Citrus trimmed branch compost complex microbial inoculant as well as preparation method and application thereof

Abstract

The invention relates to a new technology of cyclic ecological agriculture and biomass resource utilization, in particular to a citrus trimmed branch compost complex microbial inoculant as well as a preparation method and application thereof. The composite microbial agent is formed by compounding bacteria and fungi, the bacteria comprise Delftia delfinii, bacillus velezensis, bacillus subtilis and achromobacter, the fungi comprise alternaria alternata, penicillium oxalicum, candida mycoderma bacteria, humicola insolens and aspergillus niger, and the total viable count of the microbial agent is not less than 8.3 lg cfu / mL. The trimmed citrus branch compost composite microbial agent provided by the invention overcomes the defect that the existing microbial agent has a poor compost effect on trimmed citrus branches through the synergistic effect of the strains.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of agriculture and biological resources, in particular to a new technology of circular ecological agriculture and utilization of biomass resources, and specifically to a citrus pruning branch composting complex microbial agent, a preparation method and application thereof. BACKGROUND

[0002] Citrus, as one of the most important economic crops in the world, plays a crucial role in agricultural production. It not only provides humans with abundant vitamin C and other nutrients, but also plays a key role in agricultural economy, food processing and international trade. The global citrus planting area is vast, especially in subtropical and tropical regions, and the citrus industry has become a pillar industry in many countries and regions. China, as one of the main citrus producing countries, ranks among the world's top in terms of citrus planting area and yield, and the citrus industry is of great significance to farmers' income increase, rural economic development and agricultural structure adjustment. However, with the rapid development of the citrus industry, related production management problems have become increasingly prominent, among which the disposal of citrus pruning branches has become a challenge that cannot be ignored.

[0003] In the annual management of citrus, pruning is a crucial technical measure. Through pruning, the tree structure can be adjusted, the ventilation and light conditions can be improved, the occurrence of diseases and pests can be reduced, and the fruit quality and yield can be improved. However, the amount of branch waste generated during pruning is huge. According to statistics, the amount of branch waste generated per mu of citrus garden per year through pruning can reach several hundred kilograms to one ton, and the specific amount varies with variety, tree age, cultivation mode and pruning intensity. These pruning branches are traditionally randomly piled up or burned, which not only occupies land resources, but also may cause fire hazards, and the smoke and harmful gases generated during the burning process pollute the atmospheric environment, which is contrary to the concept of green agriculture and sustainable development. How to efficiently and environmentally dispose of these organic wastes has become a problem that needs to be solved in the sustainable development of the citrus industry.

[0004] Turning citrus pruning branches into organic fertilizer through composting is a very potential resource utilization way. Composting is a process of decomposing and transforming organic matter into stable humus through the action of microorganisms, and its products can be used to improve soil structure, increase soil fertility, and reduce the use of chemical fertilizers. Domestic and foreign researches on the composting of agricultural wastes have accumulated rich achievements. For example, composting microbial agents for rice straw, corn straw and other crop residues have been relatively mature, but there is still room for improvement in the treatment of fruit tree branches such as citrus pruning branches. This is mainly due to the higher degree of lignification of fruit tree branches, and the possible presence of certain secondary metabolites (such as phenolic compounds in citrus), which inhibit microbial activity. Therefore, developing a special composting complex microbial agent for citrus pruning branches to accelerate the degradation and composting of citrus pruning branches has become a research hotspot for the resource utilization of citrus waste. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a compound microbial agent for citrus pruning composting, its preparation method, and its application. The microbial strains used in this application overcome the shortcomings of existing microbial agents in achieving poor composting effects on citrus pruning branches through synergistic effects.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: A compound microbial agent for citrus pruning compost, comprising bacteria and fungi; wherein the bacteria include Delftobacterium (…). Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromobacter ); The fungi include Alternaria ( Nimbya alternantherae ), Penicillium oxalate ( Penicillium oxalicum ), Candida albicans ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger ( Aspergillus niger ).

