CN102786934A - Probiotic soil conditioner and production method thereof - Google Patents
Probiotic soil conditioner and production method thereof Download PDFInfo
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- CN102786934A CN102786934A CN2012102994830A CN201210299483A CN102786934A CN 102786934 A CN102786934 A CN 102786934A CN 2012102994830 A CN2012102994830 A CN 2012102994830A CN 201210299483 A CN201210299483 A CN 201210299483A CN 102786934 A CN102786934 A CN 102786934A
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- seed liquor
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- probiotic
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- 239000006041 probiotic Substances 0.000 title claims abstract description 42
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 41
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 41
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 38
- 239000003516 soil conditioner Substances 0.000 title claims abstract description 32
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 45
- 238000000034 method Methods 0.000 claims abstract description 23
- 238000000855 fermentation Methods 0.000 claims abstract description 21
- 230000004151 fermentation Effects 0.000 claims abstract description 21
- 241000894006 Bacteria Species 0.000 claims description 45
- 241000194107 Bacillus megaterium Species 0.000 claims description 36
- 230000001954 sterilising effect Effects 0.000 claims description 22
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 20
- 239000002609 medium Substances 0.000 claims description 20
- 238000011534 incubation Methods 0.000 claims description 18
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- 238000002360 preparation method Methods 0.000 claims description 16
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- 108010064696 N,O-diacetylmuramidase Proteins 0.000 description 1
- 108010025915 Nitrite Reductases Proteins 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
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- CLKOFPXJLQSYAH-ABRJDSQDSA-N bacitracin A Chemical compound C1SC([C@@H](N)[C@@H](C)CC)=N[C@@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]1C(=O)N[C@H](CCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2N=CNC=2)C(=O)N[C@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCCCC1 CLKOFPXJLQSYAH-ABRJDSQDSA-N 0.000 description 1
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a production method of a probiotic soil conditioner. The production method comprises the steps of: (1) respectively inoculating huge bacillus, bacillus subtilis and bacillus licheniformis into a seed cultivation medium to cultivate and obtaining respective seed liquid; (2) adding the cultivation medium for production into a fermentation tank to sterilize and cool; and (3) inoculating the bacillus seed liquid into the fermentation tank to cultivate under an oxygen-leading agitating condition, inoculating the bacillus subtilis seed liquid and bacillus licheniformis seed liquid to cultivate, and obtaining the probiotic soil conditioner. The invention also discloses the probiotic soil conditioner obtained by the method. The method disclosed by the invention is scientific and reasonable, simple and feasible in production process and strong in industrialization implementation, and has an important significance to the development of agricultural production.
Description
Technical field
The invention belongs to the agro-biological engineering technical field, relate to agricultural microorganism fertilizer or bio-organic fertilizer production technology, relate in particular to a kind of probiotic type soil conditioner and working method thereof.
Background technology
Last century five, the sixties, the use of chemical fertilizer and agricultural chemicals makes China's Agricultural Development obtain significant progress, has also brought huge benefit to the peasant.But a large amount of and excess of chemical fertilizer and agricultural chemicals is used, and has caused irreversible injury for soil, environment and even the ecosystem.Such as soil compaction; Soil water retention power descends; Fertility is unbalanced in the soil, and fertility descends; The input and output ratio descends.Even more serious is that these chemical substances can cause the animal-plant material hazardous and noxious substances to exceed standard at the plant-animal body accumulation, have a strong impact on quality of agricultural product.Meanwhile, residual chemical substances is through the ground water regime of soil, and the polluted water resource causes irreversible destruction in ecosystem circulation.
The infringement that chemical fertilizer causes is well-known, and in the case, the application of bio-feritlizer becomes the focus in the agricultural development.Wherein microbial fertilizer is a kind of new-type fertilizer that grows up nearly decades.Microbial fertilizer is meant the particular product that contains living microorganism.The major advantage of microbial fertilizer be can improve the soil, free from environmental pollution, have no side effect, be the desirable fertilizer of producing " green food ".Microbial fertilizer is applied to produce, can promotes soil fertility, assist plant absorbing nutrition, the disease-resistant and drought-resistant ability of enhancement of plant, save energy reduces production costs, and reduces environmental pollution.
