CN101126712A - Screening method of high flux 96 orifice plate for herbicide - Google Patents
Screening method of high flux 96 orifice plate for herbicide Download PDFInfo
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- CN101126712A CN101126712A CNA200710071203XA CN200710071203A CN101126712A CN 101126712 A CN101126712 A CN 101126712A CN A200710071203X A CNA200710071203X A CN A200710071203XA CN 200710071203 A CN200710071203 A CN 200710071203A CN 101126712 A CN101126712 A CN 101126712A
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- 238000000034 method Methods 0.000 title claims abstract description 51
- 230000002363 herbicidal effect Effects 0.000 title claims abstract description 48
- 239000004009 herbicide Substances 0.000 title claims abstract description 46
- 238000012216 screening Methods 0.000 title claims abstract description 16
- 230000004907 flux Effects 0.000 title abstract 2
- 241000195649 Chlorella <Chlorellales> Species 0.000 claims abstract description 40
- 238000002835 absorbance Methods 0.000 claims abstract description 30
- 230000003203 everyday effect Effects 0.000 claims abstract description 25
- 238000012360 testing method Methods 0.000 claims abstract description 21
- 235000015097 nutrients Nutrition 0.000 claims abstract description 20
- 239000001963 growth medium Substances 0.000 claims abstract description 17
- 239000007788 liquid Substances 0.000 claims description 35
- 241000195493 Cryptophyta Species 0.000 claims description 33
- 238000011081 inoculation Methods 0.000 claims description 20
- 238000005286 illumination Methods 0.000 claims description 18
- 240000009108 Chlorella vulgaris Species 0.000 claims description 17
- 235000007089 Chlorella vulgaris Nutrition 0.000 claims description 17
- YUVKUEAFAVKILW-UHFFFAOYSA-N 2-(4-{[5-(trifluoromethyl)pyridin-2-yl]oxy}phenoxy)propanoic acid Chemical compound C1=CC(OC(C)C(O)=O)=CC=C1OC1=CC=C(C(F)(F)F)C=N1 YUVKUEAFAVKILW-UHFFFAOYSA-N 0.000 claims description 8
- 239000003755 preservative agent Substances 0.000 claims description 8
- 230000002335 preservative effect Effects 0.000 claims description 8
- 239000012496 blank sample Substances 0.000 claims description 4
- 239000013068 control sample Substances 0.000 abstract description 3
- 230000000694 effects Effects 0.000 abstract description 3
- 239000000523 sample Substances 0.000 abstract description 2
- 238000012258 culturing Methods 0.000 abstract 1
- 230000035945 sensitivity Effects 0.000 abstract 1
- 230000002401 inhibitory effect Effects 0.000 description 17
- 150000001875 compounds Chemical class 0.000 description 9
- 230000001186 cumulative effect Effects 0.000 description 7
- 239000005558 Fluroxypyr Substances 0.000 description 6
- MEFQWPUMEMWTJP-UHFFFAOYSA-N fluroxypyr Chemical compound NC1=C(Cl)C(F)=NC(OCC(O)=O)=C1Cl MEFQWPUMEMWTJP-UHFFFAOYSA-N 0.000 description 6
- VTNQPKFIQCLBDU-UHFFFAOYSA-N Acetochlor Chemical compound CCOCN(C(=O)CCl)C1=C(C)C=CC=C1CC VTNQPKFIQCLBDU-UHFFFAOYSA-N 0.000 description 5
- 239000002028 Biomass Substances 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 238000012417 linear regression Methods 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 239000000575 pesticide Substances 0.000 description 3
- 238000002798 spectrophotometry method Methods 0.000 description 3
- 244000207740 Lemna minor Species 0.000 description 2
- 235000006439 Lemna minor Nutrition 0.000 description 2
- 235000001855 Portulaca oleracea Nutrition 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000003905 agrochemical Substances 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000009509 drug development Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 238000007821 culture assay Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000009333 weeding Methods 0.000 description 1
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Abstract
The utility model provides a screening method used for herbicide high flux 96-hole orifice plate. The utility model adopts the technical proposal that: firstly, exponential phase chlorella is inoculated on the 96-hole orifice with a culture medium which is suitable for the chlorella, and the initial inoculating number of the chlorella is 7 to 8 x 105 per mL; secondly, the concentration gradient herbicide is dipped into each hole, and each concentration is provided with two to five duplicate samples as well as an empty control sample. The culturing condition is that: temperature is 25 plus or minus 0.2 DEG C, light is 2000Lx, continuous light is kept, fresh-keeping film is used to seal, no nutrient solution is added, chlorella solution is aerated four to five times every day, the chlorella is cultivated 72 to 144 hours in total, test sample is measured at 630nm absorbance every day, and then the EC50 (median effect concentration) of the herbicide on the chlorella is calculated according to the relationship between the absorbance and the herbicide concentration. Compared with the flask method, the utility model has the advantages of reducing herbicide amount in screening, and having easy operation, fast speed and high sensitivity.
