Multifunctional coating rod medicine fertilizer nutrient solution and preparation method and application thereof
By combining microbial agents such as Bacillus lateralis, Burkholderia ulmoides, and Trichoderma pleurotus with ingredients such as earthworm protein peptides, a multifunctional stalk-applied fertilizer solution has been developed, solving the problems of single-effect pesticide application and slow tree recovery in apple trees. This solution achieves a comprehensive effect of disease control, tree recovery, and increased fruit yield.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SHANDONG ZHONGNONG YISHENGYUAN BIOTECHNOLOGY CO LTD
- Filing Date
- 2026-04-07
- Publication Date
- 2026-06-26
AI Technical Summary
Existing apple tree stalk coating agents have limited effectiveness in controlling canker disease, resulting in slow tree recovery. Furthermore, long-term use of chemical agents leads to drug resistance and microecological imbalance, making it difficult to simultaneously promote disease control, tree recovery, and increased fruit yield.
A microbial agent composed of Bacillus lateralis, Burkholderia ulmoides, and Trichoderma pleurotus is combined with earthworm protein peptides, Laminaria japonica extract, and other ingredients to form a multifunctional stalk-applied fertilizer solution. Through the synergistic effect of the three, it promotes tree healing, enhances disease resistance, and improves nutrient absorption.
It effectively prevents apple tree rot, promotes tree healing and fruit enlargement, increases yield and quality, and is environmentally friendly, without affecting the environment or fruit safety.
Smart Images
Figure CN122277334A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of bio-organic fertilizer technology, specifically relating to a multifunctional stem-coated medicated fertilizer nutrient solution, its preparation method, and its application. Background Technology
[0002] Apple tree canker, caused by fungi of the genus *Ceratophyllum*, is one of the most serious trunk and branch diseases affecting the apple industry. It often leads to bark rot, weakened tree vigor, and even the death of the entire tree, causing significant economic losses to fruit growers. Currently, control of this disease mainly relies on chemical pesticides such as tebuconazole and thiophanate-methyl applied to the trunk or sprayed. However, long-term use of chemical agents alone not only easily leads to drug resistance in pathogens, causing a gradual decline in control effectiveness, but also disrupts the microecological balance of the tree surface and orchard soil, posing potential risks to the environment and fruit safety. Furthermore, traditional trunk coating products often have limited functionality, focusing only on sterilization while neglecting post-disease recovery and nutrient replenishment. They are unable to simultaneously control the disease and promote callus formation and rapid tree rejuvenation, leading to frequent disease recurrence and limited impact on fruit yield and quality improvement.
[0003] In recent years, with the promotion of green agriculture concepts, the use of beneficial microorganisms and their metabolites in combination with nutrients for disease control has become a research hotspot. However, existing microbial agents or fertilizer products often suffer from weak colonization ability and low nutrient synergistic utilization rates. Specifically, single strains are unable to simultaneously exert multiple effects such as highly efficient antagonism against pathogens, induction of disease resistance, and promotion of growth; simple mixing of conventional fertilizers and microbial agents easily leads to a decrease in microbial activity; and the lack of active substances that can be quickly absorbed by the bark results in slow tree recovery after application. Therefore, developing a multifunctional bark coating product that integrates efficient disease prevention, improved nutrient absorption, and enhanced quality and yield is of great significance for achieving the green and sustainable development of the apple industry. Summary of the Invention
[0004] The purpose of this invention is to provide a multifunctional stalk-coated fertilizer solution, its preparation method, and its application, which effectively treats apple tree rot and improves fruit quality and yield.
[0005] To achieve the above-mentioned technical objectives, the technical solution adopted by the present invention is as follows: A multifunctional nutrient solution for coating stems with pesticides and fertilizers comprises the following raw materials in parts by weight: 5-10 parts wood vinegar, 8-15 parts microbial inoculant, 8-12 parts earthworm protein peptide, 10-20 parts *Alternaria alternifolia* extract, 10-18 parts potassium humate, 5-8 parts lecithin, 3-6 parts sodium carboxymethyl cellulose, 5-12 parts chitosan, 10-15 parts urea, 5-10 parts potassium dihydrogen phosphate, 3-8 parts chelated trace element mixture, and 30-50 parts water.
[0006] Furthermore, the microbial inoculant includes *Bacillus laterosporus*, *U. uram*, and *Trichoderma pleurotus*. *Bacillus laterosporus* was purchased from the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.15087 and original accession date of February 15, 2015; *U. uram* was purchased from the CGMCC with accession number CGMCC No. 1.10201 and original accession date of August 18, 2009; and *Trichoderma pleurotus* was purchased from the CGMCC with accession number CGMCC No. 3.12978 and original accession date of March 26, 2009.
