Hyphantria cunea field population drug resistance detection kit and detection method thereof

Abstract

The invention discloses a field population drug resistance detection kit for fall webworms and a detection method of the field population drug resistance detection kit, and belongs to the field of pest control. The detection kit comprises a plurality of glass culture dishes, artificial hyphantria cunea feed, agar and insecticides, the artificial hyphantria cunea feed and the agar are contained in the glass culture dishes, and each glass culture dish further contains seven concentration gradients corresponding to five insecticides. According to the invention, on the basis of the relationship between sensitive strain hyphantria cunea raised in a laboratory and insecticides, the response relationship between different insecticides and hyphantria cunea larvae is prepared, and the hyphantria cunea larvae in the field are directly used as materials for detection; the detection result can be obtained within 48 hours, the detection time is greatly shortened, the detection efficiency is improved, and the method is suitable for various indoor and outdoor environments and has practical application value.

Description

Technical Field

[0001] The present invention belongs to the field of pest control, and particularly relates to a kit for detecting the insecticide resistance of the wild population of Hyphantria cunea and a detection method thereof. Background Art

[0002] Hyphantria cunea is a highly harmful alien invasive species with strong reproductive ability, wide adaptability and multiple transmission routes, which poses a huge threat to the agricultural and forestry production, ecological environment and urban greening in China. Since its introduction into China, the distribution range of Hyphantria cunea has expanded rapidly and it is now widely distributed in many provinces of China. It mainly damages fruit trees, street trees, ornamental trees, etc., and can feed on the leaves of various plants, causing a large number of leaves to fall from the trees, seriously affecting the growth and ornamental value of the trees, and even leading to the death of the trees. In addition, the large-scale reproduction and spread of Hyphantria cunea will also damage the local ecosystem and affect biodiversity. In the process of controlling Hyphantria cunea, chemical insecticides have always played an important role. However, the long-term and large-scale use of chemical insecticides will not only pollute the environment, but also may cause Hyphantria cunea to develop insecticide resistance, reduce the control effect of insecticides, and increase the control difficulty and cost. At present, the research on the resistance of Hyphantria cunea is relatively less, and the key issues such as the resistance formation mechanism, resistance level and resistance differences among different populations are not clear. In the actual control work, the lack of effective resistance monitoring means and scientific and reasonable drug use guidance leads to certain blindness in the selection and use of insecticides, further aggravating the risk of insecticide resistance. There is an urgent need to develop a rapid, accurate and simple kit for diagnosing the resistance of Hyphantria cunea to monitor the resistance levels of Hyphantria cunea in different regions and different populations to common insecticides, provide a basis for scientifically formulating control strategies, reasonably selecting and using insecticides, effectively delaying and overcoming the development of insecticide resistance of Hyphantria cunea, improving the control effect, and ensuring the safety of agricultural and forestry production and ecological environment in China. Summary of the Invention

[0003] Aiming at the deficiencies of the prior art, the present invention provides a kit for detecting the insecticide resistance of the wild population of Hyphantria cunea and a detection method thereof, which can quickly detect the drug resistance of Hyphantria cunea to a variety of insecticides.

[0004] To achieve the above object, the present invention adopts the following technical solutions: A detection kit for the insecticide resistance of the field population of Hyphantria cunea, comprising a plurality of glass culture dishes, artificial feed for Hyphantria cunea, agar and insecticides. The glass culture dishes contain artificial feed for Hyphantria cunea and agar, and each glass culture dish also contains one of the 7 concentration gradients corresponding to the following 5 insecticides respectively: The concentrations of chlorantraniliprole are: 0.10 mg / L, 0.50 mg / L, 1.00 mg / L, 1.50 mg / L, 2.00 mg / L, 2.50 mg / L, 5.00 mg / L; The concentrations of abamectin are: 0.10 mg / L, 0.50 mg / L, 2.00 mg / L, 5.00 mg / L, 10.00 mg / L, 20.00 mg / L, 50.00 mg / L; The concentrations of chlorbenzuron are: 500.00 mg / L, 1000.00 mg / L, 2000.00 mg / L, 5000.00 mg / L, 10000.00 mg / L, 15000.00 mg / L, 20000.00 mg / L; The concentrations of deltamethrin are: 0.50 mg / L, 1.00 mg / L, 2.50 mg / L, 5.00 mg / L, 10.00 mg / L, 25.00 mg / L, 50.00 mg / L; The concentrations of cyantraniliprole are: 0.50 mg / L, 1.00 mg / L, 2.00 mg / L, 5.00 mg / L, 10.00 mg / L, 20.00 mg / L, 50.00 mg / L.

