CN102229900B - Compound probiotics preparation for biological organic fertilizers and preparation method thereof - Google Patents
Compound probiotics preparation for biological organic fertilizers and preparation method thereof Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The invention discloses a compound probiotics preparation for biological organic fertilizers and a preparation method thereof. The preparation method comprises the following steps: performing activated culture of 14 kinds of probiotics, including Pseudomona putida, Streptomyces violovariabilis, Rhizobium sp., Azotobacter chroococcum, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus cereus, Bacillus subtilis, Bacillus weihenstephanensis, Bacillus mucilaginosus, Bacillus thuringiensis, Saccharomyces rouxii, Streptomyces griseus var. ferrugineus, and Streptomyces microflavus; performing mixed inoculation at a proportion of 1% to 5% to a 500 mL liquid seed culture medium; then performing amplification culture; and finally delivering the strains to a 500 mL fermentor and fermenting for 3 to 5 days under such conditions that the temperature ranges from 28 DEG C to 30 DEG C and the rotational speed is 200 rpm, to obtain a liquid-state compound probiotics preparation. The compound probiotics preparation for biological organic fertilizers, provided by the invention, plays important roles of taking advantage of microbial ecology, improving soil, promoting plant growth, improving the yield and disease resistance of crops and reducing the consumption of fertilizers.
Description
Technical field
The present invention relates to biological organic fertilizer compound probiotic agent and preparation method thereof, belong to fields such as agricultural application, urban greening and environmental protection.
Background technology
Feces of livestock and poultry, agricultural crop straw, application of city life garbage and garden plant waste are the direct sources of disturbing world environments and health of people.Again because long-term irrational use chemical fertilizer; Cause part farmland nutritive ingredient out of proportion; Cause farmland ecological environment, soil physico-chemical characteristic and fauna soil microorganisms to receive destruction in various degree, injure the safety of agricultural-food and people's quality of life to a certain extent.And biological organic fertilizer not only provides plant-growth required nutrient, when replenishing soil fauna mikrobe, and the characteristics such as hardening, prevent salting of soil, minimizing fertilizer amount of can also effectively improving the soil.Therefore, biological organic fertilizer is playing an important role aspect realization world environments and the ecological Sustainable development.
In recent years, along with the prosperity of agrotechnical development and market economy, the biological organic fertilizer industry also develops rapidly, particularly aspect livestock excrement composting, has domesticly set up a lot of enterprises; Also started simultaneously with agricultural crop straw, application of city life garbage and garden plant waste is the upsurge of raw material matured compost research.At present compost fermentations that adopt under the natural condition or add decomposing microbial inoculum more.Under the natural condition, the fermentation period that becomes thoroughly decomposed is long, does not utilize industrialization production.Adding decomposing microbial inoculum can effectively shorten the fermentation maturity cycle, enhance productivity, but the product of producing does not meet the standard of biological organic fertilizer.Because the decomposing microbial inoculum and the soil microorganisms flora that add are inconsistent, so soil fauna microorganism species can not get replenishing; In addition, though some decomposing microbial inoculum contains the bacterial classification in the soil microorganisms flora, at present preceding production technique of adding probiotic agent of granulation that adopt because these beneficial microbe colonies are killed in high temperature granulation meeting more.Therefore, always do not reach the living bacteria count (>=0.2 hundred million/gram) that the biological organic fertilizer GB requires.
Summary of the invention
The purpose of this invention is to provide the agent of a kind of biological organic fertilizer compound probiotic.
Another object of the present invention provides the preparation method of a kind of biological organic fertilizer compound probiotic agent.
The preparation method of biological organic fertilizer compound probiotic of the present invention agent, be made up of the following step:
A, bacterial classification bring back to life
Adopt mikrobe: pseudomonas (Pseudomonas putida) GSICC 31605, purple variant streptomycete (Streptomyces violovariabilis) GSICC 41915, root nodule bacterium (Rhizobium sp.) GSICC 31813, aztobacter sp (Azotobacter chroococcum) GSICC 30112, subtilis (Bacillus subtilis) GSICC 32828, bacillus amyloliquefaciens (Bacillus amyloliquefaciens) GSICC 32826, bacillus cereus (Bacillus cereus) GSICC 32815, Bacillus licheniformis (Bacillus subtilis) GSICC 30285, Webster genus bacillus (Bacillus weihenstephanensis) GSICC 32844, bacillusmusilaginosiengineering (Bacillus mucilaginosus) GSICC 30250, bacillus thuringiensis (Bacillus thuringiensis) GSICC 32807, saccharomyces rouxii (Saccharomyces rouxii) GSICC 51930, streptomyces griseus rust mutation (Streptomyces griseus var. ferrugineus) GSICC 41920, streptomyces microflavus (treptomyces microflavus) GSICC 41903, more than 14 kinds of mikrobes all in Gansu Province Research for Industrial Microbial Germ preservation center (GSMSC) buy.
