CN106754449A - A kind of utilization cow dung or horsehit prepare technology and its application of liquid bio-bacterial manure - Google Patents
A kind of utilization cow dung or horsehit prepare technology and its application of liquid bio-bacterial manure Download PDFInfo
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- CN106754449A CN106754449A CN201510798467.XA CN201510798467A CN106754449A CN 106754449 A CN106754449 A CN 106754449A CN 201510798467 A CN201510798467 A CN 201510798467A CN 106754449 A CN106754449 A CN 106754449A
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- horsehit
- cow dung
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
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Abstract
The invention belongs to biological technical field, it is culture matrix using cow dung or horsehit to disclose a kind of, using multiple-microorganism strain, according to the technology of certain formula rate fermenting and preparing biological Liquid Fertilizer in bioreactor.Biological liquid fertilizer can increase soil fertility, and improve utilization and conversion of the long-term cropping to required nutrient, increase plantation yield.Another purposes of biological liquid fertilizer can also significantly inhibit pathogenic microorganism growth, reduce long-term cropping disease and occur, and reduce Pesticide use.Pseudomonas stutzeri and bacillus amyloliquefaciens are by multiple strain improvement in the prominent present invention, and can all carry out liquid deep layer fermenting large-scale production.
Description
Technical field
The invention belongs to biological technical field, it is related to two plants of soil beneficial bacteriums and its enzyme, bacterial strain is Pseudomonas stutzeri
（Pseudomonas stutzeri JF-1）With bacillus amyloliquefaciens (Bacillus amyloliquefaciens JF-2).
Background technology
Narrow senseBio-feritlizer, it is to pass throughMicroorganismVital movement, makes crops obtain the product of specific fertilizer effect,
Also be referred to as Inoculant orBacterial manure, itself is without nutrient, it is impossible to instead of chemical fertilizer.BroadlyBio-feritlizerIt is referred to as to make a living
Organic fertilizer, is both, containing the nutrient needed for crop, to contain againMicroorganismProduct, be biological, organic and inorganic combination
Body it can replace chemical fertilizer, there is provided cropsGrowthAll kinds of nutrients needed for development.
At present, domestic Liquid Fertilizer production, the actually Water soluble fertilizer of microbial bacterial agent.And generally fermented using solid dung
Tank method produces biological organic fertilizer, is under manual control, under certain temperature, humidity, carbon-nitrogen ratio and ventilation condition, to utilize
The fermentation of the microorganisms such as the widely distributed bacterium of nature, actinomyces, fungi, artificially promotes biodegradable organic
Thing to stabilization humus biochemical conversion microbiological process, i.e., it has often been said that organic fertilizer digest process.Modernization compost
Fermentation tank technique is substantially aerobic compost because aerobic compost has, temperature is high, substrate degradation than more thoroughly, composting week
Phase is short, peculiar smell is small, can on a large scale use mechanical treatment the advantages of.The microbial manure that China has obtained state approval registration has
More than 100 plant, national microbial manure enterprise sum about 950,10,000,000 tons of annual production, and annual value of production is up to 15,000,000,000 yuan.But due to
Science research input wretched insufficiency, technical force relatively lags behind in the market demand, causes some quasi-microorganism fertilizer product fermenting and producings
The quality monitoring setup measures of process are unreasonable, cannot control to product stability and crop field application effect." recurrence has agricultural
Machine fertilizer ", still shoulders heavy responsibilities.Although China is directed toBio-feritlizer（Microbial manure）Research and development, product quality, popularization and application
Etc. aspect pay much attention to, supervision aspect carry out production permit system simultaneously audited and provided by the Ministry of Agriculture.But withBio-feritlizer's
Development, many places are occurred in that without production when bio-feritlizer is producedLicensing, production technology fall behind, with not by identifyBacterium KindProduced, the phenomenon such as effective bacterial content is low, miscellaneous bacteria is exceeded.Also some producers existBio-feritlizerIt is middle to add an inappropriate amount of chemical fertilizer
Or other additives, make that the nitrogen of chemical fertilizer in fertilizer, phosphorus, potassium content are too high to cause effective bacterium dead, and lose the effect of bio-fertilizer.
