CN104557336A - Microbial fertilizer, preparation method and application thereof - Google Patents
Microbial fertilizer, preparation method and application thereof Download PDFInfo
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- CN104557336A CN104557336A CN201410844180.1A CN201410844180A CN104557336A CN 104557336 A CN104557336 A CN 104557336A CN 201410844180 A CN201410844180 A CN 201410844180A CN 104557336 A CN104557336 A CN 104557336A
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F3/00—Fertilisers from human or animal excrements, e.g. manure
- C05F3/02—Guano
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Abstract
The invention provides a microbial fertilizer, a preparation method and an application thereof. The microbial fertilizer comprises fungus species LP-20 and organic fertilizers, wherein the organic fertilizers comprise humic acid, turfy soil and composted chicken manure. The preparation method of the microbial fertilizer comprises the following steps: culturing the fungus species LP-20 to obtain a bacterial suspension; concentrating the obtain bacterial suspension to obtain a bacteria concentrate, and uniformly mixing the bacteria concentrate with the humic acid, the turfy soil and the composted chicken manure; pelletizing and drying to obtain the microbial fertilizer. The microbial fertilizer prepared from the fungus species LP-20 and the organic fertilizers can be applied to treatment of heavy metal pollution of soil, can effectively enrich free cadmium ions and zinc ions in the soil, has double effect of being taken as a production-increasing and fertilizer-maintaining fertilizer and a natural conditioner for treating the heavy metal pollution of the soil, and is an environmental friendly and ecological-coordination microbial fertilizer for treating the heavy metal pollution of the soil.
Description
Technical field
The invention belongs to microbial technology field, relate to a kind of microbial-bacterial fertilizer, its preparation method and application thereof.
Background technology
In decades recently, the development of the threat mankind that various environmental problem is more and more serious, the pollutents such as various industrial gaseous waste, waste water, chemical fertilizer, agricultural chemicals all enter in its final place body soil, therefore soil environment quality is caused day by day to decline, especially heavy metal pollution of soil, becomes the common focus paid close attention to of government and society instantly already.
Because heavy-metal contaminated soil structure is poor, nutrient deficiency, heavy metal exists with the form that toxicity is stronger, thus affects plant-growth, and heavy metal ion to soil enzyme activities produce suppress or activation, and then affect soil nutrient discharge and crop to the absorption of nutrient.Cadmium is relatively rare heavy metal element, is typical dispersed elements, is mainly present in the non-ferrous metal mineral reserve such as copper, lead, zinc with the form of sulfide.Cadmium can be brought in soil by various approach by mankind's activity, and it can not or excrete after not easily being transformed by biological decomposition, can only transmit step by step along food chain, concentrated amplification in vivo, toxic effect will be produced to organism when running up to high level, thus health risk.Relevant cadmium pollution and bioavailability problems thereof are caused the whole world to pay close attention to, and the sixties in 20th century is exactly cadmium pollution in the major cause of " itai-itai disease " that remarkable ability basin, river, Japanese fuji county occurs.Therefore, the research of Cadmium in Soil pollution source, form, biological effectiveness and corresponding governing measure is extremely necessary.
For administering heavy metal pollution of soil, usually to add various modifying agent to improve the physico-chemical property of soil, thus reduce the activity of Cadmium in Soil.Except N, P, K fertilizer of necessity, conventional modifying agent comprises humic acid, lime, Phosphate minerals, iron and manganese oxides, flyash, active sludge and compost etc.These modifying agents itself can reduce the activity of heavy metal in soil, play an important role in plant stability, but also there are some defects, as administered, area is little, production efficiency is low, and easily causes secondary pollution in preparation process.
Recently, carried out both at home and abroad a large amount of utilization can the biology of enriching heavy metal to remove the research of heavy metal pollution of soil.The advantages such as the method has efficiently, economy, ecological coordination, become the focus in environmental research.
CN 102553904 A discloses a kind of biological renovation method of heavy-metal contaminated soil, this invention utilizes earthworm to repair slight, moderate copper, cadmium, plumbous combined contamination soil, but this invention for the cultivation of earthworm and the culture cycle of plant longer, process is complicated.CN 103880486 A discloses microbial-bacterial fertilizer and preparation method thereof and the method for administering heavy-metal contaminated soil, and this invents the microbial-bacterial fertilizer utilizing wormcast to make and administers heavy metal pollution of soil.CN 1490280A discloses a kind of microorganism multi-antibacterial manure and its preparation method and application, utilize different efficacies bacterium, microorganism multi-antibacterial manure that fungi is made is a kind of microbial fertilizer with nutrition, biological control effect, but is not directed to the improvement of heavy metal contamination.
