CN103739315A - Preparation method of green microecological compound fertilizer - Google Patents
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Abstract
The invention discloses a preparation method of a green microecological compound fertilizer, belonging to the technical field of agricultural fertilizers. The preparation method of the green microecological compound fertilizer comprises the following step: proportionally combining an Aspergillus niger nutrient material, a Bacillus mucilaginosus microbial inoculum, a phosphate-solubilizing Bacillus megaterium microbial inoculum, a Bacillus subtilis nutrient material, an Aspergillus awamori nutrient material and a lactobacillus plantarum microbial inoculum to obtain the microecological compound fertilizer. The fertilizer is prepared by fermenting compound microbes, can improve the microecological environment of soil, and provides beneficial conditions for crop growth. The fertilizer can improve the soil. The beneficial microbes in the fertilizer can generate saccharides (accounting for 0.1% of soil organic matters), the saccharides can be combined with the plant mucilage, mineral embryo and organic colloid to improve the crumb structure of the soil, enhance the physical properties of the soil and reduce the loss of the soil particles; and the fertilizer can participate in humification and improve the physical behaviors of the soil under certain conditions, thereby being beneficial to increasing the soil fertility.
Description
Technical field
The present invention relates to the preparation method of the micro-ecological composite fertilizer of a kind of green, belong to agricultural fertilizer technical field.
Background technology
Microorganism fertilizer is a kind ofly with microbial life activity and product thereof, to cause farm crop to obtain the microorganism live body goods of specific fertilizer effect, it at culture fertility, improve chemical fertilizer utilization ratio, suppress farm crop there is to irreplaceable effect the aspects such as utilizations of becoming thoroughly decomposed of the absorption of the objectionable impuritiess such as heavy metal and agricultural chemicals, purification and rehabilitating soil, promotion agricultural crop straw and municipal wastes, raising quality of agricultural product.The efficacy exertion of microbial fertilizer is mainly by the synergism to traditional fertilizer, fertilizer, improvement activation to soil, and the modes such as physiological action of microorganism realize.Therefore, microbial fertilizer has just had multiple fertility and the effect of chemical fertilizer, fertilizer, beneficial organism bacterium, is activated fertilizer nutrient, improves effect of fertilizer, Crop Improvement quality, promotes the desirable fertilizer of agricultural produce synergy.Microbial fertilizer is live body fertilizer, and a large amount of beneficial microorganism vital movements that its effect mainly contains by it complete.Only have these beneficial microorganism vital movements of working as to complete.Only have when these beneficial microorganisms are in vigorous breeding and metabolic situation, Substance Transformation and useful meta-bolites could constantly form.Therefore, beneficial microorganism kind in microbial fertilizer, whether vital movement is vigorous is the basis of its validity, and is to take the form of the principal elements such as nitrogen, phosphorus, potassium and how many for basic unlike other fertilizer.Just because of microbial fertilizer is the preparation of living, so its fertilizer efficiency and number of viable, intensity and ambient environmental conditions are closely related, comprise temperature, moisture, potential of hydrogen, nutritional condition and formerly live in indigenous microorganism repulsive interaction in soil and have certain influence.
Because China lack to drop in research aspect microbial fertilizer for a long time, make that the microbial fertilizer industry of China still exists that integral level is not high, technological innovation is not enough, quality product and effect show understable problem.At agricultural sustainable development, become today of human consensus, these problems have become microorganism fertilizer industry letter " bottleneck " to be got through.
