CN110018300A - The compound prediction technique and its application to wheat root elongation toxicity in soil of cadmium nickel - Google Patents
The compound prediction technique and its application to wheat root elongation toxicity in soil of cadmium nickel Download PDFInfo
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- 231100000419 toxicity Toxicity 0.000 title claims abstract description 64
- 230000001988 toxicity Effects 0.000 title claims abstract description 64
- 238000000034 method Methods 0.000 title claims abstract description 47
- OJIJEKBXJYRIBZ-UHFFFAOYSA-N cadmium nickel Chemical compound [Ni].[Cd] OJIJEKBXJYRIBZ-UHFFFAOYSA-N 0.000 title claims abstract description 31
- 150000001875 compounds Chemical class 0.000 title claims abstract description 25
- 230000000694 effects Effects 0.000 claims abstract description 79
- PXHVJJICTQNCMI-UHFFFAOYSA-N nickel Substances [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 claims abstract description 77
- 150000001768 cations Chemical class 0.000 claims abstract description 67
- 239000002131 composite material Substances 0.000 claims abstract description 48
- 238000002474 experimental method Methods 0.000 claims abstract description 39
- 229910052793 cadmium Inorganic materials 0.000 claims abstract description 37
- 230000003204 osmotic effect Effects 0.000 claims abstract description 37
- 229910052759 nickel Inorganic materials 0.000 claims abstract description 36
- 239000004576 sand Substances 0.000 claims abstract description 27
- 231100000331 toxic Toxicity 0.000 claims abstract description 23
- 229910001385 heavy metal Inorganic materials 0.000 claims abstract description 22
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- 230000007096 poisonous effect Effects 0.000 claims abstract description 14
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- 231100000783 metal toxicity Toxicity 0.000 claims abstract description 4
- 239000003446 ligand Substances 0.000 claims description 33
- 238000012545 processing Methods 0.000 claims description 25
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- 231100000820 toxicity test Toxicity 0.000 claims description 20
- 238000004364 calculation method Methods 0.000 claims description 17
- 230000000007 visual effect Effects 0.000 claims description 11
- 231100000916 relative toxicity Toxicity 0.000 claims description 8
- 238000009395 breeding Methods 0.000 claims description 7
- 230000001488 breeding effect Effects 0.000 claims description 7
- 238000012360 testing method Methods 0.000 claims description 7
- 230000008859 change Effects 0.000 claims description 6
- 238000005259 measurement Methods 0.000 claims description 6
- 238000013417 toxicology model Methods 0.000 claims description 5
- 238000011156 evaluation Methods 0.000 claims description 4
- 230000007246 mechanism Effects 0.000 claims description 4
- 230000036228 toxication Effects 0.000 claims description 4
- 229910052751 metal Inorganic materials 0.000 claims description 3
- 239000002184 metal Substances 0.000 claims description 3
- PUKLDDOGISCFCP-JSQCKWNTSA-N 21-Deoxycortisone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@@](C(=O)C)(O)[C@@]1(C)CC2=O PUKLDDOGISCFCP-JSQCKWNTSA-N 0.000 claims description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 2
- FCYKAQOGGFGCMD-UHFFFAOYSA-N Fulvic acid Natural products O1C2=CC(O)=C(O)C(C(O)=O)=C2C(=O)C2=C1CC(C)(O)OC2 FCYKAQOGGFGCMD-UHFFFAOYSA-N 0.000 claims description 2
- 150000001450 anions Chemical class 0.000 claims description 2
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- 238000011109 contamination Methods 0.000 claims description 2
- 239000000284 extract Substances 0.000 claims description 2
- 239000002509 fulvic acid Substances 0.000 claims description 2
- 229940095100 fulvic acid Drugs 0.000 claims description 2
- 230000035784 germination Effects 0.000 claims description 2
- 229910021645 metal ion Inorganic materials 0.000 claims description 2
- 238000011065 in-situ storage Methods 0.000 claims 1
- 238000009938 salting Methods 0.000 abstract description 6
- 238000005516 engineering process Methods 0.000 abstract description 2
- 238000003900 soil pollution Methods 0.000 abstract description 2
- 231100000816 toxic dose Toxicity 0.000 abstract 1
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- 238000004140 cleaning Methods 0.000 description 4
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
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- 230000008901 benefit Effects 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 230000002860 competitive effect Effects 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L magnesium sulphate Substances [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 229910001453 nickel ion Inorganic materials 0.000 description 2
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- 239000011043 treated quartz Substances 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 238000009736 wetting Methods 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 229910018505 Ni—Mg Inorganic materials 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 231100000674 Phytotoxicity Toxicity 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
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- 238000007405 data analysis Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
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- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
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- 231100000614 poison Toxicity 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
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Abstract
The invention discloses a kind of compound prediction techniques and its application to wheat root elongation toxicity in soil of cadmium nickel, belong to ecological heavy metal toxicity assessment technology field.The present invention is in known Cd2+And Ni2+Competing cations be H+And Mg2+On the basis of, consider the osmotic pressure influence of the toxicity inhibition effect and salting liquid of competing cations, the composite toxicity effect of Cd and Ni is calculated using the method that independent toxic concentration is added.The osmotic pressure effect parameter of Cd-Ni composite toxicity effect parameter and salting liquid is obtained using sand culture experiment first, then the parameter of acquisition is applied to the prediction of Cd-Ni composite toxicity effect in different soils.This method is also applied for the compound prediction to other roots of the crop elongation poisonous effect of Cd-Ni, provides new method for heavy metal-polluted soil urban eco landscape forest and regional soil pollution control.