[0005] Further, the formula of the artificial feed for Hyphantria cunea is as follows: Each component is calculated by its weight percentage: germ 5 - 12, sucrose 2 - 5, protein 3 - 7, Wesson salt 0.3 - 1, sorbic acid 0.4 - 1.2, methyl p - hydroxybenzoate 0.1 - 0.5, ascorbic acid 0.02 - 0.19, vitamin B 0.03 - 0.15, agar 1.1 - 1.8, cholesterol 0.05 - 0.16 and water 75 - 85.

[0006] The present invention also provides a method for detecting the insecticide resistance of the field population of Hyphantria cunea, using the above - mentioned kit, and the steps are as follows:

[0007] S1: Put 1.6 g of agar and 80 mL of water into the sterilized glass culture dish, boil until melted, add 22 g of artificial feed for Hyphantria cunea and stir evenly, then boil again;

[0008] S2: Add insecticides of different concentration types to the glass culture dish in step S1, stir evenly, cool to room temperature, make marks, and store sealed at 4°C for standby;

[0009] S3: The eggs of the indoor population of white moth were fumigated with formaldehyde, and the larvae were raised in a 200 mL transparent plastic insect box. The third-instar larvae of the white moth with good growth status were selected and placed in glass culture dishes containing different concentrations of insecticides. Ten larvae were treated with each concentration, and at least three biological tests were performed; the lids were sealed and marked;

[0010] S4: Place the glass culture dish in an environment of 5°C and relative humidity of 20-30% for 48 hours, and then measure the mortality rate. Use a brush to gently touch the tail of the gypsy moth. If it does not move within 30 seconds, it is considered dead. The control mortality rate is less than 10% for effective determination, and the control mortality rate is used for correction to calculate the sublethal concentration LC of the 3rd instar larvae of the indoor population of gypsy moth 90 , that is, the diagnostic dose of each pesticide is obtained; the sublethal concentration LC of chlorantraniliprole, avermectin, diflubenzuron, deltamethrin and cyantraniliprole on the third instar larvae of the indoor population of the American moth is determined. 90 The wild population is treated to obtain a 48-hour mortality rate. If the mortality rate is ≥90%, the wild population is sensitive to the insecticide; if the mortality rate is <90%, the wild population has developed resistance to the insecticide.

[0011] Furthermore, the LC of the indoor population of the third-instar larvae of the American moth was measured. 50 and wild population LC 90 Based on the mortality rate at the concentration, the resistance ratio is calculated = LC of the wild population 50 Value / Indoor population LC 50 value, and finally obtain the linear equation of "mortality-resistance ratio" of the American white moth population.

[0012] Compared with the prior art, the beneficial effects of the present invention are as follows: the test kit of the present invention is based on the feed mixing method, and the test results can be obtained within 48 hours, which greatly shortens the test time and improves the test efficiency. Specific experimental data show that the test kit is used to detect the 3rd instar larvae of the American white moth within 48 hours, and the determination result of the mortality rate is consistent with the leaf dipping method. The present invention is based on the relationship between the sensitive strain American white moth and the insecticide raised in the laboratory, and the response relationship between different insecticides and American white moth larvae is prepared. The American white moth larvae in the field are directly used as materials for detection, and it has the advantages of simple operation, time saving, and foresight. It can be used as a method for early resistance monitoring, and the resistance level of the American white moth to chlorantraniliprole, abamectin, cypermethrin, cyanamide, etc. can be quickly and accurately determined, thereby guiding the rational management of the American white moth in production practice, exploring the degree of resistance formation between different populations, and providing help for the subsequent rational selection of insecticides and efficient prevention and control of American white moth. DETAILED DESCRIPTION