[0007] Furthermore, the preparation method of the microbial inoculant is as follows: (1) Prepare a bacterial suspension from the lyophilized powder of Bacillus laterosporus. Take 100 μL of the above bacterial suspension and add it to nutrient gravy agar medium and incubate for 48 h. Pick a single colony and inoculate it into nutrient gravy liquid medium. Incubate at 30 °C and 160 rpm until OD reaches 0.5. 600 =0.6 to obtain seed culture, inoculate the seed culture into a small seed tank at an inoculum rate of 5%, and incubate at 30℃ and 160rpm until OD. 600 =3.0, yielding the fermentation broth of Bacillus laterosporus; (2) Prepare a bacterial suspension from the lyophilized powder of Burkholderia ulmoides. Take 100 μL of the above bacterial suspension and add it dropwise to tryptone-soybean agar medium and incubate for 48 h. Pick a single colony and inoculate it into tryptone-soybean liquid medium. Incubate at 30 °C and 180 rpm until OD reaches 0.5. 600 =0.6 to obtain seed culture, inoculate the seed culture into a small seed tank at an inoculum rate of 5%, and incubate at 30℃ and 180rpm until OD. 600 =3.0, yielding Ulam's Burkholderia para. (3) After activating Trichoderma pleurotus, inoculate it into PDA medium and culture at 25°C until the spores mature. Wash the spores with sterile water, disperse them, and prepare a spore suspension. Place the suspension in a small fermenter at an inoculation rate of 10% and resuspend the spores in sterile water to achieve a spore concentration of 1×10⁻⁶. 8 A suspension of *Trichoderma pleurotus* spores was obtained by measuring 1 spore per mL. (4) Mix the fermentation broth of Bacillus lateralis, the fermentation broth of Burkholderia ulmoides and the spore suspension of Trichoderma pleuropneumoniae in a volume ratio of 1:1:1.
[0008] Furthermore, the preparation method of the chelated trace element mixture is as follows: Add 100 parts of deionized water to the reaction vessel and heat to 65°C. Add 80 parts of disodium ethylenediaminetetraacetate and stir until completely dissolved. Adjust the pH of the reaction vessel solution to 6.0 with 10% sulfuric acid solution. Add 22 parts of ferrous sulfate heptahydrate, 12 parts of zinc sulfate monohydrate, 2 parts of copper sulfate pentahydrate, 6 parts of manganese sulfate monohydrate, 6 parts of borax, and 0.3 parts of ammonium molybdate to the reaction vessel in sequence. Stir until completely dissolved after each addition before adding the next. After all components are completely dissolved, maintain 65°C and stir at 200 r / min for 30 minutes. Cool to room temperature and filter to obtain a chelated trace element mixture.
[0009] A method for preparing a multifunctional stem-coated fertilizer nutrient solution includes the following steps: First, prepare the chelated trace element mixture and microbial agent separately. Add water to the preparation tank and heat to 40-50℃. While stirring, add potassium humate, urea, and potassium dihydrogen phosphate in sequence and stir until completely dissolved. Continue stirring and add wood vinegar, earthworm protein peptide, algae extract, lecithin, and chitin. Stir evenly to ensure that each component is fully dissolved and dispersed. Continue stirring and add sodium carboxymethyl cellulose to fully dissolve and disperse it. Then add the chelated trace element mixture and stir evenly. Reduce the system temperature to 30℃, add the prepared microbial agent, and stir evenly. Filter the evenly mixed solution through a 200-mesh sieve to obtain the multifunctional coating plant fertilizer nutrient solution.
[0010] This invention also provides the application of the multifunctional stem-coating fertilizer solution, which is used to prevent and control apple tree diseases and improve fruit quality and yield.
[0011] Furthermore, the disease in question is apple tree rot.
[0012] Trichoderma pleurotus can secrete chitinase and other cell wall degrading enzymes, which directly dissolve the cell walls of the pathogens causing rot, leading to the breakage and death of the pathogen hyphae. After the coating is applied to the trunk, Trichoderma pleurotus can quickly colonize the bark surface and wounds, competing for space and nutrients, forming a biological protective film that prevents pathogen infection. It can also promote the formation of callus tissue in the tree, accelerate wound healing, and reduce recurrence.
