Cultivation method for preventing and treating diseases of cyclocarya paliurus by using microbial fertilizer

Abstract

The invention discloses a cultivation method for preventing and treating diseases of cyclocarya paliurus by using bacterial manure, and belongs to the field of prevention and treatment of diseases in agriculture and forestry. The cultivation method comprises the step of treating the cyclocarya paliurus by using a microbial fertilizer, the microbial fertilizer is prepared from 30 parts of bacillus pumilus powder, 50 parts of bacillus powder and 30 to 75 parts of flammulina velutipes residue charcoal; the bacillus pumilus is bacillus pumilus CICC 22096, and the bacillus is bacillus subtilis ACCC 64091. The bacillus pumilus is bacillus pumilus CICC 22096, and the bacillus is bacillus subtilis ACCC 64091. After the bacillus pumilus CICC 22096 and the bacillus ACCC 64091 are compounded, the flammulina velutipes mushroom dreg charcoal is used as a carrier, so that excellent conditions can be provided for growth and reproduction of strains and loading of metabolites, rapid formation of dominant flora is promoted, resistant components in the metabolites are supplied to plants in a more concentrated manner, rapid absorption and utilization of the plants are promoted, and the yield of the plants is increased. A foundation is laid for improving the 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 is a deciduous tree of the genus Cyclocarya paliurus in the family Juglandaceae. Its fruits are like strings of copper coins and are quite ornamental. It can be used as an ornamental tree species and timber species for landscaping. Its young leaves have medicinal value such as lowering blood pressure, blood sugar, and blood lipids, enhancing human immunity, anti-oxidation, and anti-aging. Its wood is also a high-quality furniture material. As Cyclocarya paliurus has great application value in many fields, the market demand has also increased. Therefore, if we want to ensure the continuous market supply of Cyclocarya paliurus, we need to ensure that the quality of Cyclocarya paliurus is not reduced every year.

[0003] However, the seedling stage of Cyclocarya paliurus is very prone to diseases, 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 transplanting from nurseries. 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 disinfects the nursery before sowing, and uses chemical agents such as methyl thiophanate, thiophanate-methyl, carbendazim and other drugs for prevention and control after damping-off or black leaf blight occurs. Although chemical control is effective, long-term use causes environmental pollution and resistance of pathogenic microorganisms. Therefore, it is very important to find new methods for preventing and controlling Cyclocarya paliurus.

[0004] Microbial fertilizer is a new type of fertilizer biological product that uses the life activities of active microorganisms to enable crops to obtain the nutrients they need. It is a type of fertilizer in agricultural production (also known as the third generation of fertilizer). Microbial fertilizer has been applied to agricultural disease prevention and control, and the effect is remarkable. At present, there are reports on the use of single bacterial agents to prevent and control diseases of Cyclocarya paliurus, but the prevention effect of a single bacterial agent is generally 60%-70%, and it is difficult to restore the microecological structure. Not all strains have a biocontrol effect, and the synergistic effects of different strains and plants vary greatly. As the market demand for the quantity and quality of Cyclocarya paliurus increases, seeking a single bacterial agent or a compound bacterial agent with better biocontrol effects is crucial to solving the problem of poor disease control of Cyclocarya paliurus. Summary of the invention

[0005] The invention aims to provide a cultivation method for preventing and controlling Cyclocarya paliurus diseases by utilizing microbial fertilizer, so as to solve the problems existing in the above-mentioned prior art. After compounding Bacillus pumilus CICC 22096 and Bacillus subtilis ACCC 64091, Flammulina velutipes residue biochar is used as a carrier, which can provide superior conditions for the growth and reproduction of strains and the load of metabolites, promote the rapid formation of dominant bacterial flora, 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 the disease resistance of plants.

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

[0007] The invention provides a cultivation method for preventing and controlling Cyclocarya paliurus diseases by using microbial fertilizer, comprising the step of treating Cyclocarya paliurus by using the microbial fertilizer; wherein 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 in the nursery stage 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 the Cyclocarya paliurus seedlings in the nursery period or transplanted from the nursery. If it is for the prevention and treatment of black blight of adult leaves, the dosage can be appropriately adjusted according to demand.

[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 subtilis 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 and culturing 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 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 forms a biochar load by compounding Bacillus pumilus CICC 22096 and Bacillus subtilis ACCC 64091 and treating Flammulina velutipes residue at a specific temperature, so as to provide a good load carrier and microenvironment conditions for the survival and reproduction of the two strains and the required nutrients. The reticular structure of the biochar with coexistence of macropores and microvoids can enrich the nutrients and strains in the structure at a closer distance, thereby providing convenient conditions for the rapid growth and reproduction of the strains and the dominant strains. After being applied to the soil, the biochar can more efficiently provide rich nutrients for plants, which are convenient for the plants to quickly absorb and utilize, thereby laying a foundation for improving the disease resistance of plants through the absorption and utilization of resistance components in the strain metabolites.

