CN102094063A - Method for detecting water quality genetic toxicity by broad bean root tip micronucleus test - Google Patents
Method for detecting water quality genetic toxicity by broad bean root tip micronucleus test Download PDFInfo
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- CN102094063A CN102094063A CN 201010580785 CN201010580785A CN102094063A CN 102094063 A CN102094063 A CN 102094063A CN 201010580785 CN201010580785 CN 201010580785 CN 201010580785 A CN201010580785 A CN 201010580785A CN 102094063 A CN102094063 A CN 102094063A
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Abstract
The invention discloses a method for detecting water quality genetic toxicity by a broad bean root tip micronucleus test, comprising the following steps of: soaking seeds for Songrun green-peel beans, accelerating germination, and obtaining a seed of which the primary root has the length of 1.5-2cm; respectively processing seed root tips by Cr6+ solutions of different concentrations; repairing, fixing and dyeing root tip cells; carrying out microscopic examination to the root tips, and calculating the permillage of cell micronucleus processed by the Cr6+ solutions of different concentrations; drawing the standard curve for the permillage of cell micronucleus relative to Cr6+ concentration; then, measuring the permillage of cell micronucleus of a water sample to be detected with the same method; and substituting the standard curve to obtain the water quality toxicity of the water sample to be detected, which is represented by Cr6+ concentration. In the method, Cr6+ is selected as the standard toxic substance, and the quantitative evaluation of the water quality toxicity is realized by the standard curve of the micronucleus permillage relative to Cr+6. The genetic toxicity of the water quality can be more intuitively and accurately detected by the method, results are easy to unify, and the repeatability is good.
Description
Technical field
The present invention relates to a kind of detection method of water quality toxicity, be specifically related to a kind of broad bean tip of a root micronucleus test and detect the genotoxic method of water quality.
Background technology
In order to ensure safe drinking water, reduce the threat that environmental degradation brings to drinking-water quality, people often adopt the whole bag of tricks that water quality is estimated, the health risk that causes with effective grasp tap water.
Micronucleus test is one of important method that detects chromosome aberration, since nineteen eighty-two is set up by Degressi etc., advantages such as, reliable results easy, quick with it are widely used in the mutagenicity detection to chemical toxic substance and environmental pollutant in the environment such as water source, soil, atmosphere.Just list broad bean tip of a root micronucleus test detection technique in " environment measuring technical specifications " as far back as State Bueau of Environmental Protection in 1986 and be used for the water surrounding detection technique.Studies show that micronucleus test result and Salmonella reversion test have certain dependency, and the micronuclear rates that is brought out exists dose-effect relationship within the specific limits, be suitable for the quantitative evaluation water quality toxicity.The shortcoming of existing evaluation technique: but most of micronucleus test just adopts pollution index quantitative evaluation water quality toxicity, toxic reaction is directly perceived inadequately, and toxic grade is divided certain subjectivity.
Summary of the invention
The present invention is directed to the deficiency of existing water quality genetoxic detection method, provide a kind of broad bean tip of a root micronucleus test to detect the genotoxic method of water quality, this method is with Cr
6+As the standard toxicant, when the evaluating water quality genetoxic with Cr
6+Concentration represent the genetoxic of water quality, realized the quantification of water quality toxicity, and detected result is directly perceived more, clear and definite, be easy to unified, for the water quality toxicity quantitative evaluation provides strong support.
The present invention is achieved by the following measures:
The present invention selects Cr
6+As the standard toxicant, hexavalent chromium is a kind of in World Health Organization's carcinogens list, belongs to the toxicity index in drinking water sanitary standard, and limit value is 0.05mg/L, and the limit value of different grades is also arranged in water environment quality standard, therefore uses Cr
6+It is unified that concentration represents that the water quality toxicity result is easy to, it is operating as: at first, select the chromium ion concentration scope, selection principle is: the micronucleus quantity that can cause different quantities, with the chromium ion solution of different concns the broad bean tip of a root is handled then, obtained the typical curve of chromium ion by micronucleus test again the bean root tip cell microkernel rate.Use the same method then water sample to be measured is carried out broad bean tip of a root micronucleus test, the micronuclear rates substitution typical curve of gained can be obtained the water sample toxicity represented with chromium ion concentration, this method has been avoided the influence that other empirical factors produced, make water quality toxicity more accurate, detected result is easier to unified.
