CN113337419B - Preparation method of microbial agent for soil acidification areas - Google Patents
Preparation method of microbial agent for soil acidification areas Download PDFInfo
- Publication number
- CN113337419B CN113337419B CN202110453802.8A CN202110453802A CN113337419B CN 113337419 B CN113337419 B CN 113337419B CN 202110453802 A CN202110453802 A CN 202110453802A CN 113337419 B CN113337419 B CN 113337419B
- Authority
- CN
- China
- Prior art keywords
- strains
- microbial agent
- screening
- soil
- liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
- C09K17/18—Prepolymers; Macromolecular compounds
- C09K17/32—Prepolymers; Macromolecular compounds of natural origin, e.g. cellulosic materials
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/48—Holding appliances; Racks; Supports
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/50—Means for positioning or orientating the apparatus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/12—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by pressure
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
- C12M37/04—Seals
-
- 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
- C12N1/16—Yeasts; Culture media therefor
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Sustainable Development (AREA)
- Molecular Biology (AREA)
- Clinical Laboratory Science (AREA)
- Materials Engineering (AREA)
- Soil Sciences (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Botany (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a preparation method of a microbial agent for a soil acidification area, which belongs to the technical field of microbial agents, creatively increases a process of screening and culturing strains by utilizing soil of a target area, samples are taken in the target area before the microbial agent is prepared, impurities of soil samples are removed, sterilization and water addition are carried out for mixing to prepare screening liquid, and then the strains are screened by the screening liquid to obtain high-quality strains which can survive in the screening liquid, namely the strains which can adapt to the soil environment of the target area, so that the survival rate of microorganisms in the prepared microbial agent can be greatly improved when the microbial agent is used in the target area, the effect of the microbial agent is effectively improved, the practicability is strong, and the microbial collecting balls are arranged for collecting the high-quality strains, can be used for quickly and effectively collecting the screened high-quality strains based on the chemotaxis of microorganisms, and can effectively improve the preparation efficiency.
Description
Technical Field
The invention relates to the technical field of microbial agents, in particular to a preparation method of a microbial agent for soil acidification areas.
Background
The microbial agent is a living microbial agent prepared by processing fermentation liquor of adsorbed bacteria by using porous substances as adsorbents (such as turf and vermiculite) after industrial production and propagation of target microorganisms (effective bacteria). The microbial agent is used for dressing seeds or dipping roots, has the effects of directly or indirectly improving soil, recovering soil fertility, preventing soil-borne diseases, maintaining balance of rhizosphere microbial systems, degrading toxic substances and the like, and can improve the yield of agricultural products, improve the quality of the agricultural products, reduce the dosage of chemical fertilizers, reduce the cost, improve the soil and protect the ecological environment by proper use of the agricultural microbial agent.
When the microbial agent in the prior art is used in areas with serious soil acidification, the soil is acidic, so that microorganisms in the microbial agent easily die in a large scale because the microbial agent cannot adapt to severe soil environments, and the effect and the action of the microbial agent can be greatly reduced. Therefore, we propose a preparation method of microbial agent for soil acidification area.
Disclosure of Invention
1. Technical problem to be solved
Aiming at the problems existing in the prior art, the invention aims to provide a preparation method of a microbial agent for a soil acidification area, which can creatively increase the procedures of screening and culturing strains by utilizing soil of a target area, sampling is firstly carried out in the target area before the microbial agent is prepared, the soil sample is subjected to impurity removal, sterilization and water addition and mixing to prepare screening liquid, and then the strains are screened by the screening liquid, so that high-quality strains which can survive in the screening liquid, namely the strains which can adapt to the soil environment of the target area, are screened, thereby greatly improving the survival rate of the microorganisms in the prepared microbial agent when the microbial agent is used in the target area, further effectively improving the effect of the microbial agent, having strong practicability, being provided with microorganism collecting balls for collecting the high-quality strains, and being capable of quickly and effectively collecting the screened high-quality strains based on the chemotaxis of the microorganisms, thereby effectively improving the preparation efficiency.
2. Technical proposal
In order to solve the problems, the invention adopts the following technical scheme.
