of 2.5 × 10⁻⁶ CFU / mL) 9Mix well to prepare a mixed Bacillus bacterial suspension (CFU / mL) with the bacterial concentration of 2.17 × 10⁻⁶ CFU / mL. 9 (CFU / mL), then mix 30 mL of mixed Bacillus bacterial culture and 60 mL of α-lactalbumin solution with a concentration of 20 mg / mL, and slowly add 13 mL of 2 mM calcium chloride solution. Stir at 35℃ and 150 rpm for 60 min, centrifuge at 3500 rpm for 10 min, wash 3 times with sterile physiological saline, and dry to obtain Bacillus aggregates. S2. Add 2g of bentonite to 100mL of a 3.5% (w / w) trehalose solution and stir at 200rpm for 30min to prepare a trehalose-bentonite suspension; mix 10mL of the trehalose-bentonite suspension with 70mL of a 1.5% (w / w) humic acid solution and treat with an ultrasonic power of 400W for 15min, then dry to obtain the modified trehalose-bentonite composite. S3. Add 30 mL of *Agropyron cristatum* solution (inoculum concentration of 1.5 × 10⁻⁶). 8 CFU / mL), 30 mL of Trichoderma harzianum (inoculum concentration of 4×10⁻⁶ CFU / mL), 8 The CFU / mL) was mixed with 1g of Bacillus aggregate to prepare a compound bacterial suspension; then 40mL of the compound bacterial suspension and 20mL of a 12wt% modified polytrehalose-bentonite complex solution were mixed and stirred at 30℃ and 150rpm for 30min, and then freeze-dried to obtain the compound bacterial microcapsule formulation. S4. First, weigh out 30g of compound microbial microcapsule preparation, 40g of corn straw biochar (20-30 parts), 6g of chitosan oligosaccharide, 10g of mineral-derived potassium humate, and 3g of trace element chelating preparation. Then, put the compound microbial microcapsule preparation and corn straw biochar into a horizontal ribbon mixer and stir at low speed for 15 minutes. Next, add chitosan oligosaccharide, mineral-derived potassium humate, and trace element chelating preparation and mix well. Package the mixture to make a microbial fertilizer resistant to continuous cropping. Example 8

[0025] A method for preparing a microbial fertilizer resistant to continuous cropping includes the following steps: S1. Add 20 mL of Bacillus subtilis bacterial suspension (bacterial concentration of 1.5 × 10⁻⁶). 9 CFU / mL) and 40 mL of Bacillus licheniformis bacterial suspension (strain concentration of 2.5 × 10⁻⁶ CFU / mL) 9 Mix well to prepare a mixed Bacillus bacterial suspension (CFU / mL) with the bacterial concentration of 2.17 × 10⁻⁶ CFU / mL. 9(CFU / mL), then mix 30 mL of mixed Bacillus bacterial culture and 60 mL of α-lactalbumin solution with a concentration of 20 mg / mL, and slowly add 13 mL of 2 mM calcium chloride solution. Stir at 30 °C and 150 rpm for 60 min, centrifuge at 4000 rpm for 8 min, wash 3 times with sterile physiological saline, and dry to obtain Bacillus aggregates. S2. Add 2g of bentonite to 100mL of a 3.5% (w / w) trehalose solution and stir at 200rpm for 30min to prepare a trehalose-bentonite suspension; mix 10mL of the trehalose-bentonite suspension with 70mL of a 1.5% (w / w) humic acid solution and treat with an ultrasonic power of 300W for 15min, then dry to obtain the modified trehalose-bentonite composite. S3. Add 20 mL of *Agrostis spp.* solution (strain concentration 2 × 10⁻⁶) to the solution. 8 CFU / mL), 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL), and 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL). 8 The CFU / mL) was mixed with 1g of Bacillus aggregate to prepare a compound bacterial suspension; then 40mL of the compound bacterial suspension and 20mL of a 12wt% modified polytrehalose-bentonite complex solution were mixed and stirred at 30℃ and 150rpm for 30min, and then freeze-dried to obtain the compound bacterial microcapsule formulation. S4. First, weigh out 30g of compound microbial microcapsule preparation, 40g of corn straw biochar (20-30 parts), 6g of chitosan oligosaccharide, 10g of mineral-derived potassium humate, and 3g of trace element chelating preparation. Then, put the compound microbial microcapsule preparation and corn straw biochar into a horizontal ribbon mixer and stir at low speed for 15 minutes. Next, add chitosan oligosaccharide, mineral-derived potassium humate, and trace element chelating preparation and mix well. Package the mixture to make a microbial fertilizer resistant to continuous cropping. Example 9

