CN113607909A - Method for constructing water body comprehensive toxicity characterization parameters by algae photosynthetic inhibition method - Google Patents
Method for constructing water body comprehensive toxicity characterization parameters by algae photosynthetic inhibition method Download PDFInfo
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Abstract
The invention provides a method for constructing water body comprehensive toxicity characterization parameters by an algae photosynthetic inhibition method, which comprises the following steps: step 1: measuring chlorophyll fluorescence rising kinetic curve OJIP and obtaining F0、FJ、FI、FmPhotosynthetic fluorescence parameters; step 2: to FJ、FICarrying out normalization processing to obtain two characteristic site data items FJ *、FI *Extracting information of J point and I point in the curve, and eliminating the influence of fluorescence measurement absolute intensity; and step 3: respectively calculating OJIP curve at [ T ] by using fixed integralF0,TFJ]、[TF0,TFI]、[TF0，TFp]In the interval with F ═ F0Area S surrounding the closed regionJ、SI、SP(ii) a And 4, step 4: by using SPTo SJ、SICarrying out normalization processing to obtain two curve segment data items SJ *、SI *Extracting the rising process information of O-J and J-I section curves, and step 5: according to FJ *、FI *、SJ *、SI *And TF0、TFJ、TFI、TFJ‑TF0、TFI‑TF0Calculating the comprehensive characterization parameter PICTE(ii) a Step 6: respectively calculating PI of an actual water body experimental group and a blank control groupCTEAnd comparing the two to calculate the change rate of the comprehensive toxicity parameter, and judging the toxicity intensity of the water body substance to be detected.
Description
Technical Field
The invention belongs to the field of resources and environment, and particularly relates to a method for constructing water body comprehensive toxicity characterization parameters by an algae photosynthetic inhibition method.
Background
Along with the aggravation of human activities in recent years, more and more pollutants are directly or indirectly discharged into environmental water bodies to cause great impact on aquatic ecosystems, and the rapid detection of water quality toxicity has important significance for preventing and treating water body pollution and guaranteeing water quality safety. The conventional water toxicity detection methods include a rational analysis method, a biotoxicity detection method and the like. The biological toxicity detection is widely used because the comprehensive toxicity of the polluted water body to the biological population can be intuitively reflected due to the interaction of various toxic substances. The biotoxicity detection method based on the algae photosynthesis inhibition effect has the characteristics of rapidness, convenience, real-time online detection and the like, and is widely applied to water toxicity detection in recent years.
The biotoxicity detection method based on the algae photosynthetic inhibition effect takes chlorophyll fluorescence in vivo as a natural probe, and determines the toxic effect of toxic substances in water by measuring the change of the algae photosynthetic fluorescence parameters, wherein the photosynthetic fluorescence parameters reflect the photosynthetic metabolism state of algae in real time and can be used for rapidly detecting the water toxicity. At present, the maximum photochemical quantum yield Fv/Fm is mostly adopted as the photosynthetic fluorescence parameter for toxicity detection in the prior research. Such as Juneau, etc., to investigate the Hg pairs of four kinds of algae, namely Fusarium cellulosum, crescent moon algae, Chlorella and microcystis2+The study suggested that Fv/Fm is Hg2+Appropriate indicators for toxicity testing; skihi et al showed that Fv/Fm can be used to assess glyphosate toxicity. However, the action sites and mechanisms of different kinds of water body pollutants on algae photosynthesis inhibition are different, the photosynthetic fluorescence parameters which cause response to the different kinds of water body pollutants are also different, and a single photosynthetic fluorescence parameter Fv/Fm is not responsive to certain pollutants, so that the detectable pollutant kinds are fewer. In addition, the characterization parameter Fv/Fm for detecting the comprehensive toxicity of the water body by the current algae photosynthesis inhibition method can only reflect the photochemical information of PS II under dark adaptation of photosynthesis, and has low detection sensitivity and limited types of toxic substances.
