CN112391302A

CN112391302A - Solid compound microbial agent for soil remediation and preparation method thereof

Info

Publication number: CN112391302A
Application number: CN201910738207.1A
Authority: CN
Inventors: 周廷霖
Original assignee: Yangtian Biotechnology Jiangsu Co ltd
Current assignee: Yangtian Biotechnology Jiangsu Co ltd
Priority date: 2019-08-12
Filing date: 2019-08-12
Publication date: 2021-02-23

Abstract

The application discloses a solid compound microbial agent for soil remediation and a preparation method thereof. The microbial agent comprises the following components in percentage by weight: 35 to 45 percent of radioactive ray bacteria, 3 to 7 percent of bacillus subtilis, 3 to 7 percent of mucor, 3 to 7 percent of rhizopus, 3 to 7 percent of cephalospora, 3 to 7 percent of hansenula, 3 to 7 percent of ascomycetes, 5 to 15 percent of aspergillus niger, 5 to 15 percent of penicillium and 5 to 15 percent of neurospora tetraspora. The method solves the technical problem that the prior art is poor in soil remediation capability due to the fact that the combination of more different fungi, actinomycetes and bacillus subtilis is not wide, more fungi are single strains, or the combination of several bacillus subtilis is not wide.

Description

Solid compound microbial agent for soil remediation and preparation method thereof

Technical Field

The invention relates to the technical field of biochemical engineering, and particularly relates to a solid compound microbial agent for soil remediation and a preparation method thereof.

Background

In agricultural cultivation environments, due to the long-term use of various chemical agents, such as fertilizers, herbicides, pesticides, bactericides and the like, the microbial balance of soil is damaged, and further, the problems of reduction of the absorption and utilization rate of plant fertilizers, soil acidification, soil diseases and insect pests and the like are caused.

Aiming at the problem that the prior art in the related technology does not widely combine more different fungi, actinomycetes and bacillus subtilis, more single strains or the combination of several bacillus subtilis cause poor soil remediation capability, an effective solution is not provided at present.

Disclosure of Invention

The application mainly aims to provide a solid compound microbial agent for soil remediation and a preparation method thereof, so as to solve the problem that the soil remediation capability is poor due to the fact that the prior art does not widely combine more different combinations of fungi, actinomycetes and bacillus subtilis, and more single strains or the combination of several bacillus subtilis.

In order to achieve the above objects, according to one aspect of the present application, there is provided a solid complex microbial inoculant for soil remediation and a method for preparing the same.

The solid compound microbial agent for soil remediation comprises the following components in percentage by weight: 35 to 45 percent of radioactive ray bacteria, 3 to 7 percent of bacillus subtilis, 3 to 7 percent of mucor, 3 to 7 percent of rhizopus, 3 to 7 percent of cephalospora, 3 to 7 percent of hansenula, 3 to 7 percent of ascomycetes, 5 to 15 percent of aspergillus niger, 5 to 15 percent of penicillium and 5 to 15 percent of neurospora tetraspora.

Further, the method comprises the following steps: the content of radioactive ray bacteria is 40%, the content of bacillus subtilis is 5%, the content of mucor is 5%, the content of rhizopus is 5%, the content of cephalospora is 5%, the content of hansenula, the content of ascomycetes is 5%, the content of aspergillus niger is 10%, the content of penicillium and the content of neurospora tetraspora are 10%.

In order to achieve the above object, according to another aspect of the present application, there is provided a method for preparing a solid complex microbial inoculant.

A preparation method of a solid compound microbial agent comprises the following steps:

step 1), adding a mixed solution of a milt extract, asparagine, protein and glycerol into a liquid culture medium composed of phosphoric acid, manganese sulfate and ammonium sulfate, and culturing for 24 hours in a stainless steel sterile liquid fermentation tank at a temperature of 30-37 ℃ and a pH value of 4-6;

step 2) before adding solid for fermentation, all materials are firstly subjected to sterilization treatment, and are added into a solid sterile stainless steel fermentor after the aseptic state is ensured, the solid fermentation product takes vermiculite as a carrier, molasses as a main carbon source, soybean meal, yeast powder, oyster shell powder and chitosan;