In order to solve the problems of the technologies described above, the present invention adopts following steps:
A kind of broad bean tip of a root micronucleus test detects the genotoxic method of water quality, it is characterized in that may further comprise the steps at least:
(1) be indicator organism with Songzi rascal beans, by seed soaking, vernalization, getting primary root is the long Songzi rascal beans seed of 1.5cm～2cm;
(2) Cr of usefulness different concns
6+Solution is handled the tip of a root of Songzi rascal beans seed respectively, behind root-tip cells reparation, fixing, staining procedure, the tip of a root is carried out microscopy then, calculates different concns Cr
6+The cell micronucleus permillage of solution-treated;
(3) with Cr
6+Concentration is X-coordinate, and the cell micronucleus permillage is an ordinate zou, draws Cr
6+The typical curve of concentration pair cell micronucleus permillage;
(4) adopt the Songzi rascal beans that use the same method to handling through water sample to be measured and carry out micronucleus test, calculate the cell micronucleus permillage, the cell micronucleus permillage substitution typical curve with water sample gained to be measured can obtain Cr
6+The water quality toxicity of the water sample to be measured that concentration is represented.
In the aforesaid method, the concrete operations of step (2) are: with the Cr of different concns
6+Solution is handled 5-7h to the tip of a root of Songzi rascal beans seed respectively, after the processing Songzi rascal beans seed is embathed totally with deionized water respectively, root-tip cells is repaired 22h～24h, downcut the young root of 1cm then from tip of a root top, add fixedly 24-48h of Ka Nuoshi stationary liquid, with the Feulgen procedure 5-6h that dyes, under biomicroscope, root-tip cells is carried out microscopy at last, calculate different concns Cr
6+The cell micronucleus permillage of solution-treated.
In the above-mentioned steps (2), preferred, the tip of a root is handled 6h, root-tip cells is repaired 24h, the tip of a root adds Ka Nuoshi stationary liquid fixedly 24h, dyeing 5h.
In the above-mentioned steps (1), seed soaking time is 24h～36h, and germination time is 48-72h.
In the above-mentioned steps (1), preferred, seed soaking time is 24h, and germination time is 60h, and the primary root of Songzi rascal beans seed is 1.5cm.
The invention has the beneficial effects as follows: select Cr
6+For the standard toxicant, pass through Cr
6+The typical curve of micronuclear rates is realized quantitative evaluation to water quality toxicity, method of the present invention can more intuitive and accurate detection water quality genetoxic, the result is easy to unified, favorable reproducibility, can make judgement to the water pollution situation, for the water technology of sudden water pollution and water factory provides support, reduce because tap water exposes the human carcinogenic risk that causes.
Description of drawings
Fig. 1 is among the embodiment 1Cr
6+The typical curve of concentration pair cell micronucleus permillage.
Embodiment
Below by specific embodiment the present invention is further set forth, should be understood that, following explanation only is in order to explain the present invention, its content not to be limited.
The used chromium ion solution of the present invention all is to adopt potassium bichromate formulated.