The content of the invention
The present invention isolates more than three plants by soil-plant-atmosphere system research beneficial microbe from rhizosphere soil
Beneficial microbe microbial inoculum, it is therefore an objective to by generation promotes the growth of microorganism environment of plant growth and suppresses or subtract naturally in soil
Rare opportunistic pathogen of the falling ill three plant bacterium marginal to the infringement of plant, in self-control reactor equipment in preferred ambient temperature
Under conditions of oxygen supply, by suitable nutrient and C:N matrix（Horsehit or cow dung）Culture, generation liquid bio fertilizer.
The purpose of the present invention is achieved through the following technical solutions：
The present invention provide two plants of soil beneficial bacteriums, deposit number be JF-1 and JF-2, its gene order such as SEQ ID No.1 with
Shown in SEQ ID No.2.
The present invention provides three plants of soil beneficial bacteriums is used to prepare the ferment agent of liquid bio-fertilizer, JF-1 therein, JF-2, bacterium
Viable count is 109~1010CFU/ml。
After the beneficial microbial inoculum of three plants of soil of the invention is added with the matrix of cow dung or horsehit, existed by making reactor equipment by oneself
The viable count of JF-1 and JF-2 is respectively 10 in the liquid bio fertilizer generated under conditions of preferred ambient temperature and oxygen supply7~108
CFU/ml。
The fermentation medium cow dung of JF-1 and JF-2 or horsehit are consisted of the following composition：
Cow dung, horsehit nutritive element content（Ordinary circumstance）
Cow dung or the C of horsehit:N is 30-50:1 is optimum as the culture medium of JF-1 and JF-2.
The environmental condition of the fermentation of JF-1 and JF-2 bacterium is in the present invention：Self-control reactor inner cylinder culture medium（Cow dung or horse
Excrement）Volume 0.5m3, outer barrel water volume 1.5m3;Yeasting temperature is 30-40 DEG C;Fermentation time is 12-30h；Initial pH value is
6.5-7.5；Initial inoculum is 1-3%, throughput 40.0L (L/min）.
Optimize technique：35 DEG C of yeasting temperature；PH value is 7.0；Initial inoculum is 2%；Throughput 40.0L (L/
min）；Fermentation time 27h（Stationary phase）.
Beneficial effect
The liquid bio fertilizer of present invention generation, promotes Azobacter to be grown to serve as stronger fixed nitrogen system after being manured into soil；
The present invention is conducive to discharging the nutrient fungus grown in indissoluble mineral；
The present invention is conducive to improving the resistance fungus grown of plant；
The present invention is conducive to degradation of contaminant, reduces toxicity fungus grown；
The present invention may advantageously facilitate the formation fungus grown of detritus acid；
The present invention is conducive to producing plant hormone fungus grown；
The present invention is conducive to biological and ecological methods to prevent plant disease, pests, and erosion fungus grown.
Three plants of soil beneficial bacterium JF-1, JF-2, JF-3 of the invention are processed cow dung or horsehit as leavening,
Increase effective absorption of the plant to nutrient in cow dung or horsehit.
Brief description of the drawings
Fig. 1 is liquid bio fertilizer experimental plot and compares the compares figure after field Chinese cabbage is sowed 10 days；It is liquid bio above figure
Fertile experimental plot, is control field below figure
Specific embodiment
The preparation technology of biological liquid fertilizer of the invention and application are described in detail by following examples, the explanation
It is descriptive, is not intended to limit protection scope of the present invention.