step 3) after liquid culture for 24 hours, sampling and detecting the concentration of the thalli, and transferring the bacterial liquid into a sterile stainless steel fermentor for culture after the growth condition of the thalli is confirmed;

step 4), when the solid fermentation device is used for fermentation, controlling the temperature to be 30-37 ℃, shaking at 120rpm, and continuously culturing and fermenting for 7 days;

step 5) periodically sampling and detecting the activity, the spore transfer condition and the pH value parameter of the bacterial colony during the culture and growth period;

step 6), after fermentation is finished, cooling and drying at low temperature of 40 ℃ for 2-4 days to remove water and keep high survival rate of the thalli;

and 7) after drying and cooling, packaging into a finished product, and finishing the production process.

Further, in the step 1, the content of phosphoric acid is 4%, the content of manganese sulfate is 1%, the content of ammonium sulfate is 4%, the content of the milt extract is 5%, the content of asparagine is 5%, the content of protein is 5%, and the content of glycerol is 5%.

Further, in the step 2, the content of the vermiculite is 25% and the content of the molasses is 2.5%.

Further, the compound microbial agent is cultured by two-stage fermentation.

In the embodiment of the application, a mode of combining various bacteria and fungi together is adopted, and each strain is formed by different proportion proportions, so that the purpose of forming the compound microbial agent is achieved, the technical effect of improving the soil remediation capability is achieved, and the technical problem that the soil remediation capability is poor due to the fact that the prior art does not have the wide combination of more different fungi, actinomycetes and bacillus subtilis, more single strains or the combination of several bacillus subtilis is solved.

Drawings

FIG. 1 is a diagram showing the use of a solid complex microbial inoculant according to the present invention for soil remediation.

Detailed Description

In order to make the technical solutions in the embodiments of the present application better understood, the technical solutions in the embodiments of the present application are clearly and completely described, and it is obvious that the described embodiments are only some embodiments of the present application, not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

It is noted that the terms first, second and the like in the description and in the claims of the present application are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It should be understood that the data so used may be interchanged under appropriate circumstances such that embodiments of the application described herein may be used. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.

Moreover, some of the above terms may be used to indicate other meanings besides the orientation or positional relationship, for example, the term "on" may also be used to indicate some kind of attachment or connection relationship in some cases. The specific meanings of these terms in the present invention can be understood by those skilled in the art as appropriate.

Furthermore, the terms "mounted," "disposed," "provided," "connected," and "sleeved" are to be construed broadly. For example, it may be a fixed connection, a removable connection, or a unitary construction; can be a mechanical connection, or an electrical connection; may be directly connected, or indirectly connected through intervening media, or may be in internal communication between two devices, elements or components. The specific meanings of the above terms in the present invention can be understood by those of ordinary skill in the art according to specific situations.

It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present application will be described in detail with reference to examples.

Example 1:

a solid compound microbial agent for soil remediation comprises the following components in percentage by weight: 35 to 45 percent of radioactive ray bacteria, 3 to 7 percent of bacillus subtilis, 3 to 7 percent of mucor, 3 to 7 percent of rhizopus, 3 to 7 percent of cephalospora, 3 to 7 percent of hansenula, 3 to 7 percent of ascomycetes, 5 to 15 percent of aspergillus niger, 5 to 15 percent of penicillium and 5 to 15 percent of neurospora tetraspora.

Wherein, the actinomycetes are prokaryotes which mainly grow in mycelial form and propagate by spores and have stronger terrestrial property, and have the functions of weeding and resisting parasites;

the bacillus subtilis spores exist in the soil remediation agent in a dormant state, and the spore-form bacteria have the characteristics of high temperature resistance, salt resistance and drying resistance; when the compound microbial agent is used in soil, bacterial spores rapidly grow and reproduce under proper moisture and temperature, the produced extracellular enzyme can decompose organic matters in the soil and directly provide nutrition for crops, and metabolites of the extracellular enzyme have the effect of stimulating the metabolites, can effectively prevent or reduce diseases and insect hazards, and is beneficial to the robust growth of the crops;

