CN103553828A - Liquid compound biological fertilizer with long shelf life as well as preparation method and application thereof - Google Patents
Liquid compound biological fertilizer with long shelf life as well as preparation method and application thereof Download PDFInfo
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- CN103553828A CN103553828A CN201310561591.5A CN201310561591A CN103553828A CN 103553828 A CN103553828 A CN 103553828A CN 201310561591 A CN201310561591 A CN 201310561591A CN 103553828 A CN103553828 A CN 103553828A
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Abstract
The invention relates to a liquid compound biological fertilizer with long shelf life as well as a preparation method and application thereof. The liquid compound biological bacterial fertilizer comprises the following components in parts by weight: 2-5 parts of microbial thalli, 80-130 parts of compound nutrients, 10-30 parts of thalli protective agents and 0.5-3.0 parts of additives, wherein the microbial thalli is metered in terms of the wet weight of deposition obtained after fermentation liquor is centrifugalized, the microbial thalli include bacillus thuringiensis, bacillus megaterium, bacillus mucilaginosus and bacillus amyloliquefaciens, and the total viable count achieves 5*10<8>-5*10<9>/mL. The liquid compound biological bacterial fertilizer disclosed by the invention can be used as a biological pesticide, can be directly applied to the spraying of plant surfaces or applied to seed soaking, root irrigation, and the like, has the effects of insect resistance, phosphate solubilization, potassium solubilization, organic matter solubilization, pest resistance, and the like and is long in shelf life.
Description
Technical field
The invention belongs to composite microbic bacterial fertilizer field, be specifically related to a kind of liquid composite bio-fertilizer of growing the quality guaranteed period and its preparation method and application.
Background technology
Conservation culture is contemporary trend, and the development of agricultural is closely bound up with ecotope, healthy ecotope can provide strong support to guarantee to agricultural, and can be important foundation and the guarantee of agricultural development, but owing to using for a long time chemical fertilizer or organic fertilizer, and Fertilization Structure unreasonable causes the situation of pollution that China is faced with agroecological environment and destruction, soil physical chemistry degradation and caused many serious problems.In green agriculture, become today of world agriculture developing direction, we should advocate, develop and use microbial fertilizer energetically, adhere to walking the road of Sustainable development.
Micro organism composite fertilizer, is to take beneficial microorganism as main, is equipped with a kind of novel green fertilizer that organic and inorganic composition forms.The vital movement of microorganism in composite fertilizer, nutrient in can activating soil, thereby improve the supply of plant nutrient, vital movement by microorganism produces plant growth substance, stimulating plant growth, promotes the absorption of crop to nutritive element, some pathogenic microorganism of antagonism, alleviate crop pest, and increase output.
Bacillus thuringiensis (Bacillus thuringiensis), can produce specific killing action to insect, respectively the various insects such as lepidopteran, Diptera, Coleoptera, Hymenoptera, Homoptera, Orthoptera, Mallophaga is had to activity.Bacillus megaterium (Bacillusmegaterium), can decompose insoluble phosphorus compound, and has stronger nitrite degradation ability.Colloid bacillus cereus (Bacillus mucilaginosus), can decompose the ecosystem mineral of the aluminum silicates such as feldspar, mica, makes insoluble K, P, Si etc. in soil change solubility into for plant-growth utilization.Can also produce the biologically active substance stimulating plant growths such as growth hormone, phytokinin.It is reported that colloid bacillus cereus has volume increase, improves quality, improves the effects such as crop anti-adversity ability various crop.In pure culture, also find that silicate bacteria can produce multiple amino acids, peptides simultaneously, and the physiologically active substance such as growth hormone, Plant hormones regulators,gibberellins, the phytopathogens such as cotton wilt, tobacco gray mold, wheat scab are had to obvious restraining effect, can alleviate to a certain extent the particularly generation of some soil-borne diseases of certain plants disease.Bacillus amyloliquefaciens (Bacillus amyloliquefaciens); organic matter decomposition ability is strong; meta-bolites is abundant; producing multiplely has inhibiting material to plant pathogenic fungi; simultaneously higher to the restraint of spoilage organism and pathogenic bacteria, reproduction speed is fast, forms poly-Vetsin; form soil protection film, moisture retention is strong.
The composite microbic bacterial fertilizer of selling is in the market more and more, but most bacterial manure effect is comparatively single, and price is high, and most of liquid bacterial manure quality guaranteed period is short, the easy inactivation of thalline.Liquid composite biological fertilizer of the present invention, adopt the mode of production cheaply to produce, thereby select function stem to make product integrate pest-resistant, phosphorus decomposing, potassium decomposing, the organic effect that reaches crop yield of solution, and added function salt, can be used as trace element and provide nutrition for plant, also can make product reach the system of high osmotic pressure, contribute to improve yield, the stability of gemma, improve cortex construction, and increase its heat resist power.Meanwhile, added food sanitas, the vegetative activity of thalline in inhibition system, reaches the longer quality guaranteed period.