Bacillus megaterium (Bacillus megaterium) is the short endophytic bacteria of a kind of root system of plant, also is the bacterial classification commonly used in the microbial fertilizer.It is the short potassium bacterium of a kind of phosphorus decomposing, and ADSORPTION STATE organophosphorus, the inorganic phosphorus that can't directly utilize plant in Yelkin TTS, the soil have tangible Decomposition, can increase soil fertility, and fixed nitrogen is had synergism, promote crop yield.Bacillus megaterium can reduce organonitrogen, the phosphorus in the water body, and the prussiate in the trade effluent of can also to a certain degree degrading etc. play a role to the eubiosis in the water body environment.In addition, bacillus megaterium can produce nitrite reductase, reduces nitrite content in the food.So the application prospect of bacillus megaterium is boundless.
Subtilis (Bacillus substilus) is a kind of gram positive bacterium of being regarded as GRAS (It is generally accepted safety) by FDA Food and Drug Administration (FDA); Extensively be present in nature; It is the dominant strain of the little ecology of plant rhizosphere; Can secrete the multiple secondary metabolite that plant pathogenic fungi is had antagonistic action; Like subtilyne, polymyxin, nystatin, linear gramicidins isoreactivity material, the conditioned pathogen of pathogenic bacterium or autogenous infection there is the obvious suppression effect; Synthetic various active enzyme and organic acid, VITAMINs, amino acids material in while self process of growth; Have the promotion plant growth, decompose in the release soil and combine nutrient, reduce soil pH; Improving advantages such as crop resistance, is a kind of biocontrol microorganisms that has using value.
Bacillus licheniformis (Bacillus licheniformis) is a kind of gram positive bacterium, and it is very extensive to distribute at occurring in nature, and physiological property is also rich and varied, is one of soil and plant microecology dominant population.Can produce the endosporic chmosynthetic heterotrophs bacterium of resistance under certain condition.Can produce peptide class antagonistic substance, thus the called after bacitracin, be a kind of strong sterilant, ability strongly inhibited G+ bacterium also has stronger restraining effect to part G-bacterium simultaneously.It can produce multiple microbiotic, antibacterial protein, small molecules bacteriocin, N,O-Diacetylmuramidase etc.; Multiple animal and plant and human pathogenic bacteria are all played the good restraining effect; And Bacillus licheniformis also has very strong proteolytic enzyme, lypase, amylases activity; External also have report to point out that Bacillus licheniformis can produce the bacterinertness material of one type of toluylic acid.Bacillus licheniformis is the most ripe in the research aspect medical, has formed more complete production technique, and also comparatively ripe in the living application of the whole intestines of medical commodity.But the research at aspects such as the processing that prevents corps diseases, feed, environmental pollution improvements is in the stage at the early-stage, and development space will be boundless.
Hindering at present the bottleneck problem that the microbiobacterial agent industrial or agricultural uses is, utilizes traditional microbial fermentation processes, and the microbiobacterial agent living bacteria count of acquisition is on the low side; Bacterial classification is single, and function singleness is applied to DeGrain in the industrial and agricultural production; If improve the living bacteria count amount, method such as need freeze-drying, concentrate, the input of costs such as equipment is higher; The finished product price is high, is not easy the person of being used and accepts; Be made into particle and use, bake out temperature is high during granulation, causes the viable bacteria mass mortality, and effect is poor.
National inventing patent: (publication number: CN1397521A) disclose a kind of liquid dosage form, contained azotobacter chroococcum, bacillus megaterium and silicate bacteria, the living bacteria count amount is 800,000,000/milliliter for a kind of liquid composite microbic bacterial fertilizer and working method thereof.In this invention, directly the liquid concentrator with three kinds of microbial inoculums mixes with nutritive substance by a certain percentage, and packing forms product.Problem is how the liquid concentrator of three kinds of microbial inoculums obtains so, and how much viable bacteria concentration of microbial inoculum can reach, and cause this patent of invention exploitativeness not strong, and wherein number of viable is also on the low side, and result of use can be affected.Another national inventing patent: the multielement compound microbe bacterial fertilizer (patent No.: ZL200610134541.9), contain 10 kinds of mikrobes, inoculate altogether 5 times, cultivate 52h, obtain the finished product.The problem one of this invention is to inoculate repeatedly to cause bacterium liquid to pollute easily, in case contaminated bacteria is bred in a large number, certainly will influence the vigor and the quantity of target bacterial classification, causes product quality problem; The 2nd, incubation time is long, obtains the fermentation that last bacterium liquid needs 52h, and time and energy consumption cost are all higher.
To the problems referred to above, the present invention provides a kind of probiotic type soil conditioner and working method thereof, and under the less energy-consumption condition, the short period of time fermentation obtains highdensity probiotic type composite microbic bacterial fertilizer.