Disclosure of Invention
Aiming at solving the technical problems that in the water body pollutant toxicity detection research, the photosynthetic fluorescence parameters are more, the single fluorescence parameter Fv/Fm water body toxicity detection sensitivity is not high, the detectable pollutant types are less and the like, the invention provides a photosynthetic inhibition effect characterization parameter PI based on multi-parameter fusionCTEThe method of (1). The method takes rapid chlorophyll fluorescence OJIP as a research object, fuses J, I-point fluorescence information by analyzing response difference of fluorescence of different characteristic points to toxic substances, and provides that J, I-point occurrence time and curve enclosed area are adopted to carry out data item weighting to solve the problem of detecting pollutant types in water toxicity detectionLess and low sensitivity, and provides an effective means for rapid and accurate measurement of comprehensive toxicity of water.
The technical scheme of the invention is as follows: a method for constructing water body comprehensive toxicity characterization parameters by an algae photosynthetic inhibition method comprises the following steps:
step 1: measuring chlorophyll fluorescence rising kinetic curve OJIP and obtaining F0、FJ、FI、FmPhotosynthetic fluorescence parameters;
step 2: using the maximum and minimum normalization method to pair FJ、FICarrying out normalization processing to obtain two characteristic site data items FJ *、FI *Extracting information of J point and I point in the curve, and eliminating the influence of fluorescence measurement absolute intensity;
and step 3: respectively calculating OJIP curve at [ T ] by using fixed integralF0,TFJ]、[TF0,TFI]、[TF0,TFP]In the interval with F ═ F0Area S surrounding the closed regionJ、SI、SP；
And 4, step 4: by using SPTo SJ、SICarrying out normalization processing to obtain two curve segment data items SJ *、SI *Extracting the rising process information of O-J and J-I section curves, and eliminating the influence of the absolute intensity of curve measurement;
and 5: according to FJ *、FI *、SJ *、SI *Calculation result and TF0、TFJ、TFI、TFJ-TF0、TFI-TF0Calculating by using a formula (1) to obtain a comprehensive characterization parameter PICTE；
Wherein, FJ *、FI *The fluorescence intensity of J, I points in the normalized OJIP curve, TF0、TFJ、TFI、TFpRespectively corresponding to O, J, I, P points, SJ *、SI *The normalized O-J, O-I section curve and F (F0) form the area of a closed area respectively;
step 6: respectively calculating the comprehensive toxicity parameters PI of the actual water body experimental group and the blank control group according to the steps 1-5’ CTEAnd PICTEAnd calculating the comprehensive toxicity parameter change rate eta, and judging the toxicity intensity of the water body substance to be detected.
Has the advantages that:
1. the invention utilizes the biological principle that the electron transfer process of algae photosynthesis is blocked when toxic substances stress, thereby causing the change of the fluorescence intensity of the characteristic site in an OJIP curve and the curve ascending process, and utilizes the response difference between the J, I characteristic site in the OJIP curve and the O-J, J-I curve ascending process and the toxic substances to screen the photosynthetic fluorescence parameter F capable of powerfully representing the toxicity of environmental toxicantsJ、FI、SJ、SI。
2. The invention utilizes the maximum and minimum normalization method to FJ、FICarrying out normalization processing to obtain a characteristic site data item FJ *、FI *Will FJ、FIThe value range is also scaled to [0,1 ]]Within the interval, eliminate FJ、FIAffected by the absolute intensity of the fluorescence measurement.
3. The invention calculates the relative area of the enclosed area of the O-J, O-I section curve under the maximum enclosed area by utilizing SPTo SJ、SICarrying out normalization processing to obtain two curve segment data items SJ *、SI *Extracting the rising process information of O-J and J-I curves and eliminating SJ、SIAffected by the absolute intensity of the curve measurement.
4. The invention adopts the number of each characteristic pointForm S of multiplication of data items by data items of curve segments and accumulationJ *×FJ *+SI *×FI *The characteristic that different photosynthetic fluorescence parameters have different responses to toxic substances is fully exerted, and the range of detecting the types of the toxic substances by the photosynthetic fluorescence parameters is expanded.
