CN108130304A - Bacillus composite bacterial liquid and preparation method thereof - Google Patents
Bacillus composite bacterial liquid and preparation method thereof Download PDFInfo
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- CN108130304A CN108130304A CN201810137292.1A CN201810137292A CN108130304A CN 108130304 A CN108130304 A CN 108130304A CN 201810137292 A CN201810137292 A CN 201810137292A CN 108130304 A CN108130304 A CN 108130304A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- 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
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- 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
- C09K2101/00—Agricultural use
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- 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
- C09K2109/00—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE pH regulation
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Abstract
The invention provides a bacillus composite bacterial liquid, which comprises the following components: bacillus subtilis, bacillus licheniformis and bifidobacterium. Wherein the preservation number of the bacillus subtilis is CGMCC1.3358, the preservation number of the bacillus licheniformis is ATCC 14580, and the preservation number of the bifidobacterium is ATCC 15707; the viable count of the bacillus subtilis, the bacillus licheniformis and the bifidobacterium is respectively 108～109CFU/g. The invention also provides a preparation method of the bacillus composite bacterial liquid, the operation steps of the method are simple, and the composite bacterial liquid can promote microorganisms to decompose organic matters to form humus, so that the effect of improving soil is achieved, and the absorption and growth of plants on nutrient substances are promoted. Has the effects of protecting soil, promoting the decomposition of organic matters, generating rich metabolites, inhibiting harmful bacteria and the like, thereby playing the roles of growing and protecting plants.
Description
Technical field
The invention belongs to field of biological, and in particular to a kind of bacillus composite bacteria liquid and preparation method thereof.
Background technology
Bacillus subtilis and bacillus licheniformis, be bacterium a section, can be formed gemma (endospore) bacillus or
Coccus is strong to extraneous injurious factor resistance, and distribution is wide, is present in soil, water, air and animal intestinal tract etc..Adjustable bacterium
Group's imbalance reaches therapeutic purposes, and body can be promoted to generate antibacterial substance, kill pathogenic bacteria.Resistant activity material can be generated, and
With unique biology take oxygen mechanism, the growth and breeding of pathogenic bacteria can be inhibited.The bacterium of Bifidobacterium is humans and animals
One of important composition member of intestinal flora is a kind of important intestinal beneficial microbes, can inhibit harmful levels of pathogens, play biology
Preventive and therapeutic effect.
Humus in the soil is that organic matter converts the colloidal substance to be formed, generally black or dark through microorganism decomposition
Brown is the chief component of the soil organism.Currently, due to it is unreasonable using chemical fertilizer, it is excessive cultivation, single
Soil degradation problem caused by cropping system serious threat to agricultural sustainable development, wherein, the humus in soil
Reduction is major reason.
Invention content
In view of this, the purpose of the present invention is to provide a kind of bacillus composite bacteria liquid, including following components：Withered grass bud
Spore bacillus, bacillus licheniformis and Bifidobacterium.
Preferably, in bacillus composite bacteria liquid of the present invention, the bacillus subtilis is commonly used in the art
Bacillus subtilis, it is preferable that the deposit number of the bacillus subtilis be CGMCC1.3358.
Preferably, in bacillus composite bacteria liquid of the present invention, the bacillus licheniformis is commonly used in the art
Bacillus licheniformis, it is preferable that the deposit number of the bacillus licheniformis be ATCC14580.
Preferably, in bacillus composite bacteria liquid of the present invention, the Bifidobacterium is pair commonly used in the art
Discrimination bacillus, it is preferable that the deposit number of the Bifidobacterium is ATCC 15707.
Preferably, in bacillus composite bacteria liquid of the present invention, the bacillus subtilis, bacillus licheniformis with
The viable count of Bifidobacterium is respectively 108~109CFU/g。
Another aspect of the invention is a kind of preparation method of bacillus composite bacteria liquid is provided, include the following steps：
1) Bacillus subtilis strain accesses fluid nutrient medium after activation, add in bifidobacteria viable bacteria powder, sealing isolation sky
Air culture is supported；
2) bacillus licheniformis will be activated, accesses fluid nutrient medium culture；
3) merge step 1) and two step bacterium solution of step 2), mixed bacteria liquid concentration centrifugation；
4) mixing is concentrated into centrifugation bacterium solution and adds in nutrient solution to get bacillus composite bacteria liquid.