The preparation method of the microbial agent for the soil acidification areas comprises the following steps:
s1, taking a proper amount of in-situ soil in a target area as a soil sample, removing impurities and garbage in the soil sample, and obtaining a impurity-removed soil sample;
s2, performing high-temperature sterilization on the impurity-removed soil sample to obtain a sterilized impurity-removed soil sample;
s3, fully mixing and stirring the sterilized impurity-removed soil sample and water, naturally precipitating, and taking supernatant as screening liquid;
s4, pouring the screening liquid into a screening box, then adding beneficial microorganism strains and a liquid culture medium into the screening box, mixing to form strain screening culture liquid, and carrying out directional screening on strains for 2-3d;
s5, placing the microorganism collecting balls into a screening box, and collecting beneficial microorganism strains which survive in the screening box;
s6, taking out the microorganism strains collected by the microorganism collecting balls to obtain high-quality strains, and carrying out propagation on the high-quality strains;
s7, fermenting the high-quality strain subjected to propagation in the step S6 to obtain bacterial liquid;
and S8, uniformly mixing the bacterial liquid in the step S7 with the fermentation material, and then drying in an oven to obtain the microbial agent.
Further, the high-temperature sterilization temperature in the step S2 is 90-130 ℃, the sterilization time is 20-40min, and the mass ratio of the sterilization impurity-removing soil sample to the water in the step S3 is 1:1-2.
Further, the beneficial microorganism strain in the S4 is one or more of lactobacillus, rhizobium, bacillus and saccharomycetes, and the volume ratio of the beneficial microorganism to the culture medium to the screening liquid is 1:20-30:50-100.
Further, the fermentation material in the step S8 is formed by uniformly mixing bean flour and bran according to the volume ratio of 1:4, and the volume ratio of the fermentation material to the bacterial liquid in the step S8 is 1:2-3.
Further, microorganism collecting ball includes hollow spherical shell, install accuse water semipermeable membrane is inlayed to the bottom of hollow spherical shell, the chemotactic ball that two symmetries set up of fixedly connected with on the inner wall of hollow spherical shell, the middle part of hollow spherical shell is provided with elastic air bag, all fixedly connected with gangbar on the outer wall of elastic air bag both sides, the through-hole has all been seted up on the outer wall of hollow spherical shell both sides, two the gangbar keep away from the equal fixedly connected with of one end of elastic air bag and through-hole assorted hole stopper, two the hole stopper is pegged graft with two through-holes respectively.
Furthermore, the chemotactic ball is filled with glucose solution, a plurality of attached villi are fixedly connected to the outer wall of the chemotactic ball, the chemotactic ball is made of semi-permeable membrane materials, membrane holes on the chemotactic ball are smaller than glucose molecules, after the microorganism collecting ball is placed in the screening box, microorganism strains which survive after being screened by screening liquid, namely high-quality strains which can be in a proper soil acidification environment, can move towards the chemotactic ball based on the attractiveness of the glucose solution to the microorganisms, and the through holes enter the hollow spherical shell, so that the collection of the high-quality strains is completed.
Further, the inner wall of the elastic air bag is provided with a shaping net, the top end of the hollow spherical shell is provided with a discharge hole, the middle part of the discharge hole is penetrated and provided with an air duct, the air duct sequentially penetrates through the elastic air bag and the outer wall of the top end of the shaping net and extends into the shaping net, the outer walls of the two sides of the air duct are fixedly connected with support rods, one end of each support rod, which is far away from the air duct, is fixedly connected with the inner wall of the hollow spherical shell, the upper part of the air duct is provided with an elastic air bag, the top end of the elastic air bag is communicated with an air suction check valve, the bottom end of the elastic air bag is communicated with an air duct, one end of the air duct, which is far away from the elastic air bag, is communicated with the air duct, the outer wall of one side of the air duct is provided with an air release valve, through the arrangement of the elastic air bag, the linkage rod, the hole plug, the air guide pipe, the elastic air bag, the air suction one-way valve and the air outlet one-way valve, after the microorganism collecting ball is placed into the screening box, the elastic air bag is repeatedly extruded, air is pumped into the shaping net, so that the shaping net is expanded, the hole plug is driven to move towards the outside of the hollow ball shell until the hole plug is separated from the through hole, the through hole is conducted, and then high-quality strains can move towards the chemotactic ball to be close, after collection is completed, the air release valve can be opened to prevent the shaping net, so that the hole plug blocks the through hole again, then the microorganism collecting ball is lifted to control water, water in the hollow ball shell flows out through the water control semi-permeable membrane, then a proper amount of liquid culture medium is poured into the hollow ball shell, and after the microorganism collecting ball is inverted, high-quality strains can be poured out through the discharge hole, and then the high-quality strains are taken out.
