Microbial leavening agent and application thereof

Abstract

The invention belongs to the technical field of leavening agents, and discloses a microbial leavening agent and application thereof. The microbial fermentation agent comprises compound microorganisms and a carrier, the compound microorganisms comprise lactobacillus plantarum and Aksu marine bacillus in a volume ratio of (1-3): 1; the carrier is obtained by performing swelling treatment on muscovite powder carried with cetyltrimethylammonium bromide through CO2 supercritical fluid. According to the invention, lactobacillus plantarum and Marinobacter acksu in a proper proportion are used as active strains and are mixed with a specific carrier to prepare the microbial starter, and the lactobacillus plantarum and the Marinobacter acksu cooperate with each other, so that the humification degree of compost fermentation of sheep manure and the content of organic matters in the obtained decomposed organic fertilizer can be effectively improved.

Description

Technical Field

[0001] The present invention relates to the technical field of fermenting agents, and particularly to microbial fermenting agents and their applications. Background Art

[0002] In the technical field of fermenting agents, microbial fermenting agents, as a key means to promote the efficient fermentation of organic substances and improve the quality of fertilizers, have always received extensive attention from the scientific research community and the industrial community. Traditional microbial fermenting agents often contain only a single or a few microbial strains, and their fermentation efficiency and effects are relatively limited, making it difficult to meet the growing demand of modern agriculture for high-efficiency and environmentally friendly fertilizers.

[0003] Sheep manure, as a natural high-quality organic fertilizer raw material, is rich in cellulose substances. However, during the composting fermentation process, these cellulose substances are often difficult to be completely degraded, which affects the degree of humification of the fertilizer and the content of organic matter in the organic fertilizer, and further limits the full exertion of its fertilizer efficiency. Therefore, it is particularly urgent and important to explore a microbial fermenting agent that can efficiently degrade cellulose substances and significantly improve the composting fermentation efficiency of sheep manure. Summary of the Invention

[0004] To solve the above problems, the present invention provides a microbial fermenting agent and its application materials.

[0005] In the first aspect, the present invention provides a microbial fermenting agent, which includes composite microorganisms and a carrier; The composite microorganisms are composed of Lactobacillus plantarum and Marinobacter akesuensis; The carrier is obtained by swelling muscovite powder with cetyltrimethylammonium bromide carried by CO2 supercritical fluid.

[0006] Further, the volume ratio of the Lactobacillus plantarum bacterial suspension to the Marinobacter akesuensis bacterial suspension is (1 - 3):1, and the cell concentrations of the two bacterial suspensions are the same, for example, it can be 10 8 CFU.

[0007] Further, the volume-weight ratio of the composite microorganisms to the carrier is (1 - 2):(7 - 11).

[0008] Further, the mass ratio of muscovite powder to cetyltrimethylammonium bromide is (10 - 15):1.

[0009] Further, the preparation method of the carrier is: adding muscovite powder and cetyltrimethylammonium bromide into a closed reaction device, discharging air and then introducing CO2, and then performing swelling treatment for 90 - 120 minutes under the system conditions of a temperature of 35 - 45°C and a pressure of 20 - 25 Mpa, and then depressurizing, washing and drying.

[0010] In a second aspect, a method for preparing a microbial fermenting agent includes the following steps: Ferment Lactobacillus plantarum and Marinobacter akesuensis separately, and then mix the obtained Lactobacillus plantarum suspension and Marinobacter akesuensis suspension to obtain a composite microorganism; Mix the composite microorganism with a carrier, and then perform vacuum drying to obtain the microbial fermenting agent.

[0011] In a third aspect, the present invention provides an application of the microbial fermenting agent according to any one of the first aspect in the preparation of fertilizers.

[0012] In a fourth aspect, the present invention provides a microbial organic fertilizer, which includes a mature organic fertilizer obtained by composting and fermenting sheep manure using the microbial inoculant according to any one of the first aspect.

[0013] Furthermore, the microbial organic fertilizer further includes an extract of the residues of the Linggui Zhugan Decoction granules.