[0026] A method for preparing a microbial fertilizer resistant to continuous cropping includes the following steps: S1. Add 20 mL of Bacillus subtilis bacterial suspension (bacterial concentration of 1.5 × 10⁻⁶). 9 CFU / mL) and 40 mL of Bacillus licheniformis bacterial suspension (strain concentration of 2.5 × 10⁻⁶ CFU / mL) 9 Mix well to prepare a mixed Bacillus bacterial suspension (CFU / mL) with the bacterial concentration of 2.17 × 10⁻⁶ CFU / mL. 9 (CFU / mL), then mix 30 mL of mixed Bacillus bacterial culture and 60 mL of α-lactalbumin solution with a concentration of 20 mg / mL, and slowly add 13 mL of 2 mM calcium chloride solution. Stir at 30 °C and 150 rpm for 60 min, centrifuge at 4000 rpm for 8 min, wash 3 times with sterile physiological saline, and dry to obtain Bacillus aggregates. S2. Add 2g of bentonite to 100mL of a 3.5% (w / w) trehalose solution and stir at 200rpm for 30min to prepare a trehalose-bentonite suspension; mix 10mL of the trehalose-bentonite suspension with 70mL of a 1.5% (w / w) humic acid solution and treat with an ultrasonic power of 300W for 15min, then dry to obtain the modified trehalose-bentonite composite. S3. Add 20 mL of *Agrostis spp.* solution (strain concentration 2 × 10⁻⁶) to the solution. 8 CFU / mL), 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL), and 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL). 8 The CFU / mL) was mixed with 1g of Bacillus aggregate to prepare a compound bacterial suspension; then 40mL of the compound bacterial suspension and 20mL of a 12wt% modified polytrehalose-bentonite complex solution were mixed and stirred at 30℃ and 150rpm for 30min, and then freeze-dried to obtain the compound bacterial microcapsule formulation. S4. First, weigh 40g of compound microbial microcapsule preparation, 50g of corn straw biochar (20-30 parts), 6g of chitosan oligosaccharide, 10g of mineral-derived potassium humate, and 3g of trace element chelating preparation; then put the compound microbial microcapsule preparation and corn straw biochar into a horizontal ribbon mixer and stir at low speed for 15 minutes. Then add chitosan oligosaccharide, mineral-derived potassium humate, and trace element chelating preparation and mix well. Package the mixture to make a microbial fertilizer resistant to continuous cropping.

[0027] Comparative Example 1

[0028] The difference between this comparative example and Example 8 is that α-lactalbumin is replaced with chitosan.