Furthermore, the outer wall of the air duct is fixedly sleeved with the guide hopper, so that the air bag can be prevented from being polluted and elastically beaten when high-quality strains are poured out, and the outer wall of the hollow spherical shell is fixedly connected with two symmetrically arranged holding handles, so that the microorganism collecting balls can be conveniently lifted and inverted.
Further, all be provided with on the inner wall of screening case both sides with hole stopper assorted locating piece, when extrusion elasticity is beaten the gasbag and is made the hole stopper to outside the hollow spherical shell remove, can make the hole stopper remove to stop extrusion elasticity again and beat the gasbag after offing with the locating piece, on the one hand, can play a location effect, on the other hand, can prevent that microorganism collecting ball from empting, after putting into the screening case with microorganism collecting ball in S5, the liquid level height of bacterial screening culture solution is not less than the bottom of hole stopper, and the liquid level height is too low can lead to the unable entering hollow spherical shell of high-quality bacterial through the through-hole.
3. Advantageous effects
Compared with the prior art, the invention has the advantages that:
(1) The method creatively increases the procedures of screening and culturing strains by utilizing the soil of a target area, samples the target area before preparing the microbial inoculum, removes impurities from soil samples, sterilizes, adds water and mixes the soil samples to prepare screening liquid, screens the strains by the screening liquid to screen out high-quality strains which can survive in the screening liquid, namely the strains which can adapt to the soil environment of the target area, so that when the prepared microbial inoculum is used in the target area, the survival rate of microorganisms in the microbial inoculum can be greatly improved, the effect of the microbial inoculum is effectively improved, the practicability is strong, and the microbial collection balls for collecting the high-quality strains are arranged, can quickly and effectively collect the screened high-quality strains based on chemotaxis of the microorganisms, and can effectively improve the preparation efficiency.
(2) The chemotactic ball is filled with glucose solution, a plurality of attached villi are fixedly connected to the outer wall of the chemotactic ball, the chemotactic ball is made of semi-permeable membrane materials, membrane holes on the chemotactic ball are smaller than glucose molecules, microorganism strains which survive after being screened by screening liquid are selected in a screening box, namely high-quality strains which can be in a proper soil acidification environment, can move to the chemotactic ball to be close to the chemotactic ball based on the attractiveness of the glucose solution, and the through holes enter the hollow spherical shell, so that the collection of the high-quality strains is completed.
(3) Through the arrangement of the elastic air bag, the linkage rod, the hole plug, the air guide pipe, the elastic air bag, the air suction one-way valve and the air outlet one-way valve, after the microorganism collecting ball is placed into the screening box, the elastic air bag is repeatedly extruded, air is pumped into the shaping net, so that the shaping net is expanded, the hole plug is driven to move towards the outside of the hollow ball shell until the hole plug is separated from the through hole, the through hole is conducted, and then high-quality strains can move towards the chemotactic ball to be close, after collection is completed, the air release valve can be opened to prevent the shaping net, so that the hole plug blocks the through hole again, then the microorganism collecting ball is lifted to control water, water in the hollow ball shell flows out through the water control semi-permeable membrane, then a proper amount of liquid culture medium is poured into the hollow ball shell, and after the microorganism collecting ball is inverted, high-quality strains can be poured out through the discharge hole, and then the high-quality strains are taken out.
(4) The outer wall of the air duct is fixedly sleeved with the guide hopper, so that the air bag is prevented from being blown off by the elasticity when high-quality strains are poured out, and the outer wall of the hollow spherical shell is fixedly connected with two symmetrically arranged holding handles, so that the microorganism collecting balls can be conveniently lifted and inverted.
(5) The inner walls of the two sides of the screening box are provided with positioning blocks matched with the hole plugs, when the elastic punching bags are extruded to enable the hole plugs to move out of the hollow spherical shell, the elastic punching bags can be stopped from being extruded after the hole plugs are moved to abut against the positioning blocks, on one hand, a positioning effect can be achieved, on the other hand, the microorganism collecting balls can be prevented from toppling over, after the microorganism collecting balls are placed in the screening box in S5, the liquid level of the strain screening culture solution is not lower than the bottom end of the hole plugs, and high-quality strains cannot enter the hollow spherical shell through the through holes due to the fact that the liquid level is too low.
