CN107794231A - A kind of feature bacterial strain, fermentation process, production method and application - Google Patents
A kind of feature bacterial strain, fermentation process, production method and application Download PDFInfo
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- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
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Abstract
The invention provides a kind of feature bacterial strain, is bacillus amyloliquefaciens (Bacillus amylolique faciens) LVLE15, preservation date is on June 20th, 2016, and deposit number is CGMCC No.12640.Present invention also offers the method for bacillus amyloliquefaciens production liquid composite microorganism fertilizer and solid composite microbe fertilizer, bacillus amyloliquefaciens fermenting and producing, the zymotic fluid by 0.1 3% mass ratio be added to liquid organic starting material or processing after SOLID ORGANIC raw material in.The bacillus amyloliquefaciens are additionally provided especially to suppress cucumber early blight in suppression crop disease and suppress the application on gray mold of cucumber.The present invention bacillus amyloliquefaciens and its obtained zymotic fluid, fertilizer, there is effective biocontrol effect, to cucumber early blight and gray mold of cucumber prevention effect it is further preferred that.
Description
Technical field
The present invention relates to a kind of Antagonistic Fungi that can prevent and kill off various crop disease --- biocontrol water bacillus amyloliquefaciens, simultaneously
It is related to production method of its related fertilizer etc..
Background technology
China's controlling disease substantially uses chemical pesticide at present, and chemical pesticide control is with its strong toxicity, prevention effect
Well, cost is low and is generally welcome by peasant.The agricultural chemicals that China produces every year kind about more than 250, process preparation kind more than 500, active compound life
More than 80 ten thousand tons of production, occupies the 2nd, the world.Agricultural chemicals annual consumption is ten thousand tons of 80-100, is occupied first place in the world.But largely use chemical agriculture
Medicine, first, causing pollution by pesticides to environment, second, bringing Practice for Pesticide Residue in Agricultural Products problem, prestige is caused to human health
The side of body.To reduce the use of chemical pesticide, cultural control in recent years, biological green preventing and treating are at home and abroad progressively risen.
Exploitation biological pesticide and the microbial-bacterial fertilizer with biological and ecological methods to prevent plant disease, pests, and erosion characteristic have become a kind of international trend.The world is each
State take practical steps reduce chemical pesticide use, as U.S. EPA 90 years with regard to announce cancel 91 kinds of chemical pesticides
Registration.The development and application of biological pesticide is widely paid close attention to and paid attention to, and it is new to have turned into agricultural chemicals career field in the world at present
Research and development focus.Biological control foreign countries on soil-borne disease begin one's study the twenties, as nineteen twenty-one C.Hartley reports can be with
With ten several common soil earth saprophytic fungus and the rotten mould root rot of bacterium preventing and treating sapling.40th, utilization fifties Antagonism is put
Line bacterium and caused antibiotic, turn to the sixties and find other antagonistic microbes and biological and ecological methods to prevent plant disease, pests, and erosion technical research.Report in the world
Bio-control factors applied to soil-borne disease are a lot, including fungi, bacterium, viral phage and defence line bacterium etc., which part product
For merchandized handling, and more and more important effect is played in integrated pest prevention and control and agricultural sustainable development.Mesh
Before, the output value of biological pesticide is all increasing in 10-20% speed every year in the world.Reduce the use of chemical synthetic pesticide, hair
Open up biological pesticide turn into a kind of trend and direction it is contemplated that.
Based on farm antibiotics, mainstream product includes jinggangmycin, blasticidin-S, resisted more for the exploitation of domestic biological pesticide
Mycin, kasugarnycin, valida and mildiomycin etc. are used for preventing and treating the use such as fungal disease, streptomysin, penicillin and terramycin
To prevent and treat bacteriosis；The biological living agricultural chemicals succeeded in developing at present also has sub- treasured, power treasured, waxy Bacillus, yield increasing fungus (to plant
Thing growth regulator), K84, E26, the bacteria preparation such as Pseudomonas alba and fluorescence vacation monospore, the preventing and treating for soil-borne disease.Its
The anti-crown gall microbial inoculum K84 registered registrations at present of middle China Agricultural University's exploitation, product are that 2,000,000 cfu/ milliliters radiate soil
Bacillus wet-milling, to root knot effects such as kernel approaches (peach, Lee, cherry, apricot etc.) up to 100%；To pomaceous fruit and forest, flowers crown gall
Disease has better effects.The gemma bacillus agent also registered registration at present of Yunnan agricultural university exploitation, product is 1,000,000,000/g bacillus
WP (hundred is anti-), has good preventive effect to rice sheath blight disease.Other parts scientific achievement also be applied to production practices (especially without
Public hazards food production base), and achieve good ecology, social and economic benefit.