[0013] Bacillus laterosporus directly inhibits the growth of apple tree canker pathogens by producing lipopeptide antibacterial substances, reducing branch and trunk infection, inducing systemic disease resistance in fruit trees, enhancing the disease resistance of the bark and the tree itself, and reducing the risk of reinfection. In addition, Bacillus laterosporus has nitrogen-fixing capabilities, which can help improve nitrogen utilization, and secretes auxin-like substances to promote callus formation in roots and branches, thus facilitating lesion healing and tree recovery.
[0014] Burkholderia ulmoides is capable of producing indoleacetic acid. After stem coating, its metabolites can stimulate the overall physiological metabolism of the tree, promoting cell division during the budding and young fruit stages of apple trees, laying the foundation for the number of cells needed for later fruit enlargement. This strain enhances the root absorption capacity and promotes photosynthesis, transforming weak trees into strong ones, achieving a virtuous cycle of nourishing the tree with the fungus and resisting disease with the tree.
[0015] The three strains can regulate the microbial community structure on the surface of branches and trunks, inhibit the growth of harmful bacteria, promote the colonization of beneficial bacteria, form a benign micro-ecological environment, and further enhance the overall effect of the nutrient solution.
[0016] This invention innovatively incorporates earthworm protein peptides, which are rich in small-molecule amino acids, polypeptides, and various active substances. These peptides are easily absorbed by the bark of apple tree branches and can quickly replenish the tree's nutrients, enhance tree growth, and provide nutritional support for callus formation. Earthworm protein peptides can promote the absorption and conversion of nitrogen, phosphorus, potassium, and chelated trace elements in the formula, further improving overall fertilizer efficiency and providing sufficient nutritional support for apple fruit enlargement, quality improvement, and increased yield. Its active ingredients can stimulate the apple tree's own defense system, helping to enhance the tree's resistance to cold, drought, and pathogen infection, further strengthening the tree's stress resistance and reducing the impact of adverse environments and diseases on the tree.
[0017] Beneficial effects This invention provides a multifunctional medicated fertilizer solution that can effectively prevent and control apple tree canker, induce disease resistance, improve nutrient absorption, and achieve integrated disease resistance, tree care, and high yield.
[0018] This invention, through the scientific combination of three specific bacterial strains, works synergistically with the raw materials in the formula to solve the problems of existing apple tree stalk coating agents, such as single disease resistance, low fertilizer utilization rate, and slow tree recovery. While effectively preventing and controlling apple tree rot, it can also significantly promote fruit enlargement, increase yield and quality, improve tree vigor, and is green and environmentally friendly, with no adverse effects on the environment and fruit, and has broad application prospects. Attached Figure Description
[0019] Figure 1 This is a diagram showing the antagonism between the microbial inoculant and the pathogen causing apple tree rot in Example 2 of the present invention. Detailed Implementation
[0020] The technical solution of the present invention will be further described below with reference to specific embodiments, but it is not limited thereto.
[0021] Example 1 A multifunctional nutrient solution for coating stems with pesticides and fertilizers comprises the following raw materials in parts by weight: 5 parts wood vinegar, 8 parts microbial agent, 8 parts earthworm protein peptide, 10 parts *Alternaria alternifolia* extract, 10 parts potassium humate, 5 parts lecithin, 3 parts sodium carboxymethyl cellulose, 5 parts chitin, 10 parts urea, 5 parts potassium dihydrogen phosphate, 3 parts chelated trace element mixture, and 30 parts water.
[0022] The microbial inoculant includes *Bacillus laterosporus*, *Burkholderia ulmoides*, and *Trichoderma pylori*, with the preservation numbers of *Bacillus laterosporus* (CGMCC No. 1.15087), *Burkholderia ulmoides* (CGMCC No. 1.10201), and *Trichoderma pylori* (CGMCC No. 3.12978) being CGMCC No. 3.12978.
