An entomopathogenic composition for controlling tomato leafminer, its preparation method and application

Abstract

This invention discloses an insect pathogen composition for controlling tomato leafminer, its preparation method, and its application, belonging to the field of agricultural technology. The insect pathogen composition for controlling tomato leafminer of this invention includes a compound agent of symbiotic bacterial fermentation products of insect pathogenic nematodes sprayed on tomato leaves, a compound preparation of insect pathogenic nematodes applied in holes during tomato seedling transplanting, and a simulated tomato leaf-bearing moth-attracting ball for trapping tomato leafminer. This invention also discloses the preparation method of this insect pathogen composition for controlling tomato leafminer and its application in controlling tomato leafminer. This invention can reduce the adult insect population density in greenhouses; reduce the opportunity for female tomato leafminer to lay eggs on leaves and fruits, and reduce the overall egg production in greenhouses, thus reducing the larval population density. This invention achieves its control of tomato leafminer through the synergistic effect of multiple components.

Description

Technical Field

[0001] This invention belongs to the field of agricultural technology, specifically relating to an insect pathogen composition for controlling tomato leafminer, its preparation method, and its application. Background Technology

[0002] The tomato leafminer (Tuta absoluta), belonging to the family Gelechiidae in the order Lepidoptera, is native to Peru, South America, and is a highly destructive leaf-mining pest. Since its initial discovery in Xinjiang, my country in 2017, this insect has rapidly spread to more than 20 provinces (autonomous regions and municipalities) in Southwest, Northwest, East, North, and Central China, with a clear trend of further expansion (Zhang et al., 2021). The tomato leafminer has a wide host range, including more than 40 species of plants in 9 families such as Solanaceae, Fabaceae, Brassicaceae, and Poaceae, but it is most fond of tomatoes and can cause damage throughout the entire growth period of tomato plants (Desneux et al., 2010). The tomato leafminer primarily damages plants as larvae by burrowing into leaves and feeding on the leaf tissue, creating tiny tunnels; it can also bore into fruits, creating holes that lead to premature aging of the plant and fruit rot. Under normal circumstances, the tomato leafminer can reduce tomato yield by 20%–30%, and in severe outbreaks, it can lead to yield losses of 80%–100%, making it one of the most devastating pests in the tomato industry (Desneux et al., 2010; Zhang Guifen et al., 2018). my country has the world's largest tomato planting area and is the world's third-largest tomato producer and the largest exporter of processed tomato products (Food and Agriculture Organization, 2017). The invasion and colonization of the tomato leafminer poses a significant threat to the safe production of my country's tomato industry (Xian Xiaoqing et al., 2019).

[0003] The occurrence and damage of the tomato leafminer are covert. Adults typically lay their eggs on the underside or front of leaves at the top of the plant, or on tender stems. After hatching, the larvae burrow into leaves or fruits to feed. Most larvae develop into mature pupae by spinning silk and descending to a depth of 1-2 cm in the soil (Uchoa-Fernandes 1995; Biondi et al., 2018). Due to its covert occurrence and damage, chemical control is not thorough and frequent application of pesticides is necessary. et al., 1998; Guedesand Long-term use of pesticides can lead to increased resistance (Silva et al., 2016; Guedes et al., 2019; Oke et al., 2020; Li Xiaowei et al., 2022), and has adverse effects on pollinating insects and natural enemies, pollutes the ecological environment, and causes risks to the planting industry, consumers and the environment (Zappalà et al., 2012; Zlof and Suffert, 2012).

[0004] Currently, control measures for tomato leafminer at different stages include: spraying chemical agents such as chlorantraniliprole and methoxyfenozide during the larval stage; and using chemical sex pheromones to attract and kill male adults, reducing their numbers and egg production. Chemical control remains the primary method for controlling tomato leafminer. However, chemical control easily leads to problems such as rapid increases in pesticide resistance, environmental pollution, and the poisoning of non-target insects. Therefore, there is an urgent need for a more effective and environmentally friendly method for controlling tomato leafminer.

