Special disease-resistant compound microbial fertilizer for tea trees and preparation method thereof

Abstract

The invention discloses a special disease-resistant compound microbial fertilizer for tea trees and a preparation method, and belongs to the technical field of agricultural fertilizers. The bacterial fertilizer consists of a microbial flora, a macro-element fertilizer, a trace element fertilizer and a carrier, wherein the microbial flora contains bacillus, trichoderma and actinomycetes. The preparation method comprises the steps of strain culture, fermentation, drying, crushing and mixing. Through the synergistic effect of various microorganisms, the bacterial fertilizer forms a protective barrier on tea tree root systems, inhibits growth of pathogenic bacteria, induces tea trees to generate resistance and enhances the disease resistance of the tea trees; the fertilizer is rich in various nutrient elements, meets the growth requirements of tea trees and promotes the growth of the tea trees; the soil structure is improved, the soil fertility is improved, and soil sustainable utilization is achieved; the content of nutritional ingredients such as tea polyphenol and amino acid in the tea can be improved, and the tea quality is improved. A plurality of embodiments verify that the bacterial fertilizer can significantly reduce the incidence rate of tea cake disease and tea anthracnose, increase the germination number and length of new shoots of tea trees, improve the amino acid content of tea leaves and the like, and has a good application effect.

Description

Technical Field

[0001] The invention relates to the technical field of agricultural fertilizers, and in particular to a disease-resistant composite microbial fertilizer special for tea trees and a preparation method thereof. Background Art

[0002] Tea trees are an important economic crop in my country. During the tea production process, disease control plays a vital role in the yield and quality of tea. At present, tea gardens mainly rely on chemical pesticides and fertilizers to ensure the growth of tea trees and control diseases. However, the long-term and large-scale use of chemical pesticides will not only lead to excessive pesticide residues in tea leaves, which is harmful to human health, but also damage the ecological environment of tea gardens, cause imbalances in soil microbial communities, and cause soil compaction and decreased fertility. At the same time, the excessive use of conventional fertilizers cannot fully meet the tea trees' demand for nutrients, which in turn affects the quality and yield of tea. Therefore, the development of a green, environmentally friendly and efficient disease-resistant fertilizer specifically for tea trees has become a key issue that needs to be urgently addressed in the field of tea cultivation. Summary of the Invention

[0003] The present invention aims to provide a disease-resistant composite microbial fertilizer specifically for tea trees and a preparation method thereof. Through the synergistic effect of microorganisms and nutrients, the disease resistance of tea trees can be enhanced, the use of chemical pesticides and fertilizers can be reduced, the soil environment can be improved, and thus the quality and yield of tea leaves can be improved.

[0004] The specific technical solutions of the present invention are as follows:

[0005] A disease-resistant composite microbial fertilizer specially used for tea trees is characterized by being composed of the following materials in parts by weight: 2-5 parts of microbial flora, 40-70 parts of macroelement fertilizer, 0.5-3 parts of trace element fertilizer, and 25-58 parts of a carrier.

[0006] As a further improvement of the present invention, the microbial flora is composed of a mixture of 0.5-3 parts of Bacillus, 0.1-1 parts of Trichoderma, and 0.3-0.6 parts of actinomycetes. The Bacillus includes Bacillus subtilis and Bacillus amyloliquefaciens, with the mass percentage of Bacillus subtilis being 0.2%-1.5% and the mass percentage of Bacillus amyloliquefaciens being 0.2%-1.5%. In a preferred range, the mass percentage of Bacillus subtilis is 0.5%-1%, and the mass percentage of Bacillus amyloliquefaciens is 0.5%-1%.

[0007] Furthermore, in the macroelement fertilizer, the mass percentage of nitrogen is 8%-20%, preferably in the range of 12%-16%; the mass percentage of phosphorus is 5%-15%, preferably in the range of 8%-12%; the mass percentage of potassium is 8%-20%, preferably in the range of 12%-16%.

[0008] In trace element fertilizers, the total mass percentage of trace element fertilizers such as iron, zinc, and manganese is 0.5%-3%, and the preferred range is 1%-2%.

[0009] The carrier is a mixture after fermentation of soybean meal, corn flour, wheat bran, etc., and the mass percentage is 25%-58%, and the preferred range is 35%-47%.

