A cultivation method for preventing and controlling Cyclocarya paliurus diseases using microbial fertilizer

Abstract

The invention discloses a cultivation method for preventing and controlling Cyclocarya paliurus diseases by using microbial fertilizer, and belongs to the field of agricultural and forestry disease prevention and control. The cultivation method comprises the step of treating Cyclocarya paliurus by using microbial fertilizer; the microbial fertilizer comprises: 30 parts of Bacillus pumilus powder, 50 parts of Bacillus powder and 30-75 parts of Flammulina velutipes residue biochar; the Bacillus pumilus is Bacillus pumilus CICC 22096, and the Bacillus pumilus is Bacillus pumilus ACCC 64091. The invention compounded Bacillus pumilus CICC 22096 and Bacillus pumilus ACCC 64091, and used Flammulina velutipes residue biochar as a carrier, which can provide superior conditions for strain growth and reproduction and the load of metabolites, promote the rapid formation of dominant bacterial communities, and more concentratedly supply the resistance components in the metabolites to plants, promote the rapid absorption and utilization of plants, and lay a foundation for improving plant disease resistance.

Description

Technical Field

[0001] The invention relates to the field of agricultural and forestry disease prevention and control, and in particular to a cultivation method for preventing and controlling Cyclocarya paliurus diseases by utilizing microbial fertilizer. Background Art

[0002] Cyclocarya paliurus, a deciduous tree of the genus Cyclocarya in the Juglandaceae family, produces ornamental fruits resembling strings of copper coins, making it a valuable landscape tree and a valuable timber species. Its young leaves have medicinal properties, including lowering blood pressure, blood sugar, and blood lipids, enhancing immunity, and providing antioxidant and anti-aging benefits. Its wood is also a high-quality furniture material. Due to its diverse applications, market demand for Cyclocarya paliurus is increasing. Therefore, ensuring a continuous supply of Cyclocarya paliurus requires ensuring consistent quality and consistent annual production.

[0003] However, Cyclocarya paliurus is very susceptible to diseases during its seedling stage, especially damping-off and black leaf blight. Among them, damping-off is common in nurseries, and black leaf blight is common in transplanted seedlings after being transplanted from the nursery. Both diseases directly affect the yield and quality of Cyclocarya paliurus. The current methods for preventing and controlling damping-off and black leaf blight mainly include improved variety breeding and chemical control. Improved variety breeding can fundamentally improve the situation where damping-off is prone to occur, but the breeding cycle is long and the investment cost is high; chemical control generally involves disinfecting the nursery before sowing, and using chemical agents such as methyl thiophanate, thiophanate-methyl, and carbendazim to prevent and control damping-off or black leaf blight. Although chemical control is effective, long-term use causes environmental pollution and resistance in pathogenic microorganisms. Therefore, it is very important to find new methods for preventing and controlling Cyclocarya paliurus.

[0004] Microbial fertilizers are a new type of biological fertilizer product that uses the life activities of active microorganisms to provide crops with the nutrients they need. They are a type of fertilizer used in agricultural production (also known as third-generation fertilizers). Microbial fertilizers have been applied to agricultural disease control with significant results. While there are reports of using single microbial agents to control Cyclocarya paliurus diseases, their efficacy is generally limited to 60%-70%, and they struggle to restore the microecological structure. Not all strains are effective in biocontrol, and different strains exhibit significant synergistic effects with plants. As market demand for both the quantity and quality of Cyclocarya paliurus continues to increase, finding single or combined microbial agents with improved biocontrol effectiveness is crucial to addressing the issue of poor disease control in Cyclocarya paliurus. Summary of the Invention

[0005] The present invention aims to provide a cultivation method for preventing and controlling Cyclocarya paliurus diseases using microbial fertilizer, so as to solve the problems existing in the above-mentioned prior art. After compounding Bacillus pumilus CICC 22096 and Bacillus ACCC 64091, Flammulina velutipes residue biochar is used as a carrier, which can provide excellent conditions for the growth and reproduction of strains and the loading of metabolites, promote the rapid formation of dominant bacterial communities, and more concentratedly supply resistance components in the metabolites to plants, promote their rapid absorption and utilization, and lay the foundation for improving plant disease resistance.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] The present invention provides a cultivation method for preventing and controlling Cyclocarya paliurus diseases by utilizing microbial fertilizer. The method comprises the step of treating Cyclocarya paliurus by utilizing the microbial fertilizer. The microbial fertilizer comprises the following components in parts by weight: 30 parts of Bacillus pumilus powder, 50 parts of Bacillus sp. powder, and 30-75 parts of Flammulina velutipes residue biochar. The Bacillus pumilus is Bacillus pumilus CICC 22096, and the Bacillus sp. is Bacillus sp. ACCC64091.