[0005] Entomopathogenic nematodes of the genera *Steinernema* and *Heterorhabditis* are a class of biological enemies with strong insecticidal capabilities, a broad insecticidal spectrum, and the potential for large-scale cultivation. These nematodes possess active search capabilities and are highly effective against underground and boring pests (Georgis et al., 2006), and have been widely used in pest control in agriculture, forestry, forage, floriculture, and sanitation (Ehlers, 2001; Gaugler et al., 2002; Labaude and Griffin, 2018). They also show potential for biological control of the tomato leafminer moth, which occurs in a concealed manner. Extensive indoor research has been conducted abroad on tomato leafminer larvae, identifying several nematode species highly toxic to the larvae. However, field techniques for efficiently utilizing entomopathogenic nematodes to control tomato leafminer moths still require optimization. Summary of the Invention

[0006] One of the objectives of this invention is to provide an insect pathogen composition for controlling tomato leafminer, thereby solving the problem of the high damage and difficulty in controlling tomato leafminer in the prior art.

[0007] A second objective of this invention is to provide a method for preparing the insect pathogen composition.

[0008] A third objective of this invention is to provide the application of this insect pathogen composition.

[0009] The fourth objective of this invention is to provide a method for controlling tomato leafminer.

[0010] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0011] The first aspect of this invention discloses an insect pathogen composition for controlling tomato leafminer, comprising a compound of symbiotic bacterial fermentation products of insect pathogenic nematodes sprayed on tomato leaves, a compound preparation of insect pathogenic nematodes applied in holes during tomato seedling transplanting, and a simulated tomato leaf-bearing moth-attracting ball for trapping female tomato leafminers in gestation.

[0012] Each 1L of the compound preparation of symbiotic fermentation products of entomopathogenic nematodes contains 100-400ml of entomopathogenic nematode symbiotic fermentation broth, 25-100g of tomato leafminer plant repellent, 2.5-10g of laccase inhibitor, 0.5-2g of humectant, 1.5-6g of ethoxylated modified polytrisiloxane, 1.5-6ml of green orange peel oil, and water to make up the volume.

[0013] Each 1L of the compound preparation of entomopathogenic nematodes contains 5,000,000–20,000,000 IJs of entomopathogenic nematodes that have been infected and propagated by 5–6 instar larvae of the large wax moth, 0.5–2mol of polyphenol oxidase inhibitor, 2.5–10g of nematode moisturizing agent, and 50–300g of pest attractant, with water added to make up the volume;

[0014] The simulated tomato moth-attracting ball includes a simulated tomato with leaves and a sponge stick impregnated with 1-nonanol and / or ethyl octanoate placed on it. The surface of the simulated tomato with leaves and the leaf surface are coated with strong adhesive.

[0015] In some embodiments of the present invention, each 1L of the compound of symbiotic fermentation products of entomopathogenic nematodes contains 200ml of entomopathogenic nematode symbiotic fermentation broth, 50g of tomato leafminer plant repellent, 5g of laccase inhibitor, 1g of humectant, 3g of ethoxylated polytrisiloxane, 3ml of green orange peel oil, and water to make up the volume.

[0016] Or / and each 1L of the compound preparation of entomopathogenic nematodes contains 10,000,000 IJs of entomopathogenic nematodes that have been infected and propagated by 5-6 instar larvae of the large wax moth, 1mol of polyphenol oxidase inhibitor, 5g of nematode moisturizing agent, and 65.5-262gg of insect attractant, with water to make up the volume; preferably, the insect attractant is 131g;

[0017] Preferably, each 1L of the compound preparation of entomopathogenic nematodes contains the following insect attractants: tomato sauce 10-40g / L, casein 15-60g / L, wheat germ powder 10-40g / L, soybean powder 5-20g / L, sucrose 2.5-10g / L, glucose 4-16g / L, linoleic acid 2.5-10g / L, soybean oil 1.5-6g / L, vitamin C 5-20g / L, and cellulose 10-40g / L;

[0018] More preferably, each 1L of the compound preparation of entomopathogenic nematodes contains the following insect attractants: 20g / L tomato sauce, 30g / L casein, 20g / L wheat germ powder, 10g / L soybean powder, 5g / L sucrose, 8g / L glucose, 5g / L linoleic acid, 3g / L soybean oil, 10g / L vitamin C, and 20g / L cellulose.

[0019] In some embodiments of the present invention, the preparation method of the symbiotic fermentation broth of entomopathogenic nematodes includes the following steps: after disinfecting and grinding the entomopathogenic nematodes in the infecting stage, a small amount of nematode suspension is taken onto an NBTA plate and cultured at 28±0.5℃ in the dark for 3-5 days. Blue-green single colonies are picked and transferred to a new NBTA plate for further culture. The purification is repeated 2-3 times. After purification, single colonies are picked onto LB medium and cultured at 28±0.5℃ and 100-400 rpm for 8-24 hours to prepare a fermentation seed liquid. The seed liquid is inoculated into an Erlenmeyer flask containing LB liquid medium at an inoculation rate of 1-4%. Fermentation is carried out under the above conditions for 24-96 hours. The supernatant is collected by high-speed centrifugation. This fermentation supernatant is the symbiotic fermentation broth of entomopathogenic nematodes.

