CN111518700B - Carbon-based composite microbial inoculant and application thereof - Google Patents

Carbon-based composite microbial inoculant and application thereof Download PDF

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CN111518700B
CN111518700B CN202010175633.1A CN202010175633A CN111518700B CN 111518700 B CN111518700 B CN 111518700B CN 202010175633 A CN202010175633 A CN 202010175633A CN 111518700 B CN111518700 B CN 111518700B
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soil
carbon
based composite
composite microbial
fermentation product
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CN111518700A (en
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周德平
吴淑杭
褚长彬
赵峥
王庆峰
蒲加军
刘善良
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Seek Bio Technology Shanghai Co ltd
Shanghai Academy of Agricultural Sciences
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Seek Bio Technology Shanghai Co ltd
Shanghai Academy of Agricultural Sciences
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
    • C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor

Abstract

The invention relates to a carbon-based composite microbial inoculum and application thereof, wherein the carbon-based composite microbial inoculum comprises an aspergillus oryzae solid fermentation product, a zygosaccharomyces rouxii liquid fermentation product, a bacillus belgii liquid fermentation product and biomass charcoal. The carbon-based composite microbial inoculum provided by the invention creatively compounds three fermentation products and biomass charcoal, and several components are synergistic, so that when the carbon-based composite microbial inoculum is used in a soil remediation agent, the pH value of soil can be adjusted to be neutral under the condition of no pesticide introduction, the soil salt content is reduced, the organic matter content is increased, and the incidence rate of the clubroot of the continuous cropping soil green vegetables is reduced. The quantity of plasmodiophora brassicae in continuous cropping soil treated by the soil remediation agent is obviously reduced, the quantity of plasmodiophora brassicae in the root system of the green vegetables in the harvesting period is obviously reduced, and the diversity of microbial community species in the soil and the plant root system is obviously increased.

Description

Carbon-based composite microbial inoculant and application thereof
Technical Field
The invention belongs to the technical field of agricultural biology, and relates to a carbon-based composite microbial inoculant and application thereof, in particular to a carbon-based composite microbial inoculant for a soil remediation agent and application thereof.
Background
Clubroot, also known as root cancer, is the most serious worldwide soil-borne disease which damages cruciferous crops at present, is caused by infection of plasmodiophora brassicae, has a wide host range, and can damage more than 100 cultivated and wild cruciferous plants such as Chinese cabbage, radish, shepherd's purse, cauliflower, rape and the like. After the infection of germs, the parenchyma cells of the root are proliferated to form swollen roots which mostly occur on main roots and lateral roots, the surface is smooth in the initial stage, and the surface is cracked and rough in the later stage, so that the moisture and nutrition absorption of the root system is influenced. The symptoms of the overground part at the early stage of the disease are not obvious, the leaves are wilted at noon when the disease is developed to a certain degree, the plants die in severe cases, the average yield loss reaches 20-30 percent, and the serious yield loss reaches more than 60 percent or even is lost. The plasmodiophora brassicae has a strict obligate parasitic range, and dormant spores of the plasmodiophora brassicae can still germinate in soil after 15-20 years and have pathogenic capability. Therefore, once they develop, there is a risk of replanting the soil with subsequent cruciferous crops. The land replacement and planting cost is high, and the potential safety hazard of chemical control food is more.
CN101058729B discloses a soil conditioner for preventing and treating clubroot of cruciferous crops. The conditioner comprises the following raw materials in parts by weight: 10-50 parts of organic matter, 10-60 parts of calcium magnesium phosphate fertilizer, 15-80 parts of carbonate, 0.8-5 parts of potassium, 0.8-5 parts of magnesium, 0.8-5 parts of zinc and 0.8-5 parts of boron; the organic matter is prepared from wild or cultivated marigold plants as raw materials, the wild plants are harvested in the full-bloom stage, the cultivated plants are harvested after flowers are harvested, and the organic matter is dried in the sun and crushed into powder. Through extensive tests on rape, Chinese cabbage, tuber mustard, broccoli, red sedge, kohlrabi, cabbage, isatis root and mustard, the control effect reaches 80% -95%. Has the advantages of economy, safety, high efficiency and simple and convenient use. But the organic materials used by the method are special and have no universality and convenience in application.
