CN106957809B - Biological synergistic remediation method for soil organic pollution - Google Patents
Biological synergistic remediation method for soil organic pollution Download PDFInfo
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- 241000982689 Pseudoxanthomonas japonensis Species 0.000 claims description 8
- 230000001737 promoting effect Effects 0.000 claims description 8
- 239000004094 surface-active agent Substances 0.000 claims description 8
- BWDBEAQIHAEVLV-UHFFFAOYSA-N 6-methylheptan-1-ol Chemical compound CC(C)CCCCCO BWDBEAQIHAEVLV-UHFFFAOYSA-N 0.000 claims description 5
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- 229910019142 PO4 Inorganic materials 0.000 claims description 5
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- 239000002054 inoculum Substances 0.000 claims description 4
- 238000012216 screening Methods 0.000 claims description 4
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- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 3
- 229910052708 sodium Inorganic materials 0.000 claims description 3
- 239000011734 sodium Substances 0.000 claims description 3
- RZXLPPRPEOUENN-UHFFFAOYSA-N Chlorfenson Chemical compound C1=CC(Cl)=CC=C1OS(=O)(=O)C1=CC=C(Cl)C=C1 RZXLPPRPEOUENN-UHFFFAOYSA-N 0.000 claims description 2
- CAMHHLOGFDZBBG-UHFFFAOYSA-N epoxidized methyl oleate Natural products CCCCCCCCC1OC1CCCCCCCC(=O)OC CAMHHLOGFDZBBG-UHFFFAOYSA-N 0.000 claims description 2
- HPEUJPJOZXNMSJ-UHFFFAOYSA-N octadecanoic acid methyl ester Natural products CCCCCCCCCCCCCCCCCC(=O)OC HPEUJPJOZXNMSJ-UHFFFAOYSA-N 0.000 claims description 2
- 229940051841 polyoxyethylene ether Drugs 0.000 claims description 2
- 229920000056 polyoxyethylene ether Polymers 0.000 claims description 2
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- 230000012010 growth Effects 0.000 abstract description 10
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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
-
- 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
-
- 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
- B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
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- 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
- C12N1/205—Bacterial isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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Abstract
The invention discloses a microbial agent and a biological synergistic remediation method for soil organic pollution by using the microbial agent, wherein the microbial agent contains effective amount of active ingredients, and the active ingredients comprise Bacillus pumilus (Bacillus pumilus) with the preservation number of CGMCC NO.13208, Gordonia amicalis (Gordonia aminocalilis) with the preservation number of CGMCC NO.4794, Gordonia alkannivorans (Gordonia alkannivorans) with the preservation number of CGMCC NO.4796 and Pseudoxanthomonas japonicus (Pseudomona japonensis) with the preservation number of CGMCC NO. 4797; the present disclosure also provides the application of the microbial agent and a method for treating petroleum pollution by using the microbial agent, wherein the microbial agent can secrete growth hormone to promote the germination and growth of plants, and can inhibit the occurrence of bacterial diseases of the plants by using the microbial agent; the microbial agent and the plants are simultaneously used for treating the petroleum-polluted soil, so that the growth of the soil can be mutually promoted, and the petroleum hydrocarbon can be efficiently decomposed to treat the petroleum pollution.
Description
Technical Field
The disclosure relates to the technical field of organic pollution bioremediation, in particular to a microbial agent and a microbial plant cooperative remediation method for treating soil organic pollution by using the same.
Background
In the world, petroleum is one of the most important energy sources of human beings, and a large amount of petroleum and processed products thereof enter soil to cause petroleum pollution of the soil, so that the petroleum pollution causes harm to human beings and even the whole biosphere, and the petroleum pollution becomes a worldwide environmental problem. Bioremediation is increasingly gaining attention as a potentially efficient, low-cost cleaning technique in the search for petroleum pollution. Bioremediation (Bioremediation) refers to a controlled or spontaneous process of repairing a contaminated environment or eliminating contaminants in an environment by the catalytic degradation of organic contaminants by organisms, in particular microorganisms.