5. The invention utilizes the appearance time T of J, I characteristic points in the OJIP curveFJ、TFIFor feature location data item FJ *、FI *Making correction by using O-J and J-I section curve process time period length TFJ-TF0、TFI-TF0For two curve segment data items SJ *And SI *And (4) correcting to improve the stability and sensitivity of detection of the toxicity of the substance.
6. According to the biological principle that the electron transfer process of algae photosynthesis is blocked when toxic substances are stressed, so that the fluorescence intensity of characteristic sites in an OJIP curve and the curve ascending process change, the comprehensive toxicity parameters PI of an actual water body experimental group and a blank control group are respectively calculated’ CTEAnd PICTEAnd calculating the comprehensive toxicity parameter change rate eta so as to judge the toxicity strength of the water body substance to be detected.
Drawings
FIG. 1: ojis curve and associated fluorescence parameters;
FIG. 2(a) Fv/Fm, PI under stress of diuron at various concentrationsCTEA response trend graph;
FIG. 2(b) Fv/Fm, PI under stress of different concentrations of 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinoneCTEA response trend graph;
FIG. 2(c) Fv/Fm, PI under the stress of methyl viologen at different concentrationsCTEA response trend graph;
FIG. 2(d) shows Fv/Fm, PI under stress of malathion at various concentrationsCTEA response trend graph;
FIG. 2(e) Fv/Fm, PI under varying concentration of carbofuran stressCTEAnd (5) responding to the trend graph.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, rather than all embodiments, and all other embodiments obtained by a person skilled in the art based on the embodiments of the present invention belong to the protection scope of the present invention without creative efforts.
According to the embodiment of the invention, the response characteristics of O, J, I, P characteristic sites in OJIP to toxic substances are analyzed, and the characteristic parameter PI of the photosynthetic inhibitory effect based on multi-parameter fusion is providedCTEThe method of (1). The method is selected from FJ、FI、SJ、SIBased on the method, J, I point fluorescence intensity information and O-J, J-I curve ascending process information are fused in a summing mode through normalization processing, meanwhile, weights are distributed to each point data item according to the difference of J, I points on response sensitivity of toxic substances by utilizing a weighting thought, and a characterization parameter PI reflecting algae photosynthetic inhibition effect is constructedCTE。
According to one embodiment of the invention, the chlorella vulgaris is taken as a tested alga species, and Fv/Fm, PI and Fv/Fm stress and stress of five common water pollutants of diuron, 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone, methyl viologen, malathion and carbofuran are researchedCTEPerformance difference of two photosynthetic fluorescence parameters in detecting toxic substances, and verification of PICTEThe effectiveness of the parameters in toxicity testing. The method specifically comprises the following steps:
step 1: measuring chlorophyll fluorescence rising kinetic curve OJIP and obtaining F0、FJ、FI、FmPhotosynthetic fluorescence parameters;
step 2: using the maximum and minimum normalization method to pair FJ、FICarrying out normalization processing to obtain two characteristic site data items FJ *、FI *Extracting information of J point and I point in the curve, and eliminating the influence of fluorescence measurement absolute intensity;
and step 3: respectively calculating OJIP curve at [ T ] by using fixed integralF0,TFJ]、[TF0,TFI]、[TF0,TFP]In the interval with F ═ F0Area S surrounding the closed regionJ、SI、SP(ii) a Namely SJIs O, J, T in the OJIP curveF0、TFJThe enclosed area of the enclosed area formed by four points, SIIs O, I, T in the OJIP curveF0、TFIThe enclosed area of the enclosed area formed by four points, SpIs O, I, T in the OJIP curveF0、TFpThe enclosed area of the enclosed area formed by the four points;
and 4, step 4: by using SPTo SJ、SICarrying out normalization processing to obtain two curve segment data items SJ *、SI *Extracting the rising process information of O-J and J-I section curves, and eliminating the influence of the absolute intensity of curve measurement;
and 5: according to FJ *、FI *、SJ *、SI *Calculation result and TF0、TFJ、TFI、TFJ-TF0、TFI-TF0Calculating by using a formula (1) to obtain a comprehensive characterization parameter PICTE；
Wherein, FJ *、FI *The fluorescence intensity of J, I points in the normalized OJIP curve, TF0、TFJ、TFI、TFpRespectively corresponding to O, J, I, P points, SJ *、SI *The normalized O-J, O-I section curve and F (F0) form the area of a closed area respectively;
step 6: respectively calculating the comprehensive toxicity parameters PI of the actual water body experimental group and the blank control group according to the steps 1-5’ CTEAnd PICTECalculating the change rate eta of the comprehensive toxicity parameter, and judging the toxicity intensity of the water body substance to be detectedThe larger the change rate is, the stronger the comprehensive toxicity of the water body is.