[0013] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0014] Example 1

[0015] A detection kit for the insecticide resistance of the field population of Hyphantria cunea includes multiple glass culture dishes, artificial feed for Hyphantria cunea, agar, and insecticides. The glass culture dishes contain artificial feed for Hyphantria cunea and agar. Each of the glass culture dishes also contains one of the 7 concentration gradients corresponding to the following 5 insecticides: the concentrations of chlorantraniliprole are respectively: 0.10 mg / L, 0.50 mg / L, 1.00 mg / L, 1.50 mg / L, 2.00 mg / L, 2.50 mg / L, 5.00 mg / L; the concentrations of abamectin are respectively: 0.10 mg / L, 0.50 mg / L, 2.00 mg / L, 5.00 mg / L, 10.00 mg / L, 20.00 mg / L, 50.00 mg / L; the concentrations of chlorbenzuron are respectively: 500.00 mg / L, 1000.00 mg / L, 2000.00 mg / L, 5000.00 mg / L, 10000.00 mg / L, 15000.00 mg / L, 20000.00 mg / L; the concentrations of deltamethrin are respectively: 0.50 mg / L, 1.00 mg / L, 2.50 mg / L, 5.00 mg / L, 10.00 mg / L, 25.00 mg / L, 50.00 mg / L; the concentrations of cyantraniliprole are respectively: 0.50 mg / L, 1.00 mg / L, 2.00 mg / L, 5.00 mg / L, 10.00 mg / L, 20.00 mg / L, 50.00 mg / L. The formula of the artificial feed for Hyphantria cunea is as follows: each component is calculated by its weight percentage: germ 5 - 12, sucrose 2 - 5, protein 3 - 7, Wesson salt 0.3 - 1, sorbic acid 0.4 - 1.2, methyl p-hydroxybenzoate 0.1 - 0.5, ascorbic acid 0.02 - 0.19, vitamin B 0.03 - 0.15, agar 1.1 - 1.8, cholesterol 0.05 - 0.16, and water 75 - 85. This formula optimizes the raw materials and additives according to the needs of Hyphantria cunea and can fully meet the nutritional elements required for the growth of Hyphantria cunea larvae. The eggs of the indoor population of Hyphantria cunea are purchased from the Research Institute of Forest Ecology, Environment and Nature Conservation, Chinese Academy of Forestry.

[0016] The rapid detection kit for insecticide resistance of the gypsy moth of the present invention contains a variety of original pesticides, so that there are multiple options for detecting the insecticide resistance of the gypsy moth in the forest, or for identifying multiple insecticides. The diagnostic dose of the insecticide is determined by testing the toxicity of different concentrations of insecticides on the 3rd instar larvae of the indoor population of the gypsy moth in the special glass culture dish, and calculating the LC 90 As the diagnostic dose of insecticide for insects of this age.

[0017] The diagnostic dose of the pesticide is determined by the following steps:

[0018] (1) Preparation of glass culture dishes for rapid identification of insecticide resistance of the American white moth

[0019] Take a glass culture dish and sterilize it at 121℃ in an autoclave for 20 min.

[0020] (2) Preparation of medicine:

[0021] Step 1: Weigh 22g of artificial diet for the gypsy moth and 1.6g of agar. Put the agar into 80mL of water and boil it in a microwave oven until it melts. Add the diet and stir evenly. Boil it in a microwave oven and cool it to room temperature.