In marine alga, peculiar Sargassum polysaccharides, alginic acid, highly unsaturated fatty acids and multiple natural phant growth regulator, have very high biological activity, can stimulating plant body in the generation of non-specific active factor, regulate Endogenous hormone balance.In marine alga, contain natural phant growth regulator as growth hormone, cytokinin-like substance and Plant hormones regulators,gibberellins etc., have very high biological activity, in the ratio of various materials and terrestrial plant, each hormone ratio is close.Lalgine in marine alga can reduce the surface tension of water, at plant surface, form thin film, enlarge active surface, make water-soluble substances ratio be easier to see through cauline leaf surface detail after birth and enter vegetable cell, make plant absorb most effectively the nutritive ingredient in seaweed extracted liquor, if therefore liquid seaweed manure and sterilant, sterilant and chemical fertilizer mixing use, effect is better, can reduce sprinkling expense, agricultural chemicals and chemical fertilizer are had to synergism.Marine alga can directly make soil or by plant, soil be increased organic, activates the various beneficial microorganisms in soil, and these biologies can play a part catalyzer in the circulation of plant-microbe metabolite, and the biopotency of soil is increased.The metabolite of plants and soil microorganism can be plant more nutrient is provided.Marine alga can be worked harmoniously with plant-soil ecosystem.The natural compounds that it contains as Sodium Alginate be natural soil amendment, can promote the formation of soil aggregate, improve soil internal void space, coordinate solid, liquid, gas three ratio in soil, recovery, due to the soil lac balance that over-burden and chemical pollution loses, increases soil organisms vigor, promotes the release of available nutrient, be conducive to root growth, improve the resistance of crop.
Summary of the invention
The object of this invention is to provide a kind ofly have that pest-resistant, phosphorus decomposing, potassium decomposing, solution are organic, liquid composite biological fertilizer and the production method thereof of pest-resistant harmful effect and long quality-guarantee period.
To achieve these goals, the present invention adopts following technical scheme:
The composition of described liquid composite biological fertilizer comprises microbial cells, compound nutrients, thalline protective material and additive; Its parts by weight consist of, microbial cells 2-5, and compound nutrients 80-130, thalline protective material 10-30, additive 0.5-3.0, wherein microbial cells is the precipitation weight in wet base after centrifugal in fermented liquid.
The composition of described microbial cells comprises: bacillus thuringiensis (Bacillus thuringiensis), bacillus megaterium (Bacillusmegaterium), colloid bacillus cereus (Bacillus mucilaginosus), bacillus amyloliquefaciens (Bacillus amylolquefaciens), wherein total viable count reaches 5 * 10
8-5 * 10
9individual/mL.
Preferably, in microbial cells, four kinds of bacterium viable count ratios are as follows: bacillus thuringiensis: bacillus megaterium: colloid bacillus cereus: bacillus amyloliquefaciens=1-2:2-3:2-3:1-2.
The parts by weight of described compound nutrients consist of:
Potassium primary phosphate 50-150,
EDTA-Ca25-30，
EDTA-Mg or EDTA-Mg and MgSO
47H
2the two mixture 50-80 of O,
EDTA-Zn or EDTA-Zn and ZnSO
47H
2o or the two mixture 0.15-0.18,
EDTA-Mn or EDTA-Mn and MnSO
4h
2o or the two mixture 0.3-0.5,
EDTA-Fe0.1-0.3，
EDTA-Cu0.06-0.08，
Boric acid 0.1-0.3,
Humic acids 5-15,
Soybean protein 20-100,
Aminoacids complex 50,
Seaweed powder 20-100;
The parts by weight of described aminoacids complex consist of: L-Ala 6.0-6.5, arginine 6.5-6.8, aspartic acid 6.8-7.2, Gelucystine 2.2-2.5, glycine 3.2-3.5, Histidine 2.2-2.5, Methionin 6.4-6.5, methionine(Met) 3.0-3.2, phenylalanine 5-5.5, proline-4-4.2, Serine 8.4-8.5, Threonine 4.8-5.0, tryptophane 1.5-1.8, trorsine 14 .2-4.5, α-amino-isovaleric acid 7.4-7.5;
Described thalline protective material is one or more in cell immobilization carrier, includes but not limited to diatomite, kaolin, talcum powder etc.;
Described additive is one or more in organic acid, includes but not limited to oxysuccinic acid, citric acid, lactic acid etc.