CN101416641A discloses a biological agent for preventing and treating clubroot of cruciferae and application thereof. The biological agent is prepared by using a test tube and shaking culture of a bacillus amyloliquefaciens XF-1 strain and a fermentation culture solution, and then is applied to rhizosphere soil of cruciferous crops, so that the biological agent has good prevention and control and yield increase effects, and is easy for factory production. But the single strain preparation has poor geographical adaptability and the efficacy is not easy to be stable.
CN107987838A discloses an organic preparation for preventing and controlling clubroot of cruciferous vegetables and application thereof. The organic preparation consists of organic raw materials and ingredients for promoting decomposition, wherein the organic materials comprise but are not limited to one or more of solid organic materials such as crop straws and wood chips, and liquid organic materials such as molasses and alcohol fermentation liquor; the decomposition promoting ingredient comprises one or more of but not limited to special microorganisms and organic catalytic substances; the mixture ratio of the organic raw materials to the decomposition promoting materials is (95-100) to (5-0). The organic preparation is ecological and environment-friendly, is suitable for soil with high clubroot after continuous cropping of various cruciferous vegetables, can effectively reduce the number of plasmodiophora brassicae in the soil and the incidence rate of crop clubroot after treatment, and has the sterilization rate and the clubroot prevention and control rate of 60-90 percent respectively. The technology focuses on early soil treatment, but does not give consideration to biological prevention and control of later crop pathogens.
In the prior art, strategies for effectively preventing clubroot or repairing the soil with clubroot are very limited, so that the development of a green pesticide-free clubroot continuous cropping soil repairing agent and a repairing method which have the advantages of locally available raw materials for treatment, no regional limitation on treatment effect and antagonistic inhibition effect on pathogenic bacteria of subsequent planted crops has important application value.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a carbon-based composite microbial inoculum and application thereof, and particularly provides a carbon-based composite microbial inoculum for a soil remediation agent and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
in one aspect, the invention provides a carbon-based composite microbial inoculum, which comprises an aspergillus oryzae solid fermentation product, a zygosaccharomyces rouxii liquid fermentation product, a bacillus belgii liquid fermentation product and biomass charcoal.
The carbon-based composite microbial inoculum related by the invention creatively compounds three fermentation products and biomass charcoal, and can quickly start an aerobic and anaerobic treatment process in a field after the three fermentation products are mixed with organic materials for flooding. Wherein, the aspergillus oryzae solid fermentation product and the bacillus beiLeisi liquid fermentation product can quickly start the aerobic decomposition of organic materials in soil, and influence the temperature and oxidation-reduction potential conditions in the treatment process; the zygosaccharomyces rouxii liquid fermentation product participates in anaerobic fermentation; the Bacillus belgii liquid fermentation product also has antagonism to various soil-borne and air-borne plant pathogenic bacteria, and can enhance disease resistance of later-stage crops. The three functional strains can decompose and utilize organic materials, and simultaneously build a lower oxidation-reduction potential in a soil microenvironment, and have a strong killing effect on soil-borne pathogenic bacteria; in addition, after the organic materials are decomposed, the organic matter content of soil is increased, the nutrients available for crops and small molecular acid and oligosaccharide substances capable of stimulating the growth of the crops are increased, and the growth of the crops can be promoted; a series of biochemical reactions are generated in the decomposition process of the organic materials, so that the pH value and the salt content of the soil are adjusted to a certain extent, and the growth of crops is facilitated; the Bacillus belgii has strong antagonistic and inhibitory effects on various plant pathogenic bacteria, can reduce the infection and ecological occupation of the pathogenic bacteria on replanted crops, and can enhance the immunity of the crops. The three components have synergistic effect, and when the compound is used in a soil remediation agent, the incidence rate of the clubroot of the continuous cropping vegetables can be reduced from 60% to about 10% under the condition of no introduction of pesticide.
Preferably, the total viable count of aspergillus oryzae, zygosaccharomyces rouxii and bacillus belgii in the carbon-based composite microbial inoculum is 1.0 multiplied by 109-3.0×1010E.g. 1.0X 109、3.0×109、5.0×109、8.0×109、1.0×1010Or 3.0X 1010And the other specific point values in the range can be selected, and are not described in detail herein.
The total live bacteria of aspergillus oryzae, zygosaccharomyces rouxii and bacillus belgii in the carbon-based composite microbial agentNumber specific selection of 1.0 × 109-3.0×1010The range is that if the total viable count is too high, the production cost is increased, and if the total viable count is too low, the soil treatment time needs to be prolonged, the soil treatment effect is affected, and even negative effects are generated on the subsequent crop growth.