[0029] A method for preparing a microbial fertilizer resistant to continuous cropping includes the following steps: S1. Add 20 mL of Bacillus subtilis bacterial suspension (bacterial concentration of 1.5 × 10⁻⁶). 9 CFU / mL) and 40 mL of Bacillus licheniformis bacterial suspension (strain concentration of 2.5 × 10⁻⁶ CFU / mL) 9 Mix well to prepare a mixed Bacillus bacterial suspension (CFU / mL) with the bacterial concentration of 2.17 × 10⁻⁶ CFU / mL. 9 (CFU / mL), then mix 30 mL of mixed Bacillus bacterial culture and 60 mL of chitosan solution with a concentration of 20 mg / mL, and slowly add 13 mL of 2 mM calcium chloride solution. Stir at 30 °C and 150 rpm for 60 min, centrifuge at 4000 rpm for 8 min, wash 3 times with sterile physiological saline, and dry to obtain Bacillus aggregates. S2. Add 2g of bentonite to 100mL of a 3.5% (w / w) trehalose solution and stir at 200rpm for 30min to prepare a trehalose-bentonite suspension; mix 10mL of the trehalose-bentonite suspension with 70mL of a 1.5% (w / w) humic acid solution and treat with an ultrasonic power of 300W for 15min, then dry to obtain the modified trehalose-bentonite composite. S3. Add 20 mL of *Agrostis spp.* solution (strain concentration 2 × 10⁻⁶) to the solution. 8 CFU / mL), 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL), and 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL). 8 The CFU / mL) was mixed with 1g of Bacillus aggregate to prepare a compound bacterial suspension; then 40mL of the compound bacterial suspension and 20mL of a 12wt% modified polytrehalose-bentonite complex solution were mixed and stirred at 30℃ and 150rpm for 30min, and then freeze-dried to obtain the compound bacterial microcapsule formulation. S4. First, weigh out 30g of compound microbial microcapsule preparation, 40g of corn straw biochar (20-30 parts), 6g of chitosan oligosaccharide, 10g of mineral-derived potassium humate, and 3g of trace element chelating preparation. Then, put the compound microbial microcapsule preparation and corn straw biochar into a horizontal ribbon mixer and stir at low speed for 15 minutes. Next, add chitosan oligosaccharide, mineral-derived potassium humate, and trace element chelating preparation and mix well. Package the mixture to make a microbial fertilizer resistant to continuous cropping. Comparative Example 2

[0030] The difference between this comparative example and Example 8 is that α-lactalbumin and calcium chloride are not added.

[0031] A method for preparing a microbial fertilizer resistant to continuous cropping includes the following steps: S1. Add 20 mL of Bacillus subtilis bacterial suspension (bacterial concentration of 1.5 × 10⁻⁶). 9 CFU / mL) and 40 mL of Bacillus licheniformis bacterial suspension (strain concentration of 2.5 × 10⁻⁶ CFU / mL) 9 Mix well to prepare a mixed Bacillus bacterial suspension (CFU / mL) with the bacterial concentration of 2.17 × 10⁻⁶ CFU / mL. 9 (CFU / mL) S2. Add 2g of bentonite to 100mL of a 3.5% (w / w) trehalose solution and stir at 200rpm for 30min to prepare a trehalose-bentonite suspension; mix 10mL of the trehalose-bentonite suspension with 70mL of a 1.5% (w / w) humic acid solution and treat with an ultrasonic power of 300W for 15min, then dry to obtain the modified trehalose-bentonite composite. S3. Add 20 mL of *Agrostis spp.* solution (strain concentration 2 × 10⁻⁶) to the solution. 8 CFU / mL), 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL), and 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL). 8The CFU / mL solution was mixed with 20 mL of mixed Bacillus bacterial solution to prepare a compound bacterial mixture; then 40 mL of the compound bacterial mixture and 20 mL of 12 wt% modified polytrehalose-bentonite complex solution were mixed and stirred at 30 °C and 150 rpm for 30 min, and then freeze-dried to obtain the compound bacterial microcapsule formulation. S4. First, weigh out 30g of compound microbial microcapsule preparation, 40g of corn straw biochar (20-30 parts), 6g of chitosan oligosaccharide, 10g of mineral-derived potassium humate, and 3g of trace element chelating preparation. Then, put the compound microbial microcapsule preparation and corn straw biochar into a horizontal ribbon mixer and stir at low speed for 15 minutes. Next, add chitosan oligosaccharide, mineral-derived potassium humate, and trace element chelating preparation and mix well. Package the mixture to make a microbial fertilizer resistant to continuous cropping. Comparative Example 3

[0032] The difference between this comparative example and Example 8 is that humic acid modification was not used.