The production method of described liquid composite biological fertilizer is as follows:
(1) actication of culture and seed liquor preparation
Bacillus thuringiensis, bacillus megaterium, colloid bacillus cereus, Bacillus amyloliquefaciens strain are activated respectively and cultivated and make seed liquor;
(2) fermentation culture
The seed liquor of cultured bacillus thuringiensis, bacillus megaterium, bacillus amyloliquefaciens is seeded to 5L fermentor tank with the inoculum size of 8%-10% respectively, fermention medium is substratum No. 3, and 37 ℃～40 ℃, rotating speed 300rpm, ventilating ratio 1:1, collects fermented liquid after fermentation 15-18h;
The seed liquor of cultured colloid bacillus cereus is seeded to 5L fermentor tank with the inoculum size of 8%-10%, and fermention medium is No. 4 substratum, 37 ℃～40 ℃, and rotating speed 400rpm, ventilating ratio 1:1, collects fermented liquid after fermentation 15-18h;
Described No. 3 substratum form: by percentage to the quality, and 1.0%-1.2% glucose, 0.5%-0.6%K
2hPO
43H
2o, 0.3%-0.5%KH
2pO
4, 0.1%-0.2% urea, 0.4%-0.6% yeast powder, 0.35%-0.45 ‰ MgSO
47H
2o, all the other are water, pH7.0-7.2;
Described No. 4 substratum: by percentage to the quality, 1% sucrose, 0.5%K
2hPO
43H
2o, 0.5 ‰ yeast powders, 0.02 ‰ FeCl
36H
2o, 0.4 ‰ CaCO
3, 0.2 ‰ MgSO
47H
2o, all the other are water, pH7.0;
(3) microorganism collection is with fixing
Four kinds of fermented liquids that step (2) is collected mix, 4 ℃, 3000rpm-5000rpm, 8min-10min is centrifugal, supernatant discarded, collect thalline, the sterilized water Eddy diffusion of mixed fermentation liquid cumulative volume 1/5-1/10 for thalline, adds mixed fermentation liquid 5%-15%(W/V) thalline protective material, on magnetic stirring apparatus, adsorb 30min-40min, or manually stir evenly standing 10-15min;
(4) compound nutrients and additive adds
In the bacterium liquid obtaining, add compound nutrients and additive after step (3) is collected and fixed, be settled to centrifugal front mixed fermentation liquid long-pending.
(5) filling
The liquid composite biological fertilizer of step (4) gained mycetome, nutrient substance, thalline protective material and additive is added in the Plastic Bottle of uv sterilisation, paste Product labelling, obtain liquid composite bio-fertilizer of the present invention.
Liquid composite biological fertilizer of the present invention can be used as biological pesticide, can directly apply to plant surface and spray, or be applied to seed soaking, filling root etc.
Application process: by liquid composite biological fertilizer dilution 200-300 of the present invention doubly, standing 30min, then stir evenly, directly spray in kind of plant blade face; Or after being diluted to 150-250 times, liquid composite biological fertilizer of the present invention is directly used in seed soaking; Also after liquid composite biological fertilizer of the present invention can being diluted to 100-200 times, for filling with root, every mu of ground applies composite biological fertilizer 200mL-400mL.
Beneficial effect:
(1) the present invention adopts bacillus thuringiensis, bacillus megaterium, colloid bacillus cereus, bacillus amyloliquefaciens as the production bacterial strain of composite microbiological fertilizer, is equipped with the production technique of optimization, makes contained each function yeast viable count in finished product all reach 10
8individual/more than mL, be 10-15 times that common commercially available prod bacterium is dense, there is high efficiency; That bacterial strain can reach is pest-resistant, phosphorus decomposing potassium, decompose organic, regulate the comprehensive functions such as soil pH.
(2) bacterial manure of the present invention is simultaneously composite with organic and inorganic complex function nutrient substance, for increasing soil fertility, improves crop growth environment, improvement soil and Promoting plant growth and has played active effect; Composite nutritive salt causes height to ooze system, and adds the additives such as organic acid and food preservatives, and safety non-toxic reduces system pH environment, strengthens the dormant state of microorganism, jointly reaches and suppresses the vital movement of thalline, the effect extending the shelf life; Liquid composite biological fertilizer of the present invention can be under the condition of room temperature lucifuge preservation at least 6 months.
(2) in the present invention, bacterial manure is composite with organic and inorganic complex function nutrient substance, adds the trace element of chelating state, plays between element the effect of mutually coordinating, relying on and restrict.Singularity due to the chelating salt structure adding, make electric charge in composite ion be tending towards neutral, in soil, avoid cationic exchange and can arrive fast crop root surface, more easily by Root Absorption, spray and on blade face, there is good mobility, easily by pore, absorbed, the albumen that can participate in body in without photosynthetic situation directly is synthetic, and its fertilizer efficiency is greater than the inorganic nitrogenous fertilizer of independent interpolation inorganic salt or equivalent.It is reported, the fertilizer efficiency of the micro-fertilizer of chelating state is 2～5 times of inorganic micro-fertilizer.
(3) bacterial manure of the present invention adds organic acid, and safety non-toxic reduces system pH environment, strengthens the dormant state of microorganism, jointly reaches and suppresses the vital movement of thalline, the effect extending the shelf life; Liquid composite biological fertilizer of the present invention can be under the condition of room temperature lucifuge preservation at least 6 months.
(4) the present invention adopts cheap raw material as the substratum of thalline liquid state fermentation, adopts efficient fermentation process, has reduced production cost, and a kind of method of preparing bio-bacterial manure of efficient cheapness is provided; The substratum of optimization of the present invention has the effect of fast fast-growing spore, makes thalline just can reach 90% gemma production rate at cultivation 12-15h.
