CN107779413A - One plant of biocontrol water bacillus subtilis, production method, liquid fertilizer and application - Google Patents
One plant of biocontrol water bacillus subtilis, production method, liquid fertilizer and application Download PDFInfo
- Publication number
- CN107779413A CN107779413A CN201610756959.7A CN201610756959A CN107779413A CN 107779413 A CN107779413 A CN 107779413A CN 201610756959 A CN201610756959 A CN 201610756959A CN 107779413 A CN107779413 A CN 107779413A
- Authority
- CN
- China
- Prior art keywords
- bacillus subtilis
- fertilizer
- liquid
- solid
- organic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
- C05F5/002—Solid waste from mechanical processing of material, e.g. seed coats, olive pits, almond shells, fruit residue, rice hulls
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
- C05F5/006—Waste from chemical processing of material, e.g. diestillation, roasting, cooking
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- 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
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- 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
- C05G5/00—Fertilisers characterised by their form
- C05G5/20—Liquid fertilisers
- C05G5/23—Solutions
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Pest Control & Pesticides (AREA)
- Environmental & Geological Engineering (AREA)
- Botany (AREA)
- Medicinal Chemistry (AREA)
- Virology (AREA)
- Mechanical Engineering (AREA)
- Fertilizers (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides the bacillus subtilis that one plant has suppression disease activity, the bacillus subtilis with suppression disease activity is bacillus subtilis (Bacillus subtilis) LVLE14, preservation date is on June 20th, 2016, and deposit number is CGMCC No.12639.Present invention also offers the method for bacillus subtilis production liquid bio fertilizer, fermentation of bacillus subtilis production, the zymotic fluid to be added in liquid organic starting material by 0.1 3% mass ratio, obtains liquid bio fertilizer.The production method of bio-organic fertilizer is provided simultaneously.The bacillus subtilis is additionally provided especially to suppress powdery mildew of cucumber in suppression crop disease and suppress the application on gray mold of cucumber.The present invention bacillus subtilis and its obtained zymotic fluid, fertilizer, there is effective biocontrol effect, to powdery mildew of cucumber 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 subtilis and its life
The production method of thing fertilizer.
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 powdery mildew of cucumber.The invention solves another technical problem, be to utilize the bacterium
Strain production bio-feritlizer, quality meet the Ministry of Agriculture《Composite microbiological fertilizer》(NY/T798-2015) requirement.
The technical scheme is that one plant of bacillus subtilis with suppression disease activity, it is characterised in that：It is described to have
The bacillus subtilis of suppression disease activity is bacillus subtilis (Bacillus subtilis) LVLE14, is deposited in the micro- life of China
Thing culture presevation administration committee common micro-organisms center (CGMCC), preservation address are Yard 1, BeiChen xi Road, Chaoyang District, Beijing Cities
No. 3；Preservation date is on June 20th, 2016, and deposit number is CGMCC No.12639.
Main Biological is：In Antagonistic Fungi bacillus subtilis LVLE14 zymotic fluids living bacteria count up to 10,000,000,000/
Ml, gemma rate 95%, living bacteria count reaches 1*10 in the bio-feritlizer produced with LVLE148Individual/more than g.
Gemma 0.6-0.9 × 1.0-1.5 microns, ellipse arrive column, no pod membrane, and thalline does not expand after sporulation.Bacterium colony
Rough surface is opaque, Gram-positive, 30-40 DEG C of optimal temperature, pH6.5-7.5.
The above-mentioned fermentation culture method with the active bacillus subtilis of suppression disease, is the bacillus subtilis LVLE14
Liquid fermentation production fermentation condition be：PH6.0-8.0, cultivation temperature are 20-40 DEG C, originate rotating speed 100-200rpm, ventilation
Measure as 0.5-1vvm, cultivate 35-40 hours, for fermentation middle and later periods row into gemma, gemma rate is up to more than 95%, having in zymotic fluid
Viable count is imitated in 10,000,000,000/more than ml.
Preferably, strain fermentation culture medium prescription：Soluble starch 20-40g, analysis for soybean powder 5-30g, sucrose 1-10g, egg
White peptone 2-8g, dregs of beans 1-6g, K2HPO4 1-5g、MgSO40.2-3g, bubble enemy 0.1-2g, moisturizing is to 1000 milliliters.