[0022] Step 2: Prepare different concentrations of pesticides, and prepare the stock solution of each concentration of pesticide. The pesticides include 7 concentrations of chlorantraniliprole (0.10mg / L, 0.50mg / L, 1.00mg / L, 1.50mg / L, 2.00mg / L, 2.50mg / L, 5.00mg / L), avermectin (0.10mg / L, 0.50mg / L, 2.00mg / L, 5.00mg / L, 10.00mg / L, 20.00mg / L, 50.00mg / L), diflubenzuron (500.00mg / L, 1000.00mg / L, 2000.00mg / L, 50 00.00mg / L, 10000.00mg / L, 15000.00mg / L, 20000.00mg / L), cypermethrin (0.50mg / L, 1.00mg / L, 2.50mg / L, 5.00mg / L, 10.00mg / L, 25.00mg / L, 50.00mg / L), cyantraniliprole (0.50mg / L, 1.00mg / L, 2.00mg / L, 5.00mg / L, 10.00mg / L, 20.00mg / L, 50.00mg / L).

[0023] Step 3: Add the medicine into the uncooled and solidified feed, stir evenly, and divide into glass culture dishes.

[0024] Step 4: When the temperature drops to room temperature, mark it, put it in a sealed bag and store it in a 4℃ refrigerator for later use.

[0025] (3) Preparation of indoor population of gypsy moth

[0026] The eggs and artificial diets of the indoor population of American white moth were purchased from the Institute of Forest Ecology, Environment and Nature Conservation, Chinese Academy of Forestry. The egg masses of the indoor population were fumigated with 10% formaldehyde solution, and the larvae were raised in 200mL transparent plastic insect boxes under the following conditions: (25±1)℃, relative humidity of (20-30)%, and photoperiod of 16L:8D. The artificial diet formula of the American white moth was purchased from the Institute of Forest Ecology, Environment and Nature Conservation, Chinese Academy of Forestry.

[0027] (4) Toxicity assay

[0028] The third instar larvae of the gypsy moth in good growth condition were selected and placed in glass culture dishes with medicated feed of different concentrations. Ten larvae were treated with each concentration, and biological measurements were performed at least three times. The lid was sealed and marked, and the culture dishes were placed in an environment of (25±1)℃ and relative humidity of (20-30)%. After 48 hours of feeding, the mortality rate was determined. The tail of the gypsy moth was gently touched with a brush. If it did not move within 30 seconds, it was considered dead. The control mortality rate was less than 10% for effective measurement, and the control mortality rate was used for correction. The data was processed using DPS software to calculate LC 90 , that is, the diagnostic dose of each agent is obtained. This diagnostic dose can be used to quickly detect the resistance level of the wild population of the American white moth to chlorantraniliprole, abamectin, diflubenzuron, cypermethrin, and cyantraniliprole. The kit is used to measure the LC of the indoor population 50 and LC of each wild population 90 Based on the mortality rate at the concentration, the resistance ratio is calculated = LC of the wild population 50 Value / Indoor population LC 50 value, and finally obtain the linear equation of "mortality-resistance ratio" of the American white moth population.

[0029] Example 2

[0030] A method for determining the insecticide resistance of the wild population of the American white moth by leaf dipping method, using a plurality of glass culture dishes, acetone, mulberry leaves, and insecticides. The insecticides include chlorantraniliprole (96.00%), avermectin (76.90%), diflubenzuron (95.18%), deltamethrin (98.20%), and cyantraniliprole (94.00%).

[0031] The diagnostic dose of the pesticide is determined by the following steps:

[0032] (1) Preparation of glass culture dishes for rapid identification of insecticide resistance of the American white moth: sterilize the glass culture dishes in an autoclave at 121°C for 20 min and set aside for use.

[0033] (2) Preparation of mulberry leaves with pesticides:

[0034] Step 1: Select fresh mulberry leaves of similar size, wash them with distilled water, and air-dry.