(4) composite biological fertilizer provided by the present invention, in the planting experiment of the plants such as bracketplant, Plantula Brassicae chinensis, caraway, find, fertilising group is all obviously better than blank group at aspects such as leaf length, root length, plant height, fresh weight, output, simultaneously can pest-resistant evil, regulate the pH of alkaline soil;
(5) the contained four kinds of production bacterial strains of bacterial manure of the present invention are gemma bacterioid, growth and breeding ability is vigorous, strong adaptability to environment, is easy to grow soil is decided at the higher level but not officially announced, and when the zymotechnique that the present invention adopts and substratum make microorganism fermentation reach terminal, spore forming rate all reaches 90%-95%.
(6) the present invention has added thalline protective material as diatomite, kaolin etc., make thalline between the shelf lives in this material in dormant state, Cell protection tolerates the high osmotic pressure of bacterial manure system better.When it is used, thalline protective material can be dissolved very soon, and thalline Fast Stripping is rejuvenated, and just can make thalline depart from stripping unlike compact structure material demand long periods such as rhombspar or calcium carbonate.
(7) in the seaweed powder that the present invention adds, contain the organic substances such as indolylacetic acid, plant-growth kinetin, marine alga phenol, coordinate after function yeast and nutrient substance for plant husbandry can play more excellent drought resisting, anti-saline and alkaline infiltration, resist cold, sterilization and growth promoting function.
Embodiment
Below by specific embodiment narration the present invention.Unless stated otherwise, in the present invention, technique means used is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, but not limits the scope of the invention, and the spirit and scope of the invention are only limited by claims.To those skilled in the art, do not deviating under the prerequisite of essence of the present invention and scope various changes that the material component in these embodiments and consumption are carried out or change and also belong to protection scope of the present invention.
Embodiment 1
(1) preparation of substratum
No. 1 substratum: by percentage to the quality, 0.8%-1.2% peptone, 0.5%-1% yeast powder, 0.3%-1.0%NaCl, all the other are water, pH7.2-7.4.
No. 2 substratum: by percentage to the quality, 1%-2% N.F,USP MANNITOL or sucrose, 0.1 ‰-0.12 ‰ CaSO
42H
2o, 0.18 ‰-0.25 ‰ K
2hPO
4, 0.5%-0.6%CaCO
3, 0.15 ‰-0.2 ‰ MgSO
47H
2o, 0.2 ‰-0.25 ‰ NaCl, 1.4%-2% agar, all the other are water.
No. 3 substratum: by percentage to the quality, 1.0%-1.2% glucose, 0.5%-0.6%K
2hPO
43H
2o, 0.3%-0.5%KH
2pO
4, 0.1%-0.2% urea, 0.4%-0.6% yeast powder, 0.35%-0.45 ‰ MgSO
47H
2o, all the other are water, pH7.0-7.2.
No. 4 substratum: by percentage to the quality, 1% sucrose, 0.5%K
2hPO
43H
2o, 0.5 ‰ yeast powders, 0.02 ‰ FeCl
36H
2o, 0.4 ‰ CaCO
3, 0.2 ‰ MgSO
47H
2o, all the other are water, pH7.0.
(2) slant activation
Bacillus thuringiensis, bacillus megaterium, bacillus amyloliquefaciens are inoculated in to plate culture medium activation culture No. 1, by colloid bacillus cereus streak inoculation in No. 2 substratum activation culture.
(3) seed liquor preparation
The bacterium colony of picking bacillus thuringiensis, bacillus megaterium, bacillus amyloliquefaciens activation, is inoculated in respectively substratum No. 3, preparation seed liquor; The bacterium colony of picking colloid bacillus cereus activation, is inoculated in substratum No. 4, preparation seed liquor.
(4) fermentation culture
The seed liquor of cultured four bacterium is seeded to 5L fermentor tank separately, liquid amount 3L with 10% inoculum size respectively.Bacillus thuringiensis, bacillus megaterium, bacillus amyloliquefaciens, No. 3 substratum, 40 ℃, rotating speed 300rpm, ventilating ratio 1:1; Colloid bacillus cereus, No. 4 substratum, 40 ℃, rotating speed 400rpm, ventilating ratio 1:1; All ferment after 15-18h and collect fermented liquid.
After fermentation ends, after testing, each fermented liquid bacteria concentration all reaches 10
8individual/more than mL.
(5) four kinds of fermented liquids step (4) being collected mix, 4 ℃, 5000rpm, 8min is centrifugal, and supernatant discarded is collected thalline, total fermented liquid 1/10(V/V for thalline) sterilized water Eddy diffusion, add total fermented liquid 5%(W/V) talcum powder, on magnetic stirring apparatus, adsorb 40min, standing 15min;
(6) compound nutrients and additive adds
(5) collect and the bacterium liquid that fixedly obtains in add compound nutrients and additive, the mixed fermentation liquid being settled to before centrifugal is long-pending.
The parts by weight of compound nutrients consist of: potassium primary phosphate 100, EDTA-Ca25, EDTA-Mg80, EDTA-Zn0.18, EDTA-Mn0.3, EDTA-Fe0.1, EDTA-Cu0.08, boric acid 0.3, humic acids 15, soybean protein 100, aminoacids complex 50, seaweed powder 20.