Preferably, the ratio of the number of viable bacteria of aspergillus oryzae, zygosaccharomyces rouxii and bacillus belgii is 1 (1-2) to (1-2), for example, 1:1:1, 1:1:2, 1:2:1 or 1:2:2, and the like, and other specific values in the range can be selected, and are not described in detail herein.
When the ratio of the number of the live bacteria of the aspergillus oryzae, the zygosaccharomyces rouxii and the bacillus belezii is 1 (1-2) to 1-2, the synergistic interaction of the aspergillus oryzae, the zygosaccharomyces rouxii and the bacillus belezii can be maximized, namely, the pH of soil is more obviously adjusted, the content of organic matters is increased, the nutrient supply condition is improved, the physical structure of the soil is improved, the incidence rate of clubroot diseases of crops is more obviously reduced, the number of clubroot bacteria in the soil and the root systems of the crops is more obviously reduced, and the diversity of microbial community species in the soil and the root systems of the crops is more obviously.
Preferably, the mass percentage of the biomass charcoal in the charcoal-based composite microbial inoculum is 60-80%, for example, 60%, 65%, 70%, 75% or 80%, and other specific values within the range can be selected, and are not repeated herein.
Preferably, the fermentation medium formula of the aspergillus oryzae solid leavening comprises the following components in percentage by weight: 2-8% of bran, 2-8% of rice hull powder, 25-35% of rice straw, 2-6% of soybean flour and 50-70% of water.
The weight percentage of the bran can be 2%, 3%, 5%, 6% or 8%, and other specific values in the range can be selected, and are not repeated herein.
The weight percentage of the rice hull powder can be 2%, 3%, 5%, 6% or 8%, and other specific values in the range can be selected, and are not repeated herein.
The weight percentage of the rice straw can be 25%, 28%, 30%, 32% or 35%, and other specific values in the range can be selected, and are not repeated herein.
The weight percentage of the soybean meal can be 2%, 3%, 4%, 5% or 6%, and other specific values in the range can be selected, and are not repeated herein.
The weight percentage of the water can be 50%, 55%, 60%, 65% or 70%, and other specific values in the range can be selected, and are not repeated herein.
Preferably, the initial fermentation temperature of the aspergillus oryzae solid fermentation product is 20-40 ℃, such as 20 ℃, 25 ℃, 30 ℃, 35 ℃ or 40 ℃, and other specific values in the range can be selected, and are not repeated herein.
Specifically, the fermentation conditions of the aspergillus oryzae solid fermentation product are as follows: standing at 20-40 deg.C for 40min, turning over materials for 5min, and continuing for 2 weeks until the materials are completely filament-wrapped and the water content is about 30-40%.
Preferably, the fermentation medium formula of the zygosaccharomyces rouxii liquid fermentation product comprises the following components in percentage by weight: 1-5% of yeast extract, 1-3% of peptone, 1-3% of glucose and 90-97% of water.
The weight percentage of the yeast extract can be 1%, 2%, 3%, 4% or 5%, and other specific values in the range can be selected, and are not repeated herein.
The weight percentage of the peptone can be 1%, 1.5%, 2%, 2.5% or 3%, and other specific values in the range can be selected, and the detailed description is omitted here.
The weight percentage of the glucose can be 1%, 1.5%, 2%, 2.5% or 3%, and other specific values in the range can be selected, and are not repeated herein.
The weight percentage of the water can be 90%, 92%, 94%, 95% or 97%, and other specific values in the range can be selected, and are not repeated herein.
Preferably, the fermentation temperature of the zygosaccharomyces rouxii liquid fermentation product is 25-40 ℃, such as 25 ℃, 28 ℃, 30 ℃, 35 ℃ or 40 ℃, and other specific values in the range can be selected, and are not repeated herein.
Specifically, the fermentation conditions of the zygosaccharomyces rouxii liquid fermentation product are as follows: aerobic at 25-40 deg.C until thallus content is 1011The fermentation was terminated at cfu/mL.
Preferably, the fermentation medium formula of the liquid fermentation product of the bacillus beiLeisi comprises the following components in percentage by weight: peptone 1-3%, beef extract 0.5-2.5%, sodium chloride 0.5-2.5% and water 93-98%.
The weight percentage of the peptone can be 1%, 1.5%, 2%, 2.5% or 3%, and other specific values in the range can be selected, and the detailed description is omitted here.
