CN103627658B - Microbial remediation agent for remediating soil ecosystem - Google Patents
Microbial remediation agent for remediating soil ecosystem Download PDFInfo
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- CN103627658B CN103627658B CN201310607191.3A CN201310607191A CN103627658B CN 103627658 B CN103627658 B CN 103627658B CN 201310607191 A CN201310607191 A CN 201310607191A CN 103627658 B CN103627658 B CN 103627658B
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- 239000003795 chemical substances by application Substances 0.000 title claims abstract description 64
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- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 12
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- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a microbial remediation agent for remediating a soil ecosystem. The microbial remediation agent is prepared by virtue of the following steps: uniformly mixing the following fermented bacteria at a viable count ratio: 50-60 parts of bacillus megatherium, 40-50 parts of bacillus licheniformis, 30-50 parts of clostridium butyricum, 20-50 parts of saccharomycetes, 1-2 parts of lactobacillus and 1 part of streptomycs micuoflaudio-videous so as to obtain a liquid microbial remediation agent, wherein bacillus megatherium, bacillus licheniformis, clostridium butyricum and saccharomycetes are used as main bacteria and lactobacillus and streptomycs micuoflaudio-videous are used as auxiliary bacteria; uniformly mixing the liquid microbial remediation agent with a solid adsorbate to obtain a solid microbial remediation agent. After entering the soil ecosystem, the microbial remediation agent disclosed by the invention can form a dominant bacterial population together with native-born beneficial microorganisms so as to promote nitrogen and oxygen circulation of the soil ecosystem, so that the soil ecosystem is remediated, a new stable balanced ecosystem is formed, the growing environment of crops is improved and the quality and yield of the crops are improved.
Description
Technical field
The invention belongs to soil ecosystem and repair field, particularly a kind of microbe soil restoration agent for soil ecosystem reparation and preparation method thereof, and the application of this microbe soil restoration agent in rehabilitating soil.
Background technology
Along with industrial and agricultural development, soil resource bears the increasing pressure, due to use and the heavy metal contamination of a large amount of chemical fertilizer, agricultural chemicals, radiocontamination makes the organism of difficult degradation in soil increasing, thus cause that soil microorganisms kind and quantity reduce, Soil structure and ecosystem destruction, crops quality decline, disease and pest increases, soil ecosystem is destroyed.
Summary of the invention
The object of the present invention is to provide a kind of microorganism renovation agent of the rehabilitating soil ecosystem;
Another object of the present invention is to provide a kind of method preparing the microorganism renovation agent of the rehabilitating soil ecosystem; Still a further object of the present invention is that above-mentioned microorganism renovation agent is applied to soil remediation, promotes plant growth.
The object of the invention is to be achieved through the following technical solutions, a kind of microorganism renovation agent of the rehabilitating soil ecosystem, it is characterized in that, with bacillus megaterium, Bacillus licheniformis, clostridium butylicum and yeast are as main body bacterium, with milk-acid bacteria, streptomyces microflavus is as auxiliary bacterium, the each bacterial classification fermented is mixed according to following viable count ratio and obtains liquid microbe renovation agent, bacillus megaterium 50-60, Bacillus licheniformis 40-50, clostridium butylicum 30-50, yeast 20-50, milk-acid bacteria 1-2, streptomyces microflavus 1, living bacteria count in mixed bacteria liquid >=300,000,000/milliliter.
Further, namely described liquid microbe renovation agent and solid absorption thing obtain solid microbe renovation agent after mixing according to the envelope-bulk to weight ratio of 1 liter: 5 ~ 10 kilograms.
Further, described solid absorption thing is the miscellany of turfy soil and vermiculite, obtained after pulverizing, high-temperature sterilization.
Further, the living bacteria count >=300,000,000/milliliter of described main body bacterium; Living bacteria count >=1,000 ten thousand/the milliliters of described auxiliary bacterium.