The parts by weight of aminoacids complex consist of: L-Ala 6.5, arginine 6.8, aspartic acid 6.8, Gelucystine 2.5, glycine 3.2, Histidine 2.2, Methionin 6.5, methionine(Met) 3.2, phenylalanine 5.5, proline-4, Serine 8.5, Threonine 4.8, tryptophane 1.8, trorsine 14 .5, α-amino-isovaleric acid 7.5.
Described additive is oxysuccinic acid.
(8) filling
The above-mentioned liquid that mixes is added in the Plastic Bottle of uv sterilisation, and aluminium foil sealing is added a cover plastic cover again, pastes Product labelling, obtains liquid composite microbiological fertilizer of the present invention.
Described in the present embodiment, the composition of liquid composite biological fertilizer comprises; Parts by weight consist of, microbial cells 2, and compound nutrients 80, thalline protective material 20, additive 0.5, wherein microbial cells is the precipitation weight in wet base after centrifugal in fermented liquid.
Embodiment 2
(1) preparation of substratum
No. 1 substratum: by percentage to the quality, 1% peptone, 0.5% yeast powder, 0.3%NaCl, all the other are water, pH7.2;
No. 2 substratum: by percentage to the quality, 1% N.F,USP MANNITOL, 0.1 ‰ CaSO
42H
2o, 0.2 ‰ K
2hPO
4, 0.5%CaCO
3, 0.2 ‰ MgSO
47H
2o, 0.2 ‰ NaCl, 1.4% agar, all the other are water;
No. 3 substratum: by percentage to the quality, 1.0% glucose, 0.6%K
2hPO
4, 0.3%KH
2pO
4, 0.15% urea, 0.5% yeast powder, 0.4 ‰ MgSO
47H
2o, all the other are water, pH7.2
No. 4 substratum: by percentage to the quality, 1% sucrose, 0.5%K
2hPO
43H
2o, 0.5 ‰ yeast powders, 0.02 ‰ FeCl
36H
2o, 0.4 ‰ CaCO
3, 0.25 ‰ MgSO
47H
2o, all the other are water, pH7.0,
(2) slant activation
By each bacteria strain activation of 4 ℃ of preservations, after by bacillus thuringiensis, bacillus megaterium, bacillus amyloliquefaciens respectively streak inoculation on No. 1 plate culture medium, cultivate 1d for 30 ℃-37 ℃; Colloid bacillus cereus streak inoculation, in No. 2 substratum, is cultivated to 1-2d for 25 ℃-30 ℃.
(3) seed liquor preparation
The bacterium colony of picking bacillus thuringiensis, bacillus megaterium, bacillus amyloliquefaciens activation, is inoculated in respectively substratum No. 3, and 37 ℃, 200rpm, cultivates 16h;
The bacterium colony of picking colloid bacillus cereus activation, is inoculated in substratum No. 4, and 37 ℃, 200rpm, cultivates 16h; Now the dense bacterium of the bacterium of four bacterium all reaches 10
8individual/mL.
(4) fermentation culture
The seed liquor of cultured four bacterium is seeded to 5L fermentor tank separately, liquid amount 3L with 10% inoculum size respectively.Bacillus thuringiensis, bacillus megaterium, bacillus amyloliquefaciens, No. 3 substratum, 37 ℃, rotating speed 300rpm, ventilating ratio 1:1; Colloid bacillus cereus, No. 4 substratum, 37 ℃, rotating speed 400rpm, ventilating ratio 1:1; All ferment after 15-18h and collect fermented liquid.
After fermentation ends, after testing, each fermented liquid bacteria concentration is: bacillus megaterium 8 * 10
8individual/mL, bacillus thuringiensis 1 * 10
9individual/mL, bacillus amyloliquefaciens 5 * 10
8individual/mL, colloid bacillus cereus 6 * 10
8individual/mL, each bacterium brood cell production rate all reaches 95%.
(5) microorganism collection is with fixing
Four kinds of fermented liquids that step (3) is collected mix with volume ratio 3:2:3:4,4 ℃, 4000rpm, 10min is centrifugal, and supernatant discarded is collected thalline, total fermented liquid 1/5(V/V for thalline) sterilized water Eddy diffusion, add total fermented liquid 10%(W/V) diatomite, on magnetic stirring apparatus, adsorb 30min, standing 10min;
(6) compound nutrients and additive adds
(5) collect and the bacterium liquid that fixedly obtains in add compound nutrients and additive, the mixed fermentation liquid being settled to before centrifugal is long-pending.
The parts by weight of described compound nutrients consist of: potassium primary phosphate 150, EDTA-Ca28, MgSO
47H
2o+EDTA-Mg60, EDTA-Zn+ZnSO
47H
2o0.16, EDTA-Mn+MnSO
4h
2o0.4, EDTA-Fe0.2, EDTA-Cu0.07, boric acid 0.13, humic acids 10, soybean protein 40, aminoacids complex 50, seaweed powder 50.
Consisting of of aminoacids complex wherein: mass fraction is, L-Ala 6.1, arginine 6.6, aspartic acid 7.0, Gelucystine 2.3, glycine 3.5, Histidine 2.4, Methionin 6.4, methionine(Met) 3.1, phenylalanine 5.0, proline-4 .2, Serine 8.4, Threonine 5.0, tryptophane 1.6, trorsine 14 .3, α-amino-isovaleric acid 7.4.