[0035] Step 2: Prepare pesticide doses of different concentrations and prepare the stock solutions for each concentration. The pesticides include seven concentrations of chlorantraniliprole (0.01 mg / L, 0.05 mg / L, 0.10 mg / L, 0.15 mg / L, 0.20 mg / L, 0.25 mg / L, 0.50 mg / L), abamectin (0.01 mg / L, 0.05 mg / L, 0.20 mg / L, 0.50 mg / L, 1.00 mg / L, 2.00 mg / L, 5.00 mg / L), chlorbenzuron (50.00 mg / L, 100.00 mg / L, 200.00 mg / L, 500.00 mg / L, 1000.00 mg / L, 1500.00 mg / L, 2000.00 mg / L), deltamethrin (0.05 mg / L, 0.10 mg / L, 0.25 mg / L, 0.50 mg / L, 1.00 mg / L, 2.50 mg / L, 5.00 mg / L), and cyantraniliprole (0.05 mg / L, 0.10 mg / L, 0.20 mg / L, 0.50 mg / L, 1.00 mg / L, 2.00 mg / L, 5.00 mg / L).

[0036] Step 3: Put the pesticides into a plastic box, immerse the air-dried mulberry leaves in the pesticides for 15 seconds, and use the mulberry leaves immersed in an equal amount of acetone as the control group. After natural drying, the mulberry leaves with pesticides are obtained.

[0037] Step 4: Put the mulberry leaves with pesticides into a glass petri dish, pack them with a sealed bag, and store them in a 4°C refrigerator for later use.

[0038] (3) Preparation of the indoor population of Hyphantria cunea:

[0039] The eggs of the indoor population of Hyphantria cunea are purchased from the Research Institute of Forest Ecology, Environment and Nature Conservation, Chinese Academy of Forestry. The egg masses of the indoor population are fumigated and disinfected with 10% formaldehyde solution, and the larvae are raised in a 200 mL transparent plastic insect rearing box. The rearing conditions are: (25 ± 1)°C, relative humidity (20 - 30)%, and photoperiod 16L:8D.

[0040] (4) Toxicity determination:

[0041] Select the well - growing 3rd - instar larvae of Hyphantria cunea (Hubei Dawu population, Liaoning Tieling population, indoor population) and transfer them into glass petri dishes with mulberry leaves containing different concentrations of pesticides. Each concentration is treated with 10 larvae, and at least 3 biological assays are carried out. Seal the petri dishes, make good marks, and place the petri dishes in an environment with a temperature of (25 ± 1)°C and a relative humidity of (20 - 30)%. After raising for 48 h, measure the mortality rate. Gently touch the tail of the Hyphantria cunea larva with a brush. If it does not move within 30 s, it is considered dead. When the control mortality rate is less than 10%, the assay is valid, and the data is corrected using the control mortality rate. The data is processed using DPS software. The results of the toxicity assays of 5 pesticides against Hyphantria cunea are shown in Tables 1 to 5, and calculate LC 90 , that is, the diagnostic dose of each pesticide is obtained.

[0042] Table 1 Toxicity of Chlorantraniliprole against 3rd - instar larvae of Hyphantria cunea from different geographical populations

[0043]

[0044] Note: Under the same bioassay method, resistance ratio = LC value of field population / LC value of indoor population. 50 value / LC 50 value of indoor population.

[0045] The 48 - h LC 90 values of Chlorantraniliprole against 3rd - instar larvae of Hyphantria cunea in the indoor population, Tieling population, and Dawu population are 4.91 mg / L, 15.30 mg / L, and 18.40 mg / L respectively. The resistance ratios of the Tieling and Dawu populations to the indoor population are 14.54 times and 15.54 times respectively.

[0046] Table 2 Toxicity of Cyantraniliprole against 3rd - instar larvae of Hyphantria cunea from different geographical populations by leaf - dipping method

[0047]

[0048] The 48 - h LC 90 values of Cyantraniliprole against 3rd - instar larvae of Hyphantria cunea in the indoor population, Tieling population, and Dawu population are 4.94 mg / L, 12.40 mg / L, and 26.28 mg / L respectively. The resistance ratios of the Tieling and Dawu populations to the indoor population are 6.26 times and 11.36 times respectively.