Therefore, need in this area to develop a kind of microbial-bacterial fertilizer effectively can administering heavy metal pollution of soil.
Summary of the invention
For the deficiencies in the prior art, the object of the present invention is to provide a kind of microbial-bacterial fertilizer, its preparation method and application thereof.
For reaching this goal of the invention, the present invention by the following technical solutions:
In first, the invention provides a kind of microbial-bacterial fertilizer, described microbial-bacterial fertilizer comprises fungi strain LP-20 and organic fertilizer, wherein said fungi strain LP-20 (Trichoderma spirale), be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), CGMCC), address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, 100101, preservation date is on August 16th, 2013, and deposit number is CGMCC No.8038.
In microbial-bacterial fertilizer of the present invention, described organic fertilizer comprises humic acid, turfy soil and Chicken dung.
In second, the invention provides a kind of preparation method of microbial-bacterial fertilizer of the present invention, comprise the following steps:
(1) fungi strain LP-20 is cultivated, obtain bacteria suspension;
(2) bacteria suspension that step (1) obtains is carried out concentrated obtain concentration thing, described concentration thing and humic acid, turfy soil and Chicken dung are mixed;
(3) namely microbial-bacterial fertilizer is obtained after granulation, oven dry.
In the preparation method of microbial-bacterial fertilizer of the present invention, the cultivation of the described fungi strain LP-20 of step (1) comprises the following steps:
A () culture dish is cultivated: be aseptically inoculated on solid medium by fungi strain LP-20, at 20-30 DEG C, preferably 25 DEG C, cultivates 4-6 days, preferably 5 days;
B () first order seed is cultivated: bacterial classification step (a) cultivated aseptically is inoculated in liquid nutrient medium, at 20-30 DEG C, preferably 25 DEG C, cultivates 3-7 days, preferably 5 days, obtains first order seed;
C () secondary seed is cultivated: the inoculum size by the volume ratio of liquid nutrient medium being 5-15%, preferred 8-12%, more preferably 10%, described first order seed is inoculated in fermentor tank, aeration, cultivates 5-7 days, preferably 5.5-6.5 days, preferably 6 days further, obtained secondary seed;
(d) mixing fermentation culture: the inoculum size by the volume ratio of liquid nutrient medium being 5-20%, preferred 5-12%, more preferably 10%, is inoculated into secondary seed in fermentor tank, carries out high density fermentation cultivation, obtain the bacteria suspension of this bacterial classification.
In the cultivation of fungi strain LP-20, the described solid medium of step (a) comprises following component: yeast extract 1-3g/L, preferably 1.5g/L, sodium-acetate 1.5-4g/L, preferably 2.5g/L, agar 13-18g/L, preferably 15g/L, paraxin 0.25mg/L.
In the cultivation of fungi strain LP-20, the described liquid nutrient medium of step (b) comprises following component: peptone 8-12g/L, preferably 10g/L, glucose 15-25g/L, preferably 20g/L, paraxin 0.25mg/L.
In culture dish is cultivated, culture temperature can be 20 DEG C, 21 DEG C, 22 DEG C, 23 DEG C, 24 DEG C, 25 DEG C, 26 DEG C, 27 DEG C, 28 DEG C, 29 DEG C or 30 DEG C, incubation time can be 4 days, 100 hours, 4.5 days, 120 hours, 5 days, 130 hours, 5.5 days, 140 hours or 6 days, in solid medium used, yeast extract can be 1g/L, 1.1g/L, 1.2g/L, 1.3g/L, 1.4g/L, 1.5g/L, 1.6g/L, 1.7g/L, 1.8g/L, 1.9g/L, 2.0g/L, 2.2g/L, 2.4g/L, 2.6g/L, 2.8g/L or 3.0g/L, sodium-acetate can be 1.5g/L, 1.6g/L, 1.7g/L, 1.8g/L, 2.0g/L, 2.2g/L, 2.3g/L, 2.4g/L, 2.5g/L, 2.7g/L, 2.8g/L, 3.0g/L, 3.2g/L, 3.4g/L, 3.6g/L, 3.8g/L, 3.9g/L or 4g/L, agar can be 13g/L, 14g/L, 15g/L, 16g/L, 17g/L or 18g/L.
