CN106348886A - Preparation method of bacillus megaterium inoculum-containing compound fertilizer - Google Patents
Preparation method of bacillus megaterium inoculum-containing compound fertilizer Download PDFInfo
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- CN106348886A CN106348886A CN201610776246.7A CN201610776246A CN106348886A CN 106348886 A CN106348886 A CN 106348886A CN 201610776246 A CN201610776246 A CN 201610776246A CN 106348886 A CN106348886 A CN 106348886A
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- 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
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
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- 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
- C05G5/30—Layered or coated, e.g. dust-preventing coatings
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention relates to a preparation method of a bacillus megaterium inoculum-containing compound fertilizer. The preparation method comprises the following steps: grinding dried algae into powder, adding the alga powder into water according to a certain ratio, putting into a fermenting tank, and sterilizing for a certain time at a certain temperature; preparing seed liquid: screening to obtain a fermenting strain YIC-BM1 bacillus megaterium capable of producing cellulose, inoculating a liquid culture medium with the fermenting strain YIC-BM1, and culturing for a certain time; adding the seed liquid into the fermenting tank according to a certain mass ratio, fermenting under a certain condition, adjusting a fermenting condition, keeping for a certain time, adjusting the fermenting condition again and supplementing the seed liquid, adjusting for three times, airing the fermented liquid to obtain a bacillus megaterium inoculum, and adding the bacillus megaterium inoculum capable of producing the cellulose into a compound fertilizer in an externally enwrapping way to obtain the cellulose-producing bacillus megaterium inoculum-containing alga compound fertilizer. By the production method, the fermenting process is stable and controllable, the fermenting cycle is short, and the product efficacy is remarkable; the production method is suitable for large-scale production.
Description
Technical field
The present invention relates to a kind of preparation method of the Chemical Mixed Fertilizer containing microbial bacterial agent.
Background technology
Marine alga fertilizer, with natural seaweed as raw material, is processed through special biochemical process, extracts and retain marine alga natural activity
Composition, a kind of novel organic fertilizer material that refine concentrates and formed.Marine alga fertilizer has raising crop yield and quality, resists disease pest
Effect that evil and poor environment and improved soil conserve water and soil, it is applied to agricultural production the huge market demand and good
Development prospect.But, the quality of marine alga fertilizer to be ensured, it is necessary for getting hold of Sargassum raw material and the crucial ring of extraction process two
Section.
First, the quality of marine alga fertilizer quality depends on raw material used.At present, the raw material used by alga fertilizer manufacturing enterprise
Mainly based on Brown algae, including yellow tang, Thallus Laminariae (Thallus Eckloniae), Thallus Laminariae (Thallus Eckloniae), Alga Sgrgassi Enervess etc..Other algae such as Fucus Vesiculosus, Entermorpha, ulva, Kappa
Zao Dengyeyou minority enterprise with.External alga fertilizer enterprise is most to be raw material using Brown algae, is wherein especially applied with yellow tang
The most extensive such as famous brand name such as seasol, maxicrop, practical function effect also indicates that yellow tang is the Sargassum raw material of high-quality.
But, China coast does not have yellow tang to be distributed, and the raw material type that domestic enterprise adopts is numerous and diverse, and quality is irregular to differ, and causes Sargassum
Fertilizer product quality is very different, or even some producers are turned on the camouflage of " alga fertilizer ", directly blends alginic acid, fulvic acid, nitrogen phosphorus
Potassium etc. pretends to be alga fertilizer, have impact on alga fertilizer quality, has upset alga fertilizer market.
Secondly, the second factor of decision alga fertilizer quality is extraction process.Different extraction processes is to alga fertilizer activity
The species and content impact of composition is huge.At present, the extracting method of Sargassum active component mainly has chemical method, Physical and life
Thing method.Chemical method mainly uses acid, alkali and organic solvent and processes alginic cell, so that cell is cleared up or endogenous substance solubilising, should
Method is simple to operate, easily realizes, and is also the method that most alga fertilizer manufacturing enterprises adopt both at home and abroad.But, chemistry examination
Agent is sizable to the destruction of the active material ingredients in alginic cell, and the organic reagent remaining is also potential to environment
Harm.The principle of Physical is control pressure, the physical environmental condition such as temperature, so that alginic cell is crushed using mechanical force, interior
Tolerant release.Although the method technique cleaning, environmental friendliness, the requirement for instrument and equipment is harsh, change in reaction conditions
Acutely, relatively costly it is difficult to realize large-scale production.Bioanalysises include two methods of enzymic degradation and fermentable, its master
Principle is wanted to be to destroy the small-molecule substance that alginic cell wall construction macromolecular substances of degrading easily absorb, the party for plant
Method reaction is gentle, product diversification, and whole production process safety and environmental protection is pollution-free, is preferable Sargassum fertilizer production method.