Preferably, in the preparation method of bacillus composite bacteria liquid of the present invention, the guarantor of the bacillus subtilis
It is CGMCC 1.3358 to hide number；The deposit number of the bacillus licheniformis is ATCC 14580.
Preferably, in the preparation method of bacillus composite bacteria liquid of the present invention, the bacillus subtilis, lichens
The viable count of bacillus and Bifidobacterium is respectively 108~109CFU/g。。
Preferably, in the preparation method of bacillus composite bacteria liquid of the present invention, condition of culture in the step 1)
It it is 35~39 DEG C, 120~130rpm cultivates 15~20h.
Preferably, in the preparation method of bacillus composite bacteria liquid of the present invention, condition of culture in the step 2)
It it is 35~39 DEG C, 120~130rpm cultivates 15~20h.
Compared with prior art, the present invention the present invention has the following advantages：
The growth of Bifidobacterium is stringenter to environmental requirement, and in particular for strictly anaerobic, oxygen is removed by needing
It is cultivated, can not be directly used in soil conditioner；The present invention is first cultivated by the optimization to culture medium and condition of culture
Bacillus subtilis so that bacillus subtilis forms anaerobic condition in growth consumption, promotes bifidobacterium growth so that gemma
Bacillus forms symbiosis with Bifidobacterium, then is cultivated by bacillus licheniformis, further has adjusted ecological flora so that compound bacteria
Liquid can promote microorganism decomposition organic matter, form humus, by improving the physicochemical property of soil, break soil hardening and pressure
It is real, promote the decomposition of the soil organism and the release of nutritional ingredient, improve soil control fertilizer ability, adjust soil pH, adjust micro- life
Object Bacterial community inhibits harmful microbe growth, enhances root system of plant to the absorbability of nutriment and promote growth,
Reach Effect of improving soil, and then with the abundant metabolin of soil protection, the decomposition of promotion organic matter, generation and inhibit harmful
The effects that bacterium, so as to play the growth and protective effect to plant, promotes the absorption and growth of Plant To Nutrient substance.By rear
Continuous embodiment proves that the composite bacteria liquid of the present invention has good volume increase effect of gain.
Specific embodiment
In an embodiment of the invention, the deposit number of bacillus subtilis of the present invention is CGMCC
1.3358。
In an embodiment of the invention, the deposit number of bacillus licheniformis of the present invention is ATCC
14580。
In an embodiment of the invention, the deposit number of Bifidobacterium of the present invention is ATCC 15707.
In an embodiment of the invention, bacillus subtilis of the present invention, bacillus licheniformis and bifid bar
The viable count of bacterium is respectively 108~109CFU/g。
In an embodiment of the invention, the present invention provides for example a kind of preparation side of bacillus composite bacteria liquid
Method includes the following steps：
1) by Bacillus subtilis strain, fluid nutrient medium is accessed after activation, adds in bifidobacteria viable bacteria powder, sealing isolation
Air jet flow；
2) bacillus licheniformis will be activated, accesses fluid nutrient medium culture；
3) merge step 1) and two step bacterium solution of step 2), mixed bacteria liquid concentration centrifugation；
4) mixing is concentrated into centrifugation bacterium solution and adds in nutrient solution to get bacillus composite bacteria liquid.
In an embodiment of the invention, the nutrient solution is preferably plant growth nutrient fluid, more preferably
Commercially available Aron-Hoagland nutrient solutions；The volume basis of nutrient solution contains in heretofore described bacillus composite bacteria liquid
Amount preferably 45~55%, more preferably 50%.
In an embodiment of the invention, in the preparation method of bacillus composite bacteria liquid of the present invention, institute
The deposit number for stating bacillus subtilis is CGMCC 1.3358；The deposit number of the bacillus licheniformis is ATCC
14580, the deposit number of the Bifidobacterium is ATCC 15707.
In an embodiment of the invention, in the preparation method of bacillus composite bacteria liquid of the present invention, institute
The viable count for stating bacillus subtilis, bacillus licheniformis and Bifidobacterium in composite bacteria liquid product is respectively 108~109CFU/
g。
In an embodiment of the invention, in the preparation method of bacillus composite bacteria liquid of the present invention, institute
It is 35~39 DEG C to state condition of culture in step 1), and 120~130rpm cultivates 15~20h.
In an embodiment of the invention, in the preparation method of bacillus composite bacteria liquid of the present invention, institute
It is 35~39 DEG C to state condition of culture in step 2), and 120~130rpm cultivates 15~20h.