[0020] Preferably, the fermentation seed liquid is prepared by culturing at 200 rpm for 15 hours.

[0021] Preferably, the seed culture is inoculated into an Erlenmeyer flask containing LB liquid medium at a 2% inoculation rate;

[0022] Preferably, fermentation is carried out for 48 hours, followed by high-speed centrifugation to obtain the supernatant;

[0023] Or / and the tomato leafminer plant repellent includes rosemary powder;

[0024] Or / and the laccase inhibitor includes 2,3-dimercaptopropanol;

[0025] Or / and the moisturizer includes trehalose. In some embodiments of the invention, the polyphenol oxidase inhibitor includes lycopene; or / and the nematode moisturizer includes polyacrylamide.

[0026] In some embodiments of the present invention, the method for preparing entomopathogenic nematodes that have been infected and propagated by 5th-6th instar larvae of the wax moth is as follows: the 5th-6th instar larvae of the wax moth are infected and propagated; the third instar larvae (IJs) of the entomopathogenic nematodes are collected in distilled water using the White trap method and stored in a refrigerator at 4°C for later use.

[0027] The second aspect of the present invention discloses a method for preparing an insect pathogen composition for controlling tomato leafminer, comprising preparing a compound of symbiotic bacterial fermentation products, a compound of insect pathogenic nematodes, and a simulated leafy tomato moth attractant ball;

[0028] The preparation method of the compound agent of symbiotic fermentation product of entomopathogenic nematodes includes the following steps: take the fermentation liquid of entomopathogenic nematodes symbiotic bacteria, tomato leafminer plant repellent, laccase inhibitor, moisturizer, ethoxylated modified polytrisiloxane and green orange oil according to the formula, mix them evenly, and make up the volume with water;

[0029] The compound preparation of entomopathogenic nematodes includes the following steps: take each component of entomopathogenic nematodes, polyphenol oxidase inhibitor, nematode moisturizer and pest attractant according to the formula, mix them evenly, and make up the volume with water;

[0030] Alternatively, mix all components of the insect attractant evenly first, then mix it evenly with the prescribed amounts of entomopathogenic nematodes, polyphenol oxidase inhibitors, and nematode moisturizers, and make up the volume with water.

[0031] The preparation method of the simulated leafy tomato moth-attracting ball includes the following steps: First, immerse the sponge stick in ethyl octanoate or 1-nonanol or a mixture of ethyl octanoate and 1-nonanol, take it out, and let it dry at room temperature until it no longer drips liquid. Then, insert the sponge stick into the top of the simulated leafy tomato.

[0032] In some embodiments of the present invention, when the sponge rod is immersed in a mixture of ethyl octanoate and 1-nonanol, the weight ratio of ethyl octanoate to 1-nonanol is 100:10 to 40, preferably 100:20.

[0033] The third aspect of the present invention discloses the application of an insect pathogen composition for controlling tomato leafminer in the control of tomato leafminer.

[0034] The fourth aspect of the present invention discloses a method for controlling tomato leafminer, which uses the above-mentioned insect pathogen composition.

[0035] In some embodiments of the present invention, a compound agent of symbiotic bacterial fermentation products of entomopathogenic nematodes is sprayed onto tomato leaves at a rate of 15-20 L per acre.

[0036] When transplanting tomato seedlings, the compound preparation of insect pathogenic nematodes is evenly dripped onto the roots of the tomato seedlings. Preferably, the compound preparation of insect pathogenic nematodes is evenly dripped into a circle with a diameter of 10-15cm, with the base of the seedling roots as the center. More preferably, the dripping volume is 20-25ml per seedling.

[0037] The simulated tomato moth-attracting balls with leaves are hung in the tomato field; preferably, the hanging height is 20-40cm from the ground; preferably, 20-30 balls are hung per acre.

[0038] Laccase is an oxidase containing four copper ions and possessing ortho-diphenol oxidation activity. Laccase participates in various physiological functions in lepidopteran insects. As a cell surface protein, laccase enables pathogens to evade immune recognition, playing a multifunctional role in evading insect immune defenses. Furthermore, laccase is crucial for the construction and tanning of the cuticle during pupation and eclosion in insects. This invention contains a laccase inhibitor in a compound of symbiotic bacterial fermentation products from insect pathogenic nematodes to inhibit laccase in tomato leafminer larvae. After laccase inhibition, the tomato leafminer larvae's cuticle becomes lighter, their cuticle softens and tanned, their body color darkens, and their immunity is suppressed while their growth and development are hindered.