Research using Production by Microorganism Fermentation marine alga fertilizer is relatively fewer, and only several researchs are from multiple micro-
Biology carries out similar compost fermentation to Sargassum raw material or aerobic fermentation is processed, and there is sweat poor controllability, fermentation period is long,
The problems such as target product is indefinite is it is difficult to meet the unification of industrialization production requirements, stable, controlled standard, the quality of product
It is difficult to ensure.
Evaluation index for agricultural micro organism mainly includes the microbial inoculum quantity in the purity and unit mass of microbial inoculum.Cause
This, the bacterial preparation process as prior art is usually: carries out once under the conditions of the temperature of the suitable existence of microorganism and ph
Fermentation technology, to reach the maximization of bacterium quantity in unit mass, is equivalent to hot-house culture.
Because the acidifying of current soil compaction is serious, outside air temperature is changeable, and the microbial bacterial agent of hot-house culture is directly thrown
It is put in soil, the soil environment because not adapting to severe causes mortality, even if having surviving because population cannot be formed on a small quantity
Effect, also cannot play the effect of microbial bacterial agent.
At present no the bacillus megaterium of cellulase-producing and Chemical Mixed Fertilizer most economical under conditions of play maximum fertilizer efficiency
Good mass ratio.
Content of the invention
The technical problem to be solved is to provide a kind of preparation of the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum
Method, prepares Chemical Mixed Fertilizer using bacillus megaterium, and the method makes bacterium have preferable environment adaptation by adjusting fermentation condition
Ability is it is easy to play the advantage of bacterium in severe soil environment；By determining the optimal matter of bacillus megaterium and Chemical Mixed Fertilizer
Maximum fertilizer efficiency is played under conditions of amount is more most economical than reaching.
In order to solve above-mentioned technical problem, the present invention employs the following technical solutions:
A kind of preparation method of the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum is it is characterised in that follow the steps below:
Nitrogenous fertilizer, phosphate fertilizer, potash fertilizer and adjuvant are added to pelletizing drum according to mass ratio 10～90:10～90:10～90:50～70
Middle pelletize, then after drying, cooling and screening, obtains complex fertilizer；In pack roller, oily and huge spore bar will be wrapped up
Bacteria agent is injected to Chemical Mixed Fertilizer surface, obtains the bacillus megaterium Chemical Mixed Fertilizer containing cellulase-producing, wherein Chemical Mixed Fertilizer, parcel
The mass ratio of the bacillus megaterium microbial inoculum of oil and cellulase-producing is 55～99.8:0.1～5:0.1～40；
Described nitrogenous fertilizer is the two or more of carbamide, ammonium chloride, ammonium nitrate p and ses acid one of ammonium or arbitrary proportion；
Described phosphate fertilizer is the two or more of one of MAP, Diammonium phosphate (DAP) and potassium dihydrogen phosphate or arbitrary proportion；
Described potash fertilizer is the two or more of one of potassium sulfate, potassium nitrate and potassium chloride or arbitrary proportion；
Described adjuvant is the two or more of one of bentonite, Kaolin and Pulvis Talci or arbitrary proportion；
Wherein bacillus megaterium microbial inoculum is prepared from according to following steps:
1), prepare marine algae powder: the Sargassum after drying is clayed into power；
2), prepare product a: marine algae powder and water and load in fermentation tank according to mass ratio 1～10:100, in 100～150 DEG C of conditions
Lower sterilizing 15～35min, obtains product a；
And prepare seed liquor: bacillus megaterium is seeded to fluid medium, cultivates 20～30 hours at 25 DEG C～40 DEG C；Huge
Volume ratio between Bacterium anthracoides and culture medium is 1～10:100；
3), prepare product b: seed liquor is added in product a and obtains product b, wherein seed liquor and the mass ratio of product a are 1
～10:100；
4), prepare product c: be 25 DEG C～40 DEG C in temperature, under conditions of ph5～9, product b fermented, fermentation time is 6
～10 days, obtain product c；
5), prepare product d: the ph to 5.0～6.0 of adjustment product c, temperature, to 3～10 DEG C, preserves 3h～72h, obtains product d；
6), prepare product e: the temperature of adjustment product d to 25 DEG C～40 DEG C, ph to 5～9, then supplements seed liquor, the kind supplemented
Sub- liquid is 1～10:100 with the mass ratio of product d, preserves 3h～72h, obtains product e；
7), prepare product f: the ph to 5.0～6.0 of adjustment product e, temperature, to 3～10 DEG C, preserves 3h～72h, obtains product f；
8), prepare product g: the temperature of adjustment product f to 25 DEG C～40 DEG C, ph to 5～9, then supplements seed liquor, the kind supplemented
Sub- liquid is 1～10:100 with the mass ratio of product f, preserves 3h～72h, obtains product g；
9), prepare bacillus megaterium microbial inoculum: after product g is air-dried, obtain bacillus megaterium microbial inoculum.
