CN202166640U - Kit and coated glass tube for rapid detection of insecticide toxicity to whitefly - Google Patents

Abstract

The utility model "a kit and a glass tube with drug film for quickly detecting the toxicity of insecticides to whitefly" belongs to the detection technology of insecticide toxicity. The kit is characterized in that it includes at least one A glass tube with an open end attached to the pesticide film with the insecticide to be tested as the main component. superior. With the advantages of small error, high efficiency, and strong stability, it is specially designed for the rapid determination of the toxicity activity of insecticides on whitefly adults.

Description

Kit and coated glass tube for rapid detection of insecticide toxicity to whitefly

technical field

The utility model relates to an insecticide toxicity detection technology, in particular to a kit and a drug-coated glass tube for rapidly detecting the toxicity of an insecticide to whitefly.

Background technique

The whitefly Bemisia tabaci belongs to the order Hemiptera, the family Amisidae, and the genus Amisia. Found on Greek tobacco in 1889, it was named whitefly Aleyrodes tabaci (Gennadius) (Gennadius, 1889). In 1894, it was found on sweet potatoes in Florida, USA, and identified as Bemisia inconspicua. At present, Bemisia tabaci has been widely distributed in more than 100 countries and regions except Antarctica, including tropical, subtropical and adjacent temperate regions. In China, the current survey found that it is distributed in more than 20 provinces and regions such as Guangdong, Guangxi, Hainan, Fujian, Yunnan, Shanghai, Zhejiang, Jiangxi, Hubei, Sichuan, Shaanxi, Beijing, Taiwan, Xinjiang, Hebei, and Shandong.

Bemisia tabaci is a worldwide economic pest that causes serious damage to agricultural, horticultural and ornamental crops (Chen Wenxiong and Zhang Huanying, 1997; Luo Zhiyi et al., 1989; Castle et al., 2002; Bacci et al., 2007Ahmad, 2007; Dinsdale et al., 2010), on vegetable crops, it mainly damages cucumber, tomato, eggplant, cowpea, vegetable soybean, kale and cauliflower. The insect has a wide range of hosts, strong fecundity, and serious overlapping of generations. It damages host plants by sucking adults and nymphs on the back of leaves, which can cause more than 50% yield reduction; the honeydew produced by Bemisia tabaci is also a coal pollution disease. The most important thing is that whitefly is also the vector of various plant virus diseases. It has been confirmed that more than 100 plant viruses are transmitted by whitefly. Especially in recent years, the tomato yellow leaf curl virus disease (TYLCD), which has been highlighted in production from south to north in China and has caused the failure of tomato sheds in some areas, is caused by the transmission of Bemisia tabaci.

At present, the control of whitefly in production mainly relies on chemical pesticides. However, adult whitefly has a certain ability to fly and has a strong ability to evade spraying of pesticides. It is one of the most difficult pests to control in production. With the use of insecticides, the whitefly has gradually developed resistance to more and more different types of insecticides, and even due to the selection pressure of a single insecticide, the biotype of the whitefly is also changing. Succession has occurred, and some areas have changed from type B to type Q. Because type Q is more resistant to insecticides and has stronger biological characteristics, it is more challenging to control this insect in production. .

Bifenthrin is a high-efficiency synthetic pyrethrin insecticide and acaricide. It has contact and stomach poisoning effects, no systemic and fumigation effects, a wide insecticidal spectrum, good control effect on mites, and rapid poisoning effect . It does not move in the soil, is relatively safe for the environment, and has a long residual effect period. It is suitable for various Lepidoptera larvae, whiteflies, aphids, and herbivorous leaves on various crops including eggplant, cabbage, cauliflower, and lettuce. Mite control. This chemical agent has entered the market since the 1980s, and it still plays its preventive role in production. According to the test results of the resistance of whiteflies in different areas, the resistance to bifenthrin of whiteflies in many areas is high at present, and the sensitivity is not high, so the control effect is significantly reduced compared with before.