[0023] The preparation method of the microbial inoculant is as follows: (1) Prepare a bacterial suspension from the lyophilized powder of Bacillus laterosporus. Take 100 μL of the above bacterial suspension and add it to nutrient gravy agar medium and incubate for 48 h. Pick a single colony and inoculate it into nutrient gravy liquid medium. Incubate at 30 °C and 160 rpm until OD reaches 0.5. 600 =0.6 to obtain seed culture, inoculate the seed culture into a small seed tank at an inoculum rate of 5%, and incubate at 30℃ and 160rpm until OD. 600 =3.0, yielding the fermentation broth of Bacillus laterosporus; (2) Prepare a bacterial suspension from the lyophilized powder of Burkholderia ulmoides. Take 100 μL of the above bacterial suspension and add it dropwise to tryptone-soybean agar medium and incubate for 48 h. Pick a single colony and inoculate it into tryptone-soybean liquid medium. Incubate at 30 °C and 180 rpm until OD reaches 0.5. 600 =0.6 to obtain seed culture, inoculate the seed culture into a small seed tank at an inoculum rate of 5%, and incubate at 30℃ and 180rpm until OD. 600 =3.0, yielding Ulam's Burkholderia para. (3) After activating Trichoderma pleurotus, inoculate it into PDA medium and culture at 25°C until the spores mature. Wash the spores with sterile water, disperse them, and prepare a spore suspension. Place the suspension in a small fermenter at an inoculation rate of 10% and resuspend the spores in sterile water to achieve a spore concentration of 1×10⁻⁶. 8 A suspension of *Trichoderma pleurotus* spores was obtained by measuring 1 spore per mL. (4) Mix the fermentation broth of Bacillus lateralis, the fermentation broth of Burkholderia ulmoides and the spore suspension of Trichoderma pleuropneumoniae in a volume ratio of 1:1:1.
[0024] The preparation method of the chelated trace element mixture is as follows: Add 100 parts of deionized water to the reaction vessel and heat to 65°C. Add 80 parts of disodium ethylenediaminetetraacetate and stir until completely dissolved. Adjust the pH of the reaction vessel solution to 6.0 with 10% sulfuric acid solution. Add 22 parts of ferrous sulfate heptahydrate, 12 parts of zinc sulfate monohydrate, 2 parts of copper sulfate pentahydrate, 6 parts of manganese sulfate monohydrate, 6 parts of borax, and 0.3 parts of ammonium molybdate to the reaction vessel in sequence. Stir until completely dissolved after each addition before adding the next. After all components are completely dissolved, maintain 65°C and stir at 200 r / min for 30 minutes. Cool to room temperature and filter to obtain a chelated trace element mixture.
[0025] A method for preparing a multifunctional stem-coated fertilizer nutrient solution includes the following steps: First, prepare the chelated trace element mixture and microbial agent separately. Add water to the preparation tank and heat to 40-50℃. While stirring, add potassium humate, urea, and potassium dihydrogen phosphate in sequence and stir until completely dissolved. Continue stirring and add wood vinegar, earthworm protein peptide, algae extract, lecithin, and chitin. Stir evenly to ensure that each component is fully dissolved and dispersed. Continue stirring and add sodium carboxymethyl cellulose to fully dissolve and disperse it. Then add the chelated trace element mixture and stir evenly. Reduce the system temperature to 30℃, add the prepared microbial agent, and stir evenly. Filter the evenly mixed solution through a 200-mesh sieve to obtain the multifunctional coating plant fertilizer nutrient solution.
[0026] Example 2 A multifunctional nutrient solution for coating stems with medicinal fertilizer comprises the following raw materials in parts by weight: 7 parts wood vinegar, 12 parts microbial agent, 10 parts earthworm protein peptide, 15 parts *Alternaria latifolia* extract, 14 parts potassium humate, 7 parts lecithin, 4 parts sodium carboxymethyl cellulose, 9 parts chitin, 12 parts urea, 8 parts potassium dihydrogen phosphate, 5 parts chelated trace element mixture, and 40 parts water.
[0027] The microbial inoculant includes *Bacillus laterosporus*, *Burkholderia ulmoides*, and *Trichoderma pylori*, with the preservation numbers of *Bacillus laterosporus* (CGMCC No. 1.15087), *Burkholderia ulmoides* (CGMCC No. 1.10201), and *Trichoderma pylori* (CGMCC No. 3.12978) being CGMCC No. 3.12978.