[0007] Preferably, the number of live bacteria in the composting compound microbial agent is not less than 8.3 1 g CFU / mL.

[0008] Preferably, the viable bacterial count in the composting compound microbial agent is not less than 4.4 1 g CFU / mL, and the viable fungal count is not less than 3.9 1 g CFU / mL.

[0009] Preferably, the bacteria in the composting compound microbial agent are Delftobacterium (…). Delftia tsuruhatensis The viable count of *Bacillus belyssus* was 2.0–3.0 lg cfu / mL. Bacillus velezensis The viable count of Bacillus subtilis was 2.5~3.51g CFU / mL. Bacillus subtilis The viable bacterial count was 2.5–3.0 lg CFU / mL and Achromobacterium ( Achromobacter The viable bacterial count was 1.0~1.5 lg cfu / mL; The fungi in the composting compound microbial agent include Alternaria ( Nimbya alternantherae The viable count of Penicillium oxalate was 0.5~1.2 lg cfu / mL. Penicilliumoxalicum The viable cell count was 1.0~1.5 lg CFU / mL, and the viable cell count of Candida albicans was 1.0~1.5 lg CFU / mL. Candida The viable count of ) was 0.75~1.5 lg cfu / mL, and that of specific saprophytic molds ( Vanrija Humicola The viable count of ) was 1.25~2.0 lg cfu / mL and Aspergillus niger (Aspergillusniger The viable count was 0.5~1.5 lg cfu / mL.

[0010] A method for preparing a compound microbial agent for citrus pruning compost includes the following steps: S1, Bacterium delavayi ( Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromo bacter Alternaria ( Nimbya alternantherae ), Penicillium oxalate ( Penicillium oxalicum ), Candida albicans ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger ( Aspergillus niger The strains were isolated and purified by incubation at 37°C for 48 hours, and the purified strains were inoculated onto LB slant agar. S2, Bacterium deltaeoides ( Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromo bacter The strains were inoculated into 300 mL of LB liquid medium and cultured in a shaker at 37℃ and 180 r / min for 1-2 days until the viable count in the bacterial solution was not less than 4.4 lg cfu / mL. S3. Select Alternaria from the purified strains ( Nimbya alternantherae ), Penicillium oxalate ( Penicillium oxalicum ), Candida albicans ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger ( Aspergillus niger The mycelia and spores of the mycelia were inoculated into 200 mL of PD medium and cultured at 30℃ and 200 r / min for 5 days to allow the mycelial balls to cover the entire PD liquid medium and the viable count to be no less than 3.9 lg cfu / mL. S4, Add Delft bacteria ( Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromo bacter The culture medium of the bacteria was mixed in equal volume ratio to construct bacterial agent C1; S5, Alternaria ( Nimbya alternantherae ), Penicillium oxalate ( Penicillium oxalicum ), Candida albicans ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger (Aspergillus niger The culture medium of the bacteria was mixed in equal volume ratio to construct bacterial agent C2; S6. Mix the culture media of bacterial agent C1 and bacterial agent C2 in equal volume ratio to construct bacterial agent C3, thus obtaining a compound bacterial agent.

[0011] Preferably, the LB liquid culture medium is prepared as follows: 10g of tryptone, 5g of yeast extract, and 10g of sodium chloride are weighed sequentially and added to a container containing 950ml of deionized water. The container is shaken until the solute is dissolved. Then, 5mol / L sodium hydroxide is added to the container to adjust the pH to 7.4. Deionized water is added to the container to bring the volume to 1L. The container is then placed in a pressure sterilizer and steam-sterilized at 15psi for 20min to obtain the LB liquid culture medium. The preparation method of the LB solid culture medium is as follows: Weigh 10g of tryptone, 5g of yeast extract, and 10g of sodium chloride in sequence, and add them to a container containing 950ml of deionized water. Shake the container until the solute dissolves. Then add 15g of agar powder and 5mol / L sodium hydroxide to the container to adjust the pH value to 7.4. Add deionized water to the container to make up the volume to 1L. Then place the container in a pressure sterilizer and steam sterilize it at 15psi for 20min to obtain LB slant culture medium.