Because bacterial classification is preserved in 4 ℃ of strain libraries for a long time, therefore to bring back to life before use.Concrete rejuvenation method: get above-mentioned 14 kinds of mikrobes, after room temperature is placed 1-2 days, be transferred to respectively and be equipped with in the test tube that brings back to life substratum, cultivated 5 days, and just obtained activated spawn for 28-30 ℃.
Present method can be accomplished with existing bacteria culture medium, but effect is not best, and the weight proportion that best bacterial classification brings back to life substratum is: yam 20%, and glucose 2%, Carnis Bovis seu Bubali cream 0.3%, peptone 1%, sodium-chlor 0.5%, agar 1.5%, surplus is a water.The concrete making: get yam, remove the peel, be cut into piece and boil 30min, use 200 order filtered through gauze then, add above-mentioned other raw materials again, supply water, treat that agar dissolves back packing test tube, 121 ℃ of sterilization 30min.
B, combined inoculation and fermentation
The bacterial classification that activation is good adopts the combined inoculation method; According to pseudomonas: purple variant streptomycete: root nodule bacterium: aztobacter sp: subtilis: bacillus amyloliquefaciens: bacillus cereus: Bacillus licheniformis: Webster genus bacillus: bacillusmusilaginosiengineering: bacillus thuringiensis: saccharomyces rouxii: streptomyces griseus rust mutation: streptomyces microflavus is that the ratio of 1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5 is inoculated in the 500mL liquid seed culture medium; At 28-30 ℃; Cultivate on the shaking table that rotating speed is 200 rev/mins, fermented 2-4 days;
Enlarge fermentation then, during the fermentation, 14 kinds of mikrobes are regulated the flora ratio automatically, ferment with producing through seed fermentation, and the ratio of 14 kinds of microorganism species reaches best microecological balance state, best synergy between the performance mikrobe.At first bacterial classification is transferred in the 5L liquid seed culture medium, on the shaking table of 200 rev/mins of 28-30 ℃, rotating speed, cultivates, fermented 2-4 days; Then bacterial classification is transferred in the 50L liquid seed culture medium again, temperature 28-30 ℃, 200 rev/mins of rotating speeds fermented 2-4 days; At last bacterial classification is transported in the 500L fermentation tank culture medium, temperature 28-30 ℃, 200 rev/mins of rotating speeds fermented 3-5 days, can obtain the agent of liquid compound probiotic;
The weight proportion of 500mL, 5L, 50L liquid seed culture medium is: yam 20%, glucose 2%, Carnis Bovis seu Bubali cream 0.3%; Peptone 1%, sodium-chlor 0.5%, the concrete making: the yam of getting the 200g peeling; Be cut into piece and boil 30min; Use 200 order filtered through gauze then, add above-mentioned other raw materials again and supply water, 121 ℃ of sterilization 30min.
The weight proportion of 500L fermentation tank culture medium is: molasses 3-5%, and Carnis Bovis seu Bubali cream 0.3%, peptone 1%, sodium-chlor 0.5%, sal epsom 0.05%, potassium primary phosphate 0.05%, saltpetre 0.1% adds water and supplies 500L, and adjust pH is 6.8-7.4.
The preparation of C, the agent of solid compound probiotic
The weathered coal (available from jujube source, sky, Xinjiang Qiuci biotechnology Ltd) that B is obtained the agent of liquid compound probiotic and 5 tons mixes; Stacked 3-5 days; 55 ℃ of oven dry, promptly can be made into the solid-state version powder then, this solid compound probiotic agent viable count >=3,000,000,000/gram.
Just can make the agent of biological organic fertilizer compound probiotic with the inventive method, but finished product preservation of the present invention 6 months.
The method of use of microbial inoculum of the present invention: can cooperate uses such as biological compost, organic chemical fertilizer and compound fertilizer, can make base fertilizer, also can do and append fertilizer.Microbial inoculum and composite fertilizer are according to the 1:100 mass ratio, and after mixing, the base fertilizer dose is about 30-40kg/ mu, append fertile dose and are about 5-10kg/ mu.Also can be after composite fertile production, according to adding according to the 1:100 mass ratio, packing then, biological organic fertilizer.This 6 months biological organic fertilizer quality guaranteed perioves, to make base fertilizer and use, dose is about 30-40kg/ mu, appends fertile dose and is about 5-10kg/ mu.