[0028] The preparation method of the microbial inoculant is as follows: (1) Prepare a bacterial suspension from the lyophilized powder of Bacillus laterosporus. Take 100 μL of the above bacterial suspension and add it to nutrient gravy agar medium and incubate for 48 h. Pick a single colony and inoculate it into nutrient gravy liquid medium. Incubate at 30 °C and 160 rpm until OD reaches 0.5. 600 =0.6 to obtain seed culture, inoculate the seed culture into a small seed tank at an inoculum rate of 5%, and incubate at 30℃ and 160rpm until OD.600 =3.0, yielding the fermentation broth of Bacillus laterosporus; (2) Prepare a bacterial suspension from the lyophilized powder of Burkholderia ulmoides. Take 100 μL of the above bacterial suspension and add it dropwise to tryptone-soybean agar medium and incubate for 48 h. Pick a single colony and inoculate it into tryptone-soybean liquid medium. Incubate at 30 °C and 180 rpm until OD reaches 0.5. 600 =0.6 to obtain seed culture, inoculate the seed culture into a small seed tank at an inoculum rate of 5%, and incubate at 30℃ and 180rpm until OD. 600 =3.0, yielding Ulam's Burkholderia para. (3) After activating Trichoderma pleurotus, inoculate it into PDA medium and culture at 25°C until the spores mature. Wash the spores with sterile water, disperse them, and prepare a spore suspension. Place the suspension in a small fermenter at an inoculation rate of 10% and resuspend the spores in sterile water to achieve a spore concentration of 1×10⁻⁶. 8 A suspension of *Trichoderma pleurotus* spores was obtained by measuring 1 spore per mL. (4) Mix the fermentation broth of Bacillus lateralis, the fermentation broth of Burkholderia ulmoides and the spore suspension of Trichoderma pleuropneumoniae in a volume ratio of 1:1:1.
[0029] The preparation method of the chelated trace element mixture is as follows: Add 100 parts of deionized water to the reaction vessel and heat to 65°C. Add 80 parts of disodium ethylenediaminetetraacetate and stir until completely dissolved. Adjust the pH of the reaction vessel solution to 6.0 with 10% sulfuric acid solution. Add 22 parts of ferrous sulfate heptahydrate, 12 parts of zinc sulfate monohydrate, 2 parts of copper sulfate pentahydrate, 6 parts of manganese sulfate monohydrate, 6 parts of borax, and 0.3 parts of ammonium molybdate to the reaction vessel in sequence. Stir until completely dissolved after each addition before adding the next. After all components are completely dissolved, maintain 65°C and stir at 200 r / min for 30 minutes. Cool to room temperature and filter to obtain a chelated trace element mixture.
[0030] A method for preparing a multifunctional stem-coated fertilizer nutrient solution includes the following steps: First, prepare the chelated trace element mixture and microbial agent separately. Add water to the preparation tank and heat to 40-50℃. While stirring, add potassium humate, urea, and potassium dihydrogen phosphate in sequence and stir until completely dissolved. Continue stirring and add wood vinegar, earthworm protein peptide, algae extract, lecithin, and chitin. Stir evenly to ensure that each component is fully dissolved and dispersed. Continue stirring and add sodium carboxymethyl cellulose to fully dissolve and disperse it. Then add the chelated trace element mixture and stir evenly. Reduce the system temperature to 30℃, add the prepared microbial agent, and stir evenly. Filter the evenly mixed solution through a 200-mesh sieve to obtain the multifunctional coating plant fertilizer nutrient solution.
[0031] Example 3 A multifunctional nutrient solution for coating stems with medicinal fertilizer comprises the following raw materials in parts by weight: 10 parts wood vinegar, 15 parts microbial agent, 12 parts earthworm protein peptide, 20 parts *Alternaria alternifolia* extract, 18 parts potassium humate, 8 parts lecithin, 6 parts sodium carboxymethyl cellulose, 12 parts chitin, 15 parts urea, 10 parts potassium dihydrogen phosphate, 8 parts chelated trace element mixture, and 50 parts water.
[0032] The microbial inoculant includes *Bacillus laterosporus*, *Burkholderia ulmoides*, and *Trichoderma pylori*, with the preservation numbers of *Bacillus laterosporus* (CGMCC No. 1.15087), *Burkholderia ulmoides* (CGMCC No. 1.10201), and *Trichoderma pylori* (CGMCC No. 3.12978) being CGMCC No. 3.12978.
[0033] The preparation method of the microbial inoculant is as follows: (1) Prepare a bacterial suspension from the lyophilized powder of Bacillus laterosporus. Take 100 μL of the above bacterial suspension and add it to nutrient gravy agar medium and incubate for 48 h. Pick a single colony and inoculate it into nutrient gravy liquid medium. Incubate at 30 °C and 160 rpm until OD reaches 0.5. 600 =0.6 to obtain seed culture, inoculate the seed culture into a small seed tank at an inoculum rate of 5%, and incubate at 30℃ and 160rpm until OD. 600 =3.0, yielding the fermentation broth of Bacillus laterosporus; (2) Prepare a bacterial suspension from the lyophilized powder of Burkholderia ulmoides. Take 100 μL of the above bacterial suspension and add it dropwise to tryptone-soybean agar medium and incubate for 48 h. Pick a single colony and inoculate it into tryptone-soybean liquid medium. Incubate at 30 °C and 180 rpm until OD reaches 0.5. 600 =0.6 to obtain seed culture, inoculate the seed culture into a small seed tank at an inoculum rate of 5%, and incubate at 30℃ and 180rpm until OD. 600 =3.0, yielding Ulam's Burkholderia para. (3) After activating Trichoderma pleurotus, inoculate it into PDA medium and culture at 25°C until the spores mature. Wash the spores with sterile water, disperse them, and prepare a spore suspension. Place the suspension in a small fermenter at an inoculation rate of 10% and resuspend the spores in sterile water to achieve a spore concentration of 1×10⁻⁶. 8 A suspension of *Trichoderma pleurotus* spores was obtained by measuring 1 spore per mL. (4) Mix the fermentation broth of Bacillus lateralis, the fermentation broth of Burkholderia ulmoides and the spore suspension of Trichoderma pleuropneumoniae in a volume ratio of 1:1:1.