[0012] Preferably, the preparation method of the PD liquid culture medium is as follows: Weigh 200g of potatoes, wash, peel and slice them, put them in a pot, add 1000mL of water, boil for 15min, then filter with gauze, continue to add water to the pot to make the water in the pot reach 1000mL, then add 12g of glucose, and boil until the solid is fully dissolved, and then dispense to obtain PD liquid culture medium; sterilize the dispensed PD culture medium within 1h by high pressure steam moist heat sterilization, at a steam pressure of 0.1 MPa and a temperature of 121 ℃, maintained for 20min.

[0013] An application of a compound microbial agent for fermenting citrus pruning branches in citrus pruning branch composting involves diluting the compound microbial agent after 3-7 days of composting and spraying it evenly at a rate of 5 kg per ton of citrus pruning branches. The dilution ratio of the compound microbial agent is 800-1000 times.

[0014] Preferably, a compound microbial agent is applied to the mixture of citrus pruning branches and manure, wherein the compound microbial agent is applied at a ratio of 5 kg / ton of the total mass of the mixture.

[0015] Preferably, the compound microbial agent is applied to a mixture of shredded citrus pruning branches and manure, wherein the compound microbial agent is applied at a mass ratio of 3 kg / ton of the total mixture. The technical solution provided by this invention can include the following beneficial effects: This invention discloses a compound microbial agent for citrus pruning compost, its preparation method and application. By combining four kinds of bacteria and five kinds of fungi, the synergistic effect between them can effectively improve the degradation effect of citrus pruning branches, significantly shorten the composting cycle and improve the quality of compost. Attached Figure Description

[0016] Figure 1 The graphs show the temperature changes over time in compost bodies under different treatments in Examples 2, 3, 4, and 5 of the present invention, as well as Comparative Examples 1 and 2. Figure 2 The graphs show the changes in moisture content of compost bodies under different treatments in Examples 2, 3, 4, and 5 of the present invention, as well as Comparative Examples 1 and 2. Figure 3 The graphs show the changes in C / N values ​​of compost bodies under different treatments in Examples 2, 3, 4, and 5 of the present invention, as well as Comparative Examples 1 and 2. Figure 4 The bar chart shows the seed germination rate of compost products from different treatments in Examples 2, 3, 4, 5 and Comparative Examples 1 and 2 of the present invention. Detailed Implementation

[0017] To enable those skilled in the art to better understand the technical solutions in this specification, the technical solutions in the embodiments of this specification will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this specification, and not all embodiments. Based on the embodiments in this specification, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of this specification.

[0018] Example 1

[0019] The compound microbial agent was prepared by the following method, and the same method was repeated three times to obtain compound microbial agent 1, compound microbial agent 2 and compound microbial agent 3 respectively; S1, Bacterium delavayi ( Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromobacter Alternaria ( Nimbya alternantherae ), Penicillium oxalate ( Penicilliumoxalicum ), Candida albicans ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger ( Aspergillusniger The strains were isolated and purified by incubation at 37°C for 48 hours, and the purified strains were inoculated onto LB slant agar. S2, Bacterium deltaeoides ( Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromo bacter The strains were inoculated into 300 mL of LB liquid medium and cultured in a shaker at 37℃ and 180 r / min for 1-2 days until the viable count in the bacterial solution was not less than 4.4 lg cfu / mL. S3. Select Alternaria from the purified strains ( Nimbya alternantherae ), Penicillium oxalate ( Penicillium oxalicum ), Candida albicans ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger ( Aspergillus niger The mycelia and spores of the mycelia were inoculated into 200 mL of PD medium and cultured at 30℃ and 200 r / min for 5 days to allow the mycelial balls to cover the entire PD liquid medium and the viable count to be no less than 3.9 lg cfu / mL. S4, Add Delft bacteria ( Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromo bacter The culture medium of the bacteria was mixed in equal volume ratio to construct bacterial agent C1; S5, Alternaria ( Nimbya alternantherae ), Penicillium oxalate ( Penicillium oxalicum ), Candida albicans ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger ( Aspergillus niger The culture medium of the bacteria was mixed in equal volume ratio to construct bacterial agent C2; S6. Mix the culture media of bacterial agent C1 and bacterial agent C2 in equal volume ratio to construct bacterial agent C3, thus obtaining a compound bacterial agent.