The useful microbe bacterial classification that the present invention adds has following function:
1) mikrobes such as aztobacter sp, root nodule bacterium, the mutation of streptomyces griseus rust have fixedly nitrogen ability, can reduce the consumption of chemical nitrogen fertilizer;
2) mikrobes such as Webster genus bacillus, the mutation of streptomyces griseus rust, pseudomonas, pesticide residue and water pollutant in the soil of degrading;
3) mikrobes such as purple variant streptomycete, bacillusmusilaginosiengineering, the mutation of streptomyces griseus rust can be decomposed the organophosphorus in the soil, transform invalid phosphorus, potassium etc., help the absorption of plant to element;
4) mikrobe such as bacillus cereus, bacillusmusilaginosiengineering, the silicate in the soil of degrading, reduction nitrate salt improves problems such as soil compaction;
5) mikrobe such as bacillus amyloliquefaciens, saccharomyces rouxii can tolerate salt affected soil, and can the saline and alkaline composition of metabolism, thus the saline and alkaline of improving the soil;
6) mikrobes such as streptomyces microflavus, the mutation of streptomyces griseus rust, purple variant streptomycete can secrete the required hormone of plant, promote plant-growth, solid, improve output;
7) mikrobes such as streptomyces microflavus, the mutation of streptomyces griseus rust, subtilis, bacillus amyloliquefaciens, bacillus thuringiensis can secrete some microbiotic or produce gemma, can improve the disease resistance of crop, reduce pesticide dosage.
Compound probiotic agent of the present invention contains 14 kinds of effective strains, through combined inoculation, makes it regulate various mikrobe ratios automatically, has reached best little equilibrium state.Under this state, their synergies are the strongest.
The present invention is according to the soil microorganisms ecosystem characterization, and supply soil shortage beneficial microbe colony cooperates biological compost to use targetedly; Make the biological organic fertilizer living bacteria count reach the GB requirement; Compound probiotic agent of the present invention can be improved the soil, improves soil depletion, hardened, and degrading pesticide pollutes, and prevents salting of soil; Promote plant-growth, raising crop yield and aspects such as disease resistance, minimizing fertilizer amount to play a significant role; The preparation method is simple simultaneously, and easy handling is fit to scale operation and application.
Embodiment
Following embodiment can further specify the present invention, but does not limit the present invention in any way.
Before practical implementation, at first Research for Industrial Microbial Germ preservation center (GSMSC) obtains the inclined-plane of 14 bacterial classifications from Gansu Province.Because bacterial classification is preserved in 4 ℃ of strain libraries for a long time, therefore to bring back to life before use.Concrete rejuvenation method: get above bacterial classification from the preservation center, after room temperature is placed 1-2 days, be transferred to the test tube slant respectively, cultivated 5 days, and just obtained activated spawn for 28-30 ℃.
The 1L bacterial classification brings back to life culture medium prescription: yam 200g, glucose 20g, Carnis Bovis seu Bubali cream 3g, peptone 10g, sodium-chlor 5g, agar 15g.The concrete making: get the yam of 200g peeling, be cut into piece and boil 30min, use 200 order filtered through gauze then, add above-mentioned other raw materials again, treat to supply water to 1L after agar dissolves, packing test tube, 121 ℃ of sterilization 30min.
Embodiment 1
The bacterial classification that activation is good adopts the combined inoculation method; According to pseudomonas: purple variant streptomycete: root nodule bacterium: aztobacter sp: subtilis: bacillus amyloliquefaciens: bacillus cereus: Bacillus licheniformis: Webster genus bacillus: bacillusmusilaginosiengineering: bacillus thuringiensis: saccharomyces rouxii: streptomyces griseus rust mutation: the mixed of streptomyces microflavus=1:1:1:1:1:1:1:1:1:1:1:1:1:1 is inoculated in the 500mL liquid seed culture medium; On the shaking table of 200 rev/mins of 28 ℃, rotating speed, cultivate, fermented 2 days.Be transferred to the 5L secondary seed then and shake in the bottle, on the shaking table of 200 rev/mins of 28 ℃, rotating speed, cultivate, fermented 2 days.Be transferred to again in the 50L seeding tank, 28 ℃ of temperature, 200 rev/mins of rotating speeds fermented 2 days.The bacterial classification of seeding tank is transported in the 500L fermentor tank, 28 ℃ of temperature, 200 rev/mins of rotating speeds fermented 3 days, obtained the agent of liquid compound probiotic.