[0014] Furthermore, the mass ratio of the mature organic fertilizer to the extract of the residues of the Linggui Zhugan Decoction granules is (300-500):(1-3).

[0015] The above technical solutions provided by the present invention have at least the following advantages compared with the prior art: In the present invention, muscovite powder is used as a microbial carrier after being swollen by CO2 supercritical fluid carrying cetyltrimethylammonium bromide, which can enhance the stability of the inoculated microorganisms during the fermentation process, as well as their persistence, survival and colonization ability in the soil and plant roots.

[0016] Applying the microbial fermenting agent provided by the present invention to the preparation of fertilizers can increase the content of organic fertilizers in the fertilizers.

[0017] Using the microbial fermenting agent provided by the present invention to ferment sheep manure to prepare a microbial organic fertilizer can effectively improve the degree of humification of the sheep manure compost fermentation and the content of organic matter in the obtained mature organic fertilizer.

[0018] The present invention discovers that the extract of the residues of the Linggui Zhugan Decoction granules can prevent and control citrus scab. Adding it to the sheep manure fermentation fertilizer and applying it to citrus trees can increase the citrus yield. Detailed implementation manners

[0019] To make the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Apparently, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0020] Unless otherwise specified, all kinds of raw materials, reagents, instruments, and equipment used in the present invention can be obtained through market purchase or prepared by existing methods.

[0021] The first embodiment of the present invention provides a microbial ferment, and the microbial ferment includes a composite microorganism and a carrier; The composite microorganism is composed of Lactobacillus plantarum and Marinobacter akesuensis; The carrier is obtained by swelling mica powder with CO2 supercritical fluid carrying cetyltrimethylammonium bromide.

[0022] The embodiment of the present invention provides a microbial ferment. The invention prepares a microbial ferment by mixing Lactobacillus plantarum and Marinobacter akesuensis as active strains with a specific carrier. The two cooperate synergistically. The use of this carrier can reduce the dosage and types of microorganisms, and the obtained ferment can effectively improve the humification degree of sheep manure composting fermentation and the organic matter content in the obtained mature organic fertilizer. Specifically: Mica powder: As a natural mineral, mica powder has good adsorption and water retention properties, can provide attachment points and growth environments for microorganisms, and at the same time, the trace elements it contains can also promote the activity of microorganisms.

[0023] Swelling treatment with CO2 supercritical fluid carrying cetyltrimethylammonium bromide: CO2 supercritical fluid has excellent permeability and solubility, can carry cetyltrimethylammonium bromide deep into the interior of mica powder, cause it to swell, increase the specific surface area, improve the adsorption capacity and the microorganism attachment efficiency. This treatment not only improves the physical properties of the carrier, but also promotes the interaction between microorganisms and the carrier, can achieve improving the fermentation effect with fewer types and less quality of microorganisms, and enhances the stability and activity of microorganisms during the composting process.

[0024] Therefore, the microbial ferment provided by the present invention realizes the synergistic effect between microorganisms and the carrier, and between microorganisms through the specially treated carrier, ensures the stability of microorganisms during the fermentation process and field application, effectively optimizes the fermentation process of sheep manure composting, improves the humification degree and the organic matter content in the organic fertilizer, and provides high-quality organic fertilizers for agricultural production.

[0025] In the present invention, Lactobacillus plantarum can directly adopt commercially available products such as Lactobacillus plantarum CGMCC 1.557, Marinobacter akesuensis can directly adopt commercially available products such as Marinobacter akesuensis KCTC 12758, and mica powder can directly adopt commercially available mica powder with a particle size of 200 mesh.

[0026] In some preferred embodiments, to further improve the fermentation effect of the microbial fermenting agent, the dosages of Lactobacillus plantarum and Marinobacter akesuensis can also be controlled: the volume ratio of the Lactobacillus plantarum bacterial suspension to the Marinobacter akesuensis bacterial suspension is (1-3):1, and the cell concentrations of the two bacterial suspensions are the same. The higher proportion of Lactobacillus plantarum helps to rapidly reduce the pH, creating an environment conducive to the growth of Marinobacter akesuensis, while Marinobacter akesuensis can complement the deficiency of Lactobacillus plantarum in decomposing complex organic substances. The two work synergistically to improve the composting efficiency.