Description
(1) technical field
The present invention relates to a kind of high-throughout 96 orifice plate screening techniques of herbicide that are used for.
(2) background technology
Agricultural chemicals is playing an important role aspect China and even global agriculture and forestry production and the sanitary insect pest control, but along with the cry of environment and consumption safety improves day by day, and the drug-fast generation of harmful organism makes agricultural chemicals shortening in serviceable life, make that the initiative of novel pesticide is more and more urgent, also more and more difficult with exploitation.According to statistics, the successful exploitation of efficient, low toxicity, environmentally friendly novel pesticide needs 80000 compounds of synthetic screening at present, costs more than one hundred million dollars.This synthetic and screening to compound in the drug development has all proposed new challenge.It is very difficult screening a large amount of compounds at short notice with the living survey method in the greenhouse of routine, and miniature screening method is not only tested scale and dwindled greatly, and pesticide dosage is few, breakneck acceleration is also very fast.With representative plant test, as algae, duckweed etc.The compound of doses and algae or duckweed are added in the triangular flask mixing respectively.Shaken cultivation under controlled conditions.Measure the optical density value of mixed liquor before and after cultivating, if optical density value is constant even minimizing, show that algae does not grow, compound has activity of weeding.Also can obtain EC50, to judge the compound activity size by growth inhibition ratio.Can also cultivate alga cells, the inhibiting effect of alga cells be judged its active size by compound.Along with further developing of science and technology, synthetic and screening to compound in the drug development is then had higher requirement, in order further to reduce cost, at the initial compound that is used to screen of obtaining in synthetic usually all at the order of magnitude of microgram, therefore need a kind of more miniature screening technique, to substitute existing triangular flask method.
At present existing 96 well plate method are applied to the report of herbicidal activity screening, adopt solid medium, implant weed seed, by measuring weeds fresh weight and plant height, come herbicidal activity is analyzed.And, because its microbiologic properties, influenced by extraneous factor and that the probability that proterties changes takes place is bigger for alga cells, therefore yet there are no utilization 96 well plate method, adopt fluid nutrient medium alga cells to be carried out the relevant report of culture assays.
(3) summary of the invention
The present invention then is for a kind of high-throughout 96 orifice plate screening techniques of herbicide that are used for are provided, to substitute traditional triangular flask method.
For reaching goal of the invention the technical solution used in the present invention be:
A kind of high-throughout 96 orifice plate screening techniques of herbicide that are used for, described method is as follows: adopt the nutrient culture media be applicable to chlorella, the chlorella of exponential phase is inoculated in 96 orifice plates, initial inoculation chlorella number scope is 7～8 * 10
5Individual/mL, the inoculation back adds the herbicide with concentration gradient toward each hole, and each concentration is provided with 2～5 parallel samples, and blank sample contrast is set simultaneously; Condition of culture: 25 ± 0.2 ℃ of temperature, illumination 2000Lx, continuous light, seal with preservative film, Ensure Liquid liquid not, every day is to algae liquid inflation 4～5 times, cultivate 72～144h altogether, every day, specimen was in the absorbance at 630nm place, according to the relation between absorbance and herbicide concentration, calculated the EC of herbicide to chlorella
50Find in the experiment that if algae liquid is not inflated in the incubation, then frustule easily is deposited on the hole wall, influences the accuracy of absorbance measuring; And, then can obviously improve this problem by the processing of every day to algae liquid inflation 4～5 times, the frustule growth conditions is good, is evenly distributed, and is difficult for precipitation.