The weathered coal that obtains the agent of liquid compound probiotic and 5 tons is mixed, stacked 3 days, 55 ℃ of oven dry, promptly can be made into the solid-state version powder then, this solid compound probiotic agent living bacteria count is 3,000,000,000 a/gram.
5OOmL liquid seed culture medium (first order seed shake-flask culture base): yam 100g, glucose 10g, Carnis Bovis seu Bubali cream 1.5g, peptone 5g, sodium-chlor 2.5g.The concrete making: get the yam of 100g peeling, be cut into piece and boil 30min, use 200 order filtered through gauze then, add above-mentioned other raw materials again and supply water, 121 ℃ of sterilization 30min.
5L liquid seed culture medium prescription (secondary seed shake-flask culture base): yam 1000g, glucose 100g, Carnis Bovis seu Bubali cream 15g, peptone 50g, sodium-chlor 25g.The concrete making: get the yam of 1000g peeling, be cut into piece and boil 30min, use 200 order filtered through gauze then, add above-mentioned other raw materials again and supply water, 121 ℃ of sterilization 30min.
50L liquid seed culture medium prescription (seed tank culture base): yam 10000g, glucose 1000g, Carnis Bovis seu Bubali cream 150g, peptone 500g, sodium-chlor 250g.The concrete making: get the yam of 10000g peeling, be cut into piece and boil 30min, use 200 order filtered through gauze then, add above-mentioned other raw materials again and supply water, in seeding tank, sterilize.
The 500L fermentation tank culture medium is formed and mass content is: molasses 3%, i.e. 15000g; Carnis Bovis seu Bubali cream 0.3%, i.e. 1500g; Peptone 1%, i.e. 5000g; Sodium-chlor 0.5%, i.e. 2500g; Sal epsom 0.05%, i.e. 250g; Potassium primary phosphate 0.05%, i.e. 250g; Saltpetre 0.1%, i.e. 500g; Add water and supply 500L, regulating the pH value is 6.8.
Embodiment 2
Change fermentation time and temperature among the embodiment 1, promptly the leavening temperature of level liquid seed, secondary liquid seeds, seeding tank and enlarged culturing has 28 ℃ to change into 30 ℃, and fermentation time had 2 days to be extended for 3 days, simultaneously molasses concentration was increased to 5% from 3%.The weathered coal that obtains the agent of liquid compound probiotic and 5 tons is mixed, and pilling up time also had 3 days to be extended for 5 days.Process the solid-state version powder under this condition, this solid compound probiotic agent living bacteria count is 3,800,000,000 a/gram.
The liquid seed culture medium prescription is identical with embodiment 1.
The 500L fermentation tank culture medium is formed and mass content is: molasses 5%, i.e. 25000g; Carnis Bovis seu Bubali cream 0.3%, i.e. 1500g; Peptone 1%, i.e. 5000g; Sodium-chlor 0.5%, i.e. 2500g; Sal epsom 0.05%, i.e. 250g; Potassium primary phosphate 0.05%, i.e. 250g; Saltpetre 0.1%, i.e. 500g; Add water and supply 500L, regulating the pH value is 6.8.
Embodiment 3
The pH6.8 that changes fermention medium among the embodiment 1 is pH7.4, and leavening temperature is identical with embodiment 2 with fermentation time.Promptly; According to pseudomonas: purple variant streptomycete: root nodule bacterium: aztobacter sp: subtilis: bacillus amyloliquefaciens: bacillus cereus: Bacillus licheniformis: Webster genus bacillus: bacillusmusilaginosiengineering: bacillus thuringiensis: saccharomyces rouxii: the ratio inoculation of streptomyces griseus rust mutation: streptomyces microflavus=1:1:1:1:1:1:1:1:1:1:1:1:1:1; The leavening temperature of level liquid seed, secondary liquid seeds, seeding tank and enlarged culturing is 30 ℃; Fermentation time is 3 days; The pH value is 7.4, molasses concentration 3%, pilling up time 5 days.Process the solid-state version powder under this condition, this solid compound probiotic agent living bacteria count is 3,500,000,000 a/gram.
The liquid seed culture medium prescription is identical with embodiment 1.