Specifically, step 1, measuring chlorophyll fluorescence rising kinetic curve OJIP and obtaining F0、FJ、FI、FmPhotosynthetic fluorescence parameters; wherein, F0、FJ、FI、FmThe acquisition is specifically as follows:
chlorophyll fluorescence rise kinetic curve OJIP was measured and fluorescence data from 20. mu.s-2 s was recorded. As shown in Table 1, F was obtained by calculating the fluorescence values at different times on the OJIP curve0、FJ、FI、Fm。
TABLE 1 basic description of F0, FJ, FI, Fm parameters
Specifically, the step 2: using the maximum and minimum normalization method to pair FJ、FICarrying out normalization processing to obtain two characteristic site data items FJ *、FI *Extracting information of J point and I point in the curve, and eliminating the influence of fluorescence measurement absolute intensity; wherein, FJ、FIThe normalization process is as follows:
scaling original J, I point fluorescence in equal proportion by adopting a min-max normalization method (shown as a formula (2)), wherein X in the formula (2) is data to be normalized, min and max are respectively the minimum value and the maximum value of the value range of the parameter X, and X is the value range of the parameter X*And the values are normalized by X. In the complete OJIP curve, the minimum value is F0Maximum value of FmNormalized J, I point fluorescenceThe values are shown in equations (3) and (4), respectively.
Step 3, respectively calculating the OJIP curve at [ T ] by utilizing fixed integralF0,TFJ]、[TF0,TFI]、[TF0,TFP]In the interval with F ═ F0Area S surrounding the closed regionJ、SI、SP；
The step 4: by using SPTo SJ、SICarrying out normalization processing to obtain two curve segment data items SJ *、SI *Extracting the rising process information of O-J and J-I section curves, and eliminating the influence of the absolute intensity of curve measurement;
the O-J, O-I section OJIP curve and F0 form the area S of the closed areaJ、SIThe maximum area enclosed by the oji curve and F-F0, i.e. the enclosed area S of the O-P curvePFurther calculating the relative area of the enclosed area of the curve of the O-J, O-I section under the maximum enclosed area; finally using SPTo SJ、SICarrying out normalization processing to obtain two curve segment data items SJ *、SI *Extracting the ascending process information of O-J and J-I section curves and eliminating the influence of the curve measurement absolute intensity, wherein SJ、SI、Sp、SJ *、SI *The calculation method is shown in formulas (5), (6), (7), (8) and (9):
and 5: according to FJ *、FI *、SJ *、SI *Calculation result and TF0、TFJ、TFI、TFJ-TF0、TFI-TF0Calculating by using a formula (1) to obtain a comprehensive characterization parameter PICTE(ii) a The method comprises the following specific steps:
pair J, I feature point information FJ *、FI *And O-J and J-I section curve process information SJ *、SI *And fusion, wherein the fusion method adopts the mode that each characteristic point data item is multiplied by a curve section data item and then accumulated: sJ *×FJ *+SI *×FI *. On the basis of the above-mentioned data item F of every characteristic positionJ *、FI *Divided by respective times of occurrence TFJ、TFIJ, I, correcting the feature point data item; two curve segment data items SJ *、SI *Divided by the respective corresponding time interval length TFJ-TF0、TFI-TF0On the O-J and J-I curve segment data itemsCorrecting to construct a water body comprehensive toxicity characterization parameter PICTE：
According to the embodiment of the invention, the method is used for detecting the toxicity of the water body.
