CN106047771A - Aerobic bacterium-anaerobic bacterium composite microbe liquid inocula and preparation method thereof - Google Patents
Aerobic bacterium-anaerobic bacterium composite microbe liquid inocula and preparation method thereof Download PDFInfo
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Abstract
The invention discloses aerobic bacterium-anaerobic bacterium composite microbe liquid inocula and a preparation method thereof. The composite microbe liquid inocula comprise six liquid aerobic microbial agents and four liquid anaerobic microbial agents. The liquid aerobic microbial agents comprise a bacillus megaterium agent, a liquid Azotobacter chroococcum agent, a liquid bacillus mucilaginosus agent, a liquid streptomyces microflavus agent, a liquid bacillus subtilis agent and a liquid bacillus licheniformis agent. The liquid anaerobic microbial agents comprise liquid photosynthetic bacteria, a liquid lactobacillus plantarum agent, a liquid lactobacillus acidophilus agent and a liquid lactobacillus delbrueckii agent. The preparation method comprises culturing six types of aerobic bacteria to obtain six liquid aerobic microbial agents, culturing four types of anaerobic bacteria to obtain four mixed liquid anaerobic microbial agents, and injecting the six liquid aerobic microbial agents into the mixed liquid anaerobic microbial agents to obtain the composite microbe liquid inocula. The composite microbe liquid inocula have a quality guarantee period for one year or more at the normal temperature and have functions of nitrogen fixation, phosphate dissolution, potassium release, disease prevention and soil improvement.
Description
Technical field
The present invention relates to belong to agricultural microorganism, ecologic planting and microbial bacterial agent technical field, it is a kind of good to particularly relate to
Microbial liquid microbial inoculum that oxygen bacterium is compound with anaerobe and preparation method thereof.
Background technology
China uses chemical fertilizer due to long-term excess, has had resulted in substantial amounts of soil compaction and acidifying, the permeability of soil,
Oxygen delivery capacity and maintain nutrient balance ability to reduce, in soil, beneficial microbe drastically reduces, the residual of harmful substance and harmful
Microbial growth, controls root system of plant to nutrient, the absorption of moisture, causes that plant growth is bad, quality declines, various diseases
It is on the rise.The increase of crops internal chemistry of pesticide residual, seriously threatens the health and safety of the mankind simultaneously;Chemical fertilizer mistake
Amount uses the pollution of area source caused, and also exacerbates the heavy damage of ecological environment.Recently, National agricultural portion has been proposed for chemical fertilizer
The action scheme of pesticide zero growth rate.Agricultural product increasing both production and income is ensured again, it is achieved supply while fertilizer and pesticide zero growth rate to be realized
To the effective supply of side, it is necessary to can the product of effective replacing fertilizer pesticide.It is possible to replacing fertilizer and reduce pesticide
Microbial manure becomes the urgent need of the current development ecological agriculture, improves Soil structure with microorganism formulation, Crop Improvement quality,
Realize increasing both production and income, it has also become the inexorable trend of modern agriculture, be the inevitable choice developing green sustainable Efficient Eco-agriculture.
The most various microbial manures emerge in multitude like the mushrooms after rain, but fertilizer efficiency substantially can meet or exceed chemical fertilizer
Product the most actually rare, its main cause: is that microorganism fungus kind is single, can't resolve the problems of plant growth;Two
Being that preservative time is short, fertilizer efficiency is inconspicuous, and the most single microbial inoculum or composite bacteria agent capable are all difficult to solve liquid bacterial agent corruption, have
The problem that effect bacterium is dead, storage life is short.
Summary of the invention
It is an object of the invention to provide a kind of aerobic bacteria microbial liquid microbial inoculum compound with anaerobe and preparation method thereof,
This microbial liquid microbial inoculum and preparation method thereof solves that microbial bacterial agent strain is single, storage life is short and fertilizer efficiency is unconspicuous
Problem.
To achieve the above object of the invention, the microbial liquid microbial inoculum that a kind of aerobic bacteria that the present invention uses is combined with anaerobe
Technical scheme be: described microbial liquid microbial inoculum includes that 6 kinds of aerobic microbial inoculums of liquid cultivated with fluid medium and 4 kinds are with liquid
The liquid anaerobic microbial inoculum of body culture medium culturing, the aerobic microbial inoculum of described liquid is 3:7 ~ 7:3 with the mass ratio of liquid anaerobic microbial inoculum;Institute
State the aerobic microbial inoculum of liquid to be made up of the microbial inoculum of following mass parts:
Liquid bacillus megaterium agent 1 ~ 3 mass parts,
Liquid blown-ball Azotobacter agent 1 ~ 3 mass parts,
Liquid colloid bacillus cereus agent 1 ~ 3 mass parts,
Liquid streptomyces microflavus agent 1 mass parts,
Liquid bacillus subtilis microbial inoculum 1 ~ 2 mass parts,
Liquid Bacillus licheniformis agent 1 ~ 2 mass parts;
Described liquid anaerobic microbial inoculum is made up of the microbial inoculum of following mass parts:
Liquid photosynthetic microbial inoculum 1 ~ 2 mass parts,
Liquid plant breast microbial inoculum 1 ~ 2 mass parts,
Liquid bacillus acidophilus's agent 1 ~ 5 mass parts,
Liquid Deshi Lactobacillus agent 1 ~ 5 mass parts;
Wherein, the good bacteria content in the aerobic microbial inoculum of each liquid is more than or equal to 200,000,000/milliliter, each liquid anaerobic bacterium
Anaerobe content in agent is more than or equal to 200,000,000/milliliter.
Relevant content in technique scheme is explained as follows:
1, in such scheme, described microbial liquid microbial inoculum also includes trace element, and the addition of trace element is described micro-life
The 0.1 ~ 0.2% of thing liquid bacterial agent quality, described trace element in B, Zn, Mn, Cu, Co any one or at least two with
On mixture.