the yeast flora can synthesize active substances for promoting root growth and cell division, and provide important nutrition guarantee for substrates (food) required for promoting proliferation of other effective microorganisms;

the aspergillus niger can crack macromolecular organic matters and indissolvable inorganic matters, is convenient for crops to absorb and utilize, improves the soil structure, enhances the soil fertility and improves the crop yield;

in the standard solution of mercury and lead, the compound microbial agent is used to perform microbial daughter-in-law test, and mercury ions (Hg) are found²⁺) And lead ion (Pb)²⁺) The adsorption rate of (A) is 80-97%;

the organochlorine pesticide usually remains in soil for a long time due to the problems of low volatility and difficult decomposition, thereby causing environmental pollution; the compound microbial agent is applied to soil with pesticide DDT and BHC, which can accelerate DDT decomposition to be degraded into other low-toxicity derivatives or metabolites, and then the DDT is degraded and decomposed to remove, and the adsorption and decomposition rate after use reaches 90-100%.

From the above description, it can be seen that the following technical effects are achieved by the present application:

The invention also has the following beneficial effects:

1. solving the problem of residues of various heavy metals or organic solvents in soil, e.g.

-pesticide residues (DDT-dichlorophenyl trichloroethane & BHC-hexachlorocyclohexane, etc.);

heavy metal residues (such as mercury and lead, etc.);

2. improve soil fertility and improve fertilizer utilization rate

-dissolving phosphorus: releasing the phosphorus element fixed in the soil;

-potassium solubilisation: releasing the potassium element fixed in the soil;

3. promoting plant growth

-binding to the plant root system, increasing the plant root uptake area;

secretion of organic acids, amylases, proteases, indoleacetic acid, etc. to promote plant absorption;

4. reduction of soil diseases and insect pests

Inhibition of soil spoilage bacteria growth such as: inhibiting soil nematodes;

5. improving soil properties

-increasing soil water retention, increasing soil aeration;

-loosening soil hardening problems;

-improving soil acidification.

Example 2:

a solid complex microbial inoculant for soil remediation, comprising: the content of radioactive ray bacteria is 40%, the content of bacillus subtilis is 5%, the content of mucor is 5%, the content of rhizopus is 5%, the content of cephalospora is 5%, the content of hansenula, the content of ascomycetes is 5%, the content of aspergillus niger is 10%, the content of penicillium and the content of neurospora tetraspora are 10%.

in the standard solution of mercury and lead, the compound microbial agent is used to perform microbial daughter-in-law test, and mercury ions (Hg) are found²⁺) And lead ion (Pb)²⁺) The adsorption rate of (2) is above 90% (97% and 92%, respectively);

the organochlorine pesticide usually remains in soil for a long time due to the problems of low volatility and difficult decomposition, thereby causing environmental pollution; the compound microbial agent is applied to soil with pesticide DDT and BHC, which can accelerate DDT decomposition to be degraded into other low-toxicity derivatives or metabolites, and then is degraded and decomposed to remove, and the adsorption and decomposition rate after use reaches 100%.

The optimal use effect of the compound microbial agent can be realized through the proportion.

The embodiment also provides a preparation method of the solid compound microbial agent;

example 3: the preparation method comprises the following steps:

The vermiculite is volcanic rock, and is obtained by high-temperature calcination at 1400-2200 ℃, drying and destruction; the characteristics of vermiculite porosity and thermal expansion can provide a good microbial growth environment, so that the technology uses the vermiculite as a carrier to jointly culture and combine multiple microorganisms without generating antagonism; the microbial agent cultured in the way can improve the soil acidification problem, increase the permeability of soil, improve the physical properties of soil, decompose soil toxins (such as herbicides, pesticides and the like) and improve the absorption and utilization rate of soil fertilizers when being applied to the ground due to the diversified microbial composition.

By the preparation method, the compound microbial agent can be prepared and obtained, and the success rate is over 90%.