[0034] The preparation method of the chelated trace element mixture is as follows: Add 100 parts of deionized water to the reaction vessel and heat to 65°C. Add 80 parts of disodium ethylenediaminetetraacetate and stir until completely dissolved. Adjust the pH of the reaction vessel solution to 6.0 with 10% sulfuric acid solution. Add 22 parts of ferrous sulfate heptahydrate, 12 parts of zinc sulfate monohydrate, 2 parts of copper sulfate pentahydrate, 6 parts of manganese sulfate monohydrate, 6 parts of borax, and 0.3 parts of ammonium molybdate to the reaction vessel in sequence. Stir until completely dissolved after each addition before adding the next. After all components are completely dissolved, maintain 65°C and stir at 200 r / min for 30 minutes. Cool to room temperature and filter to obtain a chelated trace element mixture.
[0035] A method for preparing a multifunctional stem-coated fertilizer nutrient solution includes the following steps: First, prepare the chelated trace element mixture and microbial agent separately. Add water to the preparation tank and heat to 40-50℃. While stirring, add potassium humate, urea, and potassium dihydrogen phosphate in sequence and stir until completely dissolved. Continue stirring and add wood vinegar, earthworm protein peptide, algae extract, lecithin, and chitin. Stir evenly to ensure that each component is fully dissolved and dispersed. Continue stirring and add sodium carboxymethyl cellulose to fully dissolve and disperse it. Then add the chelated trace element mixture and stir evenly. Reduce the system temperature to 30℃, add the prepared microbial agent, and stir evenly. Filter the evenly mixed solution through a 200-mesh sieve to obtain the multifunctional coating plant fertilizer nutrient solution.
[0036] Comparative Example 1 Compared with Example 2, this comparative example uses the same raw materials and steps as Example 2, except that the microbial agents used are Bacillus retroflexus fermentation broth and Burkholderia ulmosporum fermentation broth at a volume ratio of 1:1.
[0037] Comparative Example 2 Compared with Example 2, this comparative example uses the same raw materials and steps as Example 2, except that the microbial agent used is Bacillus retroflexus fermentation broth and Trichoderma spore suspension with a volume ratio of 1:1.
[0038] Comparative Example 3 Compared with Example 2, this comparative example uses the same raw materials and steps as Example 2, except that the microbial inoculants used were Burkholderia ulmoides fermentation broth and Trichoderma spore suspension in a volume ratio of 1:1.
[0039] Comparative Example 4 Compared with Example 2, this comparative example is identical to Example 2 except that no earthworm protein peptides are added. All other raw materials and steps are the same as in Example 2.
[0040] Performance testing Example 2: Antagonistic Test of Microbial Agents Against Apple Rot Disease A hole was punched in the center of the PDA medium, and 20 μL of the microbial inoculum prepared in Example 2 was inoculated into it. Mycelial cakes with a diameter of 5 mm were prepared from the activated apple tree rot pathogen using a punch. The mycelial cakes were placed on both sides of the well in the PDA medium and incubated at 25°C for 3 days. The antagonistic experiment results are shown below. Figure 1 .
[0041] Depend on Figure 1 It is known that the microbial agents prepared from Bacillus lateralis, Burkholderia ulmoides, and Trichoderma pleurotus used in this invention can effectively inhibit apple tree rot disease.