Summary of the invention
The present invention provides a kind of probiotic type soil conditioner and working method thereof, and it is low to solve present microorganism bacterial manure field bacteria suspension concentration, and bacterial classification is single, function singleness, the problem of effect difference.
The present invention realizes through following technical scheme:
A kind of working method of probiotic type soil conditioner comprises:
(1) produces preparation: bacillus megaterium, subtilis, Bacillus licheniformis are inserted seed culture medium respectively cultivate with the probiotic bacterium seed liquor; Obtain bacillus megaterium seed liquor, subtilis seed liquor and Bacillus licheniformis seed liquor; Wherein the incubation time of bacillus megaterium is 8-12h, and the incubation time of subtilis and Bacillus licheniformis is 6-10h;
(2) produce preparation and sterilization: will produce with in the substratum adding fermentor tank, and sterilize 30-45 minute, and be cooled to 30-35 ℃ for 115-121 ℃ with substratum;
(3) inoculation and fermentation: according to using the ratio access fermentor tank of the volume ratio of substratum as 2-5% with said production, at 29-33 ℃, logical oxygen agitation condition is cultivated 10-18h down with said bacillus megaterium seed liquor; Then leavening temperature is adjusted to 33-36 ℃; Insert said subtilis seed liquor and Bacillus licheniformis seed liquor; Add simultaneously with said production and use the substratum TV, continue to cultivate 7-16h, obtain said probiotic type soil conditioner than concentrated sterilising medium as 4-10%.
Concrete operations are following:
(1) produce preparation with the probiotic bacterium seed liquor: with beef-protein medium as the seed fermentation substratum; Bacillus megaterium, subtilis, Bacillus licheniformis are inserted seed culture medium respectively to be cultivated; Obtain bacillus megaterium seed liquor, subtilis seed liquor and Bacillus licheniformis seed liquor; Wherein the incubation time of bacillus megaterium is 8-14h, and the incubation time of subtilis and Bacillus licheniformis is 6-12h; Subsequent use.
As the preferred seed incubation time, bacillus megaterium seed culture 10h, subtilis and Bacillus licheniformis seed are cultivated 8h respectively.
(2) produce preparation and sterilization: will produce with in the substratum adding fermentor tank, and sterilize 30-45 minute, and be cooled to 30-35 ℃ for 115-121 ℃ with substratum;
Said production is following with the prescription of substratum, and every liter of production is with containing in the substratum: sucrose 5-15g, lactose 5-12g, SANMALT-S 1-5g, Zulkovsky starch 1-5g, peptone 5-15g, yeast extract paste 2-9g, urea 1-5g, NaCl3-8g, K
2SO
40.4-1.0g, K
2HPO
40.5-1.5g, MgSO
47H
2O 2-3.5g, CaCl
20.8-1.5g, CaCO
34-6.5g surplus is a water; PH7.5-8.2.
As preferably, it is that every liter of production is with containing in the substratum that culture medium prescription is used in said production: sucrose 9-11g, lactose 7-8g, SANMALT-S 2-3g, Zulkovsky starch 2-3g, peptone 9-11g, yeast extract paste 5-6g, urea 2-3g, NaCl5-6g, K
2SO
40.6-0.75g, K
2HPO
40.9-1.1g, MgSO
47H
2O2.5-2.75g, CaCl
21.0-1.2g, CaCO
35-5.25g surplus is a water; PH7.9-8.1.
Preferred culture medium can provide enough carbon source, nitrogenous source and inorganic salt to growth, simultaneously the CaCO through adding
3PH in the control fermenting process was unlikely to low, the CaCl of adding
2, have the thalli growth of promotion simultaneously and produce the gemma effect, make that the product living bacteria count that obtains is high, the spore amount is big, guarantees post-production, and is applied to the good result in the industrial and agricultural production.
(3) inoculation and fermentation: according to using the ratio access fermentor tank of the volume ratio of substratum as 2-5% with said production, at 29-33 ℃, logical oxygen agitation condition is cultivated 10-18h down with said bacillus megaterium seed liquor; Then leavening temperature is adjusted to 33-37 ℃; Insert said subtilis seed liquor and Bacillus licheniformis seed liquor; Add simultaneously with said production and use the substratum TV, continue to cultivate 7-16h, obtain said probiotic type soil conditioner than concentrated sterilising medium as 4-10%;
The said subtilis seed liquor and the said Bacillus licheniformis seed liquor volume ratio that insert are 1:0.25-4; Said subtilis seed liquor is used culture volume than being 4-8% with the access total amount of said Bacillus licheniformis seed liquor with said production;
The said concentrated bactericidal nurishing base that adds consists of: every liter of concentrated sterilising medium contains: sucrose 30-50g, lactose 30-40g, SANMALT-S 5-15g, Zulkovsky starch 5-15g, peptone 25-35g, yeast extract paste 20-30g, NaCl 5-10g, K
2HPO
40.8-1.8g, MgSO
47H
2O 2-4g, CaCO
35-15g, surplus is a water; PH7.5-8.2.