Data processing, analytical approach: shaken cultivation under these conditions, with the nutrient solution is reference, measure light absorption value with blood counting chamber in the microscopically direct census and at maximum absorption wavelength 630nm, 3 repetitions are established in test, set up the linear regression relation of frustule concentration and absorbance.Inhibiting rate directly adopts (control sample absorbance-processing sample absorbance)/control sample absorbance to calculate, and sets up the linear regression relation of the natural logarithm (lnC) of inhibiting rate (P) and concentration, and to find the solution inhibiting rate be 50% concentration value (EC
50).
Described chlorella is preferably chlorella vulgaris (Chlorella vulgaris).
The described nutrient culture media that is applicable to chlorella is the common nutrient culture media that chlorella is cultivated that can be used for, and as aquatic No. four nutrient culture media, HA-SK nutrient culture media etc., is preferably aquatic No. four nutrient culture media.Described aquatic No. four nutrient culture media can use commercial product, also can prepare by following composition: (NH
4)
2SO
4, 0.20g; Ca
3(PO
4)
2, 0.03g; MgSO
47H
2O, 0.08g; NaHCO
3, 0.10g; KCl, 0.025g; 1% (mass concentration) FeCl
3, 0.15mL; Soil extract, 0.50mL; Water complements to 1000mL.
Concrete, described method is as follows: adopt aquatic No. four nutrient culture media, the chlorella vulgaris that will be in exponential phase is inoculated in 96 orifice plates, and the chlorella vulgaris initial concentration is 7～8 * 10
5Individual/m, the inoculation back adds efficient fluazifop, concentration gradient is 0,0.05,0.1,0.25,0.5,1,5mg/L, each concentration is provided with 4 parallel samples, condition of culture: 25 ± 0.2 ℃ of temperature, illumination 2000Lx, continuous light seals with preservative film, not Ensure Liquid liquid, inflate 4～5 time algae liquid in 96 orifice plates with the volley of rifle fire every day, cultivate 72h altogether, test it in 630nm place absorbance, according to relation between absorbance and algae biomass with Bio-Rad 680 type microplate reader every day, calculate the EC50 of herbicide to algae, the result gets its mean value.
Beneficial effect of the present invention is mainly reflected in: greatly reduced the herbicide use amount in the screening process, and easy and simple to handle, quick, highly sensitive.
(4) description of drawings
Fig. 1 is 96 well plate method inhibiting rates and Ln (C) matched curve (efficient fluazifop); When P=50%, C=1.35mg/L (P is an inhibiting rate, and C is a herbicide concentration, down together);
Fig. 2 is triangular flask method inhibiting rate and Ln (C) matched curve (efficient fluazifop); When P=50%, C=1.58mg/L;
Fig. 3 is 96 well plate method inhibiting rates and Ln (C) matched curve (fluroxypyr); When P=50%, C=0.60mg/L;
Fig. 4 is triangular flask method inhibiting rate and Ln (C) matched curve (fluroxypyr); When P=50%, C=0.66mg/L;
Fig. 5 is 96 well plate method inhibiting rates and Ln (C) matched curve (Acetochlor); When P=50%, C=0.33mg/L;
Fig. 6 is triangular flask method inhibiting rate and Ln (C) matched curve (Acetochlor); When P=50%, C=0.45mg/L;
Fig. 7 is 96 well plate method inhibiting rates and Ln (C) matched curve (Quizalotop-ethyl); When P=50%, C=2.10mg/L;
Fig. 8 is triangular flask method inhibiting rate and Ln (C) matched curve (Quizalotop-ethyl); When P=50%, C=1.91mg/L.
(5) embodiment
The present invention is described further below in conjunction with specific embodiment, but protection scope of the present invention is not limited in this:
Embodiment 1:
Adopt aquatic No. four nutrient culture media, (Chlorella vulgaris is available from Inst. of Hydrobiology, Chinese Academy of Sciences at chlorella vulgaris, test exponential phase down together), chlorella is inoculated in 250 μ L, 96 orifice plates, and cumulative volume is 200 μ L, and initial inoculation chlorella number scope is 7～8 * 10
5Individual/mL.Inoculation back is to its herbicide efficient fluazifop (available from being in harmony benefit farmingization company limited in Jiangsu) processing of carrying out the finite concentration gradient, and concentration gradient is 0,0.05,0.1,0.25,0.5,1,5mg/L, and each concentration is provided with 4 parallel samples.Illumination cultivation condition: temperature (25 ± 0.2) ℃, illumination 2000Lx, continuous light seals with preservative film, and duration of test is Ensure Liquid liquid not, inflate 4-5 time algae liquid in 96 orifice plates with the volley of rifle fire every day, cultivate 72h altogether, test it in 630nm place absorbance, according to relation between absorbance and algae biomass with Bio-Rad 680 type microplate reader every day, calculate the EC50 of herbicide to algae, the result gets its mean value.