[0010] Preparation method

[0011] Strain culture: Bacillus subtilis, Bacillus amyloliquefaciens, Trichoderma, and Actinomycetes are respectively inoculated into the corresponding liquid media. The culture temperature is controlled at 25°C-35°C, the rotation speed is 150-220 r / min, and the culture time is 18-48 h. Under these conditions, a suitable growth environment is provided for the strains to ensure their activity and reproduction speed, so that they reach a sufficient quantity and activity. The preferred culture temperature is 28°C-32°C, the rotation speed is 180-200 r / min, and the time is 24-36 h. Under these conditions, the strains can reproduce efficiently, and high-quality seed liquid can be obtained, laying a foundation for subsequent fermentation. The highly active and sufficient quantity of strains cultivated are the key to subsequent functions such as disease resistance, growth promotion, and soil improvement. For example, highly active Bacillus subtilis can form a biological protective film more quickly when colonizing the tea tree roots subsequently, effectively blocking the invasion of pathogenic bacteria; highly active Actinomycetes produce more growth hormones subsequently, more powerfully promoting the development of tea tree roots.

[0012] Fermentation: The seed liquid is inoculated into a solid fermentation medium containing raw materials such as soybean meal, corn flour, and wheat bran at a certain ratio (volume ratio of (0.8-1.5):1:(0.8-1.5):1), the moisture content is adjusted to 50%-70%, the fermentation temperature is controlled at 28°C-35°C, and the fermentation time is 4-8 d. Solid fermentation can provide a rich nutrient source and a suitable environment, which is conducive to the large-scale reproduction of microorganisms. The proteins and carbohydrates in the raw materials can meet their growth and metabolic needs. The change in the inoculation ratio will affect the relative quantity and synergy of microorganisms, while moisture, temperature, and time determine the growth and metabolite accumulation degree of microorganisms. The preferred inoculation volume ratio is 1:1:1:1, the moisture is 55%-65%, the temperature is 30°C-32°C, and the time is 5-6 d. At this time, each microorganism can fully cooperate, reproduce in large numbers, and produce rich metabolites. During this process, a large number of reproduced Trichoderma will produce more hydrolases, enhancing the ability to dissolve the cell walls of pathogenic bacteria; the synergistic Bacillus and Actinomycetes can produce more antibiotics and growth hormones, providing a guarantee for enhancing the disease resistance of tea trees and promoting growth when the subsequent bacterial fertilizer is used in tea gardens.

[0013] Drying: The fermented product is dried at a low temperature. The drying temperature is controlled below 35°C - 50°C, so that its moisture content is reduced to below 5% - 15%, preferably below 40°C - 45°C, and the moisture is below 8% - 12%. Low-temperature drying can maximize the retention of microbial activity and avoid damage caused by high temperature. A suitable moisture content is conducive to storage and transportation, preventing microbial inactivation or the mildew and deterioration of the bio-fertilizer. Only by ensuring microbial activity can it be ensured that microorganisms continue to play a role in subsequent use. For example, the surviving Bacillus subtilis continues to decompose organic matter in the tea garden soil and improve the soil nutrient structure; the surviving actinomycetes continuously produce growth hormones to assist the growth of tea trees.

[0014] Crushing and mixing: The dried product is crushed, and then chemical fertilizers such as nitrogen, phosphorus, and potassium and trace elements are added and mixed thoroughly to obtain a special anti-disease compound microbial bio-fertilizer for tea trees. Crushing makes the bio-fertilizer particles uniform, facilitating mixing and application with other fertilizers. Mixing ensures the uniform distribution of various nutrient components and microorganisms. The uniformly distributed microorganisms and nutrient components can comprehensively provide nutrients and disease resistance protection for tea trees after being applied to the tea garden. For example, the uniformly distributed trace elements can be absorbed by tea trees in a timely manner and participate in the physiological metabolism of tea trees; the uniformly distributed Trichoderma can contact pathogenic bacteria more comprehensively and inhibit their growth.