[0042] Field efficacy trial of apple tree rot disease In mid-to-late September 2024, 10-year-old Fuji apple trees were treated with a stalk coating in an apple orchard with moderate soil fertility and cultivation management. The experiment consisted of eight treatment groups, with ten trees in each group. The treatments used were nutrient solutions prepared in Examples 1-3, Comparative Examples 1-4, and a blank control. The application method was stalk coating on the main trunk and large branches, performed twice on September 15th and 29th, 2024, with the nutrient solution diluted 10 times. Before stalk coating and in the following spring, the number of diseased scars on the main trunk, large branches, and lateral branches of each tree was recorded, and the control effect was calculated. Control effect = (Number of new diseased scars in the control group - Number of new diseased scars in the treatment group) / Number of new diseased scars in the control group × 100%. The statistical results are shown in Table 1.
[0043] Table 1 Field efficacy As shown in Table 1, the nutrient solution of this invention has a control effect of over 60% on apple tree canker, with a maximum control effect of up to 75%, indicating that the nutrient solution of this invention has a good control effect on apple tree canker. However, in comparative examples 1-3 with altered bacterial agent composition, the control effect was significantly reduced because the synergistic effect between the three bacterial strains was disrupted.
[0044] Planting Trial Experimental materials: The apple variety was "Red Fuji," 4-year-old fruit trees with basically uniform growth, and the row spacing was 3m × 2m. The basic physicochemical properties of the tested soil were: pH value 5.32, organic matter content 19.37g / kg, available nitrogen content 109.15 mg / kg, available phosphorus content 30.08mg / kg, and available potassium content 112.46 mg / kg.
[0045] Experimental setup: Nutrient solutions prepared in Examples 1-3 and Comparative Examples 1-4 were used for stem coating. The nutrient solutions prepared for each treatment group were diluted at a mass ratio of 1:30. The stems were coated four times, at the budding stage, flowering stage, young fruit stage, and fruit enlargement stage. The control (CK) was not coated. There were a total of 8 treatment groups, with 6 plants in each treatment group. Conventional field management was maintained.
[0046] Fruit data statistics and harvesting: Statistics were collected after the fruits ripened in October. Fruits of similar shape and size, free from disease, pests, and damage, were selected for harvesting. Immediately after harvesting, the fruits were placed in foam boxes with ice packs and transported to the laboratory, where they were kept fresh at 0°C. Thirty fruits were randomly harvested from each treatment group to determine the weight and diameter of each fruit. The yield of each plot was calculated and converted to yield per acre. The statistical data are shown in Table 2. The internal quality of the fruits was measured. Fruit firmness and flesh crispness were determined according to "Determination of Fruit Firmness" (NY / T 2009—2011). Soluble solids content was determined according to "Determination of Soluble Solids Content in Fruits and Vegetables - Refractometer Method" (NY / T2637—2014). Vitamin C content was determined according to "National Food Safety Standard - Determination of Ascorbic Acid in Food" (GB5009.86—2016). Titratable acid content was determined according to "Determination of Total Acid in Food" (GB / T 12456—2008). The measurement data are shown in Table 3.
[0047] Table 2. Fruit appearance quality and yield of each treatment group As shown in Table 2, the nutrient solution prepared in this invention, after being applied to the stems, significantly increased fruit weight and fruit expansion compared to the control (CK), thereby increasing apple yield. However, comparative examples 1-3, which changed the composition of the inoculant, all showed significant yield reductions to varying degrees, indicating that the fruit expansion effect decreased in the absence of any of the strains selected in this invention. This demonstrates that the three strains selected in this invention are indispensable for increasing apple tree yield.
[0048] Table 3. Internal quality of fruits in each treatment group As shown in Table 3, the fruit firmness, flesh crispness, soluble solids content, and vitamin C content were all improved after using the nutrient solutions prepared in Examples 1-3 of this invention compared to the control (CK), indicating an improvement in fruit quality. However, the apple quality of Comparative Examples 1-3 (with altered bacterial composition in the nutrient solution) and Comparative Example 4 (without earthworm extract) decreased to varying degrees, indicating that the effectiveness of the nutrient solution is weakened if any of the three bacterial strains or earthworm extract are omitted.
[0049] It should be noted that the above embodiments are merely some preferred embodiments of the present invention, and not all embodiments. Obviously, based on the above embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.
Claims
1. A multifunctional stem-applied fertilizer solution, characterized in that, The ingredients include the following parts by weight: 5-10 parts wood vinegar, 8-15 parts microbial inoculant, 8-12 parts earthworm protein peptide, 10-20 parts *Alternaria alternifolia* extract, 10-18 parts potassium humate, 5-8 parts lecithin, 3-6 parts sodium carboxymethyl cellulose, 5-12 parts chitosan, 10-15 parts urea, 5-10 parts potassium dihydrogen phosphate, 3-8 parts chelated trace element mixture, and 30-50 parts water.