Described fluid medium is made into according to following mass ratio: 5 parts of sodium alginate, 5 parts of ammonium sulfate, 1 part of magnesium sulfate, phosphoric acid
2 parts of hydrogen dipotassium, 0.01 part of ferrous sulfate, 20 parts of agar and 1000 parts of distilled water.
Step 2) mass ratio of described marine algae powder and water is 4:100；115 DEG C of sterilising temp, sterilization time 25min；System
During standby seed liquor, bacillus megaterium is seeded in fluid medium, cultivates 24 hours at 30 DEG C, obtains seed liquor, huge bud
The volume ratio of spore bacillus and culture medium is 5:100；In step 3), seed liquor is 10:100 with the mass ratio of product a；Step 5)
In step 7), temperature is 5 DEG C, and the holding time is 12h.
In step 4), step 6) and step 8), temperature is 30 DEG C, and ph is 6.5～7.0；Step 4) fermentation time is 8 days；
Step 6) and step 8) holding time are 12h；Mend in the seed liquor supplemented in step 6) and the mass ratio of product d and step 8)
The seed liquor filled is 5:100 with the mass ratio of product f.
In step 5), step 6) step 7) and step 8), adjustment temperature is and adjusts 3 DEG C per hour.
The mass ratio of the bacillus megaterium microbial inoculum of described Chemical Mixed Fertilizer, parcel oil and cellulase-producing is 99.5:0.2:
0.3.
Described Sargassum refers to the two or more of sargassum thunbergii, one of Sargassum phyllocystum Tseng et Lu,Sargassum horneri (Turn.) C. Ag. (Fucus horneri (Turn.)C.Ag.,Spongocarpus horneri Kutz.) and Alga Sgrgassi Enervess or arbitrary proportion.
Described bacillus megaterium is bacillus megaterium yic～bm1, and preserving number is cgmcc no.12156.
The present invention has following Advantageous Effects:
1st, adopt the bacillus megaterium of the present invention to fermentation of seaweed, sweat is stablized controlled, and fermentation period is short, product efficiency
Significantly, it is suitable to large-scale production.
2nd, the fermentation liquid that the present invention obtains has good water solubility it is easy to plant absorption utilizes, simultaneously as a kind of important
Signaling molecule, is capable of the growth regulating of involved in plant and induction resistance mechanism promotes the growth of plant, improves plant to bad ring
Border and the resistance of pest and disease damage.
3rd, the present invention makes the bacterium that fermentation goes out possess the ability adapting to severe soil environment by adjusting fermentation condition, favorably
Performance in bacterium function.
4th, the present invention, by slow temperature adjustment, makes the change of the bacillus megaterium adaptive temperature of cellulase-producing,
Avoid temperature change too fast, the bacillus megaterium of cellulase-producing does not adapt to and mortality.
5th, the present invention passes through parcel oil by the bacillus megaterium of cellulase-producing and Chemical Mixed Fertilizer obstruct, and using parcel
The bacillus megaterium of cellulase-producing is uniformly bonded in the surface of fertilizer by the viscosity of oil, can be prevented effectively from product cellulose
The bacillus megaterium of enzyme and highly-saline Chemical Mixed Fertilizer directly contact, prevent from leading to cellulase-producing because Chemical Mixed Fertilizer salinity is too high
Bacillus megaterium cell dehydration, causes the bacillus megaterium mortality of cellulase-producing.
6th, the present invention determines the bacillus megaterium of cellulase-producing and the best in quality of Chemical Mixed Fertilizer by many experiments
Than.
Brief description
Fig. 1 is the Gram’s staining of microexamination yic～bm1 bacterial strain and assembles agglomerating phenomenon figure.
Yic～bm1 bacterial strain phylogenetic tree schematic diagram based on 16srdna sequence construct for the Fig. 2.
Fig. 3 is the schematic diagram of the impact to yic～bm1 strain enzyme-producing for the different temperatures.
Fig. 4 is the impact schematic diagram to yic～bm1 strain enzyme-producing for the different ph.
Specific embodiment
Further illustrate the present invention with reference to instantiation.
The following is Sargassum liquid preparation embodiment of the present invention.