The weight percentage of the beef extract can be 0.5%, 1.0%, 1.5%, 2.0% or 2.5%, and other specific values in the range can be selected, and are not repeated herein.
The weight percentage of the sodium chloride can be 0.5%, 1.0%, 1.5%, 2.0%, 2.5% or the like, and other specific values in the range can be selected, which is not repeated herein.
The weight percentage of the water can be 93%, 94%, 95%, 96%, 97% or 98%, and other specific values in the range can be selected, and are not repeated herein.
Preferably, the fermentation temperature of the liquid fermentation product of bacillus beiLeisi is 25-40 ℃, for example 25 ℃, 28 ℃, 30 ℃, 35 ℃ or 40 ℃, and other specific values in the range can be selected, and are not repeated herein.
Specifically, the fermentation conditions of the bacillus belgii liquid fermentation product are as follows: aerobic at 25-40 deg.C until thallus content is 1011The fermentation was terminated at cfu/mL.
The fermentation conditions of the three fermentations related by the invention have important influence on the product performance, and if the temperature is too low and the dissolved oxygen is insufficient, the bacterial strain is slowly proliferated, so that the fermentation time is long, and the concentration of the bacteria can not meet the requirements; a similar problem occurs with too high a temperature.
In another aspect, the invention provides a soil remediation agent, which comprises the carbon-based composite microbial inoculum and organic materials.
The soil remediation agent can adjust the pH of soil to be neutral, remarkably reduce the salt accumulation of the soil, improve the secondary salinization and acidification conditions of high-load planting soil, increase the organic matter content of the soil and reduce the incidence of the clubroot of the green vegetables in the continuous cropping soil from 60% to about 10%. The quantity of plasmodiophora brassicae in the continuous cropping soil treated by the soil remediation agent disclosed by the invention is obviously reduced, so that the quantity of plasmodiophora brassicae in the root system of the green vegetables in the harvesting period is obviously reduced. The microbial community species diversity in soil and plant root systems is obviously increased after the soil remediation agent is used for treatment.
Preferably, the organic material comprises crop straw and/or green manure. The length of the straw or the green manure is less than or equal to 3-10cm during the specific implementation operation.
Preferably, the crop stalks comprise any one or a combination of at least two of rice straw stalks, wheat straw stalks or corn stalks; the combination of at least two of the above-mentioned materials, such as the combination of rice straw and wheat straw, the combination of wheat straw and corn straw, the combination of rice straw and corn straw, and the like, any other combination mode can be selected, and the details are not repeated herein.
Preferably, the green manure comprises any one or a combination of at least two of broad beans, astragalus sinicus or alfalfa; the combination of at least two of the above-mentioned components, such as the combination of broad bean and milk vetch, the combination of milk vetch and alfalfa, the combination of broad bean and alfalfa, etc., can be selected in any combination manner, and thus, the details are not repeated herein.
Preferably, the mass ratio of the carbon-based composite microbial inoculum to the organic material is 1 (1-10), for example, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10, and the like, and other specific values in the range can be selected, which is not described herein again.
Preferably, the soil is one that has developed clubroot and is intended for continued planting of cruciferous crops.
In yet another aspect, the present invention provides a method of using a soil remediation agent as described above, the method of use comprising: adding a soil repairing agent into the soil, uniformly spreading the soil repairing agent on the field surface, uniformly ploughing the soil and the plough layer soil, flooding or keeping the saturated water content for 2-4 weeks, draining water after the treatment is finished, drying, and then carrying out conventional planting.
Preferably, the soil remediation agent accounts for 0.1-2% of the soil by mass, such as 0.1%, 0.5%, 1% or 2%, and other specific values within the range can be selected, which is not repeated herein.
The soil remediation agent accounts for 0.1-2% of the soil by mass percentage, and the growth of crops is inhibited due to excessively high addition amount. The reason that the growth of subsequent crops is influenced by too high addition amount is as follows: firstly, the heat and substances released by slowly decomposing the residual part of the organic materials in the crop planting period after the soil treatment can influence the crop root development and inhibit the early growth of crops; secondly, a certain amount of nitrogen and nutrients are consumed in the decomposition process of the organic materials, and if the addition amount is large, the soil nutrients required by the growth of crops can be over contended, so that the early growth of the crops is influenced.