The content of the invention
Therefore, the technical problem to be solved in the present invention is to provide a kind of bacterial strain with good biocontrol effect, especially
There is the bacterial strain of good biocontrol effect to early blight and suppression gray mold.The invention solves another technical problem, be
Bio-feritlizer is produced using the bacterial strain, quality meets the Ministry of Agriculture《Composite microbiological fertilizer》(NY/T798-2015) requirement.
The technical scheme is that a kind of feature bacterial strain, the bacterial strain is bacillus amyloliquefaciens (Bacillus
Amylolique faciens) LVLE15, China Committee for Culture Collection of Microorganisms's common micro-organisms center is deposited in, is protected
It is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 to hide address；Preservation date is on June 20th, 2016, and deposit number is CGMCC
No.12640。
Main Biological is：In Antagonistic Fungi bacillus amyloliquefaciens LVLE15 zymotic fluids living bacteria count up to 15,000,000,000/
Ml, gemma rate 93%, living bacteria count reaches 1.5*10 in the bio-feritlizer produced with LVLE158Individual/more than g.
Thalline is shaft-like, size 0.3-2.2 × 1.2-7.0 microns.Gram-positive, optimum growth temperature 25-35
DEG C, pH is neutral.
The fermentation culture method of above-mentioned feature bacterial strain, the hair of the bacillus amyloliquefaciens LVLE15 liquid fermentations production
Ferment condition is：PH is 6.5-8.0, and cultivation temperature is 30-45 DEG C, originates rotating speed 120-200rpm, throughput 0.5-1vvm, training
Support 35-45 hours, for fermentation middle and later periods row into gemma, gemma rate is up to more than 93%, and the living bacteria count in zymotic fluid is 15,000,000,000
Individual/more than ml.
Strain fermentation culture medium prescription be corn flour 20-30g, beancake powder 20-30g, urea 0.2-2g, sucrose 1-5g,
K2HPO40.5-2.5g、MgSO40.1-0.6g、MnSO40.05-0.5g, bubble enemy 0.1-0.5g, pH 7.2, are then diluted to
1000ml.Living bacteria count is 1.49 × 10 in microbial inoculum10It is more than individual/milliliter.
Present invention also offers a kind of method that composite microbiological fertilizer is produced with bacillus amyloliquefaciens, by above-mentioned Xie Dian
Afnyloliquefaciens fermenting and producing, gained zymotic fluid are added in liquid organic starting material by 0.1-3% mass ratio, are obtained liquid and are answered
Close microbial manure.
Preferably, the content of organic matter of the liquid organic starting material is more than 30%.
Further, the liquid organic starting material preferably is selected from yeast waste liquor or amino acid waste liquid.
Present invention also offers the liquid composite microorganism fertilizer that above-mentioned method obtains, the liquid composite microbial fertilizer
Material includes bacillus amyloliquefaciens zymotic fluid and liquid organic starting material, and the bacillus amyloliquefaciens zymotic fluid accounts for liquid bio fertilizer
The 0.1-3% of gross weight.
The present invention also provides a kind of method that composite microbiological fertilizer is produced with bacillus amyloliquefaciens microbial inoculum, and solid is had
The pH controls of machine raw material are in 5.0-8.0, crushing；By above-mentioned bacillus amyloliquefaciens fermenting and producing, gained zymotic fluid is situated between with absorption
Solid fungicide is obtained after matter absorption, solid fungicide is added in solid, powdery Organic Ingredients according to 2%-10% mass ratioes, obtained
To solid composite microbe fertilizer.The medium of absorption can be turf or powdered rice hulls.
Preferably, the content of organic matter of the SOLID ORGANIC raw material is more than 30%.