Embodiment 1
Fresh sargassum thunbergii after clean drying is worn into 50～60 mesh powder；
Marine algae powder loads in fermentation tank with water according to mass ratio 4:100, and sterilize under the conditions of 115 DEG C 25min, obtains product a；
And prepare seed liquor: the bacillus megaterium yic～bm1 of cellulase-producing is seeded to fluid medium, cultivates 24 at 35 DEG C
Hour；The preserving number of the bacillus megaterium yic～bm1 of described cellulase-producing is cgmcc no.12156；Cellulase-producing
Bacillus megaterium yic～bm1 and culture medium volume ratio be 5:100.Seed liquor is added in product a, wherein plants
Sub- liquid is 10:100 with the mass ratio of product a, obtains product b；It is 35 DEG C in temperature, to product b under conditions of ph6.0～6.5
Fermented, fermentation time is 8 days, obtains product c；The ph to 5.5～6.0 of adjustment product c, and with per hour 3 DEG C of temperature
Change adjustment temperature, to 5 DEG C, preserves 12h, obtains product d；With the temperature of per hour 3 DEG C of temperature change adjustment product d to 35
DEG C, ph to 6.0～6.5, then supplements seed liquor, the seed liquor supplemented is 6:100 with the mass ratio of product d, preserves 12h, obtains
To product e；The ph to 5.5～6.0 of adjustment product e, and with per hour 3 DEG C of temperature change adjustment temperature to 5 DEG C, preserve 12h,
Obtain product f；The temperature of product f is adjusted to 35 DEG C with per hour 3 DEG C of temperature change, ph to 6.0～6.5, then supplements and plants
Sub- liquid, the seed liquor supplemented is 6:100 with the mass ratio of product f, preserves 12h, obtains product g；Produced after product g is air-dried
The bacillus megaterium microbial inoculum of cellulase；Nitrogenous fertilizer, phosphate fertilizer, potash fertilizer and adjuvant are added to according to mass ratio 20:20:10:50
Pelletize in pelletizing drum, after product drying, cooling and screening, enters into pack roller, obtains complex fertilizer；In pack roller
In, the bacillus megaterium microbial inoculum of parcel oil and cellulase-producing is injected to Chemical Mixed Fertilizer surface, complex fertilizer, parcel are oily and produce
The mass ratio of the bacillus megaterium microbial inoculum of cellulase is 99.5:0.2:0.3, obtains the huge spore containing cellulase-producing
The seaweed compound fertilizer of bacillus.
Embodiment 2
Fresh sargassum thunbergii after clean drying is worn into 50～60 mesh powder；
Marine algae powder loads in fermentation tank with water according to mass ratio 4:100, and sterilize under the conditions of 115 DEG C 25min, obtains product a；
And prepare seed liquor: the bacillus megaterium yic～bm1 of cellulase-producing is seeded to fluid medium, cultivates 24 at 35 DEG C
Hour；The preserving number of the bacillus megaterium yic～bm1 of described cellulase-producing is cgmcc no.12156；Cellulase-producing
Bacillus megaterium yic～bm1 and culture medium volume ratio be 5:100.Seed liquor is added in product a, wherein plants
Sub- liquid is 10:100 with the mass ratio of product a, obtains product b；It is 35 DEG C in temperature, to product b under conditions of ph6.0～6.5
Fermented, fermentation time is 8 days, obtains product c；The ph to 5.5～6.0 of adjustment product c, and with per hour 3 DEG C of temperature
Change adjustment temperature, to 5 DEG C, preserves 12h, obtains product d；With the temperature of per hour 3 DEG C of temperature change adjustment product d to 35
DEG C, ph to 6.0～6.5, then supplements seed liquor, the seed liquor supplemented is 6:100 with the mass ratio of product d, preserves 12h, obtains
To product e；The ph to 5.5～6.0 of adjustment product e, and with per hour 3 DEG C of temperature change adjustment temperature to 5 DEG C, preserve 12h,
Obtain product f；The temperature of product f is adjusted to 35 DEG C with per hour 3 DEG C of temperature change, ph to 6.0～6.5, then supplements and plants
Sub- liquid, the seed liquor supplemented is 6:100 with the mass ratio of product f, preserves 12h, obtains product g；Produced after product g is air-dried
The bacillus megaterium microbial inoculum of cellulase；Nitrogenous fertilizer, phosphate fertilizer, potash fertilizer and adjuvant are added to according to mass ratio 20:20:10:50
Pelletize in pelletizing drum, after product drying, cooling and screening, enters into pack roller, obtains complex fertilizer；In pack roller
In, the bacillus megaterium microbial inoculum of parcel oil and cellulase-producing is injected to Chemical Mixed Fertilizer surface, complex fertilizer, parcel are oily and produce
The mass ratio of the bacillus megaterium microbial inoculum of cellulase is 99.7:0.2:0.1, obtains the huge spore containing cellulase-producing
The seaweed compound fertilizer of bacillus.
