CN101992207A - Heavy metal-polycyclic aromatic hydrocarbon combined pollution reinforcing repairing agent and application thereof - Google Patents
Heavy metal-polycyclic aromatic hydrocarbon combined pollution reinforcing repairing agent and application thereof Download PDFInfo
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Abstract
The invention relates to phytoremediation of environment pollution, in particular to a reinforcing repairing agent of a hyperaccumulator nightshade, which comprises the following components of glycine, glutamic acid and cysteine that have a mol ratio of 0.1-1.0:0.1-1.0:0.1-1.0. In cadmium-polycyclic aromatic hydrocarbon combined pollution pot culture soil in which the hyperaccumulator nightshade is transplanted, the glycine, the glutamic acid and the cysteine are mixed and poured in the soil in a form of a water mixture according to the content of 0.1-1.0mmol/kg every 15 days for 3 times; and when the hyperaccumulator nightshade grows up, the hyperaccumulator nightshade and the soil sample are harvested. Compared with the reinforcing repairing agent without being added with amino acid, low quantity of the mixture of the glycine, the glutamic acid and the cysteine has the best treatment effect, the Cd content of the upper part of the nightshade soil is increased by 2.26 times, and the removal rate of the total amount of polycyclic aromatic hydrocarbon in the soil reaches 34.5 percent, thus the repairing efficiency of the Cd and the polycyclic aromatic hydrocarbon by the nightshade are remarkably improved.
Description
Technical field
The present invention relates to the phytoremediation technology of heavy metal-polycyclic aromatic hydrocarbon combined contamination soil, specifically a kind of reinforcement renovation agent of super enrichment repair plant black nightshade.
Background technology
Variation along with pollutant emission, complicated and frequentization, the contaminated degree of major receptors soil of pollutant increases the weight of day by day, also outstanding all the more (the document 1:Paton GI of the rational feature of complexity that contaminated soil is repaired and refractory, Viventsova E, Kumpene J, et al.2006.An ecotoxicity assessment of contaminated forest soils from the Kola Peninsula.Science of the Total Environment, 355 (13): 106-117), wherein heavy metal-organic combined pollution is more common pollution type, as Cd-polycyclic aromatic hydrocarbon (Polycyclic aromatic hydrocarbons, PAHs) combined pollution.Cadmium when low concentration can by reduce photosynthesis of plant, produce approach such as oxidative stress suppress the plants growth, to plant produce toxic action (document 2:Schill RO,
H.
HR.2003.Laboratory simulation of a mining accident:acute toxicity hsc/hsp70 response and recovery from sress in Gammarus fossarum (Crustacea smphipoda) exposed to a pulse of cadmium.Biometals, 16:391-401), and to some extent humans and animals is produced toxic action, also can bring out canceration; And PAHs is a class persistence organic pollutant, and biology is had chronic toxicity and carcinogenic, teratogenesis, mutagenicity.Because Cd, PAHs show as in soil environment and have disguise and chronicity, cause the difficulty of repairing this type of combined contamination soil to strengthen, seeking efficient, safe, economic recovery technique becomes the realistic problem of needing solution badly.Phytoremediation technology have physics, chemical repair method incomparable expense cheap, to be difficult for causing secondary pollution, to beautify the environment, be easy to be advantages such as society is accepted, become recovery technique (the document 3:Chaney RL that has development potentiality, Malik M, Li YM.1997.Phytoremediation of soil metals.Current Opinions in Biotechnology, 8:279-284; Brooks RR, Chambers MF, Nickls L J, et al.1998.Phytomining.Trends in Plant Science, 3 (9): 359-362).The phytoremediation of heavy-metal contaminated soil mainly is to remove by the excess of super enriching plant or enriching plant to be used for realizing (document 4: Wei Shuhe, Zhou Qixing, Pavel V.Koval, Galina A.Belogolova. organic contamination environment phytoremediation technology. ecological magazine .2006,25 (6): 716-721); For organic contamination such as polycyclic aromatic hydrocarbon mainly is to utilize the degradation of plant rhizosphere circle microorganism to finish (document 4: Wei Shuhe, Zhou Qixing, Pavel V.Koval, Galina A.Belogolova. organic contamination environment phytoremediation technology. ecological magazine .2006,25 (6): 716-721).But it is restricted with some that phytoremediation technology also exists some defectives, and the own biological characteristics of environmental condition and plant influences as very easily being subjected to, repairing efficiency is long etc.So need to adopt some to strengthen the efficient that reclamation activities improves phytoremediation, in the hope of providing reference for improving early of this technology.