Description
Technical field
The invention belongs to ecological heavy metal toxicity evaluation areas, compound in soil more specifically to a kind of cadmium nickel
Wheat root extends the prediction technique of toxicity, is tested using sand culture and obtains the compound prediction mould to wheat root elongation poisonous effect of cadmium nickel
Type and parameter.This method had both considered the competitive effect of heavy metal ion and other cations and bio-ligand, it is also considered that soil
The parameter of the osmotic pressure effect of earth salting liquid, acquisition has good prediction effect to cadmium nickel composite toxicity in soil type.
Background technique
It is shown according to soil environment quality survey report in 2014, cadmium and nickel are two big heavy metal pollutions of China's most serious
Object.Furthermore nickel-cadmium cell is widely used so that part agricultural land soil is by serious cadmium nickel combined pollution.Cadmium-nickel in environment
Migration with higher can enter food chain by soil-plant system, generate toxic effect to animals and plants.And wheat is made
For one of big cereal in the world three, worldwide plantation extensively, therefore effectively composite toxicity pair of the evaluation cadmium-nickel to wheat
Prevention and treatment heavy metal pollution of soil and human body murder by poisoning are of great significance.
The heavy metal of different shape has different bio-toxicities in soil, and research finds a free state huge sum of money in the soil liquid
The activity of category is directly related with phytotoxicity, and considers that competition of the cation to heavy metal ion coexists in other in the soil liquid
The effect of the osmotic pressure of effect and salting liquid can more accurately describe the poisonous effect of heavy metal on plants.This research is more at present
The description of single heavy metal bio-toxicity is concentrated on, but the bio-toxicity effect of Compound Heavy Metals only can not with single toxicity parameter
Accurate description.Such as the file of Patent No. 201610317197.0 discloses a kind of nickel ion to the pre- of wheat root elongation toxicity
Method and its application are surveyed, invention considers Mg2+Deng cation to the toxicity Competitive assays effect of nickel ion, water a word used in place name condition is established
Lower Ni2+To the determination method of wheat root toxicity.But the invention only has studied the toxicity parameter under nickel Single Pollution, is not suitable for multiple
Pollutional condition is closed, and is invented using there is also differences under solution culture condition, with practical edaphic condition.At present about cadmium in soil
The compound edaphophyte toxicity prediction model of nickel has not been reported.
Summary of the invention
Exist for existing heavy metal in soil methods of Toxicity Assessment and assesses the problems such as not comprehensive, poor for applicability, this hair
It is bright to provide a kind of cadmium nickel compound prediction technique and its application to wheat root elongation toxicity in soil.
Known cadmium and nickel are in the solution Cd there may be the Main Morphology of bio-toxicity2+And Ni2+, it is believed that it is multiple in cadmium nickel
Cd and Ni has similar mechanism of toxication and identical reaction site when closing stress, calculates Cd-Ni using the method that concentration is added
Composite toxicity effect.Consider that the toxicity inhibition effect of cation and the osmotic pressure effect of salting liquid coexist in the soil liquid simultaneously.
It determines Cd-Ni composite toxicity effect parameter and dose-effect equation is to establish cadmium nickel combined pollution in soil to extend wheat root
The key of toxicity prediction method.The present invention is based on bio-ligand toxic models and concentration addition method, using Visual MINTEQ
Cd in the solution that software calculates2+And Ni2+Activity, while considering H+/Mg2+And Cd2+/Ni2+With the competition of bio-ligand, fitting
Obtain cadmium nickel composite toxicity parameter under the conditions of sand culture.In addition, the toxicity that the osmotic pressure for carrying out a large amount of cations extends wheat root
Experiment obtains osmotic pressure effect parameter, and influence of the osmotic pressure to cadmium nickel composite toxicity is corrected by the way of multiplication, establishes and is applicable in
The compound predictive equation to wheat root toxicity of cadmium nickel in soil.Again by the experiment of practical earth culture to the prediction model parameters of acquisition
It is verified, it was demonstrated that the validity of model.