As preferably, said bacillus megaterium seed liquor inserts said production and uses the volume ratio of substratum to be 3-4%, and culture temperature is 30-31 ℃, and incubation time is 11-12h.
Said leavening temperature is adjusted to 33-37 ℃,, leavening temperature is adjusted to 34-35 ℃ as preferably.
As preferably; Said subtilis seed liquor is used culture volume than being 5-6% with the access total amount of said Bacillus licheniformis seed liquor with said production; The said concentrated sterilising medium that adds uses the substratum TV than being 6-7% with said production, and the continuation incubation time is 9-11h.
As preferably, the said concentrated bactericidal nurishing base of adding consists of: every liter of concentrated bactericidal nurishing base contains: sucrose 40-42g, lactose 34-35g, SANMALT-S 7-8g, Zulkovsky starch 7-8g, peptone 29-31g, yeast extract paste 24-26g, NaCl 7-7.5g, K
2HPO
41.2-1.4g, MgSO
47H
2O 3-3.2g, CaCO
37-7.5g surplus is a water; PH7.8-8.0.
Under said inoculation and fermentation condition, cultivate, guarantee the quantity of three kinds of probiotic bacteriums in the final product that obtains, guarantee that simultaneously three kinds of genus bacillus all can produce a large amount of gemma, guarantee the effect of the finished product.
The present invention also provides a kind of probiotic type soil conditioner of working method preparation of described probiotic type soil conditioner, and living bacteria count adds up to hundred million/milliliter of 50-100 in the said probiotic type soil conditioner, and the gemma rate is at 90-100%.The living bacteria count of every kind of bacterium is greater than 200,000,000/milliliter.
Advantage of the present invention:
1. employed mikrobe is probiotic bacterium, and Product Green is nuisanceless, is suitable for green sustainable agriculture production and uses.
2. employing mixed fungus fermentation, the compound microbial bacterial manure-probiotic type soil conditioner of acquisition, living bacteria count is high, and powerful, effect is good.
3. the product application of probiotic type soil conditioner, improves outside the effect of edatope except having enhancing soil fertility in agriculture prodn, also has the powerful poisoning of separating, and anti continuous and alternative-year is anti-saline and alkaline, improves the characteristics of disease-resistant crops property, has a extensive future.
4. process step is simple, and is with short production cycle, and the bacterial manure of acquisition is energetic, reduces cost, and is suitable for batch production production.
Embodiment
Embodiment 1:
Produce preparation with the probiotic bacterium seed liquor: with beef-protein medium as the seed fermentation substratum; Bacillus megaterium, subtilis, Bacillus licheniformis are inserted seed culture medium respectively to be cultivated; Obtain bacillus megaterium seed liquor, subtilis seed liquor and Bacillus licheniformis seed liquor; Wherein the incubation time of bacillus megaterium is 12h, and the incubation time of subtilis and Bacillus licheniformis is 10h.
Produce preparation and sterilization with substratum: will produce with in the substratum adding fermentor tank, and produce as follows with the prescription of substratum, every liter of production is with containing in the substratum: sucrose 15g, lactose 12g; SANMALT-S 5g, Zulkovsky starch 5g, peptone 15g, yeast extract paste 9g; Urea 5g, NaCl 8g, K
2SO
41.0g, K
2HPO
41.5g, MgSO
47H
2O 3.5g, CaCl
21.5g, CaCO
36.5g surplus is a water; PH8.0.Sterilized 45 minutes, and be cooled to 34 ℃ for 121 ℃.
Inoculation and fermentation: the seed liquor that will cultivate the bacillus megaterium of 12h inserts fermentor tank according to using culture volume with production than the ratio that is 5%, and at 33 ℃, logical oxygen agitation condition is cultivation 16h down; Then leavening temperature is adjusted to 35 ℃, access subtilis seed liquor and Bacillus licheniformis seed liquor add simultaneously and production uses the substratum TV than the concentrated sterilising medium that is 10%, continue to cultivate 12h; The subtilis seed liquor and the Bacillus licheniformis seed liquor that insert, the two volume ratio is 3:7, inserting total amount is 8% with producing use culture volume ratio; The concentrated sterilising medium that adds consists of: every liter of concentrated sterilising medium contains: sucrose 50g, lactose 40g, SANMALT-S 15g, Zulkovsky starch 15g, peptone 35g, yeast extract paste 30g, NaCl 10g, K
2HPO
41.8g, MgSO
47H
2O4g, CaCO
315g, surplus is a water; PH8.2.