Drawings
FIG. 1 is a flow chart of the preparation of the present invention;
FIG. 2 is a schematic cross-sectional structural view of a microorganism-collecting pellet of the present invention;
FIG. 3 is a schematic cross-sectional view of the elastic balloon of the present invention;
FIG. 4 is a schematic cross-sectional view of the present invention at the guide funnel;
FIG. 5 is a schematic cross-sectional view of a chemotactic ball of the present invention;
FIG. 6 is a schematic view showing the structure of the microorganism-collecting pellet of the present invention when it is put into a screening box.
The reference numerals in the figures illustrate:
101. a hollow spherical shell; 102. a water control semipermeable membrane; 103. a chemotactic ball; 104. an elastic air bag; 105. shaping net; 106. a linkage rod; 107. a hole plug; 108. a discharge hole; 109. an air duct; 110. a support rod; 111. elastically inflating an air bag; 112. an air suction one-way valve; 113. an air outlet one-way valve; 114. a release valve; 115. a diversion bucket; 116. holding the handle; 117. a glucose solution; 118. attaching fluff; 002. a screening box; 201. and (5) positioning blocks.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention; it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments, and that all other embodiments obtained by persons of ordinary skill in the art without making creative efforts based on the embodiments in the present invention are within the protection scope of the present invention.
In the description of the present invention, it should be noted that the positional or positional relationship indicated by the terms such as "upper", "lower", "inner", "outer", "top/bottom", etc. are based on the positional or positional relationship shown in the drawings, are merely for convenience of describing the present invention and simplifying the description, and do not indicate or imply that the apparatus or elements referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the present invention, it should be noted that, unless explicitly specified and limited otherwise, the terms "mounted," "configured to," "engaged with," "connected to," and the like are to be construed broadly, and may be either fixedly connected, detachably connected, or integrally connected, for example; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art.
Example 1:
referring to fig. 1 and 6, a preparation method of a microbial agent for a soil acidification area comprises the following steps:
s1, taking a proper amount of in-situ soil in a target area as a soil sample, removing impurities and garbage in the soil sample, and obtaining a impurity-removed soil sample;
s2, performing high-temperature sterilization on the impurity-removed soil sample to obtain a sterilized impurity-removed soil sample, wherein the sterilization temperature is 90-130 ℃ and the sterilization time is 20-40min;
s3, fully mixing and stirring the sterilized and impurity-removed soil sample and water, wherein the mass ratio of the sterilized and impurity-removed soil sample to the water is 1:1-2, taking supernatant after natural precipitation as screening liquid;
s4, pouring the screening liquid into a screening box 002, then adding beneficial microorganism strains and a liquid culture medium into the screening box 002, mixing to form strain screening culture liquid, and directionally screening strains, wherein the beneficial microorganism strains are one or more of lactobacillus, rhizobium, bacillus and saccharomycetes, and the volume ratio of the beneficial microorganisms to the culture medium to the screening liquid is 1:20-30:50-100, wherein the screening time is 2-3d;
s5, placing the microorganism collecting balls into a screening box 002, and collecting beneficial microorganism strains which survive in the screening box 002;
s6, taking out the microorganism strains collected by the microorganism collecting balls to obtain high-quality strains, and carrying out propagation on the high-quality strains;
s7, fermenting the high-quality strain subjected to propagation in the step S6 to obtain bacterial liquid;
s8, uniformly mixing the bacterial liquid in the S7 with a fermentation material, wherein the fermentation material is formed by uniformly mixing bean flour and bran according to the volume ratio of 1:4, and the volume ratio of the fermentation material to the bacterial liquid is 1:2-3, then placing the mixture in an oven for drying, and obtaining the microbial agent.