[0008] Optionally, the method of treating Cyclocarya paliurus with the microbial fertilizer includes: applying the microbial fertilizer to the roots of Cyclocarya paliurus during the nursery period or spraying the microbial fertilizer on the leaves of the transplanted Cyclocarya paliurus seedlings.

[0009] Optionally, 5-7 kg of the microbial fertilizer is used per mu of Cyclocarya paliurus seedlings. This dosage is mainly for Cyclocarya paliurus seedlings in the nursery stage or transplanted from the nursery. If it is for the prevention and treatment of black blight on adult leaves, the dosage can be adjusted appropriately according to needs.

[0010] The present invention also provides a microbial fertilizer comprising the following components in parts by weight: 30 parts of Bacillus pumilus powder, 50 parts of Bacillus subtilis powder and 30-75 parts of Flammulina velutipes residue biochar;

[0011] The Bacillus pumilus is Bacillus pumilus CICC 22096, and the Bacillus is Bacillus ACCC64091.

[0012] The present invention also provides a method for preparing the microbial fertilizer, which is characterized by comprising uniformly mixing the components in the formula.

[0013] Optionally, the Bacillus pumilus powder is obtained by activating Bacillus pumilus CICC 22096, fermenting it in a nutrient broth medium at 35-37° C. for 3 days, collecting the fermentation liquid, and freeze-drying it;

[0014] The bacillus powder is obtained by activating bacillus ACCC 64091, fermenting and culturing it in a nutrient broth medium at 35-37° C. for 3 days, collecting the fermentation liquid, and freeze-drying it.

[0015] Optionally, the preparation step of the Flammulina velutipes residue biochar includes: pyrolyzing the Flammulina velutipes residue at a high temperature of 300-400°C for 30 minutes, and then reducing the high temperature to 100-200°C for 1 hour to obtain mushroom residue-based biochar.

[0016] The present invention also provides application of the microbial fertilizer in preventing and treating Cyclocarya paliurus diseases.

[0017] Optionally, the Cyclocarya paliurus diseases include Cyclocarya paliurus damping-off and black leaf blight.

[0018] The present invention discloses the following technical effects:

[0019] The present invention combines Bacillus pumilus CICC 22096 and Bacillus subtilis ACCC 64091 and loads the biochar formed by treating Flammulina velutipes residue at a specific temperature. This can provide a good load carrier and microenvironmental conditions for the survival and reproduction of the two bacterial strains and the nutrients they need. The network structure of the biochar with coexistence of large pores and small gaps can enrich nutrients and bacterial strains in the structure at a closer distance, providing convenient conditions for the rapid growth and reproduction of the bacterial strains and the dominant bacterial species. Furthermore, after application to the soil, it can more efficiently provide plants with rich nutrients, which are convenient for the plants to quickly absorb and utilize, thereby laying a foundation for improving plant disease resistance through the absorption and utilization of resistance components in the bacterial strain metabolites.

[0020] By screening bacterial strain compounding conditions and optimizing microbial fertilizer composition and process conditions, this invention produces a microbial fertilizer that significantly improves the resistance of Cyclocarya paliurus to damping-off and black leaf blight. This invention provides a new method and direction for providing highly disease-resistant Cyclocarya paliurus to the market. DETAILED DESCRIPTION

[0021] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0022] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. The intermediate value within any stated value or stated range, and each smaller range between any other stated value or intermediate value within the stated range, is also encompassed within the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0023] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0024] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be illustrative only.