Present invention also offers a kind of method that liquid bio fertilizer is produced with bacillus subtilis microbial agent, by above-mentioned withered grass bud
Spore bacillus fermentation is produced, and gained zymotic fluid is added in liquid organic starting material by 0.1-3% mass ratio, obtains liquid bio
Fertilizer.
Preferably, the content of organic matter of the liquid organic starting material is more than 30%.
Preferably, the liquid organic starting material preferably is selected from yeast waste liquor or amino acid waste liquid.
The present invention also provides the liquid bio fertilizer that the above method produces to obtain, and the liquid bio fertilizer includes bacillus subtilis
Fermented liquid and liquid organic starting material, the bacillus subtilis fermentation liquor account for the 0.1-3% of liquid bio fertilizer gross weight.Should
Liquid bio fertilizer may also include some other auxiliary material, such as preservative.
The present invention also provides a kind of production method that solid composite microbe fertilizer is produced with bacillus subtilis microbial agent, will
The pH controls of SOLID ORGANIC raw material are in 5.0-8.0, crushing；Fermentation of bacillus subtilis described in claim 1 is produced, gained hair
Zymotic fluid obtains solid bacillus subtilis microbial agent after being adsorbed with adsorbing medium, solid fungicide is added according to 2%-10% mass ratioes
Into solid, powdery Organic Ingredients, solid composite microbe fertilizer is obtained.
The medium of absorption can be turf or powdered rice hulls.The absorption part by weight of bacterium solution and adsorbing medium is usually 1：2-
5。
Preferably, the content of organic matter of the SOLID ORGANIC raw material is more than 30%.
Above-mentioned bacillus subtilis LVLE14 microbial inoculum, the mixing compared with high organic can be contained with other, such as dregs of beans material
Deng.Preferably, the SOLID ORGANIC raw material is selected from the hydrolysis residue of amino acids production process or the oil of edible fat production process
Slag.
Present invention also offers the biological organic fertilizer that above-mentioned method produces to obtain, it is withered that the biological organic fertilizer includes solid
Careless gemma bacillus agent and SOLID ORGANIC raw material, the solid bacillus subtilis microbial agent account for the gross weight of biological organic fertilizer
2%-10%.
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
Preferably, the one or more of npk fertilizer can also be added in aforesaid liquid bio-fertilizer.Above-mentioned life
Thing fertilizer can also add the one or more of npk fertilizer.Preferably N is supplemented from urea, P2O5What is selected is phosphorus
Sour potassium, K2What O was selected is potassium nitrate.Macroelement fertilizer can also can be added according to soil and crop needs.Can also basis
Need to add trace-element fertilizer.
Especially suppress powdery mildew of cucumber and/or suppression in suppression crop disease present invention also offers bacillus subtilis
Application on gray mold of cucumber.
The bacillus subtilis of the present invention can be individually used for suppressing powdery mildew of cucumber or be individually used for suppressing cucumber ash
Mildew, or for suppressing powdery mildew of cucumber and gray mold of cucumber jointly.
Beneficial effect
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 powdery mildew of cucumber, and in treadmill test, preventive effect reaches
96.8%.
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-14 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-14 is bacillus subtilis Pseudomonas.
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-14 16S rDNA and withered grass
The 16S rDNA sequence homologies of bacillus are up to 99%.Comparison result is consistent with the qualification result that cell classification method obtains.
First, bio-feritlizer is produced with yeast waste liquor
Embodiment 1-1
1st, microbial inoculum：Strain fermentation culture medium prescription is soluble starch 30g, analysis for soybean powder 15g, sucrose 5g, albumen
Peptone 5g, dregs of beans 3g, K2HPO4 3g、MgSO41g, bubble enemy 0.5g, moisturizing is to 1000 milliliters.Living bacteria count in microbial inoculum
For 9.8 × 109Individual/milliliter.