[0020] Compound microbial agent 1: Delftobacterium ( Delftia tsuruhatensis The effective viable count was 2.0 lgcfu / mL; Bacillus belyssus ( Bacillus velezensis The effective viable count of Bacillus subtilis was 2.5 lg CFU / mL; Bacillus subtilis The effective viable count was 2.94 lg CFU / mL; Achromobacterium ( Achromo bacter The effective viable count of Alternaria was 1.2 lg CFU / mL; Nimbya alternantherae The effective viable count of ) was 0.97 lg cfu / mL; Penicillium oxalate ( Penicillium oxalicumThe effective viable count of ) was 1.2 lg CFU / mL; Candida albicans ( Candida The effective viable count was 0.79 lg cfu / mL; specific saprophytic molds ( Vanrija Humicola The effective viable count of Aspergillus niger was 1.45 lg cfu / mL; Aspergillus niger The effective viable count was 0.82 lg cfu / mL.

[0021] Compound microbial agent 2: Delft bacillus ( Delftia tsuruhatensis The effective viable count was 2.13 lgcfu / mL; Bacillus belyssus ( Bacillus velezensis The effective viable count of Bacillus subtilis was 2.37 lg CFU / mL; Bacillus subtilis The effective viable count was 2.85 lg CFU / mL; Achromobacterium ( Achromo bacter The effective viable count of Alternaria was 1.32 lg CFU / mL; Nimbya alternantherae The effective viable count of ) was 1.06 lg cfu / mL; Penicillium oxalate ( Penicillium oxalicum The effective viable count of ) was 1.26 lg CFU / mL; Candida albicans ( Candida The effective viable count was 0.93 lg cfu / mL; specific saprophytic molds ( Vanrija Humicola The effective viable count of Aspergillus niger was 1.56 lg cfu / mL; Aspergillus niger The effective viable count was 0.85 lg cfu / mL.

[0022] Compound microbial agent 3: Delft bacterium ( Delftia tsuruhatensis The effective viable count of Bacillus belysin was 2.25 lgcfu / mL; Bacillus velezensis The effective viable count of Bacillus subtilis was 2.66 lg CFU / mL; Bacillus subtilis The effective viable count was 2.84 lg CFU / mL; Achromobacterium ( Achromo bacter The effective viable count of Alternaria was 1.45 lg cfu / mL; Nimbya alternantherae The effective viable count of ) was 1.2 lg cfu / mL; Penicillium oxalate ( Penicillium oxalicum The effective viable count of ) was 1.37 lg CFU / mL; Candida albicans ( Candida The effective viable count of ) was 0.95 lg cfu / mL; specific saprophytic mold ( Vanrija Humicola The effective viable count of Aspergillus niger was 1.62 lg cfu / mL;Aspergillus niger The effective viable count was 0.87 lg cfu / mL.

[0023] Example 2

[0024] In this embodiment, citrus pruning branches were used for composting and decomposition experiments, and indicators such as moisture content, C / N ratio, and seed germination rate were measured. The specific steps are as follows: (1) Take 100 kg of citrus pruning branches, compost them with 200 kg of water for 3-7 days, and use an Elementar vario MACRO carbon and nitrogen analyzer to determine the total carbon (TC) and total nitrogen (TN) content of the air-dried sample that has passed through a 0.15 mm sieve.