To solve the above-mentioned problems, the technical solution adopted in the present invention is as follows:
A kind of prediction technique of the cadmium nickel compound (Cd-Ni) to wheat root elongation toxicity in soil, comprising the following steps:
(1) composite toxicity is tested:
Breeding is carried out first, selects the consistent wheat seed progress Cd and Ni of germination condition compound and the biology of single stress
Toxicity test.Sand culture experiment is carried out using control variate method.When Cd-Ni Combined Stress, controls other in solution and cation coexists
Concentration is constant, only changes the concentration of Cd and Ni；And when the single stress of Cd/Ni, in known Cd2+And Ni2+Competing cations be H+
And Mg2+On the basis of, only change H in solution+Or Mg2+Concentration and to control other cation concns constant.
(2) Cd-Ni composite toxicity model parameter is determined:
After toxicity test, the root elongation of all processing group wheats is measured, relative toxicity effect value RE% is calculated, calculates
Formula are as follows:
Wherein, RE% indicates relative toxicity effect, LiIndicate that the root of the wheat under Cd-Ni stress extends (cm), LckIt indicates
With LiThe root of the corresponding wheat without Cd-Ni stress extends (cm).
After obtaining relative toxicity effect RE%, it is believed that in composite toxicity effect, Cd and Ni have similar mechanism of toxication,
Occupy identical point during the reaction, i.e. the composite toxicity effect of Cd and Ni are the concentration phase of the independent poisonous effect of Cd/Ni
Add, specific calculation formula are as follows:
Wherein, fCdAnd fNiIndicate the percentage of the point of Cd and Ni in conjunction with bio-ligand in Combined Stress；A and d
For the strength factor of Cd and Ni, coefficient is bigger, and the toxicity for illustrating metal is stronger；β is form parameter.
The competing cations of known Cd and Ni are H+And Mg2+, in conjunction with equation (3) the i.e. calculating of f, equation (2) be can be expressed as:
Wherein, { M2+Be heavy metal cation activity, { X2+Be competing cations activity, { Cd2+}、{Ni2+}、{H+And { Mg2+Indicate solution in each free state ion activity (mol/L), pass through form software for calculation Visual MINTEQ count
It calculates；KCdBL、KHBL、KNiBLAnd KMgBLIndicate the binding constant (L/mol) of each ion and bio-ligand.
(3) binding constant of Cd/Ni and competing cations is determined:
After the single stress toxicity test of Cd/Ni, the root elongation of each processing group wheat is measured, is calculated according to equation (1)
Relative toxicity effect value RE%.Corresponding EC under each cation concn is calculated again50Value, accounting equation are as follows:
Wherein, C indicates alignment processing group metal ion activity { Cd2+Or { Ni2+(mol/L), β is dose-effect equation
Form parameter, the free state ionic activity of each ion passes through form software for calculation Visual MINTEQ and calculates in solution.According to
EC50With H+And Mg2+Relationship, nonlinear fitting is carried out to toxicity tests data using equation (6) and (7), obtains a huge sum of money
The binding constant K of category and competing cations and bio-ligandMBLAnd KXBL。
Wherein, f Cd2+Or Ni2+The percentage of point in conjunction with bio-ligand,M 2+Indicate Cd in solution2+Or Ni2+
Activity (mol/L), {X n+Indicate H+Or Mg2+Activity (mol/L), KMBLAnd KXBLIt is the combination of each cation and bio-ligand
Constant (L/mol).EC50Indicate the Cd in solution when generating 50% biological inhibition effect2+Or Ni2+Activity (mol/L).
(4) the influence experiment of osmotic pressure:
In order to study soil Common Cations (Ca2+、Mg2+、Na+、K+) osmotic pressure to the depression effect of wheat root, carry out
The experiment of osmotic pressure sand culture, i.e., be added without Cd and Ni in experiment, using control variate method, only changes the concentration of some cation simultaneously
It is constant to control other cation concns.Osmotic pressure effect parameter is fitted using following dose-effect equation:
Wherein, I50The ionic strength of solution, β when reaching 50% depression effect for wheat root elongation0For mould shapes parameter.
It is fitted to obtain I by osmotic pressure toxicity test data50And β0。
(5) Cd-Ni composite toxicity model in soil is constructed:
The Cd obtained according to equation (4) and fitting2+/Ni2+And H+/Mg2+With the binding constant K of bio-ligandMBLAnd KXBL,
In conjunction with the osmotic pressure depression effect parameter that osmolarity test obtains, composite toxicity computation model is obtained.It is considered herein that infiltration
Pressing is conspiracy relation between effect and composite toxicity effect, therefore is combined equation (8) and equation (4) by the way of being multiplied,
Obtain the predictive equation of Cd-Ni composite toxicity effect:
(6) earth culture confirmatory experiment method
To verify application effect of the toxicity parameter obtained under sand culture experiment condition in practical soil, this earth culture is devised
Confirmatory experiment.Representative red soil (Kunming, Yunnan) and yellowish soil (Nanjing) are acquired, crosses 2mm after soil is air-dried
Sieve, the contaminated soil of different composite Cd-Ni concentration level is obtained by way of artificially contaminating, keeps soil moisture content after contamination
It is 80% and balance 30 days of field capacity.It is real that exhibition wheat root elongation is decontroled according to the biological toxicity test in claim 1
It tests, growth period uses original position soil solution extractor MicroRhizon (Rhizosphere Research Products, lotus
It is blue) extract the soil liquid, measurement solution ph, total organic carbon TOC, Cd in solution2+、Ni2+, a large amount of cation (K+、Na+、Ca2+、
Mg2+) and anion (Cl-、SO4 2-、NO3 -) concentration, calculate the ionic strength (I) of solution, wherein TOC represents fulvic acid FA and contains
Amount calculates each ionic activity by form software for calculation visual MINTEQ, using cadmium nickel in equation (9) prediction corresponding soils
Composite toxicity effect.