To achieve the above object of the invention, the microbial liquid microbial inoculum that a kind of aerobic bacteria that the present invention uses is combined with anaerobe
The technical scheme of preparation method be: described preparation method comprises the following steps:
6 kinds of aerobic bacterias are cultivated and are made 6 kinds of aerobic microbial inoculums of liquid by the first step respectively,
(1) prepare respectively bacillus megaterium, blown-ball Azotobacter, colloid bacillus cereus, streptomyces microflavus, bacillus subtilis with
And the liquid aerobic bacteria culture medium of Bacillus licheniformis, more each self-corresponding to 6 kinds of liquid aerobic bacteria inoculation of mediums respectively
Strain, at 30 ~ 37 DEG C, carries out the one-level amplification culture of strain under the conditions of 120 ~ 140r/min, obtain 6 kinds of aerobic bacterium solution of one-level;
(2) 6 kinds of aerobic bacterium solution of one-level are taken the most respectively to 6 kinds of described each self-corresponding strains of liquid aerobic bacteria inoculation of medium,
30 ~ 37 DEG C, carry out two grades of amplification culture of strain under the conditions of 120 ~ 140r/min, obtain 6 kinds of two grades of aerobic bacterium solution, wherein,
The inoculum concentration of every kind of aerobic bacterium solution of one-level is the 0.9 ~ 1.1% of its liquid aerobic bacteria culture medium quality;
(3) 6 kinds of two grades of aerobic bacterium solution are taken again to 6 kinds of described each self-corresponding strains of liquid aerobic bacteria inoculation of medium, 30 ~
37 DEG C, carry out three grades of amplification culture of strain under the conditions of 120 ~ 140r/min, obtain 6 kinds of aerobic microbial inoculums of liquid, wherein, every kind two
Inoculum concentration is its liquid aerobic bacteria culture medium quality the 1 ~ 1.5% of the aerobic bacterium solution of level;
The liquid anaerobic microbial inoculum of 4 kinds of mixing is made in 4 kinds of anaerobic culturees by second step,
(1) the liquid anaerobic bacterium culture medium of photosynthetic bacteria, Lactobacillus plantarum, bacillus acidophilus and Deshi Lactobacillus is prepared respectively,
In 4 kinds of liquid anaerobic bacterium culture mediums, inoculate each self-corresponding strain the most respectively, stand under the conditions of 30 ~ 37 DEG C, carry out strain
One-level amplification culture, obtain 4 kinds of one-level anaerobism bacterium solution;
(2) take 4 kinds of one-level anaerobism bacterium solution the most respectively in 4 kinds of described liquid anaerobic bacterium culture mediums, inoculate each self-corresponding strain,
Stand under the conditions of 30 ~ 37 DEG C, carry out two grades of amplification culture of strain, obtain 4 kinds of secondary anaerobic bacterium solution, wherein, every kind of one-level
The inoculum concentration of anaerobism bacterium solution is the 1 ~ 1.5% of its liquid anaerobic bacterium culture medium quality;
(3) preparation mixed fermentation fluid medium, takes described 4 kinds of secondary anaerobic bacterium solution in described mixed fermentation fluid medium
Inoculation mixing, 30 ~ 37 DEG C, stand mixed culture under the conditions of illumination 2800 ~ 3200Ix, obtain the liquid anaerobic microbial inoculum of mixing;
6 kinds of aerobic microbial inoculums of liquid are slowly injected into the liquid of described mixing by the 3rd step by the formula proportion of a upper technical scheme
In body anaerobism microbial inoculum and stir, stand, behind isotropic stable, obtain described microbial liquid microbial inoculum.
Relevant content in technique scheme is explained as follows:
1, in such scheme, in described 3rd step, being simultaneously introduced trace element, the addition of trace element is described microorganism
The 0.1 ~ 0.2% of liquid bacterial agent quality, described trace element is selected from B, Zn, Mn, Cu, Co more than any one or at least two
Mixture.
2, in such scheme, the liquid aerobic bacteria culture medium prescription of described bacillus megaterium is according to the matter shared by each component
Amount percentage composition is composed of the following components: sucrose 1%, dipotassium hydrogen phosphate 0.1%, ammonium sulfate 0.2%, soluble starch 2%, calcium carbonate
0.5%, Oleum Glycines 0.5%, sterilized water 95.7%;
The liquid aerobic bacteria culture medium prescription of described blown-ball Azotobacter according to the weight/mass percentage composition shared by each component by following group
It is grouped into: sucrose 1%, dipotassium hydrogen phosphate 0.1%, yeast extract 0.5%, magnesium sulfate 0.05%, calcium carbonate 0.2%, sodium molybdate 0.001%,
Borax 0.001%, ferrous sulfate 0.001%, sodium chloride 0.002%, sterilized water 98.145%;
The liquid aerobic bacteria culture medium prescription of described colloid bacillus cereus according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 2%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.04%, yeast extract 0.2%, calcium carbonate 0.2%, ferrous sulfate
0.001%, sodium chloride 0.04%, sterilized water 97.419%;
The liquid aerobic bacteria culture medium prescription of described streptomyces microflavus according to the weight/mass percentage composition shared by each component by following group
It is grouped into: soluble starch 2%, dipotassium hydrogen phosphate 0.05%, potassium nitrate 0.1%, magnesium sulfate 0.05%, sodium chloride 0.005%, sulphuric acid
Ferrous 0.001%, sterilized water 97.794%;
The liquid aerobic bacteria culture medium prescription of described bacillus subtilis according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 1%, dipotassium hydrogen phosphate 0.05%, yeast extract 0.05%, Carnis Bovis seu Bubali cream 0.02%, peptone 0.05%, calcium carbonate
0.025%, sodium chloride 0.02%, manganese sulfate 0.001%, sterilized water 98.784%;
The liquid aerobic bacteria culture medium prescription of described Bacillus licheniformis according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 1%, dipotassium hydrogen phosphate 0.05%, yeast extract 0.05%, Carnis Bovis seu Bubali cream 0.02%, peptone 0.05%, calcium carbonate
0.025%, sodium chloride 0.02%, manganese sulfate 0.001%, sterilized water 98.784%;
The liquid anaerobic bacterium culture medium formula of described photosynthetic bacteria is: the 3.3g Han sodium acetate, ammonium chloride 0.6g in every 1 liter of sterilized water,
Magnesium sulfate 0.5g, yeast extract 1.5g;
The liquid anaerobic bacterium culture medium formula of described Lactobacillus plantarum is: the 20g Han sucrose in every 1 liter of sterilized water, yeast extract
25g, dipotassium hydrogen phosphate 2g;
The liquid anaerobic bacterium culture medium formula of described bacillus acidophilus is: the 5g Han sodium acetate in every 1 liter of sterilized water, ammonium citrate
2g, magnesium sulfate 0.5g, yeast extract 6g, Carnis Bovis seu Bubali cream 12g, dipotassium hydrogen phosphate 2g, peptone 10g, manganese sulfate 0.05g;
The liquid anaerobic bacterium culture medium formula of described Deshi Lactobacillus is: the 50g Han whey powder in every 1 liter of sterilized water, calcium carbonate
2g, yeast extract 10g;
Described mixed fermentation liquid culture based formulas is composed of the following components according to the weight/mass percentage composition shared by each component: sucrose
4%, yeast extract 0.4%, dipotassium hydrogen phosphate 0.2%, sterilized water 95.4%.