[0015] Mechanism of action

[0016] Disease resistance mechanism: Bacillus subtilis and Bacillus amyloliquefaciens can rapidly colonize the roots of tea trees, forming a biological protective film to block the invasion of pathogenic bacteria. At the same time, they produce antibiotics and enzyme substances such as subtilin and chitinase to inhibit the growth of common tea tree pathogenic bacteria such as Colletotrichum gloeosporioides and Exobasidium vexans. Trichoderma has the ability to parasitize and can directly parasitize on the mycelium of pathogenic bacteria, inhibiting the growth of pathogenic bacteria by secreting hydrolases to dissolve the cell wall of pathogenic bacteria. In addition, Trichoderma can also induce systemic resistance in tea trees, stimulate the self-defense mechanism of tea trees, and enhance the disease resistance of tea trees. In practical applications, the incidence of tea trees in tea gardens using this bio-fertilizer is significantly reduced. For example, the incidence of Exobasidium vexans is reduced by 20 - 25% compared with tea gardens without using the bio-fertilizer, and the incidence of Colletotrichum gloeosporioides is reduced by 15 - 20%, effectively reducing the reduction in tea production and the decline in quality caused by diseases.

[0017] Growth-promoting mechanism: Growth hormones such as auxins and cytokinins produced by actinomycetes can promote the growth and development of tea plant roots, increase the root absorption area and absorption capacity, improve the tea plant's utilization of nutrients and water, and thus enhance the tea plant's growth potential. Nitrogen, phosphorus, and potassium in macronutrient fertilizers, as well as iron, zinc, and manganese in trace element fertilizers, participate in the tea plant's physiological metabolism, meet the nutrient needs of the tea plant at different growth stages, and promote its growth and development. After using this bacterial fertilizer, the growth of new tea shoots increased significantly, with new shoot length increasing by 12% compared to the control group. The number of leaves increased, and the leaf area increased. The overall growth of the tea plant became more robust, laying the foundation for high tea yields.

[0018] Soil Improvement Mechanism: Bacillus subtilis and Bacillus amyloliquefaciens can break down organic matter in the soil, converting it into nutrients that can be absorbed and utilized by tea plants, thereby improving the soil's nutrient structure. The carrier, composed of a fermented mixture of soybean meal, corn flour, and bran, not only provides a living environment for microorganisms and participates in the fermentation process, but also increases the soil's organic matter content, improves soil structure, and enhances soil fertility, creating a favorable soil environment for tea plantations. Long-term use of this microbial fertilizer reduces soil bulk density in tea gardens, enhances air permeability and water retention, increases the number of beneficial microorganisms in the soil, and stabilizes the soil microbial community structure, providing better soil conditions for tea plantations, promoting their sustained and healthy growth, and improving the quality and yield stability of tea leaves. Furthermore, the effective release of nutrients in the soil increases fertilizer utilization by tea plants, reducing fertilizer waste and environmental pollution.

[0019] Tea Quality Improvement Mechanism: Due to the improved growing environment for tea trees, their disease resistance is enhanced, and they grow stronger. This also positively impacts the nutritional composition of tea leaves. The use of this microbial fertilizer has increased tea polyphenols by 9% and amino acids by 10%. This results in a mellower taste, richer aroma, and a brighter tea color. This makes tea more competitive in the market and brings higher economic benefits to tea farmers.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] Enhanced disease resistance: Multiple microorganisms work synergistically to form a dense protective barrier around the tea tree's roots. Antibiotics and enzymes produced by Bacillus subtilis and Bacillus amyloliquefaciens directly inhibit the growth and reproduction of common pathogens such as anthracnose and tea cake disease. Trichoderma, through parasitism, destroys the pathogen's structure while simultaneously inducing systemic resistance in the tea tree itself. Practical application has demonstrated a significant reduction in tea tree disease incidence in tea gardens using this microbial fertilizer. For example, the incidence of tea cake disease decreased by approximately 30%, and that of tea anthracnose by approximately 25%. This effectively reduces tea production losses due to disease and reduces the manpower and material costs of disease prevention and control.

[0022] Balanced Nutrition: Bacterial fertilizer is not only rich in macronutrients like nitrogen, phosphorus, and potassium, but also precisely supplements essential trace elements like iron, zinc, and manganese for tea plant growth. Microbial metabolic activity also converts nutrients in the soil that are difficult for the tea plant to absorb into usable forms. This comprehensive and balanced nutritional supply meets the needs of tea plants at different stages of growth. Tea plants grow stronger, with more new shoots, thicker, richer green leaves, and improved photosynthetic efficiency. Compared to tea plants not treated with bacterial fertilizer, the number of new shoots increases by approximately 20%, laying a solid foundation for high-yield and high-quality tea.