Coerce down in heavy metal pollution; a lot of plants have formed the harm that certain self-protective mechanism is resisted heavy metal ion; as a series of little molecule (citrates of biosynthesis with complexation heavy metal ion ability; glutathione; plant complexing element etc.); form the lower compound of toxicity with free heavy metal ion complexing, and it is stored in the vacuole of plant cell.Wherein plant complexing element is an important detoxifcation material in the plant. plant complexing element is a class sulfydryl chelating polypeptide that produces in the cytoplasm liquid under the stress-inducings such as heavy metal such as Ni, Cd, Zn, Te, Au, Hg, Pb, As, Se, wherein Cd is the strongest interior inducible factor of body, and the performance inducing action be metal ion free in the cytoplasm (document 5:Rauser WE.1995.Phytochelatins and related peptides.Plant Physiology, 109:1141-1149).PCs (Phytochelatins) is the polypeptide that a class is rich in cysteine (cysteine), is that original substrate generates through a series of enzymatic reactions by cysteine, glutamic acid, glycine.Cys sulfydryl among the PCs can with the cd in the born of the same parents
2+Form coordinate bond, and then formation PCs-Cd compound, the toxicity of this compound pair cell greatly reduces, and plant has also further taked vacuole compartmentation (compartmentation) that compound is fixed in the vacuole in addition, has further reduced the toxic action of Cd to plant.But under Cd-PAHs combined pollution condition, the original substrate of external source PCs absorbs black nightshade and the feature of accumulation Cd and to the influence of degrading polycyclic aromatic hydrocarbons effect in the soil, yet there are no report, and this also is the problem that phytoremediation technology presses for solution.
Summary of the invention
The object of the present invention is to provide a kind of result of use heavy metal-polycyclic aromatic hydrocarbon combined contamination soil good, that can improve the crop remediation efficiency to strengthen renovation agent and application thereof.
For achieving the above object, the technical solution used in the present invention is:
One heavy metal species-polycyclic aromatic hydrocarbon combined contamination soil is strengthened renovation agent, and its composition is calculated in molar ratio as, glycine: glutamic acid: cysteine=0.1-1.0: 0.1-1.0: 0.1-1.0.
Described reinforcement renovation agent can add in kind and be implanted with in the heavy metal-polycyclic aromatic hydrocarbon combined contamination soil of super enriching plant and/or enriching plant, to improve super enriching plant (black nightshade) and/or the enriching plant remediation efficiency to heavy metal such as cadmium and polycyclic aromatic hydrocarbon.
Described reinforcement renovation agent glycine, glutamic acid or the cysteine concentration that adds in contaminated soil is respectively 0.1-1.0mmol/kg soil, preferably is respectively 0.2-0.6mmol/kg soil.
On the soil of land for growing field crops or Polluted area, when directly applying the present invention and strengthening renovation agent, strengthen the every kg soil that adds in concentration adopted of renovation agent in contaminated soil, it is with the soil weight metering of the 0-20 centimetre of dark soil thickness in soil layer surface.
. the content 0.3-100mg/kg of heavy metal Cd in the described contaminated soil, the content 0.3-600mg/kg of polycyclic aromatic hydrocarbon PAHs; Described super enriching plant is a black nightshade.