[0033] A method for preparing a microbial fertilizer resistant to continuous cropping includes the following steps: S1. Add 20 mL of Bacillus subtilis bacterial suspension (bacterial concentration of 1.5 × 10⁻⁶). 9 CFU / mL) and 40 mL of Bacillus licheniformis bacterial suspension (strain concentration of 2.5 × 10⁻⁶ CFU / mL) 9 Mix well to prepare a mixed Bacillus bacterial suspension (CFU / mL) with the bacterial concentration of 2.17 × 10⁻⁶ CFU / mL. 9 (CFU / mL), then mix 30 mL of mixed Bacillus bacterial culture and 60 mL of α-lactalbumin solution with a concentration of 20 mg / mL, and slowly add 13 mL of 2 mM calcium chloride solution. Stir at 30 °C and 150 rpm for 60 min, centrifuge at 4000 rpm for 8 min, wash 3 times with sterile physiological saline, and dry to obtain Bacillus aggregates. S2. Add 2g of bentonite to 100mL of a 3.5% (w / w) trehalose solution and stir at 200rpm for 30min to prepare a trehalose-bentonite suspension; S3. Add 20 mL of *Agrostis spp.* solution (strain concentration 2 × 10⁻⁶) to the solution. 8 CFU / mL), 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL), and 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL). 8 A compound bacterial suspension was prepared by mixing 1 g of Bacillus aggregate with 40 mL of the compound bacterial suspension and 20 mL of polytrehalose-bentonite suspension at 30 °C and 150 rpm for 30 min, and then freeze-dried to obtain the compound bacterial microcapsule formulation. S4. First, weigh out 30g of compound microbial microcapsule preparation, 40g of corn straw biochar (20-30 parts), 6g of chitosan oligosaccharide, 10g of mineral-derived potassium humate, and 3g of trace element chelating preparation. Then, put the compound microbial microcapsule preparation and corn straw biochar into a horizontal ribbon mixer and stir at low speed for 15 minutes. Next, add chitosan oligosaccharide, mineral-derived potassium humate, and trace element chelating preparation and mix well. Package the mixture to make a microbial fertilizer resistant to continuous cropping. Comparative Example 4

[0034] The difference between this comparative example and Example 8 is that only the compound bacterial mixture was used to prepare the anti-replanting microbial fertilizer.

[0035] A method for preparing a microbial fertilizer resistant to continuous cropping includes the following steps: S1. Add 20 mL of Bacillus subtilis bacterial suspension (bacterial concentration of 1.5 × 10⁻⁶). 9 CFU / mL) and 40 mL of Bacillus licheniformis bacterial suspension (strain concentration of 2.5 × 10⁻⁶ CFU / mL) 9 Mix well to prepare a mixed Bacillus bacterial suspension (CFU / mL) with the bacterial concentration of 2.17 × 10⁻⁶ CFU / mL. 9 (CFU / mL) S2. Add 2g of bentonite to 100mL of a 3.5% (w / w) trehalose solution and stir at 200rpm for 30min to prepare a trehalose-bentonite suspension; mix 10mL of the trehalose-bentonite suspension with 70mL of a 1.5% (w / w) humic acid solution and treat with an ultrasonic power of 300W for 15min, then dry to obtain the modified trehalose-bentonite composite. S3. Add 20 mL of *Agrostis spp.* solution (strain concentration 2 × 10⁻⁶) to the solution. 8 CFU / mL), 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL), and 20 mL of Trichoderma harzianum (inoculum concentration of 3×10⁻⁶ CFU / mL). 8 A compound bacterial mixture was prepared by mixing (CFU / mL) with 20 mL of mixed Bacillus bacterial solution. S4. First, take 30mL of compound bacterial mixture, 20-30 portions of 40g corn straw biochar, 6g chitosan oligosaccharide, 10g mineral-derived potassium humate, and 3g trace element chelating agent; then put the compound bacterial microcapsule preparation and corn straw biochar into a horizontal ribbon mixer and stir at low speed for 15 minutes. Then add chitosan oligosaccharide, mineral-derived potassium humate and trace element chelating agent and mix well. Package to make anti-replanting microbial fertilizer.