[0027] Regarding the dosage relationship between the composite microorganism and the carrier, there is no special limitation, and those skilled in the art can adjust it adaptively according to experience. For example, in some specific embodiments, the volume-weight ratio (v / w) of the composite microorganism to the carrier is (1-2):(7-11).

[0028] In some specific embodiments, the preparation method of the carrier includes the following steps: Add muscovite powder and cetyltrimethylammonium bromide into a closed reaction device according to a weight ratio of (10-15):1, discharge the air, then introduce CO2, and then carry out a swelling treatment for 90-120 minutes under the system conditions of a temperature of 35-45 °C and a pressure of 20-25 Mpa. Release the pressure, wash with ethanol and dry to obtain the carrier.

[0029] The second embodiment of the present invention provides a preparation method of a microbial fermenting agent, including the following steps: Ferment Lactobacillus plantarum and Marinobacter akesuensis separately, and then mix the obtained Lactobacillus plantarum bacterial suspension and Marinobacter akesuensis bacterial suspension to obtain a composite microorganism; Mix the composite microorganism and the carrier, and then vacuum dry to obtain the microbial fermenting agent.

[0030] In some specific embodiments, fermenting Lactobacillus plantarum and Marinobacter akesuensis separately, and then mixing the obtained Lactobacillus plantarum bacterial suspension and Marinobacter akesuensis bacterial suspension according to a volume ratio to obtain a composite microorganism can be carried out according to the conventional operation methods in the art. As an example, the following operation method is adopted in the subsequent examples and comparative examples of the present invention: 1. Strain activation Lactobacillus plantarum: Take Lactobacillus plantarum from the cryopreservation tube, pick a small amount of bacterial liquid with an inoculation loop, and inoculate it into MRS liquid medium. The formula of MRS liquid medium is: peptone 10 g, beef extract 10 g, yeast extract 5 g, glucose 20 g, Tween 80 1 mL, dipotassium hydrogen phosphate 2 g, sodium acetate 5 g, ammonium citrate 2 g, magnesium sulfate 0.58 g, manganese sulfate 0.25 g, made up to 1 L with distilled water, and adjust the pH to 6.2-6.4. Culture in a constant temperature incubator at 37 °C for 18-24 h for activation.

[0031] Marinobacter akesuensis: Take Marinobacter akesuensis and inoculate it into 2216E liquid medium. The formula of 2216E liquid medium is: peptone 5 g, yeast extract 1 g, ferric phosphate 0.01 g, sodium chloride 19.45 g, magnesium sulfate 5.9 g, sodium dihydrogen phosphate 0.08 g, calcium chloride 1.8 g, potassium chloride 0.55 g, sodium bicarbonate 0.16 g, potassium bromide 0.08 g, boric acid 0.026 g, sodium silicate 0.004 g, sodium fluoride 0.0024 g, strontium chloride 0.034 g. Make up the volume to 1 L with distilled water and adjust the pH to 7.6 - 7.8. Incubate in a constant temperature shaker at 28 °C at a rotation speed of 150 r / min for 24 - 36 h for activation.

[0032] 2. Fermentation culture Lactobacillus plantarum: Inoculate the activated Lactobacillus plantarum into fresh MRS liquid medium at an inoculation amount of 3%, and the liquid loading volume is 25% of the volume of the Erlenmeyer flask. Incubate statically in a constant temperature incubator at 37 °C. Monitor the growth of the bacterial liquid by taking samples at regular intervals and using the turbidimetric method (OD600). The late logarithmic phase is reached after about 14 h of culture.