The traditional method for detecting the sensitivity of pests to insecticides is realized by indoor virulence testing, and the indoor virulence testing of different pests varies with different pest species. The sensitivity test of Bemisia tabaci to bifenthrin and other pyrethrin insecticides is mainly carried out by agar leaf dipping method (Feng YT et al., 2010). However, this assay method needs to consume 2-3 days, and the degree of toxicity of the agent can still be detected. In addition, this method requires certain test consumables (such as finger tubes, insect-free seedlings, punchers, etc.), and the breeding of insect-free seedlings needs to spend a lot of manpower and financial resources; The operation is relatively simple and time-consuming, but because adults of Bemisia tabaci have the ability to fly and are small in size, they are easily caught between the filter paper membrane and the carrier and cause damage to varying degrees, especially when the wings are damaged. When it comes to the inspection and recording of the test results, a large test error is caused. Therefore, establishing and optimizing a new rapid detection method for pesticides has important guiding significance for evaluating the toxicity of pesticides to pests in a short period of time, so as to select and apply pesticides in a targeted manner.

Utility model content

According to the needs of the above-mentioned fields, the utility model provides a kit for detecting the toxicity of insecticides to whitefly and a drug-coated glass tube, which has the advantages of small error, high efficiency, strong stability, etc. Toxic activity of insecticides against adults of whitefly whitefly.

The kit for detecting the toxicity of insecticides to whitefly tabaci is characterized in that it comprises a glass tube with an opening at one end with a drug film mainly composed of the insecticide to be tested attached to the inner wall, and the drug film is attached to the glass tube On the inner wall of the bottom of the tube to one-third to one-half of the height of the glass tube.

The drug film is attached to the inner wall at the bottom of the glass tube to a third of the height of the glass tube.

The kit includes 5 to 7 groups of glass tubes with insecticide film attached to the inner wall and one end of the opening, and the effective content of the insecticide in the film between the glass tubes of each group is gradient.

The glass tube is a finger tube.

The insecticide refers to bifenthrin.

The drug-coated glass tube for detecting the toxicity of insecticides to whitefly tabaci is characterized in that the drug film is attached to the inner wall of the glass tube from the bottom of the glass tube to one-third to one-half of the height of the glass tube.

The utility model provides an insecticide virulence detection kit specially for whitefly adults on the basis of the traditional glass tube drug film method and filter paper drug film method, combined with the biological characteristics of whitefly adults who are good at flying. It includes at least one glass tube with an open end on the inner wall of which a drug film with the insecticide to be tested as the main component is attached. The main improvement of the utility model is that the drug film does not completely cover the inner wall of the glass tube, but only covers the bottom of the glass tube to the height of one-third to half of the above, which can reduce the amount of chemicals used to a certain extent, thereby reducing the cost of pesticides. Pollution and residues; in use, when adult bemisia tabaci are inserted into the pipe, a section of the opening needs to be blocked with a cotton plug, and the tampon is plugged to the upper edge of the drug film, so that adults bemisia tabaci can move freely and their range of activities is limited. In a drug environment; such a design can moderately increase the concentration of insecticides, and quickly detect the impact of the tested insecticides on whitefly adults in just 3 hours or more (according to different characteristics of insecticides). The level of virulence of the whitefly; this method can quickly determine the degree of resistance of the whitefly population to insecticides in different areas on the basis of the determination of the sensitive whitefly population, and provide a basis for the use of chemical agents in the field to control the insecticide. Provide timely and rapid drug selection when pests are encountered.

The kit of the utility model is used to detect the toxicity activity of a kind of insecticide to whitefly, which requires a relatively wide range of dosage range of the insecticide. Those skilled in the art can set several groups of drug-coated glass The doses of insecticides between each coated glass tubes were changed in a gradient, so as to accurately find out the lethal dose of an insecticide for different populations of whitefly. Considering cost saving and detection efficiency, the utility model is preferably provided with 5 to 7 groups of drug-coated glass tubes. One or more can be set for each group.

The glass tube used in the utility model is preferably a commonly used finger tube.