The 500L fermentation tank culture medium is formed and mass content is: molasses 3%, i.e. 15000g; Carnis Bovis seu Bubali cream 0.3%, i.e. 1500g; Peptone 1%, i.e. 5000g; Sodium-chlor 0.5%, i.e. 2500g; Sal epsom 0.05%, i.e. 250g; Potassium primary phosphate 0.05%, i.e. 250g; Saltpetre 0.1%, i.e. 500g; Add water and supply 500L, regulating the pH value is 7.4.
Embodiment 4
Molasses concentration is 5% among the change embodiment 3; And according to pseudomonas: purple variant streptomycete: root nodule bacterium: aztobacter sp: subtilis: bacillus amyloliquefaciens: bacillus cereus: Bacillus licheniformis: Webster genus bacillus: bacillusmusilaginosiengineering: bacillus thuringiensis: saccharomyces rouxii: the ratio inoculation of streptomyces griseus rust mutation: streptomyces microflavus=1:1:1:1:1:1:1:1:1:1:1:1:1:1; The leavening temperature of level liquid seed, secondary liquid seeds, seeding tank and enlarged culturing is 28 ℃; Fermentation time is 3 days; The pH value is 7.4, molasses concentration 5%, pilling up time 5 days.Process the solid-state version powder under this condition, this solid compound probiotic agent living bacteria count is 3,400,000,000 a/gram.
The liquid seed culture medium prescription is identical with embodiment 1.
The 500L fermentation tank culture medium is formed and mass content is: molasses 5%, i.e. 25000g; Carnis Bovis seu Bubali cream 0.3%, i.e. 1500g; Peptone 1%, i.e. 5000g; Sodium-chlor 0.5%, i.e. 2500g; Sal epsom 0.05%, i.e. 250g; Potassium primary phosphate 0.05%, i.e. 250g; Saltpetre 0.1%, i.e. 500g; Add water and supply 500L, regulating the pH value is 7.4.
Embodiment 5
Regulate inoculative proportion; According to pseudomonas: purple variant streptomycete: root nodule bacterium: aztobacter sp (Azotobacter chroococcum): subtilis: bacillus amyloliquefaciens (Bacillus amyloliquefaciens): bacillus cereus: Bacillus licheniformis: Webster genus bacillus: bacillusmusilaginosiengineering: bacillus thuringiensis: saccharomyces rouxii: the mutation of streptomyces griseus rust: the ratio of streptomyces microflavus=5:5:5:5:5:5:5:5:5:5:5:5:5:5 is inoculated; Other fermentation condition is identical with embodiment 2; Promptly; The leavening temperature of level liquid seed, secondary liquid seeds, seeding tank and enlarged culturing is 30 ℃, and fermentation time is 3 days, and the pH value is 6.8; Molasses concentration 5%, pilling up time 5 days.Process the solid-state version powder under this condition, this solid compound probiotic agent living bacteria count is 3,700,000,000 a/gram.
The liquid seed culture medium prescription is identical with embodiment 1.
The 500L fermentation tank culture medium is formed and mass content is: molasses 5%, i.e. 25000g; Carnis Bovis seu Bubali cream 0.3%, i.e. 1500g; Peptone 1%, i.e. 5000g; Sodium-chlor 0.5%, i.e. 2500g; Sal epsom 0.05%, i.e. 250g; Potassium primary phosphate 0.05%, i.e. 250g; Saltpetre 0.1%, i.e. 500g; Add water and supply 500L, regulating the pH value is 6.8.
Embodiment 6
The pH that changes embodiment 5 fermentation culture collection is 7.4; Other condition is all identical with embodiment 5; Promptly according to pseudomonas: purple variant streptomycete: root nodule bacterium: aztobacter sp (Azotobacter chroococcum): subtilis: bacillus amyloliquefaciens (Bacillus amyloliquefaciens): bacillus cereus: Bacillus licheniformis: Webster genus bacillus: bacillusmusilaginosiengineering: bacillus thuringiensis: saccharomyces rouxii: the mutation of streptomyces griseus rust: the ratio of streptomyces microflavus=5:5:5:5:5:5:5:5:5:5:5:5:5:5 is inoculated; The leavening temperature of level liquid seed, secondary liquid seeds, seeding tank and enlarged culturing is 30 ℃; Fermentation time is 3 days; The pH value is 7.4, molasses concentration 5%, pilling up time 5 days.Process the solid-state version powder under this condition, this solid compound probiotic agent living bacteria count is 37.5 hundred million a/gram.
The liquid seed culture medium prescription is identical with embodiment 1.