Prepare a method for the microorganism renovation agent of the above-mentioned rehabilitating soil ecosystem, comprise the following steps:
(1) respectively with solid medium recovery bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus, and mono-clonal bacterium colony is picked out;
(2) the mono-clonal bacterium colony of the bacillus megaterium picked out, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus is seeded in level liquid substratum respectively, obtains liquid spawn;
(3) respectively by the liquid spawn of bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus in secondary culture medium, amplification culture;
(4) respectively bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus are inoculated into fermentation cylinder for fermentation cultivation through the bacterium liquid of amplification culture, obtain producing and use bacterium liquid;
(5) use bacterium liquid according to viable count proportions production, mix and obtain liquid microbe renovation agent;
(6) turfy soil and vermiculite are pulverized, and through high-temperature sterilization, after being cooled to room temperature, are sprayed thereon by aforesaid liquid microorganism renovation agent, fully absorb after mixing and namely obtain solid microbe remediation microbial inoculum.
The application of microorganism renovation agent in rehabilitating soil of the above-mentioned rehabilitating soil ecosystem, comprises and executing or/and impose after crop-planting for base before crop-planting.
In mentioned microorganism renovation agent, various microbial fermentation solution all can conventionally be prepared, and the strain inoculation bought is carried out to solid medium recovery and cultivates, after thalli growth, and picking mono-clonal; Mono-clonal is inoculated in gobbet substratum and cultivates (one-level cultivation), obtain liquid spawn; Amplification culture (secondary cultivation) will be carried out in liquid-spawn inoculation to middle quantity of fluid substratum; Bacterial classification after amplification culture, being inoculated into fermentation cylinder for fermentation cultivation, finally obtains the liquid bacterial agent of wanted bacterial classification.For the microorganism related in this invention, can prepare with reference to following method:
Following various formulas are thalline liquid culture based formulas, and solid medium is on the basis of level liquid substratum, add 2% agar.In level liquid substratum, specifically add the agar of 2% exactly, be heated to 100 DEG C of dissolvings, cool at 40 DEG C and solidify, becoming solid state and be solid medium.All substratum are all 120oC autoclave sterilization process 30 minutes.
(1) bacillus megaterium (Bacillus megaterium, ATCC 14581, ATCC is numbered American type culture collection numbering):
(1) recovery activation, picking mono-clonal: by the strain inoculation bought in solid culture, 37 DEG C of quiescent culture 24 hours.The formula of solid culture is agar 2%, extractum carnis 0.5%, peptone 1%, sodium-chlor 0.2%, glucose 0.5%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.0.By said mixture, 120 DEG C of autoclave sterilization process 30 minutes, cool at 40 DEG C and solidify.
(2) one-level is cultivated, and obtains liquid spawn: by mono-clonal colony inoculation in level liquid substratum, and 37 DEG C of shaking tables are cultivated, and rotating speed is 200 revs/min, obtains liquid spawn after incubated overnight.Wherein level liquid culture medium prescription is extractum carnis 0.5%, peptone 1%, sodium-chlor 0.2%, glucose 0.5%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.0.Substratum was 120oC autoclave sterilization process 30 minutes.
(3) secondary cultivation, amplification culture: by liquid-spawn inoculation in secondary liquid substratum, 37 DEG C of shaking tables are cultivated, and rotating speed is 200 revs/min, spends the night, and carries out amplification culture.Wherein secondary liquid culture medium prescription is extractum carnis 1%, peptone 1%, sodium-chlor 0.2%, glucose 0.5%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.0.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(4) fermentation culture: carry out fermentation culture in strain inoculation amplification culture obtained to fermentor tank.Wherein fermentor cultivation based formulas is starch 0.5%, peptone 0.5%, sucrose 1%, sodium-chlor 0.2%, potassium primary phosphate 0.03%, dipotassium hydrogen phosphate 0.015%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.0.Substratum was 120oC autoclave sterilization process 30 minutes.
(2) Bacillus licheniformis (Bacillus licheniformis, ATCC 11946):
(1) recovery activation, picking mono-clonal: by the strain inoculation bought in solid culture, 37 DEG C of quiescent culture 24 hours.The formula of solid culture is agar 2%, extractum carnis 3%, peptone 1%, sodium-chlor 0.1%, glucose 0.5%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.5.By said mixture, 120oC autoclave sterilization process 30 minutes, cool at 40 DEG C and solidify.
(2) one-level is cultivated, and obtains liquid spawn: by mono-clonal colony inoculation in level liquid substratum, and 37 DEG C of shaking tables are cultivated, and rotating speed is 200 revs/min, obtains liquid spawn after incubated overnight.Wherein level liquid culture medium prescription is extractum carnis 3%, peptone 1%, sodium-chlor 0.1%, glucose 0.5%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.5.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(3) secondary cultivation, amplification culture: by liquid-spawn inoculation in secondary liquid substratum, 37 DEG C of shaking tables are cultivated, and rotating speed is 200 revs/min, spends the night, and carries out amplification culture.Wherein secondary liquid culture medium prescription is extractum carnis 1%, peptone 1%, sodium-chlor 0.1%, glucose 0.5%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.5.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(4) fermentation culture: carry out fermentation culture in strain inoculation amplification culture obtained to fermentor tank.Wherein fermentor cultivation based formulas is starch 0.5%, peptone 0.5%, soybean cake powder leaching juice 1%, sucrose 1%, sodium-chlor 0.2%, and potassium primary phosphate 0.03%, dipotassium hydrogen phosphate 0.02%, dissolve with distilled water, complement to 100% with water, adjust ph is 7.5.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(3) clostridium butylicum (Clostridium tyrobutyricum, ATCC 25755):
(1) recovery activation, picking mono-clonal: by the strain inoculation bought in solid culture, 37 DEG C of quiescent culture 24 hours.The formula of solid culture is agar 2%, extractum carnis 3%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, ammonium sulfate 0.01%, manganous sulfate 0.01%, magnesium sulfate 0.005%, and dissolve with distilled water, complement to 100% with water, adjust ph is 6.8.By said mixture, 120 DEG C of autoclave sterilization process 30 minutes, cool at 40 DEG C and solidify.
(2) one-level is cultivated, and obtain liquid spawn: by mono-clonal colony inoculation in level liquid substratum, 37 DEG C of cultivations, obtain liquid spawn.Wherein level liquid culture medium prescription is extractum carnis 3%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, ammonium sulfate 0.01%, manganous sulfate 0.01%, magnesium sulfate 0.005%, and dissolve with distilled water, complement to 100% with water, adjust ph is 6.8.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(3) secondary cultivation, amplification culture: by liquid-spawn inoculation in secondary liquid substratum, 37 DEG C are carried out amplification culture.Wherein secondary liquid culture medium prescription is extractum carnis 1%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, ammonium sulfate 0.01%, manganous sulfate 0.01%, magnesium sulfate 0.005%, and dissolve with distilled water, complement to 100% with water, adjust ph is 6.8.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(4) fermentation culture: carry out fermentation culture in strain inoculation amplification culture obtained to fermentor tank.Wherein fermentor cultivation based formulas is yeast powder 0.5%, peptone 0.5%, soybean cake powder leaching juice 1%, sucrose 1%, sodium-chlor 0.5%, ammonium sulfate 0.01%, manganous sulfate 0.01%, magnesium sulfate 0.005%, calcium carbonate 0.01%, potassium primary phosphate 0.02%, dipotassium hydrogen phosphate 0.01%, dissolve with distilled water, complement to 100% with water, adjust ph is 6.8.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(4) yeast (Saccharomyces cerevisiae, ATCC 16664):
(1) recovery activation, picking mono-clonal: by the strain inoculation of purchase in solid culture, 28 DEG C of quiescent culture.The formula of solid culture is agar 2%, yeast extract 0.5%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, and dissolve with distilled water, complement to 100% with water, adjust ph is 6.8.By said mixture, 120 DEG C of autoclave sterilization process 30 minutes, cool at 40 DEG C and solidify.
(2) one-level is cultivated, and obtain liquid spawn: by mono-clonal colony inoculation in level liquid substratum, 28 DEG C of cultivations, obtain liquid spawn.Wherein level liquid culture medium prescription is yeast extract 0.5%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, and dissolve with distilled water, complement to 100% with water, adjust ph is 6.8.Substratum was 120oC autoclave sterilization process 30 minutes.
(3) secondary cultivation, amplification culture: by liquid-spawn inoculation in secondary liquid substratum, 28 DEG C are carried out amplification culture.Wherein secondary liquid culture medium prescription is yeast extract 1%, peptone 1%, sodium-chlor 0.5%, glucose 0.5%, and dissolve with distilled water, complement to 100% with water, adjust ph is 6.8.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(4) fermentation culture: carry out fermentation culture in strain inoculation amplification culture obtained to fermentor tank.Wherein fermentor cultivation based formulas is yeast powder 0.5%, peptone 0.5%, soybean cake powder leaching juice 1%, sucrose 1%, sodium-chlor 0.5%, potassium primary phosphate 0.04%, dipotassium hydrogen phosphate 0.01%, calcium carbonate 0.1%, dissolve with distilled water, complement to 100% with water, adjust ph is 6.8.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(5) milk-acid bacteria (Lactobacillus plantarum, ATCC 8014):
(1) recovery activation, picking mono-clonal: by the strain inoculation bought in solid culture, 37 DEG C of quiescent culture 24 hours.The formula of solid culture is agar 2%, extractum carnis 0.5%, yeast extract 0.5%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, sodium acetate 0.05%, citric acid diamines 0.02%, ferrous sulfate 0.01%, potassium primary phosphate 0.03%, dipotassium hydrogen phosphate 0.02%, dissolve with distilled water, complement to 100% with water, adjust ph is 7.2.By said mixture, 120 DEG C of autoclave sterilization process 30 minutes, cool at 40 DEG C and solidify.
(2) one-level is cultivated, and obtain liquid spawn: by mono-clonal colony inoculation in level liquid substratum, 37 DEG C of cultivations, obtain liquid spawn.Wherein level liquid culture medium prescription is extractum carnis 0.5%, yeast extract 0.5%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, sodium acetate 0.05%, citric acid diamines 0.02%, ferrous sulfate 0.01%, potassium primary phosphate 0.03%, dipotassium hydrogen phosphate 0.02%, dissolves with distilled water, complement to 100% with water, adjust ph is 7.2.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(3) secondary cultivation, amplification culture: by liquid-spawn inoculation in secondary liquid substratum, 37 DEG C are carried out amplification culture.Wherein secondary liquid culture medium prescription is extractum carnis 1%, yeast extract 0.5%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, sodium acetate 0.05%, citric acid diamines 0.02%, ferrous sulfate 0.01%, potassium primary phosphate 0.03%, dipotassium hydrogen phosphate 0.02%, dissolves with distilled water, complement to 100% with water, adjust ph is 7.2.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(4) fermentation culture: carry out fermentation culture in strain inoculation amplification culture obtained to fermentor tank.Wherein fermentor cultivation based formulas is Zulkovsky starch 0.5%, yeast extract 0.5%, peptone 0.5%, sucrose 1%, sodium-chlor 0.5%, sodium acetate 0.05%, citric acid diamines 0.02%, ferrous sulfate 0.01%, potassium primary phosphate 0.03%, dipotassium hydrogen phosphate 0.02%, dissolves with distilled water, complement to 100% with water, adjust ph is 6.8.Substratum was 120oC autoclave sterilization process 30 minutes.
(6) streptomyces microflavus (Streptomyces microflavus, ATCC 23877):
(1) recovery activation, picking mono-clonal: by the strain inoculation bought in solid culture, 37 DEG C of quiescent culture 24 hours.The formula of solid culture is extractum carnis 0.5%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, potassium primary phosphate 0.03%, dipotassium hydrogen phosphate 0.02%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.0.By said mixture, 120 DEG C of autoclave sterilization process 30 minutes, cool at 40 DEG C and solidify.
(2) one-level is cultivated, and obtain liquid spawn: by mono-clonal colony inoculation in level liquid substratum, 37 DEG C of cultivations, obtain liquid spawn.Wherein level liquid culture medium prescription is extractum carnis 0.5%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, potassium primary phosphate 0.03%, dipotassium hydrogen phosphate 0.02%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.0.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(3) secondary cultivation, amplification culture: by liquid-spawn inoculation in secondary liquid substratum, 37 DEG C are carried out amplification culture.Wherein secondary liquid culture medium prescription is extractum carnis 1%, peptone 1%, sodium-chlor 0.5%, glucose 0.3%, potassium primary phosphate 0.03%, dipotassium hydrogen phosphate 0.02%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.0.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
(4) fermentation culture: carry out fermentation culture in strain inoculation amplification culture obtained to fermentor tank.Wherein fermentor cultivation based formulas is Zulkovsky starch 0.5%, peptone 0.5%, sucrose 1%, sodium-chlor 0.5%, sodium acetate 0.05%, potassium primary phosphate 0.03%, dipotassium hydrogen phosphate 0.02%, and dissolve with distilled water, complement to 100% with water, adjust ph is 7.2.Substratum was 120 DEG C of autoclave sterilization process 30 minutes.
Microorganism renovation agent of the present invention is after entering soil ecosystem, jointly dominant microflora can be formed with the beneficial microorganism of original inhabitants, promote the nitrogen of soil ecosystem, oxygen cycle, thus reach the rehabilitating soil ecosystem, make it to form new stable balanced ecosystem, improve the growing environment of farm crop, improve the Quality and yield of farm crop.
Embodiment
Below in conjunction with specific embodiment, the present invention is further illustrated.
The preparation method of the microorganism renovation agent of a kind of rehabilitating soil ecosystem that the present invention mentions, comprises the following steps:
(1) respectively with solid medium recovery bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus, mono-clonal bacterium colony is selected;
(2) respectively bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus are seeded in level liquid substratum, obtain liquid spawn;
(3) respectively by the liquid spawn of bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus in secondary culture medium, amplification culture;
(4) respectively the bacterium liquid of the amplification culture of bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus is inoculated into fermentation cylinder for fermentation to cultivate, obtains producing and use bacterium liquid;
(5) bacterium liquid is proportionally mixed to get liquid microbe renovation agent;
(6) liquid microbe renovation agent and the porous mass such as turfy soil and vermiculite are obtained solid microbe remediation microbial inoculum.
Embodiment 1:
1, liquid microbe renovation agent is prepared: mixed according to following viable count ratio by each bacterial classification fermented, bacillus megaterium: Bacillus licheniformis: clostridium butylicum: yeast: milk-acid bacteria: streptomyces microflavus=50:50:50:50:1:1; Mix, namely obtain liquid microbe renovation agent;
2, turfy soil and vermiculite are pulverized, after high-temperature sterilization, as solid absorption thing;
3, by 1 part of volume (unit: rise) liquid microbe renovation agent, be sprayed on the solid absorption thing of 5 parts of quality (unit: kilogram), mix, namely obtain solid microbe renovation agent.
Embodiment 2:
1, liquid microbe renovation agent is prepared: mixed according to following viable count ratio by each bacterial classification fermented, bacillus megaterium: Bacillus licheniformis: clostridium butylicum: yeast: milk-acid bacteria: streptomyces microflavus=50:50:30:30:1:1; Mix, namely obtain liquid microbe renovation agent;
2, turfy soil and vermiculite are pulverized, after high-temperature sterilization, as solid absorption thing;
3, by 1 part of volume (unit: rise) liquid microbe renovation agent, be sprayed on the solid absorption thing of 7.5 parts of quality (unit: kilogram), mix, namely obtain solid microbe renovation agent.
Embodiment 3:
1, liquid microbe renovation agent is prepared: mixed according to following viable count ratio by each bacterial classification fermented, bacillus megaterium: Bacillus licheniformis: clostridium butylicum: yeast: milk-acid bacteria: streptomyces microflavus=60:40:40:20:2:1; Mix, namely obtain liquid microbe renovation agent;
2, turfy soil and vermiculite are pulverized, after high-temperature sterilization, as solid absorption thing;
3, by 1 part of volume (unit: rise) liquid microbe renovation agent, be sprayed on the solid absorption thing of 10 parts of quality (unit: kilogram), mix, namely obtain solid microbe renovation agent.
Test example 1: the effect test of solid microbe renovation agent of the present invention
1, test sample: the microorganism renovation agent prepared by embodiment 1-3
2, test method: this test utilizes potted plant growth rape to complete in greenhouse.Concrete steps are as follows: rape club root old complaint grinds by (1), and mix with the ratio of Nutrition Soil according to weight ratio 1:50, obtain bacterium earth mixtures; (2) bacterium earth mixtures is divided into 4 groups, first, second and third organizes the microorganism renovation agent added respectively prepared by embodiment 1,2,3, and additional proportion is the bacterium earth mixtures that 1 part of microorganism renovation agent joins 50 parts, mixes; 4th group does not add any microorganism renovation agent and other material, in contrast; (3) in four groups, sow 50 Semen Brassicae campestriss respectively, wait to germinate two days later, often organize thinning, leave 36; (4) four groups of rapes are cultivated in greenhouse, and the photoperiod is 16L:8D (illumination 16 hours, unglazed photograph 8 hours), relative humidity 70%, and greenhouse room temperature is 30 DEG C; (5) after germinateing 6 weeks, band root collects rape, the incidence of root club root is measured after removing root soil, and measure plant height, the over-ground part fresh weight of plant, calculating mean value and standard error, carry out statistically otherness by one-way analysis of variance (one-way ANOVA) method significantly to analyze, during setting P<0.05, have significant difference.
3, test-results: the incidence of club root carries out counting statistics analysis according to 5 grades of methods: 0 grade, normal, the disease-free symptom of root system; 1 grade, the indivedual tiny root nodule of fibrous root, side root, there is not disease in main root; 3 grades, fibrous root, the more root nodule of side root or root nodule diameter or length are more than 0.5 centimetre, or main root occurs Disease symptoms but enlargement is not obvious; 5 grades, main root enlargement is obvious, and tumescence diameter is stem foot 2-3 times; 7 grades, main root enlargement is obvious, and tumescence diameter is stem foot more than 3 times.Disease index is according to following formulae discovery: disease index=［ Σ (diseased plant number × appropriate level at different levels) ÷ (investigating total strain number × morbidity highest level typical value) ］ × 100.Test-results shows, compared with morbidity control group (the 4th group), application of first, second and third group of microorganism renovation agent of the present invention, sickness rate and the disease index of rape club root all significantly decline, and preventing effectiveness reaches more than 68%; The rape plant height and the fresh weight comparatively morbidity group that meanwhile application of microorganism renovation agent increase significantly, and concrete data see the following form (numerical value of plant height and fresh weight is mean+/-standard error); And there is no significant difference between first, second and third group.
| Process | Sickness rate | Disease index | Prevention effect | Plant height (cm) | Fresh weight (g) |
| First group | 69.4% | 27.4% | 72.6% | 10.9 ±0.8 | 2.58 ± 0.23 |
| Second group | 63.9% | 31.3% | 68.7% | 11.8 ± 0.9 | 2.77 ± 0.24 |
| 3rd group | 72.2% | 29.4% | 70.6% | 11.5 ± 0.7 | 2.50 ± 0.20 |
| 4th group (morbidity control group) | 100.0% | 92.1% | - | 5.5 ± 0.3 | 1.42 ± 0.15 |
Test example 2: the effect test of liquid microbe renovation agent of the present invention
1, test sample: the microorganism renovation agent (liquid) prepared by embodiment 1
2, test method: this test utilizes potted plant growth rape to complete in greenhouse.Testing sequence is basic identical with test example 1.But only have two groups, often organize 24 Brassica campestris L seedlings and be used for test.The plantation group that first group (reparation group) is applicating liquid microorganism renovation agent, second group (morbidity control group) is morbidity control group.After germination, according to the method for test example 1, measure plant height, fresh weight, record incidence.Wherein between two groups, the otherness of plant height and fresh weight adopts student t to check (Student's t-test) method, has significant difference during setting P<0.05.
3, test-results: reparation group (first group) is compared with morbidity control group (second group), and application of first group of liquid microbe renovation agent of the present invention, sickness rate and the disease index of rape club root all significantly decline, and preventing effectiveness reaches more than 68%; The rape plant height and the fresh weight comparatively morbidity group that meanwhile application of microorganism renovation agent increase significantly (P<0.05).
| Process | Sickness rate | Disease index | Prevention effect | Plant height (cm) | Fresh weight (g) |
| First group (reparation group) | 70.8% | 31.5% | 68.5.6% | 10.7 ± 0.5 | 2.26 ± 0.18 |
| Second group (morbidity control group) | 100.0% | 91.7% | - | 6.5 ± 0.3 | 1.33 ± 0.12 |
The above is only preferred embodiment of the present invention, not does any pro forma restriction to the present invention; Any those of ordinary skill in the art, do not departing under technical solution of the present invention ambit, the Method and Technology content of above-mentioned announcement all can be utilized to make many possible variations and modification to technical solution of the present invention, or be revised as the Equivalent embodiments of equivalent variations.Therefore, every content not departing from technical solution of the present invention, according to technical spirit of the present invention to any simple modification made for any of the above embodiments, equivalent replacement, equivalence change and modification, all still belongs in the scope of technical solution of the present invention protection.
Claims (7)
1. the microorganism renovation agent of a rehabilitating soil ecosystem, it is characterized in that, using bacillus megaterium, Bacillus licheniformis, clostridium butylicum and yeast as main body bacterium, using milk-acid bacteria, streptomyces microflavus as auxiliary bacterium, the each bacterial classification fermented is mixed according to following viable count ratio and obtains liquid microbe renovation agent, bacillus megaterium 50-60, Bacillus licheniformis 40-50, clostridium butylicum 30-50, yeast 20-50, milk-acid bacteria 1-2, streptomyces microflavus 1, the living bacteria count in mixed bacteria liquid >=300,000,000/milliliter; Described yeast is yeast saccharomyces cerevisiae, and described milk-acid bacteria is plant lactobacillus.
2. the microorganism renovation agent of a kind of rehabilitating soil ecosystem according to claim 1, it is characterized in that, namely described liquid microbe renovation agent and solid absorption thing obtain solid microbe renovation agent after mixing according to the envelope-bulk to weight ratio of 1 liter: 5 ~ 10 kilograms.
3. the microorganism renovation agent of a kind of rehabilitating soil ecosystem according to claim 2, is characterized in that, described solid absorption thing is the mixture of turfy soil and vermiculite, obtained after pulverizing, high-temperature sterilization.
4. the microorganism renovation agent of a kind of rehabilitating soil ecosystem according to claim 1, is characterized in that, the living bacteria count >=300,000,000/milliliter of described main body bacterium; Living bacteria count >=1,000 ten thousand/the milliliters of described auxiliary bacterium.
5. prepare a method for the microorganism renovation agent of the rehabilitating soil ecosystem described in claim 1 or 4, it is characterized in that, comprise the following steps:
(1) respectively with solid medium recovery bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus, and mono-clonal bacterium colony is picked out;
(2) the mono-clonal bacterium colony of the bacillus megaterium picked out, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus is seeded in level liquid substratum respectively, obtains liquid spawn;
(3) respectively by the liquid spawn of bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus in secondary culture medium, amplification culture;
(4) respectively bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus are inoculated into fermentation cylinder for fermentation cultivation through the bacterium liquid of amplification culture, obtain producing and use bacterium liquid;
(5) use bacterium liquid according to viable count proportions in claim 1 production, mix and obtain liquid microbe renovation agent.
6. prepare a method for the microorganism renovation agent of the rehabilitating soil ecosystem described in Claims 2 or 3, it is characterized in that, comprise the following steps:
(1) respectively with solid medium recovery bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus, and mono-clonal bacterium colony is picked out;
(2) the mono-clonal bacterium colony of the bacillus megaterium picked out, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus is seeded in level liquid substratum respectively, obtains liquid spawn;
(3) respectively by the liquid spawn of bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus in secondary culture medium, amplification culture;
(4) respectively bacillus megaterium, Bacillus licheniformis, clostridium butylicum, yeast, milk-acid bacteria and streptomyces microflavus are inoculated into fermentation cylinder for fermentation cultivation through the bacterium liquid of amplification culture, obtain producing and use bacterium liquid;
(5) use bacterium liquid according to viable count proportions in claim 1 production, mix and obtain liquid microbe renovation agent;
(6) turfy soil and vermiculite are pulverized, and through high-temperature sterilization, after being cooled to room temperature, are sprayed thereon by aforesaid liquid microorganism renovation agent, fully absorb after mixing and namely obtain solid microbe remediation microbial inoculum.
7. the application of microorganism renovation agent in rehabilitating soil control rape club root of the arbitrary described rehabilitating soil ecosystem of claim 1-4.
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| TWI604794B (en) * | 2016-12-09 | 2017-11-11 | Yeast composition | |
| CN110468059A (en) * | 2018-05-12 | 2019-11-19 | 哈尔滨大自然环境工程有限公司 | A kind of liquid aerobic microbiological soil-repairing agent and its production application method |
| CN108728108B (en) * | 2018-05-30 | 2021-02-02 | 山东绿之行环境工程有限公司 | Medicament for repairing contaminated soil and preparation method thereof |
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