The fermentation condition of bacillus subtilis LVLE14 liquid fermentations production is：PH6.5-7.5, cultivation temperature are
20-40 DEG C, originate rotating speed 100-200rpm, throughput 0.5-1vvm, cultivate 35-40 hours, fermentation middle and later periods row into gemma,
Gemma rate is up to more than 95%, and the living bacteria count in zymotic fluid is in 10,000,000,000/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 2:3；Protective agent
For sodium benzoate and molasses, ratio 1:1.
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：Fermented according to the method for embodiment 1, by biocontrol water bacillus subtilis
Bacterium solution after LVLE14 fermentations presses bacterium solution with turf：Turf=1：4 ratio is adsorbed, effective work of composite bacteria agent after mixing
Bacterium number amount is up to 1.8 × 109It is individual/gram more than, 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：Fermented according to the method for embodiment 1, by biocontrol water bacillus subtilis
Bacterium solution after LVLE14 fermentations presses bacterium solution with turf：Turf=1：5 ratio is adsorbed, effective work of composite bacteria agent after mixing
Bacterium number amount is up to 1.2 × 109It is individual/gram more than, 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, the dregs of fat are 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 dregs of fat for being cooled to normal temperature are measured, fundamental property is shown in Table 3, and content of beary metal is well below life
The professional standard of organic fertilizer, is not measured；
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 7：The bio-organic fertilizer product quality measurements of different material production
As can be seen from the above table, from the point of view of bacteriostasis rate, inside solid material, hydrolysis residue is more suitable for withered grass gemma as raw material
Bacillus.
Test example
1 material and method
1.1 test sample：LVLE14 (withered grass) zymotic fluid
1.2 test target organismses
1.2.1 bactericide tests target organismses：
Ash arrhizus bacteria-Botrytis cinerea (Bc),
Melon anthrax bacteria-Colletotrichum lagenarium (Cl),
Cucumber powdery mildew's pathogen-Sphaerotheca fuliginea (Sf),
Rhizoctonia solani Kuhn-Rhizoctonia solani (Rs),
Corn bottom class germ-Helmintosporium maydis (Hm),
Fusarium graminearum-Gibbrella zeae (Gz),
Pyricularia oryzae-Pyricuraria oryzae (Po).
1.3 test method
1.3.1 sample respectively 20 times of dilution
1.3.2 bactericide raw test method：
It is measured using the potted plant seedling method of live body.Except powdery mildew is inoculated with Powdery Mildew in one day in advance before the test during experiment
Conidial suspension, put under thermostatic chamber light, sprayed after 24h outside decoction；Remaining Bc, Cl, Rs, Hm, Gz, Po etc. are first in each basin
Plant and decoction is sprayed on crop, 4 hours or so each pathogens of inoculation, put in different temperatures or thermostatic chamber humidistat respectively after decoction is dry
Moisturizing is induced, and investigation morbidity result, is contrasted with blank control after 6 days, calculates prevention effect.
2 result of the tests
Data result is surveyed in the life of the bactericide of table 8
The bacillus subtilis LVLE14 used in the present invention is can be seen that from above test effect to be had to powdery mildew of cucumber
Obviously preventive effect, preventive effect are up to 96.8%, are also up to 74.4% to the preventive effect of gray mold of cucumber, effect highly significant.
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, have preventive effect and fertilizer efficiency concurrently.
Claims (10)
1. one plant of bacillus subtilis with suppression disease activity, it is characterised in that：The bacillus subtilis with suppression disease activity
Bacterium is bacillus subtilis (Bacillus subtilis) LVLE14, is deposited in China Committee for Culture Collection of Microorganisms
Common micro-organisms center, preservation address are Beijing；Preservation date is on June 20th, 2016, and deposit number is CGMCC
No.12639。
2. the fermentation culture method of the bacillus subtilis with suppression disease activity described in claim 1, it is characterised in that：It is described
Bacillus subtilis LVLE14 liquid fermentations production fermentation condition be：PH6.0-8.0, cultivation temperature are 20-40 DEG C, and starting turns
Fast 100-200rpm, throughput 0.5-1vvm, cultivate 35-40 hours, fermentation middle and later periods row is up to 95% into gemma, gemma rate
More than, the living bacteria count in zymotic fluid is in 10,000,000,000/more than ml.