Obtain final probiotic type soil conditioner bacterial fertilizer products: the living bacteria count sum is 6,300,000,000/milliliter in the product, gemma rate 96.5%.The living bacteria count of every kind of bacterium is greater than 200,000,000/milliliter.
Embodiment 2:
Produce preparation with the probiotic bacterium seed liquor: with beef-protein medium as the seed fermentation substratum; Bacillus megaterium, subtilis, Bacillus licheniformis are inserted seed culture medium respectively to be cultivated; Obtain bacillus megaterium seed liquor, subtilis seed liquor and Bacillus licheniformis seed liquor; Wherein the incubation time of bacillus megaterium is 8h, and the incubation time of subtilis and Bacillus licheniformis is 6h.
Produce preparation and sterilization with substratum: will produce with in the substratum adding fermentor tank, and produce as follows with the prescription of substratum, every liter of production is with containing in the substratum: sucrose 8g, lactose 8g; SANMALT-S 2g, Zulkovsky starch 3g, peptone 6g, yeast extract paste 5g; Urea 3g, NaCl4g, K
2SO
40.44g, K
2HPO
40.8g, MgSO
47H
2O 2g, CaCl
20.8g, CaCO
34g, surplus is a water; PH8.2.Sterilized 45 minutes, and be cooled to 31 ℃ for 115 ℃.
Inoculation and fermentation: the seed liquor of bacillus megaterium that will cultivate 8h is according to inserting fermentor tank than 3% ratio with producing with culture volume, at 30 ℃, and cultivation 10h under the logical oxygen agitation condition; Then leavening temperature is adjusted to 33 ℃, access subtilis seed liquor and Bacillus licheniformis seed liquor add simultaneously and production uses the substratum TV than the concentrated sterilising medium that is 5%, continue to cultivate 8h; The subtilis seed liquor and the Bacillus licheniformis seed liquor that insert, the two volume ratio is 7:3, inserting total amount is 4% with producing use culture volume ratio; The concentrated sterilising medium that adds consists of: every liter of concentrated bactericidal nurishing base contains: sucrose 30g, lactose 30g, SANMALT-S 5g, Zulkovsky starch 5g, peptone 25g, yeast extract paste 20g, NaCl 5g, K
2HPO
40.8g, MgSO
47H
2O 2g, CaCO
35g, surplus is a water; PH8.2.
Obtain final probiotic type soil conditioner bacterial fertilizer products: the living bacteria count sum is 5,500,000,000/milliliter in the product, gemma rate 96.7%.The living bacteria count of every kind of bacterium is greater than 200,000,000/milliliter.
Embodiment 3:
Produce preparation with the probiotic bacterium seed liquor: with beef-protein medium as the seed fermentation substratum; Bacillus megaterium, subtilis, Bacillus licheniformis are inserted seed culture medium respectively to be cultivated; Obtain bacillus megaterium seed liquor, subtilis seed liquor and Bacillus licheniformis seed liquor; Wherein the incubation time of bacillus megaterium is 10h, and the incubation time of subtilis and Bacillus licheniformis is 8h.
Produce preparation and sterilization with substratum: will produce with in the substratum adding fermentor tank, and produce as follows with the prescription of substratum, every liter of production is with containing in the substratum: sucrose 10g, lactose 10g; SANMALT-S 3g, Zulkovsky starch 3g, peptone 10g, yeast extract paste 6g; Urea 2g, NaCl 5g, K
2SO
40.7g, K
2HPO
41.0g, MgSO
47H
2O 2.5g, CaCl
21.1g, CaCO
35g, surplus is a water; PH8.0.Sterilized 30 minutes, and be cooled to 32 ℃ for 121 ℃.