The triangular flask method of the standard of recommending with ISO is measured the EC of herbicide to algae simultaneously
50, it is as follows that the triangular flask method is measured process: chlorella vulgaris is inoculated in the triangular flask, and cumulative volume is 75mL, and initial inoculation chlorella number scope is 7～8 * 10
5Individual/mL.Handle the herbicide efficient fluazifop that it carries out the finite concentration gradient inoculation back, and concentration gradient is 0,0.05,0.1,0.25,0.5,1,5mg/L, and each concentration is provided with 4 parallel samples, and the result gets its mean value.Illumination cultivation condition: temperature (25 ± 0.2) ℃, illumination 2000Lx, continuous light, seal with gauze, duration of test is Ensure Liquid liquid not, and manually vibrate 4-5 time every day, cultivate 72h altogether, according to the relation between absorbance and herbicide concentration, calculate the EC of herbicide to algae with the absorbance of spectrophotometric determination algae liquid at the 690nm place every day
50
Shaken cultivation is a reference with the nutrient solution under these conditions, measures light absorption value with blood counting chamber in the microscopically direct census and at maximum absorption wavelength 630nm, and 3 repetitions are established in test, set up the linear regression relation (seeing Table 1) of frustule concentration and absorbance:
Table 1: the linear regression relation of cell concentration and absorbance
| Assay method | Linear equation | R 2 | P |
| The triangular flask method | Y＝109.56X+5.66 | 0.9872 | 0.0017 |
| 96 well plate method | Y＝78.38X+7.78 | 0.9627 | 0.0039 |
Inhibiting rate (P) is seen Fig. 1, Fig. 2 with the matched curve of the natural logarithm (Ln (C)) of herbicide concentration.
Measurement result: the efficient fluazifop that calculates with 96 well plate method is to the EC of chlorella
50Be 1.35mg/L, the efficient fluazifop that calculates with the triangular flask method is to the EC of chlorella
50Be 1.58mg/L.
Embodiment 2:
Adopt aquatic No. four nutrient culture media, in the test exponential phase of chlorella vulgaris (Chlorella vulgaris is available from Inst. of Hydrobiology, Chinese Academy of Sciences), chlorella is inoculated in the 250 μ L96 orifice plates, cumulative volume is 200 μ L, and initial inoculation chlorella number scope is 7～8 * 10
5Individual/mL.Handle the herbicide fluroxypyr (available from the prompt horse chemical industry in Zhejiang company limited) that it carries out the finite concentration gradient inoculation back, and concentration gradient is 0,0.05,0.1,0.25,0.5,1,5mg/L, and each concentration is provided with 4 parallel samples.Illumination cultivation condition: temperature (25 ± 0.2) ℃, illumination 2000Lx, continuous light, seal with preservative film, duration of test is Ensure Liquid liquid not, and inflate 4-5 time algae liquid in 96 orifice plates with the volley of rifle fire every day, cultivate 72h altogether, test it in 630nm place absorbance with Bio-Rad 680 type microplate reader every day, according to the relation between absorbance and algae biomass, calculates the EC of herbicide to algae
50
Triangular flask method step is with embodiment 1.Inhibiting rate (P) is seen Fig. 3, Fig. 4 with the matched curve of the natural logarithm (Ln (C)) of herbicide concentration.
Measurement result: the fluroxypyr that calculates with 96 well plate method is to the EC of chlorella
50Be 0.60mg/L, the fluroxypyr that calculates with the triangular flask method is to the EC of chlorella
50Be 0.66mg/L.