Embodiment 3
Fresh sargassum thunbergii after clean drying is worn into 50～60 mesh powder；
Marine algae powder loads in fermentation tank with water according to mass ratio 4:100, and sterilize under the conditions of 115 DEG C 25min, obtains product a；
And prepare seed liquor: the bacillus megaterium yic～bm1 of cellulase-producing is seeded to fluid medium, cultivates 24 at 35 DEG C
Hour；The preserving number of the bacillus megaterium yic～bm1 of described cellulase-producing is cgmcc no.12156；Cellulase-producing
Bacillus megaterium yic～bm1 and culture medium volume ratio be 5:100.Seed liquor is added in product a, wherein plants
Sub- liquid is 10:100 with the mass ratio of product a, obtains product b；It is 35 DEG C in temperature, to product b under conditions of ph6.0～6.5
Fermented, fermentation time is 8 days, obtains product c；The ph to 5.5～6.0 of adjustment product c, and with per hour 3 DEG C of temperature
Change adjustment temperature, to 5 DEG C, preserves 12h, obtains product d；With the temperature of per hour 3 DEG C of temperature change adjustment product d to 35
DEG C, ph to 6.0～6.5, then supplements seed liquor, the seed liquor supplemented is 6:100 with the mass ratio of product d, preserves 12h, obtains
To product e；The ph to 5.5～6.0 of adjustment product e, and with per hour 3 DEG C of temperature change adjustment temperature to 5 DEG C, preserve 12h,
Obtain product f；The temperature of product f is adjusted to 35 DEG C with per hour 3 DEG C of temperature change, ph to 6.0～6.5, then supplements and plants
Sub- liquid, the seed liquor supplemented is 6:100 with the mass ratio of product f, preserves 12h, obtains product g；Produced after product g is air-dried
The bacillus megaterium microbial inoculum of cellulase；Nitrogenous fertilizer, phosphate fertilizer, potash fertilizer and adjuvant are added to according to mass ratio 20:20:10:50
Pelletize in pelletizing drum, after product drying, cooling and screening, enters into pack roller, obtains complex fertilizer；In pack roller
In, the bacillus megaterium microbial inoculum of parcel oil and cellulase-producing is injected to Chemical Mixed Fertilizer surface, complex fertilizer, parcel are oily and produce
The mass ratio of the bacillus megaterium microbial inoculum of cellulase is 99.6:0.2:0.2, obtains the huge spore containing cellulase-producing
The seaweed compound fertilizer of bacillus.
Embodiment 4
Fresh sargassum thunbergii after clean drying is worn into 50～60 mesh powder；
Marine algae powder loads in fermentation tank with water according to mass ratio 4:100, and sterilize under the conditions of 115 DEG C 25min, obtains product a；
And prepare seed liquor: the bacillus megaterium yic～bm1 of cellulase-producing is seeded to fluid medium, cultivates 24 at 35 DEG C
Hour；The preserving number of the bacillus megaterium yic～bm1 of described cellulase-producing is cgmcc no.12156；Cellulase-producing
Bacillus megaterium yic～bm1 and culture medium volume ratio be 5:100.Seed liquor is added in product a, wherein plants
Sub- liquid is 10:100 with the mass ratio of product a, obtains product b；It is 35 DEG C in temperature, to product b under conditions of ph6.0～6.5
Fermented, fermentation time is 8 days, obtains product c；The ph to 5.5～6.0 of adjustment product c, and with per hour 3 DEG C of temperature
Change adjustment temperature, to 5 DEG C, preserves 12h, obtains product d；With the temperature of per hour 3 DEG C of temperature change adjustment product d to 35
DEG C, ph to 6.0～6.5, then supplements seed liquor, the seed liquor supplemented is 6:100 with the mass ratio of product d, preserves 12h, obtains
To product e；The ph to 5.5～6.0 of adjustment product e, and with per hour 3 DEG C of temperature change adjustment temperature to 5 DEG C, preserve 12h,
Obtain product f；The temperature of product f is adjusted to 35 DEG C with per hour 3 DEG C of temperature change, ph to 6.0～6.5, then supplements and plants
Sub- liquid, the seed liquor supplemented is 6:100 with the mass ratio of product f, preserves 12h, obtains product g；Produced after product g is air-dried
The bacillus megaterium microbial inoculum of cellulase；Nitrogenous fertilizer, phosphate fertilizer, potash fertilizer and adjuvant are added to according to mass ratio 20:20:10:50
Pelletize in pelletizing drum, after product drying, cooling and screening, enters into pack roller, obtains complex fertilizer；In pack roller
In, the bacillus megaterium microbial inoculum of parcel oil and cellulase-producing is injected to Chemical Mixed Fertilizer surface, complex fertilizer, parcel are oily and produce
The mass ratio of the bacillus megaterium microbial inoculum of cellulase is 99.4:0.2:0.4, obtains the huge spore containing cellulase-producing
The seaweed compound fertilizer of bacillus.
