CN108728383B - A kind of preparation method of probiotics and its application in degrading pesticide residues - Google Patents
A kind of preparation method of probiotics and its application in degrading pesticide residues Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 33
- 239000000447 pesticide residue Substances 0.000 title claims abstract description 9
- 230000000593 degrading effect Effects 0.000 title claims abstract description 4
- 239000006041 probiotic Substances 0.000 title abstract description 5
- 235000018291 probiotics Nutrition 0.000 title abstract description 5
- 238000004108 freeze drying Methods 0.000 claims abstract description 34
- 239000000047 product Substances 0.000 claims abstract description 33
- 239000000758 substrate Substances 0.000 claims abstract description 25
- 239000007787 solid Substances 0.000 claims abstract description 23
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 22
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 22
- 239000007853 buffer solution Substances 0.000 claims abstract description 19
- 239000002244 precipitate Substances 0.000 claims abstract description 17
- 108010059892 Cellulase Proteins 0.000 claims abstract description 16
- 229940106157 cellulase Drugs 0.000 claims abstract description 16
- 239000003223 protective agent Substances 0.000 claims abstract description 16
- 241000609240 Ambelania acida Species 0.000 claims abstract description 14
- 239000010905 bagasse Substances 0.000 claims abstract description 14
- 238000002791 soaking Methods 0.000 claims abstract description 12
- 238000001914 filtration Methods 0.000 claims abstract description 10
- 239000002504 physiological saline solution Substances 0.000 claims abstract description 10
- 239000006228 supernatant Substances 0.000 claims abstract description 10
- 239000000725 suspension Substances 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 5
- 238000002156 mixing Methods 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 16
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 12
- 238000001035 drying Methods 0.000 claims description 12
- 239000000575 pesticide Substances 0.000 claims description 10
- 238000010298 pulverizing process Methods 0.000 claims description 10
- 238000007710 freezing Methods 0.000 claims description 9
- 230000008014 freezing Effects 0.000 claims description 9
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 4
- 235000020183 skimmed milk Nutrition 0.000 claims description 4
- 239000005874 Bifenthrin Substances 0.000 claims description 3
- OMFRMAHOUUJSGP-IRHGGOMRSA-N bifenthrin Chemical compound C1=CC=C(C=2C=CC=CC=2)C(C)=C1COC(=O)[C@@H]1[C@H](\C=C(/Cl)C(F)(F)F)C1(C)C OMFRMAHOUUJSGP-IRHGGOMRSA-N 0.000 claims description 3
- BIWJNBZANLAXMG-YQELWRJZSA-N chloordaan Chemical compound ClC1=C(Cl)[C@@]2(Cl)C3CC(Cl)C(Cl)C3[C@]1(Cl)C2(Cl)Cl BIWJNBZANLAXMG-YQELWRJZSA-N 0.000 claims description 3
- 230000000694 effects Effects 0.000 claims description 3
- XQUXKZZNEFRCAW-UHFFFAOYSA-N fenpropathrin Chemical compound CC1(C)C(C)(C)C1C(=O)OC(C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 XQUXKZZNEFRCAW-UHFFFAOYSA-N 0.000 claims description 3
- 239000003993 organochlorine pesticide Substances 0.000 claims description 3
- 239000002728 pyrethroid Substances 0.000 claims description 3
- YVGGHNCTFXOJCH-UHFFFAOYSA-N DDT Chemical compound C1=CC(Cl)=CC=C1C(C(Cl)(Cl)Cl)C1=CC=C(Cl)C=C1 YVGGHNCTFXOJCH-UHFFFAOYSA-N 0.000 claims description 2
- 108090000790 Enzymes Proteins 0.000 claims 1
- 102000004190 Enzymes Human genes 0.000 claims 1
- 229940088598 enzyme Drugs 0.000 claims 1
- 238000007605 air drying Methods 0.000 abstract description 7
- 230000000529 probiotic effect Effects 0.000 abstract description 3
- 239000003337 fertilizer Substances 0.000 description 8
- 239000002689 soil Substances 0.000 description 7
- 235000012054 meals Nutrition 0.000 description 6
- 230000000813 microbial effect Effects 0.000 description 6
- 239000003895 organic fertilizer Substances 0.000 description 4
- 235000017060 Arachis glabrata Nutrition 0.000 description 3
- 244000105624 Arachis hypogaea Species 0.000 description 3
- 235000010777 Arachis hypogaea Nutrition 0.000 description 3
- 235000018262 Arachis monticola Nutrition 0.000 description 3
- 235000019779 Rapeseed Meal Nutrition 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 235000020232 peanut Nutrition 0.000 description 3
- 239000004456 rapeseed meal Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 description 2
- 235000000536 Brassica rapa subsp pekinensis Nutrition 0.000 description 2
- 241000499436 Brassica rapa subsp. pekinensis Species 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000005067 remediation Methods 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 240000002791 Brassica napus Species 0.000 description 1
- 235000004977 Brassica sinapistrum Nutrition 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 235000019764 Soybean Meal Nutrition 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000004455 soybean meal Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/04—Pesticides, e.g. insecticides, herbicides, fungicides or nematocides
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/28—Organic substances containing oxygen, sulfur, selenium or tellurium, i.e. chalcogen
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- Microbiology (AREA)
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- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Emergency Management (AREA)
- Fertilizers (AREA)
- Environmental & Geological Engineering (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- General Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Soil Sciences (AREA)
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- Chemical Kinetics & Catalysis (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明涉及一种微生态制剂的制备方法及其在降解农药残留中的应用，所述微生态制剂的制备方法，包括如下步骤：(1)将甘蔗渣自然风干至恒重，粉碎后，浸泡于质量分数为5‑8％的纤维素酶缓冲溶液中，pH为4.5‑6.0，浸泡时间为20‑24小时后，过滤得沉淀，沉淀干燥即得底物；(2)将枯草芽孢杆菌种子液与步骤(1)制备的底物混合均匀，于20‑40℃下发酵5‑7天后得发酵物，将发酵物离心后，弃去上清液，收集固体、用生理盐水洗涤后，与冻干保护剂混匀得混悬液，于‑70℃下预冻5h后，转移至冷冻干燥机中冷冻干燥24h后，即得所述微生态制剂。The invention relates to a preparation method of a microecological preparation and its application in degrading pesticide residues. The preparation method of the microecological preparation comprises the following steps: (1) naturally air-drying bagasse to constant weight, crushing, soaking In a cellulase buffer solution with a mass fraction of 5-8%, the pH is 4.5-6.0, and the soaking time is 20-24 hours, filtering to obtain a precipitate, and the precipitate is dried to obtain a substrate; (2) Bacillus subtilis seeds The liquid is evenly mixed with the substrate prepared in step (1), fermented at 20-40 ° C for 5-7 days to obtain a fermented product, after the fermented product is centrifuged, the supernatant is discarded, the solid is collected, washed with physiological saline, and then mixed with the fermented product. The freeze-drying protective agent is mixed to obtain a suspension, which is pre-frozen at -70°C for 5 hours, and then transferred to a freeze dryer for freeze-drying for 24 hours to obtain the probiotic preparation.
Description
Technical Field
The invention belongs to the field of microecologics, and particularly relates to a preparation method of a microecologic preparation and application of the microecologic preparation in pesticide residue degradation.
Background
China is a big agricultural country, and a large amount of pesticides and chemical fertilizers are used every year, so that although the use of the pesticides and the chemical fertilizers can prevent plant diseases and insect pests and promote the growth of crops, a large amount of pesticide and chemical fertilizer residues are caused, and the living environment and the body health of people are influenced. In recent years, with the continuous improvement of the requirements of life quality of people, safe and pollution-free green food is more and more popular among people, and ecological organic agriculture is more and more emphasized. The invention provides a microecological preparation which can effectively promote the growth of crops and can effectively degrade pesticide residues in soil.
Disclosure of Invention
The invention provides a microecological preparation, which is characterized in that the preparation method of the microecological preparation comprises the following steps:
(1) naturally drying bagasse to constant weight, crushing, soaking in 5-8 wt% cellulase buffer solution at pH of 4.5-6.0 for 20-24 hr, filtering to obtain precipitate, and drying the precipitate to obtain substrate;
(2) uniformly mixing the bacillus subtilis seed solution with the substrate prepared in the step (1), fermenting for 5-7 days at 20-40 ℃ to obtain a fermented product, centrifuging the fermented product, removing a supernatant, collecting a solid, washing with physiological saline, uniformly mixing with a freeze-drying protective agent to obtain a suspension, pre-freezing for 5 hours at-70 ℃, transferring to a freeze-drying machine, and freeze-drying for 24 hours to obtain the microecological preparation.
The pulverization in the step (1) is preferably pulverized to 40-60 meshes; the buffer solution is preferably citric acid-sodium citrate buffer solution; the activity of the cellulase is preferably 10-400KU/g, and the mass ratio of the bagasse to the cellulase buffer solution is 1: 8-10.
In the step (2), the mass ratio of the bacillus subtilis seed solution to the substrate is 10: 1-2; 1L of freeze-drying protective agent is used for each kilogram of bacillus subtilis seed liquid; each liter of freeze-drying protective agent contains 90g of glucose, 125g of skim milk powder, 16mL of glycerol and the balance of water.
Another embodiment of the present invention provides a method for preparing the above-mentioned microecological preparation, which is characterized by comprising the steps of:
(1) naturally drying bagasse to constant weight, crushing, soaking in 5-8 wt% cellulase buffer solution at pH of 4.5-6.0 for 20-24 hr, filtering to obtain precipitate, and drying the precipitate to obtain substrate;
(2) uniformly mixing the bacillus subtilis seed solution with the substrate prepared in the step (1), fermenting for 5-7 days at 20-40 ℃ to obtain a fermented product, centrifuging the fermented product, removing a supernatant, collecting a solid, washing with physiological saline, uniformly mixing with a freeze-drying protective agent to obtain a suspension, pre-freezing for 5 hours at-70 ℃, transferring to a freeze-drying machine, and freeze-drying for 24 hours to obtain the microecological preparation.
The pulverization in the step (1) is preferably pulverized to 40-60 meshes; the buffer solution is preferably citric acid-sodium citrate buffer solution; the activity of the cellulase is preferably 10-400KU/g, and the mass ratio of the bagasse to the cellulase buffer solution is 1: 8-10.
In the step (2), the mass ratio of the bacillus subtilis seed solution to the substrate is 10: 1-2; 1L of freeze-drying protective agent is used for each kilogram of bacillus subtilis seed liquid; each liter of freeze-drying protective agent contains 90g of glucose, 125g of skim milk powder, 16mL of glycerol and the balance of water.
Another embodiment of the present invention provides the use of the above-described probiotics in soil remediation.
Another embodiment of the present invention provides the use of the above-described probiotic for degrading pesticide residues. The pesticide is preferably organochlorine pesticide or pyrethroid pesticide, and is further preferably chlordane, DDT, fenpropathrin and bifenthrin.
Another embodiment of the present invention provides the use of the above-described probiotic for the preparation of a soil remediation agent.
The invention also provides a microbial organic fertilizer which is characterized by comprising the microbial organic fertilizer and oil meal in a mass ratio of 1: 5-8. The oil meal is selected from one or more of soybean meal, peanut meal or rapeseed meal.
Another embodiment of the invention provides an application of the microbial ecological agent in preparation of a microbial organic fertilizer.
The bacillus subtilis seed solution is prepared according to a conventional seed solution preparation method in the field, and comprises the following specific steps: inoculating slant strain of Bacillus subtilis into seed culture medium, and shake culturing at 20-25 deg.C for 24 hr to obtain seed solution; wherein each liter of seed culture medium contains 20g of glucose, 2g of peptone, 2g of yeast extract and the balance of water; the inoculation amount of the bacillus subtilis slant strains is 5-10%; the rotating speed of the shaking table is 100-120 rpm; the bacterial content in the seed liquid per milliliter is more than 10 hundred million.
The bacillus subtilis can be purchased through China general microbiological culture Collection center (CGMCC), preferably any one or a mixture of strains with CGMCC numbers of 1.9083, 1.8955, 1.8886 and 1.8801, and further preferably 1.8955 and 1.8886.
Compared with the prior art, the invention has the advantages that: (1) the method comprises the steps of treating bagasse by using cellulase to obtain a substrate, mixing the substrate with a bacillus subtilis seed solution, and fermenting to obtain a microecological preparation, wherein the microecological preparation can effectively degrade pesticide residues (especially pyrethroid pesticides and organochlorine pesticides) in soil; (2) the microbial ecological agent prepared by the invention can be mixed with oil meal to be used as a microbial organic fertilizer, can effectively increase the yield of Chinese cabbage, and is equivalent to the yield increase of 33-36% per mu of common compound fertilizer (N: P: K: 15: 15).
Detailed Description
In order to facilitate a further understanding of the invention, the following examples are provided to illustrate it in more detail. However, these examples are only for better understanding of the present invention and are not intended to limit the scope or the principle of the present invention, and the embodiments of the present invention are not limited to the following.
The lyoprotectant used in the following examples is the same, and each liter of lyoprotectant contains 90g of glucose, 125g of skim milk powder, 16mL of glycerol, and the balance of water.
Example 1
(1) Naturally air drying bagasse to constant weight (10.0kg), pulverizing to 40-60 mesh, soaking in 8% cellulase (10KU/g) citric acid-sodium citrate buffer solution (100.0kg) at pH of 4.5-5.0 for 24 hr, filtering to obtain precipitate, and drying to obtain substrate;
(2) uniformly mixing a bacillus subtilis (CGMCC 1.8955) seed solution (10.0kg) and a substrate (1.0kg) prepared in the step (1), standing and fermenting at 30-35 ℃ for 5 days to obtain a fermented product, centrifuging the fermented product, removing a supernatant, collecting a solid, washing the solid with physiological saline, uniformly mixing the solid with a freeze-drying protective agent (10L) to obtain a suspension, pre-freezing at-70 ℃ for 5h, transferring to a freeze-drying machine, and freeze-drying for 24h to obtain the microecological preparation (hereinafter referred to as a product A).
Example 2
(1) Naturally air drying bagasse to constant weight (10.0kg), pulverizing to 40-60 mesh, soaking in 5% cellulase (400KU/g) citric acid-sodium citrate buffer solution (80.0kg) at pH of 5.0-6.0 for 20 hr, filtering to obtain precipitate, and drying the precipitate to obtain substrate;
(2) uniformly mixing a bacillus subtilis (CGMCC 1.8886) seed solution (10.0kg) and a substrate (2.0kg) prepared in the step (1), standing and fermenting at 20-25 ℃ for 7 days to obtain a fermented product, centrifuging the fermented product, removing a supernatant, collecting a solid, washing the solid with physiological saline, uniformly mixing the solid with a freeze-drying protective agent (10L) to obtain a suspension, pre-freezing at-70 ℃ for 5h, transferring to a freeze-drying machine, and freeze-drying for 24h to obtain the microecological preparation (hereinafter referred to as a product B).
Example 3
(1) Naturally air drying bagasse to constant weight (10.0kg), pulverizing to 40-60 mesh, soaking in citric acid-sodium citrate buffer solution (100.0kg), pH 4.5-5.0, soaking for 24 hr, filtering to obtain precipitate, and drying the precipitate to obtain substrate;
(2) uniformly mixing a bacillus subtilis (CGMCC 1.8955) seed solution (10.0kg) and a substrate (1.0kg) prepared in the step (1), standing and fermenting at 30-35 ℃ for 5 days to obtain a fermented product, centrifuging the fermented product, removing a supernatant, collecting a solid, washing the solid with physiological saline, uniformly mixing the solid with a freeze-drying protective agent (10L) to obtain a suspension, pre-freezing at-70 ℃ for 5h, transferring to a freeze-drying machine, and freeze-drying for 24h to obtain a product C.
Example 4
(1) Naturally air drying bagasse to constant weight (10.0kg), pulverizing to 40-60 mesh, soaking in 8% cellulase (10KU/g) citric acid-sodium citrate buffer solution (100.0kg) at pH of 4.5-5.0 for 24 hr, filtering to obtain precipitate, and drying to obtain substrate;
(2) uniformly mixing a bacillus subtilis (CGMCC 1.8801) seed solution (10.0kg) and a substrate (1.0kg) prepared in the step (1), standing and fermenting at 30-35 ℃ for 5 days to obtain a fermented product, centrifuging the fermented product, removing a supernatant, collecting a solid, washing the solid with physiological saline, uniformly mixing the solid with a freeze-drying protective agent (10L) to obtain a suspension, pre-freezing at-70 ℃ for 5h, transferring to a freeze-drying machine, and freeze-drying for 24h to obtain the microecological preparation (hereinafter referred to as a product D).
Example 5
(1) Naturally air drying bagasse to constant weight (10.0kg), pulverizing to 40-60 mesh, soaking in 8% cellulase (10KU/g) citric acid-sodium citrate buffer solution (100.0kg) at pH of 4.5-5.0 for 24 hr, filtering to obtain precipitate, and drying to obtain substrate;
(2) uniformly mixing a bacillus subtilis (CGMCC 1.9083) seed solution (10.0kg) and a substrate (1.0kg) prepared in the step (1), standing and fermenting at 30-35 ℃ for 5 days to obtain a fermented product, centrifuging the fermented product, removing a supernatant, collecting a solid, washing the solid with physiological saline, uniformly mixing the solid with a freeze-drying protective agent (10L) to obtain a suspension, pre-freezing at-70 ℃ for 5h, transferring to a freeze-drying machine, and freeze-drying for 24h to obtain the microecological preparation (hereinafter referred to as a product E).
Example 6
(1) Naturally air drying bagasse to constant weight (10.0kg), pulverizing to 40-60 mesh, soaking in 5% cellulase (400KU/g) citric acid-sodium citrate buffer solution (80.0kg) at pH of 5.0-6.0 for 20 hr, filtering to obtain precipitate, and drying the precipitate to obtain substrate;
(2) uniformly mixing a bacillus subtilis (CGMCC 1.12938) seed solution (10.0kg) and a substrate (2.0kg) prepared in the step (1), standing and fermenting at 20-25 ℃ for 7 days to obtain a fermented product, centrifuging the fermented product, removing a supernatant, collecting a solid, washing the solid with physiological saline, uniformly mixing the solid with a freeze-drying protective agent (10L) to obtain a suspension, pre-freezing at-70 ℃ for 5h, transferring to a freeze-drying machine, and freeze-drying for 24h to obtain the microecological preparation (hereinafter referred to as a product F).
Example 7 degradation test of pesticide residue in soil
Soil sample:
sample 1: the contents of chlordane and DDT are both 500mg/kg
Sample 2: the content of fenpropathrin and bifenthrin is 500 mg/kg.
The experimental method comprises the following steps: respectively taking 6 parts of 50kg samples 1 and 2, respectively adding products A-F (25 g each), uniformly mixing to ensure that the water content is 20-30%, respectively adding corresponding products A-F (25 g each) after 48 hours, uniformly mixing, and detecting the content of the pesticide in the soil after 48 hours, wherein the results are shown in Table 1.
TABLE 1
Example 8 field experiment
Group T1: product A + rapeseed meal (product A: rapeseed meal ═ 1:5)
Group T2: product B + peanut meal (product B: peanut meal ═ 1:8)
CK1 group: rapeseed dregs
CK2 group: common compound fertilizer (N: P: K ═ 15:15:15)
According to an experimental method, one mu of land is planted with Chinese cabbages in each group, the fertilizing amount is 40 Kg/mu, wherein 40% of the fertilizing amount is used as a base fertilizer, and 60% is used as an additional fertilizer (the additional fertilizer is divided into three times, and each time is 20% of the fertilizing amount); the yield was converted to kg/mu (see Table 2).
TABLE 2
| Experimental group | Yield (kg/mu) |
| T1 group | 11802 |
| T2 group | 11560 |
| CK1 group | 7859 |
| CK2 group | 8656 |
Claims (2)
1. The application of a microecological preparation in degrading pesticide residue is characterized in that the pesticide is selected from organochlorine pesticides or pyrethroid pesticides;
the preparation method of the microecological preparation comprises the following steps:
(1) naturally drying bagasse to constant weight, crushing, soaking in 5-8 wt% cellulase buffer solution at pH of 4.5-6.0 for 20-24 hr, filtering to obtain precipitate, and drying the precipitate to obtain substrate;
(2) uniformly mixing the bacillus subtilis seed solution with the substrate prepared in the step (1), fermenting for 5-7 days at 20-40 ℃ to obtain a fermented product, centrifuging the fermented product, removing a supernatant, collecting a solid, washing the solid with physiological saline, uniformly mixing with a freeze-drying protective agent to obtain a suspension, pre-freezing for 5 hours at-70 ℃, transferring to a freeze dryer, and freeze-drying for 24 hours to obtain the microecological preparation;
the pulverization in the step (1) is selected from pulverization to 40-60 meshes; the buffer solution is selected from citric acid-sodium citrate buffer solution; the enzyme activity of the cellulase is selected from 10-400 KU/g; the mass ratio of the bagasse to the cellulase buffer solution is 1: 8-10;
in the step (2), the mass ratio of the bacillus subtilis seed solution to the substrate is 10: 1-2; 1L of freeze-drying protective agent is used for each kilogram of bacillus subtilis seed liquid; each liter of freeze-drying protective agent contains 90g of glucose, 125g of skim milk powder, 16mL of glycerol and the balance of water; the bacillus subtilis is selected from any one of strains with CGMCC numbers of 1.8955 and 1.8886.
2. Use according to claim 1, characterized in that the pesticide is selected from the group consisting of chlordane, DDT, fenpropathrin, bifenthrin.
Priority Applications (1)
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