2. The multifunctional stem-coating fertilizer and nutrient solution according to claim 1, characterized in that, The microbial inoculant includes *Bacillus laterosporus*, *Burkholderia ulmoides*, and *Trichoderma pylori*, with the preservation numbers of *Bacillus laterosporus* (CGMCC No. 1.15087), *Burkholderia ulmoides* (CGMCC No. 1.10201), and *Trichoderma pylori* (CGMCC No. 3.12978) being CGMCC No. 3.12978.
3. The multifunctional stem-coating fertilizer solution according to claim 1, characterized in that, The preparation method of the microbial inoculant is as follows: (1) Prepare a bacterial suspension from the lyophilized powder of Bacillus laterosporus. Take 100 μL of the above bacterial suspension and add it to nutrient gravy agar medium and incubate for 48 h. Pick a single colony and inoculate it into nutrient gravy liquid medium. Incubate at 30 °C and 160 rpm until OD reaches 0.
5. 600 =0.6 to obtain seed culture, inoculate the seed culture into a small seed tank at an inoculum rate of 5%, and incubate at 30℃ and 160rpm until OD. 600 =3.0, yielding the fermentation broth of Bacillus laterosporus; (2) Prepare a bacterial suspension from the lyophilized powder of Burkholderia ulmoides. Take 100 μL of the above bacterial suspension and add it dropwise to tryptone-soybean agar medium and incubate for 48 h. Pick a single colony and inoculate it into tryptone-soybean liquid medium. Incubate at 30 °C and 180 rpm until OD reaches 0.
5. 600 =0.6 to obtain seed culture, inoculate the seed culture into a small seed tank at an inoculum rate of 5%, and incubate at 30℃ and 180rpm until OD. 600 =3.0, yielding Ulam's Burkholderia para. (3) After activating Trichoderma pleurotus, inoculate it into PDA medium and culture at 25°C until the spores mature. Wash the spores with sterile water, disperse them, and prepare a spore suspension. Place the suspension in a small fermenter at an inoculation rate of 10% and resuspend the spores in sterile water to achieve a spore concentration of 1×10⁻⁶. 8 A suspension of *Trichoderma pleurotus* spores was obtained by measuring 1 spore per mL. (4) Mix the fermentation broth of Bacillus lateralis, the fermentation broth of Burkholderia ulmoides and the spore suspension of Trichoderma pleuropneumoniae in a volume ratio of 1:1:
1.
4. The multifunctional stem-coating fertilizer solution according to claim 1, characterized in that, The preparation method of the chelated trace element mixture is as follows: Add 100 parts of deionized water to the reaction vessel and heat to 65°C. Add 80 parts of disodium ethylenediaminetetraacetate and stir until completely dissolved. Adjust the pH of the reaction vessel solution to 6.0 with 10% sulfuric acid solution. Add 22 parts of ferrous sulfate heptahydrate, 12 parts of zinc sulfate monohydrate, 2 parts of copper sulfate pentahydrate, 6 parts of manganese sulfate monohydrate, 6 parts of borax, and 0.3 parts of ammonium molybdate to the reaction vessel in sequence. Stir until completely dissolved after each addition before adding the next. After all components are completely dissolved, maintain 65°C and stir at 200 r / min for 30 minutes. Cool to room temperature and filter to obtain a chelated trace element mixture.
5. A method for preparing the multifunctional stem-coating fertilizer nutrient solution according to any one of claims 1-4, characterized in that, Preparation includes the following steps: First, prepare the chelated trace element mixture and microbial agent separately. Add water to the preparation tank and heat to 40-50℃. While stirring, add potassium humate, urea, and potassium dihydrogen phosphate in sequence and stir until completely dissolved. Continue stirring and add wood vinegar, earthworm protein peptide, algae extract, lecithin, and chitin. Stir evenly to ensure that each component is fully dissolved and dispersed. Continue stirring and add sodium carboxymethyl cellulose to fully dissolve and disperse it. Then add the chelated trace element mixture and stir evenly. Reduce the system temperature to 30℃, add the prepared microbial agent, and stir evenly. Filter the evenly mixed solution through a 200-mesh sieve to obtain the multifunctional coating plant fertilizer nutrient solution.
6. An application of the multifunctional stem-coating fertilizer nutrient solution according to claim 1, characterized in that, The multifunctional stem coating nutrient solution is used to prevent and control apple tree diseases, and to improve fruit quality and increase yield.
7. The application according to claim 6, characterized in that, The disease in question is apple tree rot.