CN105237280A - Microorganism in-problem soil ecological restoration preparation, preparation method and application thereof - Google Patents
Microorganism in-problem soil ecological restoration preparation, preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a microorganism in-problem soil ecological restoration preparation, which is prepared through a multi-genus multi-bacterial strain synthetic acclimatization cultivation technology, wherein the preparation is prepared with a mixed base material containing dehydrated cow dung and the like as raw materials with bacterial strains in multi-genera through solid fermentation and drying and shaping. The invention aims to solve the key point of imbalance of micro-ecological environment of various in-problem soil and also aims to solve the problem of treatment of the cow dung from pastures, and discloses a preparation method of the ecological restoration preparation, which is prepared with the cow dung as main base material and aims to the in-problem soil, through the multi-genus multi-bacterial strain synthetic acclimatization cultivation technology, thereby enlarging utilization scope of the cow dung and finding a new approach of solving the problem of environment pollution due to the cow dung. The invention, at the same time, provides the ecological restoration preparation for the in-problem soil to achieve sustainable utilization and zero secondary pollution.
Description
Technical field
The present invention relates to livestock industry and field of agricultural cultivation, especially livestock industry and field of agricultural cultivation are combined to obtain a kind of problematic soils restoration of the ecosystem preparation, particularly, relate to a kind of microorganism problem soil ecology preparation for repairing and its preparation method and application.
Background technology
Under national policy is supported, under the surging great number revenue impact brought of beef price, domestic cattle-raising has only used the time less than 30 years, is just mainly from one the household sideline production that plant husbandry provides animal power, develops into a commodity beef cattle in large scale and dairy.The breeding stock of beef cattle reaches about 3,000 ten thousand, and the breeding stock of milk cow reaches about 1,400 ten thousand.General ox year Fecal wastes more than 12 tons, so the annual production of cow dung at least can reach more than 52,800 ten thousand tons." resource " of huge amount like this, at present except a small amount of cow dung is for the production of biogas, as except fuel power generation function, fermented manure, starting material, cow dung brings great puzzlement to milk cow and beef cattle feeding field.Due to the organic matter of cow dung and nutrient content minimum in various domestic animal, be of close texture, moisture more, decompose slow, thermal value is low, and compost is ripe slow, and cow dung can not be allowed thoroughly to become thoroughly decomposed, and easily " Secondary Fermentation " occurs after applying farmland, causes and burns the phenomenon that root burns seedling.Various reasons like this makes cow dung fully do not utilized, and causes cow dung in the accumulation of feedlot, and form foul smell, external leakage pollution environment, affects the quality of life of nearby residents greatly.Therefore, the recycling scope developing cow dung is the difficult problem that in livestock industry, continue for for many years.
Meanwhile, China's cultivated area (the second national land investigation result that the end of the year 2013 announced, national cultivated area totally 20.3 hundred million mu) the row world the 4th, but per capita cultivated land only 1.4 mu, be less than the half of world's per capita area of cultivated farmland, after coming the 126th of the world.Although; obviously treasure these arable lands of protection and seem particularly important; but " soil pollution and the HUMAN HEALTH " of Chinese Ministry of Environmental Protection's distribution in 2013 claims; the whole nation only just reaches 300,000,000 mu by the cultivated area of heavy metal contamination; mainly ploughing in distribution province in 18, the whole nation, has 13 to there is heavy metal-polluted soil excessive problem.In addition live discharge pollution, the destruction problem that the chemicals such as chemical fertilizer, agricultural chemicals, weedicide cause soil, the problem on obstacle of being punished for being related to that the edatope also having plant growing itself to cause changes, especially various pollution problem has caused current arable land to exceed to guard against red line and has threatened the essence safety in arable land.Analyze many problematic soils and have a common feature, be exactly in soil microecological balance destroyed, harmful microorganism, harmful insect class account for competitive edge, independent breeding expands, cause Soil structure to change, metal ion is convenient to enrichment, and disease and pest is easy to the plantation obstacle of generation etc.The solution of these problems needs the ecological soil reparation not having secondary pollution, and soil amelioration techniques solves.
Have the agent of heavy metal-polluted soil fixation in situ complex repairation in the market, repair pollution of herbicide soil microorganisms preparation, repair polycyclic aromatic hydrocarbon pollution microbial preparation, microbe soil restoration agent belongs to microbial fertilizer, combined soil renovation agent etc. containing microbial bacteria, these microbe soil restoration agents are all to spread cultivation specific microbial strains with microbiological culture media, or other fertilizer of remix, the still interpolation chemical composition had and make soil remediation formulation.Further, these products very easily cause secondary pollution or soil Dependence Problem in actual application, therefore also not as people's will.
Summary of the invention
The present invention is conceived to the unbalanced main points of Tiny ecosystem of various problematic soils, be conceived to the cow dung process problem in pasture simultaneously, seek a kind of with cow dung be main matrix, for problematic soils basic problem, utilize many Pseudomonas multi-cultur es to synthesize the preparation method of domestication culture technique production restoration of the ecosystem preparation, to expand the scope that cow dung utilizes, a new outlet is opened up for solving cow dung problem of environmental pollution, meanwhile, for problematic soils provides sustainable, there is no the restoration of the ecosystem preparation of secondary pollution.Further, present invention also offers a kind of microorganism problem soil ecology preparation for repairing by described preparation method.
Technical scheme of the present invention comprises a kind of microorganism problem soil ecology preparation for repairing, adopt many Pseudomonas multi-cultur es synthesis domestication culture technique, to comprise the hybrid substrate of dehydration cow dung for raw material, after carrying out solid fermentation by many Pseudomonas bacterial classification, drying and moulding obtains;
Wherein, the component of described many Pseudomonas bacterial classification and weight proportion thereof comprise:
In one embodiment of the invention, the water content of described hybrid substrate is 30-40wt%, Ph is 7.0-9.0, and its component and weight proportion thereof comprise:
Further, described auxiliary material is selected from any one or a few in diatomite, opoka, rock ash, spongolite; Described stalk be selected from rice straw, maize straw, wheat stalk and other paddy wheat class stalk any one or a few.