[0036] Performance testing Effective viable bacteria count, moisture content, pH and fineness According to the "Inspection Procedures for Microbial Fertilizer Products" (NY / T2321-2013), the effective viable bacteria count of microbial fertilizers is determined using the plate count method.

[0037] Moisture content According to the "Inspection Procedures for Microbial Fertilizer Products" (NY / T2321-2013), the moisture content of microbial fertilizers is determined by the direct drying method.

[0038] pH According to the "NY / T2321-2013 Inspection Procedure for Microbial Fertilizer Products", the pH of microbial fertilizers is determined using a pH meter.

[0039] Fineness According to the "Inspection Procedures for Microbial Fertilizer Products" (NY / T2321-2013), the fineness of microbial fertilizers is determined by the dry sieving method.

[0040] Table 1. Effective viable count, moisture content, pH, and fineness of Examples 1-8 and Comparative Examples 1-4

[0041] As shown in Table 1, the effective viable count, moisture content, pH, and fineness of Examples 1-9 were all superior to those of Comparative Examples 1-4. This indicates that the use of calcium chloride to induce α-lactalbumin to form nanotubes through calcium ion bridging, which then form electrostatic bonds and hydrogen bonds with the Bacillus cell wall, reducing interference from differences in characteristics between strains. The humic acid-modified trehalose-bentonite encapsulation to construct a porous three-dimensional gel network isolates the strains from external conditions, ensuring good survival stability and thus guaranteeing the quality stability of the microbial fertilizer. In contrast, Comparative Examples 1-4, due to the substitution of raw materials, omission of key steps, or simplification of processes, compromised strain protection and product uniformity, resulting in microbial fertilizers of lower quality than those of Examples 1-9. Example 10

[0042] Experiment on the Application Effect of Anti-Continuous Cropping Microbial Fertilizer in Red Kidney Bean Cultivation A red kidney bean planting experiment was conducted using a scheme that "for every 100 kg of red kidney bean seeds produced, the required nitrogen (N) is approximately 5.49 kg, the required phosphorus (P₂O₅) is approximately 1.33 kg, and the required potassium (K₂O) is approximately 4.90 kg". 30 mu (approximately 2 hectares) of soil from continuous red kidney bean planting was randomly divided into six treatment groups: a blank control group, Example 9 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, and Comparative Example 4 group. Each treatment covered an area of ​​0.33 hectares. 2 (5 mu), the planting density of red kidney beans was 185,000 plants / hm. 2 The row spacing is 50cm, the plant spacing is 30cm, and the fertilization treatment plan is shown in Table 2.

[0043] Table 2 Fertilization treatment schemes for each group in the red kidney bean planting experiment.

[0044] Note: Phosphorus nitrate (N≥26.5%, P2O5≥11.5%), urea (N≥46%).

[0045] After the red kidney bean planting experiment was completed, the soil physicochemical indicators, red kidney bean growth indicators, and quality indicators were analyzed to evaluate the effect of the anti-replanting microbial fertilizer.

[0046] (1) Soil nutrient determination Soil samples from the top 0-20cm layer were taken before and after the experiment using the serpentine marking method. The samples were thoroughly mixed, air-dried, and then used for testing. The contents of organic matter, available nitrogen, available phosphorus, and available potassium in the soil were then determined.