Referring to fig. 2 and 5, the microorganism collecting ball includes a hollow ball shell 101, a water-controlling semipermeable membrane 102 is inlaid at the bottom end of the hollow ball shell 101, two symmetrically arranged chemotactic balls 103 are fixedly connected to the inner wall of the hollow ball shell 101, an elastic air bag 104 is arranged in the middle of the hollow ball shell 101, linkage rods 106 are fixedly connected to the outer walls of two sides of the elastic air bag 104, through holes are formed in the outer walls of two sides of the hollow ball shell 101, hole plugs 107 matched with the through holes are fixedly connected to one ends of the two linkage rods 106 far away from the elastic air bag 104, the two hole plugs 107 are respectively inserted into the two through holes, glucose solution 117 is filled in the chemotactic balls 103, a plurality of attaching fluff 118 are fixedly connected to the outer wall of the chemotactic balls 103, the chemotactic balls 103 are made of semipermeable membrane materials, membrane holes on the chemotactic balls 103 are smaller than glucose molecules, after the microorganism collecting balls are placed into a screening box 002, and viable microorganism strains are high-quality strains which can be properly used in soil environments after the microorganism collecting balls are screened, the microorganism strains can be attracted into the hollow ball 101, the high-quality microorganism collecting balls can be moved towards the hollow ball 101 and then the hollow ball 101.
Referring to fig. 2-4, a shaping net 105 is disposed on the inner wall of the elastic air bag 104, a discharge hole 108 is disposed at the top end of the hollow spherical shell 101, an air duct 109 is disposed in the middle of the discharge hole 108, the air duct 109 sequentially penetrates through the elastic air bag 104 and the outer wall of the top end of the shaping net 105 and extends into the shaping net 105, support rods 110 are fixedly connected to the outer walls of two sides of the air duct 109, one end of the support rod 110 away from the air duct 109 is fixedly connected with the inner wall of the hollow spherical shell 101, an elastic air-inflating bag 111 is disposed above the air duct 109, an air suction check valve 112 is disposed on the top end of the elastic air-inflating bag 111, an air outlet check valve 113 is disposed on the outer wall of one side of the air duct 109, one end of the air outlet check valve 113 away from the elastic air-inflating bag 111 is communicated with the air duct 109, through the arrangement of the elastic air bag 104, the linkage rod 106, the hole plug 107, the air duct 109, the elastic air pumping bag 111, the air suction one-way valve 112 and the air outlet one-way valve 113, after the microorganism collecting ball is put into the screening box 002, a proper amount of liquid culture medium is poured into the hollow ball cover 101 after repeatedly squeezing the elastic air pumping bag 111 to expand the hollow ball cover 105 until the hole plug 107 is moved towards the outside of the hollow ball cover 101, the through hole is conducted until the hole plug 107 is separated from the through hole, and then the high-quality strain can be moved towards the chemotactic ball 103, after collection, the air release valve 114 can be opened to prevent the air from the hollow ball cover 105, so that the hole plug 107 is plugged again, then the microorganism collecting ball is lifted up to control water, the moisture in the hollow ball cover 101 flows out through the water-controlling semipermeable membrane 102, then a proper amount of liquid culture medium is poured into the hollow ball cover 101, after the microorganism collecting ball is inverted, the high-quality strain can be poured out through the drain hole 108, thereby taking out the high-quality strain, the outer wall of the air duct 109 is fixedly sleeved with a guide hopper 115, so that the elastic air bag 111 can be prevented from being polluted when the high-quality strain is poured out, and the outer wall of the hollow spherical shell 101 is fixedly connected with two symmetrically arranged holding handles 116, so that the microorganism collecting balls can be conveniently lifted and inverted.
Referring to fig. 6, positioning blocks 201 matching with the hole plugs 107 are disposed on inner walls of two sides of the screening box 002, when the elastic air-beating bag 111 is pressed to move the hole plugs 107 to the outside of the hollow spherical shell 101, the elastic air-beating bag 111 is stopped from being pressed after the hole plugs 107 are moved to abut against the positioning blocks 201, on one hand, a positioning effect can be achieved, on the other hand, the microorganism collecting balls can be prevented from toppling over, after the microorganism collecting balls are placed in the screening box 002 in S5, the liquid level of the strain screening culture solution is not lower than the bottom end of the hole plugs 107, and high-quality strains cannot enter the hollow spherical shell 101 through the through holes due to the fact that the liquid level is too low.
The invention creatively increases the procedures of screening and culturing strains by utilizing the soil of a target area, firstly samples the target area before preparing the microbial inoculum, removes impurities from soil samples, sterilizes, adds water and mixes the soil samples to prepare screening liquid, screens the strains by the screening liquid to obtain high-quality strains which can survive in the screening liquid, namely the strains which can adapt to the soil environment of the target area, so that the survival rate of microorganisms in the microbial inoculum can be greatly improved when the prepared microbial inoculum is used in the target area, the effect of the microbial inoculum is effectively improved, the practicability is strong, and the microbial collection balls for collecting the high-quality strains are arranged, can quickly and effectively collect the screened high-quality strains based on chemotaxis of the microorganisms, and can effectively improve the preparation efficiency.