The design feature of the present invention is: the microbial liquid microbial inoculum of the present invention is that aerobic flora is composited with anaerobic bacteria flora
Microbial liquid microbial inoculum, its feature for preparing A and B two class complex micro organism fungicide the most respectively, A microbial inoculum by 6 kinds of aerobic bacterias and
Facultative aerobe forms, and every kind of aerobic bacteria uses aerobic fermentation process individually to ferment.B microbial inoculum is by 4 kinds of anaerobe and amphimicrobian
Bacterium forms, and 4 kinds of anaerobe use anaerobic ferment process to inoculate respectively, carry out first class inoculum cultivation, detest 4 kinds the most in proportion
Oxygen bacterium mixes, and mixed strain carries out second class inoculum mixed culture, obtains compound B preparation.Last again by 6 kinds of A microbial inoculums difference
It is mixed in proportion in B microbial inoculum.This feature is advantageous in that, in the aerobic fermentation stage to 6 kinds of bacterial strain single culture, can ensure bacterium
Kind of purity and functional on the premise of, promote that aerobic bacteria quickly increases breeding.After aerobic fermentation completes, when 6 kinds of liquid aerobic bacterias
After agent is mixed in the B microbial inoculum of anaerobic fermentation in proportion, aerobic flora is suppressed by anaerobic bacteria flora and mild acid conditions, increases numerous
Growing and become to slow down, most bacterial strains define spore, form the environmental condition of the most excellent symbiosis of antagonism, often between bacterial strain
The lower liquid bacterial agent of temperature can preserve the long period.
The present invention compared with prior art, has the advantages that
(1) conventional microbial bacterial agent is under the conditions of liquid, and bacterial strain several minutes to a few hours just breeding one generation, due to the fastest
Speed flourish, bacterial strain will soon deformity dead, microbial inoculum will blackening smelly, after dilution, most floras are for this microbial inoculum
Weak through becoming deformity, it is impossible to playing its effective efficiency, under normal temperature condition, storage life is the shortest, the most only the several months.And this
In bright microbial liquid microbial inoculum, living bacteria count is many, long shelf-life, and two kinds of microbial inoculums of A, B ferment the most multiple respectively
Joining, after compounding in the environment of faintly acid, the growth conditions of flora is inhibited, and nutrient and metabolizable energy have been got back mutually simultaneously
Mend, between flora, define the excellent symbiotic environment of interdependent symbiosis, the most not antagonism.Thus, the growth of flora is inhibited, and exists equally
Under conditions of liquid room temperature, flora becomes slowly to grow, and most bacterial strains form spore states, and bacterial strain vigor is maintained and temporarily
In a dormant state.Under normal temperature condition, the microbial liquid microbial inoculum preservation effect duration of the present invention can reach more than 1 year, dilute
After releasing, bacterial strain again can quickly restoration ecosystem playing a role.
(2) multi-cultur es Synergistic in the microbial liquid microbial inoculum of the present invention, complete function.The present invention uses A(that is 6 kind
Aerobic flora), B(that is 4 kind anaerobic bacteria flora) two kinds of microbial inoculums ferment method the most compounding respectively, can make more
Difference in functionality bacterial strain compounds symbiosis.Use rear fertilizer efficiency obvious, fixed nitrogen, phosphorus decomposing, potassium decomposing, antibacterial, pest-resistant, promotion life can not only be played
Long effect, additionally it is possible to improve Soil structure, prevent and treat soil-borne disease;Efficient growth, the prominent improvement of quality can be realized for crop, be
Green, organic, ecologic planting creates favorable conditions.
Detailed description of the invention
Below in conjunction with embodiment, the invention will be further described:
Embodiment 1: microbial liquid microbial inoculum that a kind of aerobic bacteria is compound with anaerobe and preparation method thereof
Described microbial liquid microbial inoculum includes that 6 kinds of aerobic microbial inoculums of liquid cultivated with fluid medium and 4 kinds are with fluid medium
The liquid anaerobic microbial inoculum cultivated, the aerobic microbial inoculum of described liquid is 9:8 with the mass ratio of liquid anaerobic microbial inoculum;Described liquid aerobic bacteria
Agent is made up of the microbial inoculum of following mass parts:
Liquid bacillus megaterium agent 2 mass parts,
Liquid blown-ball Azotobacter agent 2 mass parts,
Liquid colloid bacillus cereus agent 2 mass parts,
Liquid streptomyces microflavus agent 1 mass parts,
Liquid bacillus subtilis microbial inoculum 1 mass parts,
Liquid Bacillus licheniformis agent 1 mass parts;
Described liquid anaerobic microbial inoculum is made up of the microbial inoculum of following mass parts:
Liquid photosynthetic microbial inoculum 1 mass parts,
Liquid plant breast microbial inoculum 1 mass parts,
Liquid bacillus acidophilus's agent 3 mass parts,
Liquid Deshi Lactobacillus agent 3 mass parts;
Wherein, the good bacteria content in the aerobic microbial inoculum of each liquid is more than or equal to 200,000,000/milliliter, each liquid anaerobic bacterium
Anaerobe content in agent is more than or equal to 200,000,000/milliliter.