A kind of 3. method that liquid bio fertilizer is produced with bacillus subtilis microbial agent, it is characterised in that：Will be withered described in claim 1
Careless fermentation of bacillus production, the zymotic fluid are added in liquid organic starting material by 0.1-3% mass ratio, obtain liquid life
Thing fertilizer.
4. the method according to claim 3 that liquid bio fertilizer is produced with bacillus subtilis microbial agent, it is characterised in that：Institute
State liquid organic starting material and be selected from yeast waste liquor or amino acid waste liquid.
5. the method described in claim 3 produces obtained liquid bio fertilizer, it is characterised in that：The liquid bio fertilizer is comprising withered
Careless fermentation of bacillus liquid and liquid organic starting material, the bacillus subtilis fermentation liquor account for the 0.1- of liquid bio fertilizer gross weight
3%.
A kind of 6. method that biological organic fertilizer is produced with bacillus subtilis microbial agent, it is characterised in that：By SOLID ORGANIC raw material
PH controls are in 5.0-8.0, crushing；Fermentation of bacillus subtilis described in claim 1 is produced, gained zymotic fluid adsorbing medium
Solid bacillus subtilis microbial agent is obtained after absorption, it is organic that according to 2%-10% mass ratioes solid fungicide is added into solid, powdery
In raw material, solid biologic organic fertilizer is obtained.
7. the method according to claim 6 that biological organic fertilizer is produced with bacillus subtilis microbial agent, it is characterised in that：Institute
The content of organic matter for stating SOLID ORGANIC raw material is more than 30%；The SOLID ORGANIC raw material is selected from the hydrolysis of amino acids production process
The dregs of fat of slag or edible fat production process.
8. the method described in claim 7 produces obtained biological organic fertilizer, it is characterised in that：The biological organic fertilizer includes solid
Body bacillus subtilis microbial agent and SOLID ORGANIC raw material, the solid bacillus subtilis microbial agent account for the gross weight of biological organic fertilizer
2%-10%.
9. the method that biological organic fertilizer is produced with bacillus subtilis microbial agent according to claim 3 or 6, its feature exist
In：The one or more of N,P,K fertilizers can also be added in the liquid bio fertilizer；In the biological organic fertilizer also
The one or more of N,P,K fertilizers can be added.
10. bacillus subtilis described in claim 1 especially suppresses powdery mildew of cucumber in suppression crop disease and/or suppresses yellow
Application in melon gray mold.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610756959.7A CN107779413B (en) | 2016-08-29 | 2016-08-29 | Biocontrol bacillus subtilis, production method, liquid fertilizer and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610756959.7A CN107779413B (en) | 2016-08-29 | 2016-08-29 | Biocontrol bacillus subtilis, production method, liquid fertilizer and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107779413A true CN107779413A (en) | 2018-03-09 |
| CN107779413B CN107779413B (en) | 2021-01-29 |
Family
ID=61449749
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610756959.7A Active CN107779413B (en) | 2016-08-29 | 2016-08-29 | Biocontrol bacillus subtilis, production method, liquid fertilizer and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107779413B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN108841744A (en) * | 2018-06-15 | 2018-11-20 | 保定微控生物科技有限公司 | A kind of bacillus subtilis with diseases prevention and degradation Phos double action |
| CN108977393A (en) * | 2018-08-14 | 2018-12-11 | 上海绿乐生物科技有限公司 | A method of microbial bacterial agent and biological organic fertilizer are produced using edible oil organic carbon filter cake |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1142988A1 (en) * | 1999-01-14 | 2001-10-10 | Gold Kosan Co. Ltd | Novel microorganism and use thereof |
| CN102442859A (en) * | 2010-10-12 | 2012-05-09 | 上海绿乐生物科技有限公司 | Method for producing compound microbial fertilizer with microbial inoculum permeation and adsorption method |
| CN103952328A (en) * | 2013-11-13 | 2014-07-30 | 陕西省微生物研究所 | Bacillus subtilis strain A33, microbial inoculum and preparation method and application thereof |
| CN104973960A (en) * | 2015-06-26 | 2015-10-14 | 上海诺同农业科技有限公司 | Fruit tree dedicated bio-organic fertilizer, production method and application method thereof |
