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Screening method of high flux 96 orifice plate for herbicide

A herbicide, high-throughput technology, applied in biochemical equipment and methods, measuring devices, and microbial determination/inspection, etc., to achieve the effects of reducing usage, easy operation and high sensitivity

Inactive Publication Date: 2008-02-20
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For algae cells, due to their microbial characteristics, the probability of trait changes due to external factors is relatively high, so there are no related reports on the use of 96-well plate method and liquid medium to culture and analyze algae cells

Method used

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  • Screening method of high flux 96 orifice plate for herbicide
  • Screening method of high flux 96 orifice plate for herbicide
  • Screening method of high flux 96 orifice plate for herbicide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] In the exponential growth phase test of Chlorella vulgaris (Chlorella vulgaris, purchased from the Institute of Hydrobiology, Chinese Academy of Sciences, the same below) using Shuisheng No. 4 medium, Chlorella was inoculated in a 250 μL 96-well plate with a total volume of 200 μL. The initial inoculated number of Chlorella ranges from 7 to 8×10 5 individual / mL. After the inoculation, it was treated with a certain concentration gradient of the herbicide fluazifop-ethyl (purchased from Jiangsu Qiayi Agrochemical Co., Ltd.), and the concentration gradient was 0, 0.05, 0.1, 0.25, 0.5, 1, 5mg / L, and each 4 parallel samples were set for each concentration. Illumination culture conditions: temperature (25±0.2) ℃, light 2000Lx, continuous light, seal with plastic wrap, no nutrient solution was added during the test, the algae solution in the 96-well plate was inflated 4-5 times a day with a row gun, and co-cultivated for 72 hours , Test its absorbance at 630nm with a Bio-Rad...

Embodiment 2

[0030] In the exponential growth phase test of Chlorella vulgaris (Chlorella vulgaris, purchased from the Institute of Hydrobiology, Chinese Academy of Sciences) using Aquatic No. The number of algae ranges from 7 to 8×10 5individual / mL. After the inoculation, it was processed with a certain concentration gradient of the herbicide fluroxypyr (purchased from Zhejiang Jiema Chemical Co., Ltd.), the concentration gradient was 0, 0.05, 0.1, 0.25, 0.5, 1, 5mg / L, each concentration Set up 4 parallel samples. Illumination culture conditions: temperature (25±0.2) ℃, light 2000Lx, continuous light, seal with plastic wrap, no nutrient solution was added during the test, the algae solution in the 96-well plate was inflated 4-5 times a day with a row gun, and co-cultivated for 72 hours , test its absorbance at 630nm with a Bio-Rad 680 microplate reader every day, and calculate the EC of the herbicide on the algae according to the relationship between the absorbance and the algae biomass...

Embodiment 3

[0034] In the exponential growth phase test of Chlorella vulgaris (Chlorella vulgaris, purchased from the Institute of Hydrobiology, Chinese Academy of Sciences) using Aquatic No. The number of algae ranges from 7 to 8×10 5 individual / mL. After inoculation, it was treated with a certain concentration gradient of the herbicide acetochlor (purchased from Hangzhou Qingfeng Agrochemical Co., Ltd.). Parallel samples, while setting a blank sample control. Illumination culture conditions: temperature (25±0.2) ℃, light 2000Lx, continuous light, seal with plastic wrap, no nutrient solution was added during the test, the algae solution in the 96-well plate was inflated 4-5 times a day with a row gun, and co-cultivated for 72 hours , test its absorbance at 630nm with a Bio-Rad 680 microplate reader every day, and calculate the EC of the herbicide on the algae according to the relationship between the absorbance and the algae biomass. 50 .

[0035] At the same time, the EC of herbicid...

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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 [0001] The invention relates to a 96-well plate screening method for high throughput of herbicides. (2) Background technology [0002] Pesticides play an important role in agricultural and forestry production and sanitary pest control in my country and the world. However, with the increasing demand for environmental and consumer safety, and the emergence of pest resistance, the service life of pesticides is shortened, making the creation of new pesticides And development is becoming more and more urgent and difficult. According to statistics, the successful development of a high-efficiency, low-toxicity, and environment-friendly new pesticide currently requires the synthesis and screening of 80,000 compounds, costing hundreds of millions of dollars. This poses new challenges to the synthesis and screening of compounds in drug development. It is very difficult to screen a large number of compounds in a short period of time with conventional greenhouse bio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/17C12Q1/08
Inventor 张安平叶璟刘维屏严航贞蒋凌峰聂利宾
Owner ZHEJIANG UNIV OF TECH
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