[0025] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0026] The most essential and significant feature of the microbial fertilizer involved in the present invention is that it is a living, active fertilizer. Microbial fertilizer is a biological fertilizer made from one or several beneficial microorganisms through industrial culture and fermentation. Microbial fertilizers are usually divided into two categories: one is that the life activities of the microorganisms contained therein increase the supply of plant nutrients, resulting in improved plant nutritional status and thus increased yields, and its representative variety is microbial fertilizer; the other is microbial fertilizer in a broad sense. Although it also increases crop yields through the life activities of the microorganisms contained therein, it is not limited to improving the supply level of plant nutrients, but also includes the secondary metabolites they produce, such as the stimulating effect of hormones on plants. The main functions of microbial fertilizers are: promoting the absorption and utilization of nutrients by plants, or being able to resist the pathogenic effects of certain pathogenic microorganisms, reducing diseases and pests, and thus improving crop yield and quality.

[0027] The biochar (English: Biochar) involved in the present invention is a type of charcoal used as a soil conditioner, which can help plants grow and can be used for agricultural purposes as well as carbon collection and storage. It is different from traditional charcoal generally used for fuel. Enoki mushroom residue refers to the part remaining after the Enoki mushrooms are picked, including mycelium and inedible parts. Although these wastes are no longer used as food, they contain rich nutrients and have high utilization value. The organic components of Enoki mushroom residue mainly include carbohydrates, proteins, fats, minerals and vitamins. The biochar prepared by carbonizing Enoki mushroom residue in the present invention can be used as a carrier for loading bacterial strains or components of bacterial strain metabolism, providing excellent conditions for the growth and reproduction of bacterial strains.

[0028] The prior art has disclosed a variety of single strains as biocontrol strains for Cyclocarya paliurus disease resistance. More specific implementation methods include: The prior art discloses a Streptomyces biocontrol strain, classified and designated Streptomyces sp. QJK-26, deposited with the China Center for Type Culture Collection, with a deposit number of CCTCC NO: M2018434, and a deposit date of July 2, 2018. Also disclosed is the use of this Streptomyces biocontrol strain in controlling Cyclocarya paliurus leaf black blight, caused by Phomopsis eucommiae. Another example: The prior art discloses a Bacillus biocontrol strain, classified and designated Bacillus sp. QJK-3, deposited with the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, China, with a deposit number of CCTCC NO: M2019377, and a deposit date of May 20, 2019. Also disclosed is the application of the Bacillus biocontrol strain in preventing and controlling Cyclocarya paliurus leaf black blight, which is caused by Phomopsis eucommia.

[0029] However, existing technologies typically use specific strains for the prevention and treatment of Cyclocarya paliurus leaf black blight. There are few reports on the use of composite strains or other single strains for the prevention and treatment of Cyclocarya paliurus leaf black blight. In particular, there are no reports on single or composite strains that can simultaneously prevent and treat both damping-off and post-transplant leaf black blight in Cyclocarya paliurus. Furthermore, the discovery of biocontrol strains is often random. Therefore, in order to meet the market's high demand for both quantity and quality of Cyclocarya paliurus, it is essential to identify composite bacterial agents with resistance to both damping-off and leaf black blight from existing strains in the prior art.

[0030] The present invention relates to the strain:

[0031] Bacillus pumilus (CICC 22096) was purchased from the China Industrial Microbiology Culture Collection Center.

[0032] Bacillus sp., with the deposit number ACCC 64091, was purchased from the China Agricultural Microbiological Culture Collection Center.

[0033] The public can obtain the corresponding strains from the above platforms.

[0034] Antagonism experiment between two strains:

[0035] After activating Bacillus pumilus with nutrient agar medium, the nutrient broth medium was inoculated, and then cultured at 37° C. and 200 rpm for 3 days with shaking. The fermentation liquid was collected and freeze-dried to obtain Bacillus pumilus powder.

[0036] The prepared Bacillus pumilus powder was inoculated onto a nutrient agar medium and incubated at 37°C for 3 days. The Bacillus pumilus on the nutrient agar medium was aseptically removed and then inoculated with Bacillus ACCC 64091. The culture was continued for 48 hours. Simultaneously with the inoculation of Bacillus ACCC 64091, an equal amount of Bacillus ACCC 64091 was inoculated onto a new nutrient agar medium. The culture was incubated for 48 hours and the culture results were observed. The results showed that Bacillus ACCC 64091 grew well in both media, indicating that the two bacteria do not compete with each other and can coexist in the same culture environment.