In the preferred implementation of the present utility model, the insecticide refers to bifenthrin. It was found that whiteflies in some areas have higher drug resistance to bifenthrin, and the sensitivity is not high. The control effect of the traditional dosage is the same as before. followed by a significant decrease. To detect the toxicity of bifenthrin to whitefly in different regions has very practical significance for the effective use of this agent for effective control of whitefly.

In summary, the kit of the present utility model can be used as a rapid qualitative detection of insect sensitivity to chemicals, and the established diagnostic dose LC ₆₀ (40.6644mg/L) sensitive to bifenthrin can quickly evaluate tobacco powder in different regions. A practical approach to insecticide (bifenthrin) resistance in lice. The kit can quickly establish the diagnostic dosage of more pesticides for pests, so as to guide the rapid selection and application of pesticides when chemical pesticides are controlled in production when pests occur in large numbers.

Description of drawings

Fig. 1. schematic diagram of the Chinese medicinal film glass tube of the utility model kit,

Among them: 1-glass tube, 2-drug film, 3-the upper edge of the drug film.

Figure 2. Stability test of glass tube drug film at different temperatures and storage times.

Detailed ways

Embodiment 1. Preparation and use method of kit

1. Tested insects and feeding

Bemisia tabaci was collected from the cabbage field in the northern field of Vegetable and Flower Research Institute, Chinese Academy of Agricultural Sciences in October 2004. It was identified as type B Bemisia tabaci by mtCOI (mitochondrial cytochrome oxidase I gene) molecular marker, and named as SS population. It has been raised on insect-free 'Jingfeng No. 1' cabbage (Brassica oleracea L., cv Jingfeng 1) seedlings until now.

The biotype of the bifenthrin-resistant Bemisia tabaci population was also type B, named LB-RR population. The bioassay by agar leaf dipping method in the laboratory found that its resistance multiple was about 13.9983 times relative to the SS population. Insect-free cabbage seedlings were subcultured indoors and sprayed with corresponding insecticides for resistance selection.

2. Pesticides

95.6% bifenthrin technical, produced by Jiangsu Changlong Chemical Co., Ltd.

3. Traditional Agar Leaf Dip Method Bioassay

Spread about 2mL of 1.8%-2% agar on the bottom of the finger tube. After the agar solidifies, wipe off the water vapor on the tube wall with absorbent paper. Harvest the flat cabbage leaves without pesticide application, and use a puncher to punch out leaf discs with a diameter of 22 mm to avoid thick leaf veins. Set 5-7 drug treatment concentrations and 1 group of blank controls, set 4 repetitions for each concentration, and prepare distilled aqueous solution containing 0.125‰ Triton X-100 with water for dilution. After preparing the medicinal solution, soak each leaf disc for 10 seconds, dry it and stick it flat on the bottom of the tube facing the agar. With the mouth of the tube facing down, lightly pat the leaves with whiteflies on them to let whiteflies fly in freely. 20 to 35 heads per tube are inserted into 2/3 of the tube length with cotton plugs, and the distance from the leaves is not less than 15mm. Put the finger tube upside down in the light incubator with the agar facing upwards, set the temperature at 25°C, and the photoperiod 14h:8h (L:D), and check the number of dead insects in each tube after 48h. The total number of concentrations and the number of deaths were analyzed using the PBT data processing system developed by Wuhan Vegetable Science Research to analyze the death ratio of whiteflies within a certain period of time under the treatment of each pesticide concentration, and the corresponding LC ₅₀ , LC ₉₀ , regression equation, 95 % Confidence limit, standard deviation, chi square value and correlation coefficient, etc.