[0039] The fermentation broth containing symbiotic bacteria of entomopathogenic nematodes contains toxic proteins that affect tomato leafminer larvae. These larvae exhibit stomach poisoning effects when feeding on tomato leaves containing the fermentation products of the symbiotic bacteria. The green tangerine oil of this invention is a pure natural plant essential oil, extracted from green tangerines using a low-temperature distillation process. Its main component is D-limonene. Green tangerine oil not only facilitates the rapid absorption of the fermentation broth compound by plant leaves but also enhances the adhesion of the liquid to plant leaves, effectively preventing rainwater erosion and improving the utilization rate of fertilizers and pesticides. It also possesses foliar insecticidal and bactericidal effects, is natural with no residue, and has good safety.

[0040] Lycopene, molecular formula C 40 H 56 It is a low-toxicity, high-activity polyphenol oxidase inhibitor extracted from tomatoes. Through the attraction of feed, mature tomato leafminer larvae actively consume lycopene, effectively suppressing the larvae's humoral immune function. This allows entomopathogenic nematodes to more quickly and easily infect and kill mature larvae and cocoons, preventing them from emerging as adults and reducing the adult insect population density in the greenhouse.

[0041] 1-Nonol and ethyl octanoate have significant oviposition attraction activity for tomato leafminer. Female tomato leafminers waiting to lay eggs are attracted to simulated tomatoes (with leaves) to lay eggs, get stuck and die, reducing the chance of female tomato leafminers laying eggs on leaves and fruits, and reducing the overall number of eggs laid in the greenhouse (since the females are dead, they can no longer lay eggs), thus reducing the larval population density.

[0042] Compared with the prior art, the present invention has the following beneficial effects:

[0043] This invention is scientifically designed and ingeniously conceived, consisting of three parts: a compound fermentation product of symbiotic bacteria containing entomopathogenic nematodes, a compound preparation of entomopathogenic nematodes, and a simulated tomato leaf-bearing moth-attracting ball. The compound fermentation product is sprayed onto tomato leaves, while the compound preparation of entomopathogenic nematodes is evenly dripped onto the roots of tomato seedlings during transplanting. This inhibits the humoral immunity of tomato leafminer larvae, making it easier and faster for the entomopathogenic nematodes to infect and kill mature larvae and cocoons, preventing them from emerging as adults and reducing the adult moth population density in the greenhouse. The simulated tomato leaf-bearing moth-attracting ball reduces the opportunity for female tomato leafminers to lay eggs on leaves and fruits, thus reducing the overall egg-laying amount in the greenhouse and lowering the larval population density. This invention achieves its control of tomato leafminers through the synergistic effect of multiple components. Attached Figure Description

[0044] Appendix Figure 1 This is a photograph of the simulated leafy tomato moth-attracting ball of the present invention.

[0045] Appendix Figure 2 This is a layout diagram of the experimental plots and survey arrangements in a tomato greenhouse. Detailed Implementation

[0046] The present invention will be further described in detail below with reference to specific embodiments. These descriptions are for illustrative purposes only and not for limiting the scope of the invention. Unless otherwise specified, specific conditions in the embodiments are performed under conventional conditions or conditions recommended by the manufacturer. Reagents or instruments used, unless otherwise specified, are all commercially available products.

[0047] The glutinous rice glue described in this embodiment of the invention is food-grade glutinous rice glue.

[0048] The entomopathogenic nematode described in this invention is a commercially available strain that is resistant to high temperatures. The entomopathogenic nematode used in the embodiments of this invention is *Steinernema carpocapsae*.

[0049] The LB culture medium described in this invention is existing technology. The LB culture medium in this embodiment of the invention comprises: 10 g / L tryptone; 5 g / L yeast extract; and 10 g / L sodium chloride.

[0050] Example 1

[0051] This embodiment discloses a method for preparing an insect pathogen composition for controlling tomato leafminer according to the present invention. The insect pathogen composition includes a compound of symbiotic bacterial fermentation products of insect pathogenic nematodes sprayed on tomato leaves, a compound preparation of insect pathogenic nematodes applied in holes during tomato seedling transplanting, and a simulated tomato leaf-bearing moth-attracting ball for trapping female tomato leafminers awaiting mating.