Described additive is oxysuccinic acid and citric acid 1:2 volume mixture.
(8) filling
The above-mentioned liquid that mixes is added in the Plastic Bottle of uv sterilisation, and aluminium foil sealing is added a cover plastic cover again, pastes Product labelling, obtains liquid composite microbiological fertilizer of the present invention.
Described in the present embodiment, the composition of liquid composite biological fertilizer comprises; Parts by weight consist of, microbial cells 5, and compound nutrients 110, thalline protective material 30, additive 3.0, wherein microbial cells is the precipitation weight in wet base after centrifugal in fermented liquid.
The total viable count of liquid composite biological fertilizer of the present invention is 6.9 * 10
8individual/mL, pH4.0, ionic strength 80ms/cm.
The liquid composite biological fertilizer of preparing gained is standing in the room temperature higher than 30 ℃ not, lucifuge preservation, the total viable count of results of regular determination product, the total viable count of product did not have obvious decline within 3 months, in 6 months, do not decline to a great extent, during preservation to 12 month, the compound national standard that remains unchanged is desired is not less than 0.5 * 10
8individual/mL.
Total viable count of the liquid composite biological fertilizer different storage time of table 1
Above-mentioned bacterial classification is bought certainly: bacillus thuringiensis (Bacillus thuringiensis): microbial strains preservation center, Guangdong Province (5th floor, No. 100, Xianlie Middle Road, Guangzhou City province Institute of Micro-biology laboratory building, postcode 510070), numbering GIM1.151; Bacillus megaterium (Bacillus megatherium): Chinese common micro-organisms culture presevation administrative center (No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, postcode 100101), numbering CGMCC1.223; Bacillus amyloliquefaciens: Chinese industrial microbial strains preservation administrative center (No. 6 building of No. 24, Road, Jiuxianqiao, Chaoyang District, Beijing City institute, postcode 100015), numbering CICC23845; Colloid bacillus cereus (Bacillus mucilaginosu): Chinese industrial microbial strains preservation administrative center, numbering CICC23575.
Embodiment 3
According to the fermented liquid of four kinds of bacterium described in the described method Preparation Example 2 in (1) in embodiment 2-(4), four kinds of fermented liquid equal-volumes collecting are mixed, 4 ℃, 3000rpm, 8min is centrifugal, supernatant discarded, collect thalline, total fermented liquid 1/8(V/V for thalline) sterilized water Eddy diffusion, add total fermented liquid 15%(m/V) kaolin, on magnetic stirring apparatus, adsorb 30min, standing 10min; Add compound nutrients and additive, be settled to centrifugal front mixed fermentation liquid long-pending.Press the filling preservation of (8) described method in embodiment 1, meanwhile, with do not add thalline protective material with the product of additive for contrasting.Its total viable count of results of regular determination.
The parts by weight of described compound nutrients consist of: potassium primary phosphate 50, EDTA-Ca30, MgSO
47H
2o+EDTA-Mg50, EDTA-Zn0.15, EDTA-Mn+MnSO
4h
2o0.5, EDTA-Fe0.3, EDTA-Cu0.06, boric acid 0.2, humic acids 5, soybean protein 20, aminoacids complex 50, seaweed powder 100.
Wherein aminoacids complex forms with embodiment 2.
Described additive consists of lactic acid and citric acid 1:1 volume mixture.
Described in the present embodiment, the composition of liquid composite biological fertilizer comprises; Parts by weight consist of, microbial cells 3, and compound nutrients 130, thalline protective material 20, additive 1.5, wherein microbial cells is the precipitation weight in wet base after centrifugal in fermented liquid.
The liquid bacterial manure of table 2 is at total viable count (individual/mL) of different times
The present invention has added after thalline protective material and additive, keep bacterium number significantly to reduce, and thalline is energetic in 6 months, meets total viable count in 3 months that GB NY/T798-2004 require and is not less than 0.5 hundred million/mL.
Embodiment 4
According to method described in embodiment 2, prepare liquid composite biological fertilizer, by 200 times of this bacterial manure dilutions, the solution 200mL getting after dilution applies in the flowerpot (punching at the bottom of basin) that 300g soil is housed, is loaded on 12 * 10cm, after solution soaks into soil, by the suitable bracketplant plant young (unrooted of growing way, 4 leaves, the long 6cm of leaf, average plant height 6cm, fresh weight 1g) insert in soil and cultivate, water every day, and every 7d topdresses once, blank group is set simultaneously, and when experimental group is applied fertilizer, control group applies equivalent tap water.Two groups of plant growing ways of routine observation, measure soil pH, during 45d, observe plant, measure the parameters such as plant height, leaf length, root length, fresh weight, all than blank, demonstrate clear superiority: the well developed root system of fertilising plant, Ye Changyou rises appreciably, and newborn blade is also more than blank, and fresh weight gaining rate reaches 400%.Meanwhile, after fertilising, the soil is porous, and soil pH has obvious reduction than unmanured soil pH, and this is that this pH to the saline-alkali soil of north of China has good regulating effect because thalline has been secreted acid to soil in process of growth.