[0023] Soil improvement: The microorganisms in the bacterial fertilizer can decompose organic matter in the soil, increase the content of soil organic matter, and improve soil structure. Bacillus subtilis and other bacteria can promote the formation of soil aggregate structure, improve soil permeability and water retention. Long-term use of this bacterial fertilizer can reduce soil bulk density by about 0.1g / cm 3 The porosity increased by about 10%, and the soil microbial community became richer and more stable. Soil fertility was effectively improved, reducing dependence on chemical fertilizers and achieving sustainable soil utilization.

[0024] Improved tea quality: A favorable growing environment and adequate nutrient supply boost the metabolic activity of tea plants. This significantly increases the content of tea polyphenols, amino acids, and other nutrients. For example, tea polyphenols have increased by approximately 15%, and amino acids by approximately 10%. This results in a mellower, fresher flavor and a richer, longer-lasting aroma. Furthermore, the tea leaves become firmer and more uniform in appearance, with a richer color, making them more competitive in the market and generating higher economic benefits for tea farmers. DETAILED DESCRIPTION

[0025] Example 1

[0026] Bacterial culture: Inoculate 10 mL of each of the corresponding 1000 mL liquid culture medium with Bacillus subtilis, Bacillus amyloliquefaciens, Trichoderma, and Actinomycetes. Use a nutrient-rich culture medium that meets the growth requirements of each bacterial species, such as LB medium for Bacillus subtilis and Bacillus amyloliquefaciens, potato dextrose medium for Trichoderma, and Gouldian medium No. 1 for Actinomycetes. Place the inoculated culture medium in a constant temperature shaker, precisely control the culture temperature at 28°C, and maintain a constant rotation speed of 180 rpm. During the culture process, measure the bacterial concentration every 2 hours using the dilution plate method, and plot a growth curve. When the bacterial concentration of Bacillus subtilis and Bacillus amyloliquefaciens reaches \(1×10^{8}-5×10^{8}

[0027] When the concentration of Trichoderma and Actinomycetes reached 5×10^{7}-2×10^{8} cfu / mL\), the culture was considered complete.

[0028] Fermentation: Inoculate the above-prepared seed liquid into 5000 g of a solid fermentation medium containing 30% soybean meal, 20% corn flour, and 50% wheat bran according to a volume ratio of 1:1:1:1. Before inoculation, perform high-temperature sterilization on the solid fermentation medium, and then inoculate it after cooling to room temperature. Spray sterile water into the solid fermentation medium with a sprayer to adjust the moisture content to 60%, and accurately calculate the amount of added water by the weighing method. Place the inoculated solid fermentation medium in a constant-temperature incubator and ferment at 30°C for 5 days. Turn the medium regularly 3 - 4 times a day, with an interval of 3 - 4 h each time, to ensure oxygen supply and uniform temperature. During the fermentation process, use a pH test paper to detect the pH value of the fermentation product every day and record its changes; at the same time, observe the growth status of microorganisms under a microscope every other day. When the fermentation product has a strong fermentation aroma, the pH value is stable at 6.5 - 7.0, and a large number of microorganisms are observed to multiply under the microscope and the accumulation of metabolites reaches a certain level (such as a large number of spores are produced by Bacillus, Trichoderma mycelium covers the medium, and Actinomycetes produce obvious pigments, etc.), it is determined that the fermentation is completed.

[0029] Drying and processing: Transfer the fermentation product to a low-temperature drying oven and dry it at a low temperature below 40°C, carefully turning it with a small spatula every 2 - 3 hours to accelerate the drying process. Use a high-precision moisture meter to detect the moisture content in real-time, and stop drying when the moisture content drops to 10%. Crush the dried product to 80 - 100 mesh with a pulverizer, and then accurately weigh nitrogen, phosphorus, potassium chemical fertilizers and trace elements according to the proportion of the bio-fertilizer formula, put them into a blender and mix well for 45 minutes, and regularly check the mixing uniformity of the materials in the blender during the mixing process.