Inoculation and fermentation: the seed liquor of bacillus megaterium that will cultivate 10h is according to inserting fermentor tank than 4% ratio with producing with culture volume, at 31 ℃, and cultivation 13h under the logical oxygen agitation condition; Then leavening temperature is adjusted to 34 ℃, access subtilis seed liquor and Bacillus licheniformis seed liquor add simultaneously and production uses the substratum TV than the concentrated sterilising medium that is 5%, continue to cultivate 10h; The subtilis seed liquor and the Bacillus licheniformis seed liquor that insert, the two volume ratio is 1:1, inserting total amount is 6% with producing use culture volume ratio; The concentrated sterilising medium that adds consists of: every liter of concentrated bactericidal nurishing base contains: sucrose 40g, lactose 35g, SANMALT-S 10g, Zulkovsky starch 10g, peptone 30g, yeast extract paste 25g, NaCl 7g, K
2HPO
41.2g, MgSO
47H
2O3g, CaCO
310g, surplus is a water; PH8.0.
Obtain final probiotic type soil conditioner bacterial fertilizer products: the living bacteria count sum is 7,200,000,000/milliliter in the product, gemma rate 94.8%.The living bacteria count of every kind of bacterium is greater than 200,000,000/milliliter.
Embodiment 4:
Produce preparation with the probiotic bacterium seed liquor: with beef-protein medium as the seed fermentation substratum; Bacillus megaterium, subtilis, Bacillus licheniformis are inserted seed culture medium respectively to be cultivated; Obtain bacillus megaterium seed liquor, subtilis seed liquor and Bacillus licheniformis seed liquor; Wherein the incubation time of bacillus megaterium is 8h, and the incubation time of subtilis and Bacillus licheniformis is 8h.
Produce preparation and sterilization with substratum: will produce with in the substratum adding fermentor tank, and produce as follows with the prescription of substratum, every liter of production is with containing in the substratum: sucrose 8g, lactose 10g; SANMALT-S 4g, Zulkovsky starch 2g, peptone 8g, yeast extract paste 8g; Urea 3g, NaCl 7g, K
2SO
40.4g, K
2HPO
41.5g, MgSO
47H
2O 2g, CaCl
21.0g, CaCO
36.5g surplus is a water; PH7.8.Sterilized 42 minutes, and be cooled to 34 ℃ for 115 ℃.
Inoculation and fermentation: the seed liquor of bacillus megaterium that will cultivate 8h is according to inserting fermentor tank than 5% ratio with producing with culture volume, at 33 ℃, and cultivation 12h under the logical oxygen agitation condition; Then leavening temperature is adjusted to 34 ℃, access subtilis seed liquor and Bacillus licheniformis seed liquor add simultaneously and production uses the substratum TV than the concentrated sterilising medium that is 8%, continue to cultivate 8h; The subtilis seed liquor and the Bacillus licheniformis seed liquor that insert, the two volume ratio is 2:1, the access amount is 7% with producing use culture volume ratio; The concentrated sterilising medium that adds consists of: every liter of concentrated bactericidal nurishing base contains: sucrose 35g, lactose 32g, SANMALT-S 7g, Zulkovsky starch 7g, peptone 28g, yeast extract paste 20g, NaCl 5g, K
2HPO
41.5g, MgSO
47H
2O 3.3g, CaCO
38g, surplus is a water; PH8.1.
Obtain final probiotic type soil conditioner bacterial fertilizer products: the living bacteria count sum is 67.5 hundred million/milliliter in the product, gemma rate 96.7%.The living bacteria count of every kind of bacterium is greater than 200,000,000/milliliter.
Embodiment 5:
Field result of use: 2011-2012 bright villages of Xiangfang District in the Harbin City, Heilongjiang Province city respectively; Yong Fa village, Suihua City Golconda town, Heilongjiang Province; Lu He town, Qinggang County, Heilongjiang Province supports the army the village; Heilongjiang Province Hailun City truth potato planting cooperative society, ground such as Tong De village, the concentric township of Heilongjiang Province's Shuangcheng City are done experiment and are used probiotic type soil conditioner of the present invention, and supplying to study thing is corn, soybean, paddy rice, yam, tomato, muskmelon, capsicum.
1, TP:
Treatment group: the probiotic type soil conditioner stoste that fermentation obtains is diluted 300 times, with seed pouring plantation, every mu of consumption 800mL.Other field management are normally carried out.
Control group: do not use bacterial manure.Other field management are consistent with treatment group.
District's group is provided with at random, and treatment group and control group all are provided with 3 repetitions.Corn, soybean, every group of area 667m of paddy rice
2Yam, tomato, muskmelon, every group of area 200m of capsicum
2
2, testing index
In process of crop growth, observe crop pattern, growth potential, disease and pest situation, survey output at last.