Further, the SOLID ORGANIC raw material is selected from the hydrolysis residue or edible fat production process of amino acids production process
The dregs of fat.
It is preferred that the main component of several raw materials：
Yeast waste liquor：Purchased from Angel Yeast Co., Ltd.Main component is as follows：
Table 1：Yeast concentrate main nutrient composition table
Production process of the hydrolysis residue from amino acid such as Gourmet Powder Factory, lysine factories.The base of hydrolysis residue after treatment
This character is as follows：
Table 2：The Elemental characters of hydrolysis residue (powdery) after processing
The dregs of fat come from Shanghai Li Chang environment Co., Ltd, and main component is as follows：
Table 3：Dregs of fat main nutrient composition table
Present invention also offers the composite microbiological fertilizer that the above method is prepared, the composite microbiological fertilizer includes
Solid bacillus amyloliquefaciens microbial inoculum and SOLID ORGANIC raw material, the solid bacillus amyloliquefaciens microbial inoculum account for composite microbe fertilizer
The 2%-10% of the gross weight of material.
Preferably, the one or more of npk fertilizer can also be added in the composite microbiological fertilizer.Such as
The inorganic element content of itself, N+P in fruit Organic Ingredients2O5+K2O ＜ 8%, that just needs appropriate addition inorganic elements, i.e. nitrogen
Fertilizer, phosphate fertilizer, one kind in potash fertilizer.Preferably N is supplemented from urea, P2O5What is selected is potassium phosphate, K2What O was selected is nitric acid
Potassium.Macroelement fertilizer can also can be added according to soil and crop needs.Trace element fertilizer can also be added as needed
Material.
Especially suppress early blight and/or suppression in suppression crop disease present invention also offers above-mentioned bacillus amyloliquefaciens
Application in gray mold processed.
The bacillus amyloliquefaciens of the present invention can be individually used for suppressing early blight or be individually used for suppressing gray mold, or
Person is used to suppress early blight and gray mold jointly.
Beneficial effects of the present invention
The present invention is a kind of microbial manure and its production method with Biological control function, using rich in organic matter
The raw material of crop fertilizer, then bio-feritlizer is made after being mixed with the zymotic fluid of Antagonistic Fungi, the product with the market
Product is compared and had the following advantages that：
1) Antagonistic Fungi is rich in fertilizer product, its biocontrol effect highly significant, the content of the bacterial strain is up to 100,000,000 in fertilizer
Individual/more than g/ml, dominant microflora can be formed after being manured into soil, and be survived in soil.
2) the preferred content of organic matter of Organic Ingredients that the present invention selects is higher, and nutrient content is higher, is not required to substantially
Add inorganic nutrients.
3) product of the present invention has obvious inhibition to cucumber early blight and gray mold, in treadmill test, preventive effect point
74.85% and 80.64% are not reached.
4) biocontrol water bio-feritlizer has diseases prevention and fertilizer efficiency effect concurrently after applying, and can improve crop resistance, and to fertilizer
Absorption and utilization rate, keep plant growth healthy and strong, can also promote the breeding of biocontrol microorganisms to improve crop disease-resistant ability in turn, and
Help to form good agricultural eco-system.
5) pesticide dosage can be reduced while fertilizer amount is reduced, residues of pesticides and pollution are reduced, for sustainable agriculture
The raising of the development and people's living standard of industry and pollution-free food production plays an important roll.
Embodiment
Strain isolation process
1. soil sample gathers and processing
Serious field occurs from the purpleflower violet herb growing area epidemic disease such as Shanghai, Jiashan, Pinghu, using five point sampling methods from growth
Healthy cucumber rhizosphere takes soil sample.Top layer 5cm soil layers are scalped during sampling, fresh soil sample 500g is taken, is preserved with freshness protection package.Weigh soil
Sample 10g is added in the triangular flask equipped with 90mL sterilized waters (built-in bead is some), 30 DEG C, 120r/min vibration 30min, is stood
After obtain soil supension.