Compared with the prior art, the invention has the following beneficial effects:
the carbon-based composite microbial inoculum related by the invention creatively compounds three fermentation products and biomass charcoal, and can quickly start an aerobic and anaerobic treatment process in a field after the three fermentation products are mixed with organic materials for flooding. Wherein, the aspergillus oryzae solid fermentation product and the bacillus beiLeisi liquid fermentation product can quickly start the aerobic decomposition of organic materials in soil, and influence the temperature and oxidation-reduction potential conditions in the treatment process; the zygosaccharomyces rouxii liquid fermentation product participates in anaerobic fermentation; the Bacillus belgii liquid fermentation product also has antagonism to various soil-borne and air-borne plant pathogenic bacteria, and can enhance disease resistance of later-stage crops. The three components have synergistic effect, and when the compound fertilizer is used in a soil remediation agent, the pH value of soil can be adjusted to be neutral under the condition of no pesticide introduction, the soil salt content is reduced, the organic matter content is increased, the incidence rate of the clubroot of the continuous cropping soil green vegetables is reduced, and the incidence rate of the clubroot of the continuous cropping soil green vegetables can be reduced from 60% to about 10%. The quantity of plasmodiophora brassicae in the continuous cropping soil treated by the soil remediation agent disclosed by the invention is obviously reduced, so that the quantity of plasmodiophora brassicae in the root system of the green vegetables in the harvesting period is obviously reduced. The microbial community species diversity in soil and plant root systems is obviously increased after the soil remediation agent is used for treatment. The invention effectively overcomes the defects of high cost of transplanting and serious pollution of chemical prevention and control, has simple operation and high popularization and utilization value.
Detailed Description
To further illustrate the technical means and effects of the present invention, the following further describes the technical solution of the present invention with reference to the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.
The Aspergillus oryzae in the following examples is Aspergillus oryzae (Aspergillus oryzae) S1 deposited in China general microbiological culture Collection center (CGMCC), with the collection number of CGMCC No.16264, the collection date of 2018.8.15, and the collection address of No. 3 North road 1 Chen Xilu.
The Zygosaccharomyces rouxii referred to in the following examples is Zygosaccharomyces rouxii JY1 deposited in the common microorganism center (CGMCC) of the china committee for culture collection of microorganisms, the preservation number is CGMCC No.16260, the preservation date is 2018.8.15, and the preservation address is No. 3 of north chen west road No.1 of the rising area of beijing.
The Bacillus belgii related to the following embodiments is Bacillus belgii (Bacillus velezensis) K3 preserved in common microorganism center (CGMCC) of china culture and management committee for preservation of microorganisms, the preservation number is CGMCC No.19256, the preservation date is 2019.12.30, and the preservation address is No. 3 of north chen west lot No.1 of north township, beijing.
The preparation method of the aspergillus oryzae solid fermentation product related to the following embodiment comprises the following steps: standing Aspergillus oryzae in a fermentation tank at 30 deg.C for 40min, and turning over for 5 min; and continuously fermenting for about 2 weeks until the materials are completely wrapped by white hyphae and the water content is about 40 percent, and finishing the fermentation. The formula of the culture medium is as follows: 10% of bran, 10% of rice hull powder, 70% of rice straw and 10% of soybean meal. The water content was then adjusted to 60%.
The following examples relate to a method for preparing a zygosaccharomyces rouxii liquid fermentation product: culturing the zygosaccharomyces rouxii at 30 deg.C under aerobic condition until thallus content is 1011The fermentation was terminated at cfu/mL. The formula of the culture medium is as follows: 1% of yeast extract, 2% of peptone, 2% of glucose and 95% of water.
The following examples relate to a method for preparing a liquid fermentation product of Bacillus belgii comprising: culturing the Bacillus belgii at 30 deg.C under aerobic condition until the thallus content is 1011The fermentation was terminated at cfu/mL. The formula of the culture medium is as follows: peptone 1%, beef extract 1.5%, sodium chloride 1.5%, water 96%, pH7.2.
The following examples relate to biomass charcoal produced by Shikoku (Shanghai) Biotechnology, Inc., model No.: organic Bamboo Power BBP NO. 6.
Example 1
This example provides a carbon-based composite comprising a solid fermentation product of Aspergillus oryzae, a liquid fermentation product of Zygosaccharomyces rouxii, a liquid fermentation product of Bacillus belgii and biomass carbon, wherein the total viable count of the carbon-based composite is 2.0 × 1010The ratio of the number of viable bacteria of the aspergillus oryzae, the zygosaccharomyces rouxii and the bacillus belgii is 1:2:2, wherein the biomass charcoal accounts for 70% of the carbon-based composite in percentage by mass.