It further, is respectively H in the competing cations of known Cd and Ni+And Mg2+In the case where, using step (3)
The binding constant of determining heavy metal and competing cations and bio-ligand obtains composite toxicity parameter according to equation (4) fitting.
The influence that toxicity is generated using equation (9) Lai Xiuzheng osmotic pressure simultaneously.
Further, the input data of the form software for calculation Visual MINTEQ includes pH value, all ions
With the total concentration of ligand, system is set as open system, i.e., with CO in atmosphere2Balance.
A kind of compound prediction technique heavy metal toxicity in the soil to wheat root elongation toxicity in soil of above-mentioned cadmium nickel
Application in evaluation areas.
A kind of compound prediction technique to wheat root elongation toxicity in soil of above-mentioned cadmium nickel is in wheat planting field
Using.
The compound model ginseng to wheat root elongation poisonous effect of acquisition cadmium nickel is tested using sand culture the present invention provides a kind of
Number, for predicting the prediction technique of cadmium nickel composite toxicity effect in soil, and establishes complete model parameter.
The concentration referred in the present invention is the amount of solute in solution, is indicated with [] or c；And activity refers to effects of ion reality
The effective concentration that border plays a role is indicated with { } or α.Relationship between the two is c=γ α, and wherein γ is activity coefficient.
(7) poisonous effect Computing Principle are as follows:
When calculating cadmium nickel composite toxicity effect, it is believed that Cd and Ni has similar mechanism of toxication, accounts for during the reaction
According to identical point, i.e. Cd is added with the concentration that the composite toxicity effect of Ni is the independent poisonous effect of Cd/Ni.Meanwhile by plant
Root surface reaction site is regarded as bio-ligand (BL), and heavy metal ion can generate poisonous effect in conjunction with bio-ligand and toxicity is imitated
Should the total bio-ligand point of bio-ligand point Zhan in conjunction with by it ratio (f) it is related.Specific formula for calculation are as follows:
Wherein, fCdAnd fNiIndicate the percentage of the point of Cd and Ni in conjunction with bio-ligand in Combined Stress；A and d
For the strength factor of Cd and Ni, coefficient is bigger, and the toxicity for illustrating metal is stronger；β is form parameter.
Common cation (such as K in the soil liquid+、Ca2+、Na+、Mg2+、H+) be considered as to plant nonhazardous effect,
H simultaneously+It can be with Cd2+Compete bio-ligand point and Mg2+Can and Ni2+Bio-ligand point is competed, to mitigate Cd2+With
Ni2+To the bio-toxicity of wheat.Therefore, total point (TBL) of bio-ligand can be expressed as
[TBL]=[BL-]+[MBL+]+[XBL+]
(11)
Wherein, [BL-] indicate the bio-ligand point site concentration not combined by any cation；[MBL+] indicate by Cd2+Or
Ni2+In conjunction with bio-ligand point site concentration；[XBL+] indicate by competing cations H+Or Mg2+In conjunction with bio-ligand point it is dense
Degree,.It is possible thereby to derive Cd2+Or Ni2+The percentage (f) of point in conjunction with bio-ligand:
{M 2+Indicate Cd in solution2+Or Ni2+Activity (mol/L), {X n+Indicate H+Or Mg2+Activity (mol/L), KMBL
And KXBLIt is the binding constant (L/mol) of each cation and bio-ligand.
After obtaining the binding constant of heavy metal and competing cations and BL by equation (12) fitting, equation (12) are substituted into
In equation (10), the calculation formula of composite toxicity effect is obtained:
Wherein, { Cd2+}、{Ni2+}、{H+And { Mg2+Indicate solution in each free state ion activity (mol/L)；KCdBL、
KHBL、KNiBLAnd KMgBLIndicate the binding constant (L/mol) of each ion and bio-ligand.
The root that the osmotic pressure effect that a large amount of cations in the soil liquid can be generated by it influences wheat extends, therefore
Need for osmotic pressure effect to be included in calculating when prediction, after corrected Calculation result, the accounting equation of Cd-Ni composite toxicity are as follows:
Above equation is the models fitting frame of Cd-Ni composite toxicity, model parameter (a, d, I50、β、β0、KMBLWith
KXBL) can be obtained by toxicity test data and Data Analysis Software progress nonlinear fitting, finally establish complete sand culture
Under the conditions of Cd-Ni it is compound to wheat root elongation toxicity prediction method.