Described microbial liquid microbial inoculum also includes trace element, and the addition of trace element is described microbial liquid microbial inoculum
The 0.1% of quality, described trace element is selected from the mixture of B, Zn, Mn three.
Preparation method comprises the following steps:
6 kinds of aerobic bacterias are cultivated and are made 6 kinds of aerobic microbial inoculums of liquid by the first step respectively,
(1) prepare respectively bacillus megaterium, blown-ball Azotobacter, colloid bacillus cereus, streptomyces microflavus, bacillus subtilis with
And the liquid aerobic bacteria culture medium of Bacillus licheniformis, more each self-corresponding to 6 kinds of liquid aerobic bacteria inoculation of mediums respectively
Strain, at 30 DEG C, 120r/min, pH value is cultivated 48h under the conditions of 6.5 ~ 7.5, is carried out the one-level amplification culture of strain, obtain 6
Plant the aerobic bacterium solution of one-level;
(2) 6 kinds of aerobic bacterium solution of one-level are taken the most respectively to 6 kinds of described each self-corresponding strains of liquid aerobic bacteria inoculation of medium,
30 DEG C, cultivate 48h under the conditions of 120r/min and carry out two grades of amplification culture of strain, obtain 6 kinds of two grades of aerobic bacterium solution, wherein,
The inoculum concentration of every kind of aerobic bacterium solution of one-level is the 1% of its liquid aerobic bacteria culture medium quality;
(3) 6 kinds of two grades of aerobic bacterium solution are taken again to 6 kinds of described each self-corresponding strains of liquid aerobic bacteria inoculation of medium, 30
DEG C, 120r/min, pH value under the conditions of 6.5 ~ 7, cultivate 72h carry out three grades of amplification culture of strain, obtain 6 kinds of liquid aerobic bacterias
Agent, wherein, the inoculum concentration of every kind of two grades of aerobic bacterium solution is the 1% of its liquid aerobic bacteria culture medium quality;
The liquid anaerobic microbial inoculum of 4 kinds of mixing is made in 4 kinds of anaerobic culturees by second step,
(1) the liquid anaerobic bacterium culture medium of photosynthetic bacteria, Lactobacillus plantarum, bacillus acidophilus and Deshi Lactobacillus is prepared respectively,
In 4 kinds of liquid anaerobic bacterium culture mediums, inoculate each self-corresponding strain, quiescent culture 3d under the conditions of 30 DEG C the most respectively, carry out bacterium
The one-level amplification culture planted, obtains 4 kinds of one-level anaerobism bacterium solution;
(2) take 4 kinds of one-level anaerobism bacterium solution the most respectively in 4 kinds of described liquid anaerobic bacterium culture mediums, inoculate each self-corresponding strain,
Quiescent culture 6d under the conditions of 30 DEG C, carries out two grades of amplification culture of strain, obtains 4 kinds of secondary anaerobic bacterium solution, wherein, every kind one
Inoculum concentration is its liquid anaerobic bacterium culture medium quality the 1% of level anaerobism bacterium solution;
(3) preparation mixed fermentation fluid medium, the formula proportion according to claim 1 takes described 4 kinds of secondary anaerobic bacterium solution to institute
State in mixed fermentation fluid medium inoculation mixing, 30 DEG C, stand mixed culture 6d under the conditions of illumination 3000Ix, mixed
Liquid anaerobic microbial inoculum, wherein, total inoculum concentration of 4 kinds of secondary anaerobic bacterium solution is described mixed fermentation fluid medium quality
1%;
6 kinds of aerobic microbial inoculums of liquid are slowly injected in the liquid anaerobic microbial inoculum of described mixing by the 3rd step by above-mentioned formula proportion
And stir, stand, behind isotropic stable, obtain described microbial liquid microbial inoculum.
In described 3rd step, being simultaneously introduced trace element, the addition of trace element is described microbial liquid microbial inoculum
The 0.1% of quality, described trace element is the mixture of B, Zn, Mn three.