[0020] The present invention prepares a microbial fertilizer that can significantly improve the resistance of Cyclocarya paliurus to damping-off and black leaf blight by screening bacterial strain compounding conditions and optimizing microbial fertilizer composition and process conditions. The present 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 should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

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

[0023] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation 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 a conflict with any incorporated document, the content 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 of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary 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 of one or several beneficial microorganisms through industrial culture and fermentation. Microbial fertilizers are usually divided into two categories: one is that through the life activities of the microorganisms contained therein, the supply of plant nutrients is increased, resulting in the improvement of plant nutritional status and thus increasing yields. Its representative variety is bacterial fertilizer; the other is a broad microbial fertilizer. 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 insect pests, and thus improving crop yields and quality.

[0027] The biochar (English: Biochar) involved in the present invention is a kind 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 as fuel. Flammulina velutipes residue refers to the part remaining after the Flammulina velutipes is 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 Flammulina velutipes residue mainly include carbohydrates, proteins, fats, minerals and vitamins. The biochar prepared by carbonizing the Flammulina velutipes residue in the present invention can be used as a carrier for loading strains or components of strain metabolism, providing excellent conditions for the growth and reproduction of strains.

[0028] In the prior art, a variety of single strains have been disclosed as biocontrol strains for the disease resistance of Cyclocarya paliurus. More specific implementation methods are as follows: The prior art discloses a biocontrol strain of Streptomyces, which is classified and named Streptomyces sp. QJK-26, deposited in 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 the biocontrol strain of Streptomyces in the prevention and treatment of black blight of Cyclocarya paliurus leaves, which is caused by Phomopsis eucommiae. Another example: The prior art discloses a biocontrol strain of Bacillus sp. QJK-3, which is deposited in the China Center for Type Culture Collection, referred to as CCTCC, with a deposit address of 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 treating Cyclocarya paliurus leaf black blight, wherein the Cyclocarya paliurus leaf black blight is caused by Phomopsis eucommiae.

[0029] However, the prior art uses specific strains for the prevention and treatment of Cyclocarya paliurus leaf black blight, and there are few reports on the use of composite strains or other single strains in the prevention and treatment of Cyclocarya paliurus leaf black blight, especially single strains or composite strains for the prevention and treatment of Cyclocarya paliurus damping-off in the nursery period and leaf black blight after transplanting from the nursery, and the discovery of biological control strains is random. Therefore, in order to meet the market's large demand for the quantity and quality of Cyclocarya paliurus, it is very necessary to mine composite bacterial agents with resistance to Cyclocarya paliurus damping-off and leaf black blight from existing strains in the prior art.

[0030] The present invention relates to the strain:

[0031] Bacillus pumilus, with the deposit number of CICC 22096, was purchased from China Industrial Microbiological Culture Collection Center;

[0032] Bacillus sp., with the deposit number ACCC 64091, was purchased from 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, inoculate nutrient broth medium, then culture under shaking conditions of 37° C. and 200 rpm for 3 days, collect the fermentation liquid, and freeze-dry to obtain Bacillus pumilus powder.

[0036] The prepared Bacillus pumilus powder was inoculated into the nutrient agar medium, and the mixture was cultured at 37°C for 3 days. The Bacillus pumilus on the nutrient agar medium was aseptically removed, and Bacillus ACCC 64091 was inoculated again, and the culture was continued for 48 hours. While inoculating Bacillus ACCC 64091, an equal amount of Bacillus ACCC 64091 was inoculated into the new nutrient agar medium, and the mixture was cultured for 48 hours, and the culture results were observed. The results showed that Bacillus ACCC 64091 grew well in both culture media, indicating that the two bacteria were not antagonistic and could coexist in the same culture environment.

[0037] Example 1 Optimizing the compounding 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 the plate colony counting method (culture conditions: culture on nutrient agar medium at 37°C for 3 days).

[0039] Table 1 Compounding conditions and results of optimized strains

[0040] It can be seen from the experimental data in Table 1 that when Bacillus pumilus powder and Bacillus powder were compounded in 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 the preparation method of Bacillus pumilus powder.

[0045] Preparation method of Flammulina velutipes residue biochar: select pollution-free and mold-free Flammulina velutipes residue, expose it to the sun to kill the remaining bacteria, then pyrolyze the Flammulina velutipes residue at 300-400℃ (400℃ was selected in this experiment) for 30min, then reduce the temperature to 100-200℃ (200℃ was selected in this experiment) for 1h 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. The 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. The other steps of the method are the same.

[0058] Comparative Example 5

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

[0062] Comparative Example 7

[0063] Compared with Example 2, the only difference is that the preparation method of Flammulina velutipes residue biochar is different, that is, pollution-free and mold-free Flammulina velutipes residue is selected, exposed to the sun to kill the residual bacteria, and then the Flammulina velutipes is pyrolyzed at 400° C. for 90 minutes to obtain mushroom residue-based biochar. The other method steps are the same.