Since 1975, China successively carried out investigation and research on the quantity, distribution, species composition, influence degradation factors and the like of petroleum degrading microorganisms in Bay, Bohai sea, Xiamen, yellow sea and east sea of Qingdao. Among the major factors affecting the degradation of petroleum hydrocarbons by microorganisms, the physiological properties of microorganisms are one of the key factors determining the ability to degrade petroleum hydrocarbons. The adaptability of the microbial strains obtained by separation to environmental conditions is low, and the degradation efficiency of petroleum hydrocarbon in the environment is low, so that the development of a microbial agent capable of efficiently degrading the petroleum hydrocarbon and a biological synergistic remediation method for soil organic pollution is urgently needed.
Disclosure of Invention
The purpose of the present disclosure is to provide a biological synergistic remediation method for organic pollution of soil and microbial agents with strong petroleum hydrocarbon degradation capability, which can effectively treat petroleum pollution.
The present disclosure provides a microbial agent containing an effective amount of an active ingredient comprising Bacillus pumilus (Bacillus pumilus) having a collection number of CGMCC No.13208, Gordonia friendly bacteria (Gordonia aminocalis) having a collection number of CGMCC No.4794, Gordonia alkanivorans (Gordonia alkanivorans) having a collection number of CGMCC No.4796, and Pseudoxanthomonas japonicus (Pseudoxanthomonas japonensis) having a collection number of CGMCC No. 4797.
The disclosure also provides the application of the microbial agent in treating organic pollution.
The present disclosure also provides the use of a microbial agent as described above in promoting plant growth.
The present disclosure also provides a method of treating petroleum pollution comprising growing plants using petroleum-contaminated soil or water while applying a microbial agent as described above.
The disclosure also provides a Bacillus pumilus, wherein the preservation number of the Bacillus pumilus (Bacillus pumilus) is CGMCC NO. 13208.
Through the technical scheme, the synergistic effect among the microbial agents and between the microorganisms and the plants provided by the disclosure is beneficial to efficiently degrading petroleum hydrocarbon organic carbon macromolecules, the microbial agents can secrete growth hormones to promote the germination and growth of the plants, and the microbial agents can inhibit the occurrence of bacterial diseases of the plants; the microbial agent and the plants are simultaneously applied to the treatment of petroleum pollution, so that the growth of the microbial agent and the plants can be mutually promoted, and the microbial agent and the plants are more favorable for efficiently decomposing petroleum hydrocarbon and treating the petroleum pollution.
Additional features and advantages of the disclosure will be set forth in the detailed description which follows.
Biological material preservation
The invention discloses a Bacillus brevis (Bacillus pumilus) which is a pure culture separated from petroleum polluted soil by the inventor of the present disclosure, wherein the preservation number is CGMCC NO.13208, the preservation date is 2016, 10 and 27 days, the preservation unit is the common microorganism center of China Committee for culture Collection of microorganisms, the address is No. 3 Siro No.1 of Xinyang district in Beijing, and the classification name is Bacillus pumilus (Bacillus pumilus). Among them, biological preservation information of Gordonia friendly bacteria (Gordonia aminocalis) with a preservation number of CGMCC No.4794, Gordonia alkaninvorans with a preservation number of CGMCC No.4796, and Pseudoxanthomonas japonicus (Pseudoxanthomonas japonica) with a preservation number of CGMCC No.4797 has been disclosed in patent publication No. CN 102250770A.
Detailed Description
The following describes in detail specific embodiments of the present disclosure. It should be understood that the detailed description and specific examples, while indicating the present disclosure, are given by way of illustration and explanation only, not limitation.
The first aspect of the present disclosure provides a microbial agent, which contains an effective amount of an active ingredient, wherein the active ingredient comprises bacillus pumilus (bacillus pumilus) with a collection number of CGMCC No.13208, Gordonia friendly bacteria (Gordonia aminocalis) with a collection number of CGMCC No.4794, Gordonia alkanivorans (Gordonia alkanivorans) with a collection number of CGMCC No.4796, and Pseudoxanthomonas japonica (Pseudoxanthomonas japonica) with a collection number of CGMCC No. 4797.
Through the technical scheme, the synergistic effect of the microbial agents provided by the disclosure is beneficial to efficiently degrading petroleum hydrocarbon organic carbon macromolecules, the microbial agents can secrete growth hormones to promote the germination and growth of plants, and meanwhile, the microbial agents can inhibit the occurrence of bacterial diseases of the plants; the microbial agent and the plants are simultaneously applied to the treatment of petroleum pollution, so that the growth of the microbial agent and the plants can be mutually promoted, and the microbial agent and the plants are more favorable for efficiently decomposing petroleum hydrocarbon and treating the petroleum pollution.
According to the first aspect of the present disclosure, in order to enable each strain in the microbial agent to act synergistically without inhibiting the growth of other strains, the ratio of the number of bacillus pumilus, gordonia friendly, gordonia alkane-feeding and pseudoxanthomonas japonica is 1: (0.15-7.5): (0.05-5): (0.05-5).
According to the first aspect of the present disclosure, the number of viable bacteria in the microbial agent needs to reach a certain level for the purpose of rapidly exerting the effect of the microbial agent, and the total viable bacteria number is (0.1-2.5) × 10 with respect to 1g of the solid microbial agent or 1mL of the liquid microbial agent10A plurality of; preferably (0.5-1). times.1010And (4) respectively.
According to a first aspect of the present disclosure, the microbial agent further contains a culture medium for maintaining microbial activity; the culture medium can be various common solid culture media or liquid culture media; preferably, the medium is at least one of a beef extract peptone medium, a petroleum screening medium and a PDA medium. For example, the microbial agent can be preserved in a solid beef extract peptone medium; the petroleum screening culture medium is used for screening a microbial agent suitable for petroleum pollution; PDA culture medium can be used for activating and accelerating the growth of the microbial agent; meanwhile, in order to keep the microbial inoculum applied to treating higher activity at the initial stage of petroleum pollution, a liquid beef extract peptone culture medium can be used.
According to the first aspect of the present disclosure, in order to accelerate the decomposition rate of petroleum hydrocarbons by the microbial agent, the microbial agent further contains a surfactant; the content of the surfactant is 0.1-0.5 wt% based on the total weight of the microbial agent.
According to the first aspect of the present disclosure, preferably, the surfactant is at least one of sodium fatty alcohol ether sulfate, sodium methyl stearate polyoxyethylene ether sulfonate, isooctyl alcohol phosphate and isooctyl alcohol ether phosphate.
The second aspect of the present disclosure provides the use of the microbial agent as described above in treating organic pollution, wherein the microbial agent can be used alone in treating petroleum pollution due to its strong petroleum hydrocarbon decomposition capability.
The third aspect of the disclosure provides an application of the microbial agent in promoting plant growth, the microbial agent can be independently applied to promote plant growth, can secrete growth hormone IAA \ GA and the like to promote plant germination and growth, and can inhibit the occurrence of plant bacterial diseases by using the microbial agent.
In a third aspect of the present disclosure, there is provided a method for the bio-synergistic remediation of organic contamination of soil, the method comprising growing plants using organically contaminated soil or water while applying a microbial inoculant as described above; preferably, the organic contamination is petroleum contamination. The microbial agent and the plants are simultaneously applied to the treatment of petroleum pollution, so that the growth of the microbial agent and the plants can be mutually promoted, and the microbial agent and the plants are more favorable for efficiently decomposing petroleum hydrocarbon and treating the petroleum pollution.
The fourth aspect of the disclosure provides a Bacillus pumilus, wherein the preservation number of the Bacillus pumilus (Bacillus pumilus) is CGMCC NO. 13208; the bacillus pumilus is screened from petroleum-polluted soil by the inventor of the present disclosure, and has strong degradation capability on petroleum hydrocarbon.
The present invention will be described in further detail below with reference to examples.
Example 1
Based on the total weight of the strains as 100 percent, inoculating the Bacillus pumilus (Bacillus pumilus) with the preservation number of CGMCC NO.13208 into 100mL of liquid beef extract peptone medium; culturing the inoculated culture medium at 28 deg.C for 1 day at 180 r/min to make the concentration of Bacillus brevis in the culture medium 2.5 × 1010Per mL; the bacillus pumilus agent of the embodiment is prepared by adding 0.1 wt% of sodium fatty alcohol ether sulfate into the culture medium by taking the total weight of the microbial agent as 100%.