[0047] Table 3. Contents of organic matter, available nitrogen, available phosphorus, and available potassium in the soil after application of microbial fertilizer. Table 3 shows that the soil nutrient content of the anti-replanting microbial fertilizer prepared in Example 9 was higher than that of the blank control group and comparative groups 1-4. This indicates that the microbial fertilizer can significantly increase the content of organic matter, available nitrogen, available phosphorus, and available potassium in the soil, effectively improving the nutrient status of soils subjected to continuous cropping. Compared with other treatment groups, the Example 9 group performed best in all indicators, indicating that the synergistic effect of functional bacteria and organic carriers in its formula is more conducive to nutrient activation and retention, creating a favorable soil environment for the growth of red kidney beans.

[0048] (2) Growth indicators During the maturity period of red kidney beans, 20 uniform and representative red kidney bean plants were selected from each treatment, and their plant height and pod length were measured with a measuring tape.

[0049] Protein and fat For each treatment, 200g of red kidney bean sample was weighed to determine its protein and fat content. Protein was determined using the fully automated Kjeldahl nitrogen determination method, and fat was determined using the Soxhlet extraction method.

[0050] Table 4. Plant height, pod length, protein and fat content of red kidney beans.

[0051] Table 4 shows that the red kidney beans in Example 9 group were significantly higher than those in other groups in terms of plant height, pod length, protein content, and fat content. Specifically, plant height increased by 24.4% compared to the control group, pod length increased by 24.2%, protein content increased by 14.0%, and fat content increased by 20.6%. This indicates that applying this microbial fertilizer not only improves soil nutrients but also effectively promotes the growth and development of red kidney bean plants and enhances the nutritional quality of the seeds. While the comparative groups showed some improvement, their effects were not as significant as those in Example 9 group, further validating the superiority and stability of this method. Example 11

[0052] Experiment on the Application Effect of Anti-Continuous Cropping Microbial Fertilizer in Potato Planting Thirty mu (approximately 6.7 hectares) of soil from continuous cropping was randomly divided into six treatment groups: a blank control group, Example 9 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, and Comparative Example 4 group. The experimental area for each treatment was 0.33 hectares. 2 (5 mu), row spacing 0.6m, plant spacing 0.3m, planting density is 60,000 plants / hm. 2 From the date of planting, compound fertilizer (N:P2O5:K2O=11:17:22) and anti-continuous cropping microbial fertilizer were applied to the experimental field once before sowing according to the experimental design dosage (Table 5). Topdressing was applied to the five treatment groups in a timely manner during the potato tuber enlargement period. All other cultivation, irrigation and field management measures were carried out uniformly and consistently according to conventional methods.

[0053] Table 5. Fertilization treatment schemes for each group in the potato planting experiment.

[0054] After the potato planting trial, the growth and quality indicators of the potatoes were analyzed to evaluate the effect of the anti-replanting microbial fertilizer.

[0055] (1) Growth indicators Potato plant height and emergence rate were measured, with a sampling area of ​​0.5m × 2m. Plant height was measured using a ruler, from the base of the plant stem to the growing point. Emergence rate was calculated based on the number of seedlings in the sampling area.

[0056] (2) Quality Indicators The dry matter content of potatoes was determined by the dry weight method; the protein content was determined by the Kjeldahl nitrogen determination method; the starch content was determined by the anthrone method; the vitamin C content was determined by the 2,6-dichlorophenolindophenol titration method; and the reducing sugar content was determined by the 3,5-dinitrosalicylic acid colorimetric method (DNS).

[0057] Table 6. Growth and quality indicators of potatoes

[0058] Table 6 shows that the plant height, emergence rate, and quality indicators of Group 9 were superior to other groups. The plant height reached 98.65 cm, and the emergence rate was 92.56%, significantly higher than the blank control group and other comparative groups. The protein content was 2.85 g / 100 g, the vitamin C content was 45.83 mg / 100 g, and the reducing sugar content was the lowest at 0.21 g / 100 g, indicating that the anti-replanting microbial fertilizer effectively promoted potato growth and improved its nutritional quality. Meanwhile, the dry matter content of Group 9 was 20.85 g / 100 g, slightly higher than the control group and other comparative groups, indicating that the fertilizer also had a positive effect on improving the accumulation of nutrients in potato tubers. The indicators of each comparative group were close to those of the blank control group, indicating that a single component or the absence of key microbial species significantly weakened the fertilizer effect. In summary, the anti-replanting microbial fertilizer effectively improved potato growth vigor and nutritional quality by optimizing the rhizosphere microecology, and has significant application value.