[0064] Comparative Example 8

[0065] Compared with Example 2, the only difference is that the preparation method of Flammulina velutipes residue biochar is different, that is, pollution-free and mold-free Flammulina velutipes residue is selected, exposed to the sun to kill the residual bacteria, and then the Flammulina velutipes is pyrolyzed at a high temperature of 200° C. for 90 minutes to obtain mushroom residue-based biochar. The other method steps are the same.

[0066] Comparative Example 9

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

[0068] Comparative Example 10

[0069] Compared with Example 2, the only difference is that the amount of Flammulina velutipes residue biochar is 85 parts. The other method 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 uses the microbial fertilizer prepared in Examples 2-4 and Comparative Examples 1-10 to treat Cyclocarya paliurus, 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 the above 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, and 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 grows 2-3 leaves, the root application of microbial fertilizers (root application after dissolving in water) was started, and the blank control used an equal amount of water, and 6 kg of microbial fertilizer was used per mu. The observation was continued until before the transplanting of the Cyclocarya paliurus, and the incidence of damping-off during the observation period was counted. The results are shown in Table 2.

[0074] Table 2 Statistical results of the incidence of Rhizoctonia solani 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 makes it possible to prevent 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 and Bacillus 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 use effect of its composite strains (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 with good mechanical strength and stability, and some structures with smaller gaps can also be formed. The interlacing of large and small gaps can not only provide oxygen conditions for the two strains to recombine, but also enable the macromolecules degraded by the two strains, such as protein molecules, to attach to the pores, and provide the two strains with absorbable nutrients more closely, which is conducive to the rapid formation of dominant bacterial communities in a limited space. When applied to the soil, it fully utilizes the favorable conditions for the aggregation of dominant bacterial communities and nutrients, lays the foundation for 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 leaf black blight was counted within 2 months from the beginning of the growth of tender leaves. 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, and the total number of transplanted plants in each group is 60 plants.

[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. A certain Cyclocarya paliurus nursery was selected, soil was taken, dried for 3 days, crushed, sieved, and then evenly sprayed with a spore suspension of leaf black blight pathogen, and mixed thoroughly to make a contaminated soil. Then, the contaminated soil was divided into the same cultivation pots, and the amount of soil was 2 / 3 of the total volume of the cultivation pots. One healthy annual Cyclocarya paliurus seedling was transplanted into each pot and cultured in a greenhouse. The Cyclocarya paliurus in the cultivation pots were randomly divided into 14 groups, each with 5 pots. After 15 days of transplanting, they were treated according to the following method: Group 1 was a blank control group, that is, the microbial fertilizer of the embodiment of the present invention and the comparative example was not used, and the same amount of sterile water was used for treatment; Groups 2-4 were experimental groups, that is, the microbial fertilizer prepared by Examples 2-4 of the present invention was used for treatment (the microbial fertilizer of the experimental group was dissolved in water), and Groups 5-14 were control groups, that is, the microbial fertilizer prepared by Comparative Examples 1-10 of the present invention was used for treatment (the microbial fertilizer of the control group was dissolved in water). Use 1g of microbial fertilizer for each Cyclocarya paliurus, dissolve 100mL in water, and spray it evenly on the surface of Cyclocarya paliurus leaves with a spray bottle. After 30 days of treatment, investigate the condition.

[0084] Disease level standard: Level 0, no leaf black blight symptoms; Level 1, 1-25% of leaves show leaf black blight symptoms; Level 2, 26-50% of leaves show leaf black blight symptoms; Level 3, 51-75% of leaves show leaf black blight symptoms; More than 76% of leaves show leaf black blight symptoms. Disease index = Σ[number of diseased leaves at each level × relative disease level value at each level] / [total number of leaves surveyed] × highest disease level value. Prevention effect (%) = (control disease index - treatment disease index) / control disease index × 100.

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

[0086]

[0087] It can be seen from the above results that the microbial fertilizers prepared in Examples 2-4 of the present invention have the function of significantly improving resistance to 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 only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined 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: 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.

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, 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. A method for preparing the microbial fertilizer according to claim 4, characterized in that: The method comprises uniformly mixing the components in the formula.

6. The preparation method according to claim 5, characterized in that: The Bacillus pumilus powder is obtained by activating Bacillus pumilus CICC 22096, fermenting and culturing it in a nutrient broth medium at 35-37° C. for 3 days, collecting the fermentation liquid, and freeze-drying it; 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.

7. The preparation method according to claim 5, characterized in that: 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.

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

9. The use according to claim 8, characterized in that The diseases of Cyclocarya paliurus include Cyclocarya paliurus damping-off and black leaf blight.

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