[0033] Marinobacter akesuensis: Inoculate the activated Marinobacter akesuensis into fresh 2216E liquid medium at an inoculation amount of 4%, and the liquid loading volume is 25% of the volume of the Erlenmeyer flask. Incubate in a constant temperature shaker at 28 °C at a rotation speed of 200 r / min. Similarly, monitor the growth of the bacterial liquid by taking samples at regular intervals and using the turbidimetric method (OD600). The late logarithmic phase is reached after about 26 h of culture; 3. Bacterial cell collection When Lactobacillus plantarum and Marinobacter akesuensis reach the late logarithmic phase respectively, transfer the fermentation broth to a centrifuge tube and centrifuge at 4 °C and 9000 r / min for 10 min to collect the wet bacterial cells.

[0034] 4. Resuspension of bacterial cells Resuspend the collected wet bacterial cells with 0.85% physiological saline to prepare a bacterial suspension.

[0035] 5. Mixing of composite bacterial cells Mix the Lactobacillus plantarum bacterial suspension and the Marinobacter akesuensis bacterial suspension evenly to obtain a composite microorganism.

[0036] The third embodiment of the present invention provides the application of the microbial fermenting agent described in any one of the first embodiments in the preparation of fertilizers.

[0037] The fourth embodiment of the present invention provides a microbial organic fertilizer, including a mature organic fertilizer obtained by composting sheep manure by using the microbial fermenting agent described in any one of the first embodiments.

[0038] As a preferred embodiment, the aforementioned microbial organic fertilizer may further include the extract of the medicinal residues of Linggui Zhugan Decoction granules.

[0039] As a preferred embodiment, the mass ratio of the matured organic fertilizer to the extract of the medicinal residues of Linggui Zhugan Decoction granules is (300 - 500):(1 - 3).

[0040] The present invention discovers that the medicinal residues of Linggui Zhugan Decoction granules have the use for preventing and controlling citrus scab, and further applies it to the matured organic fertilizer to prepare a microbial organic fertilizer capable of increasing citrus yield, which has wide practical uses.

[0041] For the extraction method of the extract of the medicinal residues of Linggui Zhugan Decoction granules, the commonly used effective component extraction methods in the art can be adopted, and the parameters in the extraction process can be adjusted adaptively according to the extraction object. As an example, reduced pressure reflux extraction can be used, and the specific parameters are as follows: the pressure is 20 - 30 kPa, the temperature is 65 - 75 °C, the time is 60 - 90 minutes, and the number of times is 1 - 3 times.

[0042] The above-mentioned microbial organic fertilizer containing the extract of the medicinal residues of Linggui Zhugan Decoction granules has at least any one of the following uses: 1) Preventing and controlling citrus scab; 2) Improving the quality of citrus.

[0043] It should be noted that for the component raw materials involved in the microbial fermenting agent, its preparation method and application provided in the embodiments of the present invention, if there is no special limitation or specific description, commercially available products can be directly adopted or self-made by using the existing publicly disclosed preparation methods; meanwhile, for the steps and parameters involved, if there is no special limitation or specific description, they can be carried out according to the processing techniques disclosed in the prior art or directly adopted by using the existing equipment, and the present invention document will not elaborate one by one.

[0044] The following combines specific embodiments to further illustrate the present invention. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are usually determined according to national standards. If there is no corresponding national standard, they are carried out according to general international standards, conventional conditions, or the conditions recommended by the manufacturer.

[0045] The Lactobacillus plantarum (product number HZB127512) and Marinobacter akesuensis (product number HZB137372) used in the following embodiments are both purchased from Huizao Biology.

[0046] Example 1 Provide a microbial fermenting agent, the microbial fermenting agent includes composite microorganisms and a carrier; The volume - weight ratio (v / w) of the composite microorganisms to the carrier is 1.5:9; The composite microorganism is a suspension of Lactobacillus plantarum and Bacillus aquimaris with a volume ratio of 2.6:1; The carrier is obtained by swelling mica powder with cetyltrimethylammonium bromide carried by CO2 supercritical fluid. The specific process is as follows: Mica powder and cetyltrimethylammonium bromide are added to a closed reaction device in a weight ratio of 13:1. After exhausting the air, CO2 is introduced, and then swelling treatment is carried out for 100 minutes under the system conditions of a temperature of 40 °C and a pressure of 22 Mpa. After depressurization, ethanol washing and drying are carried out to obtain the carrier.