Example 4:

step 1), adding a mixed solution of a milt extract (5%), asparagine (5%), protein (5%) and glycerol (5%) into a liquid culture medium composed of phosphoric acid (4% solution), manganese sulfate (1%) and ammonium sulfate (1% solution), and culturing for 24 hours in a stainless steel sterile liquid fermentation tank at a temperature of 30-37 ℃ and a pH value of 4-6;

step 2) before adding solid for fermentation, all materials are firstly subjected to sterilization treatment, and are added into a solid sterile stainless steel fermentor after being ensured to be in a sterile state, the solid fermentation product takes vermiculite (25%) as a carrier, molasses (2.5%) as a main carbon source, soybean meal, yeast powder, oyster shell powder and chitosan (22.5);

By the preparation method, the compound microbial agent can be prepared and obtained, and the success rate is over 98%.

Example 5:

step 1), performing two-stage fermentation culture on the compound microbial agent;

step 2), adding a mixed solution of a milt extract (5%), asparagine (5%), protein (5%) and glycerol (5%) into a liquid culture medium composed of phosphoric acid (4% solution), manganese sulfate (1%) and ammonium sulfate (1% solution), and culturing for 24 hours in a stainless steel sterile liquid fermentation tank at a temperature of 30-37 ℃ and a pH value of 4-6;

step 3) before adding the solid for fermentation, all materials are firstly subjected to sterilization treatment, and are added into a solid sterile stainless steel fermentor after being ensured to be in a sterile state, the solid fermentation product takes vermiculite (25%) as a carrier, molasses (2.5%) as a main carbon source, soybean meal, yeast powder, oyster shell powder and chitosan (22.5);

step 4), after liquid culture is carried out for 24 hours, sampling and detecting the concentration of thalli, and transferring the bacterial liquid into a sterile stainless steel fermentor for culture after the growth condition of the thalli is confirmed;

step 5) when the solid fermentation device is used for fermentation, controlling the temperature to be 30-37 ℃, shaking at 120rpm, and continuously culturing and fermenting for 7 days;

step 6) periodically sampling and detecting the activity, the spore transfer condition and the pH value parameter of the bacterial colony during the culture and growth period;

step 7), after fermentation is finished, cooling and drying at low temperature of 40 ℃ for 2-4 days to remove water and keep high survival rate of the thalli;

and 8) after drying and cooling, packaging into a finished product, and finishing the production process.

Firstly, liquid fermentation culture is carried out, so that the microorganism can achieve the effect of rapid proliferation in the liquid culture.

The test results according to the examples are shown in the following table:

table 1:

table 2:

from the above table, it can be seen that: after the composite microbial agent is used in soil, the plant height of crops and the yield and quality of melons and fruits are obviously improved.

From the above table, it can be seen that: the compound microbial agent does not influence the activity of bacterial colonies, thereby improving the soil improving capability.

The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. The solid compound microbial agent for soil remediation is characterized by comprising the following components in percentage by weight: 35 to 45 percent of radioactive ray bacteria, 3 to 7 percent of bacillus subtilis, 3 to 7 percent of mucor, 3 to 7 percent of rhizopus, 3 to 7 percent of cephalospora, 3 to 7 percent of hansenula, 3 to 7 percent of ascomycetes, 5 to 15 percent of aspergillus niger, 5 to 15 percent of penicillium and 5 to 15 percent of neurospora tetraspora.

2. The solid complex microbial inoculant for soil remediation according to claim 1 comprising: the content of radioactive ray bacteria is 40%, the content of bacillus subtilis is 5%, the content of mucor is 5%, the content of rhizopus is 5%, the content of cephalospora is 5%, the content of hansenula, the content of ascomycetes is 5%, the content of aspergillus niger is 10%, the content of penicillium and the content of neurospora tetraspora are 10%.

3. A preparation method of a solid compound microbial agent is characterized by comprising the following steps:

4. The method according to claim 3, wherein in step 1, the phosphoric acid content is 4% solution, the manganese sulfate content is 1%, the ammonium sulfate content is 4% solution, the milt extract content is 5%, the asparagine content is 5%, the protein content is 5%, and the glycerol content is 5%.

5. The method for preparing a solid complex microbial inoculant according to claim 3, wherein in step 2 the vermiculite content is 25% and the molasses content is 2.5%.

6. The solid complex microbial inoculant according to claim 1, wherein the complex microbial inoculant is cultured by two-stage fermentation.