Embodiment 5
Fresh sargassum thunbergii after clean drying is worn into 50～60 mesh powder；
Marine algae powder loads in fermentation tank with water according to mass ratio 4:100, and sterilize under the conditions of 115 DEG C 25min, obtains product a；
And prepare seed liquor: the bacillus megaterium yic～bm1 of cellulase-producing is seeded to fluid medium, cultivates 24 at 35 DEG C
Hour；The preserving number of the bacillus megaterium yic～bm1 of described cellulase-producing is cgmcc no.12156；Cellulase-producing
Bacillus megaterium yic～bm1 and culture medium volume ratio be 5:100.Seed liquor is added in product a, wherein plants
Sub- liquid is 10:100 with the mass ratio of product a, obtains product b；It is 35 DEG C in temperature, to product b under conditions of ph6.0～6.5
Fermented, fermentation time is 8 days, obtains product c；The ph to 5.5～6.0 of adjustment product c, and with per hour 3 DEG C of temperature
Change adjustment temperature, to 5 DEG C, preserves 12h, obtains product d；With the temperature of per hour 3 DEG C of temperature change adjustment product d to 35
DEG C, ph to 6.0～6.5, then supplements seed liquor, the seed liquor supplemented is 6:100 with the mass ratio of product d, preserves 12h, obtains
To product e；The ph to 5.5～6.0 of adjustment product e, and with per hour 3 DEG C of temperature change adjustment temperature to 5 DEG C, preserve 12h,
Obtain product f；The temperature of product f is adjusted to 35 DEG C with per hour 3 DEG C of temperature change, ph to 6.0～6.5, then supplements and plants
Sub- liquid, the seed liquor supplemented is 6:100 with the mass ratio of product f, preserves 12h, obtains product g；Produced after product g is air-dried
The bacillus megaterium microbial inoculum of cellulase；Nitrogenous fertilizer, phosphate fertilizer, potash fertilizer and adjuvant are added to according to mass ratio 20:20:10:50
Pelletize in pelletizing drum, after product drying, cooling and screening, enters into pack roller, obtains complex fertilizer；In pack roller
In, the bacillus megaterium microbial inoculum of parcel oil and cellulase-producing is injected to Chemical Mixed Fertilizer surface, complex fertilizer, parcel are oily and produce
The mass ratio of the bacillus megaterium microbial inoculum of cellulase is 99.3:0.2:0.5, obtains the huge spore containing cellulase-producing
The seaweed compound fertilizer of bacillus.
In embodiment 1 to 5, described nitrogenous fertilizer is ammonium sulfate, and phosphate fertilizer is MAP, and potash fertilizer is potassium sulfate, and adjuvant is swelling
Soil.
1 materials and methods:
1.1 test sites: Wuzhoufeng Agricultural Technology Co., Ltd. proving ground, neutral brown earth, middle high fertility.
1.2 tomato varieties: tobacco powder 2.
1.3 for examination fertilizer: the seaweed compound fertilizer of the bacillus megaterium containing cellulase-producing, by the rich agricultural science and technology in the Five continents
Company limited provides；Common Chemical Mixed Fertilizer, is provided by Wuzhoufeng Agricultural Technology Co., Ltd..
1.4 EXPERIMENTAL DESIGN: test and set 6 process:
Process 1: (embodiment nitrogen, phosphorus, potassium content be consistent and common compound that be added without bacillus megaterium microbial inoculum for comparative example
Fertile)；
Process 2: the seaweed compound fertilizer of the bacillus megaterium containing cellulase-producing described in embodiment 1；
Process 3: the seaweed compound fertilizer of the bacillus megaterium containing cellulase-producing described in embodiment 2；
Process 4: the seaweed compound fertilizer of the bacillus megaterium containing cellulase-producing described in embodiment 3；
Process 5: the seaweed compound fertilizer of the bacillus megaterium containing cellulase-producing described in embodiment 4；
Process 6: the seaweed compound fertilizer of the bacillus megaterium containing cellulase-producing described in embodiment 5.