[0025] (2) Add 0.5 kg of the compound microbial agent 1 obtained in Example 1 with a dilution ratio of 1000 times to the compost in step (1).

[0026] (3) Record the fermentation temperature on days 0, 3, 7, 14, 21, 28, 35, 42, 49, 56 and 63 of composting, and retain fresh and air-dried samples for determining moisture content and C / N value.

[0027] (4) After composting, weigh 10 g of fresh sample, use deionized water at a fertilizer-to-water ratio of 1:10, shake horizontally at 200 r / min at room temperature, filter, take 5 mL of filtrate, add it to a 9 cm petri dish lined with 2 sheets of filter paper, put 50 Shanghai bok choy seeds evenly, and culture in a dark incubator at 30℃ for 48 h. Calculate the germination rate and repeat 3 times.

[0028] The results showed that during the composting process in this embodiment, easily decomposable organic matter rapidly decomposed under the action of microorganisms, generating a large amount of heat that caused the temperature of the compost pile to rise rapidly, reaching the sanitary standard for harmless composting, which is a maximum temperature greater than 50℃ and maintained for more than 120-240 hours. Subsequently, the temperature of the compost pile began to decrease, and the pile entered a stable maturation process. Finally, the temperature of the compost pile tended to approach the ambient temperature, indicating that this embodiment can complete composting fermentation in 70 days. Figure 1 ), and the water content decreases rapidly ( Figure 2 The C / N ratio decreased significantly. Figure 3 Compost products had no significant effect on the germination of Shanghai bok choy seeds. Figure 4 ).

[0029] Example 3

[0030] This embodiment is the same as the steps in embodiment 2, except that the citrus pruning branches were crushed and 0.3 kg of the compound microbial agent 1 obtained in embodiment 1 with a dilution ratio of 1000 times was added to the compost in step (1).

[0031] The results showed that this embodiment could complete composting fermentation in 70 days. Figure 1 ), and the water content decreases rapidly ( Figure 2 The C / N ratio decreased significantly. Figure 3 Compost products had no significant effect on the germination of Shanghai bok choy seeds. Figure 4 Compared to Example 2, the moisture content and C / N ratio in this example changed significantly with the number of composting days, indicating that the composting was more thorough.

[0032] Example 4

[0033] This embodiment follows the same steps as Embodiment 2, except that 100 kg of pig manure and 100 kg of citrus pruning branches are mixed together.

[0034] The results showed that this embodiment could complete composting fermentation in 63 days. Figure 1 ), and the water content decreases rapidly ( Figure 2 The C / N ratio decreased significantly. Figure 3 Compost products had no significant effect on the germination of Shanghai bok choy seeds. Figure 4 Compared with Examples 2 and 3, the temperature, moisture content, and C / N ratio of the compost pile in this example changed significantly with the number of composting days, indicating that the composting was more thorough.

[0035] Example 5

[0036] This embodiment is the same as the steps in embodiment 3, except that 100 kg of pig manure and 100 kg of citrus pruning branch powder were mixed together, and 0.3 kg of the compound microbial agent 1 obtained in embodiment 1 with a dilution ratio of 1000 times was added to the compost in step (1).

[0037] The results showed that this embodiment could complete composting fermentation in 63 days. Figure 1 ), and the water content decreases rapidly ( Figure 2 The C / N ratio decreased significantly. Figure 3 Compost products had no significant effect on the germination of Shanghai bok choy seeds. Figure 4 Compared with Examples 2, 3, and 4, the temperature, moisture content, and C / N ratio of the compost pile in this example changed significantly with the number of composting days, indicating that the composting was more thorough.

[0038] Comparative Example 1

[0039] The steps in this comparative example are the same as in Example 2, except that step (2) is omitted.