The present invention mixes with the form of water mixed liquid by the content of 0.1-1.0mmol/kg on the contaminated soil of plantation black nightshade and pours into glycine, glutamic acid and cysteine, when treating that black nightshade is ripe, results plant and soil-like, with do not add amino acid whose contrast and compare, the glycine of low dosage, glutamic acid and cysteine mixed liquor treatment effect the best, black nightshade overground part Cd content has increased by 2.26 times, simultaneously, the clearance of soil polycyclic aromatic hydrocarbon total amount can reach 39.3%, has improved the remediation efficiency of black nightshade to cadmium and polycyclic aromatic hydrocarbon significantly.
Description of drawings
Fig. 1 exogenous amino acid is to the influence of black nightshade plant height;
Fig. 2 exogenous amino acid is to the influence of black nightshade overground part dry weight.
The specific embodiment
The embodiment exogenous amino acid is to the reinforcement repair of heavy metal-polycyclic aromatic hydrocarbon combined pollution vegetable field soil
1 test material
The combined pollution vegetable field soil picks up from the Shen of Shenyang City's Dongling District and comforts the irrigated area, the vegetables field of irrigating for many years for petroleum waste water (be the paddy field in the past, and changed the vegetables field into before 10 years).Soil types is a yellowish soil, and its physicochemical property is a pH value 6.9, organic 17.95mgkg
-1, full N 1.32mgkg
-1, full P 1.39mgkg
-1, quick-acting K 144.36mgkg
-1The total cadmium content 0.711mgkg of soil
-1, 2 grade standards above national soil environment quality GB15618-1995 belong to the cadmium pollution horticultural vegetable field soil; 16 kinds of polycyclic aromatic hydrocarbon total amounts are 0.569mgkg in the soil
-1, 2-4 ring polycyclic aromatic hydrocarbon content is 0.462mgkg
-1, 5-6 ring polycyclic aromatic hydrocarbon content is 0.106mgkg
-1, benzo (a) pyrene content is 0.026mgkg
-1, compare with Dutch soil remediation desired value, gather that polycyclic aromatic hydrocarbon content obviously exceeds standard in the soil.
Rehabilitation plant is black nightshade (Solanum nigrum), and seed picks up from the non-dirty agricultural soil of irritating in ecological experiment station, Chinese Academy of Sciences Shenyang.
2 test methods
According to synthetic level of PCs in the plant and the effect in the detoxification processes in plant thereof, experimental design cysteine (Cys), glycine (Gly), the single processing of glutamic acid (Glu), reach glycine (Gly)+glutamic acid (Glu), glycine (Gly)+glutamic acid (Glu)+cysteine (Cys) Combined Processing, form with water mixed liquid pours into, adding concentration range is 0.1-1.0mmol/kg, be divided into two levels of low dosage 0.3mmol/kg and high dose 0.9mmol/kg, specifically see Table 1.Other establishes 1 and does not add amino acid whose contrast, totally 11 processing, and every processing repeats 3 times (pouring into altogether 3 times), adopts liquid to add method, divides to add for three times, and interval of adding is 15 days.
Test is carried out in the greenhouse.To cross the 4mm sieve for the air-dry back of examination soil, accurately the weighing 1kg soil plastic tub of packing into; Black nightshade seed is seeded in Miao Panzhong, when treating that the black nightshade seedling grows to 3-5 sheet spire, select the consistent black nightshade seedling of growth to move into respectively and respectively handle in the basin, every potted plant 2 seedling. control the temperature 25-30 of culturing room ℃, air humidity 50%-80%, intensity of illumination 350-450umol/m
2S, light application time 08:00-17:30.Transplant after 15 days, when treating that seedling grows to the 10cm left and right sides, corresponding each processing adds respectively, and irregularly waters running water according to basin lack of water situation, makes soil moisture content often remain on about 80% of field capacity.For preventing the filter loss of treatment fluid leaching, under basin, place plastic pallet and also in time effusion is refunded in the basin.After 80 days in plant maturity sampling (when treating that promptly black nightshade is ripe, results plant and soil-like).