The embodiment also provides a soil remediation agent, wherein the soil remediation agent is a uniformly mixed product of the carbon-based compound and the straw powder, and the mass ratio of the carbon-based compound to the straw powder is 1: 2.
Example 2
This example provides a carbon-based composite comprising a solid fermentation product of Aspergillus oryzae, a liquid fermentation product of Zygosaccharomyces rouxii, a liquid fermentation product of Bacillus belgii and biomass carbon, wherein the total viable count of the carbon-based composite is 3.0 × 1010The ratio of the number of the viable bacteria of the aspergillus oryzae, the zygosaccharomyces rouxii and the bacillus belezii is1:1:1, wherein the biomass charcoal accounts for 60% of the carbon-based composite by mass.
The embodiment also provides a soil remediation agent, wherein the soil remediation agent is a uniformly mixed product of the carbon-based compound and the wheat straw powder, and the mass ratio of the carbon-based compound to the wheat straw powder is 1: 5.
Example 3
This example provides a carbon-based composite comprising a solid fermentation product of Aspergillus oryzae, a liquid fermentation product of Zygosaccharomyces rouxii, a liquid fermentation product of Bacillus belgii and biomass carbon, wherein the total viable count of the carbon-based composite is 1.0 × 109The ratio of the number of viable bacteria of the aspergillus oryzae, the zygosaccharomyces rouxii and the bacillus belgii is 1:1:2, wherein the biomass charcoal accounts for 80% of the carbon-based composite in percentage by mass.
The embodiment also provides a soil repairing agent, wherein the soil repairing agent is a uniformly mixed product of the carbon-based compound and the broad bean powder, and the mass ratio of the carbon-based compound to the broad bean powder is 1: 10.
Example 4
This example provides a carbon-based composite differing from example 1 only in that the total viable count in the carbon-based composite is 1.0X 108All others were kept consistent.
The embodiment also provides a soil remediation agent, wherein the soil remediation agent is a uniformly mixed product of the carbon-based compound and the straw powder, and the mass ratio of the carbon-based compound to the straw powder is 1: 2.
Example 5
This example provides a carbon-based composite differing from example 1 only in that the total viable count in the carbon-based composite is 1.0X 1011All others were kept consistent.
The embodiment also provides a soil remediation agent, wherein the soil remediation agent is a uniformly mixed product of the carbon-based compound and the straw powder, and the mass ratio of the carbon-based compound to the straw powder is 1: 2.
Example 6
This example provides a carbon-based composite that differs from example 1 only in that the ratio of viable count of aspergillus oryzae, zygosaccharomyces rouxii and bacillus beijerinckii is 2:1:1, all else remaining the same.
The embodiment also provides a soil remediation agent, wherein the soil remediation agent is a uniformly mixed product of the carbon-based compound and the straw powder, and the mass ratio of the carbon-based compound to the straw powder is 1: 2.
Example 7
This example provides a carbon-based composite that differs from example 1 only in that the ratio of viable count of aspergillus oryzae, zygosaccharomyces rouxii and bacillus beijerinckii is 1:1:4, all else remaining the same.
The embodiment also provides a soil remediation agent, wherein the soil remediation agent is a uniformly mixed product of the carbon-based compound and the straw powder, and the mass ratio of the carbon-based compound to the straw powder is 1: 2.
Comparative example 1
This comparative example provides a carbon-based composite that differs from example 1 only in that it does not contain the aspergillus oryzae solid fermentate, the ratio of viable count of zygosaccharomyces rouxii to that of bacillus belgii is 1:1, and the rest remains the same.
The comparative example also provides a soil remediation agent, wherein the soil remediation agent is a uniformly mixed product of the carbon-based compound and the straw powder, and the mass ratio of the carbon-based compound to the straw powder is 1: 2.
Comparative example 2
This comparative example provides a carbon-based composite that differs from example 1 only in that it does not contain zygosaccharomyces rouxii, and the ratio of viable count of the aspergillus oryzae solid fermentation product to that of bacillus belezii is 1:2, all other things remaining the same.
The comparative example also provides a soil remediation agent, wherein the soil remediation agent is a uniformly mixed product of the carbon-based compound and the straw powder, and the mass ratio of the carbon-based compound to the straw powder is 1: 2.