The preparation of the high density fermentation and liquid compound micro-ecological preparation of 1 two kinds of bacterium of embodiment
1st, the preparation of Pseudomonas stutzeri high density fermentation liquid：1）Flat board culture rejuvenation：Pseudomonas stutzeri strain is inoculated in
On LB plating mediums, 18h is cultivated in 30 DEG C, make Pseudomonas stutzeri rejuvenation, and form single bacterium colony, picking single bacterium falls within inoculation
On culture medium, 30 DEG C of culture 24h；2) preparation of first order seed：By step 1）The Pseudomonas stutzeri strain transfer eggplant of culture
On bottle LB slant mediums, 30 DEG C of culture 16h make, in late log phase, to obtain first order seed；3）The preparation of secondary seed：Will step
Rapid 2）The first order seed of preparation is made bacteria suspension with sterilized water, is inoculated into the 100L seeding tanks equipped with 60L LB seed culture mediums
In, 30 DEG C of temperature, rotating speed 250rpm, tank pressure 0.05MPA, ventilating ratio：1：0.6, culture 146 obtains secondary seed solution.4）Amur is false
The preparation of unit cell fermented liquid：By step 3）The secondary seed solution of preparation is inoculated into 10m according to 5% inoculum concentration3Fermentation medium
Fermentation tank in, 30 DEG C of temperature, rotating speed 200rpm, tank pressure 0.05Mpa, ventilating ratio：1：0.6, cultivate 16h, viable count be 3.2 ×
109The Pseudomonas stutzeri zymotic fluid of CFU/ml；Described fermentation medium is：Tryptone 10g, yeast extract 5g, chlorine
Change sodium 10g, deionized water 1000ml, pH are 7.0.2nd, the preparation of bacillus amyloliquefaciens high density fermentation liquid：1）Flat board culture
Rejuvenation：Bacillus amyloliquefaciens strain is inoculated on BPY plating mediums, 24h is cultivated in 30 DEG C, make bacillus amyloliquefaciens
Rejuvenation, and single bacterium colony is formed, picking single bacterium is fallen within inoculation medium, 30 DEG C of culture 24h；2) preparation of first order seed：Will step
Rapid 1）On the bacillus amyloliquefaciens strain transfer eggplant bottle BPY slant mediums of culture, 37 DEG C of culture 24h make in logarithm
In the later stage, obtain first order seed；3）The preparation of secondary seed：By step 2）The first order seed of preparation is made bacteria suspension with sterilized water, connects
Plant in the 100L seeding tanks equipped with 60L BPY seed culture mediums, 30 DEG C of temperature, rotating speed 300rpm, tank pressure 0.05MPA, ventilation
Than：1：1 culture 14h, obtains secondary seed solution.4）The preparation of bacillus amyloliquefaciens zymotic fluid：By step 3）The two grades of kinds for preparing
Sub- liquid is inoculated into the 1m of 600L fermentation mediums by 5% inoculum concentration3In fermentation tank, 30 DEG C of temperature, rotating speed 300rpm, tank pressure
0.05Mpa, ventilating ratio 1：1,16h is cultivated, gemma production rate more than 90% is obtained, viable count is 5 × 109The withered grass gemma of cfu/ml
Bacillus fermentation liquid；Described fermentation medium is（Mass percent）：Sucrose 1.0%, peptone 1.0%, yeast extract 0.5%,
Ammonium sulfate 0.2%, magnesium sulfate 0.04%；Manganese sulfate 0.08%, defoamer 0.06%, pH is 6.8.3rd, biological liquid fertilizer leavening
Preparation：The zymotic fluid that 1 and 2 are obtained is by 1：PH is adjusted after 1 ratio is well mixed to 7.0 or so, carry out it is filling, every bottle
1000mL, plus 10% sodium chloride, are obtained bacterium number probably 4.0 × 109The liquid fertilizer leavening of CFU/ml.
The biological liquid fertilizer preparation technology of embodiment 2
Fermenting agent obtained in example 1 is added in cow dung or horsehit reactor according to 1% inoculum concentration, initial pH7.0, sent out
Ferment temperature control is in 30~37 DEG C, throughput 40.0L (L/min）, fermentation time be 12~30h, after fermentation ends, JF-1 and
The viable count of JF-2 is 107~108CFU/ml.Liquid fertilizer color dark-brown or color of soy sauce, solid content about 3~5%.Liquid
Fertilizer is packed using plastic products, packing specification 1000ml/ bottles.