The above description is only of the preferred embodiments of the present invention; the scope of the invention is not limited in this respect. Any person skilled in the art, within the technical scope of the present disclosure, may apply to the present invention, and the technical solution and the improvement thereof are all covered by the protection scope of the present invention.
Claims (8)
1. A preparation method of a microbial agent for soil acidification areas is characterized by comprising the following steps: the method comprises the following steps:
s1, taking a proper amount of in-situ soil in a target area as a soil sample, removing impurities and garbage in the soil sample, and obtaining a impurity-removed soil sample;
s2, performing high-temperature sterilization on the impurity-removed soil sample to obtain a sterilized impurity-removed soil sample;
s3, fully mixing and stirring the sterilized impurity-removed soil sample and water, naturally precipitating, and taking supernatant as screening liquid;
s4, pouring the screening liquid into a screening box (002), then adding beneficial microorganism strains and a liquid culture medium into the screening box (002), mixing to form strain screening culture liquid, and carrying out directional screening on the strains for 2-3d;
s5, placing the microorganism collecting balls into a screening box (002), and collecting beneficial microorganism strains which survive in the screening box (002);
s6, taking out the microorganism strains collected by the microorganism collecting balls to obtain high-quality strains, and carrying out propagation on the high-quality strains;
s7, fermenting the high-quality strain subjected to propagation in the step S6 to obtain bacterial liquid;
s8, uniformly mixing the bacterial liquid in the step S7 with the fermentation material, and then drying in a drying oven to obtain the microbial agent;
the microbial collection ball comprises a hollow ball shell (101), a water control semipermeable membrane (102) is embedded and installed at the bottom end of the hollow ball shell (101), two symmetrically arranged chemotactic balls (103) are fixedly connected to the inner wall of the hollow ball shell (101), an elastic air bag (104) is arranged in the middle of the hollow ball shell (101), a linkage rod (106) is fixedly connected to the outer walls of the two sides of the elastic air bag (104), through holes are formed in the outer walls of the two sides of the hollow ball shell (101), hole plugs (107) matched with the through holes are fixedly connected to one ends, far away from the elastic air bag (104), of the two linkage rods (106), and the two hole plugs (107) are spliced with the two through holes respectively; the inner wall of elastic air bag (104) is provided with design net (105), discharge port (108) have been seted up on the top of hollow spherical shell (101), the middle part of discharge port (108) runs through and is provided with air duct (109), air duct (109) run through elastic air bag (104), the top outer wall of design net (105) in proper order and extend to in design net (105), all fixedly connected with bracing piece (110) on the outer wall of air duct (109) both sides, the one end that air duct (109) were kept away from to bracing piece (110) and the inner wall fixed connection of hollow spherical shell (101).
2. The method for preparing the microbial agent for soil acidification areas according to claim 1, which is characterized by comprising the following steps: the high-temperature sterilization temperature in the step S2 is 90-130 ℃, the sterilization time is 20-40min, and the mass ratio of the sterilization impurity-removing soil sample to the water in the step S3 is 1:1-2.
3. The method for preparing the microbial agent for soil acidification areas according to claim 1, which is characterized by comprising the following steps: the beneficial microorganism strain in the S4 is one or more of lactobacillus, rhizobium, bacillus and saccharomycetes, and the volume ratio of beneficial microorganism to culture medium to screening liquid is 1:20-30:50-100.
4. The method for preparing the microbial agent for soil acidification areas according to claim 1, which is characterized by comprising the following steps: the fermentation material in the step S8 is formed by uniformly mixing bean flour and bran according to the volume ratio of 1:4, and the volume ratio of the fermentation material to the bacterial liquid in the step S8 is 1:2-3.
5. The method for preparing the microbial agent for soil acidification areas according to claim 1, which is characterized by comprising the following steps: the air guide pipe is characterized in that an elastic air inflation bag (111) is arranged above the air guide pipe (109), an air suction one-way valve (112) is communicated with the top end of the elastic air inflation bag (111), an air outlet one-way valve (113) is communicated with the bottom end of the elastic air inflation bag (111), and one end, far away from the elastic air inflation bag (111), of the air outlet one-way valve (113) is communicated with the air guide pipe (109).