[0030] Usage method: Select a sunny and soil-moist day before the spring budding of tea trees. Use soil detection equipment in advance to detect the humidity and pH value of the tea garden soil to ensure that the soil conditions are suitable for fertilization. With the tea tree trunk as the center, use a professional circular groove digging tool to dig a circular groove with a depth of 15 - 20 cm around the tea tree at a distance of 30 - 50 cm from the trunk. Apply 0.5 kg of bio-fertilizer to each tea tree, and evenly sprinkle the bio-fertilizer into the circular groove using a quantitative fertilization device. After fertilization, promptly cover the circular groove with soil, with a soil covering thickness of 5 - 8 cm, use a flat tool to compact the covered soil, and then water thoroughly to make the bio-fertilizer fully contact with the soil. After watering, detect the soil humidity again to ensure that the soil humidity remains within the range suitable for the growth of tea trees.

[0031] Disease resistance and nutritional balance data: The incidence of Exobasidium vexans decreased by 22.3% compared to the tea gardens without using the bacterial fertilizer, and the incidence of Colletotrichum camelliae decreased by 16.7%. The number of new shoots of the tea plants increased by 20.4% compared to the control group, the length of the new shoots increased by 12.6%, the leaf thickness increased by 8.1%, the chlorophyll content in the leaves increased by 11.2%, and the amino acid content in the tea leaves increased by 8.9%, as shown in Table 1.

[0032] Example 2

[0033] Strain cultivation: Bacillus subtilis, Bacillus amyloliquefaciens, Trichoderma, and Actinomycetes were respectively inoculated into 1500 mL of the corresponding liquid medium, and the inoculation amounts were 18 mL, 15 mL, 18 mL, and 15 mL of the bacterial liquid in sequence. Optimize the medium formula according to the characteristics of each strain, and adjust the proportion of nutritional components such as the carbon-nitrogen ratio. The culture temperature was controlled at 30 °C, and the rotation speed was 200 r / min. During the cultivation period, the bacterial liquid concentration was measured every 3 h by the dilution coating plate method, and the growth curve was plotted. When the bacterial liquid concentration of Bacillus subtilis and Bacillus amyloliquefaciens reached \(2×10^{8}-8×10^{8} cfu / mL\), and the bacterial liquid concentration of Trichoderma and Actinomycetes reached \(8×10^{7}-3×10^{8} cfu / mL\), it was determined that the strain cultivation was completed.

[0034] Fermentation: The seed liquid was inoculated into 8000 g of a solid fermentation medium containing 25% soybean meal, 25% corn flour, and 50% wheat bran according to a volume ratio of 1.2:1:1.2:1. Before inoculation, the solid fermentation medium was sterilized by high-pressure steam and subjected to a sterility test. The moisture content was adjusted to 55% using a spray device, and a moisture rapid detector was used to monitor it in real time. Fermentation was carried out in a constant temperature incubator at 32 °C for 6 days. Sterile air was introduced for 15 - 20 minutes every 12 hours, and the ventilation volume was precisely controlled by a gas flow meter. The content of metabolites such as antibiotics and growth hormones in the fermentation product was detected by a high-performance liquid chromatograph every day. When the pH value of the fermentation product was stable at 6.5 - 7.0, and the content of metabolites (such as antibiotics, growth hormones, etc.) reached the expected range through detection means such as high-performance liquid chromatography, it was determined that the fermentation was completed.

[0035] Drying and processing: The fermentation product was dried at a temperature below 45 °C, and it was slowly turned over with a stirrer every 3 - 4 hours to avoid damaging the microbial structure. A near-infrared moisture detector was used to detect the moisture content, and the drying ended when the moisture content dropped to 8%. It was pulverized to 100 - 120 meshes, and fertilizers and trace elements were mixed in according to a precisely calculated formula, and stirred and mixed for 60 minutes. During the stirring process, multiple samples were taken for detection to ensure the mixing uniformity.

[0036] Usage method: During the peak growth season of tea trees, select a windless evening. Conduct a fertility test on the tea garden soil in advance to determine the specific location and dosage of fertilization. At the edge of the tea tree crown projection, use a special small shovel to dig 3 - 5 holes with a depth of 15 - 20 cm, and the hole spacing is 20 - 30 cm. Apply 0.4 kg of microbial fertilizer to each tea tree, and evenly put the microbial fertilizer into the holes using a quantitative spoon. After application, cover the soil, with the soil covering thickness of 8 - 10 cm. Use a small compaction tool to compact the covered soil, then water thoroughly. After watering, use a soil moisture sensor to detect the soil humidity to ensure that the microbial fertilizer can quickly integrate into the soil environment.