Summary of the invention
Technical problem to be solved by this invention is to overcome the deficiencies in the prior art, and the preparation method of the micro-ecological composite fertilizer of a kind of green is provided: be combined as in proportion micro-ecological composite fertilizer
A preparation method for the micro-ecological composite fertilizer of green, comprises the steps: aspergillus niger culture, colloid bud pole bacteria agent, phosphorus decomposing Bacillus megaterium microbial inoculum, subtilis culture, Aspergillus awamori culture, plant lactobacillus agent to be combined as in proportion micro-ecological composite fertilizer;
The micro-ecological composite fertilizer of described green parts by weight consist of: aspergillus niger culture 5-10 part, colloid bud pole bacteria agent 3-8 part, phosphorus decomposing Bacillus megaterium microbial inoculum 3-8 part, subtilis culture 5-12, Aspergillus awamori culture 4-7, plant lactobacillus agent 2-4 part.
Aspergillus niger (Aspergillus niger) Li-2013-03 preserving number is CGMCC NO.7927.
Described subtilis culture preparation: cultivate subtilis from inclined-plane switching, the seed liquor after spreading cultivation is step by step transferred in fermentor tank, the complete fermented liquid that ferments obtains subtilis culture through Plate Filtration, after dry.
The preparation method of plant lactobacillus agent: from inclined-plane switching culturing plants Bacterium lacticum, the seed liquor after spreading cultivation is step by step transferred in fermentor tank, and the complete fermented liquid that ferments is concentrated in vacuo to 45% of original volume through low-temperature negative-pressure, obtains bacterium concentrated solution.Add carrier: in concentrated solution, add the carrier mixing, mix; The weight ratio of concentrated solution and carrier is 0.5-0.6:1, and vehicle group becomes: CaCO
320-30 part, dextrin 10-15 part.Fluidised bed drying, 50 ℃ of drying temperatures.
Aspergillus awamori culture preparation: spawn culture, solid fermentation is cultivated: spore liquid is inoculated in aspergillus oryzae solid state fermentation culture material, and 26-33 ℃ is cultured to mycelia and covers with culture material, and low temperature fluidized-bed dry, pulverize dry thing.
Aspergillus niger culture preparation: spawn culture, adopt common solid fermentation cultural method preparation: spore liquid is inoculated in solid state fermentation culture material, 26-33 ℃ is cultured to mycelia and covers with culture material, dry, pulverize dry thing.
Rui Gu bio tech ltd, Baoding provides colloid bacillus cereus bacterium powder;
Beneficial effect:
Fertilizer of the present invention can strengthen the drought-resistant ability of crop.Colloid bud pole bacterium in composite bacteria of the present invention can secrete the growth-promoting substances such as growth hormone material and Plant hormones regulators,gibberellins material, extraneous root strong sprout; Decompose element silicon in soil and, for plant utilization, the wax layer of plant is thickened, improve plant water keeping ability; Can promote soil aggregate to form, keep soil from packing together; Spoiled soil capillarity, prevents soil water evaporation.
Fertilizer of the present invention is formed by compound microorganism ferments, can improve the micro-ecological environment of soil, for the growths of farm crop provides beneficial conditions.Fertilizer of the present invention can be improved soil.In fertilizer, beneficial microorganism can produce glucide, account for 0.1% of the soil organism, with plant mucilage, mineral idiosome and organic colloid combine, can improve soil aggregate, strengthen the physicals of soil and the loss of minimizing soil particle, under certain conditions, can also participate in soil ulmin and form.So use microbial fertilizer, can improve soil physical property, be conducive to increase soil fertility.
This product using method is simple, every mu of ground usage quantity 0.3-1 kilogram, and cost is only 80-100 unit/mu, spray on seed dressing or the front earth's surface of pouring water vegetative period.
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.
Aspergillus niger strain provided by the invention (Aspergillus niger) Li-2013-03 is the many wheels of aspergillus niger (Aspergillus niger) the Li-2010 warp nitrosoguanidine mutagenesis by a strain cellulase-producing product of laboratory preservation, then mutant strain step-sizing is eliminated, finally strain excellent is obtained producing Aspergillus niger strain (Aspergillus niger) Li-2013-03 of high activity cellulase through leavening property test screen.
The bacterial strain of the high activity cellulase of product provided by the invention is specially aspergillus niger (Aspergillus niger) Li-2013-03.This bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on July 15th, 2013 and (is called for short CGMCC, address is: No. 3, No. 1, North Star West Road, Chaoyang District, BeiJing, China city institute, postcode 100101), preserving number is CGMCC NO.7927.
The screening method that produces high activity cellulase bacterial strain, comprises the following steps:
1) slant culture: by original Aspergillus niger strain (Aspergillus niger) Li-2010 streak inoculation slant medium, cultivate 2～3d for 30 ℃, until mycelium ripe, produce a large amount of black spores.Described slant medium is composed as follows: 12 ° of Brix wort l000mL, pH value nature, 121 ℃ of sterilizing 20min;
2) spore suspension preparation (following steps all operate under aseptic condition): add 15mL sterilized water to test tube slant, spore is scraped, with filter paper filtering, pour filtered solution into sterilizing and be added with in the 150mL triangular flask of 5-10 grain sterile glass beads, triangular flask is put into shaking table vibration 10-15min, spore is disperseed.
3) nitrosoguanidine (NTG) mutagenesis
A. with sterilized water, spore suspension is adjusted to and is diluted to 10
6-10
7individual/mL.
B. get 10mL bacteria suspension and be transferred in 100mL triangular flask, add the NTG of 10mg, being mixed with final concentration is the NTG solution of 10mg/mL, and adds 4-5 to drip acetone, is beneficial to NTG and dissolves.
C. 200rpm oscillatory reaction 30min at 30 ℃, the centrifugal 10min of 5000rpm collects thalline, with stroke-physiological saline solution washing for several times, stopped reaction.
D. suitably dilution is adjusted to 10 by spore concentration
3individual/mL, gets last dilution bacterium liquid 0.2mL, and dilution spread is on Mierocrystalline cellulose-Congo red plate screening substratum.200 of 30 ℃ of cultivation bacterial strains that after 2～3 days, picking transparent circle/colony diameter is larger.(described Mierocrystalline cellulose-Congo red plate screening substratum is composed as follows: cellulose powder 10g, Congo red 0.2g, ammonium sulfate 5g, magnesium sulfate 0.25g, potassium primary phosphate 1g, sodium-chlor 0.1g, gelatin 2g, agar 20g, tap water constant volume 1000mL, pH value 5-6,121 ℃ of sterilizing 20min).
E. sieve again: the 200 strain bacterium that obtain are inoculated in to slant medium with aseptic toothpick respectively, and 30 ℃ are cultured to spore and are paved with inclined-plane.Respectively spore is fermented to be inoculated under aseptic washing to be equipped with in the 250mL triangular flask that 50mL sieves substratum again, inoculum size 10%(v/v), 30 ℃, 100r/min are cultivated 96h, measure respectively the cellulase activity of each bacterial strain.(described sieve again substratum composed as follows: cellulose powder 50g, ammonium sulfate 5g, magnesium sulfate 0.25g, potassium primary phosphate 1g, sodium-chlor 0.1g, tap water constant volume 1000mL, pH value 5-6,121 ℃ of sterilizing 20min).Choose the bacterial strain that cellulose enzyme vigor is the highest and carry out amplification test.
4) genetic stability test
Li-2013-03 bacterial strain is gone down to posterity for continuous ten times on inclined-plane, and the method for sieving again by shaking flask detects the fermentation situation after at every turn going down to posterity.Experiment discovery is gone down to posterity for continuous ten times on inclined-plane, and this bacterial classification proterties does not have considerable change, and property indices is all normal, illustrates that the genetic stability of this bacterial classification is stronger.
5) scale-up
1. seed culture: by the highest strains A spergillus nigerLi-2013-03 access 500mL triangular flask of cellulose enzyme vigor, 100 milliliters of seed culture medium loading amounts, 30 ℃, 150rpm shaking table are cultivated 72-96h.
3. seed tank culture: by seed liquor with 10%(v/v) inoculum size access is equipped with in the 10L fermentor tank of 7.5L fermented liquid, controlling pH value constant is 6.0 ± 0.2,30 ± 0.1 ℃ of culture temperature, stirring velocity 300rpm, ventilation (v/v) 1:0.8-1.2, incubation time 96h, dissolved oxygen 20-30%.Described fermention medium consists of: cellulose powder 100g, ammonium sulfate 5g, magnesium sulfate 0.25g, potassium primary phosphate 1g, sodium-chlor 0.1g, tap water constant volume 1000mL, pH value 5-6,121 ℃ of sterilizing 20min.
After fermentation ends, get fermented supernatant fluid (crude enzyme liquid) and carry out enzyme activity detection after measured, circumscribed beta-glucanase, Endo-β-glucanase, beta-glucosidase and the filter paper enzyme activity of strains A spergillus nigerLi-2013-03 reach respectively 620U/mL, 1289U/mL, 456U/mL and 732U/mL, have improved 9.21 times, 7.43 times, 8.15 times and 10.31 times respectively than starting strain Aspergillus niger Li-2010.
Phosphorus decomposing Bacillus megaterium is a kind in Bacillus (Bacillus).Motion, forms pod membrane, aerobic.From glucose, produce acid, also often from pectinose and N.F,USP MANNITOL, produce acid.Hydrolyzed starch, does not produce lecithinase, and VP is negative.The organophosphorus that can decompose in soil becomes the available rapid available phosphorus of plant.
The agent of phosphorus decomposing bacillus megaterium is provided by Cangzhou prosperous generation thing technical institute, address: liberation West Road, Canal Zone, Hebei China Cangzhou City chin or cheek and Building A, international business affairs center 1 district 807-812.
Rui Gu bio tech ltd, Baoding provides colloid bacillus cereus bacterium powder.
Subtilis provided by the invention (Bacillus subtilis) Li-2013-02.This bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on July 15th, 2013, and (be called for short CGMCC, address is: No. 3, No. 1, North Star West Road, Chaoyang District, BeiJing, China city institute), preserving number is CGMCC No.7926.
Described bacterial strain feature is as follows:
Described bacterial strain colony colour on solid plate is oyster white, and surface drying is opaque, and neat in edge, for having the aerobic bacteria of mobility.Microscopy is elongated rod shape, and gramstaining is positive.This bacterium can utilize Citrate trianion, and nitrate reductase, V-P test into positive.
Described subtilis (Bacillus subtilis) Li-2013-02 is preserved in the high temperature resistant α-amylase of product in this laboratory subtilis Li-2013 by a strain obtains through UV-LiCl-ethyl sulfate Mutation screening, specifically screens step as follows:
(1) preparation of bacteria suspension
By in the mono-bacterium colony of the Li-2013 growing after plate streaking separation access seed culture medium, 100r/min, cultivates after 12h for 40 ℃, uses physiological saline washed twice after getting 1mL medium centrifugal, and in resuspended and 9mL physiological saline.
(2) UV-LiCl-ethyl sulfate complex mutation
Bacteria suspension is placed in to aseptic flat board, is 30cm in distance, stirs and irradiate 100s under the ultraviolet lamp of power 15w.To after gradient dilution, coat lithium chloride flat board through the bacterium liquid irradiating, and contrast with the bacterium liquid dilution painting flat board without uv irradiating.Above-mentioned coating is dull and stereotyped uniformly, with cloth or the newspaper of black, wrap, put 40 ℃ and cultivate 48h, on the flat board of bacterium colony, filter out hydrolysis circle and choose to inclined-plane and preserve with colony diameter ratio the maximum growing, after purifying, be mixed with bacteria suspension, after gradient dilution, fully mix with ethyl sulfate stoste, and process 40min in 40 ℃ of concussions, the bacterium liquid of processing is coated to lithium chloride flat board after gradient dilution.
(3) primary dcreening operation of high-yield strains
Above-mentioned coating is dull and stereotyped uniformly, put 40 ℃ and cultivate 48h, go out hydrolysis circle and choose to inclined-plane and preserve with colony diameter ratio the greater growing on the flat board of bacterium colony primary dcreening operation, after purifying, obtain three strain bacterium Li-2013-01, Li-2013-02, Li-2013-03
(4) shake flask fermentation sieves again
By the three strain bacterium Li-2013-01 that obtain, Li-2013-02, Li-2013-03 carries out shake flask fermentation in the 250mL shaking flask that contains 30mL fermention medium, seed inoculum size 10% (V/V), 40 ℃, 100r/min are cultivated 72h, and centrifuging and taking fermented supernatant fluid makes crude enzyme liquid.
(5) enzyme activity determination
The definition of Mei Huo unit: 1mL crude enzyme liquid, under 105 ℃, pH4.2 condition, 1min liquefaction 1mg Zulkovsky starch, is 1 enzyme activity unit, represents with U/mL.
After measured, bacterial strain Li-2013-02, is stable superior strain, and enzyme work reaches 30000U/mL, than original strain enzyme, lives and improves 1.6 times.
Described lithium chloride is dull and stereotyped: starch 1%, and peptone 1%, (NH)
2sO
40.4%, K
2hPO
40.8%, CaCl
20.2%, lithium chloride 0.9%, agar 2%.
Described seed culture medium: yeast powder 0.5%, peptone 1%, Zulkovsky starch 1%, NaCl1%.
Described fermention medium: Semen Maydis powder 5%～15%, soybean cake powder 4%～10%, (NH) 2SO40.4%, K2HPO40.8%, CaCl20.2%.
Described shake-flask culture condition: this bacterium in the 250mL shaking flask that contains 30mL fermention medium, inoculum size 10% (V/V), 100r/min, 40 ℃ of fermentation culture 72h.
By bacterial strain Li-2013-02 fermentation, obtained a kind of high-temperature resistant alpha-amylase, its zymologic property is as follows:
(1) this enzyme thermal adaptation a wider range, optimum temperature is between 105-115 ℃, and better 110 ℃ of following temperature stabilities of preserving, and 115 ℃ above to preserve long-time temperature stabilities poor.
(2) this enzyme optimal reaction pH value is 4.2.Between pH value 3.0-7.0, all having high enzyme vigor, is 3.0 o'clock enzyme complete stabilities alive in pH value.
(3) enzymic activity: by mutant strain Li-2013-02 provided by the present invention, the high temperature resistant α-amylase enzyme activity of preparation is 30000-35000U/ml.
1, the present invention uses the method for UV-LiCl-ethyl sulfate complex mutation to obtain the subtilis Li-2013-02 of a strain high-yield thermostable α-amylase, and this bacterial strain has acidproof, thermotolerance by force, produces the high feature of enzyme activity.
2, there is the high temperature resistant α-amylase enzyme activity of this bacterial strain production gained up to 30000-35000u/ml; Applicable temperature scope is 25-115 ℃, and 110 ℃ of optimal reactive temperatures, at 110 ℃ of enzymes complete stability alive; Applicable pH value in reaction scope is 3.0-7.0, in pH value, be 3.0 o'clock enzyme complete stabilities alive, optimal reaction pH value is 4.2, higher than existing high temperature resistant α-amylase enzyme activity, enzyme effect optimum pH wide scope, heatproof degree is high, is particularly suitable for that temperature of reaction is high, liquefaction process and Mashing process the industrialization demand of depositing.
Embodiment 1
Provide a kind of green micro-ecological composite fertilizer: parts by weight consist of: 6 parts of aspergillus niger cultures, 5 parts of colloid bud pole bacteria agents, 5 parts of phosphorus decomposing Bacillus megaterium microbial inoculums, subtilis culture 10, Aspergillus awamori culture 6,3 parts of plant lactobacillus agent.
The bacterial classification adopting is as follows:
Subtilis (Bacillus subtilis subsp) CGMCC7926
Plant lactobacillus (Lactobacillus plantarum) CICC20764
Aspergillus awamori (Aspergillus awamori) CGMCC3.6484
Aspergillus niger (Aspergillus niger) Li-2013-03 preserving number is CGMCC NO.7927.
The preparation method of Aspergillus awamori microbial inoculum:
Technical scheme is as follows:
Slant strains activation culture: Aspergillus awamori slant strains is transferred on slant medium, cultivates 3 days for 27 ℃.
Solid first order seed is cultivated: picking Aspergillus awamori slant strains accesses in 500 milliliters of triangular flasks that 100 grams of substratum are housed carries out seed culture, cultivates 3 days for 30 ℃.
Solid secondary seed is cultivated: above-mentioned cultured solid first order seed is stirred as adding after fragment and in 5000 milliliters of triangular flasks that 1000 grams of substratum are housed, carries out seed culture, culture condition: cultivate 3 days for 30 ℃.
Solid fermentation is cultivated: secondary shake-flask seed is pulverized, added to be equipped with in the fermentation vat of sterilising medium or pallet to mix rear cultivation, bent material culture temperature is controlled at 26-35 ℃, humidity 80-90%, every 10 hours stirrings once, incubation time 5-7 days; The cultivation of solid koji material adopts conventional bent material culture technique; Treat that culture material covers with mycelia and can finish to cultivate, substratum is in advance through high temperature steaming sterilising treatment, and sterilising conditions is controlled 121 ℃ of temperature, 1 hour time.
Drying and crushing: fermentation ends culture material is dried on fluidized-bed or other drying plants, and drying temperature is controlled at 60 ℃, is dried to moisture content below 10%, then solid culture medium is pulverized, and crushing material aperture is more than 60 orders.
Substratum forms: solid material: wheat bran 80%, and soybean cake powder 10%, W-Gum 10%, adds equivalent tap water; Initial pH nature.
The preparation method of subtilis culture:
1. the acquisition of fermented liquid: adopt slant strains to spread cultivation step by step and obtain fermentation of bacillus subtilis liquid;
(1) first order seed is cultivated: subtilis slant strains is accessed in 500 ml shake flasks to 100 milliliters of substratum loading amounts, 180 revs/min of rotary shaking tables, 30 ℃ of culture temperature, incubation time 24 hours;
(2) secondary seed is cultivated: by first order seed, according in 500 milliliters of secondary seed shaking flasks of inoculum size access of 10%, culture condition is identical with first order seed;
(3) three grades of seed culture: secondary seed is accessed in 5000 milliliters of three grades of seed shaking flasks to 1000 milliliters of substratum loading amounts, 100 revs/min of rotary shaking tables, 30 ℃ of culture temperature, incubation time 24 hours with 10% inoculum size;
(4) first class seed pot is cultivated: three grades of seeds be take to the first class seed pot that 10% inoculum size access cubic capacity is 150L, fermention medium loading amount 100L, 28 ℃ of culture temperature, 100 revs/min of stirring velocitys, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa, incubation time 24 hours;
(5) fermentation culture: it is 1.5 tons of secondary seed tanks that first class seed pot bacterial classification be take to 10% inoculum size access cubic capacity, 1 ton of fermention medium loading amount, 28 ℃ of culture condition culture temperature, 100 revs/min of stirring velocitys, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa, incubation time 24 hours.
Substratum forms: glucose 6%, yeast extract 1%, peptone 0.2%, CaCO31%, pH6.8.
The preparation method of plant lactobacillus agent:
(1) first order seed is cultivated: plant lactobacillus bacterial classification is accessed in 500 ml shake flasks to 100 milliliters of substratum loading amounts, 30 ℃ of culture temperature, incubation time 24 hours;
(2) secondary seed is cultivated: by first order seed, according in 500 milliliters of secondary seed shaking flasks of inoculum size access of 10%, culture condition is identical with first order seed;
(3) three grades of seed culture: secondary seed is accessed in 5000 milliliters of three grades of seed shaking flasks to 1000 milliliters of substratum loading amounts, 30 ℃ of culture temperature, incubation time 24 hours with 10% inoculum size;
(4) first class seed pot is cultivated: three grades of seeds be take to the first class seed pot that 5% inoculum size access cubic capacity is 150L, fermention medium loading amount 100L, 30 ℃ of culture temperature, tank pressure 0.05Mpa, incubation time 18 hours;
(5) fermentor cultivation: it is 3 tons of secondary seed tanks that first class seed pot bacterial classification be take to 5% inoculum size access cubic capacity, 2 tons of fermention medium loading amounts, 30 ℃ of culture condition culture temperature, tank pressure 0.05Mpa, incubation time 22 hours.The complete fermented liquid that ferments is concentrated in vacuo to 45% of original volume through low-temperature negative-pressure, obtains bacterium concentrated solution.Add carrier: in concentrated solution, add the carrier mixing, mix; The weight ratio of concentrated solution and carrier is 0.5:1, and vehicle group becomes: CaCO
325 parts, 12 parts, dextrin.Fluidised bed drying, 50 ℃ of drying temperatures.
Substratum consists of: casein peptone 1%, beef extract 1%, yeast extract 0.5%, glucose 0.5%, sodium acetate 0.5%, citric acid diamines 0.2%, Tween800.1%, K2HPO40.2%, MgSO4.7H2O0.02%, MnSO4.H2O0.005%, CaCO32%, agar 1.5%, pH6.8.
Embodiment 2
Substantially with embodiment 1
Provide a kind of green micro-ecological composite fertilizer: parts by weight consist of: 5 parts of aspergillus niger cultures, 7 parts of colloid bud pole bacteria agents, 6 parts of phosphorus decomposing Bacillus megaterium microbial inoculums, subtilis culture 12, Aspergillus awamori culture 6,2 parts of plant lactobacillus agent.
The experiment of product effect
Selection experimental field and test design: tested 20 days-September 30 March in 2009 and carry out in Ba Bao village, dapple pond town, Yanchi county Ningxia.
Experimental plot reaches 10 mu of field maize plantings, uses 0.7 kilogram every mu of product of the present invention respectively when plantation, emerges and by loose ground mode, uses 0.6 kilogram of invention product about 1 month, and control group is used conventional fertilizer.
Invention product is used corn field corn yield to reach 650 kilograms, and control group reaches 510 kilograms; This plot was at the 2nd year plant spring wheat, and spring wheat production has reached 400 kilograms, than control group per unit area yield, has improved 20%.And experimental plot Soil structure is good, without bulk and hardening.
Claims (6)
1. a preparation method for the micro-ecological composite fertilizer of green, comprises the steps: aspergillus niger culture, colloid bud pole bacteria agent, phosphorus decomposing Bacillus megaterium microbial inoculum, subtilis culture, Aspergillus awamori culture, plant lactobacillus agent to be combined as in proportion micro-ecological composite fertilizer.
2. the preparation method of green micro-ecological composite fertilizer according to claim 1, it is characterized in that, parts by weight consist of: aspergillus niger culture 5-10 part, colloid bud pole bacteria agent 3-8 part, phosphorus decomposing Bacillus megaterium microbial inoculum 3-8 part, subtilis culture 5-12, Aspergillus awamori culture 4-7, plant lactobacillus agent 2-4 part; Aspergillus niger (Aspergillusniger) preserving number is CGMCCNO.7927.