Each processes 3 repetitions, random district's groups arrangement, plot area 15m2, it is colonized 75 plants, if protection row.Cell list is received
Calculate yield respectively.
1.5 fertilizing methods:
Each process 1 mu and apply 100kg Chemical Mixed Fertilizer make base manure, other management are with conventional.
2 results and analysis
2.1 Fructus Lycopersici esculenti different fertilization fertilizer effect analyses
The impact to tomato yield for table 1 different disposal
As can be seen from Table 1, each embodiment is compared with comparative example, all has different degrees of volume increase.And embodiment 1 effect of increasing production
Significantly better than embodiment 2 and embodiment 3, although and embodiment 4 and embodiment 5 effect are slightly better than embodiment 1, effect of increasing production carries
High inconspicuous.Embodiment 2, embodiment 3 and embodiment 1 be can be seen that by embodiment 1 to 5 effect huge with cellulase-producing
The increase fertilizer efficiency of Bacterium anthracoides ratio increases, and embodiment 1, embodiment 4 and embodiment 5 although cellulase-producing huge
Bacillus cereuss adding proportion is in increase, but no obvious effect of increasing production in fertilizer efficiency, economically consider compound in embodiment 1
The ratio most preferably 0.3% of the fertile bacillus megaterium with cellulase-producing.
The preserving number of described bacillus megaterium yic～bm1 for cgmcc no.12156 depositary institution title is: China is micro-
Biological inoculum preservation administration committee common micro-organisms center；Depositary institution address is: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City
No. 3；Preservation date: on March 2nd, 2016；Latin Classification And Nomenclature is:bacillus megaterium.
The biological nature of described bacillus megaterium yic～bm1 is as follows:
Cellulose culture medium: Carboxymethyl cellulose sodium 15g, ammonium nitrate 1g, magnesium sulfate 1g, potassium dihydrogen phosphate 2g, agar 20g, steam
Distilled water 1000ml.
Liquid fermentation medium: Carboxymethyl cellulose sodium 10g, peptone 3g, yeast powder 0.5g, ammonium nitrate 2g, magnesium sulfate
0.3g, potassium dihydrogen phosphate 2g, calcium chloride 0.3g, distilled water 1000ml, ph7.0-7.4.
After yic～bm1 bacterial strain grows 48h on cellulose culture medium flat plate, bacterium colony is circular, white, surfacing.Micro-
Sem observation bacterial strain is Gram-positive, shaft-like, can move, and produces spore.20～40 DEG C of growth temperature range, ph scope 6-10,
Nacl concentration range 0～20%, 37 DEG C of bacterial strain optimum growth temperature, the most suitable growth ph 7.5.Catalase, oxidase, tilactase
The positive, indole, vp experiment is negative, does not go back orthonitric acid；Glucose, mannose, Lactose, Raffinose, sucrose, Arab can be utilized
Sugar, xylose, Mannitol, Sorbitol.According to its physiological and biochemical property, this identification of strains is bacillus megateriumbacillus megaterium.
Fig. 1 finds out, yic～bm1 bacterial strain is gram positive bacteria；
Fig. 2 finds out, the sibship of yic～bm1 bacterial strain is nearest with bacillus megaterium；
Fig. 3 finds out, the most suitable producing enzyme temperature of yic～bm1 bacterial strain is 35 DEG C；
Fig. 4 finds out, optimal producing enzyme ph of yic～bm1 bacterial strain is 6.0.