In one embodiment, described microorganism problem soil ecology preparation for repairing is brown powdered granule.
Technical scheme of the present invention also comprises a kind of preparation method of microorganism problem soil ecology preparation for repairing described above, and described preparation method comprises:
Step 1: many Pseudomonas bacterial classification is cultivated in synthesis domestication
(1) preparation of Microbe synthesis thalline suspension liquid
Trichoderma, Aspergillus candidus, streptomycete, subtilis, Candida humicola, blown-ball Azotobacter bacterial classification are added in synthesis domestication substratum, under the stirring velocity, 30-60 DEG C condition of 150-200rpm, carry out synthesis domestication cultivate 5-7 days, obtain Microbe synthesis microbial inoculum, it is mixed with salt solution and protective material, obtains Microbe synthesis thalline suspension liquid;
(2) Rhodopseudomonas palustris spreads cultivation the preparation of liquid
Added by Rhodopseudomonas palustris in basic medium, under natural lighting, 28-35 DEG C condition, spread cultivation 8-12 days, until in light red to scarlet, obtain Rhodopseudomonas palustris and to spread cultivation liquid;
Step 2: preparation hybrid substrate
Process cow dung makes its water content be down to 50-60wt%, obtains cow dung of dewatering; Described dehydration cow dung, pig manure, the stalk husk of pulverizing, auxiliary material are fully mixed, regulates water content to 30-40wt%, pH to 7.0-9.0, obtain described hybrid substrate;
Step 3: prepare microorganism problem soil ecology preparation for repairing standby
The liquid that Microbe synthesis thalline suspension liquid and Rhodopseudomonas palustris spread cultivation adds in described hybrid substrate, stir, and aerobic fermentation is carried out under the stirring velocity of 2-30rpm, wherein, earlier fermentation temperature controls be less than 65 DEG C, and fermentation latter temperature controls at 30-50 DEG C, fermentation 10-15 days, until fermentation material be light gray to Dark grey, and with fermenting aroma, terminate fermentation; Further fermentation material is dried to water content lower than 10wt% under lower than 50 DEG C of conditions, obtains described microorganism problem soil ecology preparation for repairing.
Further, described preparation method also comprises the preparation of each bacterial classification, wherein, by the Trichoderma (Trichodermasp. bought from Chinese agriculture Microbiological Culture Collection administrative center, ACCC31526), Aspergillus candidus (Aspergiluscandidus, ACCC30347), streptomycete (streptomycesStreptomycessp., ACCC41239), subtilis (Bacilussubtilis, ACCC01185), Candida humicola (Candidahumicola, ACCC20112), blown-ball Azotobacter (Azotobacterchroococcum, ACCC10003), Rhodopseudomonas palustris (Rhodopseudomonaspalustris, ACCC10649) carry out respectively cultivating each bacterial classification obtained for subsequent step.
Particularly, Trichoderma and Candida humicola access potato dextrose broth (are got peeled potatoes 200 grams, be cut into small pieces, add water 1000 milliliters and boil 30 minutes, elimination potato ball, filtrate is complemented to 1000 milliliters, add glucose 20 grams) Erlenmeyer flask in, shaking culture 48-96 hour under 28-32 DEG C of condition, takes out after substratum muddiness, then puts into 2-4 DEG C of Refrigerator store stand-by;
Aspergillus candidus is transferred in the Erlenmeyer flask of malt juice liquid medium, 23-28 DEG C, cultivate 4-8 days under 180-220rpm oscillating condition, take out after substratum muddiness, then put into 2-4 DEG C of Refrigerator store stand-by;
Streptomycete is transferred into ISP2 substratum (yeast extract paste 4.0g/L, wort 10.0g/L, glucose 4.0g/L, pH is 7.0) Erlenmeyer flask in, 26-32 DEG C, cultivate 48-72 hour under 180-220rpm oscillating condition, take out after substratum muddiness, then put into 2-4 DEG C of Refrigerator store stand-by;
Transferred by subtilis extractum carnis 1%, peptone 0.5%, dipotassium hydrogen phosphate 0.1%, pectin 0.5%, sodium-chlor 0.3%, PH is in the liquid nutrient medium Erlenmeyer flask of 7.5,26-32 DEG C, cultivate 48-72 hour under 180-220rpm oscillating condition, take out after substratum muddiness, then put into 2-4 DEG C of Refrigerator store stand-by;
Blown-ball Azotobacter is transferred into glucose 20g/L, K
2hPO
41.0g/L, MgSO
40.2g/L, CaCO
30.5g/L, FeCl
30.25g/L, NaMoO
40.5g/L and with distilled water preparation pH be the liquid nutrient medium of 7.3 ~ 7.6, it is placed in the Erlenmeyer flask with granulated glass sphere, carries out shaking table cultivation, rotating speed be under 180-220rpm, 28-35 DEG C of condition cultivate 32 ~ 48h;
Rhodopseudomonas palustris is transferred into CH
3cOONa3.0g/L, NaHCO
35.0g/L, (NH
4)
2s0
41.0g/L, peptone 5.0g/L, yeast extract paste 1.0g/L, MgS0
40.5g/L, NaCl1.0g/L, KH
2p0
41.0g/L, K
2hPO
40.5g/L, CaCl
20.05g/L, CaCO
320g/L, pH are in the liquid nutrient medium of 7.0-7.5, leave standstill illumination (light intensity 2000lx), cultivate 4-7 days for 28-35 DEG C.
Further, described synthesis domestication substratum is by by wort 1-3 weight part, murphy juice 0.5-2.0 weight part, yeast extract paste 0.3-0.6 weight part, brown sugar 2-5 weight part, protective material 2-5 weight part, MgSO
40.01 weight part, CaCO
30.02 weight part, FeCl
30.01 weight part, NaCl0.01 weight part, water 80-98 weight part mix, and regulate pH to 6.8-7.5 to obtain.