Below in conjunction with the embodiment in the present invention, the technical solution in the present invention is clearly and completely described.It is aobvious
So, described embodiment is only part of the embodiment of the present invention, instead of all the embodiments.Based on the reality in the present invention
Apply example, those of ordinary skill in the art's all other embodiments obtained without making creative work all belong to
In the scope of protection of the invention.
Embodiment 1
1) bacterial strain explanation：The deposit number of bacillus subtilis for CGMCC 1.3358 and bacillus licheniformis preservation
Number is ATCC 14580, and the deposit number of Bifidobacterium is ATCC 15707；More than bacterial strain is to be obtained commercially.
2) bacillus is cultivated with Bifidobacterium：The Bacillus subtilis strain that 2 rings is taken to activate, access loading amount are 100mL
In the 500mL triangular flasks of fluid nutrient medium, and 0.1g bifidobacteria viable bacterias powder (being obtained commercially) is added in, triangular flask is sealed, into
Row Anaerobic culturel.By optimizing fluid nutrient medium and condition of culture, wherein basal liquid medium composition for beef extract 3.0g (i.e.
0.3%), peptone 10.0g (1%), sodium chloride 5.0g (0.5%), distilled water 1000ml.By in optimization culture culture medium
Beef extract 0.2~0.8%, most preferably 0.4%；Peptone selects yeast extract powder to replace and is used as nitrogen source 0.5~1%, most
It is preferably in a proportion of 0.9%；Sodium chloride 0.1~0.5% is most preferably 0.3%.Consumption is grown using bacillus subtilis to be formed
Anaerobic condition using bifidobacterium growth, forms symbiosis, and bacillus subtilis and Bifidobacterium are mixed its best culture item
Part pH 7.0~7.5, optimal pH are 7.2；Rotating speed is cultivated in 130~160rpm, optimal is 150rpm；Optimum temperature 35~38
DEG C, Optimal Temperature condition is 37 DEG C.By culture 18~for 24 hours, bacillus reaches 108~1010CFU/g, bifidobacterium concentration
108~1010CFU/g。
3) bacillus licheniformis is cultivated：The Bacillus licheniformis strain that 2 rings is taken to activate, access loading amount are trained for 100mL liquid
In the 500mL triangular flasks for supporting base, by optimizing fluid nutrient medium and condition of culture, wherein basal liquid medium composition is beef
Cream 3.0g (i.e. 0.3%), peptone 10.0g (1%), sodium chloride 5.0g (0.5%), distilled water 1000ml.Pass through optimization culture
Beef extract 0.2~0.8% in culture medium, most preferably 0.4%；Peptone selects yeast extract powder to replace as nitrogen source 0.5
~1%, most preferred ratio 0.9%；Sodium chloride 0.1~0.5% is most preferably 0.3%.It is given birth to using bacillus subtilis
Long consumption forms anaerobic condition, using bifidobacterium growth, forms symbiosis, optimal culture condition pH7.0~7.5, optimal pH
It is 7.2；Rotating speed is cultivated in 130~160rpm, optimal is 150rpm；35~38 DEG C of optimum temperature, Optimal Temperature condition are 37 DEG C.
By culture 18~for 24 hours, concentration reaches 108~1010CFU/g。
4) by above two steps bacterium solution according to 1：1 ratio merges, and obtains 200ml mixed bacteria liquids, and 150ml is centrifuged by concentration
Bacterium solution.
5) 150ml is mixed into concentration centrifugation bacterium solution and presses 1:It is compound to get 300ml bacillus to add in nutrient solution for 1 volume ratio
The viable count of bacterium solution, wherein bacillus subtilis, bacillus licheniformis and Bifidobacterium is respectively 108~109CFU/g。
The composite bacteria liquid of bacillus subtilis, bacillus licheniformis and Bifidobacterium that embodiment 1 obtains, with 3 kinds of withered grass
Bacillus, bacillus licheniformis increase agron compared with the independent bacterium solution (identical viable count) of Bifidobacterium
20%, so as to achieve the effect that improve soil, found by cultivating same breed potato, the bacillus subtilis that the present invention obtains
The composite bacteria liquid of bacterium, bacillus licheniformis and Bifidobacterium, with 3 kinds of bacillus subtilises, bacillus licheniformis and Bifidobacterium
Independent bacterium solution compare, potato weightening 40%.It is found by cultivating same breed wheat, the bacillus subtilis that the present invention obtains
The composite bacteria liquid of bacterium, bacillus licheniformis and Bifidobacterium, with 3 kinds of bacillus subtilises, bacillus licheniformis and Bifidobacterium
Arbitrary two kinds of bacterium solutions compare, wheat weightening more than 20%.It is found by cultivating same breed tomato, the present invention obtains withered
The composite bacteria liquid of careless bacillus, bacillus licheniformis and Bifidobacterium, compared with commercially available farm manure, weightening 16%.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of bacillus composite bacteria liquid, including following components：Bacillus subtilis, bacillus licheniformis and bifid bar
Bacterium.