In first order seed is cultivated, culture temperature can be 20 DEG C, 21 DEG C, 22 DEG C, 23 DEG C, 24 DEG C, 25 DEG C, 26 DEG C, 27 DEG C, 28 DEG C, 29 DEG C or 30 DEG C; Incubation time can be 3 days, 80 hours, 3.5 days, 90 hours, 4 days, 4.5 days, 110 hours, 5 days, 5.5 days, 140 hours, 6 days, 6.5 days or 7 days; In liquid nutrient medium used, peptone can be 8g/L, 9g/L, 10g/L, 11g/L or 12g/L, and glucose can be 15g/L, 16g/L, 17g/L, 18g/L, 19g/L, 20g/L, 21g/L, 22g/L, 23g/L, 24g/L or 25g/L.
In secondary seed is cultivated, inoculum size can be 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% or 15% by the volume ratio of liquid nutrient medium; Incubation time can be 5 days, 130 hours, 5.5 days, 140 hours, 6 days, 150 hours, 6.5 days, 160 hours or 7 days.
In mixing fermentation culture, inoculum size can be 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% by the volume ratio of liquid nutrient medium.
In the preparation method of microbial-bacterial fertilizer of the present invention, concentrate to have been come by centrifugal or press filtration described in step (2), object is that the moisture removed in bacteria suspension obtains concentration thing; The mass ratio of step (2) described mixture is 0.1%-5% concentration thing, 10% humic acid, 50% turfy soil and 35%-39.9% Chicken dung; Wherein said filter cake can be 0.1%, 0.3%, 0.5%, 0.7%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5% or 5%, and Chicken dung can be 35%, 35.4%, 35.8%, 36%, 36.2%, 36.5%, 36.8%, 37%, 37.4%, 37.8%, 38%, 38.3%, 38.5%, 38.7%, 38.9%, 39%, 39.2%, 39.4%, 39.6%, 39.8% or 39.9%.
3rd aspect, the invention provides described microbial-bacterial fertilizer and administering the application in heavy metal pollution of soil, microbial-bacterial fertilizer prepared by the present invention can be used for administering the heavy metal contamination such as Cadmium in Soil and zinc.
Relative to prior art, the present invention has following beneficial effect:
Fungi strain LP-20 screening and separating of the present invention, from soil, belongs to Trichoderma, has biological safety, and the microbial-bacterial fertilizer containing this fungi strain can the effectively cadmium ion of free state and zine ion in Enriching soil, thus reduces the activity of Cadmium in Soil and zinc.Fertilizer in bacterial manure also has heavy metal adsorption characteristic, and this organic fertilizer can promote that bacterial classification LP-20 is to the adsorption of heavy metal in soil, and nutritive substance needed for plant-growth can be provided, fungi strain LP-20 and organic fertilizer are coordinated and forms microbial-bacterial fertilizer, more effectively can adsorb the cadmium in soil and zinc heavy metal.Therefore, microbial-bacterial fertilizer provided by the invention has the fertilizer serving as volume increase fertilizer conservation and the double effects of natural amendments administering heavy metal pollution of soil, is a kind of heavy metal pollution of soil improvement microbial-bacterial fertilizer of environmentally friendly, ecological coordination.
Accompanying drawing explanation
Fig. 1 adsorbs the design sketch of heavy metal cadmium and zinc in soil to LP-20 at the differential responses time;
Fig. 2 is that different soils pH value adsorbs the design sketch of heavy metal cadmium and zinc in soil to LP-20;
Fig. 3 be there is different bacterial classifications and organic fertilizer ratio of mixture microbial-bacterial fertilizer to the design sketch adsorbing heavy metal cadmium in soil.
Embodiment
Technical scheme of the present invention is further illustrated below by embodiment.Those skilled in the art should understand, described embodiment is only help to understand the present invention, should not be considered as concrete restriction of the present invention.
in embodiment 1 bacterial manure, fungi strain is to the adsorption effect of heavy metal in soil cadmium and zinc
In the present embodiment, by the following method fungi strain LP-20 is cultivated, comprises the following steps:
A () culture dish is cultivated: be aseptically inoculated on solid medium by fungi strain LP-20, cultivates 5 days at 25 DEG C;
B () first order seed is cultivated: bacterial classification step (a) cultivated aseptically is inoculated in liquid nutrient medium, cultivates 5 days at 25 DEG C, obtained first order seed;
C () secondary seed is cultivated: be the inoculum size of 10% by the volume ratio of liquid nutrient medium, is inoculated into by described first order seed in 5L fermentor tank, aeration, cultivates 6 days, obtains secondary seed;
D () mixing fermentation culture: be the inoculum size of 10% by the volume ratio of liquid nutrient medium, is inoculated into secondary seed in fermentor tank (mechanical stirring), carries out high density fermentation cultivation, obtain the bacteria suspension of this bacterial classification.