Plant parameter during table 3 bracketplant plantation 45d
Table 4 fertilising soil pH and the soil pH comparison of not applying fertilizer
Embodiment 4
According to method described in embodiment 2, prepare liquid composite biological fertilizer, by 250 times of product dilutions, spray in the blade surface of Chinese cabbage, Plantula Brassicae chinensis, wild cabbage, broccoli, every 7 days, spray once, can obviously reduce lepidoptera pest harm, as small cabbage moth.
Embodiment 5
According to method described in embodiment 3, prepare liquid composite biological fertilizer, product is imposed on to the crops such as tomato, capsicum, eggplant, cucumber, dilute 150 times and fill with root, every mu of former solution 200mL, after this every 7d, impose once, dilute 250 times, after finishing in crop cycle, each crop yield all can improve more than 30%.
Claims (8)
1. a liquid composite biological fertilizer, the composition of described liquid composite biological fertilizer comprises microbial cells, compound nutrients, thalline protective material and additive; Its parts by weight consist of, microbial cells 2-5, and compound nutrients 80-130, thalline protective material 10-30, additive 0.5-3.0, wherein microbial cells is the precipitation weight in wet base after centrifugal in fermented liquid;
The composition of described microbial cells comprises: bacillus thuringiensis (Bacillus thuringiensis), bacillus megaterium (Bacillusmegaterium), colloid bacillus cereus (Bacillus mucilaginosus), bacillus amyloliquefaciens (Bacillus amylolquefaciens), wherein total viable count reaches 5 * 10
8-5 * 10
9individual/mL;
The parts by weight of described compound nutrients consist of: potassium primary phosphate 50-150, EDTA-Ca25-30, EDTA-Mg or EDTA-Mg and MgSO
47H
2the two mixture 50-80 of O, EDTA-Zn or EDTA-Zn and ZnSO
47H
2o or the two mixture 0.15-0.18, EDTA-Mn or EDTA-Mn and MnSO
4h
2o or the two mixture 0.3-0.5, EDTA-Fe0.1-0.3, EDTA-Cu0.06-0.08, boric acid 0.1-0.3, humic acids 5-15, soybean protein 20-100, aminoacids complex 50, seaweed powder 20-100;
Described thalline protective material is one or more in cell immobilization carrier;
Described additive is one or more in organic acid.
2. liquid composite biological fertilizer according to claim 1, is characterized in that, the parts by weight of described aminoacids complex consist of: L-Ala 6.0-6.5, arginine 6.5-6.8, aspartic acid 6.8-7.2, Gelucystine 2.2-2.5, glycine 3.2-3.5, Histidine 2.2-2.5, Methionin 6.4-6.5, methionine(Met) 3.0-3.2, phenylalanine 5-5.5, proline-4-4.2, Serine 8.4-8.5, Threonine 4.8-5.0, tryptophane 1.5-1.8, trorsine 14 .2-4.5, α-amino-isovaleric acid 7.4-7.5.
3. liquid composite biological fertilizer according to claim 1, is characterized in that, described thalline protective material is the mixture of a kind of in diatomite, kaolin, talcum powder or at least two kinds.
4. liquid composite biological fertilizer according to claim 1, is characterized in that, described additive is the mixture of a kind of in oxysuccinic acid, lactic acid, citric acid or at least two kinds.
5. liquid composite biological fertilizer according to claim 1, it is characterized in that, in described microbial cells, four kinds of bacterium viable count ratios are as follows: bacillus thuringiensis: bacillus megaterium: colloid bacillus cereus: bacillus amyloliquefaciens=1-2:2-3:2-3:1-2.
6. according to the production method of the arbitrary described liquid composite biological fertilizer of claim 1-5, comprise the steps:
(1) actication of culture and seed liquor preparation
Bacillus thuringiensis, bacillus megaterium, colloid bacillus cereus, Bacillus amyloliquefaciens strain are activated respectively and cultivated and make seed liquor;
(2) fermentation culture
The seed liquor of cultured bacillus thuringiensis, bacillus megaterium, bacillus amyloliquefaciens is seeded to 5L fermentor tank with the inoculum size of 8%-10% respectively, fermention medium is substratum No. 3, and 37 ℃～40 ℃, rotating speed 300rpm, ventilating ratio 1:1, collects fermented liquid after fermentation 15-18h;
The seed liquor of cultured colloid bacillus cereus is seeded to 5L fermentor tank with the inoculum size of 8%-10%, and fermention medium is No. 4 substratum, 37 ℃～40 ℃, and rotating speed 400rpm, ventilating ratio 1:1, collects fermented liquid after fermentation 15-18h;
Described No. 3 substratum form: by percentage to the quality, and 1.0%-1.2% glucose, 0.5%-0.6%K
2hPO
43H
2o, 0.3%-0.5%KH
2pO
4, 0.1%-0.2% urea, 0.4%-0.6% yeast powder, 0.35%-0.45 ‰ MgSO
47H
2o, all the other are water, pH7.0-7.2;
Described No. 4 substratum: by percentage to the quality, 1% sucrose, 0.5%K
2hPO
43H
2o, 0.5 ‰ yeast powders, 0.02 ‰ FeCl
36H
2o, 0.4 ‰ CaCO
3, 0.2 ‰ MgSO
47H
2o, all the other are water, pH7.0;
(3) microorganism collection is with fixing
Four kinds of fermented liquids that step (2) is collected mix, 4 ℃, 3000rpm-5000rpm, 8min-10min is centrifugal, supernatant discarded, collect thalline, the sterilized water Eddy diffusion of mixed fermentation liquid cumulative volume 1/5-1/10 for thalline, adds mixed fermentation liquid 5%-15%(W/V) thalline protective material, on magnetic stirring apparatus, adsorb 30min-40min, or manually stir evenly standing 10-15min;
(4) compound nutrients and additive adds
In the bacterium liquid obtaining in step (3), add nutrient substance and additive, be settled to centrifugal front mixed fermentation liquid long-pending;
(5) filling
The liquid composite biological fertilizer of step (4) gained mycetome, nutrient substance, thalline protective material and additive is added in the Plastic Bottle of uv sterilisation, paste Product labelling, obtain described liquid composite bio-fertilizer.