The 500L fermentation tank culture medium is formed and mass content is: molasses 5%, i.e. 25000g; Carnis Bovis seu Bubali cream 0.3%, i.e. 1500g; Peptone 1%, i.e. 5000g; Sodium-chlor 0.5%, i.e. 2500g; Sal epsom 0.05%, i.e. 250g; Potassium primary phosphate 0.05%, i.e. 250g; Saltpetre 0.1%, i.e. 500g; Add water and supply 500L, regulating the pH value is 7.4.
Embodiment 7
Inoculum size is identical with embodiment 5; Other fermentation condition is identical with embodiment 1; Promptly according to pseudomonas: purple variant streptomycete: root nodule bacterium: aztobacter sp (Azotobacter chroococcum): subtilis: bacillus amyloliquefaciens (Bacillus amyloliquefaciens): bacillus cereus: Bacillus licheniformis: Webster genus bacillus: bacillusmusilaginosiengineering: bacillus thuringiensis: saccharomyces rouxii: the mutation of streptomyces griseus rust: the ratio of streptomyces microflavus=5:5:5:5:5:5:5:5:5:5:5:5:5:5 is inoculated; The leavening temperature of level liquid seed, secondary liquid seeds, seeding tank and enlarged culturing is 28 ℃; Fermentation time is 2 days; The pH value is 6.8, molasses concentration 3%, pilling up time 3 days.Process the solid-state version powder under this condition, this solid compound probiotic agent living bacteria count is 3,100,000,000 a/gram.
The liquid seed culture medium prescription is identical with embodiment 1.
The 500L fermentation tank culture medium is formed and mass content is: molasses 3%, i.e. 15000g; Carnis Bovis seu Bubali cream 0.3%, i.e. 1500g; Peptone 1%, i.e. 5000g; Sodium-chlor 0.5%, i.e. 2500g; Sal epsom 0.05%, i.e. 250g; Potassium primary phosphate 0.05%, i.e. 250g; Saltpetre 0.1%, i.e. 500g; Add water and supply 500L, regulating the pH value is 6.8.
Embodiment 8
Fermention medium is identical with embodiment 6, and fermentation condition is identical with embodiment 1.Promptly according to pseudomonas: purple variant streptomycete: root nodule bacterium: aztobacter sp (Azotobacter chroococcum): subtilis: bacillus amyloliquefaciens (Bacillus amyloliquefaciens): bacillus cereus: Bacillus licheniformis: Webster genus bacillus: bacillusmusilaginosiengineering: bacillus thuringiensis: saccharomyces rouxii: the mutation of streptomyces griseus rust: the ratio of streptomyces microflavus=5:5:5:5:5:5:5:5:5:5:5:5:5:5 is inoculated; The leavening temperature of level liquid seed, secondary liquid seeds, seeding tank and enlarged culturing is 28 ℃; Fermentation time is 2 days; The pH value is 7.4; Molasses concentration 5%, pilling up time 3 days.Process the solid-state version powder under this condition, this solid compound probiotic agent living bacteria count is 3,200,000,000 a/gram.
The liquid seed culture medium prescription is identical with embodiment 1.
The 500L fermentation tank culture medium is formed and mass content is: molasses 5%, i.e. 25000g; Carnis Bovis seu Bubali cream 0.3%, i.e. 1500g; Peptone 1%, i.e. 5000g; Sodium-chlor 0.5%, i.e. 2500g; Sal epsom 0.05%, i.e. 250g; Potassium primary phosphate 0.05%, i.e. 250g; Saltpetre 0.1%, i.e. 500g; Add water and supply 500L, regulating the pH value is 7.4.
Embodiment 9
Culture condition is identical with embodiment 2, and inoculative proportion is different.Promptly according to pseudomonas: purple variant streptomycete: root nodule bacterium: aztobacter sp (Azotobacter chroococcum): subtilis: bacillus amyloliquefaciens (Bacillus amyloliquefaciens): bacillus cereus: Bacillus licheniformis: Webster genus bacillus: bacillusmusilaginosiengineering: bacillus thuringiensis: saccharomyces rouxii: the mutation of streptomyces griseus rust: the ratio of streptomyces microflavus=1:1:1:2:2:2:3:3:3:4:4:4:5:5 is inoculated; The leavening temperature of level liquid seed, secondary liquid seeds, seeding tank and enlarged culturing is 30 ℃; Fermentation time is 3 days; The pH value is 6.8; Molasses concentration 5%, pilling up time 5 days.Process the solid-state version powder under this condition, this solid compound probiotic agent living bacteria count is 3,800,000,000 a/gram.