Different adsorption time adsorbs the impact of heavy metal cadmium and zinc in soil on fungi LP-20
Prepare 6 parts of equivalent 5g pedotheques (in sample, Heavy Metals is cadmium and zinc), every two parts of pedotheques are one group, often in group, portion is that blank is without any process, another part adds above-mentioned bacteria suspension 30mL fungi strain LP-20 being cultivated to rear acquisition, pH regulator is 7.0 (utilizing 1M HCl or 1MNaOH to regulate), incubated at room temperature.With repairing efficiency 10d (my god), 20d, 30d be that node gets Cd, Zn bio-available Zn concentration in one group of sample determination soil respectively, measurement result is as shown in Figure 1.
As can be seen from Figure 1, the fixed efficiency of LP-20 to cadmium (Cd (II)) just reaches about 95% after 10d, substantially remains on more than 95% afterwards at the fixed efficiency of 20d, 30d, does not occur significantly to increase.Therefore, LP-20 needs 10d for Cd (II) fixation response time.LP-20 but has certain change for the fixed effect of zinc (Zn (II)), the fixed effect of 10d only has about 70%, but when reacted between extend to 20d, fixed efficiency is increased to about 85%, a small amount of decline is had during 30d, but still more than 80%.Therefore, for utilizing LP-20 fixing the Zn (II) in soil, the reaction times is the most suitable at 20d.
Different soils pH value adsorbs the impact of heavy metal cadmium and zinc in soil to fungi LP-20
PH value is one of most important soil parameters, different pH value to the growth conditions of heavy metal occurrence patterns, migrate attribute and the fungal bacterial strain in soil all tool have a certain impact.Therefore, investigate the impact of different pH value on LP-20 fixation of Cd (II) and Zn (II) effect, to practical application from now on, there is important reference value.
Prepare 10 parts of equivalent 5g pedotheques, every two parts is one group, 5 groups of sample pH are adjusted to respectively 3,4,5,6,7 (utilizing 1mol/L HCl or 1mol/L NaOH), often in group, portion is that blank is without any process, another part adds above-mentioned bacteria suspension 30mL fungi strain LP-20 being cultivated to rear acquisition, incubated at room temperature, incubation time is 10d.Cultivate Cd, Zn bio-available Zn concentration terminated in rear mensuration pedotheque, result as shown in Figure 2.
As can be seen from Figure 2, under different pH value, the effect that LP-20 fixes two heavy metal species ions is different.LP-20 reaches maximum value 96.45% for the fixed efficiency of Cd (II) at pH=4, all remains on more than 95%, be down to less than 80% when dropping to less than 90%, pH=7 when pH=6 in pH3-5 scope internal fixtion efficiency.Therefore, the optimization pH scope utilizing the Cd (II) in LP-20 fixing soil is 3-5.LP-20 reaches maximum value 68.34% for the fixed efficiency of Zn (II) at pH=3, all remains on more than 65%, be down to less than 40% when dropping to about 40%, pH=7 when pH=6 in pH3-5 scope internal fixtion efficiency.Therefore, the optimization pH scope of the Zn in LP-20 fixing soil (II) is utilized to be similarly 3-5.In sum, the fixing optimal ph scope of LP-20 to Cd in soil (II), Zn (II) is 3-5.