7. the production method of liquid composite biological fertilizer according to claim 6, is characterized in that, comprises the steps:
(1) actication of culture and seed liquor preparation
Bacillus thuringiensis, bacillus megaterium, colloid bacillus cereus, Bacillus amyloliquefaciens strain are activated respectively and cultivated and make seed liquor;
(2) fermentation culture
The seed liquor of cultured four bacterium is seeded to 5L fermentor tank separately, liquid amount 3L with 10% inoculum size respectively.Bacillus thuringiensis, bacillus megaterium, bacillus amyloliquefaciens, No. 3 substratum, 37 ℃, rotating speed 300rpm, ventilating ratio 1:1; Colloid bacillus cereus, No. 4 substratum, 37 ℃, rotating speed 400rpm, ventilating ratio 1:1; All ferment after 15-18h and collect fermented liquid;
No. 3 substratum: by percentage to the quality, 1.0% glucose, 0.6%K
2hPO
4, 0.3%KH
2pO
4, 0.15% urea, 0.5% yeast powder, 0.4 ‰ MgSO
47H
2o, all the other are water, pH7.2;
No. 4 substratum: by percentage to the quality, 1% sucrose, 0.5%K
2hPO
43H
2o, 0.5 ‰ yeast powders, 0.02 ‰ FeCl
36H
2o, 0.4 ‰ CaCO
3, 0.25 ‰ MgSO
47H
2o, all the other are water, pH7.0;
After fermentation ends, after testing, each fermented liquid bacteria concentration is: bacillus megaterium 8 * 10
8individual/mL, bacillus thuringiensis 1 * 10
9individual/mL, bacillus amyloliquefaciens 5 * 10
8individual/mL, colloid bacillus cereus 6 * 10
8individual/mL, each bacterium brood cell production rate all reaches 95%.
(3) microorganism collection is with fixing
Four kinds of fermented liquids that step (2) is collected mix with volume ratio 3:2:3:4,4 ℃, 4000rpm, 10min is centrifugal, and supernatant discarded is collected thalline, total fermented liquid 1/5(V/V for thalline) sterilized water Eddy diffusion, add total fermented liquid 10%(W/V) diatomite, on magnetic stirring apparatus, adsorb 30min, standing 10min;
(4) compound nutrients and additive adds
(3) collect and the bacterium liquid that fixedly obtains in add compound nutrients and additive, the mixed fermentation liquid being settled to before centrifugal is long-pending;
The parts by weight of described compound nutrients consist of: potassium primary phosphate 150, EDTA-Ca28, MgSO
47H
2o+EDTA-Mg60, EDTA-Zn+ZnSO
47H
2o0.16, EDTA-Mn+MnSO
4h
2o0.4, EDTA-Fe0.2, EDTA-Cu0.07, boric acid 0.13, humic acids 10, soybean protein 40, aminoacids complex 50, seaweed powder 50;
Consisting of of aminoacids complex wherein: mass fraction is, L-Ala 6.1, arginine 6.6, aspartic acid 7.0, Gelucystine 2.3, glycine 3.5, Histidine 2.4, Methionin 6.4, methionine(Met) 3.1, phenylalanine 5.0, proline-4 .2, Serine 8.4, Threonine 5.0, tryptophane 1.6, trorsine 14 .3, α-amino-isovaleric acid 7.4;
Described additive is oxysuccinic acid and citric acid 1:2 volume mixture;
(8) filling
The above-mentioned liquid that mixes is added in the Plastic Bottle of uv sterilisation, and aluminium foil sealing is added a cover plastic cover again, pastes Product labelling, obtains described liquid composite microbiological fertilizer;
The parts by weight of described liquid composite biological fertilizer consist of, microbial cells 5, and compound nutrients 110, thalline protective material 30, additive 3.0, wherein microbial cells is the precipitation weight in wet base after centrifugal in fermented liquid.
8. the arbitrary described liquid composite biological fertilizer of claim 1-5 is as the purposes of biological pesticide.
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