[0047] The preparation method of the above microbial ferment includes the following steps: Ferment Lactobacillus plantarum and Bacillus aquimaris separately, and then mix the obtained Lactobacillus plantarum suspension and Bacillus aquimaris suspension according to the volume ratio to obtain a composite microorganism; Mix the composite microorganism and the carrier, and then vacuum dry to obtain the microbial ferment.

[0048] Example 2 Provide a microbial ferment, which includes a composite microorganism and a carrier; The volume-weight ratio (v / w) of the composite microorganism to the carrier is 1:7; The composite microorganism is Lactobacillus plantarum and Bacillus aquimaris with a volume ratio of 2:1; The carrier is obtained by swelling mica powder with cetyltrimethylammonium bromide carried by CO2 supercritical fluid. The specific process is as follows: Mica powder and cetyltrimethylammonium bromide are added to a closed reaction device in a weight ratio of 10:1. After exhausting the air, CO2 is introduced, and then swelling treatment is carried out for 120 minutes under the system conditions of a temperature of 35 °C and a pressure of 20 Mpa. After depressurization, ethanol washing and drying are carried out to obtain the carrier.

[0049] The preparation method of the above microbial ferment includes the following steps: Ferment Lactobacillus plantarum and Bacillus aquimaris separately, and then mix the obtained Lactobacillus plantarum suspension and Bacillus aquimaris suspension according to the volume ratio to obtain a composite microorganism; Mix the composite microorganism and the carrier, and then vacuum dry to obtain the microbial ferment.

[0050] Example 3 Provide a microbial ferment, which includes a composite microorganism and a carrier; The volume-weight ratio (v / w) of the composite microorganism to the carrier is 2:11; The composite microorganism is Lactobacillus plantarum and Marinobacter akesuensis with a volume ratio of 3:1; The carrier is obtained by swelling mica powder with cetyltrimethylammonium bromide carried by CO2 supercritical fluid, and the specific process is as follows: Mica powder and cetyltrimethylammonium bromide are added to a closed reaction device according to a weight ratio of 15:1, air is discharged, and then CO2 is introduced. Then, swelling treatment is carried out for 90 minutes under the system conditions of a temperature of 45 °C and a pressure of 25 Mpa, the pressure is released, washed with ethanol and dried to obtain the carrier.

[0051] The preparation method of the above microbial ferment agent includes the following steps: Lactobacillus plantarum and Marinobacter akesuensis are fermented separately, and then the obtained Lactobacillus plantarum suspension and Marinobacter akesuensis suspension are mixed according to a volume ratio to obtain a composite microorganism; The composite microorganism and the carrier are mixed, and then vacuum dried to obtain the microbial ferment agent.

[0052] Example 4 The difference from Example 1 is only that: the volume-weight ratio (v / w) of the composite microorganism and the carrier is 1:5; the remaining steps and parameters are the same.

[0053] Example 5 The difference from Example 1 is only that: the volume-weight ratio (v / w) of the composite microorganism and the carrier is 2:15; the remaining steps and parameters are the same.

[0054] Example 6 The difference from Example 1 is only that: the volume ratio of the Lactobacillus plantarum suspension and the Marinobacter akesuensis suspension in the composite microorganism is 1:1; the remaining steps and parameters are the same.

[0055] Example 7 Provide a microbial organic fertilizer, which is composed of component A and component B with a weight ratio of 400:2.3; Component A is a decomposed organic fertilizer, which is obtained by composting and fermenting the microbial ferment agent obtained in Example 1, and the specific method refers to Test Example 1; Component B is an extract of the medicinal residue of Linggui Zhugan Decoction. The preparation method is: the medicinal residue of Linggui Zhugan Decoction and water are mixed according to a weight ratio of 1:10, and then decompression reflux is carried out for 75 minutes under the conditions of a pressure of 25 kPa and a temperature of 70 °C, extracted 2 times, and then the obtained water extract is combined and concentrated and dried under reduced pressure. Obtain the extract of the traditional Chinese medicine medicinal residue.

