CN104772328B - A kind of biological prosthetic oil hydrocarbon compound contaminated soil method and application - Google Patents

A kind of biological prosthetic oil hydrocarbon compound contaminated soil method and application Download PDF

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CN104772328B
CN104772328B CN201510154576.8A CN201510154576A CN104772328B CN 104772328 B CN104772328 B CN 104772328B CN 201510154576 A CN201510154576 A CN 201510154576A CN 104772328 B CN104772328 B CN 104772328B
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soil
biological agent
method described
plant
plant growth
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CN104772328A (en
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黄晓东
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Evergrande Oasis Qingdao Ecological Prataculture Co ltd
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Abstract

The present invention provides a kind of biological prosthetic oil hydrocarbon compound contaminated soil methods, the method includes planting rehabilitation plant in the soil and applying soil to repair biological agent, the rehabilitation plant is one or more combinations in annual ryegrass, English ryegrass, tall grass and grassland gramineous crop;The reparation biological agent includes at least the bacterium solution containing plant growth-promoting bacteria, and the bacterium solution contains plant growth-promoting bacteria Pseudomonas cedrina UW3 and Pseudomonas putida UW4.The method of the present invention can greatly shorten the time of soil remediation, usually in a seriously polluted area, can control the level repaired in 35 years to before pollution.

Description

A kind of biological prosthetic oil hydrocarbon compound contaminated soil method and application
Technical field
The present invention relates to soil remediation fields, specifically, are related to a kind of biological prosthetic oil hydrocarbon compound contaminated soil Method and application.
Background technology
With the development of modernization industry, soil pollution has been increasingly becoming one of global environmental problem.Especially dig up mine, Metal smelt, oil exploitation and Production in Chemical Plant are the most serious to soil pollution.Cause heavy metal, hydrocarbon compound and each Toxic salt too high levels in the soil are planted, and are difficult to eliminate by natural decomposition for a long period of time clean.
Containing a large amount of harmful substances in petroleum hydrocarbon, such as benzene homologues, polycyclic aromatic hydrocarbon and other are difficult to degrade toxic Harmful substance.Toxic action is not only directly generated to human body by petroleum hydrocarbon contaminated soil, also results in grain drop in production.
Petroleum hydrocarbon hydrophobicity is strong, it is difficult to be desorbed from soil, rate of release is slow in soil matrix, is persistence Pollution sources.In addition, this slower mass transport velocity, constrains the degradation to pollutant such as microorganism, soil is given The application of recovery technique brings difficulty.
The oil gas field more than totally 400 of China's exploration, exploitation, is distributed in national 25 provinces, municipalities and autonomous regions, field area work Nearly 200,000 square kilometres of range.According to incompletely statistics, in field area there is at more than 20 ten thousand contaminated site, high concentration petroleum pollution soil And oil sediment burden exceedes 2,000,000 tons.Many farmlands around North China Oilfield can not be cultivated due to crude oil pollution, every year will Substantial contribution is paid as the reparation to peasant;Yellow River System annual oil-containing reaches as high as 4.82mg/L, and Liaohe River water system year is flat Equal oil-containing reaches as high as 7.68mg/L, it will be apparent that has been more than the standard (< 0.1mg/L) of the national three-level surface water;Some areas are very It is more than national standard if In Zibo, Shandong underground water highest oil-containing has reached 100mg/L or more to groundwater resources are polluteed 1000 times of (< 0.1mg/L) or more.
The petroleum hydrocarbon contaminated problem for how eliminating soil, has become very urgent problem.
Invention content
In order to solve above-mentioned prior art problem, it is an object of the present invention to provide a kind of biological prosthetic oil hydrocarbonylations Object contaminated soil method is closed, the method includes planting rehabilitation plant in the soil and apply soil to repair biology system Agent.
Another object of the present invention is to provide the rehabilitation plants and reparation biological agent to combine in soil remediation Using.
In order to achieve the above object, on the one hand, the present invention provides a kind of biological prosthetic oil hydrocarbon compound contaminated soil sides Method, the method includes planting rehabilitation plant in the soil and applying soil to repair biological agent, the rehabilitation plant is Annual ryegrass (Lolium multiflorum), English ryegrass (Lolium perenne), tall grass One or more combinations in (Festuca arundinacea) and grassland gramineous crop;The reparation biological agent is extremely Include the bacterium solution containing plant growth-promoting bacteria less, the bacterium solution contain plant growth-promoting bacteria Pseudomonas cedrina UW3 and Pseudomonas putida UW4。
Plant growth-promoting bacteria can inhibit the generation of ethylene in plant, and synthesize beneficial auxin, to promote plant to give birth to It is long.And it is a discovery of the invention that plant growth-promoting bacteria can also promote oil hydrocarbon compound pollute degradation and root system of plant to pollution The absorption of object, it might even be possible to protect the plants from the injury of pollutant in soil.
Further, the present invention through a large number of experiments, it is found that it is good selected strain and specific plant have Synergistic effect, the oil hydrocarbon compound removed in soil that can be more efficient pollute.
The wherein present invention preferably soil is by oil hydrocarbon compound contaminated soil.
According to embodiment preferred for this invention, wherein it is preferred that grassland grass family effect includes rye or small black Wheat.
Wherein annual ryegrass (Lolium multiflorum) of the invention, English ryegrass (Lolium Perenne), tall grass (Festuca arundinacea) and grassland gramineous crop are commercially available, are for example purchased from www.aerc.com;
Pseudomonas cedrina UW3 and Pseudomonas putida UW4 therein are recorded in Chinese patent Shen Please be in 200810181327.8, the present invention no longer repeats.
For example, in bacterium solution, the bacterium number ratio of UW3 and UW4 are 1:2~2:1.
Again for example, the reparation biological agent further includes composite carrier, the composite carrier include organic matter, Two or more arbitrary combination in humic acid, decomposed organic matter, complexing displacer, embedded material;It is preferred that complex carrier The ratio of material and bacterium solution is at least 0.2~200,000,000/every gram composite carrier of bacterium number of plant growth-promoting bacteria;Further preferably with compound On the basis of the weight of carrier material is 100%, the composite carrier includes the ingredient of following weight percent:15~ 35% organic matter, 12~32% humic acid, 12~32% decomposed organic matter, 5~22% complexing displacer and 0.01 ~1% embedded material.
According to embodiment preferred for this invention, wherein the method further includes the pre-treatment step to soil.
The pre-treatment step volume can be conventional for this field pre-treatment step, it is currently preferred to be:It is described pre- Processing step include prepartion of land, except stone, plough, rake the soil level, apply fertilizer and sow before preparation one or more of.
According to embodiment preferred for this invention, wherein the method further include after planting rehabilitation plant to soil into Row irrigates, cuts off and apply fertilizer.
According to another preferred embodiment of the present invention, wherein the method further includes by cut substrate after cutting off from soil Middle removing.
According to embodiment preferred for this invention, wherein described cut off as in the 60-70 of rehabilitation plant growth course It cuts off for the first time, and later primary per cutting off within 40-60 days, cuts off rear toothing 10-12cm every time.
Wherein more preferably cut off in the 60th day first time.
According to embodiment preferred for this invention, wherein the cutting can be according to above-mentioned cutting interval through plant Entire growth period;Soil sample can be even monitored after each cutting, until salt alkali content is down to expected water in soil It is flat.
According to embodiment preferred for this invention, wherein the planting density of the rehabilitation plant is calculated as with seed amount 300-600/m2
According to embodiment preferred for this invention, it is described to soil apply repair biological agent be by biological agent to kind Son, which is coated, to be applied, and/or directly biological agent is applied in soil.
According to embodiment preferred for this invention, described apply to soil repairs biological agent including that will repair biology system Agent is applied by way of being coated to seed comprising before planting rehabilitation plant, is coated and is repaired to vegetable seeds Biological agent.
Wherein preferred microbial inoculum coating amount is by 5~10ml/kg seeds.
It wherein more preferably repairs biological agent coating amount and 1~20,000 cells/grain is calculated as with the plant growth-promoting bacteria wherein contained.
According to another preferred embodiment of the present invention, also it is containing mass percent concentration in the reparation biological agent 0.5% methylcellulose.
According to embodiment preferred for this invention, the seed pelleting can use existing commercially available seed pelleting machine.
The preparation of the microbial inoculum is referred to the preparation method of patent 200810181327.8, wherein being specially:
(1) takes 1 gram of plant rhizosphere soil, immerses in 50ml PAF extraction solution, then be placed in 250ml containers;The PAF Extraction solution is to contain proteose peptone, casein hydrolysate, MgSO4、K2HPO4With the composition solution of glycerine;
(2) shakes the above-mentioned triangular flask that solution is extracted containing 50ml PAF at 25 ± 2 DEG C;
(3) the PAF extraction solution after takes 1ml above-mentioned steps (2) to shake, then dissolve in and extract solution containing 50ml PAF In 250ml containers, at 25 ± 2 DEG C, concussion;
(4) takes the PAF extraction solution that 1ml above-mentioned steps (3) obtain, and dissolves in the 250ml containing 50ml DF salting liquids and holds In device, shaken at 25 ± 2 DEG C;The DF salting liquids are to contain dipotassium hydrogen phosphate, disodium hydrogen phosphate, magnesium sulfate, glucose, glucose The solution of acid, citric acid, ferrous sulfate, boric acid, manganese sulfate, zinc sulfate, copper sulphate, molybdenum trioxide and ammonium sulfate;
(5) takes the extraction solution that 1ml above-mentioned steps (4) obtain, then dissolves in 50ml without containing ammonium sulfate but contain 3.0mM In the container of the above-mentioned DF salting liquids of 1- amino-cyclopropane -1- carboxylic acids, shaken at 25 ± 2 DEG C;
(6) takes the extraction solution that 0.01ml above-mentioned steps (5) obtain, and accesses the tablet of the DF salt containing 0.03mM ACC On gel entrapment culture base, cultivated 2-4 days at 30 ± 2 DEG C;
(7) takes the bacterium colony grown on the culture mediums of step (6), is plant growth-promoting bacteria Pseudomonas cedrina The strain of UW3 and Pseudomonas putida UW4;
(8) takes the strain that above-mentioned steps 7 obtain, and is seeded in pancreas peptone soybean broth culture medium, in 25 ± 2 DEG C of temperature The lower culture concussion of degree 24 hours, obtains culture solution;
(9) culture solution of centrifugation steps (8) at 4 DEG C;
(10) cell of DF liquid rinsing step (9) centrifugations;The ingredient of the DF liquid include sucrose, succinic acid, EDTA, dipotassium hydrogen phosphate, potassium dihydrogen phosphate and magnesium chloride;
(11) the DF cell solutions that centrifugation step (10) obtains again at 4 DEG C;
(12) dissolves the cell of centrifugation with DF saline solutions, and the ACC of 3.0mM is added, obtains mycetocyte DF salting liquids;
(13) the mycetocyte DF salting liquids that steps (12) obtain are cultivated at 25 ± 2 DEG C, are shaken;
(14) the DF culture solutions that is obtained in 4 DEG C of centrifugation steps (13), obtain the cell of centrifugation;
(15) it is molten to obtain mycetocyte DF salt for the cell for the centrifugation that is obtained with 10ml DF saline solutions dissolving steps (13) Liquid;
(16) the mycetocyte DF salting liquids that obtains step (15) inoculation fermentation at 28 ± 2 DEG C, obtains industrial fermentation The bacterium solution of plant growth-promoting bacteria afterwards;Bacterium number reaches hundred million/ml of 20-30 in the bacterium solution of the plant growth-promoting bacteria.
According to embodiment preferred for this invention, wherein described is that will directly repair to soil application reparation biological agent Biological agent is applied in soil;
Wherein preferably it is described repair biological agent be applied for seed manure or mouth fertilizer or base fertilizer application, applied amount be 50~ 100 kgs/acre.
According to embodiment preferred for this invention, wherein the reparation biological agent is applied for 50-100 kgs/acre.
On the other hand, the present invention also provides the rehabilitation plants and reparation biological agent to combine answering in soil remediation With the rehabilitation plant is annual ryegrass, English ryegrass, tall grass and meadow precocity grass original grass standing grain One or more combinations in undergraduate course crop;The reparation biological agent includes at least the bacterium solution containing plant growth-promoting bacteria, institute It states bacterium solution and contains plant growth-promoting bacteria Pseudomonas cedrina UW3 and Pseudomonas putida UW4.
The wherein preferably described grassland grass family effect includes rye or triticale.
The wherein present invention preferably soil remediation is the soil corroded by petroleum hydrocarbon.
In conclusion the present invention provides a kind of biological prosthetic oil hydrocarbon compound contaminated soil method and applications.This The method of invention has the following advantages that:
(1) green environmental protection technique of solar energy is utilized;
(2) applicability is extensive, is applicable to different regions and Different climate condition;
(3) keep and restore the natural structure and physics of soil, chemistry and biology feature;
(4) of low cost, implement simply, it is highly practical;
(5) processing of a variety of pollution mixed compounds can be carried out at the same time;
(6) environmentally safe, secondary pollution and any environment hidden danger are not stayed;
(7) a large amount of bio-energies can be provided, quickly, completely, thoroughly complete repair process.
The present invention can so that plant growth is vigorous by plant growth-promoting bacteria, so as to cause naturally occurring microorganism it is big Amount proliferation, and further promote enlivening for root system of plant, largely to absorb the pollutants such as the inorganic salts in soil;
The method of the present invention can greatly shorten time of soil remediation, can be with usually in a seriously polluted area It controls and is repaired to the level before pollution in 3-5.
Description of the drawings
Fig. 1 is the repair process analysis diagram for formulating the method for repairing oil hydrocarbon compound contaminated soil;
Fig. 2 is the reparation result schematic diagram of embodiment 1;
Fig. 3 is the reparation result figure of embodiment 2;
Fig. 4 is the reparation result figure of embodiment 3;
Fig. 5 is the reparation result figure of embodiment 4.
Specific implementation mode
Below by way of the advantageous effect of the specific embodiment implementation process that the present invention will be described in detail and generation, it is intended to which help is read Reader more fully understand the present invention essence and feature, not as to this case can practical range restriction.
Embodiment 1
The repair process of the soil of central plain area (Henan Province) oil pollution, process substantially may refer to shown in Fig. 1.Soil Heavy crude containing 5600ppm;The typical farmland in locality that area is 1.5 hectares;Crops (wheat and corn) are not growing; 2 months 2010 site preparations, smooth, apply fertilizer (N:10 kgs/acre), it applies and repairs biological agent (100 kgs/acre) and rotary tillage;It is broadcast at the beginning of 3 months Kind, annual ryegrass (Lolium multiflorum), English ryegrass (Lolium perenne), tall grass (Festuca arundinacea) is with 2:1:1 ratio mixes, 16 kilograms of mu dosage;Seed carries out bacterium solution coating before broadcasting, coating Bacterium is Pseudomonas cedrina UW3 and Pseudomonas putida UW4 fermentation bacterium stoste (patents 200810181327.8 embodiments 1 prepare the bacterium solution obtained to step (16)), by 8ml/kg seed pelletings, 20,000 bacterium cells/grain; In 8 months repair processes, 2 times are extracted, 6 and by the end of August;(30) analytical control is sampled by the end of October, is as a result summarized as follows:New skill The degradation rate 91% of art, traditional microbiological technology are 31%.As shown in Figure 2.
Embodiment 2
The repair process of the Northeast (Liaoning Province) coal tar contaminated soil.Soil contains the coal tar greasy dirt of 3100ppm Dye;Area is 2.1 hectares;Crops (wheat and corn) cannot grow;In March, 2011, site preparation, smooth, and apply fertilizer (N:10 kilograms/ Mu), it applies and repairs biological agent (100 kgs/acre) and rotary tillage;Sowing, annual ryegrass (Lolium at the beginning of 4 months Multiflorum), English ryegrass (Lolium perenne), tall grass (Festuca arundinacea) are with 2:1: 1 ratio mixes, 20 kilograms of mu dosage;Seed carries out bacterium solution coating before broadcasting, and coating bacterium solution is Pseudomonas cedrina UW3 and Pseudomonas putida UW4,20,000 bacterium cells/grain;In 6 months repair processes, 2 times are extracted, 6 and by the end of August; (30) analytical control is sampled at the beginning of 10 months, is as a result summarized as follows:The degradation rate 82% of new technology, traditional microbiological technology are 43%, traditional non-microorganism technology is 35%;Especially to the degradation of polycyclic aromatic hydrocarbon, include the drop of the above aromatic hydrocarbons of 5 rings of acene arsenic Solution is also 50% or more, the above aromatic hydrocarbons of 5 rings and traditional technology can hardly degrade.Degradation rate is as shown in figure 3, wherein traditional non-life Object technology refers to the method ploughed using conventional mechanical, and traditional biological technology is to add conventional mechanical using existing microbial technique It ploughs method.Wherein each ingredient abbreviation indicates as follows:
16 kinds of common polycyclic aromatic hydrocarbons
Naphthalene naphthalenes (NAP)
Acenaphthylene acenaphthylenes (ANY)
Acenaphthene acenaphthenes acenaphthene (ANA)
Fluorene fluorenes (FLU)
Phenanthrene phenanthrene (PHE)
Anthracene anthracenes (ANT)
Fluoranthene fluoranthene (FLT)
Pyrene pyrenes (PYR)
Benzo (a) anthracene benzos (a) anthracene (BaA)
Chrysene (CHR)
Benzo (b) fluoranthene benzos (b) fluoranthene (BbF)
Benzo (k) fluoranthene benzos (k) fluoranthene (BkF)
Benzo (a) pyrene benzos (a) pyrene (BaP)
Indeno (1,2,3-cd) pyrene indenos (1,2,3-cd) pyrene (IPY)
Dibenzo (a, h) anthracene dibenzo (a, h) anthracene (DBA)
Benzo (g, h, i) perylene benzos (ghi) (perylene) (BGP)
Embodiment 3:
The repair process of the Northeast (Heilongjiang Province) heavy oil contaminated soil.Soil contains the heavy oil pollution of 1200ppm (the mainly part of 15-25C molecules);Area be 0.5 hectare in April, 2009 site preparation, it is smooth, apply fertilizer (N:10 kgs/acre), it applies Repair biological agent (50 kgs/acre) and rotary tillage;May 20 was sowed, annual rye, tall grass (Festuca Arundinacea) with 3:1 ratio mixes, 20 kilograms of mu dosage;Seed carries out bacterium solution coating before broadcasting, and coating bacterium solution is Pseudomonas cedrina UW3 and Pseudomonas putida UW4,20,000 bacterium cells/grain;In reparation in 5 months Journey extracts in 1 time, 7 months;(3) analytical control is sampled at the beginning of October 20, is as a result summarized as follows:The degradation rate 55% of new technology, Traditional microbiological technology is 15%.The results are shown in Figure 4.
Embodiment 4:
The repair process of North China (Shandong Province) heavy oil contaminated soil.Soil contains the heavy oil of 15% (15000ppm) Pollution (the mainly part of 15-25C molecules);Area be 4 hectares in April, 2004 site preparation, it is smooth, apply fertilizer (N:20 kgs/acre), It applies and repairs biological agent (100 kgs/acre) and rotary tillage;May 20 was sowed, annual ryegrass (Lolium Multiflorum), English ryegrass (Lolium perenne), tall grass (Festuca arundinacea) are with 2:1: 1 ratio mixes, 20 kilograms of mu dosage;Seed carries out bacterium solution coating before broadcasting, and coating bacterium solution is Pseudomonas cedrina UW3 and Pseudomonas putida UW4,20,000 bacterium cells/grain;In 5 months repair processes, extract in 1 time, 7 months;October (3) analytical control is sampled at the beginning of 20 days, continuous 3 years reparation results are summarized as follows:Annual 4% petroleum wastewater of degradation rate of new technology Dye, 12% oil pollution of solution rate in 3 years, oil content are down to 3% by 15%.The results are shown in Figure 5.

Claims (15)

1. a kind of biological prosthetic oil hydrocarbon compound contaminated soil method, which is characterized in that the method includes in the soil It plants rehabilitation plant and soil is applied and repair biological agent, the rehabilitation plant is annual ryegrass, perennial rye The combination of grass and tall grass;The reparation biological agent includes at least the bacterium solution containing plant growth-promoting bacteria, and the bacterium solution contains Plant growth-promoting bacteria Pseudomonas cedrina UW3 and Pseudomonas putida UW4, the method further includes planting Soil is irrigated, cut off and is applied fertilizer after planting rehabilitation plant, wherein in the 60-70 days first times of rehabilitation plant growth course It cuts off, and later primary per cutting off within 40-60 days, cuts off rear toothing 10-12cm every time.
2. according to the method described in claim 1, it is characterized in that, in bacterium solution, the bacterium number ratio of UW3 and UW4 are 1:2~2: 1。
3. according to the method described in claim 1, it is characterized in that, the reparation biological agent further includes composite carrier, The composite carrier include organic matter, humic acid, decomposed organic matter, complexing displacer, in embedded material arbitrary two kinds with On combination.
4. according to the method described in claim 3, it is characterized in that, the ratio of composite carrier and bacterium solution is plant growth-promoting bacteria At least 0.2~200,000,000/every gram composite carrier of bacterium number.
5. according to the method described in claim 3, it is characterized in that, by the weight of composite carrier be 100% on the basis of, institute The composite carrier stated includes the ingredient of following weight percent:15~35% organic matter, 12~32% humic acid, 12 ~32% decomposed organic matter, 5~22% complexing displacer and 0.01~1% embedded material.
6. according to the method described in Claims 1 to 5 any one, which is characterized in that the method further includes being repaired in plantation To the pre-treatment step of soil before plant.
7. according to the method described in claim 6, it is characterized in that, the pre-treatment step include prepartion of land, except stone, turn over Ground, rake the soil level, apply fertilizer and sow before preparation one or more of.
8. according to the method described in Claims 1 to 5 any one, which is characterized in that the planting density of the rehabilitation plant with Seed amount is calculated as 300-600/m2
9. according to the method described in Claims 1 to 5 any one, which is characterized in that described apply to soil repairs biology system Agent is to be coated biological agent to seed to apply, and/or directly biological agent is applied in soil.
10. according to the method described in claim 9, it is characterized in that, described apply to soil repairs biological agent including that will give birth to Object preparation, which is coated seed, to be applied comprising before planting rehabilitation plant, is coated to vegetable seeds and is repaired biology system Agent;By 5~10ml/kg seeds.
11. according to the method described in claim 10, it is characterized in that, repairing plant of the biological agent coating amount wherein to contain Promoting bacteria is calculated as ten thousand cells of 1-2/grain.
12. according to the method described in claim 10, it is characterized in that, also containing mass percent in the reparation biological agent A concentration of 0.5% methylcellulose.
13. according to the method described in Claims 1 to 5 any one, which is characterized in that described apply to soil repairs biology system Agent is that directly biological agent is applied in soil.
14. according to the method for claim 13, which is characterized in that biological agent is applied for seed manure or mouth fertilizer or base fertilizer Using applied amount is 50~100 kgs/acre.
15. rehabilitation plant and reparation biological agent combine the application in soil remediation, the rehabilitation plant is annual rye The combination of grass, English ryegrass and tall grass;The reparation biological agent includes at least the bacterium containing plant growth-promoting bacteria Liquid, the bacterium solution contain plant growth-promoting bacteria Pseudomonas cedrina UW3 and Pseudomonas putida UW4;It is described Soil is by oil hydrocarbon compound contaminated soil, and the method for the soil remediation is included in after plantation rehabilitation plant to soil It irrigated, cut off and is applied fertilizer, wherein the 60-70 days in rehabilitation plant growth course are cut off for the first time, and later per 40- It cuts off within 60 days once, cuts off rear toothing 10-12cm every time.
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