[0037] Example 1 Optimizing the compound ratio of Bacillus pumilus and Bacillus

[0038] The mixture was prepared according to the ratio shown in Table 1 below, and the number of viable bacteria was determined by plate colony counting method (culture conditions: culture on nutrient agar medium at 37°C for 3 days).

[0039] Table 1 Optimized strain compounding conditions and results

[0040] From the experimental data in Table 1, it can be seen that when Bacillus pumilus powder and Bacillus powder were compounded at a mass ratio of 3:5, the number of viable bacteria was the highest. Therefore, this ratio was used in subsequent experiments.

[0041] Example 2

[0042] A microbial fertilizer comprises the following components in parts by weight: 30 parts of Bacillus pumilus powder, 50 parts of Bacillus powder, and 50 parts of Flammulina velutipes residue biochar. The above components are directly mixed evenly to obtain the microbial fertilizer.

[0043] The preparation of Bacillus pumilus powder refers to the relevant method in the above-mentioned "Verification of the Antagonism Experiment between Two Strains".

[0044] The preparation method of Bacillus powder is the same as that of Bacillus pumilus powder.

[0045] Preparation method of Enoki mushroom residue biochar: select pollution-free and mold-free Enoki mushroom residue, expose it to the sun to kill the remaining bacteria, then pyrolyze the Enoki mushroom residue at 300-400℃ (400℃ was selected in this experiment) for 30 minutes, and then reduce the temperature to 100-200℃ (200℃ was selected in this experiment) for 1 hour to obtain mushroom residue-based biochar.

[0046] Example 3

[0047] Compared with Example 2, the only difference is that the amount of Flammulina velutipes residue biochar added is 30 parts. The other method steps are the same.

[0048] Example 4

[0049] Compared with Example 2, the only difference is that the amount of Flammulina velutipes residue biochar added is 75 parts. The other method steps are the same.

[0050] Comparative Example 1

[0051] Compared with Example 2, the only difference is that Bacillus pumilus powder is not included. Other method steps are the same.

[0052] Comparative Example 2

[0053] Compared with Example 2, the only difference is that Bacillus powder is not included. Other method steps are the same.

[0054] Comparative Example 3

[0055] Compared with Example 2, the only difference is that Bacillus pumilus powder and Bacillus powder are not included. Other method steps are the same.

[0056] Comparative Example 4

[0057] Compared with Example 2, the only difference is that: 30 parts of Bacillus pumilus powder, 40 parts of Bacillus powder and 50 parts of Flammulina velutipes residue biochar are used. Other steps are the same.

[0058] Comparative Example 5

[0059] Compared with Example 2, the only difference is that 10 parts of Bacillus pumilus powder, 20 parts of Bacillus powder and 50 parts of Flammulina velutipes residue biochar are used. Other steps are the same.

[0060] Comparative Example 6

[0061] Compared with Example 2, the only difference is that 20 parts of Bacillus pumilus powder, 50 parts of Bacillus powder and 50 parts of Flammulina velutipes residue biochar are used. Other steps are the same.

[0062] Comparative Example 7

[0063] Compared to Example 2, the only difference is the preparation method of Flammulina velutipes residue biochar. Specifically, pollution-free and mold-free Flammulina velutipes residue is used, which is then exposed to the sun to kill any remaining bacteria. The Flammulina velutipes are then pyrolyzed at 400°C for 90 minutes to produce the mushroom residue-based biochar. All other steps are the same.

[0064] Comparative Example 8

[0065] Compared to Example 2, the only difference is the preparation method of Flammulina velutipes residue biochar. Specifically, pollution-free and mold-free Flammulina velutipes residue is used, which is then exposed to the sun to kill any remaining bacteria. The Flammulina velutipes are then pyrolyzed at 200°C for 90 minutes to produce the mushroom residue-based biochar. All other steps remain the same.

[0066] Comparative Example 9

[0067] Compared with Example 2, the only difference is that the amount of biochar containing Flammulina velutipes residue is 20 parts. The other steps are the same.