3, result
Corn
Kind: elder generation beautiful 335.Sowing date: on May 1st, 2011.Harvest date: treatment group, on September 20th, 2011; Control group, on September 30th, 2011.The test place: Lu He town, Qinggang County, Heilongjiang Province supports the army the village; Soil characteristic: saltings.
Conclusion: the treatment group corn was emerged May 9, and it is neat to emerge, and the seedling growing way is healthy and strong; Than control group fast growth; And the thick leaf color depth of stem; Vegetative period, disease was few; Growth later stage and control group plant difference are little.Treatment group fruit fringe is big and long, and full seed does not have bald point.Treatment group was gathered 10 days than control group is early.The output investigation, treatment group output is 693kg/667m
2, control group is 598kg/667m
2Stimulation ratio is 15.9%.
Soybean
Kind: east farming 42.Sowing date: on May 8th, 2011.Harvest date: treatment group: on September 25th, 2011; Control group: on September 30th, 2011.Test place: Yong Fa village, Suihua City Golconda town, Heilongjiang Province; Soil characteristic: continuous cropping 3 years.
Conclusion: the treatment group soybean emerged May 18, and it is neat to emerge, and growth of seedling is healthy and strong, and the thick leaf color depth of stem; In 3 days in advance flowering period, bear pods after spending closely; Full seed.Vegetative period, disease was few.Treatment group was gathered Zao 5 days than control group.The output investigation, treatment group output is 257kg/667m
2, control group is 222kg/667m
2Stimulation ratio is 15.8%.
Paddy rice
Kind: east farming 425.Grow seedlings the date: on April 9th, 2011.The field planting date: on May 27th, 2011.Harvest date: on September 30th, 2011.Test place: Yong Fa village, Suihua City Golconda town, Heilongjiang Province; Soil characteristic: continuous cropping 3 years.
Conclusion: after the field planting of treatment group paddy rice, slow seedling is fast, well developed root system, tiller many; Robust plant, the leaf look dark green, and disease and pest is few; The output investigation, treatment group output is 601kg/667m
2, control group is 558kg/667m
2Stimulation ratio is 7.7%.
Capsicum
Kind: scape green pepper No. 1.Grow seedlings the date: on March 9th, 2011.The field planting date: on May 21st, 2011.Begin the date of gathering: on July 27th, 2011.Test place: Tong De village, the concentric township of Heilongjiang Province's Shuangcheng City; Soil characteristic: continuous cropping 7 years.
Conclusion: after the field planting of treatment group capsicum, slow seedling is fast, and seedling changes growth conditions fast over to; Dark green leaf color, and stem is slightly thick than control group; The individual plant area is big; First florescence comparison was according to early 2 days; Fruit begins to pluck treatment group than control group Zao 4 days; Fruit-setting rate is high; Fruiting period is long; The treatment group fruit marketability is good, and fruit color is dark green, bright in color, profile are neat; The treatment group disease is few; Finish late 10 days of harvesting time than control group.The output investigation, treatment group output is 2882kg/667m
2, control group is 2331kg/667m
2Stimulation ratio is 23.6%.
Yam
Kind: restrain new 12.Sowing date: on April 30th, 2011.Harvesting time: on September 27th, 2011.Test place: Heilongjiang Province Hailun City truth potato planting cooperative society; Soil issues: the first crop poisoning is serious.
Conclusion: the treatment group potato seedling is emerged soon, and plant is healthy and strong, and blade is big, dark green leaf color, and stem is slightly thick than control group; First florescence comparison was according to early 2 days; Vegetative period, disease was few; Control group receives first crop poisoning influence, a little less than the seedling, wilts, and blade is yellow; Treatment group knot yam is many and big; Fruit marketability is good, and profile is big and neat; Treatment group and control group are gathered simultaneously.The output investigation, treatment group output is 2245kg/667m
2, control group is 1783kg/667m
2Stimulation ratio is 25.9%.
Watermelon
Kind: Jing Fengbao.Sowing date: on May 15th, 2011.Harvesting time: on August 12nd, 2011.Test place: Tong De village, the concentric township of Heilongjiang Province's Shuangcheng City; Soil issues: continuous cropping 2 years.
Conclusion: the treatment group watermelon, growth potential is strong, and disease resistance is strong, and percentage of fertile fruit is high; Dark green leaf color, stem are slightly thick than control group; First florescence comparison was according to early 3 days; The treatment group fruit marketability is good, and bright in color, profile are neat, and pol is high.The output investigation, treatment group output is 5448kg/667m
2, control group is 4305kg/667m
2Stimulation ratio is 26.6%.