6. The method for preparing the microbial agent for soil acidification areas, which is characterized by comprising the following steps of: an air release valve (114) is arranged on the outer wall of one side of the air duct (109), a guide hopper (115) is fixedly sleeved on the outer wall of the air duct (109), and two holding handles (116) which are symmetrically arranged are fixedly connected on the outer wall of the hollow spherical shell (101).
7. The method for preparing the microbial agent for soil acidification areas, which is characterized by comprising the following steps of: the chemotactic ball (103) is filled with glucose solution (117), a plurality of attached villi (118) are fixedly connected to the outer wall of the chemotactic ball (103), the chemotactic ball (103) is made of semipermeable membrane materials, and membrane holes on the chemotactic ball (103) are smaller than glucose molecules.
8. The method for preparing the microbial agent for soil acidification areas according to claim 1, which is characterized by comprising the following steps: the inner walls of the two sides of the screening box (002) are provided with positioning blocks (201) matched with the hole plugs (107), and after the microorganism collecting balls are placed in the screening box (002) in S5, the liquid level of the strain screening culture solution is not lower than the bottom ends of the hole plugs (107).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110453802.8A CN113337419B (en) | 2021-04-26 | 2021-04-26 | Preparation method of microbial agent for soil acidification areas |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110453802.8A CN113337419B (en) | 2021-04-26 | 2021-04-26 | Preparation method of microbial agent for soil acidification areas |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113337419A CN113337419A (en) | 2021-09-03 |
| CN113337419B true CN113337419B (en) | 2023-06-13 |
Family
ID=77468659
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110453802.8A Active CN113337419B (en) | 2021-04-26 | 2021-04-26 | Preparation method of microbial agent for soil acidification areas |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113337419B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2845408A1 (en) * | 2012-11-30 | 2014-05-30 | Manas Ranjan Banerjee | Phosphate solubilizing rhizobacteria bacillus firmus as biofertilizer to increase canola yield |
| CN104830662A (en) * | 2014-02-10 | 2015-08-12 | 中国石油化工股份有限公司 | Device and method for quantitatively evaluating chemotaxis of bacteria to crude oil |
| WO2017180909A1 (en) * | 2016-04-13 | 2017-10-19 | Nextgen Jane, Inc. | Sample collection and preservation devices, systems and methods |
| CN108496616A (en) * | 2018-02-12 | 2018-09-07 | 暨南大学 | A method of promote agrobacterium to be colonized in plant rhizosphere |
| CN108715816A (en) * | 2018-04-12 | 2018-10-30 | 暨南大学 | A method of induction aimed strain forms chemotactic ring on semisolid tablet |
| CN109055277A (en) * | 2018-09-17 | 2018-12-21 | 安徽省交通航务工程有限公司 | A method of screening the resistance to heavy metal nitrogen-fixing bacteria of acidophilus from soil |
-
2021
- 2021-04-26 CN CN202110453802.8A patent/CN113337419B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2845408A1 (en) * | 2012-11-30 | 2014-05-30 | Manas Ranjan Banerjee | Phosphate solubilizing rhizobacteria bacillus firmus as biofertilizer to increase canola yield |
| CN104830662A (en) * | 2014-02-10 | 2015-08-12 | 中国石油化工股份有限公司 | Device and method for quantitatively evaluating chemotaxis of bacteria to crude oil |
| WO2017180909A1 (en) * | 2016-04-13 | 2017-10-19 | Nextgen Jane, Inc. | Sample collection and preservation devices, systems and methods |
| CN108496616A (en) * | 2018-02-12 | 2018-09-07 | 暨南大学 | A method of promote agrobacterium to be colonized in plant rhizosphere |
| CN108715816A (en) * | 2018-04-12 | 2018-10-30 | 暨南大学 | A method of induction aimed strain forms chemotactic ring on semisolid tablet |
| CN109055277A (en) * | 2018-09-17 | 2018-12-21 | 安徽省交通航务工程有限公司 | A method of screening the resistance to heavy metal nitrogen-fixing bacteria of acidophilus from soil |
Non-Patent Citations (3)
| Title |
|---|