The liquid seed culture medium prescription is identical with embodiment 1.
The 500L fermentation tank culture medium is formed and mass content is: molasses 5%, i.e. 25000g; Carnis Bovis seu Bubali cream 0.3%, i.e. 1500g; Peptone 1%, i.e. 5000g; Sodium-chlor 0.5%, i.e. 2500g; Sal epsom 0.05%, i.e. 250g; Potassium primary phosphate 0.05%, i.e. 250g; Saltpetre 0.1%, i.e. 500g; Add water and supply 500L, regulating the pH value is 6.8.
The experimental result of above embodiment shows:
The variation of 1, combined inoculation amount and pH value, little to the influence of the total living bacteria count of compound probiotic agent.
2, the concentration of fermentation time and fermention medium molasses is bigger to the influence of the total living bacteria count of compound probiotic agent.
3, leavening temperature has certain influence to the total living bacteria count of compound probiotic agent.
4, in condition of the present invention, the total living bacteria count of compound probiotic agent >=3,000,000,000/gram.
Following examples are that the compound probiotic agent that embodiment 1 obtains is mixed result of use with cow dung compost.
Practical example 1: the effect of compound probiotic agent on cucumber
Microbial inoculum and cow dung compost mix according to the 1:100 mass ratio, and plastic greenhouse cucumber base fertilizer dose is a 35kg/ mu.Contrast is cow dung compost, and plastic greenhouse cucumber base fertilizer dose is a 35kg/ mu.In cucumber growth, solid process, observe plant strain growth, poisoning situation, analyze white enzymophathy, the blight state of an illness, calculate disease index, protection effect and effect of increasing production.The result shows: test group cucumber frost enzymophathy disease refers to 15.8%, and control group cucumber frost enzymophathy disease refers to 25%, and the state of an illness alleviates 36.8%; Test group cucumber fusarium axysporum diseased plant rate 8%, control group cucumber fusarium axysporum diseased plant rate 20.5%, the state of an illness alleviates 60.9%.In addition, test the group cucumber and shift to an earlier date 5 days maturations than control group.Aspect the raising cucumber yield, test group obviously is superior to control group, test group output 5500kg/ mu, and control group output 4548kg/ mu is tested group and is increased income 20.9% than control group.
Practical example 2: the effect of compound probiotic agent on corn
Microbial inoculum and cow dung compost mix according to the 1:100 mass ratio.Carry out the fertilization effect test in corn test base, the sky and water, Gansu Province, the base fertilizer dose is a 40kg/ mu.Contrast is cow dung compost, and the base fertilizer dose is a 40kg/ mu.The interpolation of topdressing boot stage: the composite fertile dose of agent of experimental group compound probiotic and cow dung compost is a 8kg/ mu, contrasts to be that cow dung compost, dose are 8kg/ mu.Analyze and get 10 strain corns at random, analyze it and grow, measurement index: leaf area index (LAI) and plant height (PH); Corn yield, measurement index: spike length, tassel row number, 100-grain weight etc., the result sees table 1 and table 2.The result shows that experimental group each item index all is superior to control group.
Table 1: Different Fertilization is to the influence of leaf of Semen Maydis area index and plant height
Table 2: Different Fertilization is to the influence of corn yield index
| Project | Spike length/cm | Fringe is thick/cm | Tassel row number/individual | Row grain number/individual | 100-grain weight/g |
| Experimental group | 20.6 | 5.1 | 15.8 | 48 | 27.8 |
| Control group | 19.3 | 5.1 | 14.5 | 47 | 26.5 |
Practical example 3: the effect of compound probiotic agent on ivy
Microbial inoculum and cow dung compost mix according to the 1:100 mass ratio.Spring in 2010, garden gardening greening development dawn in the Xu Jia mountain spring fertilization effect test in the Lanzhou City, Gansu Province.Ivy root dose is the 0.5kg/ strain.Contrast is cow dung compost, and ivy root dose is the 0.5kg/ strain.Late summer in 2010 was analyzed the ivy upgrowth situation, and the result sees 3.It is thus clear that the ivy of experimental group is flourishing, growth conditions is superior to control group.
Table 3: Different Fertilization is to the influence of ivy growth
| Project | Growing height/cm | Individual plant area coverage/cm 2 | Branch capability | The leaf look |
| Experimental group | 185 | 2.54 | By force | Blackish green |
| Control group | 126 | 1.72 | Medium | Green |
Above compound probiotic agent effect test result of the present invention shows:
1, after the interpolation microbial inoculum of the present invention, can obviously improve the crop disease-resistant ability, the effect of increasing income is remarkable.