Claims (9)
1. a kind of preparation method of the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum is it is characterised in that follow the steps below:
Nitrogenous fertilizer, phosphate fertilizer, potash fertilizer and adjuvant are added to pelletizing drum according to mass ratio 10～90:10～90:10～90:50～70
Middle pelletize, then after drying, cooling and screening, obtains complex fertilizer；In pack roller, oily and huge spore bar will be wrapped up
Bacteria agent is injected to Chemical Mixed Fertilizer surface, obtains the bacillus megaterium Chemical Mixed Fertilizer containing cellulase-producing, wherein Chemical Mixed Fertilizer, parcel
The mass ratio of the bacillus megaterium microbial inoculum of oil and cellulase-producing is 55～99.8:0.1～5:0.1～40；
Described nitrogenous fertilizer is the two or more of carbamide, ammonium chloride, ammonium nitrate p and ses acid one of ammonium or arbitrary proportion；
Described phosphate fertilizer is the two or more of one of MAP, Diammonium phosphate (DAP) and potassium dihydrogen phosphate or arbitrary proportion；
Described potash fertilizer is the two or more of one of potassium sulfate, potassium nitrate and potassium chloride or arbitrary proportion；
Described adjuvant is the two or more of one of bentonite, Kaolin and Pulvis Talci or arbitrary proportion；
Wherein bacillus megaterium microbial inoculum is prepared from according to following steps:
1), prepare marine algae powder: the Sargassum after drying is clayed into power；
2), prepare product a: marine algae powder and water and load in fermentation tank according to mass ratio 1～10:100, in 100～150 DEG C of conditions
Lower sterilizing 15～35min, obtains product a；
And prepare seed liquor: bacillus megaterium is seeded to fluid medium, cultivates 20～30 hours at 25 DEG C～40 DEG C；Huge
Volume ratio between Bacterium anthracoides and culture medium is 1～10:100；
3), prepare product b: seed liquor is added in product a and obtains product b, wherein seed liquor and the mass ratio of product a are 1
～10:100；
4), prepare product c: be 25 DEG C～40 DEG C in temperature, under conditions of ph5～9, product b fermented, fermentation time is 6
～10 days, obtain product c；
5), prepare product d: the ph to 5.0～6.0 of adjustment product c, temperature, to 3～10 DEG C, preserves 3h～72h, obtains product d；
6), prepare product e: the temperature of adjustment product d to 25 DEG C～40 DEG C, ph to 5～9, then supplements seed liquor, the kind supplemented
Sub- liquid is 1～10:100 with the mass ratio of product d, preserves 3h～72h, obtains product e；
7), prepare product f: the ph to 5.0～6.0 of adjustment product e, temperature, to 3～10 DEG C, preserves 3h～72h, obtains product f；
8), prepare product g: the temperature of adjustment product f to 25 DEG C～40 DEG C, ph to 5～9, then supplements seed liquor, the kind supplemented
Sub- liquid is 1～10:100 with the mass ratio of product f, preserves 3h～72h, obtains product g；
9), prepare bacillus megaterium microbial inoculum: after product g is air-dried, obtain bacillus megaterium microbial inoculum.
2. the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum as claimed in claim 1 preparation method it is characterised in that: described
Fluid medium is made into according to following mass ratio: 5 parts of sodium alginate, 5 parts of ammonium sulfate, 1 part of magnesium sulfate, 2 parts of dipotassium hydrogen phosphate, sulfur
Ferrous 0.01 part of acid, 20 parts of agar and 1000 parts of distilled water.
3. the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum as claimed in claim 1 preparation method it is characterised in that: step
2) mass ratio of described marine algae powder and water is 4:100；115 DEG C of sterilising temp, sterilization time 25min；During preparation seed liquor, huge
Bacterium anthracoides are seeded in fluid medium, cultivate 24 hours at 30 DEG C, obtain seed liquor, bacillus megaterium and culture medium
Volume ratio be 5:100；In step 3), seed liquor is 10:100 with the mass ratio of product a；In step 5) and step 7), temperature
Spend for 5 DEG C, the holding time is 12h.
4. the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum as claimed in claim 1 preparation method it is characterised in that: step
4), in step 6) and step 8), temperature is 30 DEG C, and ph is 6.5～7.0；Step 4) fermentation time is 8 days；Step 6) and step
8) holding time is 12h；The seed liquor supplemented in the mass ratio of the seed liquor and the product d that supplement in step 6) and step 8) with
The mass ratio of product f is 5:100.
5. the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum as claimed in claim 1 preparation method it is characterised in that: step
5), in step 6) step 7) and step 8), adjustment temperature is and adjusts 3 DEG C per hour.
6. the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum as claimed in claim 1 preparation method it is characterised in that: described
The mass ratio of the bacillus megaterium microbial inoculum of Chemical Mixed Fertilizer, parcel oil and cellulase-producing is 99.5:0.2:0.3.
7. the preparation method of the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum as described in any one of claim 1 to 6, its
It is characterised by: described Sargassum refers to the two or more of sargassum thunbergii, one of Sargassum phyllocystum Tseng et Lu,Sargassum horneri (Turn.) C. Ag. (Fucus horneri (Turn.)C.Ag.,Spongocarpus horneri Kutz.) and Alga Sgrgassi Enervess or arbitrary proportion.
8. the preparation method of the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum as described in any one of claim 1 to 6, its
It is characterised by: described bacillus megaterium is bacillus megaterium yic～bm1, preserving number is cgmcc no.12156.
9. the Chemical Mixed Fertilizer containing bacillus megaterium microbial inoculum as claimed in claim 7 preparation method it is characterised in that: described
Bacillus megaterium is bacillus megaterium yic～bm1, and preserving number is cgmcc no.12156.
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| CN108249987A (en) * | 2017-12-25 | 2018-07-06 | 云南云叶化肥股份有限公司 | A kind of compound fertilizer containing active microorganism and preparation method thereof |
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