[0056] Comparative Example 1 The only difference from Example 1 is that the carrier is the muscovite powder in Example 1 that has not been swelled with hexadecyltrimethylammonium bromide carried by CO2 supercritical fluid; the remaining steps and parameters are the same.

[0057] Comparative Example 2 The only difference from Example 1 is that Aksu Marine Bacillus is replaced by an equal amount of Bacillus subtilis; the remaining steps and parameters are the same.

[0058] Comparative Example 3 The only difference from Example 1 is that the volume-to-weight ratio (v / w) of the composite microorganism and the carrier is changed to 1:3; the remaining steps and parameters are the same.

[0059] Comparative Example 4 The only difference from Example 1 is that the volume-to-weight ratio (v / w) of the composite microorganism and the carrier is changed to 1:12; the remaining steps and parameters are the same.

[0060] Comparative Example 5 The only difference from Example 1 is that the volume ratio of Lactobacillus plantarum to Aksu marine bacillus in the composite microorganism is changed to 4:1; the other steps and parameters are the same.

[0061] Test Example 1 In this example, the microbial fermentation agents obtained in the above Examples 1-3 and Comparative Examples 1-2 were applied to sheep manure composting fermentation to examine its fermentation effect.

[0062] Composting fermentation and testing specifically include the following processes: 1. Ratio of raw materials and leavening agent: Compost raw materials: Fresh sheep manure is used as the basic material for composting.

[0063] Amount of fermentation agent added: Add microbial fermentation agent at a ratio of 0.3% of the mass of compost raw materials.

[0064] Mixing and adjustment: Mix the fermentation agent and sheep manure thoroughly and evenly, then adjust the moisture content of the material to (55±5)%. The judgment standard is that the material can be held in a ball by hand, no liquid seeps out, and the material remains intact after releasing the hand.

[0065] 2. Heap construction and management: Pile shape and size: The pile is constructed in a cone shape with a height controlled between 1.2 and 1.4 meters and a diameter ranging from 2 to 3 meters.

[0066] Fermentation cycle: Set the composting cycle to 25 days.

[0067] Turning and ventilation: Use mechanical methods to turn the compost and rely on natural ventilation.

[0068] Temperature Management: When the heap temperature reaches 50 °C, the first turning of the heap is carried out.

[0069] If the temperature of the heap continues to rise to about 60 °C, the second turning of the heap is carried out.

[0070] In the high-temperature stage (50 - 65 °C), maintain this temperature range for about 10 days. During this period, if the temperature approaches or exceeds 70 °C, immediate turning of the heap is required to cool down and increase the oxygen supply.

[0071] Entering the post-maturation stage, turn the heap once every 3 - 4 days until the temperature of the heap gradually decreases to approach the ambient temperature.

[0072] 3. Detection method 1) Determination of organic matter content: Sampling: After the composting is completed, collect mixed samples from multiple points.

[0073] Determination method: Use the K2Cr2O7 volumetric method to determine the organic matter content (dry basis, %) in each mixed sample.

[0074] 2) Evaluation of maturity: Preparation of leaching solution: Extract the fresh compost sample with deionized water at a solid-liquid ratio of 1:10 (mass / volume) for 3 hours, and then filter to obtain the compost leaching solution.

[0075] Absorbance determination: Use an ultraviolet spectrophotometer to measure the absorbance values (denoted as E4 and E6) of the leaching solution at wavelengths of 465 nm and 665 nm respectively.

[0076] Calculation of humification parameters: Calculate the E4 / E6 value as one of the indicators for evaluating the maturity of the compost. The smaller the E4 / E6 value, the higher the humification degree of the compost.

[0077] The test results are shown in Table 1.

[0078] Table 1 .