-
2016
- 2016-08-29 CN CN201610756959.7A patent/CN107779413B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1142988A1 (en) * | 1999-01-14 | 2001-10-10 | Gold Kosan Co. Ltd | Novel microorganism and use thereof |
| CN102442859A (en) * | 2010-10-12 | 2012-05-09 | 上海绿乐生物科技有限公司 | Method for producing compound microbial fertilizer with microbial inoculum permeation and adsorption method |
| CN103952328A (en) * | 2013-11-13 | 2014-07-30 | 陕西省微生物研究所 | Bacillus subtilis strain A33, microbial inoculum and preparation method and application thereof |
| CN104973960A (en) * | 2015-06-26 | 2015-10-14 | 上海诺同农业科技有限公司 | Fruit tree dedicated bio-organic fertilizer, production method and application method thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN108841744A (en) * | 2018-06-15 | 2018-11-20 | 保定微控生物科技有限公司 | A kind of bacillus subtilis with diseases prevention and degradation Phos double action |
| CN108977393A (en) * | 2018-08-14 | 2018-12-11 | 上海绿乐生物科技有限公司 | A method of microbial bacterial agent and biological organic fertilizer are produced using edible oil organic carbon filter cake |
| CN108977393B (en) * | 2018-08-14 | 2021-11-26 | 上海绿乐生物科技有限公司 | Method for producing microbial agent and biological organic fertilizer by using edible oil organic carbon filter cake |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107779413B (en) | 2021-01-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104672009B (en) | A kind of multifunction activity organic fertilizer and preparation method thereof | |
| CN100334201C (en) | Bacillus subtilis and its uses | |
| CN105018366B (en) | Methylotrophic bacillus and its application | |
| CN106591185A (en) | Application and preparation of bacillus amyloliquefaciens subsp. plantarum and bacterial agent thereof | |
| CN101875571B (en) | Method for preparing enhanced liquid microbial organic fertilizer | |
| CN104211504A (en) | Compound microorganism environment-friendly fertilizer and preparation method thereof | |
| CN104894010A (en) | Compound microbial fertilizer for antagonism of soil-borne fungal diseases, and preparation method and application thereof | |
| CN104498386A (en) | Preparation method and applications of wild jujube endophytic bacillus amyloliquefaciens new strain SZ23 and fermentation broth | |
| CN106591193A (en) | Bacillus amyloliquefaciens with broad spectrum growth-promoting and stress-resisting effects | |
| CN101709279B (en) | Pseudomonas putida and spray agent thereof | |
| CN1861550A (en) | Ginseng genus micro ecological active fertilizer | |
| US10807920B2 (en) | Trichoderma-derived selenium nanoparticles foliar fertilizer for reducing crop fungal diseases and toxin contamination | |
| CN110305671A (en) | One kind is for facilities vegetable continuous cropping soil biological modification agent and its preparation and application method | |
| CN107325821A (en) | A kind of biomass carbon base microbe microbial inoculum for preventing and treating watermelon blight and preparation method and application | |
| CN113862189A (en) | Corn growth promoting, probiotic and biocontrol bacterial fertilizer integrated preparation and preparation method thereof | |
| CN101463339B (en) | Inocula for preventing apple fungous diseases and increasing yield, and special strain therefore and use thereof | |
| CN106146194B (en) | A kind of anti-continuous cropping microbial inoculum | |
| CN106011030B (en) | A kind of mutagenic strain Bs6602v and its application | |
| CN101974454B (en) | Degrading bacteria for pyrethroids insecticides and bactericide thereof | |
| CN101974462B (en) | Bacillus subtilis for preventing and controlling Chinese herbal medicine soil-borne diseases and preparation method of bactericide thereof | |
| CN107779413A (en) | One plant of biocontrol water bacillus subtilis, production method, liquid fertilizer and application | |
| CN102586130B (en) | Bio-control bacterial strain and bio-control bactericide capable of preventing grape anthracnose and application thereof | |
| KR100851399B1 (en) | A composition for biological control of pepper blight caused by Capsicum annuum L. using Trichoderma harzianum having antagonistic activity | |
| CN109734489A (en) | A kind of ferment probiotics multi-functional bio-fertilizer | |
| CN109957517B (en) | A kind of A Si Penicillium notatum and its microbial inoculum and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |