CN103626531B - Compound fertilizer and preparation method thereof - Google Patents
Compound fertilizer and preparation method thereof Download PDFInfo
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- CN103626531B CN103626531B CN201310589442.XA CN201310589442A CN103626531B CN 103626531 B CN103626531 B CN 103626531B CN 201310589442 A CN201310589442 A CN 201310589442A CN 103626531 B CN103626531 B CN 103626531B
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- 239000003337 fertilizer Substances 0.000 title claims abstract description 57
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 150000001875 compounds Chemical class 0.000 title abstract description 7
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 25
- 241001513093 Aspergillus awamori Species 0.000 claims abstract description 13
- 239000000084 colloidal system Substances 0.000 claims abstract description 12
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 11
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 7
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 7
- 241000894006 Bacteria Species 0.000 claims description 26
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 13
- 229910052698 phosphorus Inorganic materials 0.000 claims description 13
- 239000011574 phosphorus Substances 0.000 claims description 13
- 238000001035 drying Methods 0.000 claims description 11
- 241000186660 Lactobacillus Species 0.000 claims description 10
- 239000002131 composite material Substances 0.000 claims description 10
- 229940039696 lactobacillus Drugs 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 10
- 241000194107 Bacillus megaterium Species 0.000 claims description 8
- 239000002068 microbial inoculum Substances 0.000 claims description 8
- 230000007480 spreading Effects 0.000 claims description 3
- 238000003892 spreading Methods 0.000 claims description 3
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- 230000012010 growth Effects 0.000 abstract description 2
- 229940072205 lactobacillus plantarum Drugs 0.000 abstract description 2
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- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
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- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 244000241257 Cucumis melo Species 0.000 description 1
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- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- KIWBPDUYBMNFTB-UHFFFAOYSA-N Ethyl hydrogen sulfate Chemical compound CCOS(O)(=O)=O KIWBPDUYBMNFTB-UHFFFAOYSA-N 0.000 description 1
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- 241000237502 Ostreidae Species 0.000 description 1
- 102000015439 Phospholipases Human genes 0.000 description 1
- 108010064785 Phospholipases Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
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- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 1
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- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
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- IXORZMNAPKEEDV-OBDJNFEBSA-N gibberellin A3 Chemical class C([C@@]1(O)C(=C)C[C@@]2(C1)[C@H]1C(O)=O)C[C@H]2[C@]2(C=C[C@@H]3O)[C@H]1[C@]3(C)C(=O)O2 IXORZMNAPKEEDV-OBDJNFEBSA-N 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
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- 229910052739 hydrogen Inorganic materials 0.000 description 1
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- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 description 1
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fertilizers (AREA)
Abstract
The invention discloses a compound fertilizer and a preparation method thereof. The compound fertilizer is composed of the following ingredients in parts by weight: 5-10 parts of a yeast agent, 5-10 parts of a colloid bacillus microbial agent, 5-10 parts of a phosphors-resolving bacillus-subtilis microbial agent, 10-15 parts of bacillus subtilis culture, 1-5 parts of aspergillus awamori culture and 1-4 parts of a lactobacillus plantarum agent. The fertilizer provided by the invention is rich in nutrition and high in organic matter content. The compound fertilizer provided by the invention is capable of meeting the production requirements of organic food. The fertilizer provided by the invention is formed by fermenting compound microorganisms, and thus is capable of improving the micro-ecological environment of soil and proving good conditions for the growth of crops.
Description
Technical field
The present invention relates to a kind of composite fertilizer, belong to agricultural fertilizer technical field.
Background technology
The volume increase of application to agricultural of chemical fertilizer plays an important role, but due to long-term application chemical fertilizer, fertilizer is not enough, various nutrients ratio imbalance in soil, causes the destruction that farmland ecological environment, soil physical and chemical property and soil microflora are subject in various degree.Also have impact on the quality of agricultural-food to a certain extent, " fruit is sweetless, and melon is not fragrant " is exactly the performance of this situation simultaneously.Pollution by chemical fertilizer has become the large public hazards in the world today one.Agricultural experts unanimously appeal, Environment control, must reduce fertilizer amount.Expert, the new academician of China Agricultural University Chen Wen think, the excessive chemical fertilizer of using of current China arrives the limit.For nitrogenous fertilizer, the average amount of application of China is nearly 3 times, more than 8 times of Australia of the U.S., which results in that Soil structure is destroyed, a series of serious problems such as nature biotechnology diversity and ecosystem stability reduction.Agricultural microbiology Professional Committee of Chinese Society for Microbiology director doctor Li Jun points out, widelys popularize microorganism fertilizer and coordinates chemical fertilizer application to be exactly one of solution to this problem.
Microorganism fertilizer a kind ofly causes farm crop to obtain the living microorganisms goods of specific fertilizer effect with microbial life activity and product thereof, it at culture fertility, improve chemical fertilizer utilization ratio, suppress the absorption of the objectionable impurities such as farm crop heavy metal and agricultural chemicals, have irreplaceable effect in utilizations of becoming thoroughly decomposed, raising quality of agricultural product etc. of purification and rehabilitating soil, promotion agricultural crop straw and municipal wastes.The efficacy exertion of microbial fertilizer is mainly by the synergism to traditional fertilizer, fertilizer, and to the improvement activation of soil, and the mode such as the physiological action of microorganism realizes.Therefore, microbial fertilizer has just had chemical fertilizer, fertilizer, the multiple fertility of beneficial organism bacterium and effect, is activated fertilizer nutrient, improves effect of fertilizer, Crop Improvement quality, promotes the desirable fertilizer of agricultural production efficiency.Microbial fertilizer is live body fertilizer, and a large amount of beneficial microorganism vital movements that its effect mainly contains by it have been come.Only have when these beneficial microorganism vital movements have been come.Only have under these beneficial microorganisms are in vigorous breeding and metabolic situation, Substance Transformation and beneficial metabolic product could constantly be formed.Therefore, whether beneficial microorganism kind in microbial fertilizer, vital movement vigorous is the basis of its validity, and unlike other fertilizer be by the form of the principal elements such as nitrogen, phosphorus, potassium and how many based on.Just because of microbial fertilizer is preparation of living, thus its fertilizer efficiency and number of viable, intensity and ambient environmental conditions closely related, comprise temperature, moisture, potential of hydrogen, nutritional condition and formerly live in indigenous microorganism repulsive interaction in soil and have certain influence.
China's microbial fertilizer industry is compared with other countries, have that Varieties is many, the feature of applied range, especially, in the product innovation be composited research and development and application microorganism and organic nutrient substance, microorganism and inorganic nutrient substance, maintain the leading position.But, due to China for a long time in microbial fertilizer research lack and drop into, the microbial fertilizer industry of China is still existed, and integral level is not high, technological innovation is not enough, quality product and effect show understable problem.Today of human consensus has been become, " bottleneck " that these problems have become microorganism fertilizer industry letter to be got through at agricultural sustainable development.
Summary of the invention
Technical problem to be solved by this invention overcomes the deficiencies in the prior art, provides a kind of composite fertilizer:
Parts by weight consist of: yeast agent 5-10 part, colloid bud pole bacteria agent 5-10 part, phosphorus decomposing Bacillus megaterium microbial inoculum 5-10 part, subtilis culture 10-15, Aspergillus awamori culture 1-5, plant lactobacillus agent 1-4 part.
Described subtilis culture preparation: cultivate subtilis from inclined-plane switching, the seed liquor after spreading cultivation step by step is transferred in fermentor tank, and the complete fermented liquid that ferments obtains subtilis culture after Plate Filtration, drying.
The preparation method of plant lactobacillus agent: from inclined-plane switching culturing plants Bacterium lacticum, the seed liquor after spreading cultivation step by step is transferred in fermentor tank, and the complete fermentation liquor low-temperature negative-pressure that ferments is concentrated in vacuo to 45% of original volume, obtains bacterium concentrated solution.Add carrier: in concentrated solution, add the carrier mixed, 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, drying temperature 50 DEG C.
Prepared by Aspergillus awamori culture: spawn culture, and solid fermentation is cultivated: spore liquid is inoculated in aspergillus oryzae solid state fermentation culture material, and 26-33 DEG C is cultured to mycelia and covers with culture material, and low temperature fluidized bed dry, pulverize dry thing;
This product using method is simple, every mu of ground usage quantity 0.3-1 kilogram, and cost is only 80-100 unit/mu, and seed dressing or front earth's surface of pouring water vegetative period are sprayed.
Beneficial effect
Through testing proof for many years, the conversion energy of soil organisms enzyme improve because of long-term partially to fertilize and the soil compaction caused, acidifying, saliferous problem, make sandy soil reunite, clay loosen, strengthen water conservation, fertilizer-preserving ability, biological enzymes a large amount of in soil has the ability of point phosphorus decomposing.But the inorganic fertilizer that life-time service is a large amount of and agricultural chemicals, make some probiotics become extinct and disappear.So we are manured into soil with appropriate compound bio-enzyme, the loosing soil of active layer can be impelled, increase soil fertility, build the good soil environment that is suitable for crop root growth.The conversion of soil organisms enzyme, reducing agriculture production cost, reducing the use of chemical fertilizer, recovering all can play significant effect in soil ecology soil fertility and effective raising crop yield etc., gives full play to the organic usefulness of soil.
Colloid bacillus cereus is also known as silicate bacteria, its key property is potassium, the silicon that can decomposite in the mineral such as feldspar, mica, also the phosphorus in phosphatic rock be can decomposite, and plant growth substance and multiple enzyme secreted, to strengthen the resistibility of crop to some diseases.
Endobacillary potassium, after thalline death, dissociates out, can be absorbed and used by plants again.As a kind of critical function bacterium in microbial fertilizer, soil available nitrogen, phosphorus content can be improved, improve the multiple effect such as crop yield and quality.
Rui Gu bio tech ltd, Baoding provides colloid bacillus cereus bacterium powder;
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 substance such as growth hormone material and Plant hormones regulators,gibberellins material, extraneous root strong sprout; Decomposing element silicon in soil, for plant utilization, makes the wax layer of plant thicken, and improves plant water keeping ability; Can promote that soil aggregate is formed, and keeps soil from packing together; Spoiled soil capillarity, prevents soil water evaporation.
Fertilizer nutrient of the present invention is enriched, and organic content is high.The need of production of Organic food can be met.
Fertilizer of the present invention is formed by compound microorganism ferments, can improve the micro-ecological environment of soil, for the growth of farm crop provides beneficial conditions.
Fertilizer of the present invention can strengthen the diseases and insect pests resistance of crop.
Fertilizer of the present invention can improve 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, soil aggregate can be improved, strengthen the physicals of soil and reduce the loss of soil particle, under certain conditions, soil ulmin can also be participated in and formed.So use microbial fertilizer can improve soil physical property, be conducive to increasing 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, and seed dressing or front earth's surface of pouring water vegetative period are sprayed.
Embodiment
Unless stated otherwise, technique means used in the present invention 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 only limited by claims.To those skilled in the art, under the prerequisite not deviating from essence of the present invention and scope, the various change carry out the reaction conditions in these embodiments, separation and Extraction condition or change also belong to protection scope of the present invention.
The following examples can make the present invention of those skilled in the art comprehend, but do not limit the present invention in any way.
Phosphorus decomposing Bacillus megaterium is a kind in Bacillus (Bacillus).Motion, forms pod membrane, aerobic.Acid is produced from glucose, also normal from pectinose and the acid of N.F,USP MANNITOL product.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, Hebei China Cangzhou City Canal Zone chin or cheek and Building A 1, international business affairs center 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 (be called for short CGMCC, address is: No. 3, No. 1, North Star West Road, Chaoyang District, city of BeiJing, China institute) on July 15th, 2013, and 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, 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 the positive.
Described subtilis (Bacillus subtilis) Li-2013-02 is preserved in the product Thermostable α-Amylase in this laboratory subtilis Li-2013 by a strain obtains through UV-LiCl-ethyl sulfate Mutation screening, and concrete screening step is as follows:
(1) preparation of bacteria suspension
The mono-bacterium colony of Li-2013 grown after plate streaking is separated is accessed in seed culture medium, 100r/min, after 40 DEG C of cultivation 12h, after getting 1mL medium centrifugal, use brine twice, and resuspended with 9mL physiological saline.
(2) UV-LiCl-ethyl sulfate complex mutation
Bacteria suspension is placed in aseptic flat board, is 30cm in distance, stirs and irradiate 100s under the ultraviolet lamp of power 15w.Bacterium liquid through irradiating is coated lithium chloride flat board after gradient dilution, and contrasts to be coated with flat board without the bacterium liquid dilution of uv irradiating.Above-mentioned coating is dull and stereotyped uniformly, wrap with the cloth of black or newspaper, put 40 DEG C and cultivate 48h, the flat board growing bacterium colony filters out hydrolysis circle choose preserve to inclined-plane with colony diameter ratio the maximum, bacteria suspension is mixed with after purifying, fully mix with ethyl sulfate stoste after gradient dilution, and in 40 DEG C of concussion process 40min, the bacterium liquid processed is coated lithium chloride flat board after gradient dilution.
(3) primary dcreening operation of high-yield strains
Above-mentioned coating is dull and stereotyped uniformly, and put 40 DEG C and cultivate 48h, on the flat board growing bacterium colony, primary dcreening operation goes out hydrolysis circle and chooses preserve to inclined-plane with colony diameter ratio the greater, obtains three strain bacterium Li-2013-01, Li-2013-02, Li-2013-03 after purifying
(4) shake flask fermentation sieves again
By the three strain bacterium Li-2013-01 obtained, Li-2013-02, Li-2013-03 carry out shake flask fermentation in the 250mL shaking flask containing 30mL fermention medium, seed inoculum size 10% (V/V), 40 DEG C, 100r/min cultivates 72h, centrifuging and taking fermented supernatant fluid obtains crude enzyme liquid.
(5) enzyme activity determination
The definition of Mei Huo unit: 1mL crude enzyme liquid, in 105 DEG C, under pH 4.2 condition, 1min liquefies 1mg Zulkovsky starch, is 1 enzyme activity unit, represents with U/mL.
After measured, bacterial strain Li-2013-02 is stable most superior strain, and enzyme is lived and reached 30000U/mL, to live raising 1.6 times than original strain enzyme.
Described lithium chloride is dull and stereotyped: starch 1%, 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%, NaCl 1%.
Described fermention medium: Semen Maydis powder 5% ~ 15%, soybean cake powder 4% ~ 10%, (NH) 2SO40.4%, K2HPO40.8%, CaCl20.2%.
Described shake flask culture conditions: this bacterium in the 250mL shaking flask containing 30mL fermention medium, inoculum size 10% (V/V), 100r/min, 40 DEG C of fermentation culture 72h.
Fermented by bacterial strain Li-2013-02 and obtain 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 DEG C, and the temperature stability of preserving below 110 DEG C is better, and more than 115 DEG C to preserve long-time temperature stability poor.
(2) this enzyme optimal reaction pH value is 4.2.High enzyme vigor is all had, the enzyme complete stability alive when pH value is 3.0 between pH value 3.0-7.0.
(3) enzymic activity: by mutant strain Li-2013-02 provided by the present invention, the Thermostable 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 plant height product Thermostable α-Amylase, and this bacterial strain has acidproof, thermotolerance by force, produces the feature that enzyme activity is high.
2, this bacterial strain is had to produce the Thermostable α-Amylase enzyme activity of gained up to 30000-35000u/ml; Applicable temperature scope is 25-115 DEG C, optimal reactive temperature 110 DEG C, at 110 DEG C of enzymes complete stability alive; Being suitable for pH value in reaction scope is 3.0-7.0, the enzyme complete stability alive when pH value is 3.0, optimal reaction pH value is 4.2, higher than existing Thermostable α-Amylase enzyme activity, enzyme effect optimum pH wide scope, resistance to temperature is high, is particularly suitable for that temperature of reaction is high, liquefaction process and Mashing process the industrialization demand of depositing.
Embodiment 1
A kind of composite fertilizer, parts by weight consist of: yeast agent 8 parts, colloid bud pole bacteria agent 8 parts, phosphorus decomposing Bacillus megaterium microbial inoculum 8 parts, subtilis culture 12, Aspergillus awamori culture 3, plant lactobacillus agent 3 parts.
The bacterial classification adopted is as follows:
Subtilis (Bacillus subtilis subsp) CGMCC7926
Plant lactobacillus (Lactobacillus plantarum) CICC20764
Aspergillus awamori (Aspergillus awamori) CGMCC3.6484
S. cervisiae selects cicc1526, cicc1413, and bacterial classification is all purchased from Chinese industrial Culture Collection.
The production method of yeast microbial inoculum is see Xiao Dongguang work " production of Active Dry Yeast and utilisation technology ", and the Inner Mongol People's Press publishes for 1994.
The preparation method of Aspergillus awamori microbial inoculum:
Technical scheme is as follows:
Slant strains activation culture: be transferred on slant medium by Aspergillus awamori slant strains, cultivates 3 days for 27 DEG C.
Solid first order seed is cultivated: the access of picking Aspergillus awamori slant strains is equipped with in 500 milliliters of triangular flasks of 100 grams of substratum and is carried out seed culture, cultivates 3 days for 30 DEG C.
Solid secondary seed is cultivated: stirred by above-mentioned cultured solid first order seed and carry out seed culture, culture condition for adding in 5000 milliliters of triangular flasks that 1000 grams of substratum are housed after fragment: cultivate 3 days for 30 DEG C.
Solid fermentation is cultivated: pulverized by second-level shake flask seed, add in the fermentation vat or pallet that sterilising medium is housed and mix rear cultivation, and bent material culture temperature controls at 26-35 DEG C, humidity 80-90%, every 10 hours stirrings once, and incubation time 5-7 days; The cultivation of solid koji material adopts conventional bent material culture technique; Treat culture material cover with mycelia can terminate cultivate, substratum in advance through high temperature steaming sterilising treatment, sterilising conditions control temperature 121 DEG C, 1 hour time.
Drying and crushing: fermentation ends culture material carries out drying on fluidized-bed or other drying plants, drying temperature controls, at 60 DEG C, to be dried to moisture content below 10%, then to be pulverized by solid culture medium, and crushing material aperture is more than 60 orders.
Substratum forms: solid material: wheat bran 80%, 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 accessed in 500 ml shake flasks, substratum loading amount 100 milliliters, rotary shaker 180 revs/min, culture temperature 30 DEG C, incubation time 24 hours;
(2) secondary seed cultivate: by first order seed according to 10% inoculum size access in 500 milliliters of secondary seed shaking flasks, 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 flask with 10% inoculum size, substratum loading amount 1000 milliliters, rotary shaker 100 revs/min, culture temperature 30 DEG C, incubation time 24 hours;
(4) first class seed pot is cultivated: the first class seed pot being 150L with 10% inoculum size access cubic capacity by three grades of seeds, fermention medium loading amount 100L, culture temperature 28 DEG C, stirring velocity 100 revs/min, 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 is accessed cubic capacity with 10% inoculum size, fermention medium loading amount 1 ton, culture condition culture temperature 28 DEG C, stirring velocity 100 revs/min, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa, incubation time 24 hours.
Substratum forms: glucose 6%, yeast extract 1%, peptone 0.2%, CaCO3 1%, pH6.8.
The preparation method of plant lactobacillus agent:
(1) first order seed is cultivated: accessed by plant lactobacillus bacterial classification in 500 ml shake flasks, substratum loading amount 100 milliliters, culture temperature 30 DEG C, incubation time 24 hours;
(2) secondary seed cultivate: by first order seed according to 10% inoculum size access in 500 milliliters of secondary seed shaking flasks, 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 flask with 10% inoculum size, substratum loading amount 1000 milliliters, culture temperature 30 DEG C, incubation time 24 hours;
(4) first class seed pot is cultivated: the first class seed pot being 150L with 5% inoculum size access cubic capacity by three grades of seeds, fermention medium loading amount 100L, culture temperature 30 DEG C, 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 is accessed cubic capacity with 5% inoculum size, fermention medium loading amount 2 tons, culture condition culture temperature 30 DEG C, tank pressure 0.05Mpa, incubation time 22 hours.The complete fermentation liquor low-temperature negative-pressure that ferments is concentrated in vacuo to 45% of original volume, obtains bacterium concentrated solution.Add carrier: in concentrated solution, add the carrier mixed, 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, drying temperature 50 DEG C.