Embodiment 1
This specific embodiment adopts following steps:
(1) select Songzi rascal beans as indicator organism;
(2) by seed soaking 24h, vernalization 60h is after the primary root of seed grows to 1.5cm, at Cr
6+Get 5 concentration point in the scope of concentration 0-0.015mg/L and respectively the tip of a root is handled 6h, the Cr of these 5 concentration point
6+Concentration is respectively 0mg/L, 0.025mg/L, 0.05mg/L, 0.1mg/L, 0.15mg/L; Seeds treated is embathed clean back with deionized water root-tip cells is repaired 24h, downcut the young root of 1cm afterwards from tip of a root top, add fixedly 24h of Ka Nuoshi stationary liquid, after behind the Feulgen procedure dyeing 5h, under biomicroscope, root-tip cells is carried out microscopy, Cr
6+The concentration cell micronucleus permillage of gained from small to large is respectively 0.25 ‰, 4.25 ‰, 7.75 ‰, 10.33 ‰, 12.67 ‰;
(3) with Cr
6+Concentration is X-coordinate, and the micronucleus permillage is an ordinate zou, draws Cr
6+The typical curve of concentration pair cell micronucleus permillage, as shown in Figure 1;
(4) get daily city tap-water and carry out broad bean tip of a root experiment, the cell micronucleus permillage that the gained water sample produces is 2.33 ‰, this water sample to the toxic action sizableness of broad bean in the Cr of 0.014mg/L
6+Action effect.
This specific embodiment adopts following steps:
(1) select Songzi rascal beans as indicator organism;
(2) by seed soaking 36h, vernalization 48h is after the primary root of seed grows to 2cm, at Cr
6+Get 5 concentration point in the scope of concentration 0-0.015mg/L and respectively the tip of a root is handled 5h, the Cr of these 5 concentration point
6+Concentration is respectively 0mg/L, 0.025mg/L, 0.05mg/L, 0.1mg/L, 0.15mg/L; Seeds treated is embathed clean back with deionized water root-tip cells is repaired 22h, downcut the young root of 1cm afterwards from tip of a root top, add fixedly 48h of Ka Nuoshi stationary liquid, after behind the Feulgen procedure dyeing 6h, under biomicroscope, root-tip cells is carried out microscopy, Cr
6+The concentration cell micronucleus permillage of gained from small to large is respectively 0.25 ‰, 4.26 ‰, 7.78 ‰, 10.38 ‰, 12.70 ‰;
(3) with Cr
6+Concentration is X-coordinate, and the micronucleus permillage is an ordinate zou, draws Cr
6+The typical curve of concentration pair cell micronucleus permillage;
(4) the factory's water that advances of getting daily city water factory carries out the experiment of the broad bean tip of a root, and the cell micronucleus permillage that the gained water sample produces is 3 ‰, this water sample to the toxic action sizableness of broad bean in the Cr of 0.015mg/L
6+Action effect.
Embodiment 3
This specific embodiment adopts following steps:
(1) select Songzi rascal beans as indicator organism;
(2) by seed soaking 24h, vernalization 50-72h is after the primary root of seed grows to 1.5cm, at Cr
6+Get 5 concentration point in the scope of concentration 0-0.015mg/L and respectively the tip of a root is handled 6h, the Cr of these 5 concentration point
6+Concentration is respectively 0mg/L, 0.025mg/L, 0.05mg/L, 0.1mg/L, 0.15mg/L; Seeds treated is embathed clean back with deionized water root-tip cells is repaired 24h, downcut the young root of 1cm afterwards from tip of a root top, add fixedly 24h of Ka Nuoshi stationary liquid, after behind the Feulgen procedure dyeing 5h, under biomicroscope, root-tip cells is carried out microscopy, Cr
6+The concentration cell micronucleus permillage of gained from small to large is respectively 0.25 ‰, 4.26 ‰, 7.77 ‰, 10.35 ‰, 12.69 ‰;
(3) with Cr
6+Concentration is X-coordinate, and the micronucleus permillage is an ordinate zou, draws Cr
6+The typical curve of concentration pair cell micronucleus permillage;
(4) the technology water outlet of getting daily city water factory is carried out the experiment of the broad bean tip of a root, and the cell micronucleus permillage that the gained water sample produces is 3.75 ‰, this water sample to the toxic action sizableness of broad bean in the Cr of 0.022mg/L
6+Action effect.