4. The utility model glass tube drug film bioassay method

Weigh 95.6% of the calculated mass of the original medicine of bifenthrin and dissolve it with acetone and double-gradiently dilute it into 6 different concentrations of medicine solutions. At the same time, set an acetone blank control, and set 4 repetitions for each treatment, a total of 28 medicine-coated glasses tube (Figure 1). Use a pipette gun to draw 500 μL of liquid medicine into the bottom of the tube, carefully tilt and rotate the finger tube, the liquid medicine will be fixed on the inner wall of glass tube 1 with the volatilization of acetone, so that it is about 1/3 of the length of the tube from the bottom of the tube A drug film 2 is formed inside, and the amount of drug per m ² of the drug film part of the glass tube is about 9.34mg. After the drug film inside the glass tube is placed until it is completely dry (more than 2h), the insect inoculation is performed. Gently pat the plant to drive the adults of whitefly into the pipe, and the cotton plug is pushed into the upper edge 3 of the medicine film 2 . After treatment, place the finger tube (glass tube 1) upside down in an incubator at 25±1°C, and check the results after 3 hours (depending on the characteristics of the insecticide, the observation time can be extended appropriately).

This experiment shows that the rapid knockdown effect of bifenthrin can cause Bemisia tabaci to show symptoms such as turning over, flapping wings, rolling and unable to fly normally. As the result of the quick test is only 3 hours later (or longer), the effect of the drug film is too late to completely kill the downed whitefly, so the judgment standard used in the inspection is that the whitefly has movement disorders (including complete death and the presence of inability to Movement disorders such as crawling and inability to fly).

For the indoor type B sensitive population (S), the above-mentioned steps were used for bioassay, and three consecutive bioassays with the closest LC ₅₀ were selected for synthesis, and then processed with Probit software, and the concentrations corresponding to LC ₅₀ , LC ₆₀ , and LC ₈₀ were selected as Alternative concentrations for diagnostic doses. As shown in Table 1.

Table 1 Preliminary screening concentrations of bifenthrin for diagnostic doses of whitefly and their respective corrected mortality

It can be seen from Table 1 that the corrected mortality rate of LC ₆₀ is most in line with the expected 60% mortality rate, and the indicator is the most stable. In addition, the corresponding concentration of LC ₈₀ is relatively high. The glass tube coating method is used in the hot summer season, and the coating is easy to absorb moisture and change. Tide, so as to stick to the wings of whitefly adults, which brings inconvenience to the actual operation and brings inevitable errors to the experiment. Therefore, LC ₆₀ (40.6644mg/L) was finally determined as the diagnostic dose.

Example 2. Determination and verification of the diagnostic dosage of insecticides for toxicity to whitefly

1 Determination of diagnostic dose

Adopt the test kit of embodiment 1 and its use and statistical method, take indoor sensitive bemisia tabaci population as the test insect, carry out the sensitivity detection of whitefly adults to different agents, carry out 3-4 tests continuously respectively, wait for the result to be stable Finally, the three data with the most consistent experimental results were combined to calculate the different dosages such as LC ₅₀ , LC ₆₀ , LC ₇₀ , and LC ₈₀ .

Under the conditions of different concentrations calculated above, the above-mentioned dosages were verified on the sensitive Bemisia tabaci SS population, and the experiment was repeated 3-4 times to calculate the corrected mortality. The dose with the most stable results was selected as the diagnostic dose.

In order to verify the diagnostic reliability of LC ₆₀ , three groups of different test insects were treated with the drug film method: the first group, completely indoor B-type bifenthrin-resistant varieties (LB-RR); the second group, completely indoor B type bifenthrin-sensitive variety (SS); the third group, R strain and S strain were mixed at a ratio of 1:1, and controls were set for each treatment group. Combined with the results of the treatment control in each group, the mortality rates of the three treatment groups were analyzed, and the results are shown in Table 2.

Table 2 Stability test of LC ₆₀

It can be seen from Table 2 that the corrected mortality rate of 100% SS population is 58.57%, which is close to 60%, indicating the stability of its diagnosis for SS population. The mortality rate of the LB-RR population treated with LC60 was lower than that of the control treated LB-RR population (3.66%<6.15%), indicating that the treatment of resistant whitefly with this diagnostic dose was almost ineffective and could be equivalent to the blank control. And 50% (LB-RR + SS) mortality rate ≈ 50% (LB-RR mortality rate + SS mortality rate). It shows that LC60 has strong indication and stability as a diagnostic dose.