[0037] Disease resistance and nutrient balance data: The incidence of tea blister blight is reduced by 23.1% compared to the tea garden without using microbial fertilizer, and the incidence of tea anthracnose is reduced by 17.4%. The number of new shoots germinated by tea trees increases by 21.7% compared to the control group, the new shoot length increases by 13.3%, the leaf thickness increases by 9.5%, the chlorophyll content in the leaves increases by 11.8%, and the amino acid content in the tea leaves increases by 9.2%, as shown in Table 1.

[0038] Example 3

[0039] Strain cultivation: Inoculate Bacillus subtilis, Bacillus amyloliquefaciens, Trichoderma, and Actinomycetes into 2000 mL of the corresponding liquid medium respectively, and the inoculation amounts are 20 mL, 20 mL, 15 mL, and 15 mL of bacterial liquid in sequence. The medium uses a customized formula, and the nutrient components are optimized for each strain. For example, an appropriate amount of organic nitrogen source is added for Bacillus strains to promote their growth. The cultivation temperature is set at 31 °C, and the rotation speed is 190 r / min. During the cultivation process, the bacterial liquid concentration is measured every 2.5 hours by the dilution coating plate method, and a growth curve is plotted. When the bacterial liquid concentrations of Bacillus subtilis and Bacillus amyloliquefaciens reach \(3×10^{8}-6×10^{8} cfu / mL\), and the bacterial liquid concentrations of Trichoderma and Actinomycetes reach \(7×10^{7}-2.5×10^{8} cfu / mL\), and the strain viability is good, the strain cultivation is completed.

[0040] Fermentation: According to the volume ratio of 1:1.1:1:1.1, inoculate the cultured seed liquid into 10000 g of a solid fermentation medium containing 28% soybean meal, 22% corn flour, and 50% wheat bran. Use a sprayer to adjust the moisture content to 62%, and use a humidity sensor to monitor it in real time. Ferment in a constant temperature incubator at 31 °C for 5.5 days, turn the medium 4 times a day, and the interval between each turn is 4 hours. At the same time, detect the contents of antibiotics and growth hormones in the fermentation products by enzyme-linked immunosorbent assay (ELISA) every other day. When the fermentation product has a strong smell, the pH value is stable at 6.6 - 6.9, and the ELISA detection shows that the metabolite reaches the expected amount, the fermentation ends.

[0041] Drying and processing: Place the fermented product in a low-temperature drying oven and dry it below 42°C, gently turning it with a stirring paddle every 3 hours. Stop drying when the moisture content drops to 9%. Crush it to 90 - 110 mesh, add nitrogen, phosphorus, potassium chemical fertilizers and trace elements according to the precise formula, and stir and mix for 50 minutes to ensure uniformity.

[0042] Application method: At the initial stage of the new shoots' germination of tea trees, select cloudy days or early and late periods of sunny days. In the rows of tea trees, at a distance of 40 - 60 cm from the tea tree roots, dig strip-shaped trenches with a depth of 18 - 22 cm. Apply 80 kg of the bacterial fertilizer per mu of tea garden, evenly sprinkle it into the trenches, cover it with 8 - 10 cm of soil, and then water it thoroughly immediately.

[0043] Disease resistance and nutrient balance data: The incidence of tea blister blight is reduced by 23.6% compared to the tea garden without using the bacterial fertilizer, and the incidence of tea anthracnose is reduced by 17.2%. The number of new shoots of tea trees increases by 22.8% compared to the control group, and the leaf thickness increases by 10.3%, as shown in Table 1.

[0044] Example 4

[0045] Strain culture: Inoculate Bacillus subtilis, Bacillus amyloliquefaciens, Trichoderma, and Actinomycetes into 1800 mL of the corresponding liquid medium respectively, with the inoculation amounts being 16 mL, 18 mL, 16 mL, and 14 mL of the bacterial solution. The medium is used after high-temperature sterilization and aseptic detection. Control the culture temperature at 32°C and the rotation speed at 210 r / min. Every 3 hours, use the dilution plating method to measure the concentration of the bacterial solution and draw a growth curve. When the concentration of the bacterial solution of Bacillus subtilis and Bacillus amyloliquefaciens reaches \(2.5×10^{8}-7×10^{8}cfu / mL\), and the concentration of the bacterial solution of Trichoderma and Actinomycetes reaches \(6×10^{7}-3×10^{8}cfu / mL\), the strain culture meets the standards.