[0059] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention, or modify them into equivalent embodiments, without departing from the spirit and technical essence of the present invention. Therefore, any simple modifications, equivalent substitutions, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the content of the technical solutions of the present invention, shall still fall within the scope of protection of the present invention.

Claims

1. A method for preparing an anti-crop-rotation microbial fertilizer, characterized by, Comprising the following steps: S1. After activation, mix Bacillus subtilis and Bacillus licheniformis in a volume ratio of 1: (1.5-4) to prepare a mixed Bacillus bacteria solution, then mix the mixed Bacillus bacteria solution and an α-lactalbumin solution with a concentration of 10-30 mg / mL in a volume ratio of 1: (1-3), and then add a calcium chloride solution with a concentration of 1.7-5 mM, stir at 25-37°C, 100-200 rpm for 30-90 min, and centrifuge and wash to obtain a Bacillus aggregate; S2. Add 1-3.5 wt% bentonite to a polytrehalose solution with a mass fraction of 2-5 wt%, and stir at 180-220 rpm for 30-60 min to prepare a polytrehalose-bentonite suspension; mix the polytrehalose-bentonite suspension and a humic acid solution with a mass fraction of 1-2 wt% in a volume ratio of 1: (5-10), and treat under ultrasonic power of 300-500 W for 15-20 min, and dry to obtain a modified polytrehalose-bentonite composite; S3. After activation, mix Bacillus subtilis and Bacillus licheniformis with the Bacillus aggregate to prepare a composite bacteria mixed suspension; then mix the composite bacteria mixed suspension and a modified polytrehalose-bentonite composite solution with a concentration of 10-15 wt% in a volume ratio of (1-3): 1, stir at 25-30°C, 120-200 rpm for 20-40 min, and freeze-dry to obtain a composite bacteria microcapsule preparation; S4. Mix 10-20 parts of the composite bacteria microcapsule preparation, 20-30 parts of corn straw biochar, 2-4 parts of chitosan oligosaccharide, 4.5-6.5 parts of potassium abscisic acid, and 1-2 parts of trace element chelate preparation to prepare an anti-cropping microbial fertilizer.

2. The method of claim 1, wherein the method is characterized by: The concentration of the Bacillus subtilis bacterial solution in S1 is (0.5-3) x 10 9 CFU / mL; the concentration of the Bacillus licheniformis bacterial solution is (1-3.5) x 10 9 CFU / mL; and the concentration of the Bacillus mixtus bacterial solution is (1-3) x 10 9 CFU / mL.

3. The method of claim 1, wherein the method is characterized by: The amount of calcium chloride solution added in S1 is 1 / 10-1 / 5 of the total volume of the mixed Bacillus bacteria solution and the α-lactalbumin solution.

4. The method of claim 1, wherein the method is characterized by: The concentration of the S3 G. tenuissima bacteria solution is (1-3) x 10 8 The concentration of the Trichoderma harzianum bacteria solution is (2.5-4) x 10 8 CFU / mL.

5. The method of claim 1, wherein the method is characterized by: The ratio of the Bacillus subtilis solution, the Bacillus licheniformis solution, and the Bacillus aggregate solution in S3 is (0.5-2) mL: (1-3) mL: 0.1 g.

6. The anti-cropping microbial fertilizer prepared by the method of any one of claims 1-5.

7. The use of the anti-cropping microbial fertilizer of claim 6 in crop planting.

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