2, this microbial inoculum is applied widely, can be used for aspects such as farm crop, melon and fruit, vegetables and afforestation.
3, this microbial inoculum use is simple to operate, promote the wider application easily.
Claims (5)
1. the preparation method of biological organic fertilizer compound probiotic agent is characterized in that it is made up of the following step:
A, bacterial classification bring back to life
Adopt 14 kinds of mikrobes; Be pseudomonas (Pseudomonas putida) GSICC 31605; Purple variant streptomycete (Streptomyces violovariabilis) GSICC 41915; Root nodule bacterium (Rhizobium sp.) GSICC 31813; Aztobacter sp (Azotobacter chroococcum) GSICC 30112; Subtilis (Bacillus subtilis) GSICC 32828; Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) GSICC 32826; Bacillus cereus (Bacillus cereus) GSICC 32815; Bacillus licheniformis (Bacillus subtilis) GSICC 30285; Webster genus bacillus (Bacillus weihenstephanensis) GSICC 32844; Bacillusmusilaginosiengineering (Bacillus mucilaginosus) GSICC 30250; Bacillus thuringiensis (Bacillus thuringiensis) GSICC 32807; Saccharomyces rouxii (Saccharomyces rouxii) GSICC 51930; Streptomyces griseus rust mutation (Streptomyces griseus var. ferrugineus) GSICC 41920; Streptomyces microflavus (treptomyces microflavus) GSICC 41903
Get above-mentioned 14 bacterial classifications, after room temperature is placed 1-2 days, be transferred to 14 respectively again and be equipped with in the test tube that brings back to life substratum, cultivated 5 days, and just obtained activated spawn for 28-30 ℃;
B, combined inoculation and fermentation
The bacterial classification that activation is good adopts the combined inoculation method; According to pseudomonas: purple variant streptomycete: root nodule bacterium: aztobacter sp: subtilis: bacillus amyloliquefaciens: bacillus cereus: Bacillus licheniformis: Webster genus bacillus: bacillusmusilaginosiengineering: bacillus thuringiensis: saccharomyces rouxii: streptomyces griseus rust mutation: streptomyces microflavus is that the ratio of 1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5:1-5 is inoculated in the 500mL liquid seed culture medium; At 28-30 ℃; Cultivate on the shaking table that rotating speed is 200 rev/mins, fermented 2-4 days; Be transferred to then in the 5L liquid seed culture medium, on the shaking table of 200 rev/mins of 28-30 ℃, rotating speed, cultivate, fermented 2-4 days; Be transferred in the 50L liquid seed culture medium, temperature 28-30 ℃, 200 rev/mins of rotating speeds fermented 2-4 days again; The bacterial classification of seeding tank is transported in the 500L fermentation tank culture medium, and temperature 28-30 ℃, 200 rev/mins of rotating speeds fermented 3-5 days, can obtain the agent of liquid compound probiotic;
The preparation of C, the agent of solid compound probiotic
The weathered coal that the B step is obtained the agent of liquid compound probiotic and 5 tons mixes, and stacks 3-5 days, 55 ℃ of oven dry, promptly can be made into the solid-state version powder then.
2. the preparation method of biological organic fertilizer compound probiotic according to claim 1 agent; The weight proportion that it is characterized in that bacterial classification resurrection substratum in the said steps A is: yam 20%, glucose 2%, Carnis Bovis seu Bubali cream 0.3%, peptone 1%, sodium-chlor 0.5%, agar 1.5%, surplus are water.
3. the preparation method of biological organic fertilizer compound probiotic according to claim 1 agent; The weight proportion that it is characterized in that liquid seed culture medium among the said step B is: yam 20%, glucose 2%, Carnis Bovis seu Bubali cream 0.3%, peptone 1%, sodium-chlor 0.5%, surplus are water.
4. the preparation method of biological organic fertilizer compound probiotic according to claim 1 agent; The weight proportion that it is characterized in that fermentation tank culture medium among the said step B is: molasses 3-5%, Carnis Bovis seu Bubali cream 0.3%, peptone 1%, sodium-chlor 0.5%, sal epsom 0.05%, potassium primary phosphate 0.05%, saltpetre 0.1%, surplus are water.
5. the biological organic fertilizer compound probiotic agent that makes according to any one said preparation method among the claim 2-4.
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