Self-control reactor inner cylinder culture medium（Cow dung or horsehit）Volume 0.5m3, outer barrel water volume 1.5m3, with temperature control
System and compressed air air-breather.
The biological liquid fertilizer application test of embodiment 4
In Tongzhou international seed industry garden and Shunyi institute of agricultural sciences Great Sun Ge Village Zhao Jia valleys base, improvement of soil fertility proving ground is set：Grainfield
It is summer corn and winter wheat crop rotation, open country field of vegetables is Chinese cabbage and eggplant crop rotation.Crop varieties：Winter wheat agricultural university 211, summer corn capital
Single 28, Chinese cabbage is new No. 3, eggplant is black treasured.Two places grainfield, field of vegetables experimental cultivation area are 0.5 mu.After using 10 days,
Compared with control group, long-term cropping bud ratio improves 10%, reaches 90% bud ratio in biological liquid fertilizer experimental plot；Plant height ratio
Control group 3~5cm high, reaches 8~10cm, and plant growth regularity is significantly improved；Chlorophyll content experimental group ratio in the middle of cauline leaf
Control group is high by 10%.Experimental plot is with to compare field growing state as shown in Figure 1.
Claims (8)
1. a kind of biological liquid fertilizer that prepared by utilization cow dung or horsehit fermentation, its feature includes Pseudomonas stutzeri
（Pseudomonas stutzeri JF-1）With bacillus amyloliquefaciens (Bacillus amyloliquefaciens JF-2).
2. Pseudomonas stutzeri 16sRNA gene orders such as SEQ ID No as described in right 1:Shown in 1, bacillus amyloliquefaciens
16sRNA gene orders such as SEQ ID No:Shown in 2.
3. two plants of microorganisms, for cow dung or the liquid fermentation of horsehit, are prepared comprising two kinds of liquid of microorganisms as described in right 1
Fertilizer.
4. application of the two plants of soil beneficial bacteriums described in claim 3 in liquid bio-fertilizer is prepared, it is characterised in that by base
Matter（Horsehit or cow dung）Nutrient and C:N determines the microbial bacterial agent of optimum compound proportion（JF-1/JF-2).
5. self-control reactor equipment generates product under conditions of preferred ambient temperature and oxygen supply.
6. as described in right 1 and right 3, two kinds of microbial liquids are in cow dung or horsehit fermentation process according to 1:1 mode adds
Plus.
7. as described in right 1 and right 5, JF-1 fermentation density is 109~1010CFU/ml, JF-2 fermentation density are 109~
1010CFU/ml。
8. growing environment and the suppression of the microorganism and its enzyme of generation promotion plant growth naturally is applied in soil as described in right 3
System reduces harmful infringement of the pathogen to plant, while reducing the nutrient loss that oxidation is caused.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110172413A (en) * | 2019-03-21 | 2019-08-27 | 内蒙古和美科盛生物技术有限公司 | A kind of compound probiotic agent and its application being used to prepare liquid bio fertilizer |
| CN111233526A (en) * | 2020-01-21 | 2020-06-05 | 南京工业大学 | Bio-based water-soluble fertilizer fermented by pentamethylene diamine fermentation tail liquid and preparation method and application thereof |
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| CN104557235A (en) * | 2013-10-11 | 2015-04-29 | 朱晓琳 | Cattle manure fertilizer and preparing method thereof |
| CN103667122A (en) * | 2013-11-27 | 2014-03-26 | 北京龙科方舟生物工程技术有限公司 | Compound bacterium and application thereof |
| CN103992159A (en) * | 2014-03-25 | 2014-08-20 | 南京农业大学 | Full-element compound microbial fertilizer and preparation method and application thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110172413A (en) * | 2019-03-21 | 2019-08-27 | 内蒙古和美科盛生物技术有限公司 | A kind of compound probiotic agent and its application being used to prepare liquid bio fertilizer |
| CN111233526A (en) * | 2020-01-21 | 2020-06-05 | 南京工业大学 | Bio-based water-soluble fertilizer fermented by pentamethylene diamine fermentation tail liquid and preparation method and application thereof |
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Application publication date: 20170531 |