Comparative example 3
This comparative example provides a carbon-based composite that differs from example 1 only in that it does not contain bacillus belgii, the ratio of viable count of the aspergillus oryzae solid fermentation to zygosaccharomyces rouxii is 1:2, and the others are identical.
The comparative example also provides a soil remediation agent, wherein the soil remediation agent is a uniformly mixed product of the carbon-based compound and the straw powder, and the mass ratio of the carbon-based compound to the straw powder is 1: 2.
Evaluation test:
the soil remediation agent prepared in the examples 1 to 7 and the comparative examples 1 to 3 is used for preventing and controlling the occurrence of clubroot of the replanted vegetables, and the concrete process is as follows:
the soil remediation agent of each group is added into soil with clubroot disease in an amount of 1%, and is uniformly mixed with plough layer soil, and is watered and kept in a shallow water layer for 2 weeks. Draining water and draining after the treatment is finished, planting green vegetables 'Huawang', setting untreated soil as a contrast, keeping all management measures consistent in the cultivation process, researching the influence of the repairing agent on replanting green vegetables and continuous cropping soil, and obtaining the evaluation results shown in tables 1-4. The number of the rhizomes in the table and the number of the soil rhizomes in the table are determined by a fluorescent quantitative PCR method (instrument model TIB8600), and the analysis of the diversity of the rhizomes and the soil bacteria and the soil fungi is completed by a high-throughput sequencing technology (sequencing platform Illumina Novaseq).
TABLE 1
Group of Incidence (%) Plant height (cm) Chlorophyll content (SPAD) Number of blades (sheet)
Control group 60a 7.6b 41a 9a
Example 1 10b 9.1a 42a 10a
Example 2 10b 9.8a 43b 11b
Example 3 11b 8.8a 41a 10a
Example 4 15b 8.5a 42a 10a
Example 5 9b 10a 43b 11b
Example 6 12b 9.0a 42a 10a
Example 7 10b 8.5a 42a 10a
Comparative example 1 16b 8.5a 42a 10a
Comparative example 2 12b 8.5a 42a 10a
Comparative example 3 14b 8.5a 42a 10a
Note: the lower case letters in the same column indicate significant differences (p < 0.05).
As can be seen from the data in Table 1: the carbon-based composite microbial inoculum related by the invention can obviously reduce the incidence rate of the clubroot of the green vegetables in the continuous cropping soil without introducing pesticides when being used in a soil remediation agent through the synergistic effect of three fermentation products. And the total viable count in the carbon-based composite microbial inoculum meets 1.0 multiplied by 109-3.0×1010When the ratio of the viable count meets the conditions of 1 (1-2) to 1-2, the effect is more obvious.
TABLE 2
Figure GDA0002560419040000141
Note: the lower case letters in the same column indicate significant differences (p < 0.05).
As can be seen from the data in Table 2: the carbon-based composite microbial inoculum related by the invention can obviously reduce the quantity of plasmodiophora brassicae in the roots of the green vegetables and obviously increase the bacterial diversity and the fungal diversity in the roots when being used in a soil remediation agent through the synergistic effect of three fermentation products. And the total viable count in the carbon-based composite microbial inoculum meets 1.0 multiplied by 109-3.0×1010When the ratio of the viable count meets the conditions of 1 (1-2) to 1-2, the effect is more obvious.
TABLE 3
Figure GDA0002560419040000142
Figure GDA0002560419040000151
Note: the lower case letters in the same column indicate significant differences (p < 0.05).
As can be seen from the data in Table 3: the carbon-based composite microbial inoculum related by the invention can obviously adjust the pH of soil to be neutral, reduce the salt content of the soil and increase the organic matter content of the soil when being used in a soil remediation agent through the synergistic effect of three fermentation products. And the total viable count in the carbon-based composite microbial inoculum meets 1.0 multiplied by 109-3.0×1010When the ratio of the viable count meets the conditions of 1 (1-2) to 1-2, the effect is more obvious.
TABLE 4
Figure GDA0002560419040000152
Figure GDA0002560419040000161
Note: the lower case letters in the same column indicate significant differences (p < 0.05).
From the data in table 4, it can be seen that: the carbon-based composite microbial inoculum related by the invention can obviously reduce the number of soil plasmodiophora brassicae and obviously increase the diversity of soil bacteria and fungi when being used in a soil remediation agent through the synergistic effect of three fermentation products. And the total viable count in the carbon-based composite microbial inoculum meets 1.0 multiplied by 109-3.0×1010When the ratio of the viable count meets the conditions of 1 (1-2) to 1-2, the effect is more obvious.