Claims (5)
1. a broad bean tip of a root micronucleus test detects the genotoxic method of water quality, it is characterized in that may further comprise the steps at least:
(1) be indicator organism with Songzi rascal beans, by seed soaking, vernalization, getting primary root is the long Songzi rascal beans seed of 1.5cm～2cm;
(2) Cr of usefulness different concns
6+Solution is handled the tip of a root of Songzi rascal beans seed respectively, behind root-tip cells reparation, fixing, staining procedure, the tip of a root is carried out microscopy then, calculates different concns Cr
6+The cell micronucleus permillage of solution-treated;
(3) with Cr
6+Concentration is X-coordinate, and the cell micronucleus permillage is an ordinate zou, draws Cr
6+The typical curve of concentration pair cell micronucleus permillage;
(4) adopt the Songzi rascal beans that use the same method to handling through water sample to be measured and carry out micronucleus test, calculate the cell micronucleus permillage, the cell micronucleus permillage substitution typical curve with water sample gained to be measured can obtain Cr
6+The water quality toxicity of the water sample to be measured that concentration is represented.
2. method according to claim 1 is characterized in that the concrete operations of step (2) are: with the Cr of different concns
6+Solution is handled 5-7h to the tip of a root of Songzi rascal beans seed respectively, after the processing Songzi rascal beans seed is embathed totally with deionized water respectively, root-tip cells is repaired 22h～24h, downcut the young root of 1cm then from tip of a root top, add fixedly 24-48h of Ka Nuoshi stationary liquid, with the Feulgen procedure 5-6h that dyes, under biomicroscope, root-tip cells is carried out microscopy at last, calculate different concns Cr
6+The cell micronucleus permillage of solution-treated.
3. method according to claim 2 is characterized in that: in the step (2), the tip of a root is handled 6h, root-tip cells is repaired 24h, the tip of a root adds Ka Nuoshi stationary liquid fixedly 24h, dyeing 5h.
4. according to each described method among the claim 1-3, it is characterized in that: in the step (1), seed soaking time is 24h～36h, and germination time is 48-72h.
5. method according to claim 4 is characterized in that: in the step (1), seed soaking time is 24h, and germination time is 60h, and the primary root of Songzi rascal beans seed is 1.5cm.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103018419A (en) * | 2012-12-18 | 2013-04-03 | 南京大学 | Method for determining risk of toxic pollutants in river water of high content of sand |
| CN104198678A (en) * | 2014-09-17 | 2014-12-10 | 山东农业大学 | Set of methods suitable for evaluating toxicity changes of pesticide residues in soil |
| CN105181936A (en) * | 2015-09-25 | 2015-12-23 | 南京大学 | Method for evaluating soil heavy metal pollution degree |
-
2010
- 2010-12-09 CN CN 201010580785 patent/CN102094063A/en active Pending
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| 《环境与健康杂志》 20040331 韩建英等 含铬工业废水对地下水及饮用水致突变性的影响 全文 1-5 第21卷, 第2期 * |
| 《环境与健康杂志》 20090731 吴丽娜等 五种重金属的蚕豆根尖微核试验及污染评价 618-620 1-5 第26卷, 第7期 * |
| 《环境科学与管理》 20081130 何芬 水质毒性检测方法的对比研究 全文 1-5 第33卷, 第11期 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103018419A (en) * | 2012-12-18 | 2013-04-03 | 南京大学 | Method for determining risk of toxic pollutants in river water of high content of sand |
| CN103018419B (en) * | 2012-12-18 | 2015-04-08 | 南京大学 | Method for determining risk of toxic pollutants in river water of high content of sand |
| CN104198678A (en) * | 2014-09-17 | 2014-12-10 | 山东农业大学 | Set of methods suitable for evaluating toxicity changes of pesticide residues in soil |
| CN105181936A (en) * | 2015-09-25 | 2015-12-23 | 南京大学 | Method for evaluating soil heavy metal pollution degree |
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Application publication date: 20110615 |