[0046] Fermentation: Inoculate the seed liquid into 9000 g of solid fermentation medium containing 26% soybean meal, 24% corn flour, and 50% wheat bran at a volume ratio of 1.3:1:1.3:1. Adjust the moisture content to 58% through a drip irrigation system and continuously monitor it with an electronic moisture meter. Ferment in a constant-temperature incubator at 32.5°C for 5.8 days, and introduce sterile air for 20 - 25 minutes every 10 hours. During fermentation, analyze the components and contents of the metabolites every day using high-performance liquid chromatography - mass spectrometry (HPLC-MS). When the pH value of the fermented product stabilizes at 6.5 - 6.8 and the content of the key metabolites detected by HPLC-MS meets the requirements, end the fermentation.

[0047] Drying and processing: Dry the fermented product below 43°C and turn it over every 3.5 hours. Complete drying when the moisture content drops to 11%, crush it to 100 - 130 mesh, add various fertilizers and trace elements according to precise measurement, and stir and mix for 55 minutes.

[0048] Usage method: Select a rainless early morning one month before the tea tree picking. At the drip line of the tea tree crown, dig 4 - 6 radial grooves with a depth of 16 - 20 cm and a length of 30 - 40 cm around the tea tree. Apply 0.45 kg of bacterial fertilizer to each tea tree, evenly sprinkle it into the grooves, then cover the soil with a thickness of 6 - 9 cm, and then water appropriately.

[0049] Disease resistance and nutrient balance data: The incidence of tea blister blight is reduced by 24.2%, and the incidence of tea anthracnose is reduced by 18.5%. The length of the new shoots of the tea tree is expected to increase by 14.8% compared with the control group, and the chlorophyll content in the leaves is increased by 13.1%, as shown in Table 1.

[0050] Example Five

[0051] Strain culture: Inoculate Bacillus subtilis, Bacillus amyloliquefaciens, Trichoderma, and Actinomycetes into 2200 mL of the corresponding liquid medium respectively, and the inoculation amount is 22 mL of the bacterial solution. Fine-tune the nutrient components of the liquid medium to increase the content of trace elements to meet the growth requirements of the strains. Keep the culture temperature at 30.5 °C and the rotation speed at 185 r / min. During the culture period, measure the bacterial solution concentration every 4 hours by the dilution coating plate method and draw a growth curve. When the bacterial solution concentration of Bacillus subtilis and Bacillus amyloliquefaciens reaches \(4×10^{8}-7×10^{8}cfu / mL\), and the bacterial solution concentration of Trichoderma and Actinomycetes reaches \(8×10^{7}-3×10^{8}

[0052] cfu / mL\), the strain culture is completed.

[0053] Fermentation: Inoculate the seed liquid into 12000 g of a solid fermentation medium containing 27% soybean meal, 23% corn flour, and 50% wheat bran according to a volume ratio of 1:1.2:1.2:1. Adjust the moisture content to 63% with an atomizing nozzle and monitor it with a humidity monitor. Ferment in a constant temperature incubator at 31.5 °C for 6.2 days, turn the culture medium 5 times a day, with an interval of 3 hours each time. Detect the types and contents of antibiotics in the fermentation products by thin-layer chromatography every 1.5 days. When the smell of the fermentation product is normal, the pH value is stable at 6.7 - 7.0, and the thin-layer chromatography results show that the metabolic products such as antibiotics are rich, the fermentation is completed.

[0054] Drying and processing: Dry the fermentation product at a temperature below 41 °C and turn it over every 2.5 hours. Stop drying when the moisture content drops to 10%, crush it to 95 - 120 mesh, strictly add nitrogen, phosphorus, and potassium chemical fertilizers and trace elements according to the formula, and stir and mix for 60 minutes.

[0055] Usage method: During the dormant period of tea trees, select a period with suitable soil moisture. In the tea garden, with the tea tree as the center, evenly dig 5 - 7 holes with a depth of 20 - 25 cm within a range of 50 - 70 cm in radius. Apply 90 kg of microbial fertilizer per mu of tea garden. After evenly applying the microbial fertilizer into the holes, cover the soil with 10 - 12 cm of soil, and appropriately water to keep the soil moist.