2. the screening of biocontrol bacteria
Soil supension is diluted to 10-4、10-5、10-6Concentration, the μ L of soil supension 100 for drawing different dilution factors are coated on
On NA culture mediums, it is placed in 30 DEG C and cultivates 48 hours, the different bacterium single bacterium colony such as picking color, form is simultaneously purified, preserved.Using
Bacteriostatic activity of the bacterium that flat board face-off method measure is separated to powdery mildew of cucumber.Powdery mildew of cucumber is activated on PDA plate,
Beaten with card punch in colony edge and take diameter 6mm fungus blocks, be inoculated into PDA plate center.Apart from powdery mildew of cucumber after cultivating 1 day
4 docking points inoculation tested bacteria at fungus block 2cm, 28 DEG C of cultures, observes antibacterial result, each processing sets 3 weights after 3-5 days
It is multiple.Choose the bacterial strain and numbering LVLE-15 of good antimicrobial effect.
3. the identification of antagonistic strain
3.1 bacterial strain bio-chemical characteristics：The bacterial strain of the fungistatic effect screened is subjected to colony morphological observation, microscope shape
State observation, Gram's staining, spore staining, hydrolysis starch experiment, methyl red test (M.R), V.P experiments, citrate test,
The Main Morphologies such as catalase reaction test, gelatin liquefaction test, mobility test and physiological and biochemical index identification.Preliminary Identification
LVLE-15 is bacillus amyloliquefaciens category.
The 16S rDNA sequence analyses identification of 3.2 bacterial strains
By carrying out 16S rDNA sequencings and comparative analysis to bacterium, the results showed that, LVLE-15 16S rDNA and Xie Dian
The 16S rDNA of afnyloliquefaciens have a homology of height, and homology is up to 99%.What comparison result obtained with cell classification method
Qualification result is consistent.
Embodiment 1-1
1st, microbial inoculum：Strain fermentation culture medium prescription be corn flour 25g, beancake powder 25g, urea 1g, sucrose 3g,
K2HPO41.2g、MgSO4 0.3g、MnSO40.2nd, bubble enemy 0.3g, pH 7.2, water is added to 1000ml.In microbial inoculum effectively
Viable count is 1.49 × 1010Individual/milliliter.
Bacillus amyloliquefaciens LVLE15 liquid fermentations production fermentation condition be：PH is 6.5-8.0, and cultivation temperature is
30-45 DEG C, originate rotating speed 120-200rpm, throughput 0.5-1vvm, cultivate 35-45 hours, fermentation middle and later periods row into gemma,
Gemma rate is up to more than 93%, and the living bacteria count in zymotic fluid is in 15,000,000,000/more than ml.
2nd, composition of raw materials is matched somebody with somebody：Added according to formula as below.Trace element is borax and chelated zinc, ratio 1.5:3.5；Protect
Shield agent is sodium benzoate and molasses, ratio 1.5:2.
The liquid composite microorganism fertilizer raw material of table 4 addition formula
3rd, inorganic nutrients are mixed with 40% (gross mass) water；Protective agent is added in concentrate, again will after stirring
Bacterium solution adds；Finally mixed by the inorganic nutrients fully dissolved and containing bacterium concentrate, and add remaining moisture, it is compound that liquid is made
Microbial manure.Finished product fertilizer all technical is as follows.
The liquid composite microorganism fertilizer the key technical indexes of table 5
4th, liquid composite microorganism fertilizer is dispensed through liquid filling device, seals, is put in storage.
Embodiment 1-2
1st, the preparation of solid microbial：Bacterium solution turf after biocontrol water bacillus amyloliquefaciens LVLE15 is fermented
By bacterium solution：Turf=1：5 ratio is adsorbed, and the living bacteria count amount of composite bacteria agent is up to 1.2 × 10 after mixing9It is individual/gram with
On, water tariff collection is within 30%.
2nd, sample.Caused hydrolysis residue during amino acids production, it is derived from Henan "Lotus" gourmet powder company limited by shares；
3rd, pH is nursed one's health：Break into bulk hydrolysis residue into pieces powder or little particle, hydrolysis residue is pressed with powdered lime：Lime=4：1
Mixing, 8-12 hours are stacked after being crushed with chain crusher, allow it fully to react；
4th, turn upside down：With forklift or artificial turning, reacted temperature of charge cooling is allowed to reach normal temperature；
5th, crush：The hydrolysis residue for being cooled to normal temperature is measured, fundamental property is shown in Table 6, content of beary metal well below
The professional standard of biological organic fertilizer, is not measured；
Table 6：The Elemental characters of hydrolysis residue after processing
6th, dispensing.Added in mixer and be slowly added to powdery hydrolysis residue, and preparation is added by weight 6% ratio
Good complex micro organism fungicide.