The liquid aerobic bacteria culture medium prescription of described bacillus megaterium according to the weight/mass percentage composition shared by each component by
Following components forms: sucrose 1%, dipotassium hydrogen phosphate 0.1%, ammonium sulfate 0.2%, soluble starch 2%, calcium carbonate 0.5%, Oleum Glycines
0.5%, sterilized water 95.7%;
The liquid aerobic bacteria culture medium prescription of described blown-ball Azotobacter according to the weight/mass percentage composition shared by each component by following group
It is grouped into: sucrose 1%, dipotassium hydrogen phosphate 0.1%, yeast extract 0.5%, magnesium sulfate 0.05%, calcium carbonate 0.2%, sodium molybdate 0.001%,
Borax 0.001%, ferrous sulfate 0.001%, sodium chloride 0.002%, sterilized water 98.145%;
The liquid aerobic bacteria culture medium prescription of described colloid bacillus cereus according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 2%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.04%, yeast extract 0.2%, calcium carbonate 0.2%, ferrous sulfate
0.001%, sodium chloride 0.04%, sterilized water 97.419%;
The liquid aerobic bacteria culture medium prescription of described streptomyces microflavus according to the weight/mass percentage composition shared by each component by following group
It is grouped into: soluble starch 2%, dipotassium hydrogen phosphate 0.05%, potassium nitrate 0.1%, magnesium sulfate 0.05%, sodium chloride 0.005%, sulphuric acid
Ferrous 0.001%, sterilized water 97.794%;
The liquid aerobic bacteria culture medium prescription of described bacillus subtilis according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 1%, dipotassium hydrogen phosphate 0.05%, yeast extract 0.05%, Carnis Bovis seu Bubali cream 0.02%, peptone 0.05%, calcium carbonate
0.025%, sodium chloride 0.02%, manganese sulfate 0.001%, sterilized water 98.784%;
The liquid aerobic bacteria culture medium prescription of described Bacillus licheniformis according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 1%, dipotassium hydrogen phosphate 0.05%, yeast extract 0.05%, Carnis Bovis seu Bubali cream 0.02%, peptone 0.05%, calcium carbonate
0.025%, sodium chloride 0.02%, manganese sulfate 0.001%, sterilized water 98.784%;
The liquid anaerobic bacterium culture medium formula of described photosynthetic bacteria is: the 3.3g Han sodium acetate, ammonium chloride 0.6g in every 1 liter of sterilized water,
Magnesium sulfate 0.5g, yeast extract 1.5g;
The liquid anaerobic bacterium culture medium formula of described Lactobacillus plantarum is: the 20g Han sucrose in every 1 liter of sterilized water, yeast extract
25g, dipotassium hydrogen phosphate 2g;
The liquid anaerobic bacterium culture medium formula of described bacillus acidophilus is: the 5g Han sodium acetate in every 1 liter of sterilized water, ammonium citrate
2g, magnesium sulfate 0.5g, yeast extract 6g, Carnis Bovis seu Bubali cream 12g, dipotassium hydrogen phosphate 2g, peptone 10g, manganese sulfate 0.05g;
The liquid anaerobic bacterium culture medium formula of described Deshi Lactobacillus is: the 50g Han whey powder in every 1 liter of sterilized water, calcium carbonate
2g, yeast extract 10g;
Described mixed fermentation liquid culture based formulas is composed of the following components according to the weight/mass percentage composition shared by each component: sucrose
4%, yeast extract 0.4%, dipotassium hydrogen phosphate 0.2%, sterilized water 95.4%.
The microbial liquid microbial inoculum of embodiment 1 contrast experiment in Rice Cropping:
Sample plot: 150 mu of experiment field, Donggou township, Wuxuan County, Guangxi.
Performance: experiment is carried out under the natural conditions of field, using seed rice is fragrant No. 1 of silk, and rice transplanting is planted, July time
The middle ten days, for the late rice of double cropping of rice.Experiment sets A, B, C3 and processes community, 50 mu, each community.Paddy field, community is connected, and soil is basic
Identical.The deerization board Chemical Mixed Fertilizer 50kg/ mu that Guangxi Luzhai chemical fertilizer factory produces all used by 3 community base manure, and nutrient content is 45%
(NPK3x15).All having done 2 Liquid Fertilizers in rice plant of tillering stage and 3 communities in boot stage to spray, wherein A community sprays embodiment 1
Microbial liquid microbial inoculum, each 100lm(this be what unit) dilute 150 times and spray 1 mu;B community sprays commercially available microorganism
Liquid bacterial agent, each 100lm dilutes 150 times and sprays 1 mu;C community sprays 5% dipotassium hydrogen phosphate solution 15kg.
Experimental result: during harvesting, B, C community spike of rice the same with the rice field of surrounding and rice straw all turn yellow, and A community spike of rice
Golden yellow rice straw still green, occur in that green bar maturation phenomenon.1180 jin, every mu of gulch yield A community, 960 jin, B community, C is little
920 jin of district.A community increases production 22.9% and 28.2% respectively than BC community.
Embodiment 2: microbial liquid microbial inoculum that a kind of aerobic bacteria is compound with anaerobe and preparation method thereof
Described microbial liquid microbial inoculum includes that 6 kinds of aerobic microbial inoculums of liquid cultivated with fluid medium and 4 kinds are with fluid medium
The liquid anaerobic microbial inoculum cultivated, the aerobic microbial inoculum of described liquid is 1:1 with the mass ratio of liquid anaerobic microbial inoculum;Described liquid aerobic bacteria
Agent is made up of the microbial inoculum of following mass parts:
Liquid bacillus megaterium agent 3 mass parts,
Liquid blown-ball Azotobacter agent 3 mass parts,
Liquid colloid bacillus cereus agent 3 mass parts,
Liquid streptomyces microflavus agent 1 mass parts,
Liquid bacillus subtilis microbial inoculum 2 mass parts,
Liquid Bacillus licheniformis agent 2 mass parts;
Described liquid anaerobic microbial inoculum is made up of the microbial inoculum of following mass parts:
Liquid photosynthetic microbial inoculum 2 mass parts,
Liquid plant breast microbial inoculum 2 mass parts,
Liquid bacillus acidophilus's agent 5 mass parts,
Liquid Deshi Lactobacillus agent 5 mass parts;
Wherein, the good bacteria content in the aerobic microbial inoculum of each liquid is more than or equal to 200,000,000/milliliter, each liquid anaerobic bacterium
Anaerobe content in agent is more than or equal to 200,000,000/milliliter.
Described microbial liquid microbial inoculum also includes trace element, and the addition of trace element is described microbial liquid microbial inoculum
The 0.1% of quality, described trace element is selected from the mixture of B, Cu, Co three.