Described protective material be selected from sucrose, glucose, honey, molasses any one or a few.
Described basic medium is CH
3cOONa3.0g/L, NaHCO
35.0g/L, (NH
4)
2s0
41.0g/L, peptone 5.0g/L, yeast extract paste 1.0g/L, MgS0
40.5g/L, NaCl1.0g/L, KH
2p0
41.5g/L, K
2hPO
40.5g/L, CaCl
20.05g/L, CaCO
320g/L, pH are 7.0-7.5, and carbon-nitrogen ratio is the liquid nutrient medium of 1.5.
In a preferred embodiment of the invention, described preparation method specifically comprises the steps:
Step 1: many Pseudomonas bacterial classification is cultivated in synthesis domestication
(1) preparation of Microbe synthesis thalline suspension liquid
According to Trichoderma 10-25 part, Aspergillus candidus 10-25 part, streptomycete 20-30 part, subtilis 15-25 part, Candida humicola 10-25 part, the weight proportion of blown-ball Azotobacter 10-25 part is joined and is got each bacterial classification, and with the weight proportion of 1:80-120, described each bacterial classification is added in mixed-culture medium, put into fermentor tank, 5-7 days is cultivated under the condition of the stirring velocity of 150-200rpm and 30-60 DEG C, until present yellow to filbert, obtain Microbe synthesis microbial inoculum, by the 30wt% salt solution of itself and 20-40 weight part, the 20wt% Industry Waste molasses mixing of 20-40 weight part, obtain Microbe synthesis thalline suspension liquid, wherein, described mixed-culture medium is by by wort 1-3 weight part, murphy juice 0.5-2.0 weight part, yeast extract paste 0.3-0.6 weight part, brown sugar 2-5 weight part, honey 2-5 weight part, MgSO
40.01 weight part, CaCO
30.02 weight part, FeCl
30.01 weight part, NaCl0.01 weight part, water 80-98 weight part mix, and regulate pH to 6.8-7.5 to obtain,
(2) Rhodopseudomonas palustris spreads cultivation the preparation of liquid
Added by Rhodopseudomonas palustris in basic medium, under natural lighting, 28-35 DEG C condition, spread cultivation 8-12 days, until in light red to scarlet, obtain Rhodopseudomonas palustris and to spread cultivation liquid; Wherein, described basic medium is CH
3cOONa3.0g/L, NaHCO
35.0g/L, (NH
4)
2s0
41.0g/L, peptone 5.0g/L, yeast extract paste 1.0g/L, MgS0
40.5g/L, NaCl1.0g/L, KH
2p0
41.5g/L, K
2hPO
40.5g/L, CaCl
20.05g/L, CaCO
320g/L, pH are 7.0-7.5, and carbon-nitrogen ratio is the liquid nutrient medium of 1.5;
Step 2: preparation hybrid substrate
Will containing organic 11.0-14.5 weight part, nitrogen 0.30-0.45 weight part, phosphorus (P
2o
5) 0.15-0.25 weight part, potassium (K
2o) 0.10-0.15 weight part and water content are that the cow dung of 80.0-83.3wr% processes, and are down to 50-60wt% to cow dung water content, obtain cow dung of dewatering; Described dehydration cow dung 50-70 weight part, pig manure 10-20 weight part, the stalk husk 10-20 weight part of pulverizing, auxiliary material 0.5-3.5 weight part are fully mixed, regulates water content to 30-40wt%, pH to 7.0-9.0, obtain described hybrid substrate;
Step 3: prepare microorganism problem soil ecology preparation for repairing standby
Spread cultivation Microbe synthesis thalline suspension liquid 5-15 weight part and Rhodopseudomonas palustris liquid 1-10 weight part, add in described hybrid substrate, stir, under the stirring velocity of 2-30rpm, carry out aerobic fermentation, wherein, earlier fermentation, the temperature namely in 5 days controls be less than 65 DEG C, and fermentation latter temperature controls at 30-50 DEG C, fermentation 10-15 days, until fermentation material be light gray to Dark grey, and with fermenting aroma, terminate fermentation; Fermentation material is dried to water content lower than 10wt% under lower than 50 DEG C of conditions, and obtain described microorganism problem soil ecology preparation for repairing, wherein, total viable count is greater than 5*10
8cfu/g, and harmless treat index reaches the standard-required of GB GB20287.
Technical scheme of the present invention also comprises a kind of application of microorganism problem soil ecology preparation for repairing described above, comprises the reparation for the reparation of Soil-sickness Problem soil and house site, agriculture chemical pollution problem soil.
Preferably, the usage quantity of described microorganism problem soil ecology preparation for repairing when embody rule is 1 ~ 3kg/ mu.
Particularly:
The reparation of Soil-sickness Problem soil: after front stubble results, every mu of ground evenly spills into 1-5kg, ploughs; During rear stubble whole ground, every mu of ground evenly spills into 0.5-3.0kg again;
The problematic soils reparations such as house site, agriculture chemical pollution use:
Every mu of ground evenly spills into 3-8kg, ploughs deeply ground, and after 1-3 month, second time is every per muly evenly spills into 1-5kg, ploughs, and after 1-3 month, third time is every per muly evenly spills into 1-3kg, prepares plantation wholely.
Advantage of the present invention is: the low cost being main matrix with pasture cow dung, short compared with the time of cow dung Natural compost, tame and docile the basic bacterial classification of many Pseudomonas bacterial classification as microorganism problem soil ecology preparation for repairing closing and cultivate and there is different metabolic and synthesize ecologic active product, utilize many Pseudomonas bacterial classification to act synergistically, efficiently prepare problematic soils restoration of the ecosystem preparation.