7th, mix.Mixer stirs 3-5 minutes, material is sufficiently mixed uniformly.
8th, crush.The fineness of material is set to reach 60-80 mesh by pulverizer.
9th, pack, measure, storage.
Embodiment 1-3
1st, the preparation of solid microbial：Bacterium solution turf after biocontrol water bacillus amyloliquefaciens LVLE15 is fermented
By bacterium solution：Turf=1：5 ratio is adsorbed, and the living bacteria count amount of composite bacteria agent is up to 1.2 × 10 after mixing9It is individual/gram with
On, water tariff collection is within 30%.
2nd, sample.The dregs of fat caused by absorption, are derived from Shanghai Li Chang environment Co., Ltd in salad oil production process；
3rd, pH is nursed one's health：Break into the bulk dregs of fat into pieces powder or little particle, hydrolysis residue is pressed with powdered lime：Lime=4：1 is mixed
Close, 8-12 hours are stacked after being crushed with chain crusher, allow it fully to react；
4th, turn upside down：With forklift or artificial turning, reacted temperature of charge cooling is allowed to reach normal temperature；
5th, crush：The dregs of fat for being cooled to normal temperature are measured, fundamental property is shown in Table 7, and content of beary metal is well below life
The professional standard of organic fertilizer, is not measured；
Table 7：The Elemental characters of the dregs of fat after processing
6th, dispensing.Added in mixer and be slowly added to the powdery dregs of fat, and prepared by weight 6% ratio addition
Complex micro organism fungicide.
7th, mix.Mixer stirs 3-5 minutes, material is sufficiently mixed uniformly.
8th, crush.The fineness of material is set to reach 60-80 mesh by pulverizer.
9th, pack, measure, storage.
Table 8：The bio-organic fertilizer product quality measurements of different material production
As can be seen from the above table, in terms of bacteriostasis rate, inside solid material, the dregs of fat are more suitable for Xie Dian as raw material
Afnyloliquefaciens.
Test example
1 experimental strain
1.1 bacillus
LVLE15：Bacillus amyloliquefaciens
1.2 disease fungus
Early blight, leaf mold, anthracnose, gray mold
2. experimental method
2.1 bacterial strains activate
It is standby after the activation of NA flat boards to screen bacterium, disease fungus PDA plate is standby after overactivation.
The preparation of 2.2 gemma filtered fluids
Gemma is inoculated with into NB culture mediums 28 DEG C, 200rpm culture 48h, normal temperature, 5000g centrifugations 5min obtain supernatant.Pass through again
It is standby to cross 0.22um filter filtration sterilizations.
It is prepared by 2.3 detection flat boards
With 1:10 ratios addition filtered fluid mixes pour plate into 50 DEG C of 10/9 times of concentration PDA culture mediums, per ware 20ml.
With 1:10 ratios addition NB culture mediums substitute fermentation filtrate as control.
2.4 disease funguses are inoculated with
Beaten with card punch (5mm) and take pathogen dish to be placed in corresponding flat board center, pathogen is connect on flat board as control to compare,
Experiment sets 3 repetitions.
2.5 culture observations
Culture dish is placed at 28 DEG C and cultivated, it is straight in the 3rd day, observation detection in the 5th day, crossing method measurement cause of disease bacterium colony
Footpath.Average value is calculated, calculates bacteriostasis rate.
Bacteriostasis rate (%)=[(control average diameter-experimental group average diameter)/control average diameter] × 100%
3 experimental result tables
Table 9：Histamine result of the bacillus amyloliquefaciens fermentation filtrate to various germs
From above test effect can be seen that bacillus amyloliquefaciens LVLE15 used in the present invention to cucumber early blight and
Gray mold is with obviously preventive effect, preventive effect are up to 74.85% and 80.64%.