Specific technical step are as follows:
Step 1: the sand culture toxicity test of wheat root elongation
Including Cd and Ni be single and the toxicity test of Combined Stress and without the osmotic pressure influence experiment of Cd/Ni stress.In Cd/
When the single stress of Ni, it is known that the competing cations of Cd are H+And the competing cations of Ni are Mg2+.On this basis, using control
Quantity method processed changes H+Or Mg2+Concentration to control other cation concns constant simultaneously.When Cd-Ni Combined Stress, solution is controlled
In other cation concns it is constant, change Cd2+And Ni2+Concentration.In the osmotic pressure influence experiment that development is coerced without Cd/Ni,
K is set+、Na+、Ca2+、Mg2+Four kinds of processing, concentration that every kind of processing changes one of cation control simultaneously other sun from
Sub- concentration is constant.Three repetitions of each processing group.
Before experiment starts, breeding is carried out, after wheat seed disinfection, cleaning, is equably sprinkling upon and is covered with wet filter paper (with super
Pure water wetting) culture dish in.Then culture dish is placed in growth cabinet.Climate box is set as no light, temperature 20
DEG C, air humidity 80%, the wet of filter paper is kept during breeding.Select Germinating status good after 48h and with three about
The seed of 1cm root long is tested.
Sand culture experiment quartz sand particle size used is 0.3-1.0mm, and with 5%HCl to impregnate removal calcium carbonate for 24 hours miscellaneous using preceding
Matter, then clean with pure water the quartz sand of acid processing, until 5.6 or so then to think that quartz sand has cleaned dry for the pH value of cleaning solution
Only, it is baked to spare.Every basin dress 300g treated quartz sand, after corresponding 80mL sand culture solution is mixed with, chooses 8
Satisfactory seed is fixed on quartz sand, then covers 10g quartz sand.Each processing is arranged three in parallel.Experiment is with pure
Water is as blank group, not add Cd or Ni as experimental comparison group, wherein a large amount of cation (K of experimental comparison group+、Ca2+、
Na+、Mg2+) setting it is identical as experimental group.Toxicity test whole process is carried out in growth cabinet, climate box setting are as follows: light application time
16h, 25 DEG C of temperature, air humidity 75%；Interlunation 8h, 20 DEG C of temperature, air humidity 80%.The toxicity test time is
14 days, the moisture to be scattered and disappeared every three days by gravimetric method supplement.Wheat is harvested after 14 days.
Step 2: building composite toxicity prediction model
After growth, the root elongation of all processing group wheats is measured, longest 3 root longs of every plant of wheat is chosen and is surveyed
Amount, the root as one plant of wheat extend situation.The relative toxicity effect of each processing group is calculated using equation (1) according to the root long of measurement
Answer (RE%).
After obtaining RE value, corresponding EC under each cation concn is calculated according still further to equation (5)50It is worth, each ion in solution
Free state ionic activity is calculated by form software for calculation Visual MINTEQ, and software input value includes pH value of solution, all ions
Concentration, Activity Calculation formula use Davies equation, simultaneously because experiment be open system, software calculate when consider atmosphere
In CO2。
According to EC50With H+And Mg2+Relationship, using equation (6) and equation (7) to the toxicity test of the single stress of Cd and Ni
Data carry out nonlinear fitting, obtain the binding constant (K of heavy metal ion and competitive cation and bio-ligandMBLWith
KXBL).The toxicity test data of obtained bio-ligand binding constant and Cd-Ni Combined Stress are combined using equation (13),
It is as shown in table 1 that fitting obtains composite toxicity prediction model parameters (a, d and β).Later according to osmotic pressure influence experiment data into
Row nonlinear fitting obtains osmotic pressure effect parameter (I50And β0) as shown in table 2, model is modified.Finally establish sand culture item
The compound computation model and relevant parameter to wheat root elongation poisonous effect of cadmium nickel under part.
The compound model parameter to wheat root elongation poisonous effect of Cd-Ni in the experiment of 1 sand culture of table
A) unit of K is L/mol；
B) a is the strength factor of Cd, and d is the strength factor of Ni, shows that Cd and Ni extends wheat root in Combined Stress
The power of toxicity.
The model parameter of osmotic pressure effect in the experiment of 2 sand culture of table
a)I50Ionic strength when reaching 50% depression effect for wheat root elongation in solution；
b)β0For the form parameter of osmotic pressure effect model；
Compared with the existing technology, the invention has the benefit that
(1) advantage of the invention is that the Prediction Parameters of the Cd-Ni composite toxicity obtained by sand culture experiment can be direct
For the prediction of Cd-Ni composite toxicity in soil, while experimental method can also be used for the prediction of the root toxicity of other crops.