Further, the bacterial classification of many Pseudomonas (actinomyces, mould Pseudomonas, bacterium genus, photosynthetic Pseudomonas etc.) that problematic soils restoration of the ecosystem preparation of the present invention adopts, the synergy of these many Pseudomonas bacterial classifications, enable this problematic soils restoration of the ecosystem preparation to soil various problems, as restoration of the ecosystem is carried out in the pollution problem such as problem, agriculture chemical of restoreing abandoned land to cultivation of problem on obstacle of being punished for being related to, house site, more can not cause second time pollution problem, make soil progressively recover self eubiosis and lasting repair ability.
Terminological interpretation:
Problematic soils: problematic soils comprises the soil of heavy metals exceeding standard problem, is destroyed the soil of physicochemical structure problem by chemicals such as chemical fertilizer, agricultural chemicals, weedicides, the soil etc. of the problem on obstacle of being punished for being related to that the edatope also having plant farming itself to cause changes.
Many Pseudomonas multi-cultur es synthesis domestication culture technique: in order to many Pseudomonas multi-cultur es can be enable to play synergy, and create the technology of domestication's envrionment conditions.
Embodiment
The invention provides a kind of microorganism problem soil ecology preparation for repairing, be adopt many Pseudomonas multi-cultur es synthesis domestication culture technique, to comprise the hybrid substrate of dehydration cow dung for raw material, after carrying out solid fermentation by many Pseudomonas bacterial classification, drying and moulding obtains;
Component and the weight proportion thereof of described many Pseudomonas bacterial classification comprise:
In one embodiment of the invention, the water content of described hybrid substrate is 30-40wt%, Ph is 7.0-9.0, and its component and weight proportion thereof comprise:
Further, described auxiliary material is selected from any one or a few in diatomite, opoka, rock ash, spongolite; Described stalk be selected from rice straw, maize straw, wheat stalk and other paddy wheat class stalk any one or a few.
In one embodiment, described microorganism problem soil ecology preparation for repairing is brown powdered granule.
Present invention also offers a kind of preparation method of microorganism problem soil ecology preparation for repairing described above, described preparation method comprises:
Step 1: many Pseudomonas bacterial classification is cultivated in synthesis domestication
(1) preparation of Microbe synthesis thalline suspension liquid
Trichoderma, Aspergillus candidus, streptomycete, subtilis, Candida humicola, blown-ball Azotobacter bacterial classification are added in synthesis domestication substratum, under the stirring velocity, 30-60 DEG C condition of 150-200rpm, carry out synthesis domestication cultivate 5-7 days, obtain Microbe synthesis microbial inoculum, it is mixed with salt solution and protective material, obtains Microbe synthesis thalline suspension liquid;
(2) Rhodopseudomonas palustris spreads cultivation the preparation of liquid
Added by Rhodopseudomonas palustris in basic medium, under natural lighting, 28-35 DEG C condition, spread cultivation 8-12 days, until in light red to scarlet, obtain Rhodopseudomonas palustris and to spread cultivation liquid;
Step 2: preparation hybrid substrate
Process cow dung makes its water content be down to 50-60wt%, obtains cow dung of dewatering; Described dehydration cow dung, pig manure, the stalk husk of pulverizing, auxiliary material are fully mixed, regulates water content to 30-40wt%, pH to 7.0-9.0, obtain described hybrid substrate;
Step 3: prepare microorganism problem soil ecology preparation for repairing standby
The liquid that Microbe synthesis thalline suspension liquid and Rhodopseudomonas palustris spread cultivation adds in described hybrid substrate, stir, and aerobic fermentation is carried out under the stirring velocity of 2-30rpm, wherein, earlier fermentation temperature controls be less than 65 DEG C, and fermentation latter temperature controls at 30-50 DEG C, fermentation 10-15 days, until fermentation material be light gray to Dark grey, and with fermenting aroma, terminate fermentation; Further fermentation material is dried to water content lower than 10wt% under lower than 50 DEG C of conditions, obtains described microorganism problem soil ecology preparation for repairing.
Further, described preparation method also comprises the preparation of each bacterial classification, wherein, by the Trichoderma (Trichodermasp. bought from Chinese agriculture Microbiological Culture Collection administrative center, ACCC31526), Aspergillus candidus (Aspergiluscandidus, ACCC30347), streptomycete (streptomycesStreptomycessp., ACCC41239), subtilis (Bacilussubtilis, ACCC01185), Candida humicola (Candidahumicola, ACCC20112), blown-ball Azotobacter (Azotobacterchroococcum, ACCC10003), Rhodopseudomonas palustris (Rhodopseudomonaspalustris, ACCC10649) carry out respectively cultivating each bacterial classification obtained for subsequent step.
Particularly, Trichoderma and Candida humicola access potato dextrose broth (are got peeled potatoes 200 grams, be cut into small pieces, add water 1000 milliliters and boil 30 minutes, elimination potato ball, filtrate is complemented to 1000 milliliters, add glucose 20 grams) Erlenmeyer flask in, shaking culture 48-96 hour under 28-32 DEG C of condition, takes out after substratum muddiness, then puts into 2-4 DEG C of Refrigerator store stand-by;
Aspergillus candidus is transferred in the Erlenmeyer flask of malt juice liquid medium, 23-28 DEG C, cultivate 4-8 days under 180-220rpm oscillating condition, take out after substratum muddiness, then put into 2-4 DEG C of Refrigerator store stand-by;
Streptomycete is transferred into ISP2 substratum (yeast extract paste 4.0g/L, wort 10.0g/L, glucose 4.0g/L, pH is 7.0) Erlenmeyer flask in, 26-32 DEG C, cultivate 48-72 hour under 180-220rpm oscillating condition, take out after substratum muddiness, then put into 2-4 DEG C of Refrigerator store stand-by;
Transferred by subtilis extractum carnis 1%, peptone 0.5%, dipotassium hydrogen phosphate 0.1%, pectin 0.5%, sodium-chlor 0.3%, PH is in the liquid nutrient medium Erlenmeyer flask of 7.5,26-32 DEG C, cultivate 48-72 hour under 180-220rpm oscillating condition, take out after substratum muddiness, then put into 2-4 DEG C of Refrigerator store stand-by;
Blown-ball Azotobacter is transferred into glucose 20g/L, K
2hPO
41.0g/L, MgSO
40.2g/L, CaCO
30.5g/L, FeCl
30.25g/L, NaMoO
40.5g/L and with distilled water preparation pH be the liquid nutrient medium of 7.3 ~ 7.6, it is placed in the Erlenmeyer flask with granulated glass sphere, carries out shaking table cultivation, rotating speed be under 180-220rpm, 28-35 DEG C of condition cultivate 32 ~ 48h;
Rhodopseudomonas palustris is transferred into CH
3cOONa3.0g/L, NaHCO
35.0g/L, (NH
4)
2s0
41.0g/L, peptone 5.0g/L, yeast extract paste 1.0g/L, MgS0
40.5g/L, NaCl1.0g/L, KH
2p0
41.0g/L, K
2hPO
40.5g/L, CaCl
20.05g/L, CaCO
320g/L, pH are in the liquid nutrient medium of 7.0-7.5, leave standstill illumination (light intensity 2000lx), cultivate 4-7 days for 28-35 DEG C.