[0040] The results showed that the temperature was still rising 70 days after composting in this example, and the compost products had a significant impact on the germination of Shanghai bok choy seeds, indicating that the composting was not yet complete, the degradation of citrus pruning branches was low, and the degree of decomposition was incomplete.

[0041] Comparative Example 2

[0042] The steps in this comparative example are the same as in Example 2, except that the compound microbial agent 1 is replaced with micro-element biological composting fermentation microbial agent, and everything else remains the same.

[0043] The results showed that this embodiment could complete composting fermentation in 70 days. Figure 1 ), and water content ( Figure 2 ), C / N value ( Figure 3 The changes were similar to those in Example 2, and the compost products had no significant effect on the germination of Shanghai bok choy seeds. Figure 4 Compared to Example 3, this example showed significantly slower changes in moisture content and C / N ratio, indicating that crushing citrus pruning branches can accelerate the formation of compost products.

[0044] The above description of the embodiments is only for the purpose of helping to understand the technical solutions and core ideas of this application; those skilled in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or make equivalent substitutions for some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of this application.

Claims

1. A compound microbial agent for composting citrus pruning branches, characterized in that, This composting compound microbial agent is composed of bacteria and fungi; wherein the bacteria include Delftobacterium (…). Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromo bacter ); The fungi include Alternaria ( Nimbya alternantherae ), Penicillium oxalate ( Penicillium oxalicum ), Candida ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger ( Aspergillus niger ).

2. The citrus pruning branch composting compound microbial agent as described in claim 1, characterized in that, The number of live bacteria in the composting compound microbial agent is not less than 8.3 lg cfu / mL.

3. The citrus pruning branch composting compound microbial agent as described in claim 2, characterized in that, The viable bacterial count in the composting compound microbial agent is not less than 4.4 lg CFU / mL, and the viable fungal count is not less than 3.9 lg CFU / mL.

4. The citrus pruning branch composting compound microbial agent as described in claim 3, characterized in that, The bacteria in the composting compound microbial agent include Delftobacterium ( Delftia tsuruhatensis The viable count of *Bacillus belyssus* was 2.0–3.0 lg cfu / mL. Bacillus velezensis The viable count of Bacillus subtilis was 2.5~3.5 lg CFU / mL. Bacillus subtilis The viable bacterial count was 2.5–3.0 lg CFU / mL and Achromobacterium ( Achromobacter The viable bacterial count was 1.0~1.5 lg cfu / mL; The fungi in the composting compound microbial agent include Alternaria ( Nimbya alternantherae The viable count of Penicillium oxalate was 0.5~1.21 g CFU / mL. Penicillium oxalicum The viable cell count was 1.0~1.5 lg CFU / mL, and the viable cell count of Candida albicans was 1.0~1.5 lg CFU / mL. Candida The viable count of ) was 0.75~1.5 lg cfu / mL, and that of specific saprophytic molds ( Vanrija Humicola The viable count of ) was 1.25~2.0 lg cfu / mL and Aspergillus niger ( Aspergillusniger The viable count was 0.5~1.5 lg cfu / mL.