(2) present invention solve the problems, such as under edaphic condition Cd-Ni it is compound to wheat root toxicity can not Accurate Prediction, examining
Consider feelings of the competing cations on the osmotic pressure of the inhibition influence of Cd and Ni toxicity and cation to wheat root extension effects in solution
Under condition, the prediction technique of Cd-Ni composite toxicity effect under the conditions of sand culture is established.The model parameter of acquisition can be used in soil
The compound prediction to wheat root elongation toxicity of Cd-Ni, mentions for heavy metal-polluted soil urban eco landscape forest and regional soil pollution control
For reference.
(3) method provided by the invention considers the concentration of Cd and Ni, competing cations and osmotic pressure effect etc. in soil
A variety of Marketing Factors, obtained composite toxicity parameter pass through the verifying of different types of soil, are heavy metal in soil plant poison
Property assessment provide science method.
Detailed description of the invention
When Fig. 1 is the single stress of Cd and Ni in the present invention, H+(a) and Mg2+(b) to wheat root toxicity EC50Influence；
Fig. 2 is that (abscissa is effects of ion intensity to cation permeability pressure influence fitted figure, and ordinate is osmotic pressure to small
The opposite depression effect of wheat root elongation)；
Fig. 3 is that (abscissa is practical measurement RE% to composite toxicity effect fitting result figure, and ordinate is mould under the conditions of sand culture
Type predicts RE%, RMSE and R in figure2Root-mean-square error and the coefficient of determination between model predication value and actual observation.Solid line
For 1:1 line, dotted line is the deviation of 1:1 line ± 20%)；
Fig. 4 is the prediction result figure (left side that two kinds of composite toxicity prediction techniques extend toxicity to wheat root in two kinds of soil
Free state Cd in solution is only considered when figure is toxicity prediction2+And Ni2+；Right side is using composite toxicity model parameter of the invention
Prediction result.Wherein, abscissa is the Relative biological toxicity that actual observation arrives, and ordinate is that the model that two methods obtain is pre-
Measured value.RMSE and R in figure2Root-mean-square error and the coefficient of determination between model predication value and actual observation.Solid line is 1:1
Line, dotted line are the deviation of 1:1 line ± 20%).
Specific embodiment
The present invention is further described below combined with specific embodiments below.
Embodiment 1
It is H in the competing cations of known Cd and Ni+And Mg2+On the basis of, carry out the single stress sand culture experiment of Cd/Ni,
Using control variate method, only change H+/Mg2+Concentration while control other cation concentration it is constant.Carry out Cd- again later
The experiment of Ni Combined Stress, controls other cations (K in solution+、Na+、Ca2+、Mg2+) Cd in constant change solution of concentration2+With
Ni2+Concentration.In order to study a large amount of cation (K+、Na+、Ca2+、Mg2+) osmotic pressure influence that wheat root is extended, carry out nothing
The osmotic pressure influence experiment of Cd-Ni stress is only changed the concentration of certain cation while being controlled other sun using control variate method
Ion concentration is constant.There are three repeat for each processing group.The ion component of each processing group sand culture solution is shown in Table 3 and 4.
Before experiment starts, breeding is carried out, after wheat seed disinfection, cleaning, is equably sprinkling upon and is covered with wet filter paper (with super
Pure water wetting) culture dish in.Then culture dish is placed in growth cabinet.Climate box is set as no light, temperature 20
DEG C, air humidity 80%, the wet of filter paper is kept during breeding.Select Germinating status good after 48h and with three about
The seed of 1cm root long is tested.
Sand culture experiment quartz sand particle size used is 0.3-1.0mm, and with 5%HCl to impregnate removal calcium carbonate for 24 hours miscellaneous using preceding
Matter, then clean with pure water the quartz sand of acid processing, until 5.6 or so then to think that quartz sand has cleaned dry for the pH value of cleaning solution
Only, it is baked to spare.Every basin dress 300g treated quartz sand, after corresponding 80mL sand culture solution is mixed with, chooses 8
Satisfactory seed is fixed on quartz sand, then covers 10g quartz sand.Each processing is arranged three in parallel.Experiment is with pure
Water is as blank group, not add Cd or Ni as experimental comparison group, wherein a large amount of cation (K of experimental comparison group+、Ca2+、
Na+、Mg2+) setting it is identical as experimental group.Toxicity test whole process is carried out in growth cabinet, climate box setting are as follows: light application time
16h, 25 DEG C of temperature, air humidity 75%；Interlunation 8h, 20 DEG C of temperature, air humidity 80%.The toxicity test time is
14 days, the moisture to be scattered and disappeared every three days by gravimetric method supplement.Wheat is harvested after 14 days.
3 sand culture of table tests each processing group concentration setting
Each processing group concentration setting in the experiment of 4 osmotic pressure effect sand culture of table
After growth, the root elongation of all processing group wheats is measured, longest 3 root longs of every plant of wheat is chosen and is surveyed
Amount, the root as one plant of wheat extend situation.Cd or Ni is calculated to the opposite of wheat root using equation (1) according to the root long of measurement
Poisonous effect (RE%).
