CN112457862A - Biological soil conditioner and application thereof - Google Patents
Biological soil conditioner and application thereof Download PDFInfo
- Publication number
- CN112457862A CN112457862A CN202011417963.3A CN202011417963A CN112457862A CN 112457862 A CN112457862 A CN 112457862A CN 202011417963 A CN202011417963 A CN 202011417963A CN 112457862 A CN112457862 A CN 112457862A
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- Prior art keywords
- bacillus subtilis
- soil conditioner
- biological soil
- bacillus amyloliquefaciens
- bacillus
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/40—Soil-conditioning materials or soil-stabilising materials containing mixtures of inorganic and organic compounds
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/08—Reclamation of contaminated soil chemically
-
- 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
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2101/00—Agricultural use
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2109/00—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE pH regulation
Abstract
The invention discloses a biological soil conditioner and application thereof, wherein the biological soil conditioner comprises the following raw materials in percentage by weight: 21 to 23 percent of turf; 26-30% of mineral source potassium fulvate; 26-30% of fermented straw powder; 19 to 20 percent of bran coat; 1-3% of microbial agent; wherein the microbial agent consists of a Bacillus subtilis LX13 zymocyte liquid, a Bacillus amyloliquefaciens zymocyte liquid and a carrier; the Bacillus subtilis LX13 is preserved in the China general microbiological culture Collection center of the culture Collection of microorganisms with the preservation number of CGMCC No. 8805; the bacillus amyloliquefaciens is preserved in China center for type culture Collection with the preservation number of CCTCC: AB 2014337. The invention can effectively treat the soil with heavy metals exceeding the standard.
Description
Technical Field
The invention relates to the field of soil improvement preparations. More specifically, the invention relates to a biological soil conditioner and application thereof.
Background
Environmental protection is an inevitable requirement for global economic sustainable development, and the awareness of environmental protection of people is gradually increased. At present, various metallurgical plants and chemical plants are built around rural areas, discharge a large amount of toxic and harmful substances, cause irreversible serious pollution to local soil, change the chemical components of the soil, cause the content of heavy metals in the soil to exceed the standard, and cause serious threats to the health of surrounding crops and surrounding masses. The treatment of soil with excessive heavy metals becomes an important task for current environmental protection.
At present, the soil with over-standard heavy metal content is mainly treated by means of physical control, biological control, chemical control or comprehensive control, but the treatment is not ideal.
Disclosure of Invention
The invention aims to provide a biological soil conditioner and application thereof, which can effectively treat soil with excessive heavy metals.
To achieve these objects and other advantages in accordance with the present invention, there is provided a biological soil amendment, comprising the following raw materials in weight percent:
wherein the microbial agent consists of a Bacillus subtilis LX13 zymocyte liquid, a Bacillus amyloliquefaciens zymocyte liquid and a carrier; the Bacillus subtilis LX13 is preserved in the China general microbiological culture Collection center of the culture Collection of microorganisms with the preservation number of CGMCC No. 8805; the bacillus amyloliquefaciens is preserved in China center for type culture Collection with the preservation number of CCTCC: AB 2014337.
Preferably, the carrier is one or a combination of two of bean pulp and cassava residue.
Preferably, the effective viable count of the Bacillus subtilis LX13 in the Bacillus subtilis LX13 zymocyte liquid is 2.0 multiplied by 1010～2.5×1011cfu/mL；
The effective viable count of the bacillus amyloliquefaciens in the bacillus amyloliquefaciens zymophyte liquid is 3.0 multiplied by 1010～1×1011cfu/mL；
The total volume of the Bacillus amyloliquefaciens fermented liquid and the Bacillus subtilis LX13 fermented liquid and the total weight of the carrier are 1 mL: 5-6 g.
Preferably, the particle size of the carrier is 30 mesh.
Preferably, the Bacillus subtilis LX13 is inoculated in an LB culture medium and subjected to shake flask fermentation culture to obtain a Bacillus subtilis LX13 fermented bacterial liquid, wherein the culture temperature is 29 ℃, and the Bacillus subtilis LX13 fermented bacterial liquid is cultured for 2-3 days;
and inoculating the bacillus amyloliquefaciens into an LB culture medium, and performing shake-flask fermentation culture to obtain the bacillus amyloliquefaciens zymocyte liquid, wherein the culture temperature is 30 ℃, and the culture is performed for 2-3 days.
The invention also provides application of the biological soil conditioner in repairing heavy metal contaminated soil.
Preferably, the application amount of the biological soil conditioner is 230-300 kg/mu.
The invention at least comprises the following beneficial effects: the mineral source potassium fulvate in the biological soil conditioner is an alkaline substance, and the pH value of soil can be adjusted; the grass carbon, the fermented straw powder and the bran coat have the function of adsorbing and passivating heavy metals; the synergistic effect of the two zymocyte liquids in the microbial agent can influence the adsorption and passivation effect of the passivator on Cd, reduce the accumulation of Cd on a seed part, and ensure the safety of rice planting.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Detailed Description
The present invention is further described in detail below with reference to examples so that those skilled in the art can practice the invention with reference to the description.
It is to be noted that the experimental methods described in the following embodiments are all conventional methods unless otherwise specified, and the reagents and materials are commercially available unless otherwise specified.
The invention provides a biological soil conditioner, which comprises the following raw materials in percentage by weight:
the microbial agent is prepared by mixing Bacillus subtilis LX13 zymocyte liquid, Bacillus amyloliquefaciens zymocyte liquid and a carrier. The Bacillus subtilis LX13 is preserved in the China general microbiological culture Collection center of the culture Collection of microorganisms with the preservation number of CGMCC No. 8805; the bacillus amyloliquefaciens is preserved in China center for type culture Collection with the preservation number of CCTCC: AB 2014337.
In the invention, the carrier is one or a combination of soybean meal and cassava residue, and more preferably the mixture of the soybean meal and the cassava residue in a mass ratio of 1-3: 1, and preferably, the carrier is crushed and then sieved by a 30-mesh sieve. The vector is sterilized at 125 ℃, and then mixed with Bacillus subtilis LX13 zymocyte liquid and Bacillus amyloliquefaciens zymocyte liquid.
In the invention, the effective viable count of the Bacillus subtilis LX13 in the Bacillus subtilis LX13 fermented liquid is 2.0 multiplied by 1010～2.5×1011cfu/mL。
The effective viable count of the bacillus amyloliquefaciens in the bacillus amyloliquefaciens zymophyte liquid is 3.0 multiplied by 1010～1×1011cfu/mL。
The total volume of the Bacillus amyloliquefaciens fermented liquid and the Bacillus subtilis LX13 fermented liquid and the total weight of the carrier are 1 mL: 5-6 g.
In the invention, the Bacillus subtilis LX13 is inoculated in an LB solid culture medium and subjected to shake flask fermentation culture to obtain a Bacillus subtilis LX13 fermented bacterial liquid, the culture temperature is 29 ℃, and the Bacillus subtilis LX13 fermented bacterial liquid is cultured for 2-3 days.
And inoculating the bacillus amyloliquefaciens into an LB solid culture medium, and performing shake flask fermentation culture to obtain the bacillus amyloliquefaciens zymocyte liquid, wherein the culture temperature is 30 ℃, and the culture is performed for 2-3 days.
The LB solid culture medium comprises the following raw materials: the preparation amount is 1L, tryptone is 10g, yeast extract is 5g, sodium chloride is 10g, agar is 15g, and the balance is water.
The invention also provides application of the biological soil conditioner in repairing heavy metal contaminated soil, wherein the application amount of the biological soil conditioner is 230-300 kg/mu.
Example 1
The preparation of the biological soil conditioner comprises the following steps:
1. preparation of microbial agent
(1) Preparation of a Bacillus subtilis LX13 zymocyte liquid: inoculating Bacillus subtilis LX13 into an LB solid culture medium, and performing shake flask fermentation culture to obtain a Bacillus subtilis LX13 fermented liquid, wherein the culture temperature is 29 ℃, and the Bacillus subtilis LX13 fermented liquid is cultured for 3 days.
Preparing a bacillus amyloliquefaciens zymocyte liquid: inoculating the bacillus amyloliquefaciens into an LB solid culture medium, and performing shake flask fermentation culture to obtain the bacillus amyloliquefaciens zymocyte liquid, wherein the culture temperature is 30 ℃, and the culture is performed for 3 days.
The LB solid culture medium comprises the following raw materials: the preparation amount is 1L, tryptone is 10g, yeast extract is 5g, sodium chloride is 10g, agar is 15g, and the balance is water.
Preparation of the carrier: mixing bean pulp and cassava dregs according to a mass ratio of 2: 1, mixing, crushing and sieving by a 30-mesh sieve.
(2) According to the total volume of the Bacillus amyloliquefaciens zymocyte liquid and the Bacillus subtilis LX13 zymocyte liquid and the total weight of the carrier being 1 mL: 5g of mixture, wherein the volume ratio of the Bacillus amyloliquefaciens fermented bacterial liquid to the Bacillus subtilis LX13 fermented bacterial liquid is 1: 1.
2. preparation of biological soil conditioner
The fertilizer is prepared by mixing 23% of turf, 28% of mineral potassium fulvate, 28% of fermented straw powder, 19% of bran coat and 2% of microbial agent in percentage by weight, and the pH value is 7.48.
Example 2
The biological soil conditioner is prepared by mixing 21% of turf, 26% of mineral potassium fulvate, 30% of fermented straw powder, 20% of bran and 3% of microbial agent in percentage by weight. The microbial agent is prepared from Bacillus amyloliquefaciens zymocyte liquid, Bacillus subtilis LX13 zymocyte liquid and carrier bean pulp according to the proportion of 0.4 mL: 0.5 mL: 6g of the above-mentioned materials are mixed.
Comparative example 1
A biological soil conditioner is prepared by mixing 23% of turf, 28% of mineral potassium fulvate, 28% of fermented straw powder, 19% of bran and 2% of microbial agent in percentage by weight, wherein the pH value is 7.53. Wherein the microbial agent is Bacillus subtilis LX13 zymocyte liquid and the total weight of the carrier is 1 mL: 5g of the components are mixed.
Preparation of a Bacillus subtilis LX13 zymocyte liquid: inoculating Bacillus subtilis LX13 into an LB solid culture medium, and performing shake flask fermentation culture to obtain a Bacillus subtilis LX13 fermented liquid, wherein the culture temperature is 29 ℃, and the Bacillus subtilis LX13 fermented liquid is cultured for 3 days. The LB solid culture medium comprises the following raw materials: the preparation amount is 1L, tryptone is 10g, yeast extract is 5g, sodium chloride is 10g, agar is 15g, and the balance is water.
Preparation of the carrier: mixing bean pulp and cassava dregs according to a mass ratio of 2: 1, mixing, crushing and sieving by a 30-mesh sieve.
Comparative example 2
A biological soil conditioner is prepared by mixing 23% of turf, 28% of mineral potassium fulvate, 28% of fermented straw powder, 19% of bran and 2% of microbial agent in percentage by weight, wherein the pH value is 7.56. Wherein the microbial agent is bacillus amyloliquefaciens zymocyte liquid volume and the total weight of the carrier is 1 mL: 5g of the components are mixed.
Preparing a bacillus amyloliquefaciens zymocyte liquid: inoculating the bacillus amyloliquefaciens into an LB solid culture medium, and performing shake flask fermentation culture to obtain the bacillus amyloliquefaciens zymocyte liquid, wherein the culture temperature is 30 ℃, and the culture is performed for 3 days. The LB solid culture medium comprises the following raw materials: the preparation amount is 1L, tryptone is 10g, yeast extract is 5g, sodium chloride is 10g, agar is 15g, and the balance is water.
Preparation of the carrier: mixing bean pulp and cassava dregs according to a mass ratio of 2: 1, mixing, crushing and sieving by a 30-mesh sieve.
Field test
1) Design of experiments
The soil around a certain smelting factory in Ezhou city has pollution of different degrees, and the polluted farmland is repaired by using the biological soil conditioner. And (4) ploughing and uniformly raking 0-20 cm of soil on the surface layer of the farmland where early rice needs to be planted at the end of 2018 in 4 months, and then sampling for detection.
At the end of 4 months in the year, the biological soil conditioner prepared in the embodiment 1 (group A), the comparative example 1 (group B) and the comparative example 2 (group C) is respectively and uniformly applied to correspondingly divided farmland areas, the application amount is 230 kg/mu, the specification of the farmland areas is 7m by 7m, the farmland areas are separated by plastic impermeable films, a group of blank control groups (group D) is simultaneously designed, 4 groups are counted, each group is repeated for three times, the average value is taken, conventional base fertilizer (N/P/K fertilizer) is applied to the farmland in the middle and ten days of May, the transplanting variety is 287 early rice seedlings, the transplanting row spacing is 30cm by 20cm, the conventional base fertilizer is supplemented in each growth period of the rice, the rice is harvested in the lower ten days of 7 months, and 10 rice and soil samples located at 0-3 cm of the root system of the rice are randomly collected in each farmland area.
2) Measurement method
After rice is collected, the rice is washed for three times by ultrapure water, then is dried at the temperature of 60 ℃, and the seed part of the rice is separated to be used as a rice sample, crushed and dried for later use.
The pH value of the soil is measured by a potential method, and the content of Organic Matters (OM) in the soil and the Cation Exchange Capacity (CEC) of the soil are measured by a soil agricultural chemical analysis method; the method comprises the following steps of (1) digesting a rice sample by a digestion method in the determination of the Cd content of the rice sample, and determining by an atomic absorption spectrometry; the method for measuring the content of Cd in the soil active state adopts 'determination of DTPA/ICP atomic emission spectrometry' of soil active state elements.
3) Test results
The results of physicochemical properties of the soil samples before 4-month end treatment in 2018 are shown in table 1.
TABLE 1
The results of physicochemical properties of the soil samples in late 7 th month of 2018 are shown in Table 2.
TABLE 2
The Cd absorption accumulation amount of the rice sample in late 7 months in 2018 is shown in Table 3.
TABLE 3
Through table 1 and table 2 contrast, can know, the biological soil amendment of this application has reconciled the problem of soil faintly acid, can effectively reduce the available state Cd content in the soil, improves CEC content. On one hand, the mineral potassium fulvate is an alkaline substance and can adjust the pH value of soil, and the turf, the fermented straw powder and the bran coat are used as passivators and have an adsorption and passivation effect on heavy metals; on the other hand, the synergistic effect of the two zymocyte liquids in the microbial agent is better than that of a single strain, and the adsorption and passivation effects of the passivator on Cd can be influenced. As can be seen from Table 3, after the biological soil conditioner disclosed by the application is used, the absorption and accumulation of heavy metal Cd by grains are reduced by 79.66%, and the effect is obviously better than that of the group B and the group C. The biological soil conditioner of the group A can obviously reduce the accumulation of Cd in a seed part, and the safety of rice planting is guaranteed.
While embodiments of the invention have been described above, it is not limited to the applications set forth in the description and the embodiments, which are fully applicable to various fields of endeavor for which the invention may be embodied with additional modifications as would be readily apparent to those skilled in the art, and the invention is therefore not limited to the details given herein and to the embodiments shown and described without departing from the generic concept as defined by the claims and their equivalents.
Claims (7)
1. The biological soil conditioner is characterized by comprising the following raw materials in percentage by weight:
wherein the microbial agent consists of a Bacillus subtilis LX13 zymocyte liquid, a Bacillus amyloliquefaciens zymocyte liquid and a carrier; the Bacillus subtilis LX13 is preserved in the China general microbiological culture Collection center of the culture Collection of microorganisms with the preservation number of CGMCC No. 8805; the bacillus amyloliquefaciens is preserved in China center for type culture Collection with the preservation number of CCTCC: AB 2014337.
2. The biological soil conditioner of claim 1, wherein said carrier is one or a combination of soybean meal and cassava dregs.
3. The biological soil conditioner according to claim 2, wherein the effective viable count of Bacillus subtilis LX13 in the Bacillus subtilis LX13 fermented liquid is 2.0 x 1010～2.5×1011cfu/mL；
The effective viable count of the bacillus amyloliquefaciens in the bacillus amyloliquefaciens zymophyte liquid is 3.0 multiplied by 1010～1×1011cfu/mL；
The total volume of the Bacillus amyloliquefaciens fermented liquid and the Bacillus subtilis LX13 fermented liquid and the total weight of the carrier are 1 mL: 5-6 g.
4. The biological soil conditioner of claim 2, wherein said carrier has a particle size of 30 mesh.
5. The biological soil conditioner according to claim 1, wherein the Bacillus subtilis LX13 is inoculated in an LB culture medium and cultured in a shake flask fermentation way to obtain a Bacillus subtilis LX13 zymocyte liquid, the culture temperature is 29 ℃, and the Bacillus subtilis LX13 zymocyte liquid is cultured for 2-3 days;
and inoculating the bacillus amyloliquefaciens into an LB culture medium, and performing shake-flask fermentation culture to obtain the bacillus amyloliquefaciens zymocyte liquid, wherein the culture temperature is 30 ℃, and the culture is performed for 2-3 days.
6. Use of the biological soil conditioner of any one of claims 1 to 5 for remediation of heavy metal contaminated soil.
7. The use of claim 6, wherein the amount of biological soil amendment applied is 230 to 300 kg/acre.
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| CN202011417963.3A CN112457862A (en) | 2020-12-07 | 2020-12-07 | Biological soil conditioner and application thereof |
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| CN202011417963.3A CN112457862A (en) | 2020-12-07 | 2020-12-07 | Biological soil conditioner and application thereof |
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| CN117402767A (en) * | 2023-02-01 | 2024-01-16 | 齐鲁工业大学(山东省科学院) | Composite microbial agent and application thereof in crop planting and soil improvement |
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