Respectively calculating comprehensive toxicity parameters PI of the water body experimental group and the blank control group according to the formula (1)’ CTEAnd PICTEAnd (3) calculating the comprehensive water toxicity parameter change rate eta according to the formula (2), judging the toxicity intensity of the water substance to be detected, establishing a dose-effect relation curve of toxicity response according to the comprehensive toxicity parameter change rate eta and the concentration of the toxic substance, and realizing the quantitative analysis of the toxicity intensity of the water pollutants.
The chlorella vulgaris is taken as a tested alga species, and Fv/Fm and PI are researched under the stress of five common water pollutants of diuron, 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone, methyl viologen, malathion and carbofuranCTEPerformance difference of two photosynthetic fluorescence parameters in detecting toxic substances, and verification of PICTEThe effectiveness of the parameters in toxicity testing.
According to the embodiment of the invention, the method is compared with the traditional method, and comprises a detection range, a lowest detection limit and a highest detection limit; the method comprises the following specific steps:
1. detection range
According to the experimental results of the chlorella vulgaris under the stress of five toxic substances, Fv/Fm and PI are respectively calculated by utilizing a one-way ANOVA methodCTEThe two photosynthetic fluorescence parameters were compared for their ability to detect five toxic substances. Significant differences (P) were observed between the fluorescence parameters of the control and experimental groups above a certain test concentration in the range of the test concentration of the toxic substance (P)<0.05), indicating that the fluorescence parameter has the ability to detect this substance, the results are shown in table 2.
TABLE 2 Fv/Fm, PIABS、PICTEResponse to five toxic substancesResults of differential analysis with control group
Note: the differential values between the control and experimental fluorescence parameters at the 0.05 and 0.01 levels are indicated.
As is clear from Table 2, diuron, 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone, methyl viologen-pair Fv/Fm, and PICTEThe effect is obvious, and the two fluorescence parameters have obvious changes (P) compared with the control group at the concentrations of 1 mug/L, 100 mug/L and 5mg/L respectively<0.05); malathion and carbofuran only show the effect on PICTEThe significant effect of the compound is that the compound shows significant difference at the concentration of 5 mu g/L, and the results of the experimental group and the control group of Fv/Fm have no significant difference under the stress of malathion and carbofuran with different concentrations (P)>0.05), the result shows that the Fv/Fm does not have the capability of detecting malathion and carbofuran, and the Fv/Fm and PI have the capability of detecting toxic substancesCTELess than the detectable contaminant species.
2. Minimum limit of detection
The experiment is carried out under the stress of low concentration of five toxic substances, and Fv/Fm and PI are respectively calculatedCTEThe inhibition/promotion rate, the dose-response curve was established. The lowest detection limit of each fluorescence parameter for toxic substances was calculated using a linear dose-effect curve at low concentrations, according to the International Union of Pure and Applied Chemistry (IUPAC) definition of the lowest detection limit (see equation 8).
Wherein S isbThe standard deviation of the response signal was measured for a number of times for the blank (blank was measured 20 times in this study); m is the slope of the linear dose response curveAnd (4) rate.
TABLE 3 Fv/Fm, PICTEMinimum detection limit for five toxic substances
Note: (-) represents the parameter as non-responsive to toxic substances.
As shown in Table 3, PICTEHas lower detection limit on five toxic substances and stronger detection capability on low-concentration toxic substances, such as Fv/Fm and PICTEThe lowest detection limit of diuron is 2.70 and 0.90 mu g/L respectively, and PICTEThe detection limit of (A) is significantly lower than that of Fv/Fm, and is reduced by 66.7 percent compared with that of Fv/Fm.
3. Maximum limit of detection
According to the experimental results of five toxic substances under high concentration stress, Fv/Fm and PI are respectively establishedCTEIn the toxic substance testing concentration range, when the toxic substance testing concentration is higher than a certain testing concentration, the change rate of the toxicity characterizing parameter reaches a threshold value or tends to be stable, and the testing concentration is the upper limit of the parameter detection.