The applicant states that the present invention is described by the above embodiments and the application of the carbon-based composite microbial inoculum of the present invention, but the present invention is not limited to the above embodiments, that is, the present invention is not meant to be implemented only by the above embodiments. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.

Claims (16)

1. The carbon-based compound microbial inoculum for reducing the incidence rate of clubroot of continuous cropping soil green vegetables is characterized by comprising an aspergillus oryzae solid fermentation product, a zygosaccharomyces rouxii liquid fermentation product, a bacillus beijerinckii liquid fermentation product and biomass charcoal;
the ratio of the number of the live bacteria of the aspergillus oryzae, the zygosaccharomyces rouxii and the bacillus belgii is 1 (1-2) to 1-2;
the total viable count of Aspergillus oryzae, Zygosaccharomyces rouxii and Bacillus belgii in the carbon-based composite microbial agent is 1.0 × 109-3.0×1010cfu/mL；
The Aspergillus oryzae is Aspergillus oryzae (Aspergillus oryzae)Asperigillus oryzae) S1, the preservation number is CGMCC No. 16264;
the zygosaccharomyces rouxii is zygosaccharomyces rouxii (A)Zygosaccharomyces rouxii) JY1 with preservation number CGMCC No. 16260;
the Bacillus belgii is Bacillus belgii (Bacillus belgii)Bacillus velezensis) K3 with the preservation number of CGMCC No. 19256.
2. The charcoal-based composite microbial inoculum according to claim 1, wherein the mass percentage of the biomass charcoal in the charcoal-based composite microbial inoculum is 60-80%.
3. The carbon-based composite microbial inoculum of claim 1, wherein the fermentation medium formula of the aspergillus oryzae solid fermentation product comprises the following components in percentage by weight: 2-8% of bran, 2-8% of rice hull powder, 25-35% of rice straw, 2-6% of soybean flour and 50-70% of water.
4. The carbon-based composite microbial inoculum according to claim 1, wherein the fermentation starting temperature of the aspergillus oryzae solid fermentation product is 20-40 ℃.
5. The carbon-based composite microbial inoculant according to claim 1, wherein the fermentation medium formula of the zygosaccharomyces rouxii liquid fermentation product comprises the following components in percentage by weight: 1-5% of yeast extract, 1-3% of peptone, 1-3% of glucose and 90-97% of water.
6. The carbon-based composite microbial inoculant according to claim 1, wherein the fermentation temperature of the zygosaccharomyces rouxii liquid fermentation product is 25-40 ℃.
7. The carbon-based composite microbial inoculum according to claim 1, wherein the fermentation medium formula of the Bacillus belgii liquid fermentation product comprises the following components in percentage by weight: peptone 1-3%, beef extract 0.5-2.5%, sodium chloride 0.5-2.5% and water 93-98%.
8. The carbon-based composite microbial agent according to claim 1, wherein the fermentation temperature of the Bacillus belgii liquid fermentation product is 25 to 40 ℃.
9. A soil remediation agent comprising the carbon-based complex inoculant of claim 1 and an organic material.
10. The soil remediation agent of claim 9 wherein the organic material comprises crop straw and/or green manure.
11. The soil remediation agent of claim 10 wherein the crop stalks comprise any one or a combination of at least two of rice straw, wheat straw, or corn stover.
12. The soil remediation agent of claim 10 wherein the green manure comprises any one or a combination of at least two of fava beans, milk vetch or alfalfa.
13. The soil remediation agent of claim 9 wherein the mass ratio of the carbon-based complex inoculant to the organic material is 1 (1-10).
14. The soil remediation agent of claim 9 wherein the soil is one that has developed clubroot and is intended for continued growth of cruciferous crops.
15. The method of using a soil remediation agent of claim 9, the method of using comprising: adding a soil repairing agent into the soil, uniformly spreading the soil repairing agent on the field surface, uniformly ploughing the soil and the plough layer soil, flooding or keeping the saturated water content for 2-4 weeks, draining water after the treatment is finished, drying, and then carrying out conventional planting.
16. The soil remediation agent of claim 15 wherein the soil remediation agent is present in an amount of 0.1% to 2% by weight of the soil.
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