[0049] Table 3 Toxicity of Abamectin against 3rd - instar larvae of Hyphantria cunea from different geographical populations by leaf - dipping method

[0050]

[0051]

[0052] The 48 - h LC of Abamectin against 3rd - instar larvae of Hyphantria cunea in the indoor population, Tieling population, and Dawu population90 They were 2.54 mg / L, 31.18 mg / L, and 53.67 mg / L respectively. The resistance ratios of the Tieling and Dawu populations to the indoor population were 16.23 times and 14.28 times respectively.

[0053] Table 4 Toxicity of chlorbenzuron to the 3rd instar larvae of Hyphantria cunea from different geographical populations determined by leaf-dipping method

[0054]

[0055] The 48h LC of chlorbenzuron to the 3rd instar larvae of Hyphantria cunea from the indoor population, Tieling population, and Dawu population 90 They were 3455.91 mg / L, 11009.16 mg / L, and 12886.62 mg / L respectively. The resistance ratios of the Tieling and Dawu populations to the indoor population were 2.67 times and 4.93 times respectively.

[0056] Table 5 Toxicity of deltamethrin to the 3rd instar larvae of Hyphantria cunea from different geographical populations determined by leaf-dipping method

[0057]

[0058] Note: Under the same bioassay method, resistance ratio = LC of field population 50 value / LC of indoor population 50 value.

[0059] The 48h LC of deltamethrin to the 3rd instar larvae of Hyphantria cunea from the indoor population, Tieling population, and Dawu population 90 They were 4.21 mg / L, 38.78 mg / L, and 17.66 mg / L respectively. The resistance ratios of the Tieling and Dawu populations to the indoor population were 15.74 times and 13.18 times respectively.

[0060] Example 3

[0061] Use the kit in Example 2 to quickly identify the insecticide resistance of the 3rd instar larvae of Hyphantria cunea in the field population (Dawu population, Tieling population, indoor population). The specific method is as follows: The tested populations of Hyphantria cunea are: Tieling population in Liaoning Province, Dawu population in Hubei Province, and indoor population. The 3rd instar larvae of the three populations of Hyphantria cunea are respectively picked into glass petri dishes with medicated feed at different concentrations in the kit. 10 larvae are treated at each concentration, with 3 replicates. Seal the covers and make marks. Place the petri dishes in an environment with a temperature of (25 ± 1) °C and a relative humidity of (20 - 30) % and raise them for 48h, then measure the mortality rate. Gently touch the tail of the Hyphantria cunea body with a brush. If it does not move within 30 seconds, it is considered dead. If the mortality rate ≥ 90%, the population is sensitive to the insecticide; if the mortality rate < 90%, the population has developed insecticide resistance.

[0062] Using the kit to measure the LC of the indoor population 50and each wild population LC 90 Based on the mortality rate at each concentration, the resistance ratio was calculated (wild population LC 50 value / laboratory population LC 50 value). Populations with a resistance ratio greater than 1 were resistant populations. The identification results are shown in Table 6:

[0063] Table 6 Resistance diagnosis of chlorantraniliprole against the 3rd instar larvae of Hyphantria cunea from different geographical populations

[0064]

[0065] Note: Under the same bioassay method, the resistance ratio = wild population LC 50 value / laboratory population LC 50 value.

[0066] The 48h LC 90 values of chlorantraniliprole against the 3rd instar larvae of Hyphantria cunea in the laboratory population, Tieling population, and Dawu population were 0.64 mg / L, 21.50 mg / L, and 12.09 mg / L respectively. The resistance ratios of the Tieling and Dawu populations to the laboratory population were 4.31 - fold and 6.44 - fold respectively.

[0067] Compared with the leaf - dipping method for determining the insecticide resistance of wild populations of Hyphantria cunea in Example 2, using the kit of the present invention to diagnose the insecticide resistance of Hyphantria cunea requires less amount of insects, can simply and quickly detect the insecticide resistance of wild populations, is easy to operate, has accurate results, strong applicability, and can truly simulate the insecticide exposure situation of pests during natural feeding.