the different blended composition and division in a proportion of embodiment 2 bacterial classification and organic fertilizer adsorbs heavy metal cadmium in soil to microbial-bacterial fertilizer
with the impact of zinc
First, prepare microbial-bacterial fertilizer sample 1#, 2#, 3#, 4#, 5# and 6#, wherein 1# sample preparation is as follows: cultivate as embodiment 1 and obtain LP-20 bacteria suspension, centrifugal treating is carried out to it, obtain concentrated thalline, get concentrated thalline 0.01g, itself and 1g humic acid, 5g turfy soil and 3.99g Chicken dung are mixed and makes microbial-bacterial fertilizer 1# sample (in microbial-bacterial fertilizer, thalline content is 0.1%); 2# sample preparation is as follows: cultivate as embodiment 1 and obtain LP-20 bacteria suspension, centrifugal treating is carried out to it, obtain concentrated thalline, get concentrated thalline 0.1g, itself and 1g humic acid, 5g turfy soil and 3.9g Chicken dung are mixed and makes microbial-bacterial fertilizer 2# sample (in microbial-bacterial fertilizer, thalline content is 1%); 3# sample preparation is as follows: cultivate as embodiment 1 and obtain LP-20 bacteria suspension, centrifugal treating is carried out to it, obtain concentrated thalline, get concentrated thalline 0.2g, itself and 1g humic acid, 5g turfy soil and 3.8g Chicken dung are mixed and makes microbial-bacterial fertilizer 3# sample (in microbial-bacterial fertilizer, thalline content is 2%); 4# sample preparation is as follows: cultivate as embodiment 1 and obtain LP-20 bacteria suspension, centrifugal treating is carried out to it, obtain concentrated thalline, get concentrated thalline 0.3g, itself and 1g humic acid, 5g turfy soil and 3.7g Chicken dung are mixed and makes microbial-bacterial fertilizer 4# sample (in microbial-bacterial fertilizer, thalline content is 3%); 5# sample preparation is as follows: cultivate as embodiment 1 and obtain LP-20 bacteria suspension, centrifugal treating is carried out to it, obtain concentrated thalline, get concentrated thalline 0.4g, itself and 1g humic acid, 5g turfy soil and 3.6g Chicken dung are mixed and makes microbial-bacterial fertilizer 5# sample (in microbial-bacterial fertilizer, thalline content is 4%); 6# sample preparation is as follows: cultivate as embodiment 1 and obtain LP-20 bacteria suspension, centrifugal treating is carried out to it, obtain concentrated thalline, get concentrated thalline 0.5g, itself and 1g humic acid, 5g turfy soil and 3.5g Chicken dung are mixed and makes microbial-bacterial fertilizer 6# sample (in microbial-bacterial fertilizer, thalline content is 5%).
Secondly, prepare control sample 0#, preparation method is as follows: control sample 0# is made in the mixing of 1g humic acid, 5g turfy soil and 4.0g Chicken dung.
Prepare 7 parts of equivalent 10g pedotheques, pH regulator is 7.0 (utilizing 1M HCl or 1M NaOH to regulate), and a copy of it is blank, other six parts of sample 5g adding numbering 0#, 1#, 2#, 3#, 4#, 5# and 6# respectively, incubated at room temperature, incubation time is 20d.Cultivate Cd available state (Cd (the II)) content terminated in rear mensuration pedotheque, measurement result as shown in Figure 3.
Can significantly see from Fig. 3, bacterial classification is different from the fixed efficiency of different blended composition and division in a proportion to Cadmium in Soil of organic fertilizer.Under the condition of adding separately organic fertilizer (0# sample), also have certain fixed efficiency to cadmium, fixed efficiency is about 65%, and this illustrates that the fertilizer in bacterial manure also has certain heavy metal adsorption characteristic.After bacterial classification mixes with organic fertilizer, the fixed efficiency of heavy metal cadmium strengthens, when thalline content is 2%, fixed efficiency can increase to about 88% (2# sample), analyze its reason, may be that the growth that organic fertilizer is microbial inoculum provides nutritional condition, microorganism in bacterial manure is bred, thus more effectively the heavy metal in soil is fixed, but along with bacterial classification addition continue increase, the fixed efficiency of cadmium is declined to some extent, this may be because cell concentration in bacterial manure increases, competition enhancing between the growth and breeding of thalline makes to decline to the fixed efficiency of cadmium, the best bacterial classification addition shown in this experiment is 1% and 2%.
Therefore, the present invention utilizes fungi strain LP-20 to coordinate with humic acid, turfy soil and Chicken dung and makes microbial-bacterial fertilizer, this bacterial manure can the effectively cadmium ion of free state and zine ion in Enriching soil, thus reduce the activity of Cadmium in Soil and zinc, in addition, fertilizer in bacterial manure also has heavy metal adsorption characteristic, and this organic fertilizer can promote that bacterial classification LP-20 is to the adsorption of heavy metal in soil, and can provide the nutritive substance needed for plant-growth.Therefore, microbial-bacterial fertilizer provided by the invention has the fertilizer serving as volume increase fertilizer conservation and the double effects of natural amendments administering heavy metal pollution of soil, is a kind of heavy metal pollution of soil improvement microbial-bacterial fertilizer of environmentally friendly, ecological coordination.