Table 1 cysteine, glycine, glutamic acid processing horizontal table (mmol/kg)
3 sample analysis
The plant sample part of results is used for the mensuration of overground part content of beary metal, and remainder root division, stem, leaf, fruit 4 parts are fully washed with running water respectively, and then use deionized water rinsing, the drop branch that anhydrates, the 30min that completes under 105 ℃ dries to constant weight in baking oven under 70 ℃ then.Plant sample crushing screening after the oven dry is standby, and it is standby that 100 mesh sieves are crossed in the air-dry back of pedotheque.Plant reaches upward, and the earth sample all adopts HNO
3-HClO
4Method digestion (the two volume ratio is 3: 1), the atomic absorption spectrophotometer method is measured the content of Cd, and atomic absorption spectrophotometer is the 180-80 of Hitachi, and the absorbing wavelength of cadmium is 228.8.The mensuration of basic physical and chemical such as the content of organic matter of soil adopts conventional assay method.
100 mesh sieves are crossed in the air-dry back of the pedotheque of results, are stored in-80 ℃ of refrigerators standby.P in soil AHs extracting method is with reference to Song Yufang (1995) and Gao Yanzheng (2005).Take by weighing the 5.0g pedotheque in 25ml glass centrifuge tube,, adopt the mode of changing water to control therebetween and extract temperature below 40 ℃, with 4000rmin with the carrene ultrasonic extraction 2h of 15ml
-1Centrifugation is also drawn supernatant evaporate near doing in Rotary Evaporators, adds the cyclohexane dissolving of 1.00ml again, accurately draws 0.5ml and crosses the 1g silicagel column and purify, with n-hexane-carrene (volume ratio is 1: 1) mixed liquor wash-out of 11ml.Begin behind the 1ml eluent before discarding to collect, with the eluent collected in 40 ℃ of constant temperature down rotation be evaporated to driedly, again with the acetonitrile dissolving and be settled to 1ml, cross and go up machine behind the 0.45 μ m organic facies filter membrane and measure.
The mensuration of PAHs adopts fluorescence-ultraviolet series connection detection-high performance liquid chromatography (Rao Zhu etc., 2007).HPLC is Waters 1525, fluorescence detector (Multi λ Fluorescence Detector) is Waters 2475, UV-detector (Dual λ Absorbance Detector) is Waters2487, chromatographic column be the polycyclic aromatic hydrocarbon chromatographic column specially (4.6 * 250mm, 5um).Analysis condition: with the acetonitrile/water is that (0-3min acetonitrile/water volume ratio is 60: 40 to eluent gradient drip washing 36min; 3-30min acetonitrile/water volume ratio is 100: 0; 30-36min acetonitrile/water volume ratio is 60: 40).Flow velocity 1.00mlmin
-1, sample size is 20ul, 25 ℃ of column temperatures, and the corresponding mensuration wavelength of 16 kinds of PAHs is referring to (2007) such as Rao Zhu.With 16PAHs mixing standard specimen is external standard, directly contrasts with retention time and peak area and carries out the qualitative and quantitative of PAHs.This method is linear better to the mensuration of 16 kinds of PAHs, and the correlation coefficient r value is all greater than 0.999, and the scope of detecting is 0.011-0.513ng, to the recovery of standard addition of 16 kinds of PAHs except that the naphthalenes of 2 rings be 53.4% all the other all between 75%-110%.
4 data statistics
The data that obtain are carried out statistical analysis with Microsoft Excel and SPSS16.0, utilize the new multipole difference check of Duncan to carry out significance test of difference (p＜0.05).
5 results
5.1 exogenous amino acid is to the influence of black nightshade growth characteristics
Fig. 1, Fig. 2 have provided black nightshade plant height and the upperground part biomass situation of change under the indoor pot condition respectively.From in appearance, under Cd-PAHs combined pollution condition, minus green, the symptom of being coerced such as withered and yellow do not appear in black nightshade normal growth, blade.Obviously, heavy metal and polycyclic aromatic hydrocarbon do not produce inhibitory action to the normal growth of plant under this experiment condition.Plant plant height and overground part dry weight show as corresponding to rule to the reaction of additive.Do not compare with adding amino acid whose contrast, single factor of cysteine (Cys), glycine (Gly), each concentration level of glutamic acid (Glu) is handled plant plant height and overground part dry weight is had no significant effect (p＜0.05); And each Combined Processing all has significant facilitation (p＜0.05) to it.