After obtaining RE value, corresponding EC under each cation concn is calculated according still further to equation (5)50It is worth, each ion in solution
Free state ionic activity is calculated by form software for calculation Visual MINTEQ, and software input value includes pH value of solution, all ions
Concentration, Activity Calculation formula use Davies equation, simultaneously because experiment be open system, software calculate when consider atmosphere
In CO2.The poison relatively of the single stress of the Cd/Ni being calculated, Cd-Ni Combined Stress and salting liquid osmolarity test processing group
Property effect RE%, EC50With β value as shown in table 5,6 and 7.
The relevant parameter of each processing group of the single stress experiment of table 5 Cd and Ni
Corresponding a.Cd-H group is pH value, and corresponding Ni-Mg group is MgSO4Concentration, unit: mM
6 Cd-Ni Combined Stress of table tests the relevant parameter of each processing group
It is the concentration of Cd and Ni that a.Cd-Ni group is corresponding, and unit is μM；
B.a is the strength factor of Cd, and d is the strength factor of Ni, and the toxicity for respectively indicating Cd and Ni is strong and weak.
The relevant parameter of each processing group of 7 osmolarity test of table
a.NaCl、CaCl2、KCl、MgSO4Corresponding group is concentration, unit: mM
Wheat (Yangmai 13) is after breeding, sand culture experiment in 14 days, to EC50And H+/Mg2+Carry out correlation analysis (figure
1), competing cations and Cd/Ni root toxicity EC50Linear equation are as follows:
EC50(Cd)=389 × { H+}+26.2
EC50(Ni)=52.8 × { Mg2+}+77.4
(15)
Wherein, EC50Unit be μM, H+And Mg2+The unit of activity is mM.
The free state activity that each ion in each processing group solution is calculated using Visual MINTEQ, using equation (6)
(7) nonlinear fitting is carried out to sand culture experimental data using Data Analysis Software, fitting result finds { Cd2+}、{Ni2+And it is big
Amount cation permeability pressure (Fig. 2) shows apparent poisonous effect to wheat root elongation.Fitting obtain Cd/Ni and competition sun from
Son (H+/Mg2+) and biological binding constant KMBLAnd KXBLAnd osmotic pressure effect parameter (I50And β0).Cd- is combined according to equation (4)
Ni composite toxicity data, fitting obtain composite toxicity prediction model parameters (a, d and β), as a result as shown in Figure 3.Specific model
Parameter is as shown in Table 1 and Table 2.
For the accuracy for verifying proposition method of the present invention, parameter that serial earth culture experimental verification is fitted is carried out in reality
Usable condition in the soil potted plant experiment of border.It acquires representative Kunming, Yunnan red soil and Nanjing yellowish soil and carries out soil
Training experiment, physiochemical properties of soil are as shown in table 8.Verification result is as shown in figure 4, illustrate the prediction of this method and model parameter
Effect is preferable, and prediction result is much better than free state Cd in only consideration solution2+And Ni2+The prediction result of activity.
8. physiochemical properties of soil of table
Claims (6)
1. a kind of compound prediction technique to wheat root elongation toxicity in soil of cadmium nickel, which comprises the following steps:
(1) composite toxicity is tested:
Breeding is carried out first, selects the consistent wheat seed progress Cd and Ni of germination condition compound and the bio-toxicity of single stress
Experiment carries out sand culture experiment using control variate method, when Cd-Ni Combined Stress, controls other in solution and cation concn coexists
It is constant, only change the concentration of Cd and Ni；And when the single stress of Cd/Ni, in known Cd2+And Ni2+Competing cations be H+And Mg2 +On the basis of, only change H in solution+Or Mg2+Concentration and to control other cation concns constant；
(2) Cd-Ni composite toxicity model parameter is determined:
After toxicity test, the root elongation of measurement all processing group wheats of Combined Stress calculates relative toxicity effect value RE%,
Calculation formula are as follows:
Wherein, RE% indicates relative toxicity effect, LiIndicate that the root of the wheat under Cd-Ni stress extends (cm), LckExpression and LiIt is right
The root elongation (cm) for the wheat without Cd-Ni stress answered；
After obtaining relative toxicity effect RE%, in composite toxicity effect, Cd and Ni have similar mechanism of toxication, are reacting
Identical point is occupied in journey, i.e. composite toxicity effect is that Cd is added with the concentration of the independent poisonous effect of Ni, specific to calculate public affairs
Formula are as follows:
Wherein, fCdAnd fNiIndicate the percentage of the point of Cd and Ni in conjunction with bio-ligand in Combined Stress；A and d be Cd and
The strength factor of Ni, coefficient is bigger, and the toxicity for illustrating metal is stronger；β is form parameter；
It is H in the competing cations of known Cd and Ni+And Mg2+On the basis of, in conjunction with equation (3) the i.e. calculating of f, equation (2) can
Expression are as follows:
Wherein, { M2+Be heavy metal cation activity, { X2+Be competing cations activity, { Cd2+}、{Ni2+}、{H+And