2. bacillus composite bacteria liquid according to claim 1, which is characterized in that the preservation of the bacillus subtilis is compiled
Number be CGMCC 1.3358.
3. bacillus composite bacteria liquid according to claim 1, which is characterized in that the preservation of the bacillus licheniformis is compiled
Number be ATCC 14580.
4. bacillus composite bacteria liquid according to claim 1, which is characterized in that the deposit number of the Bifidobacterium is
ATCC 15707。
5. bacillus composite bacteria liquid according to claims 2 to 4, which is characterized in that the bacillus subtilis, lichens
The viable count of bacillus and Bifidobacterium is respectively 108~109CFU/g。
6. a kind of preparation method of bacillus composite bacteria liquid, includes the following steps：
1) by Bacillus subtilis strain, fluid nutrient medium is accessed after activation, adds in bifidobacteria viable bacteria powder, sealing isolation air
Culture；
2) by bacillus licheniformis, fluid nutrient medium culture is accessed；
3) merge step 1) and two step bacterium solution of step 2), mixed bacteria liquid concentration centrifugation；
4) mixing is concentrated into centrifugation bacterium solution and adds in nutrient solution to get bacillus composite bacteria liquid.
7. the preparation method of bacillus composite bacteria liquid according to claim 6, which is characterized in that the bacillus subtilis
The deposit number of bacterium is CGMCC 1.3358；The deposit number of the bacillus licheniformis is ATCC 14580；The bifid bar
The deposit number of bacterium is ATCC 15707.
8. the preparation method of bacillus composite bacteria liquid according to claim 7, which is characterized in that the bacillus subtilis
The viable count of bacterium, bacillus licheniformis and Bifidobacterium is respectively 108~109CFU/g。。
9. the preparation method of bacillus composite bacteria liquid according to claim 7, which is characterized in that trained in the step 1)
The condition of supporting is 35~39 DEG C, and 120~130rpm cultivates 15~20h.
10. the preparation method of bacillus composite bacteria liquid according to claim 7, which is characterized in that in the step 2)
Condition of culture is 35~39 DEG C, and 120~130rpm cultivates 15~20h.
Priority Applications (1)
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| CN201810137292.1A CN108130304A (en) | 2018-02-10 | 2018-02-10 | Bacillus composite bacterial liquid and preparation method thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109518703A (en) * | 2019-01-15 | 2019-03-26 | 内蒙古工业大学 | A kind of administering method of the cementing treatment fluid of microbial mineralization and chiltern side slope |
| CN109540615A (en) * | 2018-11-09 | 2019-03-29 | 中国科学院东北地理与农业生态研究所 | A kind of mud cultivation measuring soil organic carbon mineralization |
| CN110818508A (en) * | 2019-12-04 | 2020-02-21 | 河南农贝得农业科技有限公司 | Microbial agent with function of breaking soil hardening and preparation method thereof |
| CN114616942A (en) * | 2022-04-09 | 2022-06-14 | 中南林业科技大学 | Carbon sink system plants and uses soil treatment device |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109540615A (en) * | 2018-11-09 | 2019-03-29 | 中国科学院东北地理与农业生态研究所 | A kind of mud cultivation measuring soil organic carbon mineralization |
| CN109518703A (en) * | 2019-01-15 | 2019-03-26 | 内蒙古工业大学 | A kind of administering method of the cementing treatment fluid of microbial mineralization and chiltern side slope |
| CN109518703B (en) * | 2019-01-15 | 2021-07-02 | 内蒙古工业大学 | Microbial mineralization and cementation treatment fluid and treatment method of sandy slope |
| CN110818508A (en) * | 2019-12-04 | 2020-02-21 | 河南农贝得农业科技有限公司 | Microbial agent with function of breaking soil hardening and preparation method thereof |
| CN114616942A (en) * | 2022-04-09 | 2022-06-14 | 中南林业科技大学 | Carbon sink system plants and uses soil treatment device |
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