5. A method for preparing a compound microbial agent for citrus pruning branch composting as described in any one of claims 1-4, characterized in that, Includes the following steps: S1, Bacterium delavayi ( Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromobacter Alternaria ( Nimbya alternantherae ), Penicillium oxalate ( Penicillium oxalicum ), Candida ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger ( Aspergillusniger The strains were isolated and purified by incubation at 37°C for 48 hours, and the purified strains were inoculated onto LB slant agar. S2, Bacterium deltaeoides ( Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromo bacter The strains were inoculated into 300 mL of LB liquid medium and cultured in a shaker at 37℃ and 180 r / min for 1-2 days until the viable count in the bacterial solution was not less than 4.4 lg cfu / mL. S3. Select Alternaria from the purified strains ( Nimbya alternantherae ), Penicillium oxalate ( Penicillium oxalicum ), Candida ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger ( Aspergillus niger The mycelia and spores of the mycelia were inoculated into 200 mL of PD medium and cultured at 30℃ and 200 r / min for 5 days to allow the mycelial balls to cover the entire PD liquid medium and the viable count to be no less than 3.9 lg cfu / mL. S4, Add Delft bacteria ( Delftia tsuruhatensis Bacillus belesiensis ( Bacillus velezensis Bacillus subtilis ( Bacillus subtilis ) and Achromobacterium ( Achromo bacter The culture medium of the bacteria was mixed in equal volume ratio to construct bacterial agent C1; S5, Alternaria ( Nimbya alternantherae ), Penicillium oxalate ( Penicillium oxalicum ), Candida ( Candida ), specific humic molds ( Vanrija Humicola ) and Aspergillus niger ( Aspergillus niger The culture medium of the bacteria was mixed in equal volume ratio to construct bacterial agent C2; S6. Mix the culture media of bacterial agent C1 and bacterial agent C2 in equal volume ratio to construct bacterial agent C3, thus obtaining a compound bacterial agent.

6. The method for preparing a compound microbial agent for citrus pruning branch compost as described in claim 5, characterized in that, The method for preparing the LB liquid culture medium is as follows: Weigh 10g of tryptone, 5g of yeast extract, and 10g of sodium chloride in sequence, and add them to a container containing 950ml of deionized water. Shake the container until the solute dissolves. Then add 5mol / L sodium hydroxide to the container to adjust the pH value to 7.4, and add deionized water to make up the volume to 1L. Then place the container in a pressure sterilizer and steam sterilize it at 15psi for 20min to obtain the LB liquid culture medium. The preparation method of the LB solid culture medium is as follows: Weigh 10g of tryptone, 5g of yeast extract, and 10g of sodium chloride in sequence, and add them to a container containing 950ml of deionized water. Shake the container until the solute dissolves. Then add 15g of agar powder and 5mol / L sodium hydroxide to the container to adjust the pH value to 7.

4. Add deionized water to the container to make up the volume to 1L. Then place the container in a pressure sterilizer and steam sterilize it at 15psi for 20min to obtain LB slant culture medium.

7. The method for preparing a compound microbial agent for citrus pruning branch compost as described in claim 5, characterized in that, The preparation method of the PD liquid culture medium is as follows: Weigh 200g of potatoes, wash, peel and slice them, put them in a pot, add 1000mL of water, boil for 15min, then filter with gauze, continue to add water to the pot to make the water in the pot reach 1000mL, then add 12g of glucose, and boil until the solid is fully dissolved, then dispense to obtain PD liquid culture medium; sterilize the dispensed PD culture medium within 1h by high pressure steam moist heat sterilization, at 0.1 MPa steam pressure, temperature up to 121 ℃, and maintain for 20min.

8. The application of a citrus pruning branch fermentation compound microbial agent as described in any one of claims 1-4 in citrus pruning branch composting, characterized in that, After the citrus pruned branches have been composted for 3-7 days, the compound microbial agent is diluted and sprayed evenly at a rate of 5 kg per ton of citrus pruned branches. The dilution ratio of the compound microbial agent is 800-1000 times.

9. The application of the citrus pruning branch fermentation compound microbial agent as described in claim 8 in citrus pruning branch composting, characterized in that, A compound microbial agent is applied to a mixture of citrus pruning branches and manure, wherein the compound microbial agent is applied at a ratio of 5 kg / ton of the total mass of the mixture.

10. The application of the citrus pruning branch fermentation compound microbial agent as described in claim 9 in citrus pruning branch composting, characterized in that, The compound microbial agent is applied to a mixture of shredded citrus pruning branches and manure, wherein the compound microbial agent is applied at a mass ratio of 3 kg / ton of the total mixture.

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