Further, described synthesis domestication substratum is by by wort 1-3 weight part, murphy juice 0.5-2.0 weight part, yeast extract paste 0.3-0.6 weight part, brown sugar 2-5 weight part, protective material 2-5 weight part, MgSO
40.01 weight part, CaCO
30.02 weight part, FeCl
30.01 weight part, NaCl0.01 weight part, water 80-98 weight part mix, and regulate pH to 6.8-7.5 to obtain.
Described protective material be selected from sucrose, glucose, honey, molasses any one or a few.
Described basic medium is CH
3cOONa3.0g/L, NaHCO
35.0g/L, (NH
4)
2s0
41.0g/L, peptone 5.0g/L, yeast extract paste 1.0g/L, MgS0
40.5g/L, NaCl1.0g/L, KH
2p0
41.5g/L, K
2hPO
40.5g/L, CaCl
20.05g/L, CaCO
320g/L, pH are 7.0-7.5, and carbon-nitrogen ratio is the liquid nutrient medium of 1.5.
In a preferred embodiment of the invention, described preparation method specifically comprises the steps:
Step 1: many Pseudomonas bacterial classification is cultivated in synthesis domestication
(1) preparation of Microbe synthesis thalline suspension liquid
According to Trichoderma 10-25 part, Aspergillus candidus 10-25 part, streptomycete 20-30 part, subtilis 15-25 part, Candida humicola 10-25 part, the weight proportion of blown-ball Azotobacter 10-25 part is joined and is got each bacterial classification, and with the weight proportion of 1:80-120, described each bacterial classification is added in mixed-culture medium, put into fermentor tank, 5-7 days is cultivated under the condition of the stirring velocity of 150-200rpm and 30-60 DEG C, until present yellow to filbert, obtain Microbe synthesis microbial inoculum, by the 30wt% salt solution of itself and 20-40 weight part, the 20wt% Industry Waste molasses mixing of 20-40 weight part, obtain Microbe synthesis thalline suspension liquid, wherein, described mixed-culture medium is by by wort 1-3 weight part, murphy juice 0.5-2.0 weight part, yeast extract paste 0.3-0.6 weight part, brown sugar 2-5 weight part, honey 2-5 weight part, MgSO
40.01 weight part, CaCO
30.02 weight part, FeCl
30.01 weight part, NaCl0.01 weight part, water 80-98 weight part mix, and regulate pH to 6.8-7.5 to obtain,
(2) Rhodopseudomonas palustris spreads cultivation the preparation of liquid
Added by Rhodopseudomonas palustris in basic medium, under natural lighting, 28-35 DEG C condition, spread cultivation 8-12 days, until in light red to scarlet, obtain Rhodopseudomonas palustris and to spread cultivation liquid; Wherein, described basic medium is CH
3cOONa3.0g/L, NaHCO
35.0g/L, (NH
4)
2s0
41.0g/L, peptone 5.0g/L, yeast extract paste 1.0g/L, MgS0
40.5g/L, NaCl1.0g/L, KH
2p0
41.5g/L, K
2hPO
40.5g/L, CaCl
20.05g/L, CaCO
320g/L, pH are 7.0-7.5, and carbon-nitrogen ratio is the liquid nutrient medium of 1.5;
Step 2: preparation hybrid substrate
Will containing organic 11.0-14.5 weight part, nitrogen 0.30-0.45 weight part, phosphorus (P
2o
5) 0.15-0.25 weight part, potassium (K
2o) 0.10-0.15 weight part and water content are that the cow dung of 80.0-83.3wr% processes, and are down to 50-60wt% to cow dung water content, obtain cow dung of dewatering; Described dehydration cow dung 50-70 weight part, pig manure 10-20 weight part, the stalk husk 10-20 weight part of pulverizing, auxiliary material 0.5-3.5 weight part are fully mixed, regulates water content to 30-40wt%, pH to 7.0-9.0, obtain described hybrid substrate;
Step 3: prepare microorganism problem soil ecology preparation for repairing standby
Spread cultivation Microbe synthesis thalline suspension liquid 5-15 weight part and Rhodopseudomonas palustris liquid 1-10 weight part, add in described hybrid substrate, stir, under the stirring velocity of 2-30rpm, carry out aerobic fermentation, wherein, earlier fermentation, the temperature namely in 5 days controls be less than 65 DEG C, and fermentation latter temperature controls at 30-50 DEG C, fermentation 10-15 days, until fermentation material be light gray to Dark grey, and with fermenting aroma, terminate fermentation; Fermentation material is dried to water content lower than 10wt% under lower than 50 DEG C of conditions, and obtain described microorganism problem soil ecology preparation for repairing, wherein, total viable count is greater than 5*10
8cfu/g, and harmless treat index reaches the standard-required of GB GB20287.
Present invention also offers a kind of application of microorganism problem soil ecology preparation for repairing described above, comprise the reparation for the reparation of Soil-sickness Problem soil and house site, agriculture chemical pollution problem soil.
Preferably, the usage quantity of described microorganism problem soil ecology preparation for repairing when embody rule is 1 ~ 3kg/ mu.
Particularly:
The reparation of Soil-sickness Problem soil: after front stubble results, every mu of ground evenly spills into 1-5kg, ploughs; During rear stubble whole ground, every mu of ground evenly spills into 0.5-3.0kg again;
The problematic soils reparations such as house site, agriculture chemical pollution use:
Every mu of ground evenly spills into 3-8kg, ploughs deeply ground, and after 1-3 month, second time is every per muly evenly spills into 1-5kg, ploughs, and after 1-3 month, third time is every per muly evenly spills into 1-3kg, prepares plantation wholely.
Below by following examples and Application Example, technical scheme of the present invention and beneficial effect are further explained and are illustrated.
Embodiment
A kind of preparation method of microorganism problem soil ecology preparation for repairing
Step 1: the preparation of each bacterial classification
By the Trichoderma (Trichodermasp. bought from Chinese agriculture Microbiological Culture Collection administrative center, ACCC31526), Aspergillus candidus (Aspergiluscandidus, ACCC30347), streptomycete (streptomycesStreptomycessp., ACCC41239), subtilis (Bacilussubtilis, ACCC01185), Candida humicola (Candidahumicola, ACCC20112), blown-ball Azotobacter (Azotobacterchroococcum, ACCC10003), Rhodopseudomonas palustris (Rhodopseudomonaspalustris, ACCC10649) carry out respectively cultivating each bacterial classification obtained for subsequent step, particularly:
Trichoderma and Candida humicola access potato dextrose broth (are got peeled potatoes 200 grams, be cut into small pieces, add water 1000 milliliters and boil 30 minutes, elimination potato ball, filtrate is complemented to 1000 milliliters, adds glucose 20 grams) Erlenmeyer flask in, shaking culture 72 hours under 30 DEG C of conditions, take out after substratum muddiness, then put into 2 DEG C of Refrigerator stores stand-by;
Aspergillus candidus is transferred in the Erlenmeyer flask of malt juice liquid medium, 25 DEG C, cultivate 7 days under 180rpm oscillating condition, take out after substratum muddiness, then put into 2 DEG C of Refrigerator stores stand-by;
Streptomycete is transferred into ISP2 substratum (yeast extract paste 4.0g/L, wort 10.0g/L, glucose 4.0g/L, pH is 7.0) Erlenmeyer flask in, 30 DEG C, cultivate 72 hours under 180rpm oscillating condition, take out after substratum muddiness, then put into 2 DEG C of Refrigerator stores stand-by;
Transferred by subtilis extractum carnis 1%, peptone 0.5%, dipotassium hydrogen phosphate 0.1%, pectin 0.5%, sodium-chlor 0.3%, PH is in the liquid nutrient medium Erlenmeyer flask of 7.5,32 DEG C, cultivate 72 hours under 220rpm oscillating condition, take out after substratum muddiness, then put into 2 DEG C of Refrigerator stores stand-by;
Blown-ball Azotobacter is transferred into glucose 20g/L, K
2hPO
41.0g/L, MgSO
40.2g/L, CaCO
30.5g/L, FeCl
30.25g/L, NaMoO
40.5g/L and with distilled water preparation pH be the liquid nutrient medium of 7.4, it is placed in the Erlenmeyer flask with granulated glass sphere, carries out shaking table cultivation, and rotating speed is 220rpm, cultivates 40h under 32 DEG C of conditions;
Rhodopseudomonas palustris is transferred into CH
3cOONa3.0g/L, NaHCO
35.0g/L, (NH
4)
2s0
41.0g/L, peptone 5.0g/L, yeast extract paste 1.0g/L, MgS0
40.5g/L, NaCl1.0g/L, KH
2p0
41.0g/L, K
2hPO
40.5g/L, CaCl
20.05g/L, CaCO
320g/L, pH are in the liquid nutrient medium of 7.3, leave standstill illumination (light intensity 2000lx), cultivate 6 days for 32 DEG C;
Step 2: many Pseudomonas bacterial classification is cultivated in synthesis domestication
(1) preparation of Microbe synthesis thalline suspension liquid
Join according to the weight proportion of Trichoderma 15 parts, Aspergillus candidus 15 parts, streptomycete 25 parts, subtilis 20 parts, Candida humicola 10 parts, blown-ball Azotobacter 15 parts and get each bacterial classification, and with the weight proportion of 1:100, described each bacterial classification is added in mixed-culture medium, put into fermentor tank, cultivate 6 days under the condition of the stirring velocity of 150rpm and 32 DEG C, until present filbert, obtain Microbe synthesis microbial inoculum, it is mixed with the 30wt% salt solution of 20 weight parts, the 20wt% Industry Waste molasses of 20 weight parts, obtains Microbe synthesis thalline suspension liquid; Wherein, described mixed-culture medium is by by wort 3 weight part, murphy juice 2.0 weight part, yeast extract paste 0.6 weight part, brown sugar 5 weight part, honey 3 weight part, MgSO
40.01 weight part, CaCO
30.02 weight part, FeCl
30.01 weight part, NaCl0.01 weight part, water 85.4 weight part mix, and use the ammoniacal liquor of 14wt% to regulate about pH to 7.2 to obtain;
(2) Rhodopseudomonas palustris spreads cultivation the preparation of liquid
Rhodopseudomonas palustris is added in basic medium, under natural lighting, 32 DEG C of conditions, spread cultivation 8 days, until in scarlet, obtain Rhodopseudomonas palustris and to spread cultivation liquid; Wherein, described basic medium is CH
3cOONa3.0g/L, NaHCO
35.0g/L, (NH
4)
2s0
41.0g/L, peptone 5.0g/L, yeast extract paste 1.0g/L, MgS0
40.5g/L, NaCl1.0g/L, KH
2p0
41.5g/L, K
2hPO
40.5g/L, CaCl
20.05g/L, CaCO
320g/L, pH are 7.0-7.5, and carbon-nitrogen ratio is the liquid nutrient medium of 1.5;
Step 3: preparation hybrid substrate
Will containing organic 12.5 weight parts, nitrogen 0.45 weight part, phosphorus (P
2o
5) 0.2 weight part, potassium (K
2o) 0.11 weight part and water content are that the cow dung of 80.1wr% processes in cow dung handler, are down to 55wt% to cow dung water content, tack-free, obtain cow dung of dewatering; Described dehydration cow dung 60 weight part, pig manure 20 weight part, stalk husk 17 weight part of pulverizing, diatomite 3 weight part are fully mixed, regulates water content to 35wt%, pH to 7.5, obtain described hybrid substrate;
Step 4: prepare microorganism problem soil ecology preparation for repairing standby
Spread cultivation Microbe synthesis thalline suspension liquid 15 weight part and Rhodopseudomonas palustris liquid 5 weight part, add in described hybrid substrate, stir, under the stirring velocity of 10rpm, carry out aerobic fermentation, wherein, earlier fermentation, the temperature namely in 5 days controls at 60 DEG C, and fermentation latter temperature controls at 35 DEG C, ferment 14 days, until fermentation material is grey, and with fermenting aroma, terminate fermentation; Fermentation material is dried or dried to water content lower than 10wt% under lower than 50 DEG C of conditions, and obtain described microorganism problem soil ecology preparation for repairing, wherein, total viable count is 10*10
8cfu/g, and harmless treat index reaches the standard-required of GB GB20287.
Application Example
Obtain in the restoration of the ecosystem of described microorganism problem soil ecology preparation for repairing for vegetables problematic soils in embodiment 1, for the plantation that organises of support facility vegetables, solve the continuous cropping obstacle that the problems such as soil disease increases the weight of, salinification cause, carry out the use of microorganism problem soil ecology preparation for repairing cucumber and tomato plastic greenhouse.While daily administration, during bulk application in early stage, base manure per ton adds microorganism problem soil ecology preparation for repairing described in 3kg, and after later stage field planting, every 1.5 months every is per muly evenly sprinkled into microorganism problem soil ecology preparation for repairing described in 1kg.The results are shown in following table:
Plastic tent cucumber and tomato soil Bacterial diversity quantity under different treatment
Visible, described microorganism problem soil ecology preparation for repairing is while improve soil microbial activities, also improve soil microflora, the increase of Population of Actinomycetes, be conducive to the increase of antibiotic and hormones biologically active substance in soil, certain restraining effect is risen to various soil-borne disease, alleviates continuous cropping obstacle.
Be described in detail specific embodiments of the invention above, but it is just as example, the present invention is not restricted to specific embodiment described above.To those skilled in the art, any equivalent modifications that the present invention is carried out and substituting also all among category of the present invention.Therefore, equalization conversion done without departing from the spirit and scope of the invention and amendment, all should contain within the scope of the invention.
Claims (10)
1. a microorganism problem soil ecology preparation for repairing, is characterized in that, is to adopt many Pseudomonas multi-cultur es synthesis domestication culture technique, and to comprise the hybrid substrate of dehydration cow dung for raw material, after carrying out solid fermentation by many Pseudomonas bacterial classification, drying and moulding obtains;
Wherein, the component of described many Pseudomonas bacterial classification and weight proportion thereof comprise:
2. microorganism problem soil ecology preparation for repairing according to claim 1, it is characterized in that, the water content of described hybrid substrate is 30-40wt%, pH is 7.0-9.0, and its component and weight proportion thereof comprise:
3. microorganism problem soil ecology preparation for repairing according to claim 1, is characterized in that, described auxiliary material be selected from diatomite, opoka, rock ash, spongolite any one or a few; Described stalk be selected from rice straw, maize straw, wheat stalk and other paddy wheat class stalk any one or a few.
4. a preparation method for the microorganism problem soil ecology preparation for repairing as described in claim 1-3 any one, it is characterized in that, described preparation method comprises:
Step 1: many Pseudomonas bacterial classification is cultivated in synthesis domestication
(1) preparation of Microbe synthesis thalline suspension liquid
Trichoderma, Aspergillus candidus, streptomycete, subtilis, Candida humicola, blown-ball Azotobacter bacterial classification are added in synthesis domestication substratum, under the stirring velocity, 30-60 DEG C condition of 150-200rpm, carry out synthesis domestication cultivate 5-7 days, obtain Microbe synthesis microbial inoculum, it is mixed with salt solution and protective material, obtains Microbe synthesis thalline suspension liquid;
(2) Rhodopseudomonas palustris spreads cultivation the preparation of liquid
Added by Rhodopseudomonas palustris in basic medium, under natural lighting, 28-35 DEG C condition, spread cultivation 8-12 days, until substratum is light red to scarlet, obtains Rhodopseudomonas palustris and to spread cultivation liquid;
Step 2: preparation hybrid substrate
Process cow dung makes its water content be down to 50-60wt%, obtains cow dung of dewatering; Described dehydration cow dung, pig manure, the stalk husk of pulverizing, auxiliary material are fully mixed, regulates water content to 30-40wt%, pH to 7.0-9.0, obtain described hybrid substrate;
Step 3: prepare microorganism problem soil ecology preparation for repairing standby
The liquid that Microbe synthesis thalline suspension liquid and Rhodopseudomonas palustris spread cultivation adds in described hybrid substrate, stir, and aerobic fermentation is carried out under the stirring velocity of 2-30rpm, wherein, earlier fermentation temperature controls be less than 65 DEG C, and fermentation latter temperature controls at 30-50 DEG C, fermentation 10-15 days, until fermentation material be light gray to Dark grey, and with fermenting aroma, terminate fermentation; Further fermentation material is dried to water content lower than 10wt% under lower than 50 DEG C of conditions, obtains described microorganism problem soil ecology preparation for repairing.
5. preparation method according to claim 4, it is characterized in that, described preparation method also comprises the preparation of each bacterial classification, wherein, by the Trichoderma (Trichodermasp. bought from Chinese agriculture Microbiological Culture Collection administrative center, ACCC31526), Aspergillus candidus (Aspergiluscandidus, ACCC30347), streptomycete (streptomycesStreptomycessp., ACCC41239), subtilis (Bacilussubtilis, ACCC01185), Candida humicola (Candidahumicola, ACCC20112), blown-ball Azotobacter (Azotobacterchroococcum, ACCC10003), Rhodopseudomonas palustris (Rhodopseudomonaspalustris, ACCC10649) carry out respectively cultivating each bacterial classification obtained for subsequent step.
6. preparation method according to claim 4, is characterized in that, described synthesis domestication substratum is by by wort 1-3 part, murphy juice 0.5-2.0 part, yeast extract paste 0.3-0.6 part, brown sugar 2-5 part, protective material 2-5 part, MgSO
40.01 part, CaCO
30.02 part, FeCl
30.01 part, NaCl0.01 part, the mixing of water 80-98 part, and regulate pH to 6.8-7.5 to obtain.
7. preparation method according to claim 4 or 6, is characterized in that, described protective material be selected from sucrose, glucose, honey, molasses any one or a few.
8. preparation method according to claim 4, it is characterized in that, described basic medium is CH
3cOONa3.0g/L, NaHCO
35.0g/L, (NH
4)
2s0
41.0g/L, peptone 5.0g/L, yeast extract paste 1.0g/L, MgS0
40.5g/L, NaCl1.0g/L, KH
2p0
41.5g/L, K
2hPO
40.5g/L, CaCl
20.05g/L, CaCO
320g/L, pH are 7.0-7.5, and carbon-nitrogen ratio is the liquid nutrient medium of 1.5.
9. preparation method according to claim 4, it is characterized in that, described preparation method specifically comprises the steps:
Step 1: many Pseudomonas bacterial classification is cultivated in synthesis domestication
(1) preparation of Microbe synthesis thalline suspension liquid
According to Trichoderma 10-25 part, Aspergillus candidus 10-25 part, streptomycete 20-30 part, subtilis 15-25 part, Candida humicola 10-25 part, the weight proportion of blown-ball Azotobacter 10-25 part is joined and is got each bacterial classification, and with the weight proportion of 1:80-120, described each bacterial classification is added in mixed-culture medium, put into fermentor tank, 5-7 days is cultivated under the condition of the stirring velocity of 150-200rpm and 30-60 DEG C, until present yellow to filbert, obtain Microbe synthesis microbial inoculum, by the 30wt% salt solution of itself and 20-40 weight part, the 20wt% Industry Waste molasses mixing of 20-40 weight part, obtain Microbe synthesis thalline suspension liquid, wherein, described mixed-culture medium is by by wort 1-3 weight part, murphy juice 0.5-2.0 weight part, yeast extract paste 0.3-0.6 weight part, brown sugar 2-5 weight part, honey 2-5 weight part, MgSO
40.01 weight part, CaCO
30.02 weight part, FeCl
30.01 weight part, NaCl0.01 weight part, water 80-98 weight part mix, and regulate pH to 6.8-7.5 to obtain,
(2) Rhodopseudomonas palustris spreads cultivation the preparation of liquid
Added by Rhodopseudomonas palustris in basic medium, under natural lighting, 28-35 DEG C condition, spread cultivation 8-12 days, until in light red to scarlet, obtain Rhodopseudomonas palustris and to spread cultivation liquid; Wherein, described basic medium is CH
3cOONa3.0g/L, NaHCO
35.0g/L, (NH
4)
2s0
41.0g/L, peptone 5.0g/L, yeast extract paste 1.0g/L, MgS0
40.5g/L, NaCl1.0g/L, KH
2p0
41.5g/L, K
2hPO
40.5g/L, CaCl
20.05g/L, CaCO
320g/L, pH are 7.0-7.5, and carbon-nitrogen ratio is the liquid nutrient medium of 1.5;
Step 2: preparation hybrid substrate
Will containing organic 11.0-14.5 weight part, nitrogen 0.30-0.45 weight part, phosphorus (P
2o
5) 0.15-0.25 weight part, potassium (K
2o) 0.10-0.15 weight part and water content are that the cow dung of 80.0-83.3wr% processes, and are down to 50-60wt% to cow dung water content, obtain cow dung of dewatering; Described dehydration cow dung 50-70 weight part, pig manure 10-20 weight part, the stalk husk 10-20 weight part of pulverizing, auxiliary material 0.5-3.5 weight part are fully mixed, regulates water content to 30-40wt%, pH to 7.0-9.0, obtain described hybrid substrate;
Step 3: prepare microorganism problem soil ecology preparation for repairing standby
Spread cultivation Microbe synthesis thalline suspension liquid 5-15 weight part and Rhodopseudomonas palustris liquid 1-10 weight part, add in described hybrid substrate, stir, under the stirring velocity of 2-30rpm, carry out aerobic fermentation, wherein, earlier fermentation, the temperature namely in 5 days controls be less than 65 DEG C, and fermentation latter temperature controls at 30-50 DEG C, fermentation 10-15 days, until fermentation material be light gray to Dark grey, and with fermenting aroma, terminate fermentation; Fermentation material is dried to water content lower than 10wt% under lower than 50 DEG C of conditions, obtains described microorganism problem soil ecology preparation for repairing.
10. the application of the microorganism problem soil ecology preparation for repairing as described in claim 1-3 any one.
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| CN113272254A (en) * | 2018-11-13 | 2021-08-17 | 南方海绵岩工业有限公司 | Liquid purification method |
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