5.2 exogenous amino acid is to absorbing the influence of Cd in the black nightshade body
Table 2 has provided different aminoacids and has handled black nightshade root, stem, leaf, fruit and 5 portion C d of overground part content down.As can be seen from Table 2, each is handled, and Cd content conspicuousness increases outer (p＜0.05) in the disleaf, and the content increase of root, stem, fruit and overground part all reaches utmost point significance (p＜0.01).The cumulant of plant shoot Cd still shows stronger extraction and accumulation ability greater than its cumulant at underground part under low concentration Cd pollutional condition, and different aminoacids handles and can significantly strengthen black nightshade Cd is extracted and accumulation ability, as handling 0.3mmolkg
-1Gly+0.3mmolkg
-1Glu+0.3mmolkg
-1Cys makes the comparison of black nightshade overground part Cd content according to having increased by 2.26 times.On the whole, the facilitation that the complex factors of variable concentrations level are handled the plant absorbing heavy metal is better than single factor processing, and 3 factor treatment effects are better than 2 factors to be handled, illustrate when adding the synthetic required whole amino acid of PCs and can improve its growing amount in plant, promoted the absorption of plant Cd.
Table 2 exogenous amino acid is to the influence of Cd accumulation in the black nightshade body
Super enriching plant (Hyperaccumulator) is meant accumulation of energy excess or the plant that accumulates several heavy metal elements simultaneously, and possesses critical value content, transfer and enrichment characteristics, and its transfer and enrichment characteristics mainly characterize by transfer ratio and concentration coefficient.Transfer ratio (translocation factor, TF) be meant the content of overground part element and the ratio of underground part constituent content of the same race, being used for estimating the ability of plant transhipment heavy metal. transfer ratio is big more, and then heavy metal is strong more to overground part organ turn-over capacity from root.For super enriching plant, its overground part content of beary metal is greater than the root content of beary metal.As can be seen from Table 3, under low concentration Cd-PAHs combined pollution condition, each handles TF＞1, and black nightshade still possesses stronger transfer ability to low concentration Cd.Concentration coefficient (bioaccumulation factor, BF) also claim absorption coefficient, be meant the ratio of certain constituent content and elements in Soil content in the plant, characterize the complexity of migration of element in the soil-plant system, concentration coefficient is high more, shows that plant shoot heavy metal accumulation mass fraction is big more.In soil, add the PCs substrate and can strengthen the absorption of plant shoot, when adding low dosage 3 seed amino acid intermixtures, make the ability of plant absorbing heavy metal improve 2.26 times heavy metal.The size of phytoremediation heavy metal ability and content and the upperground part biomass of plant heavy metal of body have direct relation, and (metal extraction ratio MER) characterizes to use the heavy metal recovery rate usually.The heavy metal recovery rate is meant in the amount of plant shoot accumulation heavy metal and the soil ratio with the heavy metal species amount, promptly
MER=(C
The overground part heavy metal* M
The overground part dry weight/ C
Heavy metal in soil* M
Soil weight) * 100
As shown in Table 3, adding of the PCs substrate of variable concentrations level improved the recovery rate of black nightshade to low concentration cadmium in the soil to a great extent, wherein cysteine is handled and is made the heavy metal recovery rate reach 1.80% and 1.83% respectively, has improved 1.80 and 1.83 times compared with the control respectively.In a word, under low concentration Cd-PAHs combined pollution condition, higher transfer ratio, concentration coefficient and heavy metal recovery rate are arranged all in the black nightshade body, demonstrate to Cd extract preferably, transfer and enrichment characteristics; Adding of PCs substrate can be strengthened the absorbability of black nightshade to Cd in the soil, quickens the removal speed of Cd in soil environment.