Fv/Fm、PICTEThe dose-response curves under the stress of the five toxic substances are shown in fig. 2, and the upper detection limits of the two photosynthetic fluorescence parameters on the five toxic substances are respectively calculated according to the dose-response trends, for example, when the concentration of 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone is less than 5mg/L, the Fv/Fm change rate gradually increases with the increase of the concentration of 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone, and when the concentration of 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone is greater than 20mg/L, the Fv/Fm tends to be stable. Therefore, the highest response concentration of the Fv/F is 20mg/L for 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone. And for PICTEWhen the concentration of 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone is in the range of 1 to 40mg/L, PICTEThe rate of change increased with increasing concentration of 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone, and when the concentration of 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone reached the maximum value of 40mg/L of the assay concentration, PI was added to the assay concentrationCTEThe rate of change does not reach the threshold orTends to be smooth and therefore takes PICTEThe highest response concentration of the derivative to the 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone is greater than 40mg/L as a response index, and compared with Fv/Fm, PI (polyimide) is addedCTEThe highest response concentration of the 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone is higher.
According to the same analysis method, Fv/Fm and PI are usedCTEThe maximum response concentrations of five toxic substances of diuron, 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone, methyl viologen, malathion and carbofuran which take two photosynthetic fluorescence parameters as response indexes are listed in Table 4, and PI is viewed from the wholeCTEThe detection upper limit of the five toxic substances is highest, the detection capability is all mg/L, and the detection capability of the five toxic substances is strongest.
TABLE 4 detection upper limits of Fv/Fm, PIABS, PICTE for five toxic substances
In conclusion, the results show that: in the aspect of toxicity detection, the constructed PI is used in the inventionCTEAs a response index, the kit is sensitive to the response of five toxic substances, and can detect more kinds of pollutants such as malathion, carbofuran and the like compared with Fv/Fm; PI (proportional integral)CTEThe minimum detection limits of diuron, 2, 5-dibromo-6-isopropyl-3-methyl-1, 4-benzoquinone and methyl viologen are respectively reduced by 66.7%, 14.4% and 22.9% compared with Fv/Fm, and the detection capability of the reagent on high-concentration toxic substances is stronger. The research provides important toxicity characterization parameters for researching a high-sensitivity and high-precision water body comprehensive toxicity detection method.
Although illustrative embodiments of the present invention have been described above to facilitate the understanding of the present invention by those skilled in the art, it should be understood that the present invention is not limited to the scope of the embodiments, but various changes may be apparent to those skilled in the art, and it is intended that all inventive concepts utilizing the inventive concepts set forth herein be protected without departing from the spirit and scope of the present invention as defined and limited by the appended claims.
Claims (7)
1. A method for constructing water body comprehensive toxicity characterization parameters by an algae photosynthetic inhibition method is characterized by comprising the following steps:
step 1: measuring chlorophyll fluorescence rising kinetic curve OJIP and obtaining F0、FJ、FI、FmPhotosynthetic fluorescence parameters;
step 2: using the maximum and minimum normalization method to pair FJ、FICarrying out normalization processing to obtain two characteristic site data items FJ *、FI *Extracting information of J point and I point in the curve, and eliminating the influence of fluorescence measurement absolute intensity;
and step 3: respectively calculating OJIP curve at [ T ] by using fixed integralF0,TFJ]、[TF0,TFI]、[TF0,TFP]In the interval with F ═ F0Area S surrounding the closed regionJ、SI、SP；
And 4, step 4: by using SPTo SJ、SICarrying out normalization processing to obtain two curve segment data items SJ *、SI *Extracting the rising process information of O-J and J-I section curves, and eliminating the influence of the absolute intensity of curve measurement;
and 5: according to FJ *、FI *、SJ *、SI *Calculation result and TF0、TFJ、TFI、TFJ-TF0、TFI-TF0Calculating by using a formula (1) to obtain a comprehensive characterization parameter PICTE；
Wherein, FJ *、FI *The fluorescence intensity of J, I points in the normalized OJIP curve, TF0、TFJ、TFI、TFPRespectively corresponding to O, J, I, P points, SJ *、SI *The normalized O-J, O-I section curve and F (F0) form the area of a closed area respectively;
step 6: respectively calculating the PI of the actual water body experimental group and the blank control group according to the steps 1-5CTEComparing the two PICTEAnd judging the toxicity strength of the water body substances to be detected according to the difference between the two.