Example 2
The preservation number is CGMCC NO.13208 Bacillus pumilus (Bacillus pumilus), Gordonia friendly Gordonia (Gordonia aminocalis) with the preservation number of CGMCC No.4794, Gordonia alkannivorans (Gordonia alkannivorans) with the preservation number of CGMCC No.4796 and Pseudoxanthomonas japonicus (Pseudoxanthomonas japonensis) with the preservation number of CGMCC No.4797 are respectively inoculated in 100mL of liquid beef extract peptone medium; culturing the inoculated culture medium at 28 deg.C for 1 day at 180 r/min to obtain bacterial liquid with concentration of 1 × 109Mixing 9 volume percent of the culture solution containing the bacillus pumilus, 1.5 volume percent of the culture solution containing gordonia friendly Gordonia, 44.75 volume percent of the culture solution containing gordonia alkane-feeding bacteria and 44.75 volume percent of the culture solution containing the pseudomonas japonicus uniformly to prepare 100mL of composite microbial inoculum; the total viable count of 1mL of the microbial agent is 0.1 × 1010And (4) respectively.
Example 3
Respectively inoculating the Bacillus pumilus (Bacillus pumilus) with the preservation number of CGMCC NO.13208, Gordonia friendly bacteria (Gordonia aminocalis) with the preservation number of CGMCC No.4794, Gordonia alkannivorans (Gordonia alkaninvorans) with the preservation number of CGMCC No.4796 and Pseudoxanthomonas japonicus (Pseudoxanthomonas japonensis) with the preservation number of CGMCC No.4797 into 100mL of liquid beef extract peptone medium; culturing the inoculated culture medium at 28 deg.C for 1 day at 180 rpm to obtain bacterial liquid with concentration of 5 × 1010Mixing 5.8 vol% of the culture solution containing the bacillus pumilus, 44 vol% of the culture solution containing the gordonia friendly bacteria, 0.1 vol% of the culture solution containing the gordonia alkane-feeding bacteria, 0.1 vol% of the culture solution containing the pseudomonas japonicus and the balance of a liquid beef extract peptone culture medium uniformly to prepare 100mL of a composite microbial inoculum; the total viable count of 1mL of the microbial inoculum is 2.5 multiplied by 1010And (4) respectively.
Example 4
The Bacillus pumilus (Bacillus pumilus) with the preservation number of CGMCC NO.13208, the Gordonia friendly bacteria (Gordonia aminocalis) with the preservation number of CGMCC NO.4794 and the Gordonia alcalovora (Gor) with the preservation number of CGMCC NO.4796 are mixed to prepare the Bacillus pumilus (Bacillus pumilus) with the preservation number of CGMCC NO.13208, the Gordonia friendly bacteria (Gordonia aminocalis) with the preservation number ofdonia alkannivorans) and Pseudoxanthomonas japonicus (Pseudoxanthomonas japonensis) with the preservation number of CGMCC No.4797 are respectively inoculated in 100mL of liquid beef extract peptone medium; culturing the inoculated culture medium at 28 deg.C for 1 day at 180 r/min to obtain bacterial liquid with concentration of 1 × 1010Mixing 6.25 vol% of the culture solution containing Bacillus pumilus, 18.75 vol% of the culture solution containing Gordonia friendly bacteria, 12.5 vol% of the culture solution containing Gordonia paraffinophilus, 12.5 vol% of the culture solution containing Pseudoxanthomonas japonicus and the balance of liquid beef extract peptone medium uniformly to prepare 100mL of complex microbial inoculum; the total viable count of 1mL of the microbial inoculum is 0.5 multiplied by 1010And (4) respectively.
Example 5
Respectively inoculating the Bacillus pumilus (Bacillus pumilus) with the preservation number of CGMCC NO.13208, Gordonia friendly bacteria (Gordonia aminocalis) with the preservation number of CGMCC No.4794, Gordonia alkannivorans (Gordonia alkaninvorans) with the preservation number of CGMCC No.4796 and Pseudoxanthomonas japonicus (Pseudoxanthomonas japonensis) with the preservation number of CGMCC No.4797 into 100mL of liquid beef extract peptone medium; culturing the inoculated culture medium at 28 deg.C for 1 day at 180 r/min to obtain bacterial liquid with concentration of 1 × 1010Mixing 25 volume percent of the culture solution containing the bacillus pumilus, 25 volume percent of the culture solution containing the gordonia friendly bacteria, 25 volume percent of the culture solution containing the gordonia alkane-feeding bacteria and 25 volume percent of the culture solution containing the pseudomonas japonicus uniformly to prepare 100mL of composite microbial inoculum; the total viable count of 1mL of the microbial agent is 1 × 1010And (4) respectively.
Example 6
Respectively inoculating the Bacillus pumilus (Bacillus pumilus) with the preservation number of CGMCC NO.13208, Gordonia friendly bacteria (Gordonia aminocalis) with the preservation number of CGMCC No.4794, Gordonia alkannivorans (Gordonia alkaninvorans) with the preservation number of CGMCC No.4796 and Pseudoxanthomonas japonicus (Pseudoxanthomonas japonensis) with the preservation number of CGMCC No.4797 into 100mL of liquid beef extract peptone medium; will be provided withThe culture after inoculation is carried out for 1 day under the conditions of 28 ℃ and 180 r/min to ensure that the concentration of the bacterial liquid reaches 1010Mixing 25 volume percent of the culture solution containing the bacillus pumilus, 25 volume percent of the culture solution containing the gordonia friendly bacteria, 25 volume percent of the culture solution containing the gordonia alkane-feeding bacteria and 25 volume percent of the culture solution containing the pseudomonas japonicus uniformly to prepare 100mL of composite microbial inoculum; adding 0.5 wt% of isooctyl alcohol ether phosphate into the composite microbial inoculum by taking the total weight of the microbial inoculum as 100% to prepare the microbial inoculum of the embodiment; the total viable count of 1mL of the microbial agent is 1 × 1010And (4) respectively.
Example 7
Respectively inoculating the Bacillus pumilus (Bacillus pumilus) with the preservation number of CGMCC NO.13208, Gordonia friendly bacteria (Gordonia aminocalis) with the preservation number of CGMCC No.4794, Gordonia alkannivorans (Gordonia alkaninvorans) with the preservation number of CGMCC No.4796 and Pseudoxanthomonas japonicus (Pseudoxanthomonas japonensis) with the preservation number of CGMCC No.4797 into 100mL of liquid beef extract peptone medium; culturing the inoculated culture medium at 28 ℃ for 1 day at 180 r/min until the bacterial liquid concentration reaches 1010More than one strain per mL, 6.25 volume percent of the culture solution containing the bacillus pumilus, 18.75 volume percent of the culture solution containing the gordonia bacteria, 12.5 volume percent of the culture solution containing the gordonia alkane-feeding bacteria, 12.5 volume percent of the culture solution containing the pseudomonas japonicus and the balance of liquid beef extract peptone culture medium are uniformly mixed to prepare 100mL of composite microbial inoculum; adding 0.1 wt% of fatty alcohol ether sodium sulfate into the complex microbial inoculum by taking the total weight of the microbial inoculum as 100% to prepare the microbial inoculum of the embodiment; the total viable count of 1mL of the microbial inoculum is 0.5 multiplied by 1010And (4) respectively.
Comparative example 1
A microbial agent was prepared according to the method of example 1, except that the Bacillus pumilus having the accession number of CGMCC NO.13208 was replaced with the Bacillus pumilus having the accession number of PDBP-2, which was isolated by the inventors.
Comparative example 2
A microbial agent was prepared according to the method of example 3, except that the Bacillus pumilus having the accession number of CGMCC NO.13208 was replaced with the Bacillus pumilus having the accession number of PDBP-3, which was isolated by the inventors.
Test example 1
The test examples were conducted to determine the efficiency of the microbial agents of examples 1-7 and comparative examples 1-2 in decomposing petroleum hydrocarbons.
Weighing a petroleum-polluted soil sample, wherein each part is 10g (the petroleum content is 1.25g), and the soil sample is obtained by mixing the soil polluted by the fallen crude oil, the oily sediment and the plough layer soil stacked with the oily sediment for a long time around a production well of a Clarity oil field according to the ratio of 1: 1: 1, mixing the petroleum polluted soil sample according to the proportion of the mixture; adding the petroleum-contaminated soil sample into 250mL triangular flasks filled with 100mL liquid beef extract peptone respectively, and simultaneously inoculating 3 bottles of the microorganisms in the examples 1-7, the comparative example 1 or the comparative example 2 respectively; after inoculation, the culture medium is cultured for 5 days under the conditions of 28 ℃ and 180 r/min, and the petroleum content (g) in the culture medium is detected, and specific results are shown in a table 1.
TABLE 1
As can be seen from comparison of examples 1-7 and comparative examples 1-2 in Table 1, the microbial agent containing Bacillus pumilus with the preservation number of CGMCCNo.13208 provided by the present disclosure has better petroleum hydrocarbon decomposition effect; the complex microbial agent containing various strains in examples 2-5 has higher petroleum hydrocarbon decomposition efficiency than that of example 1; more preferably, the number of viable bacteria in the complex microbial agent described in examples 4 to 7 is (0.5 to 1.0). times.1010The petroleum hydrocarbon decomposition efficiency is higher when the amount per mL is larger; most preferably, the microbial agents of examples 6-7 have better petroleum hydrocarbon decomposition effect when 0.1-0.5 wt% of surfactant is added.
Test example 2
The test examples were used to determine the efficiency of the microbial agents of examples 1-7 and comparative examples 1-2 in promoting plant growth.
100 alfalfa seeds are taken and put into a culture dish with the thickness of 100mm, the culture dish is paved with 3 pieces of filter paper, the diluted alfalfa seeds are treated by a microbial agent with the content of 10 volume percent, the filter paper is kept soaked by using sterile water as a control, the germination potential of the seeds is measured after the seeds are cultured for 3 days at room temperature, the germination rate and the root length are measured after 5 days, and specific results are shown in table 2.
TABLE 2
As can be seen from comparison between examples 1-7 and comparative examples 1-2 in Table 2, the microbial agent containing Bacillus pumilus with the preservation number of CGMCCNo.13208 provided by the present disclosure has better effect of promoting plant growth; the complex microbial agents of examples 2 to 7 containing a plurality of strains are more effective than those of example 1, and preferably, the number of viable bacteria in the complex microbial agents of examples 4 to 7 is (0.5 to 1.0). times.1010The effect of promoting the growth of the plants is better when the plant growth is one/mL.
Test example 3
The test example shows the effect of the microbial agent in examples 1-7 and comparative examples 1-2 in combination with plants on treating petroleum-contaminated soil.
Weighing 100g (oil content is 5g) of oil-contaminated soil samples, wherein the oil-contaminated soil samples are 100g (oil content is 5g) of the oil-contaminated soil samples, namely, the soil samples are obtained by the method that the oil wells of the Clarity oil field are filled with the fallen crude oil contaminated soil, the oily sediment and the plough layer soil with the oily sediment stacked for a long time according to the weight ratio of 1: 1: 1, mixing the petroleum polluted soil sample according to the proportion of the mixture; spreading the petroleum-polluted soil sample in a culture dish with the thickness of 100mm, taking 100 alfalfa seeds, respectively putting the alfalfa seeds in the culture dish paved with the oil-polluted soil, irrigating with 10 volume percent of microbial inoculum, and taking sterile water as a reference; the petroleum-contaminated soil was kept infiltrated and cultured at room temperature for 5 days, then the germination potential of the seeds was measured, and after 10 days, the germination percentage, root length and petroleum decomposition rate of alfalfa were measured. The specific results are shown in Table 3.
TABLE 3
As can be seen from comparison between examples 1-7 and comparative examples 1-2 in Table 3, the microbial agent containing Bacillus pumilus with the preservation number of CGMCCNo.13208, provided by the disclosure, has a better effect of treating petroleum pollution in combination with plants, and is beneficial to the growth of the plants; the compound microbial agent containing various strains in the examples 2-7 has better effects of promoting the growth of plants and decomposing oil stains compared with the compound microbial agent in the example 1; more preferably, the number of viable bacteria in the complex microbial agent described in examples 4 to 7 is (0.5 to 1.0). times.1010The effect of promoting the growth of plants and decomposing oil stains is better when the plants are planted per mL; most preferably, the microbial agents described in examples 6-7 have the best petroleum decomposition effect when 0.1-0.5 wt% of a surfactant is added thereto.
The preferred embodiments of the present disclosure have been described in detail above, however, the present disclosure is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present disclosure within the technical idea of the present disclosure, and these simple modifications all fall within the protection scope of the present disclosure.
It should be noted that the various features described in the above embodiments may be combined in any suitable manner without departing from the scope of the invention. In order to avoid unnecessary repetition, various possible combinations will not be separately described in this disclosure.
In addition, any combination of various embodiments of the present disclosure may be made, and the same should be considered as the disclosure of the present disclosure, as long as it does not depart from the spirit of the present disclosure.
Claims (10)
1. The microbial agent is characterized by containing an effective amount of active ingredients, wherein the active ingredients comprise Bacillus pumilus (Bacillus pumilus) with the preservation number of CGMCC NO.13208, Gordonia friendly bacteria (Gordonia aminocalis) with the preservation number of CGMCC NO.4794, Gordonia alkane-feeding bacteria (Gordonia alkanivorans) with the preservation number of CGMCC NO.4796 and CGMCC No4797, (Pseudoxanthomonas japonicus (Pseudoxanthomonas japonensis); the number ratio of the bacillus pumilus, the gordonia friendly gordonia, the gordonia alkane-feeding gordonia and the pseudoxanthomonas japonica is 1: (0.15-7.5): (0.05-5): (0.05-5); the total viable count is (0.1-2.5) x 10 relative to 1g of microbial agent or 1mL of liquid microbial agent10And (4) respectively.
2. The microbial agent according to claim 1, wherein the total viable cell count is (0.5 to 1). times.10 relative to 1g of the microbial agent or 1mL of the liquid microbial agent10And (4) respectively.
3. The microbial agent according to claim 1, wherein the microbial agent further comprises a medium for maintaining microbial activity.
4. The microbial agent according to claim 3, wherein the medium is at least one of beef extract peptone medium, petroleum screening medium and PDA medium.
5. The microbial agent of claim 1, wherein the microbial agent further comprises a surfactant; the content of the surfactant is 0.1-0.5 wt% based on the total weight of the microbial agent.
6. The microbial agent according to claim 5, wherein the surfactant is at least one of sodium fatty alcohol ether sulfate, sodium methyl stearate polyoxyethylene ether sulfonate, isooctyl alcohol phosphate and isooctyl alcohol ether phosphate.
7. Use of the microbial inoculant of any one of claims 1 to 6 for the remediation of petroleum pollution.
8. Use of the microbial inoculant according to any one of claims 1 to 6 for promoting plant growth.
9. A method for the bio-synergistic remediation of organic contaminated soil, the method comprising growing plants in petroleum contaminated soil or water and applying the microbial inoculant of any one of claims 1 to 6.
10. The Bacillus pumilus is characterized in that the preservation number of the Bacillus pumilus (Bacillus pumilus) is CGMCC NO. 13208.
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| CN107803400B (en) * | 2017-10-31 | 2020-12-08 | 中国环境科学研究院 | Composting method for restoring petroleum hydrocarbon polluted soil by utilizing biogas slurry |
| CN112159060B (en) * | 2020-09-29 | 2022-05-17 | 重庆大学 | Microorganism-combined plant-type composite deodorant for black-odor rivers and preparation method thereof |
| CN112400546B (en) * | 2021-01-22 | 2021-04-27 | 中国农业科学院农业环境与可持续发展研究所 | Ecological rod and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101717725A (en) * | 2009-12-25 | 2010-06-02 | 中国海洋石油总公司 | Microbial inoculum for bioremediating coastline polluted by oil spilling and preparation method thereof |
| CN102250770A (en) * | 2011-06-22 | 2011-11-23 | 中国农业科学院农业环境与可持续发展研究所 | Microbial agent and preparation method and application thereof |
| CN102250801A (en) * | 2011-06-22 | 2011-11-23 | 中国农业科学院农业环境与可持续发展研究所 | Microbial agent, and its preparation method and application |
| CN102839137A (en) * | 2012-07-27 | 2012-12-26 | 长安大学 | Solid microbial bacterium agent for petroleum degradation and preparation method thereof |
| CN105316268A (en) * | 2015-12-08 | 2016-02-10 | 山东宝源生物有限公司 | Bacillus pumilus strain for producing gibberellin and application of bacillus pumilus strain in petroleum degradation |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101717725A (en) * | 2009-12-25 | 2010-06-02 | 中国海洋石油总公司 | Microbial inoculum for bioremediating coastline polluted by oil spilling and preparation method thereof |
| CN102250770A (en) * | 2011-06-22 | 2011-11-23 | 中国农业科学院农业环境与可持续发展研究所 | Microbial agent and preparation method and application thereof |
| CN102250801A (en) * | 2011-06-22 | 2011-11-23 | 中国农业科学院农业环境与可持续发展研究所 | Microbial agent, and its preparation method and application |
| CN102839137A (en) * | 2012-07-27 | 2012-12-26 | 长安大学 | Solid microbial bacterium agent for petroleum degradation and preparation method thereof |
| CN105316268A (en) * | 2015-12-08 | 2016-02-10 | 山东宝源生物有限公司 | Bacillus pumilus strain for producing gibberellin and application of bacillus pumilus strain in petroleum degradation |
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