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%, Tween 80 0.1%, K2HPO4 0.2%, MgSO4.7H2O0.02%, MnSO4.H2O 0.005%, CaCO3 2%, agar 1.5%, pH6.8.
Embodiment 2
Basic with embodiment 1
A kind of composite fertilizer, parts by weight consist of: yeast agent 9 parts, colloid bud pole bacteria agent 7 parts, phosphorus decomposing Bacillus megaterium microbial inoculum 7 parts, subtilis culture 13, Aspergillus awamori culture 2, plant lactobacillus agent 2 parts.
Product effect experimental
Selection experimental field and test design: test and carry out in Ba Bao village, dapple pond town, Yanchi county Ningxia 27 days-September 30 April in 2009.
Experimental plot reaches field maize planting 10 mu, using product every mu of the present invention 0.5 kilogram respectively, emerging about 1 month and using invention product 0.5 kilogram by loose ground mode when planting, and control group uses common fertilizer.
Invention product uses corn field corn yield to reach 650 kilograms, and control group reaches 500 kilograms; The use of invention product makes corn per mu yield improve 30%.This plot was at the 2nd year plant spring wheat, and spring wheat production reaches 400 kilograms, improves 20% than control group per unit area yield.And experimental plot Soil structure is good, without bulk with harden.
Claims (4)
1. a composite fertilizer, parts by weight consist of: yeast agent 5-10 part, colloid bud pole bacteria agent 5-10 part, phosphorus decomposing Bacillus megaterium microbial inoculum 5-10 part, subtilis culture 10-15, Aspergillus awamori culture 1-5, plant lactobacillus agent 1-4 part, described subtilis (Bacillus subtilis) preserving number is CGMCC No.7926.
2. a kind of composite fertilizer according to claim 1, described subtilis culture preparation: cultivate subtilis from inclined-plane switching, seed liquor after spreading cultivation step by step is transferred in fermentor tank, and the complete fermented liquid that ferments obtains subtilis culture after Plate Filtration, drying.
3. a kind of composite fertilizer according to claim 1, parts by weight consist of: yeast agent 8 parts, colloid bud pole bacteria agent 8 parts, phosphorus decomposing Bacillus megaterium microbial inoculum 8 parts, subtilis culture 12, Aspergillus awamori culture 3, plant lactobacillus agent 3 parts.
4. a kind of composite fertilizer according to claim 1, parts by weight consist of: yeast agent 9 parts, colloid bud pole bacteria agent 7 parts, phosphorus decomposing Bacillus megaterium microbial inoculum 7 parts, subtilis culture 13, Aspergillus awamori culture 2, plant lactobacillus agent 2 parts.
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| CN105234168A (en) * | 2015-11-20 | 2016-01-13 | 福建康德贝生物科技有限公司 | Application of bacillus subtilis |
| CN110218683B (en) * | 2019-06-27 | 2020-06-26 | 中国科学院西北生态环境资源研究院 | Microbial composition, microbial preparation and method for improving soil in sand area |
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| CN102277343A (en) * | 2011-07-24 | 2011-12-14 | 李绩 | Biological composite enzyme |
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| 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 |
| CN102153380A (en) * | 2010-12-15 | 2011-08-17 | 沈阳科丰牧业科技有限公司 | Method for preparing composite probiotic spraying agent |
| CN102153382A (en) * | 2010-12-15 | 2011-08-17 | 沈阳科丰牧业科技有限公司 | Composite microbial foliar fertilizer spraying agent and using method thereof |
| CN102277343A (en) * | 2011-07-24 | 2011-12-14 | 李绩 | Biological composite enzyme |
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