Preparation method comprises the following steps:
6 kinds of aerobic bacterias are cultivated and are made 6 kinds of aerobic microbial inoculums of liquid by the first step respectively,
(1) prepare respectively bacillus megaterium, blown-ball Azotobacter, colloid bacillus cereus, streptomyces microflavus, bacillus subtilis with
And the liquid aerobic bacteria culture medium of Bacillus licheniformis, more each self-corresponding to 6 kinds of liquid aerobic bacteria inoculation of mediums respectively
Strain, at 37 DEG C, 140r/min, pH value is cultivated 48h under the conditions of 6.5 ~ 7.5, is carried out the one-level amplification culture of strain, obtain 6
Plant the aerobic bacterium solution of one-level;
(2) 6 kinds of aerobic bacterium solution of one-level are taken the most respectively to 6 kinds of described each self-corresponding strains of liquid aerobic bacteria inoculation of medium,
37 DEG C, cultivate 48h under the conditions of 140r/min and carry out two grades of amplification culture of strain, obtain 6 kinds of two grades of aerobic bacterium solution, wherein,
The inoculum concentration of every kind of aerobic bacterium solution of one-level is the 1.1% of its liquid aerobic bacteria culture medium quality;
(3) 6 kinds of two grades of aerobic bacterium solution are taken again to 6 kinds of described each self-corresponding strains of liquid aerobic bacteria inoculation of medium, 30
DEG C, 120r/min, pH value under the conditions of 6.5 ~ 7, cultivate 72h carry out three grades of amplification culture of strain, obtain 6 kinds of liquid aerobic bacterias
Agent, wherein, the inoculum concentration of every kind of two grades of aerobic bacterium solution is the 1.5% of its liquid aerobic bacteria culture medium quality;
The liquid anaerobic microbial inoculum of 4 kinds of mixing is made in 4 kinds of anaerobic culturees by second step,
(1) the liquid anaerobic bacterium culture medium of photosynthetic bacteria, Lactobacillus plantarum, bacillus acidophilus and Deshi Lactobacillus is prepared respectively,
In 4 kinds of liquid anaerobic bacterium culture mediums, inoculate each self-corresponding strain, quiescent culture 3d under the conditions of 37 DEG C the most respectively, carry out bacterium
The one-level amplification culture planted, obtains 4 kinds of one-level anaerobism bacterium solution;
(2) take 4 kinds of one-level anaerobism bacterium solution the most respectively in 4 kinds of described liquid anaerobic bacterium culture mediums, inoculate each self-corresponding strain,
Quiescent culture 6d under the conditions of 37 DEG C, carries out two grades of amplification culture of strain, obtains 4 kinds of secondary anaerobic bacterium solution, wherein, every kind one
Inoculum concentration is its liquid anaerobic bacterium culture medium quality the 1.5% of level anaerobism bacterium solution;
(3) preparation mixed fermentation fluid medium, takes described 4 kinds of secondary anaerobic bacterium solution by above-mentioned formula proportion and sends out to described mixing
Inoculation mixing in ferment fluid medium, 37 DEG C, stand mixed culture 6d under the conditions of illumination 3200Ix, the liquid obtaining mixing is detested
Oxygen microbial inoculum, wherein, total inoculum concentration of 4 kinds of secondary anaerobic bacterium solution is the 1.5% of described mixed fermentation fluid medium quality;
3rd step, is slowly injected into the liquid anaerobic of described mixing by 6 kinds of aerobic microbial inoculums of liquid formula proportion according to claim 1
In microbial inoculum and stir, stand, behind isotropic stable, obtain described microbial liquid microbial inoculum.
In described 3rd step, being simultaneously introduced trace element, the addition of trace element is described microbial liquid microbial inoculum
The 0.2% of quality, described trace element is the mixture of B, Cu, Co three.
The liquid aerobic bacteria culture medium prescription of described bacillus megaterium according to the weight/mass percentage composition shared by each component by
Following components forms: sucrose 1%, dipotassium hydrogen phosphate 0.1%, ammonium sulfate 0.2%, soluble starch 2%, calcium carbonate 0.5%, Oleum Glycines
0.5%, sterilized water 95.7%;
The liquid aerobic bacteria culture medium prescription of described blown-ball Azotobacter according to the weight/mass percentage composition shared by each component by following group
It is grouped into: sucrose 1%, dipotassium hydrogen phosphate 0.1%, yeast extract 0.5%, magnesium sulfate 0.05%, calcium carbonate 0.2%, sodium molybdate 0.001%,
Borax 0.001%, ferrous sulfate 0.001%, sodium chloride 0.002%, sterilized water 98.145%;
The liquid aerobic bacteria culture medium prescription of described colloid bacillus cereus according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 2%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.04%, yeast extract 0.2%, calcium carbonate 0.2%, ferrous sulfate
0.001%, sodium chloride 0.04%, sterilized water 97.419%;
The liquid aerobic bacteria culture medium prescription of described streptomyces microflavus according to the weight/mass percentage composition shared by each component by following group
It is grouped into: soluble starch 2%, dipotassium hydrogen phosphate 0.05%, potassium nitrate 0.1%, magnesium sulfate 0.05%, sodium chloride 0.005%, sulphuric acid
Ferrous 0.001%, sterilized water 97.794%;
The liquid aerobic bacteria culture medium prescription of described bacillus subtilis according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 1%, dipotassium hydrogen phosphate 0.05%, yeast extract 0.05%, Carnis Bovis seu Bubali cream 0.02%, peptone 0.05%, calcium carbonate
0.025%, sodium chloride 0.02%, manganese sulfate 0.001%, sterilized water 98.784%;
The liquid aerobic bacteria culture medium prescription of described Bacillus licheniformis according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 1%, dipotassium hydrogen phosphate 0.05%, yeast extract 0.05%, Carnis Bovis seu Bubali cream 0.02%, peptone 0.05%, calcium carbonate
0.025%, sodium chloride 0.02%, manganese sulfate 0.001%, sterilized water 98.784%;
The liquid anaerobic bacterium culture medium formula of described photosynthetic bacteria is: the 3.3g Han sodium acetate, ammonium chloride 0.6g in every 1 liter of sterilized water,
Magnesium sulfate 0.5g, yeast extract 1.5g;
The liquid anaerobic bacterium culture medium formula of described Lactobacillus plantarum is: the 20g Han sucrose in every 1 liter of sterilized water, yeast extract
25g, dipotassium hydrogen phosphate 2g;
The liquid anaerobic bacterium culture medium formula of described bacillus acidophilus is: the 5g Han sodium acetate in every 1 liter of sterilized water, ammonium citrate
2g, magnesium sulfate 0.5g, yeast extract 6g, Carnis Bovis seu Bubali cream 12g, dipotassium hydrogen phosphate 2g, peptone 10g, manganese sulfate 0.05g;
The liquid anaerobic bacterium culture medium formula of described Deshi Lactobacillus is: the 50g Han whey powder in every 1 liter of sterilized water, calcium carbonate
2g, yeast extract 10g;
Described mixed fermentation liquid culture based formulas is composed of the following components according to the weight/mass percentage composition shared by each component: sucrose
4%, yeast extract 0.4%, dipotassium hydrogen phosphate 0.2%, sterilized water 95.4%.