Claims

1. A kit for detecting insecticide resistance of wild populations of cuneiform moth, comprising a plurality of glass culture dishes, artificial feed for cuneiform moth, agar and insecticide, wherein the glass culture dishes contain artificial feed for cuneiform moth and agar, and characterized in that: Each of the glass culture dishes also contains one of the seven concentration gradients corresponding to the following five pesticides: The concentrations of chlorantraniliprole are: 0.10mg / L, 0.50mg / L, 1.00mg / L, 1.50mg / L, 2.00mg / L, 2.50mg / L, 5.00mg / L; The concentrations of avermectin were: 0.10 mg / L, 0.50 mg / L, 2.00 mg / L, 5.00 mg / L, 10.00 mg / L, 20.00 mg / L, 50.00 mg / L; The concentrations of diflubenzuron are: 500.00mg / L, 1000.00mg / L, 2000.00mg / L, 5000.00mg / L, 10000.00mg / L, 15000.00mg / L, 20000.00mg / L; The concentrations of deltamethrin were: 0.50 mg / L, 1.00 mg / L, 2.50 mg / L, 5.00 mg / L, 10.00 mg / L, 25.00 mg / L, 50.00 mg / L; The concentrations of cyantraniliprole are: 0.50mg / L, 1.00mg / L, 2.00mg / L, 5.00mg / L, 10.00mg / L, 20.00mg / L, and 50.00mg / L respectively.

2. The kit according to claim 1, characterized in that: The formula of the artificial feed for the American white moth is as follows: the components are calculated by weight percentage: 5-12% germ, 2-5% sucrose, 3-7% protein, 0.3-1% Weisse salt, 0.4-1.2% sorbic acid, 0.1-0.5% methyl parahydroxybenzoate, 0.02-0.19% ascorbic acid, 0.03-0.15% vitamin B, 1.1-1.8% agar, 0.05-0.16% cholesterol and 75-85% water.

3. A method for detecting insecticide resistance of field populations of gypsy moth obtained by the kit according to claim 1 or 2, characterized in that: Here are the steps: S1: Put 1.6g agar and 80mL water into a sterilized glass petri dish, boil until melted, add 22g artificial feed of the gypsy moth and stir well, then boil again; S2: Add different concentrations of pesticides to the glass culture dish of step S1, stir evenly, cool to room temperature, mark, seal and store at 4°C for later use; S3: The eggs of the indoor population of gypsy moth were fumigated with formaldehyde, and the larvae were raised in a 200 mL transparent plastic insect box. The third-instar larvae of the gypsy moth with good growth were selected and placed in glass culture dishes containing different concentrations of insecticides. Ten larvae were treated with each concentration, and at least three biological tests were performed; Seal the cap and mark it; S4: Place the glass culture dish in an environment of 5°C and relative humidity of 20-30% for 48 hours, and then measure the mortality rate. Use a brush to gently touch the tail of the gypsy moth. If it does not move within 30 seconds, it is considered dead. The control mortality rate is less than 10% for effective determination, and the control mortality rate is used for correction to calculate the sublethal concentration LC of the 3rd instar larvae of the indoor population of gypsy moth 90 , that is, the diagnostic dose of each pesticide is obtained; the sublethal concentration LC of chlorantraniliprole, avermectin, diflubenzuron, deltamethrin and cyantraniliprole on the third instar larvae of the indoor population of the American moth is determined. 90 The wild population is treated to obtain a 48-hour mortality rate. If the mortality rate is ≥90%, the wild population is sensitive to the insecticide; if the mortality rate is <90%, the wild population has developed resistance to the insecticide.

4. A method for detecting insecticide resistance of wild populations of gypsy moth according to claim 3, characterized in that: Based on the measured mortality rates of indoor populations at LC50 and LC90 concentrations of various wild populations, the resistance ratio was calculated as LC50 value of wild populations / LC50 value of indoor populations, and finally the linear equation of "mortality-resistance ratio" of the gypsy moth population was obtained.

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