[0056] Disease resistance and nutrient balance data: The incidence of tea blister blight is reduced by 24.7%, and the incidence of tea anthracnose is reduced by 18.8%. The absorption area of the tea tree roots is expected to increase by 11.4% compared to the control group, and the amino acid content in the tea leaves is increased by 13.6%, as shown in Table 1.

[0057] Table 1: Summary of disease resistance and nutrient balance data

[0058]

Claims

1. A special anti-disease compound microbial fertilizer for tea trees, characterized in that, It consists of the following parts by weight: 2 - 5 parts of microbial flora, 40 - 70 parts of macronutrient chemical fertilizer, 0.5 - 3 parts of micronutrient fertilizer, and 25 - 58 parts of carrier.

2. The special anti-disease compound microbial fertilizer for tea trees according to claim 1, characterized in that The microbial flora is a mixture composed of 0.5 - 3 parts of bacillus, 0.1 - 1 part of trichoderma, and 0.3 - 0.6 part of actinomycetes.

3. The special anti-disease compound microbial fertilizer for tea trees according to claim 2, characterized in that, The bacillus includes bacillus subtilis and bacillus amyloliquefaciens, where the mass percentage of bacillus subtilis is 0.2% - 1.5%, and the mass percentage of bacillus amyloliquefaciens is 0.2% - 1.5%; preferably, the mass percentage of bacillus subtilis is 0.5% - 1%, and the mass percentage of bacillus amyloliquefaciens is 0.5% - 1%.

4. The special anti-disease compound microbial fertilizer for tea trees according to claim 1, characterized in that, In the macronutrient fertilizer, the mass percentage of nitrogen element is 8% - 20%, and the preferred range is 12% - 16%; the mass percentage of phosphorus element is 5% - 15%, and the preferred range is 8% - 12%; the mass percentage of potassium element is 8% - 20%, and the preferred range is 12% - 16%.

5. The special anti-disease compound microbial fertilizer for tea trees according to claim 1, characterized in that, In the micronutrient fertilizer, the total mass percentage of micronutrient fertilizers such as iron, zinc, and manganese is 0.5% - 3%, and the preferred range is 1% - 2%.

6. The special anti-disease compound microbial fertilizer for tea trees according to claim 1, characterized in that The carrier is a mixture after fermentation of soybean meal, corn flour, wheat bran, etc., with a mass percentage of 25% - 58%, and the preferred range is 35% - 47%.

7. A preparation method of the special anti-disease compound microbial fertilizer for tea trees according to any one of claims 1-6, characterized in that, It includes the following steps: Strain cultivation: Bacillus subtilis, Bacillus amyloliquefaciens, Trichoderma, and Actinomycetes are respectively inoculated into the corresponding liquid medium, controlling the culture temperature at 25°C - 35°C, the rotation speed at 150 - 220 r / min, and the culture time at 18 - 48 h; preferably, the culture temperature is 28°C - 32°C, the rotation speed is 180 - 200 r / min, and the time is 24 - 36 h; Fermentation: The cultured seed liquid is inoculated into the solid fermentation medium containing raw materials such as soybean meal, corn flour, and wheat bran according to the volume ratio of (0.8 - 1.5):1:(0.8 - 1.5):1, adjusting the moisture content to 50% - 70%, and fermenting at 28°C - 35°C for 4 - 8 d; preferably, the inoculation volume ratio is 1:1:1:1, the moisture is 55% - 65%, the temperature is 30°C - 32°C, and the time is 5 - 6 d; Drying: The fermentation product is dried at a low temperature, controlling the drying temperature below 35°C - 50°C to reduce the moisture content to below 5% - 15%; preferably, the drying temperature is below 40°C - 45°C, and the moisture is below 8% - 12%; Crushing and mixing: The dried product is crushed, and then nitrogen, phosphorus, and potassium chemical fertilizers and micronutrients are added and mixed evenly.

8. Application of the special anti - disease compound microbial fertilizer for tea trees described in any one of claims 1 - 6 in enhancing the disease - resistance ability of tea trees, promoting the growth of tea trees, improving the tea garden soil environment, and enhancing the quality of tea leaves.

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