2 Verification of diagnostic dose

Use indoor sensitive populations and corresponding resistant populations as test insects to verify the final diagnostic dose above. 100% SS population, 50% SS population+50% RR population, and 100% RR population were respectively set up to verify the diagnostic dose. Finally, a field Bemisia tabaci population was set up for further verification.

The above-mentioned diagnostic doses were used in field Bemisia tabaci populations (FF) on greenhouse tomatoes for verification experiments. Still apply LC ₆₀ (40.6644mg/L) to treat three groups of different test insects: the first group, the field population (FF); the second group, the indoor type B bifenthrin sensitive population (SS); the third group, the FF population Mix with SS stock at a ratio of 1:1. In combination with the respective control treatments of the three groups, the glass tube drug film method and the diagnostic dose concentration of bifenthrin were used for determination, and the results are shown in Table 3.

Table 3 The results of the field application of diagnostic doses

Table 3 shows that under the treatment of the diagnostic dose LC60 concentration, the mortality rate of the field FF population (27.06%) is higher than that of the indoor RR resistant population (3.66%), but it is much lower than 60%. There is a certain level of resistance to bifenthrin in the population. Afterwards, the field population was bioassayed by leaf dip bioassay, and it was found that the resistance multiple was 2.85 relative to the sensitive population, and the resistance level was low. At the same time, in this experiment, the mortality rate of the indoor sensitive SS population was 57.20%, still close to 60%. This test is yet another strong demonstration of the reliability of the diagnostic dose.

Embodiment 3. The stability of glass tube drug film under different storage conditions

After the acetone-diluted bifenthrin of the LC60 diagnostic dosage concentration determined in Example 1 is rolled in the glass finger tube within 1/3 of the length from the bottom, it is placed in a fume hood to dry thoroughly for 1 hour, and then stuffed into a tampon. Seal it with a sealing bag, and store the prepared glass tube drug film in the dark at room temperature and 4°C, for 1 day and 3 days at room temperature, and for 1 day, 3 days, 7 days, and 10 days at 4°C. days, 15 days, 20 days and 30 days. Afterwards, the drug film and the blank control were taken out, and after returning to room temperature, the stability of the diagnostic dose on the indoor sensitive Bemisia tabaci SS population was determined by the above method, and each treatment was repeated 3 times.

The glass tube drug film was stored under different temperature and time conditions, and the mortality rate of Bemisia tabaci was measured, and the corrected mortality rate was calculated. The results are shown in Figure 2. It can be seen from Figure 2 that at room temperature and 4°C, the diagnostic dose concentration in the glass tube drug film has good stability, and the corrected mortality rate of the test insects is basically about 60%, especially at 4°C, its stability can be up to a month. The results provide a guarantee for the commercialization of this rapid diagnostic method.

Claims (6)

1. detect the kit of pesticide to the Bemisia tabaci virulence; It is characterized in that; Comprise that being attached with at least one its inwall with pesticide to be measured is the glass tube of an end opening of the medicine film of principal ingredient, said medicine film attached to the pipe of glass tube at the bottom of to the inwall at 1/3rd to 1/2nd places of glass tube height.

2. detection pesticide according to claim 1 is characterized in that the kit of Bemisia tabaci virulence: said medicine film attached to the glass tube pipe at the bottom of to the inwall at 1/3rd places of glass tube height.

3. detection pesticide according to claim 1 is to the kit of Bemisia tabaci virulence; It is characterized in that: said kit comprises the glass tube that is attached with an end opening of pesticide medicine film on 5～7 groups of inwalls, and each effective content of organizing pesticide in the medicine film between the glass tube in gradient.

4. detection pesticide according to claim 1 is characterized in that the kit of Bemisia tabaci virulence: said glass tube is a dactylethrae.

5. according to the kit of the arbitrary described detection pesticide of claim 1～4 to the Bemisia tabaci virulence, it is characterized in that: said pesticide refers to Biphenthrin.

6. detect the medicine film glass pipe of pesticide, it is characterized in that the Bemisia tabaci virulence: said medicine film attached to the pipe of glass tube at the bottom of to the inwall at 1/3rd to 1/2nd places of glass tube height.

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