Tomato
Kind: east farming 708.Grow seedlings the date: on January 5th, 2012, field planting date: on April 15th, 2012.Begin collection period: on June 23rd, 2012.Test place: the bright village of Harbin City, Heilongjiang Province city Xiangfang District; Soil characteristic: booth continuous cropping 7 years.
Conclusion: after the field planting of treatment group tomato, slow seedling is fast, and seedling changes growth conditions fast over to; Dark green leaf color, stem are slightly thick than control group; The individual plant area is big; First florescence comparison was according to early 3 days; Fruit begins to pluck treatment group than control group Zao 4 days; Fruit-setting rate is high, fruit is big; The treatment group fruit marketability is good, and fruit color is bright-coloured, bright in color, profile are neat; The treatment group disease is few.The output investigation, treatment group output is 7552kg/667m
2, control group is 6281kg/667m
2Stimulation ratio is 20.2%.
The probiotic type soil conditioner product that the present invention obtains not only can be applied to nurse one's health soil, also can be used as corps leaf surface simultaneously and sprays application such as defending and fighting against diseases, microbial fermentation startup agent, the deodorization of livestock and poultry house disease resistance.
The present invention is not limited only to the scope of said embodiment, and all foundations working method of the present invention suitably develops, and resulting working method all belongs within protection scope of the present invention.
Claims (6)
1. the working method of a probiotic type soil conditioner is characterized in that, comprising:
(1) produces preparation: bacillus megaterium, subtilis, Bacillus licheniformis are inserted seed culture medium respectively cultivate with the probiotic bacterium seed liquor; Obtain bacillus megaterium seed liquor, subtilis seed liquor and Bacillus licheniformis seed liquor; Wherein the incubation time of bacillus megaterium is 8-14h, and the incubation time of subtilis and Bacillus licheniformis is 6-12h;
(2) produce preparation and sterilization: will produce with in the substratum adding fermentor tank, and sterilize 30-45 minute, and be cooled to 30-35 ℃ for 115-121 ℃ with substratum;
(3) inoculation and fermentation: according to using the ratio access fermentor tank of the volume ratio of substratum as 2-5% with said production, at 29-33 ℃, logical oxygen agitation condition is cultivated 10-18h down with said bacillus megaterium seed liquor; Then leavening temperature is adjusted to 33-37 ℃; Insert said subtilis seed liquor and Bacillus licheniformis seed liquor; Add simultaneously with said production and use the substratum TV, continue to cultivate 7-16h, obtain said probiotic type soil conditioner than concentrated sterilising medium as 4-10%.
2. the working method of probiotic type soil conditioner as claimed in claim 1 is characterized in that: said production is following with the prescription of substratum, and every liter of production is with containing in the substratum: sucrose 5-15g; Lactose 5-12g, SANMALT-S 1-5g, Zulkovsky starch 1-5g; Peptone 5-15g, yeast extract paste 2-9g, urea 1-5g; NaCl 3-8g, K
2SO
40.4-1.0g, K
2HPO
40.5-1.5g, MgSO
47H
2O 2-3.5g, CaCl
20.8-1.5g, CaCO
34-6.5g surplus is a water; PH7.5-8.2.
3. the working method of probiotic type soil conditioner as claimed in claim 1 is characterized in that: the said subtilis seed liquor of access and said Bacillus licheniformis seed liquor volume ratio are 1:0.25-4.
4. the working method of probiotic type soil conditioner as claimed in claim 1 is characterized in that: said subtilis seed liquor is used culture volume than being 4-8% with the access total amount of said Bacillus licheniformis seed liquor with said production.
5. the working method of probiotic type soil conditioner as claimed in claim 1 is characterized in that: the said concentrated bactericidal nurishing base of adding consists of: every liter of concentrated sterilising medium contains: sucrose 30-50g, lactose 30-40g; SANMALT-S 5-15g; Zulkovsky starch 5-15g, peptone 25-35g, yeast extract paste 20-30g; NaCl5-10g, K
2HPO
40.8-1.8g, MgSO
47H
2O 2-4g, CaCO
35-15g, surplus is a water; PH7.5-8.2.
6. like the probiotic type soil conditioner of the working method of each described probiotic type soil conditioner of claim 1 ~ 5 preparation; It is characterized in that: living bacteria count adds up to hundred million/milliliter of 50-100 in the said probiotic type soil conditioner; The gemma rate is at 90-100%, and the living bacteria count of every kind of bacterium is greater than 200,000,000/milliliter.
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