[0079] As can be seen from Table 1, compared with Comparative Examples 1 - 2, the organic matter content and the humification degree of the compost in the sheep manure matured organic fertilizer obtained by using the microbial fermentation agents provided in Examples 1 - 3 of the present invention are both higher; and the organic matter content in the sheep manure matured organic fertilizer obtained by using the microbial fermentation agent provided in Example 1 can reach 73.1%, and the composting fermentation effect is the best.

[0080] Test Example 2 This example aims to evaluate the control effect of the extract of the Atractylodes macrocephala Koidz. and Cinnamon Twig Decoction granules in Example 7 on citrus scab, and use dithianon as a control.

[0081] Test method: Operate according to standard agricultural practices. Spray the citrus tree canopy (including fruits) with the extract of the medicinal residues of Linggui Zhugan Decoction granules or dithianon once every 14 days, with a spraying volume of 3000 L / Ha, ensuring that the liquid medicine evenly covers the canopy. Set three replicates for each treatment; in accordance with GB / T 17980.102—2004, conduct a graded investigation on the incidence of citrus scab. 20 days after the last application, investigate and record the incidence of citrus scab in each treatment area. Investigate 2 fruit trees in each treatment plot, take 5-point samples from each fruit tree in the east, west, south, north, and middle directions, investigate all the leaves of 2 shoots and 5 fruits at each point, and calculate the disease index and control effect. The disease grading standard for citrus scab is shown in Table 2. The control effect is expressed as control efficacy (%), and the test results are shown in Table 3.

[0082] Table 2 Disease grading standard for citrus scab 。

[0083] Disease index = [∑(number of diseased leaves (fruits) at each level × relative level value)] / [total number of investigated leaves (fruits) × 9] × 100%; Disease index control efficacy (%) = (disease index in the blank control area - disease index in the treatment area) / disease index in the blank control area × 100%.

[0084] Table 3 。

[0085] As can be seen from Table 3, compared with the control group with dithianon as the test sample, the extract of the medicinal residues of Linggui Zhugan Decoction granules has excellent control effect on citrus scab.

[0086] Based on the above tests, the microbial organic fertilizer obtained in Example 7 was further investigated for its application in citrus cultivation. It was used as the experimental group and compared with a control group that only used a commercially available special fertilizer for citrus cultivation (rotted organic fertilizer) as the base fertilizer. Test method: The variety was the citrus navel orange variety. The microbial organic fertilizer was used as the base fertilizer, and at the same time, during the young fruit formation period, the extract of the Atractylodes macrocephala, Cinnamon Twig, Poria, and Licorice Decoction granule residues in Example 7 was sprayed on the leaves and the tree body. The spraying amount was 3000 L / Ha, and the concentration of the Atractylodes macrocephala, Cinnamon Twig, Poria, and Licorice Decoction granule residues extract was 80 g / L. When the new shoots germinated to a bud length of 1 - 2 mm, it was sprayed for the first time to protect the young leaves. When 2 / 3 of the flowers withered, it was sprayed for the second time to focus on protecting the young fruits. It was sprayed for the third time during the fruit setting period to ensure that the liquid medicine evenly covered the tree crown. Each treatment was set with three replicates, and the rest were carried out according to the existing citrus cultivation; at the same time, 2 control groups were set: Control Group 1, the difference from the experimental group was only that the base fertilizer was replaced with only the commercially available special fertilizer for citrus cultivation (Shijiazhuang WoFuWo Fertilizer Industry Co., Ltd., rotted organic fertilizer); the rest of the steps and parameters were the same; Control Group 2, the difference from the experimental group was only that the Atractylodes macrocephala, Cinnamon Twig, Poria, and Licorice Decoction granule residues extract was not sprayed on the citrus fruit trees. After the fruits matured, the citrus yield per mu (50 citrus trees were planted per mu, with a plant spacing of 3 × 3.5) and the Vc content in the fruits were detected, and the test results of the statistical data are shown in Table 4.