3. the preparation method of green micro-ecological composite fertilizer according to claim 1, it is characterized in that, described subtilis culture preparation method is as follows: from inclined-plane switching, cultivate subtilis, seed liquor after spreading cultivation is step by step transferred in fermentor tank, and the complete fermented liquid that ferments obtains subtilis culture through Plate Filtration, after dry.
4. the preparation method of green micro-ecological composite fertilizer according to claim 1, is characterized in that, described subtilis (Bacillussubtilissubsp) is CGMCC7926.
5. the method for effecting a permanent cure of green micro-ecological composite fertilizer according to claim 2, it is characterized in that, green micro-ecological composite fertilizer parts by weight consist of: 6 parts of aspergillus niger cultures, 5 parts of colloid bud pole bacteria agents, 5 parts of phosphorus decomposing Bacillus megaterium microbial inoculums, subtilis culture 10, Aspergillus awamori culture 6,3 parts of plant lactobacillus agent.
6. the preparation method of green micro-ecological composite fertilizer according to claim 2, it is characterized in that, green micro-ecological composite fertilizer parts by weight consist of: 5 parts of aspergillus niger cultures, 7 parts of colloid bud pole bacteria agents, 6 parts of phosphorus decomposing Bacillus megaterium microbial inoculums, subtilis culture 12, Aspergillus awamori culture 6,2 parts of plant lactobacillus agent.
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| CN104355689A (en) * | 2014-09-30 | 2015-02-18 | 天津天绿健科技有限公司 | Micro-ecological fertilizer and preparing method thereof |
| CN104450573A (en) * | 2014-12-02 | 2015-03-25 | 大丰市佳丰油脂有限责任公司 | Compound ecological microbial agent containing microbial polysaccharide and functional microbes |
| CN104692601A (en) * | 2015-02-12 | 2015-06-10 | 绿野工程私人有限公司 | Oil sludge and sand recycling method and system |
| CN105924238A (en) * | 2016-04-20 | 2016-09-07 | 义乌市锦钰信息科技有限公司 | Microbial compound fertilizer and preparation method thereof |
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| CN1140159A (en) * | 1996-04-29 | 1997-01-15 | 刘文治 | Method for producing organic and inorganic mixed microbe fertilizer |
| CN101914447A (en) * | 2010-07-31 | 2010-12-15 | 大连三科生物工程有限公司 | Complex microbial inoculum 707 and preparation method and application thereof |
| CN102168045A (en) * | 2010-12-24 | 2011-08-31 | 北京科为博生物科技有限公司 | Bacillus subtilis preparation and preparation method thereof |
| CN102277349A (en) * | 2011-07-24 | 2011-12-14 | 李绩 | Production method of biological compound enzyme |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104341183A (en) * | 2014-09-22 | 2015-02-11 | 北京科慧通智慧科技有限公司 | Preparation method of biofertilizer |
| CN104355689A (en) * | 2014-09-30 | 2015-02-18 | 天津天绿健科技有限公司 | Micro-ecological fertilizer and preparing method thereof |
| CN104450573A (en) * | 2014-12-02 | 2015-03-25 | 大丰市佳丰油脂有限责任公司 | Compound ecological microbial agent containing microbial polysaccharide and functional microbes |
| CN104692601A (en) * | 2015-02-12 | 2015-06-10 | 绿野工程私人有限公司 | Oil sludge and sand recycling method and system |
| CN105924238A (en) * | 2016-04-20 | 2016-09-07 | 义乌市锦钰信息科技有限公司 | Microbial compound fertilizer and preparation method thereof |
| CN109679870A (en) * | 2019-01-07 | 2019-04-26 | 山东省林业科学研究院 | A kind of biological organic fertilizer and preparation method thereof with water conservation drought resisting function |
| CN109679870B (en) * | 2019-01-07 | 2019-10-29 | 山东省林业科学研究院 | A kind of biological organic fertilizer and preparation method thereof with water conservation drought resisting function |
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