Embodiment 3:
Adopt aquatic No. four nutrient culture media, in the test exponential phase of chlorella vulgaris (Chlorella vulgaris is available from Inst. of Hydrobiology, Chinese Academy of Sciences), chlorella is inoculated in the 250 μ L96 orifice plates, cumulative volume is 200 μ L, and initial inoculation chlorella number scope is 7～8 * 10
5Individual/mL.Handle the herbicide Acetochlor (available from Qingfeng Agrochemical Co., Ltd) that it carries out the finite concentration gradient inoculation back, and concentration gradient is 0,0.05,0.1,0.25,0.5,1mg/L, and each concentration is provided with 4 parallel samples, and blank sample contrast is set simultaneously.Illumination cultivation condition: temperature (25 ± 0.2) ℃, illumination 2000Lx, continuous light, seal with preservative film, duration of test is Ensure Liquid liquid not, and inflate 4-5 time algae liquid in 96 orifice plates with the volley of rifle fire every day, cultivate 72h altogether, test it in 630nm place absorbance with Bio-Rad 680 type microplate reader every day, according to the relation between absorbance and algae biomass, calculates the EC of herbicide to algae
50
The triangular flask method of the standard of recommending with ISO is measured the EC of herbicide to algae simultaneously
50, the triangular flask method is measured the following chlorella of process and is inoculated in the triangular flask, and cumulative volume is 75mL, and initial inoculation chlorella number scope is 7～8 * 10
5Individual/mL.Handle the herbicide Acetochlor that it carries out the finite concentration gradient inoculation back, and concentration gradient is 0,0.05,0.1,0.25,0.5,1,5mg/L, and each concentration is provided with 4 parallel samples, and the result gets its mean value.Illumination cultivation condition: temperature (25 ± 0.2) ℃, illumination 2000Lx, continuous light, seal with gauze, duration of test is Ensure Liquid liquid not, and manually vibrate 4-5 time every day, cultivate 72h altogether, according to the relation between absorbance and herbicide concentration, calculate the EC of herbicide to algae with the absorbance of spectrophotometric determination algae liquid at the 690nm place every day
50
Inhibiting rate (P) is seen Fig. 5, Fig. 6 with the matched curve of the natural logarithm (Ln (C)) of herbicide concentration.
Measurement result: the Acetochlor that calculates with 96 well plate method is to the EC of chlorella
50Be 0.33mg/L, the fluroxypyr that calculates with the triangular flask method is to the EC of chlorella
50Be 0.45mg/L.
Embodiment 4:
Adopt aquatic No. four nutrient culture media, in the test exponential phase of chlorella vulgaris (Chlorella vulgaris is available from Inst. of Hydrobiology, Chinese Academy of Sciences), chlorella is inoculated in 250 μ L, 96 orifice plates, cumulative volume is 200 μ L, and initial inoculation chlorella number scope is 7～8 * 10
5Individual/mL.Handle the herbicide Quizalotop-ethyl (available from Jiangsu Fengshan Group Co) that it carries out the finite concentration gradient inoculation back, and concentration gradient is 0,0.1,0.25,0.5,1,5mg/L, and each concentration is provided with 4 parallel samples, and blank sample contrast is set simultaneously.Illumination cultivation condition: temperature (25 ± 0.2) ℃, illumination 2000Lx, continuous light, seal with preservative film, duration of test is Ensure Liquid liquid not, and inflate 4-5 time algae liquid in 96 orifice plates with the volley of rifle fire every day, cultivate 72h altogether, test it in 630nm place absorbance with Bio-Rad 680 type microplate reader every day, according to the relation between absorbance and algae biomass, calculates the EC of herbicide to algae
50
The triangular flask method of the standard of recommending with ISO is measured the EC of herbicide to algae simultaneously
50, the triangular flask method is measured the following chlorella of process and is inoculated in the triangular flask, and cumulative volume is 75mL, and initial inoculation chlorella number scope is 7～8 * 10
5Individual/mL.Handle the herbicide Quizalotop-ethyl that it carries out the finite concentration gradient inoculation back, and concentration gradient is 0,0.05,0.1,0.25,0.5,1,5mg/L, and each concentration is provided with 4 parallel samples, and the result gets its mean value.Illumination cultivation condition: temperature (25 ± 0.2) ℃, illumination 2000Lx, continuous light, seal with gauze, duration of test is Ensure Liquid liquid not, and manually vibrate 4-5 time every day, cultivate 72h altogether, according to the relation between absorbance and herbicide concentration, calculate the EC of herbicide to algae with the absorbance of spectrophotometric determination algae liquid at the 690nm place every day
50
Inhibiting rate (P) is seen Fig. 7, Fig. 8 with the matched curve of the natural logarithm (Ln (C)) of herbicide concentration.