[0087] Table 4 。

[0088] As can be seen from Table 4, compared with the commercially available special fertilizer for citrus cultivation, the microbial organic fertilizer provided in the embodiment of the present invention can increase the Vc content in the obtained citrus, thereby improving the quality of the obtained fruits.

[0089] In summary, the embodiment of the present invention provides a microbial fermenter and its application, and a microbial organic fertilizer. On the one hand, the present invention mixes Lactobacillus plantarum and Marinobacter akesuensis in an appropriate ratio as active strains and a specific carrier to obtain a microbial fermenter. The two cooperate synergistically, and can effectively improve the humification degree of sheep manure compost fermentation and the organic matter content in the obtained rotted organic fertilizer; on the other hand, the present invention discovers that the residues of the Atractylodes macrocephala, Cinnamon Twig, Poria, and Licorice Decoction granules have the use of preventing and treating citrus scab, and further applies them to the obtained rotted organic fertilizer to obtain a microbial organic fertilizer that can increase citrus yield, which has a wide range of practical uses.

[0090] The various embodiments of the present invention may exist in the form of a range; it should be understood that the description in the form of a range is only for convenience and brevity and should not be construed as a rigid limitation on the scope of the present invention; therefore, it should be considered that the range description has specifically disclosed all possible sub-ranges and single values within that range. For example, it should be considered that the range description from 1 to 6 has specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and single numbers within the range, such as 1, 2, 3, 4, 5, and 6, and this applies regardless of the range. Additionally, whenever a numerical range is indicated herein, it means including any cited number (fraction or integer) within the indicated range.

[0091] The above are only specific embodiments of the present invention, enabling those skilled in the art to understand or implement the present invention. Various modifications to these embodiments will be obvious to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to these embodiments shown herein, but rather to the broadest scope consistent with the principles and novel features claimed herein.

Claims

1. A microbial fermentation agent, characterized in that, The microbial inoculant includes compound microorganisms and a carrier; The compound microorganisms are composed of Lactobacillus plantarum and Marinobacter akesuensis; The carrier is obtained by swelling mica powder with cetyltrimethylammonium bromide carried by CO2 supercritical fluid.

2. The microbial fermenting agent according to claim 1, wherein The volume ratio of the Lactobacillus plantarum suspension to the Marinobacter akesuensis suspension is (1-3):1, and the cell concentrations of the two suspensions are the same.

3. The microbial fermenting agent according to claim 1, wherein, The volume-weight ratio of the compound microorganisms to the carrier is (1-2):(7-11).

4. The microbial fermenting agent according to claim 1, wherein The mass ratio of mica powder to cetyltrimethylammonium bromide is (10-15):

1.

5. The microbial fermenting agent according to claim 1 or 4, characterized in that, The preparation method of the carrier is as follows: Add mica powder and cetyltrimethylammonium bromide into a closed reaction device, discharge the air and then introduce CO2, and then carry out swelling treatment for 90-120 minutes under the system conditions of a temperature of 35-45°C and a pressure of 20-25 Mpa, relieve the pressure, wash and dry.

6. The preparation method of the microbial fermenting agent according to any one of claims 1 to 5, characterized in that, It includes the following steps: Ferment Lactobacillus plantarum and Marinobacter akesuensis separately, and then mix the obtained Lactobacillus plantarum suspension and Marinobacter akesuensis suspension to obtain compound microorganisms; Mix the compound microorganisms and the carrier, and then dry them under vacuum to obtain the microbial inoculant.

7. The application of the microbial inoculant according to any one of claims 1-5 in the preparation of fertilizers.

8. Microbial organic fertilizer, characterized in that, It includes the mature organic fertilizer obtained by composting and fermenting sheep manure with the microbial inoculant according to any one of claims 1-5.

9. The microbial organic fertilizer according to claim 8, wherein, It also includes the extract of the medicinal residues of Lingguizhugan Decoction granules.

10. The microbial organic fertilizer according to claim 9, characterized in that, The mass ratio of the mature organic fertilizer to the extract of the medicinal residues of Lingguizhugan Decoction granules is (300-500):(1-3).