Claims (10)
- A kind of 1. feature bacterial strain, it is characterised in that：The bacterial strain is bacillus amyloliquefaciens (Bacillus amylolique Faciens) LVLE15, is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preservation address is north Jing Shi；Preservation date is on June 20th, 2016, and deposit number is CGMCC No.12640.
- 2. the fermentation culture method of the feature bacterial strain described in claim 1, it is characterised in that：The bacillus amyloliquefaciens LVLE15 liquid fermentations production fermentation condition be：PH is 6.5-8.0, and cultivation temperature is 30-45 DEG C, starting rotating speed 120- 200rpm, throughput 0.5-1vvm, to cultivate 35-45 hours, fermentation middle and later periods row is up to more than 93% into gemma, gemma rate, Living bacteria count in zymotic fluid is in 15,000,000,000/more than ml.
- A kind of 3. method that liquid composite microorganism fertilizer is produced with biocontrol water bacillus amyloliquefaciens, it is characterised in that：Will power Profit requires the 1 bacillus amyloliquefaciens fermenting and producing, and it is organic that obtained zymotic fluid by 0.1-3% mass ratio is added to liquid In raw material, liquid composite microorganism fertilizer is obtained.
- 4. the method according to claim 3 that liquid composite microorganism fertilizer is produced with biocontrol water bacillus amyloliquefaciens, It is characterized in that：The liquid organic starting material is selected from yeast waste liquor or amino acid waste liquid.
- 5. the liquid composite microorganism fertilizer that the method described in claim 3 obtains, it is characterised in that：The liquid composite microbial Thing fertilizer includes bacillus amyloliquefaciens zymotic fluid and liquid organic starting material, and the bacillus amyloliquefaciens zymotic fluid accounts for liquid life The 0.1-3% of thing fertilizer gross weight.
- A kind of 6. method that composite microbiological fertilizer is produced with biocontrol water bacillus amyloliquefaciens microbial inoculum, it is characterised in that：Will be solid The pH controls of body Organic Ingredients are in 5.0-8.0, crushing；By bacillus amyloliquefaciens fermenting and producing described in claim 1, gained hair Zymotic fluid obtains solid fungicide after being adsorbed with adsorbing medium, and solid fungicide is added into solid, powdery according to 2%-10% mass ratioes has In machine raw material, solid composite microbe fertilizer is obtained.
- 7. the method according to claim 6 that composite microbiological fertilizer is produced with biocontrol water bacillus amyloliquefaciens microbial inoculum, It is characterized in that：The content of organic matter of the SOLID ORGANIC raw material is more than 30%；The SOLID ORGANIC raw material is selected from amino acid The hydrolysis residue of production process or the dregs of fat of edible fat production process.
- 8. the composite microbiological fertilizer that claim 6 methods described is prepared, it is characterised in that：The composite microbiological fertilizer Including solid bacillus amyloliquefaciens microbial inoculum and SOLID ORGANIC raw material, the solid bacillus amyloliquefaciens microbial inoculum accounts for composite microbial The 2%-10% of the gross weight of thing fertilizer.
- 9. the side that composite microbiological fertilizer is produced with biocontrol water bacillus amyloliquefaciens microbial inoculum according to claim 3 or 6 Method, it is characterised in that：The one or more of N,P,K fertilizers can also be added in the composite microbiological fertilizer.
- 10. feature bacterial strain described in claim 1 especially suppresses early blight in suppression crop disease and/or suppressed in gray mold Application.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610756981.1A CN107794231B (en) | 2016-08-29 | 2016-08-29 | Functional strain, fermentation method, production method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610756981.1A CN107794231B (en) | 2016-08-29 | 2016-08-29 | Functional strain, fermentation method, production method and application |
Publications (2)
| Publication Number | Publication Date |
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| CN107794231A true CN107794231A (en) | 2018-03-13 |
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| CN108424856A (en) * | 2018-03-22 | 2018-08-21 | 河北邢化生态环境工程有限公司 | A method of producing agricultural microbial agent using maize germ grease |
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| CN108424856A (en) * | 2018-03-22 | 2018-08-21 | 河北邢化生态环境工程有限公司 | A method of producing agricultural microbial agent using maize germ grease |
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