Measurement result: the Quizalotop-ethyl that calculates with 96 well plate method is to the EC of chlorella
50Be 2.10mg/L, the Quizalotop-ethyl that calculates with the triangular flask method is to the EC of chlorella
50Be 1.91mg/L.
Claims (4)
1. one kind is used for the high-throughout 96 orifice plate screening techniques of herbicide, and described method is as follows: adopt the nutrient culture media be applicable to chlorella, the chlorella of exponential phase is inoculated in 96 orifice plates, initial inoculation chlorella number scope is 7～8 * 10
5Individual/mL, the inoculation back adds the herbicide with concentration gradient toward each hole, and each concentration is provided with 2～5 parallel samples, and blank sample contrast is set simultaneously; Condition of culture: 25 ± 0.2 ℃ of temperature, illumination 2000Lx, continuous light, seal with preservative film, Ensure Liquid liquid not, every day is to algae liquid inflation 4～5 times, cultivate 72～144h altogether, every day, specimen was in the absorbance at 630nm place, according to the relation between absorbance and herbicide concentration, calculated the EC of herbicide to chlorella
50
2. the method for claim 1 is characterized in that described chlorella is chlorella vulgaris (Chlorella vulgaris).
3. the method for claim 1 is characterized in that describedly being applicable to that the nutrient culture media of chlorella is aquatic No. four nutrient culture media.
4. the method for claim 1, it is characterized in that described method is as follows: adopt aquatic No. four nutrient culture media, the chlorella vulgaris that will be in exponential phase is inoculated in 96 orifice plates, and the chlorella vulgaris initial concentration is 7～8 * 10
5Individual/m, the inoculation back adds efficient fluazifop, concentration gradient is 0,0.05,0.1,0.25,0.5,1,5mg/L, each concentration is provided with 4 parallel samples, condition of culture: 25 ± 0.2 ℃ of temperature, illumination 2000Lx, continuous light seals with preservative film, not Ensure Liquid liquid, inflate 4～5 time algae liquid in 96 orifice plates with the volley of rifle fire every day, cultivate 72h altogether, test it in 630nm place absorbance with Bio-Rad 680 type microplate reader every day, according to the relation between absorbance and herbicide concentration, calculate the EC50 of herbicide to algae, the result gets its mean value.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103074205A (en) * | 2011-10-26 | 2013-05-01 | 中国海洋石油总公司 | Electric turntable system, microalgae high-throughput screening device, and microalgae high-throughput screening method |
| CN104990878A (en) * | 2015-07-02 | 2015-10-21 | 上海应用技术学院 | Cell inhibition ratio measuring method for green microcystis |
| CN109722388A (en) * | 2019-03-13 | 2019-05-07 | 厦门大学 | Microalgae commensalism bacterium isolation medium, separation method and the crucial bacterium high-throughput screening method for influencing micro algae growth |
| WO2021090182A1 (en) * | 2019-11-04 | 2021-05-14 | Oxford University Innovation Limited | Identification and characterisation of herbicides and plant growth regulators |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU774776B2 (en) * | 1999-04-15 | 2004-07-08 | University Of Virginia Patent Foundation | Proteome mining |
| CN1234277C (en) * | 2003-03-19 | 2006-01-04 | 浙江大学 | Method for preparing high efficiency biological weed control bacterial agent and its usage |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103074205A (en) * | 2011-10-26 | 2013-05-01 | 中国海洋石油总公司 | Electric turntable system, microalgae high-throughput screening device, and microalgae high-throughput screening method |
| CN104990878A (en) * | 2015-07-02 | 2015-10-21 | 上海应用技术学院 | Cell inhibition ratio measuring method for green microcystis |
| CN109722388A (en) * | 2019-03-13 | 2019-05-07 | 厦门大学 | Microalgae commensalism bacterium isolation medium, separation method and the crucial bacterium high-throughput screening method for influencing micro algae growth |
| WO2021090182A1 (en) * | 2019-11-04 | 2021-05-14 | Oxford University Innovation Limited | Identification and characterisation of herbicides and plant growth regulators |
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