The microbial liquid microbial inoculum of embodiment 2 contrast experiment in yellow peach is planted:
Experimental period and place: in May, 2014 ~ October, lake, high and new technology industrial development zone town, Suzhou City spun gold yellow peach Specialty Co-operative Organization
Performance: randomly choose 10, the old peach tree of more than the 10 years age of trees as A community.Other 150 mu of peach trees are little as B
District.
Fertilising situation: A, B district all then 2, March use the every strain of base manure 10kg/ (total nutrient 3x15 compound fertilizer).May
Part every, A community peach tree microbial liquid microbial inoculum 100ml of embodiment 2, with pouring root and branch and leaf spray after 100 times of clear water dilutions
Execute.B uses community 5% dipotassium hydrogen phosphate solution and sprays and pouring root.
Experimental result: averagely every, A community result 410, jade-like stone profit that each Fructus Persicae is very large, commodity Fructus Persicae per cent pack reaches 96%.B
Averagely every, community result 322, the head of Fructus Persicae is less than normal than A community, and commodity Fructus Persicae per cent pack is less than 80%.
Above-described embodiment, only for technology design and the feature of the explanation present invention, its object is to allow person skilled in the art
Scholar will appreciate that present disclosure and implements according to this, can not limit the scope of the invention with this.All according to the present invention
The equivalence that spirit is made changes or modifies, and all should contain within protection scope of the present invention.
Claims (5)
1. the microbial liquid microbial inoculum that an aerobic bacteria is compound with anaerobe, it is characterised in that: described microbial liquid microbial inoculum bag
Include 6 kinds of aerobic microbial inoculums of liquid cultivated with fluid medium and 4 kinds of liquid anaerobic microbial inoculums cultivated with fluid medium, described liquid
The aerobic microbial inoculum of body is 3:7 ~ 7:3 with the mass ratio of liquid anaerobic microbial inoculum;The aerobic microbial inoculum of described liquid is by the microbial inoculum of following mass parts
Composition:
Liquid bacillus megaterium agent 1 ~ 3 mass parts,
Liquid blown-ball Azotobacter agent 1 ~ 3 mass parts,
Liquid colloid bacillus cereus agent 1 ~ 3 mass parts,
Liquid streptomyces microflavus agent 1 mass parts,
Liquid bacillus subtilis microbial inoculum 1 ~ 2 mass parts,
Liquid Bacillus licheniformis agent 1 ~ 2 mass parts;
Described liquid anaerobic microbial inoculum is made up of the microbial inoculum of following mass parts:
Liquid photosynthetic microbial inoculum 1 ~ 2 mass parts,
Liquid plant breast microbial inoculum 1 ~ 2 mass parts,
Liquid bacillus acidophilus's agent 1 ~ 5 mass parts,
Liquid Deshi Lactobacillus agent 1 ~ 5 mass parts;
Wherein, the good bacteria content in the aerobic microbial inoculum of each liquid is more than or equal to 200,000,000/milliliter, each liquid anaerobic bacterium
Anaerobe content in agent is more than or equal to 200,000,000/milliliter.
The microbial liquid microbial inoculum that a kind of aerobic bacteria the most according to claim 1 is compound with anaerobe, it is characterised in that: institute
State microbial liquid microbial inoculum and also include trace element, the addition of trace element be described microbial liquid microbial inoculum quality 0.1 ~
0.2%, described trace element is selected from B, Zn, Mn, Cu, Co the mixture of more than any one or at least two.
3. a preparation method for the microbial liquid microbial inoculum that aerobic bacteria according to claim 1 is compound with anaerobe, its
It is characterised by: described preparation method comprises the following steps:
6 kinds of aerobic bacterias are cultivated and are made 6 kinds of aerobic microbial inoculums of liquid by the first step respectively,
(1) prepare respectively bacillus megaterium, blown-ball Azotobacter, colloid bacillus cereus, streptomyces microflavus, bacillus subtilis with
And the liquid aerobic bacteria culture medium of Bacillus licheniformis, more each self-corresponding to 6 kinds of liquid aerobic bacteria inoculation of mediums respectively
Strain, at 30 ~ 37 DEG C, 120 ~ 140r/min, pH value carries out the one-level amplification culture of strain under the conditions of 6.5 ~ 7.5, obtains 6 kinds
The aerobic bacterium solution of one-level;
(2) 6 kinds of aerobic bacterium solution of one-level are taken the most respectively to 6 kinds of described each self-corresponding strains of liquid aerobic bacteria inoculation of medium,
30 ~ 37 DEG C, carry out two grades of amplification culture of strain under the conditions of 120 ~ 140r/min, obtain 6 kinds of two grades of aerobic bacterium solution, wherein,
The inoculum concentration of every kind of aerobic bacterium solution of one-level is the 0.9 ~ 1.1% of its liquid aerobic bacteria culture medium quality;
(3) 6 kinds of two grades of aerobic bacterium solution are taken again to 6 kinds of described each self-corresponding strains of liquid aerobic bacteria inoculation of medium, 30 ~
37 DEG C, carry out three grades of amplification culture of strain under the conditions of 120 ~ 140r/min, obtain 6 kinds of aerobic microbial inoculums of liquid, wherein, every kind two
Inoculum concentration is its liquid aerobic bacteria culture medium quality the 1 ~ 1.5% of the aerobic bacterium solution of level;
The liquid anaerobic microbial inoculum of 4 kinds of mixing is made in 4 kinds of anaerobic culturees by second step,
(1) the liquid anaerobic bacterium culture medium of photosynthetic bacteria, Lactobacillus plantarum, bacillus acidophilus and Deshi Lactobacillus is prepared respectively,