[0068] Comparative Example 10

[0069] Compared with Example 2, the only difference is that the amount of biochar containing Flammulina velutipes residue is 85 parts. The other steps are the same.

[0070] Application example: A cultivation method for preventing and controlling Cyclocarya paliurus diseases using microbial fertilizer

[0071] The above cultivation method mainly utilizes the microbial fertilizer prepared in Examples 2-4 and Comparative Examples 1-10 to treat Cyclocarya paliurus, specifically as follows:

[0072] A nursery of the Cyclocarya paliurus planting base in Siduping Township, Zhangjiajie was selected to verify the effects of the microbial fertilizers prepared in Examples 2-4 and Comparative Examples 1-10.

[0073] The nursery was randomly divided into 14 groups, and the planting conditions, growth status, management level and cultivation quantity of the Cyclocarya paliurus in each group were consistent. Among them, group 1 was a blank control group, that is, the microbial fertilizers of the embodiments of the present invention and the comparative examples were not used; groups 2-4 were experimental groups, that is, the microbial fertilizers prepared by embodiments 2-4 of the present invention were used respectively; groups 5-14 were control groups, that is, the microbial fertilizers prepared by comparative examples 1-10 of the present invention were applied respectively. When the Cyclocarya paliurus grew 2-3 leaves, the root application of microbial fertilizer began (after dissolving in water and applying to the roots), the blank control used the same amount of water, and 6 kg of microbial fertilizer was used per mu. The observation was continued until before the Cyclocarya paliurus was transplanted, and the incidence of damping-off disease during the observation period was counted. The results are shown in Table 2.

[0074] Table 2 Statistical results of the incidence of damping-off disease of Populus paliurus in the nursery

[0075]

[0076] The incidence of damping-off disease (%) = the number of plants with damping-off disease in each group / the total number of plants in each group × 100, where the total number of plants in each group is 100.

[0077] The above results show that the use of the microbial fertilizer prepared in Examples 2-4 of the present invention can almost eliminate the occurrence of damping-off disease in the nursery period of Cyclocarya paliurus, and the realization of this effect mainly depends on the synergistic effect between Bacillus pumilus, Bacillus subtilis and Flammulina velutipes residue biochar. If the composition of the strains is changed (such as Comparative Examples 1-3) or the ratio of the strain powder dosage is changed (such as Comparative Examples 4-6), the synergistic effect will be affected. In addition, the generation conditions and dosage of Flammulina velutipes residue biochar also directly affect the effect of the composite strain (such as Comparative Examples 7-10). The Flammulina velutipes residue is first treated at high temperature to form a large-pore network structure, and then the temperature is lowered for continuous carbonization, which can maintain the stability of the biochar. It proves that there are a large number of large-pore network structures, and the mechanical strength and stability are also good. At the same time, some structures with smaller gaps can be formed. The gaps of such sizes are staggered. While providing oxygen conditions for the two strains to recombine, the macromolecules degraded by the two strains, such as protein molecules, can also attach to the pores, providing the two strains with absorbable nutrients more closely, which is conducive to the rapid formation of dominant bacterial communities in a limited space. Then, when applied to the soil, it makes full use of the favorable conditions for the aggregation of dominant bacterial communities and nutrients, lays the foundation for the plants to quickly absorb and utilize nutrients, and significantly inhibits the occurrence of green poplar blight during the nursery period through the absorption and utilization of resistance components in nutrients.

[0078] One year after the Cyclocarya paliurus was transplanted in the above nursery, the incidence of black leaf blight was counted within two months from the time when tender leaves began to grow. The results are shown in Table 3 below.

[0079] Table 3 Statistical results of the incidence of black blight on transplanted Cyclocarya paliurus seedlings

[0080]

[0081] The incidence of black leaf blight (%) = the number of plants with black leaf blight in each group / the total number of transplanted plants in each group × 100, where the total number of transplanted plants in each group is 60.

[0082] In order to further directly verify the biocontrol effect of the microbial fertilizer prepared by the present invention, the biocontrol effect was verified by a pathogen inoculation experiment.