{Mg2+Indicate solution in each free state ion activity (mol/L)；KCdBL、KHBL、KNiBLAnd KMgBLIndicate that each ion is matched with biology
The binding constant (L/mol) of body；
(3) binding constant of Cd/Ni and competing cations is determined:
After the single stress toxicity test of Cd/Ni, the root elongation of each processing group wheat is measured, is calculated according to equation (1) opposite
Poisonous effect value RE%；Corresponding EC under each cation concn is calculated again50Value, accounting equation are as follows:
Wherein, C indicates alignment processing group metal ion activity { Cd2+Or { Ni2+(mol/L), β is the shape of dose-effect equation
Shape parameter, the free state ionic activity of each ion passes through form software for calculation Visual MINTEQ and calculates in solution；According to EC50
With H+And Mg2+Relationship, nonlinear fitting is carried out to toxicity tests data using equation (6) and (7), obtain heavy metal and
The binding constant K of competing cations and bio-ligandMBLAnd KXBL；
Wherein, f Cd2+Or Ni2+The percentage of point in conjunction with bio-ligand, { M2+Indicate Cd in solution2+Or Ni2+Work
It spends (mol/L), { Xn+Indicate competing cations H+Or Mg2+Activity (mol/L), KMBLAnd KXBLIt is each cation and bio-ligand
Binding constant (L/mol)；EC50Indicate the Cd in solution when generating 50% biological inhibition effect2+Or Ni2+Activity (mol/
L)；
(4) the influence experiment of osmotic pressure:
In order to study Common Cations In Soil (Ca2+、Mg2+、Na+、K+) osmotic pressure to the depression effect of wheat root, carry out and seep
Pressure sand culture experiment thoroughly, i.e., be added without Cd and Ni in experiment, using control variate method, only changes the same time control of concentration of some cation
It is constant to make other cation concns；Osmotic pressure effect parameter is fitted using following dose-effect equation:
Wherein, I is the ionic strength of solution, I50The ionic strength of solution, β when reaching 50% depression effect for wheat root elongation0
For mould shapes parameter, it is fitted to obtain I by osmotic pressure toxicity test data50And β0；
(5) Cd-Ni composite toxicity model in soil is constructed:
The Cd obtained according to equation (4) and fitting2+/Ni2+And H+/Mg2+With the binding constant K of bio-ligandMBLAnd KXBL, in conjunction with
The osmotic pressure depression effect parameter that osmolarity test obtains, obtains the computation model of Cd-Ni composite toxicity effect under edaphic condition；
Due to being conspiracy relation between osmotic pressure effect and composite toxicity effect, by equation (8) and equation by the way of being multiplied
(4) it combines, obtains the predictive equation of Cd-Ni composite toxicity effect:
2. a kind of compound prediction technique to wheat root elongation toxicity in soil of cadmium nickel according to claim 1, feature
It is: is respectively H in the competing cations of known Cd and Ni+And Mg2+In the case where, using step (3) determine heavy metal and
The binding constant of competing cations and bio-ligand obtains composite toxicity parameter according to equation (4) fitting；Equation is used simultaneously
(9) influence that Lai Xiuzheng osmotic pressure generates toxicity.
3. a kind of compound prediction technique to wheat root elongation toxicity in soil of cadmium nickel according to claim 1, feature
It is: Cd in solution is calculated using form software for calculation Visual MINTEQ2+And Ni2+Activity；Software input value includes molten
The concentration of liquid pH, all ions, Activity Calculation formula use Davies equation, while it is open system that system, which is arranged, that is, is considered
CO in atmosphere2。
4. a kind of compound prediction technique to wheat root elongation toxicity in soil of cadmium nickel described in claim 1 is in heavy metal toxicity
Application in evaluation areas.
5. a kind of compound prediction technique to wheat root elongation toxicity in soil of cadmium nickel described in claim 1 is led in wheat planting
Application in domain.
6. a kind of compound side to the prediction technique of wheat root elongation toxicity in soil of cadmium nickel described in verifying claim 1
Method, it is characterised in that: acquire representative red soil and yellowish soil, 2mm sieve is crossed after soil is air-dried, passes through what is artificially contaminated
Mode obtains the contaminated soil of different composite Cd-Ni concentration level, and it is field capacity that soil moisture content is kept after contamination
80% and balance 30 days；Exhibition wheat root elongation experiment is decontroled according to the biological toxicity test, growth period uses in-situ soil
Earth solution extractor MicroRhizon extracts the soil liquid, measurement solution ph, total organic carbon TOC, Cd in solution2+、Ni2+、
The concentration of a large amount of cations and anion, calculates the ionic strength of solution, wherein TOC represents fulvic acid FA content, passes through form
Software for calculation Visual MINTEQ calculates each ionic activity, is imitated using cadmium nickel composite toxicity in equation (9) prediction different soils
It answers.
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