NY/T 1156.1-2006 Guidelines for Indoor Bioassay Tests for Pesticides Fungicides Part 1: Test for Inhibition of Germination of Pathogenic Fungal Spores Concave Slide Method NY/T1156.1-2006 Standard Download Decompression Password: www.bzxz.net

ICS65.100
Agricultural Industry Standard of the People's Republic of China
NY/T1156.1—2006
Guidelines for Laboratory Bioassay Tests of Pesticides
Fungicides
Part 1: Test for Inhibition of Pathogenic Fungal Spore Germination
Concave Slides
Pesticides guidelines for laboratory bioactivity testsPart 1: Determining fungicide inhibition of pathogen spore germinationonconcaweslides
Published on July 10, 2006
Implemented on October 1, 2006
Published by the Ministry of Agriculture of the People's Republic of China
"Guidelines for Laboratory Bioassay Tests of Pesticides Fungicides" is a series of standards. This part is the first part of "Guidelines for Laboratory Bioassay Tests of Pesticides Fungicides". This standard is proposed and managed by the Ministry of Agriculture of the People's Republic of China. The drafting unit of this standard: Pesticide Control Institute of the Ministry of Agriculture. The main drafters of this standard are: Xu Wenping, Zhu Chunyu, Wu Xinping, Zhang Hong, Qiu Jianping. The Ministry of Agriculture's Pesticide Testing Institute is responsible for the interpretation of this standard. NY/T1156.1—2006
1 Scope
Guidelines for Indoor Bioassay Tests for Pesticides Fungicides NY/T1156.1—2006
Part 1: Test for Inhibition of Germination of Pathogenic Fungal Spores Concave Slide Method This part specifies the basic requirements and methods for the test of fungicides for inhibition of germination of pathogenic fungal spores. This part is applicable to the indoor test for pesticide registration for determination of the inhibitory effect of fungicides on the germination of pathogenic fungal spores. Other tests shall be carried out in accordance with this part.
2 Test conditions
2.1 Test targets
The test targets should be pathogenic fungi that are easy to produce spores and germinate, have consistent genetics and maturity, and are easy to examine under a microscope, such as Pyricularia oryzae and Venturia pirina. Record the source of the fungus. 2.2 Instruments and equipment
2.2.1 Centrifuge;
2.2.2 Electronic balance (sensitivity 0.1 mg): 2.2.3 Microscope;
2.2.4 Incubator;
2.2.5 Culture blood;
2.2.6 Counter;
2.2.7 Slide;
2.2.8 Concave slide;bzxz.net
2.2.9 Pipette or pipette, etc.
3 Experimental design
3.1 Test material preparation
The pathogenic fungi to be used in the test are cultured on a suitable culture medium, or the diseased tissue is cultured in a moisturizing manner and is used for use after spores are produced. 3.2 Agents
3.2.1 Test Agents
The test agent uses the original drug (mother drug) and indicates the generic name, trade name or code, content and manufacturer. 3.2.2 Control Agents
The control agent uses the original drug of the fungicide that has been registered and commonly used in production to inhibit spore germination. 3.3 Experimental steps
3.3.1 Preparation of spore suspension
The cultured pathogenic fungal spores are eluted from the culture medium or diseased tissue with deionized water, filtered, centrifuged (1000r/min) for 5min, the supernatant is discarded, deionized water is added, and centrifuged again. Finally, the spores are resuspended in deionized water to 1×105-1×107 spores per ml, and 0.5% glucose solution is added.
3.3.2 Preparation of agents
NY/T1156.1—2006
Water-soluble agents are directly dissolved and diluted with water. For other agents, use a suitable solvent (such as methanol, acetone, dimethylformamide or dimethylformamide) to dissolve them and dilute them with 0.1% Tween 80 aqueous solution. According to the activity of the agent, set 5 to 7 series of mass concentrations, and the final content of organic solvents shall not exceed 2%.
3.3.3 Agent treatment
Use a pipette or a pipette to draw 0, 5 of the drug solution from low concentration to high concentration, and then draw 0.5mL of the prepared spore suspension to mix the drug solution and spore suspension in equal amounts. Use a micropipette to draw the above mixed solution and drop it onto a concave glass slide, and then place it on a rack with a shallow Layer of water in the culture medium, cover to keep moist and culture in an incubator at a suitable temperature. Each treatment should be repeated at least 3 times, and a treatment without drug should be set as a blank control. 4 Investigation
When the spore germination rate of the blank control reaches more than 90%, check the spore germination of each treatment, randomly observe more than 3 fields of view for each treatment and each repetition, and investigate the total number of spores of no less than 200, and record the number of germination and the total number of spores respectively. The spore germ tube length greater than the short radius of the spore is considered to have germinated. This test should also observe and record abnormal germ tube growth, the number of attachment cells formed, etc. 5 Data statistics and analysis
5.1 Calculation method|| tt||According to the survey data, calculate the relative inhibition rate of spore germination of each treatment in percentage (%). Calculate according to formula (1) and (2), and keep two decimal places after the calculation result:
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Where:
R—spore germination rate:
Number of spore germination:
Total number of spores investigated.
Where:
—Treatment correction spore germination rate;
Treatment spore germination rate;
Ro——blank control spore germination rate.
Where:
Spore germination Relative inhibition rate;
Ro——spore germination rate of blank control;
R——spore germination rate corrected by treatment.
5.2 Statistical analysis
++++++++*(3)
According to the logarithmic value of each agent concentration and the corresponding probability value of spore relative inhibition rate, regression analysis was performed to calculate the ECSECo value of each agent and its 95% confidence limit.
When conducting the combined toxicity test of agents, the synergistic coefficient (SR) or co-toxicity coefficient (CTC) of the mixture was calculated according to the Wedly method or Sun Yunpei method to evaluate the combined action type of the mixture. 2
NY/T1156.1—2006
Wadley method: The synergistic effect of the mixed use of agents was evaluated based on the synergistic coefficient (SR), that is, SR<0.5 is an antagonistic effect, 0.5≤SR1.5 is an additive effect, and SR>1.5 is a synergistic effect. Calculate the synergistic coefficient (SR) according to formula (4) and (5): PA+PB
X=PA/A+Pa/B
Wherein:
Wherein:
Theoretical ECso value of the mixture, in milligrams per liter (mg/): The percentage of A in the mixture;
The percentage of B in the mixture;
The ECso value of A in the mixture, in milligrams per liter (mg/L): The EC value of B in the mixture, in milligrams per liter (mg/L). SR
The synergistic coefficient of the mixture;
Theoretical ECso value of the mixture, in milligrams per liter (mg/L);X1—The measured ECso value of the mixture, in milligrams per liter (mg/L). (4)
Sun Yunpei method: The synergistic effect of mixed drugs is evaluated based on the co-toxicity coefficient (CTC value), that is, CTC ≤ 80 is antagonistic, 80
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