2. The method for constructing the comprehensive water toxicity characterization parameters by the algae photosynthesis inhibition method according to claim 1, wherein in the step 1, a chlorophyll fluorescence rising kinetic curve OJIP is measured, fluorescence data is recorded according to time, and F is obtained by calculating fluorescence values at different times on the OJIP curve0、FJ、FI、Fm。
3. The method for constructing water body comprehensive toxicity characterization parameters by algae photosynthesis inhibition method according to claim 1, wherein in the step 2, the original J, I point fluorescence is scaled in equal proportion by adopting a maximum and minimum normalization method to eliminate the influence of fluorescence measurement absolute intensity, X in the formula (2) is data to be normalized, min and max are respectively the minimum value and the maximum value of the value range of the parameter X, and X is the minimum value and the maximum value of the value range of the parameter X*The minimum value is F in the complete OJIP curve for the value after X normalization0Maximum value of FmThe normalized fluorescence values at J, I points are shown in equations (3) and (4), respectively:
4. the method for constructing water body comprehensive toxicity characterization parameters by algae photosynthesis inhibition method according to claim 1, wherein in the step 3, O-J, O-I section OJIP curve and F0 are used for forming area S of a closed areaJ、SIThe maximum area enclosed by the oji curve and F-F0, i.e. the enclosed area S of the O-P curvePIn which S isJ、SI、SPThe calculation method is shown in formulas (5), (6) and (7):
5. the method for constructing water body comprehensive toxicity characterization parameters by algae photosynthesis inhibition method according to claim 1, wherein in the step 4, the relative area of the enclosed area of the O-J, O-I section curve under the maximum enclosed area is further calculated, and S is utilizedPTo SJ、SICarrying out normalization processing to obtain two curve segment data items SJ *、SI *Extracting the ascending process information of O-J and J-I curves and eliminating the influence of the ascending process information on the absolute intensity of the curve measurement, wherein SJ *、SI *The calculation method is shown in formulas (8) and (9):
6. the method for constructing water body comprehensive toxicity characterization parameters through algae photosynthesis inhibition method according to claim 4, wherein in the step 5, characteristic point information F of J, I is further includedJ *、FI *And O-J and J-I section curve process information SJ *、SI *And fusion, wherein the fusion method adopts the mode that each characteristic point data item is multiplied by a curve section data item and then accumulated: sJ *×FJ *+SI *×FI *。
7. The method for constructing water body comprehensive toxicity characterization parameters through algae photosynthesis inhibition according to claim 5, wherein the step 5 further comprises SJ *×FJ *+SI *×FI *Based on the time information T of J, I characteristic pointsFJ、TFIFor J, I feature point data item FJ *And FI *Making correction by using O-J and J-I section curve process time period length TFJ-TF0、TFI-TF0For two curve segment data items SJ *And SI *Correcting to finally construct a water body comprehensive toxicity characterization parameter PICTE：
Wherein, the occurrence time sequence of different characteristic sites in OJIP is known, the corresponding time period lengths of O-J and J-I curves are known to be different, and the intensity of different characteristic sites and the response capability and sensitivity of curve rising process to toxic substances are different to a certain extent, so that each characteristic site data item F is usedJ *、FI *Divided by respective times of occurrence TFJ、TFITo J,I, correcting the feature point data item; two curve segment data items SJ *、SI *Divided by the respective corresponding time interval length TFJ-TF0、TFI-TF0The O-J and J-I curve segment data items are corrected.
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