Applicant states, the present invention illustrates the present invention by above-described embodiment, but the present invention is not limited to above-described embodiment could realize.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of ancillary component, the concrete way choice etc. of raw material selected by the present invention, all drops within protection scope of the present invention and open scope.
Claims (9)
1. a microbial-bacterial fertilizer, it is characterized in that, described microbial-bacterial fertilizer comprises fungi strain LP-20 and organic fertilizer, wherein said fungi strain LP-20 (Trichoderma spirale), be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preservation date is on August 16th, 2013, and deposit number is CGMCC No.8038.
2. microbial-bacterial fertilizer according to claim 1, is characterized in that, described organic fertilizer comprises humic acid, turfy soil and Chicken dung.
3. the preparation method of microbial-bacterial fertilizer according to claim 1 and 2, is characterized in that, comprises the following steps:
(1) fungi strain LP-20 is cultivated, obtain bacteria suspension;
(2) bacteria suspension that step (1) obtains is carried out concentrated obtain concentration thing, described concentration thing and humic acid, turfy soil and Chicken dung are mixed;
(3) namely microbial-bacterial fertilizer is obtained after granulation, oven dry.
4. preparation method according to claim 3, is characterized in that, the cultivation of the described fungi strain LP-20 of step (1) comprises the following steps:
A () culture dish is cultivated: be aseptically inoculated on solid medium by fungi strain LP-20, at 20-30 DEG C, preferably 25 DEG C, cultivates 4-6 days, preferably 5 days;
B () first order seed is cultivated: bacterial classification step (a) cultivated aseptically is inoculated in liquid nutrient medium, at 20-30 DEG C, preferably 25 DEG C, cultivates 3-7 days, preferably 5 days, obtains first order seed;
C () secondary seed is cultivated: the inoculum size by the volume ratio of liquid nutrient medium being 5-15%, preferred 8-12%, more preferably 10%, described first order seed is inoculated in fermentor tank, aeration, cultivates 5-7 days, preferably 5.5-6.5 days, preferably 6 days further, obtained secondary seed;
(d) mixing fermentation culture: the inoculum size by the volume ratio of liquid nutrient medium being 5-20%, preferred 5-12%, more preferably 10%, is inoculated into secondary seed in fermentor tank, carries out high density fermentation cultivation, obtain the bacteria suspension of this bacterial classification.
5. preparation method according to claim 4, it is characterized in that, the described solid medium of step (a) comprises following component: yeast extract 1-3g/L, preferably 1.5g/L, sodium-acetate 1.5-4g/L, preferably 2.5g/L, agar 13-18g/L, preferably 15g/L, paraxin 0.25mg/L.
6. the preparation method according to claim 4 or 5, is characterized in that, the described liquid nutrient medium of step (b) comprises following component: peptone 8-12g/L, preferably 10g/L, glucose 15-25g/L, preferably 20g/L, paraxin 0.25mg/L.
7. the preparation method according to any one of claim 4-6, is characterized in that, in step (2) mixture, the mass ratio of each component is 0.1%-5% concentration thing, 10% humic acid, 50% turfy soil and 35%-39.9% Chicken dung.
8. the microbial-bacterial fertilizer that microbial-bacterial fertilizer according to claim 1 and 2 or the preparation method according to any one of claim 3-7 prepare is administering the application in heavy metal pollution of soil.
9. application according to claim 8, is characterized in that, described microbial-bacterial fertilizer is for administering Cadmium in Soil and zinc heavy metal contamination.
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| CN107058123A (en) * | 2017-03-10 | 2017-08-18 | 中国海洋大学 | A kind of Aureobasidium pullulans and its application |
| CN107641049A (en) * | 2017-09-22 | 2018-01-30 | 济南润丰伟业生物科技有限公司 | A kind of microbiological fermented organic fertilizer of repairing heavy metal in soil pollution |
| CN114075438A (en) * | 2021-12-02 | 2022-02-22 | 上海康恒环境修复有限公司 | Composite biological agent for repairing heavy metal contaminated soil and preparation method and application thereof |
| CN118020796A (en) * | 2024-03-04 | 2024-05-14 | 江苏农林职业技术学院 | Application of trichoderma spirochete JSAFC 2090,2090 |
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