| "A dodecapeptide (YQVTQSKVMSHR) exhibits antibacterial effect and induces cell aggregation in Escherichia coli";Lin, KC等;《APPLIED MICROBIOLOGY AND BIOTECHNOLOGY》;20120531;第94卷(第3期);第755-762页 * |
| "Integration of chemotaxis, transport and catabolism in Pseudomonas putida and identification of the aromatic acid chemoreceptor PcaY";Luu, RA等;《MOLECULAR MICROBIOLOGY》;20150430;第96卷(第1期);第134-147页 * |
| "表面活性剂与金属离子对采油微生物局部富集的影响";盖立学等;《南开大学学报(自然科学版)》;20101231;第43卷(第4期);第36-41页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113337419A (en) | 2021-09-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101100646A (en) | Paecilomyces lilacinus and application thereof | |
| CN109601248A (en) | A kind of field expanding propagation method of arbuscular mycorrhizal fungi | |
| CN107881132A (en) | A kind of the fixed nitrogen phosphorus decomposing Promoting bacteria that can be used for desert plant and its microbial inoculum are applied with it | |
| CN101558766B (en) | Trichoderma solid granules for preventing and controlling tobacco soil-borne fungus diseases and preparation method thereof | |
| CN113337419B (en) | Preparation method of microbial agent for soil acidification areas | |
| CN105850535B (en) | A method of it improving M. truncatula salt resistance and does harm to ability | |
| CN105483020B (en) | Application of the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 in prevention and treatment notoginseng root rot | |
| CN106434490A (en) | Ginseng bacterium TY15-2 with effects of disease prevention and growth promotion and application thereof | |
| CN104651286B (en) | A kind of soybean nodulation endophyte DEnt1302 and its preparation method and application, microbial inoculum and preparation method thereof | |
| CN107541468B (en) | Trichoderma brachiatum, microbial inoculum, method and application of trichoderma brachiatum in degradation of imazethapyr | |
| CN103392744A (en) | Trichoderma preparation for controlling pepper phytophthora blight and field tank mix pesticide containing the same | |
| CN103026902B (en) | Method for germinating ganoderma spores | |
| CN205993154U (en) | Mycorrhizal fungi locellus culture apparatus | |
| CN107036873B (en) | Device and method for in-situ extraction of arbuscular mycorrhizal plant root secretions | |
| CN105483041B (en) | Application of the sophora tonkinensis Gapnep endogenetic bacteria B22 in prevention and treatment Alternaria panax | |
| CN105462847B (en) | Application of the sophora tonkinensis Gapnep endogenetic fungus TRXY-46 in prevention and treatment notoginseng root rot | |
| CN107517833B (en) | Rhizoma Gastrodiae pollen planting bag fills method for culturing seedlings | |
| CN105462895B (en) | Application of the sophora tonkinensis Gapnep endogenetic bacteria B22 in prevention and treatment Radix Notoginseng anthracnose | |
| CN110560469A (en) | Method for restoring uranium-cadmium combined polluted soil by using uranium-cadmium-resistant fungi enhanced plants | |
| CN105462897B (en) | Application of the sophora tonkinensis Gapnep endogenetic bacteria B21 in prevention and treatment Radix Notoginseng anthracnose | |
| CN108949582A (en) | A kind of high cadmium resistance Eight Strains of Pleurotus Ostreatus and its application | |
| CN105462891B (en) | Application of the sophora tonkinensis Gapnep endogenetic bacteria B21 in prevention and treatment notoginseng root rot | |
| CN105462896B (en) | Application of the sophora tonkinensis Gapnep endogenetic bacteria B29 in prevention and treatment Alternaria panax | |
| CN107828690A (en) | The plant phosphorus decomposing fixed nitrogen Promoting bacteria and its microbial inoculum that stony desert area can be used for are applied with it | |
| CN104593302B (en) | Genuine Radix Astragali nitrogen-fixing bacteria new strains t16 and zymotic fluid preparation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20230518 Address after: 110400 Gaosanjiazi Village, Xiushui Hezi Town, Faku County, Shenyang City, Liaoning Province Applicant after: SHENYANG HUAQINGYUAN AGRICULTURAL DEVELOPMENT Co.,Ltd. Address before: 332000 No. 71, Xiehe Avenue, Wuning County, Jiujiang City, Jiangxi Province Applicant before: Li Xuechen |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |