CN105855290A - Fungus repair method for severe chromium-contaminated soil - Google Patents
Fungus repair method for severe chromium-contaminated soil Download PDFInfo
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- CN105855290A CN105855290A CN201610219557.3A CN201610219557A CN105855290A CN 105855290 A CN105855290 A CN 105855290A CN 201610219557 A CN201610219557 A CN 201610219557A CN 105855290 A CN105855290 A CN 105855290A
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- penicillium oxalicum
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- 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
Abstract
The invention discloses a fungus repair method for severe chromium-contaminated soil. According to the method, spores of Penicillium oxalicum SL2 are taken as a repair bacterial strain source, severe chromium-contaminated soil is mixed with a potato culture medium according to proportion to prepare a slurry suitable for germination and growth of bacterial strain spores, and a certain amount of oxalic acid is added to accelerate the repair efficiency of chromium contamination of the soil. The slurry inoculated with spores of Penicillium oxalicum SL2 is cultured in an environment of 30 DEG C, and after 10 days, the concentration of Cr (VI) in a soil leaching liquor is reduced to 0.7mg/L from 231.8mg/L, the removal rate reaches 99.7%, and the concentration of bioavailable form Cr (VI) in the repaired soil is reduced to 44.3mg/kg, which is reduced by 98.1% in comparison with soil treated by using a common clear water leaching method. The method disclosed by the invention is simple in operation, is environment-friendly, and can provide technical support for microbial remediation of severe chromium-contaminated soil.
Description
Technical field
The present invention relates to the fungi restorative procedure of a kind of severe chromium-polluted soil, belong to technical field of microbe application.
Background technology
China is to produce the country that chromic salts is most, and extensive and except chromium process the not execution in time of production technology result in the generation in a large amount of pollution of chromium place.According to statistics, China more than 100 at place by pollution of chromium, influence area is more than 50000 square kilometres, the level of pollution of some points of contamination positions exceeds standard 100 times, serious threat China ecological safety and population health, therefore, the most effectively repairing of chromium-polluted soil is extremely the most urgent.
Chromium mainly exists with Cr (III) and (VI) two kind of stable form of Cr after entering soil.Wherein Cr (VI) has teratogenesis, carcinogenicity and mutagenicity, and soluble in water, and in natural environment, transfer ability is extremely strong.And the water-soluble low transfer ability of Cr (III) is weak, being easily combined formation complex precipitate with organic matter, and be the necessary micro-of glucose, lipid and amino acid metabolism, toxicity is relatively low.Therefore, the basic ideas of soil Cr pollution control are that highly toxic Cr (VI) is reduced to Cr (III), reduce its healthy and ecological hazard.
The execution of traditional technology of repairing chromium contaminated soil such as chemical precipitation method, physical isolation method, ion-exchange etc. generally requires the substantial amounts of chemical reagent of consumption, and expensive parts cost of equipment and operating cost seriously hinder its popularization and application on a large scale.Meanwhile, pollutant removal is thorough, be easily generated the defects such as secondary pollution is also its restraining factors.And microorganism remediation technology is owing to cost is relatively low and the feature such as environmental friendliness, is increasingly favored.Find first from the seventies in last centuryP.dechromaticansCan reduce since Cr (VI), researcher is isolated and purified a large amount of chromium also pathogenic microorganism, such as achromobacterAchromobacterSp.Ch-1, microbacteriumMicrobacteriumSp.MP30, thiobacillus ferrooxidansAcidithiobacillus thiooxidansDeng.But mostly rest on the laboratory research stage, be difficult to carry out actual application.Trace it to its cause, the mainly feature that soil often has high salinity, chromium content is high in pollution of chromium place, seriously inhibiting growth metabolism and the reducing power of microorganism, therefore, how utilizing microbial technique to repair severe chromium-polluted soil becomes a difficult problem for research application at present.
This seminar once disclosed penicillium oxalicum and the screening technique (CN104560738A) thereof of a kind of reduction of hexavalent chromium, and the application is further application on this basis and expands.
Summary of the invention
In order to overcome the deficiencies in the prior art, the invention provides the fungi restorative procedure of a kind of severe chromium-polluted soil.
The purpose of the present invention is achieved through the following technical solutions:
The fungi restorative procedure of a kind of severe chromium-polluted soil, comprises the following steps:
1) makePenicillium oxalicumSL2 spore suspension: willPenicillium oxalicumSL2 cultivates 7-15 days in PDA culture medium flat plate, with the spore on water washing plate surface, can obtain after concussion dispersionPenicillium oxalicumThe spore suspension of SL2；
2) chromium-polluted soil is gathered；
3) preparation potato culture medium, its formula is: take fresh potato, peeling, digs up eye, cleans, section, claims 200g, cuts fritter, add 1000ml water boil 30min, double gauze filters, and filtrate supplies moisture to 1000ml, adds 10g glucose in filtrate, cooling after dissolving；
4) according to the concentration of soil poisoning Cr (VI), in 1%-25%(w/v) ratio chromium-polluted soil mixed with potato culture medium make mud；
5) in the mud of step 4), oxalic acid, final concentration 50mmol/L are added;
6) initial pH value of mud is adjusted to 4-8；
7) by 103～106Individual/L adds what step 1) obtained in the mud of step 6)Penicillium oxalicumSL2 spore suspension, at 30 DEG C, aeration is cultivated；
8), after repairing cultivation 7-15d, contaminated soil remediation completes.
Described step 2) in chromium-polluted soil air-dry grind after cross 20 mesh sieves.
Beneficial effects of the present invention:
The invention provides the fungi restorative procedure of a kind of severe chromium-polluted soil.The method withPenicillium oxalicum Severe chromium-polluted soil, for repairing bacterial strain kind source, is mixed the mud being adapted to bacterial strain spore germination and growth, and adds the small molecular organic acids such as a certain amount of oxalic acid to accelerate the remediation efficiency that soil Cr is polluted by the spore of SL2 in proportion with potato culture medium.To be vaccinated withPenicillium oxalicum The mud of SL2 spore is cultivated in 30 DEG C of environment, after 10 days, in soil extraction, Cr (VI) concentration drops to 0.7mg/L from 231.8mg/L, clearance reaches 99.7%, in soil after reparation, biologically effective state Cr (VI) concentration drops to 44.3mg/kg, processes soil than conventional clear water extraction and decreases 98.1%.The method of the present invention is simple to operate, environmental friendliness, can be that the microorganism remediation of severe chromium-polluted soil provides technical support.
Accompanying drawing explanation
Fig. 1. bePenicillium oxalicumSL2 photo in heavily contaminated Soil slurry；
Fig. 2. bePenicillium oxalicumSL2 and the scanning electron microscope (SEM) photograph of heavily contaminated soil particle；
Fig. 3. bePenicillium oxalicumSL2 processes the variation diagram of Cr (VI) in soil extract；
Fig. 4. it is pH pairPenicillium oxalicumSL2 removal Cr (VI) affects figure；
Fig. 5. it is inoculum concentration pairPenicillium oxalicumSL2 removal Cr (VI) affects figure；
Fig. 6. it is Cr (VI) initial concentration pairPenicillium oxalicumSL2 removal Cr (VI) affects figure；
Fig. 7. it is salinity pairPenicillium oxalicumSL2 removal Cr (VI) affects figure.
Detailed description of the invention
Embodiment 1 :A kind of fungi restorative procedure of severe chromium-polluted soil
(1) makePenicillium oxalicum SL2 spore suspension.WillPenicillium oxalicumSL2 cultivates 10 days in PDA culture medium flat plate, with the spore on water washing plate surface, obtains after concussion dispersionPenicillium oxalicum The spore suspension of SL2.Penicillium oxalicumSL2 is that the screening of this seminar obtains, and belongs to prior art, is stored in Chinese Typical Representative culture culture presevation administrative center, and preserving number is CCTCC NO:M2014505, on October 22nd, 2014.
(2) severe chromium-polluted soil picks up from retired chemical plant, Hangzhou on-site, this soil belongs to basic soil (pH is 10.2), soil organic matter content relatively low (5.16g/kg), Cation Exchange Capacity in Soils is 61.97cmol/kg, and total chromium content in soil is 7069.6mg/kg, and sexavalence lattice are 4385.7mg/kg, in addition, also having the heavy metal contaminants such as Pb, Cd and Cu in soil, concentration is respectively 584.3mg/kg, 10.4mg/kg, 52.7mg/kg.Contaminated soil is air-dried and after grinding, crosses 20 mesh sieves, be sub-packed in the conical flask of 250ml by 5g/ part, 121 DEG C of sterilizing 60min, after cooling down 1 day completely, in 121 DEG C of sterilizing 60min, be repeated 3 times.
(3) configuration potato culture medium, its formula is: take fresh potato, peeling, digs up eye, cleans, section.Claiming 200g, cut fritter, add 1000ml water boil 30min, double gauze filters, and filtrate supplies moisture to 1000ml.10g glucose is added, in 115 DEG C of sterilizing 20min after dissolving in filtrate.
(4) packet transaction contaminated soil as follows: A. water+sterile soil；B. potato culture medium+sterile soil；C.50mmol/L oxalic acid+sterile soil；D. 50mmol/L oxalic acid+potato culture medium+sterile soil；E. spore suspension+potato culture medium+sterile soil；F. 50mmol/L oxalic acid+spore suspension+potato culture medium+sterile soil.5%(w/v is obtained by above-mentioned process) mud, regulate pH to about 6, put into magnetite, thereinPenicillium oxalicumSL2 spore suspension presses 106Individual/L adds.Each process 5 repetition, cultivates in the environment of 30 DEG C.
(5) in incubationPenicillium oxalicumSL2 spore germination forms mycelium bead, such as Fig. 1.By the mycelial microstructure of scanning electron microscopic observation, as in figure 2 it is shown, mycelia is distributed in the surface of big soil particle, and some little soil particles adhere on mycelium.
(6) during cultivating, the concentration of Cr (VI) in leaching liquor is detected, result shows as shown in Figure 3, the process group effect that microorganism adds oxalic acid is best, wherein Cr (VI) concentration started to significantly reduce from second day, after 10 days, the Cr in soil extraction (VI) concentration drops to 0.7mg/L from 231.8mg/L, and clearance reaches 99.7%.
Dry repairing the soil after cultivating 12d to constant weight, the method extracted with DTPA detects the situation of change of wherein biologically effective state Cr (VI), result shows that in water extract method process group, contaminated soil biologically effective state Cr (VI) concentration is 2315.3mg/kg, microorganism adds biologically effective state Cr (VI) concentration in the process group soil of oxalic acid and is then reduced to 44.3mg/kg, reduces 98.1% than conventional water extract method.
Embodiment 2 :PH pairPenicillium oxalicumSL2 removes the impact of Cr (VI)
Cr (VI) solution of preparation 5g/L: weigh 7.5g potassium bichromate 110 DEG C and dry 2h, then weigh 7.073g and be dissolved in Cr (VI) solution making 5g/L in 500mL water.Cr (VI) solution of 5g/L is filtered, for regulating Cr (VI) concentration in culture medium with the aseptic filter of 0.22um.
Preparation potato culture medium: take fresh potato, peeling, dig up eye, clean, section.Claiming 200g, cut fritter, add 1000ml water boil 30min, double gauze filters, and filtrate supplies moisture to 1000ml.In filtrate, add 10g glucose, cool down after dissolving, be sub-packed in the triangular flask of 250ml by 50ml/, 115 DEG C of sterilizing 20min.
By Cr (VI) concentration in aseptic Cr (VI) solution regulation culture medium, the Cr (VI) in system is made to be about 160mg/L, pH to 3,4,5,6,7,8,9,11 are adjusted, in the change of different time detecting wherein Cr (VI) concentration with concentrated hydrochloric acid.Result as shown in Figure 4, when initial pH is 4-8,Penicillium oxalicumSL2 can effectively remove Cr (VI)；As pH=6,Penicillium oxalicumThe speed that SL2 removes Cr (VI) is relatively fast, is 88.7% at the 6th day Cr (VI) clearance cultivated, within the 7th day, reaches 96.0%.Result showsPenicillium oxalicumSL2 can process Cr (VI) in the environment of pH4～8 and pollute.
Embodiment 3 :Inoculum concentration pairPenicillium oxalicumSL2 removes the impact of Cr (VI)
Compound concentration is 108The spore suspension of individual/L, then to obtain concentration by the method for gradient dilution be 107Individual/L, 106Individual/L, 105The spore suspension of individual/L.Aseptic Cr (VI) solution is added so that in system, Cr (VI) initial concentration is about 160mg L in the triangular flask filling potato culture medium-1.By 1:100(V/V) inoculate the spore suspension prepared, it is processed as comparison, at 30 DEG C, shaken cultivation under the conditions of 200 rpm, Cr (VI) concentration in different time sampling and measuring nutrient solution with sterilized water.Result is as shown in Figure 5: at the 3rd day cultivated, the spore concentration of inoculation was the highest, and in system, the concentration of Cr (VI) is the lowest, but the Cr (VI) in the 5th day all process groups can be reduced.ShowPenicillium oxalicumSL2 spore inoculating amount is 103～106Individual/L time, can effectively process Cr (VI) pollute.
Embodiment 4 :Cr (VI) initial concentration pairPenicillium oxalicumSL2 removes the impact of Cr (VI)
Respectively toward the 250ml triangular flask fill 50ml potato culture medium adds 0ml, 0.625ml, 1.25ml, 3.75ml, 7.5ml, 12.5ml through filtration sterilization Cr (VI) mother liquor and 12.5ml, 11.875ml, 11.25ml, 8.75ml, 5ml, 0ml sterilized water, make Cr in cultivation (VI) final concentration of 0,50,100,300,600,1000mg/L.By 105Individual/L inoculationPenicillium oxalicumSL2 spore, in 30 DEG C, shaken cultivation under the conditions of 200 rpm, Cr (VI) concentration in different time sampling and measuring nutrient solution.Reaction is terminated after 9 days.Result is as shown in Figure 6, in the system of below 300mg/L, Cr (VI) can be reduced more than 99%, and Cr (VI) percent reduction is 61.7% in the system of 600mg/L, in the system for handling of 1000mg/L, Cr (VI) percent reduction is 40.4%, showsPenicillium oxalicumSL2 removes Cr (VI) has certain concentration range to limit, and needs to regulate the proportioning of mud when repairing Cr (VI) contaminated soil with it.
Embodiment 5 :Salinity pairPenicillium oxalicumSL2 removes the impact of Cr (VI)
Different salinity culture medium is prepared: adding 160ml in the beaker of 400ml and mix up the culture medium of pH, be separately added into 0,0.16,0.32,0.8,1.6,3.2,8,16 g NaCl, making containing NaCl concentration is 0,1,2,5,10,20,50,100 g L-1Potato culture medium.Being divided in the triangular flask of 250ml by culture medium, each 50ml, ventilated membrane seals, 115 DEG C of sterilizing 20min.
Aseptic Cr (VI) mother liquor is added so that in system, Cr (VI) initial concentration is about 200mg L in the potato culture medium prepared-1, by 106Individual/L inoculating spores, at 30 DEG C, shaken cultivation under the conditions of 200 rpm, Cr (VI) concentration in different time detection architecture.Result as it is shown in fig. 7, in all process groups Cr (VI) can be reduced, showPenicillium oxalicumSL2 can apply to the Cr (VI) of high salt and pollutes environment.
Claims (2)
1. the fungi restorative procedure of a severe chromium-polluted soil, it is characterised in that comprise the following steps:
1) makePenicillium oxalicumSL2 spore suspension: willPenicillium oxalicumSL2 cultivates 7-15 days in PDA culture medium flat plate, with the spore on water washing plate surface, can obtain after concussion dispersionPenicillium oxalicumThe spore suspension of SL2；
2) chromium-polluted soil is gathered；
3) preparation potato culture medium, its formula is: take fresh potato, peeling, digs up eye, cleans, section, claims 200g, cuts fritter, add 1000ml water boil 30min, double gauze filters, and filtrate supplies moisture to 1000ml, adds 10g glucose in filtrate, cooling after dissolving；
4) according to the concentration of soil poisoning Cr (VI), in 1%-25%(w/v) ratio chromium-polluted soil mixed with potato culture medium make mud；
5) in the mud of step 4), oxalic acid, final concentration 50mmol/L are added;
6) initial pH value of mud is adjusted to 4-8；
7) by 103～106Individual/L adds what step 1) obtained in the mud of step 6)Penicillium oxalicumSL2 spore suspension, at 30 DEG C, aeration is cultivated；
8), after repairing cultivation 7-15d, contaminated soil remediation completes.
Fungi restorative procedure the most according to claim 1, it is characterised in that described step 2) in chromium-polluted soil air-dry grind after cross 20 mesh sieves.
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| CN108441449A (en) * | 2018-04-04 | 2018-08-24 | 中南民族大学 | A kind of bacterial strain Y2 independently tamed and its application |
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Cited By (1)
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