Table 3 exogenous amino acid is to the influence of concentration coefficient, transfer ratio and the heavy metal recovery rate of Cd
5.3 exogenous amino acid is to the influence of PAHs degraded in the soil
Table 4 has provided the influence of amino acid to degrading polycyclic aromatic hydrocarbons effect in the soil.As can be seen from Table 4, exogenous amino acid will be higher than the low-molecular-weight polycyclic aromatic hydrocarbon to the facilitation of polycyclic aromatic hydrocarbon with high molecular weight degraded in the soil, its reason may be that exogenous amino acid provides common metabolism matrix for edaphon, and metabolism is the biodegradable main mode of polycyclic aromatic hydrocarbon with high molecular weight altogether; In addition, the corresponding enhancing of degradation along with the increase 5-6 ring polycyclic aromatic hydrocarbon and benzo (a) pyrene of glycine dosage illustrates that glycine can provide the required C of common metabolism the source for the degraded of polycyclic aromatic hydrocarbon with high molecular weight.
Table 4 exogenous amino acid is to the enhancing degradation effect of PAHs in the soil
aNaphthalene, acenaphthene, acenaphthylene, fluorenes, anthracene, phenanthrene, fluoranthene, pyrene, benzo (a) anthracene,
Benzo (b) fluoranthene, benzo (k) fluoranthene
bBenzo (a) pyrene, and dibenzo (a, h) anthracene, benzo (g, h, i) perylene, indeno (1,2,3-cd) pyrene
Relatively can draw by table 2 and table 4, the facilitation that multifactor processing is repaired the Cd-PAHs contaminated soil to black nightshade is better than single factor processing, and handle T9, it is the low dosage glycine, glutamic acid and cysteine (Gly+Glu+Cys) mixed liquor is to removal effect the best of above two kinds of polluters, make black nightshade overground part cadmium content reach 5.65mg/kg, concentration coefficient is up to 7.9, and the cadmium recovery rate improves 2.26 times compared with the control; At this moment, the clearance of polycyclic aromatic hydrocarbon also reaches and is up to 34.5% in the soil.As seen the Gly+Glu+Cys of low dosage handles and has not only satisfied the synthetic required substrate requirement of PCs of plant, can also provide required matrix substrate for the soil microbial degradation polycyclic aromatic hydrocarbon.
Claims (7)
1. one heavy metal species-polycyclic aromatic hydrocarbon combined contamination soil is strengthened renovation agent, and it is characterized in that: its composition is calculated in molar ratio as, glycine: glutamic acid: cysteine=0.1-1.0: 0.1-1.0: 0.1-1.0.
2. the application of the described reinforcement renovation agent of claim 1 is characterized in that: described reinforcement renovation agent adds in kind and is implanted with in the heavy metal-polycyclic aromatic hydrocarbon combined contamination soil of super enriching plant and/or enriching plant.
3. according to the application of the described reinforcement renovation agent of claim 2, it is characterized in that: described reinforcement renovation agent glycine, glutamic acid or the cysteine concentration that adds in contaminated soil is respectively 0.1-1.0mmol/kg soil.
4. according to the application of the described reinforcement renovation agent of claim 2, it is characterized in that: described reinforcement renovation agent glycine, glutamic acid or the cysteine concentration that adds in contaminated soil is respectively 0.2-0.6mmol/kg soil.
5. according to the application of the described reinforcement renovation agent of claim 4, it is characterized in that: described reinforcement renovation agent adds the every kg soil of concentration in contaminated soil, and it is with the soil weight metering of the 0-20 centimetre of dark soil thickness in soil layer surface.
6. according to the application of the described reinforcement renovation agent of claim 2, it is characterized in that: the content 0.3-100mg/kg of heavy metal Cd in the described contaminated soil, the content 0.3-600mg/kg of polycyclic aromatic hydrocarbon PAHs.
7. according to the application of the described reinforcement renovation agent of claim 2, it is characterized in that: described super enriching plant is a black nightshade.
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