In 4 kinds of liquid anaerobic bacterium culture mediums, inoculate each self-corresponding strain the most respectively, stand under the conditions of 30 ~ 37 DEG C, carry out strain
One-level amplification culture, obtain 4 kinds of one-level anaerobism bacterium solution;
(2) take 4 kinds of one-level anaerobism bacterium solution the most respectively in 4 kinds of described liquid anaerobic bacterium culture mediums, inoculate each self-corresponding strain,
Stand under the conditions of 30 ~ 37 DEG C, carry out two grades of amplification culture of strain, obtain 4 kinds of secondary anaerobic bacterium solution, wherein, every kind of one-level
The inoculum concentration of anaerobism bacterium solution is the 1 ~ 1.5% of its liquid anaerobic bacterium culture medium quality;
(3) preparation mixed fermentation fluid medium, the formula proportion according to claim 1 takes described 4 kinds of secondary anaerobic bacterium solution to institute
State in mixed fermentation fluid medium inoculation mixing, 30 ~ 37 DEG C, stand mixed culture under the conditions of illumination 2800 ~ 3200Ix,
To the liquid anaerobic microbial inoculum of mixing, wherein, total inoculum concentration of 4 kinds of secondary anaerobic bacterium solution is described mixed fermentation liquid culture substrate
The 1 ~ 1.5% of amount;
3rd step, is slowly injected into the liquid anaerobic of described mixing by 6 kinds of aerobic microbial inoculums of liquid formula proportion according to claim 1
In microbial inoculum and stir, stand, behind isotropic stable, obtain described microbial liquid microbial inoculum.
The preparation method of the microbial liquid microbial inoculum that aerobic bacteria the most according to claim 3 is compound with anaerobe, its feature
Being: in described 3rd step, be simultaneously introduced trace element, the addition of trace element is described microbial liquid microbial inoculum quality
0.1 ~ 0.2%, described trace element more than any one or at least two mixture in B, Zn, Mn, Cu, Co.
The preparation method of the microbial liquid microbial inoculum that aerobic bacteria the most according to claim 3 is compound with anaerobe, its feature
Be: the liquid aerobic bacteria culture medium prescription of described bacillus megaterium according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 1%, dipotassium hydrogen phosphate 0.1%, ammonium sulfate 0.2%, soluble starch 2%, calcium carbonate 0.5%, Oleum Glycines 0.5%, nothing
Bacterium water 95.7%;
The liquid aerobic bacteria culture medium prescription of described blown-ball Azotobacter according to the weight/mass percentage composition shared by each component by following group
It is grouped into: sucrose 1%, dipotassium hydrogen phosphate 0.1%, yeast extract 0.5%, magnesium sulfate 0.05%, calcium carbonate 0.2%, sodium molybdate 0.001%,
Borax 0.001%, ferrous sulfate 0.001%, sodium chloride 0.002%, sterilized water 98.145%;
The liquid aerobic bacteria culture medium prescription of described colloid bacillus cereus according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 2%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.04%, yeast extract 0.2%, calcium carbonate 0.2%, ferrous sulfate
0.001%, sodium chloride 0.04%, sterilized water 97.419%;
The liquid aerobic bacteria culture medium prescription of described streptomyces microflavus according to the weight/mass percentage composition shared by each component by following group
It is grouped into: soluble starch 2%, dipotassium hydrogen phosphate 0.05%, potassium nitrate 0.1%, magnesium sulfate 0.05%, sodium chloride 0.005%, sulphuric acid
Ferrous 0.001%, sterilized water 97.794%;
The liquid aerobic bacteria culture medium prescription of described bacillus subtilis according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 1%, dipotassium hydrogen phosphate 0.05%, yeast extract 0.05%, Carnis Bovis seu Bubali cream 0.02%, peptone 0.05%, calcium carbonate
0.025%, sodium chloride 0.02%, manganese sulfate 0.001%, sterilized water 98.784%;
The liquid aerobic bacteria culture medium prescription of described Bacillus licheniformis according to the weight/mass percentage composition shared by each component by following
Component forms: sucrose 1%, dipotassium hydrogen phosphate 0.05%, yeast extract 0.05%, Carnis Bovis seu Bubali cream 0.02%, peptone 0.05%, calcium carbonate
0.025%, sodium chloride 0.02%, manganese sulfate 0.001%, sterilized water 98.784%;
The liquid anaerobic bacterium culture medium formula of described photosynthetic bacteria is: the 3.3g Han sodium acetate, ammonium chloride 0.6g in every 1 liter of sterilized water,
Magnesium sulfate 0.5g, yeast extract 1.5g;
The liquid anaerobic bacterium culture medium formula of described Lactobacillus plantarum is: the 20g Han sucrose in every 1 liter of sterilized water, yeast extract
25g, dipotassium hydrogen phosphate 2g;
The liquid anaerobic bacterium culture medium formula of described bacillus acidophilus is: the 5g Han sodium acetate in every 1 liter of sterilized water, ammonium citrate
2g, magnesium sulfate 0.5g, yeast extract 6g, Carnis Bovis seu Bubali cream 12g, dipotassium hydrogen phosphate 2g, peptone 10g, manganese sulfate 0.05g;
The liquid anaerobic bacterium culture medium formula of described Deshi Lactobacillus is: the 50g Han whey powder in every 1 liter of sterilized water, calcium carbonate
2g, yeast extract 10g;
Described mixed fermentation liquid culture based formulas is composed of the following components according to the weight/mass percentage composition shared by each component: sucrose
4%, yeast extract 0.4%, dipotassium hydrogen phosphate 0.2%, sterilized water 95.4%.
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