[0083] The black leaf blight pathogen was inoculated into PDA medium and cultured at room temperature. Conidia were collected and prepared into 1×10 6cfu / mL spore suspension. Soil was collected from a Cyclocarya paliurus nursery, air-dried for three days, crushed, and sieved. A spore suspension of the pathogen causing leaf blight was then evenly sprayed on the soil. After thorough mixing, the infected soil was prepared. The infected soil was then divided into identical pots, filling two-thirds of the pot volume. One healthy, one-year-old Cyclocarya paliurus seedling was transplanted into each pot and cultured in a greenhouse. The Cyclocarya paliurus plants in the pots were randomly divided into 14 groups, each containing five pots. Fifteen days after transplanting, the plants were treated as follows: Group 1 served as a blank control group, treated with an equal amount of sterile water instead of the microbial fertilizers prepared in Examples 2-4 of the present invention (the experimental group microbial fertilizers were dissolved in water). Groups 5-14 served as controls, treated with the microbial fertilizers prepared in Comparative Examples 1-10 of the present invention (the control group microbial fertilizers were dissolved in water). For each Cyclocarya paliurus plant, 1g of microbial fertilizer was dissolved in 100mL of water and sprayed evenly on the surface of Cyclocarya paliurus leaves with a spray bottle. After 30 days of treatment, the disease was investigated.

[0084] Disease grade standards: Grade 0: No leaf blight symptoms; Grade 1: 1-25% of leaves show leaf blight symptoms; Grade 2: 26-50% of leaves show leaf blight symptoms; Grade 3: 51-75% of leaves show leaf blight symptoms; Grade 4: 76% or more of leaves show leaf blight symptoms. Disease index = Σ[number of diseased leaves at each grade × relative disease grade value at each grade] / [total number of leaves surveyed] × highest disease grade value. Control efficacy (%) = (control disease index – treatment disease index) / control disease index × 100.

[0085] Table 4 Survey results of black blight on Cyclocarya paliurus leaves in greenhouse pots

[0086]

[0087] The above results show that the microbial fertilizers prepared in Examples 2-4 of the present invention have significantly improved the function of resisting leaf black blight, and the prevention efficiency can reach more than 90%, which is significantly improved compared with the 60-70% biological prevention efficiency of the prior art.

[0088] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.

Claims

1. A cultivation method for preventing and controlling Cyclocarya paliurus diseases using microbial fertilizer, characterized in that: The method comprises the steps of treating Cyclocarya paliurus with microbial fertilizer; wherein the microbial fertilizer comprises the following components in parts by weight: Bacillus pumilus ( Bacillus pumilus ) 30 parts of bacterial powder, Bacillus ( Bacillus sp. ) 50 parts of bacterial powder and 30-75 parts of Flammulina velutipes residue biochar; the Bacillus pumilus is Bacillus pumilus CICC 22096, and the Bacillus is Bacillus subtilis ACCC64091; The steps for preparing the Flammulina velutipes residue biochar include: pyrolyzing the Flammulina velutipes residue at a high temperature of 300-400° C. for 30 minutes, and then reducing the high temperature to 100-200° C. for 1 hour to obtain mushroom residue-based biochar.

2. The cultivation method according to claim 1, characterized in that The method of treating Cyclocarya paliurus with microbial fertilizer includes: applying the microbial fertilizer to the roots of Cyclocarya paliurus in the nursery stage or spraying the microbial fertilizer on the leaves of Cyclocarya paliurus transplanted seedlings.

3. The cultivation method according to claim 1, characterized in that 5-7 kg of the microbial fertilizer is used per mu of Cyclocarya paliurus seedlings.

4. A microbial fertilizer for preventing and controlling Cyclocarya paliurus diseases, characterized in that: The invention comprises the following components in parts by weight: 30 parts of Bacillus pumilus powder, 50 parts of Bacillus subtilis powder and 30-75 parts of Flammulina velutipes residue biochar; The Bacillus pumilus is Bacillus pumilus CICC 22096, and the Bacillus is Bacillus subtilis ACCC64091.

5. Use of the microbial fertilizer as claimed in claim 4 in preventing and treating Cyclocarya paliurus diseases.

6. The use according to claim 5, characterized in that The diseases of Cyclocarya paliurus include Cyclocarya paliurus damping-off and black leaf blight.

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