[0052] 1. The formulation of the compound agent of symbiotic fermentation products of *Protozoa* is shown in the table below:

[0053] Table 1. Formulation of the compound agent containing symbiotic fermentation products of entomopathogenic nematodes in Example 1.

[0054] composition content Fermentation broth of symbiotic bacteria of entomopathogenic nematodes 200ml / L Plant repellent for tomato leafminer: dried rosemary plant powder 50g / L Laccase inhibitor: 2,3-dimercaptopropanol 5g / L Moisturizer - Trehalose 1g / L Ethoxy-modified polytrisiloxane 3g / L Green orange peel oil 3ml / L water Fill the volume to 1L

[0055] The preparation method of the fermentation product compound of *Protozoa* is as follows:

[0056] S1. Preparation of fermentation broth from symbiotic bacteria of entomopathogenic nematodes:

[0057] Newly acquired infective nematodes were disinfected by soaking them three times in a 0.1% formaldehyde solution for 30 minutes each time, with washing with double-distilled water during the process. Subsequently, the nematodes were soaked in a 1 g / L sodium thiosalicylate solution for another 30 minutes, followed by washing three times with double-distilled water. The nematodes were then ground, and a small amount of the suspension was pipetted onto NBTA plates and incubated at 28°C in the dark for 5 days. Blue-green single colonies were picked and cultured on new NBTA plates, and the process was repeated three times. After purification, single colonies were picked and cultured on LB medium at 28°C and 200 rpm for 15 hours to prepare a fermentation seed culture. This seed culture was inoculated at a rate of 2% into Erlenmeyer flasks containing LB liquid medium, and fermented under the above conditions for 48 hours. The supernatant was collected by high-speed centrifugation; this fermentation supernatant is the fermentation broth of the symbiotic bacteria of the entomopathogenic nematode.

[0058] S2. Preparation of plant repellent: The rosemary plant was dried and ground into powder (80 mesh) to obtain rosemary powder, which was used as a plant repellent.

[0059] S3. Prepare all raw materials according to the formula, mix all raw materials except water evenly, and then add water to make up to the specified volume to obtain the compound of symbiotic bacterial fermentation products of entomopathogenic nematodes.

[0060] 2. The formulation of the compound preparation of entomopathogenic nematodes is shown in the table below:

[0061] Table 2. Formulation of the compound preparation of entomopathogenic nematodes in Example 1

[0062]

[0063] The preparation method of compound preparations of entomopathogenic nematodes is as follows:

[0064] Step (1): Preparation of entomopathogenic nematodes infected and propagated by 5-6 instar larvae of the wax moth: The 5-6 instar larvae of the wax moth were infected and propagated; the 3rd instar larvae (IJs) of the entomopathogenic nematodes were collected in distilled water using the White trap method and stored in a refrigerator at 4°C for later use.

[0065] Step (2): Preparation of insect attractant: Mix the components of the insect attractant evenly according to the specified ratio, and set aside for later use.

[0066] Step (3): Mix the entomopathogenic nematodes that have been infected and propagated by the 5th to 6th instar larvae of the large wax moth, the pest attractant, lycopene, and polyacrylamide in a certain proportion, add water to make up to the specified volume, and obtain a gel-like compound preparation of entomopathogenic nematodes.

[0067] 3. Simulated tomato moth-attracting ball with leaves

[0068] As attached Figure 1 As shown, the simulated tomato moth-attracting ball includes a simulated tomato with leaves and a sponge rod impregnated with 1-nonanol and ethyl octanoate placed on it. The surface of the simulated tomato with leaves and the leaf surface are coated with strong adhesive.

[0069] The preparation method of sponge rods impregnated with 1-nonanol and ethyl octanoate is as follows: Immerse the sponge rods in ethyl octanoate or 1-nonanol or a mixture of ethyl octanoate and 1-nonanol (weight ratio 100:20), remove them, and let them air dry at room temperature until no liquid drips.

[0070] Then, a sponge stick soaked in 1-nonanol and ethyl octanoate is inserted above the simulated tomato with leaves, and then the simulated tomato with leaves and the surface of the leaves are coated with strong glue.

[0071] When using the insect pathogen composition of this embodiment, spray the compound agent of symbiotic bacterial fermentation product of insect pathogenic nematodes onto tomato leaves, applying 15-20L per acre; when transplanting tomato seedlings, evenly drip the compound agent of insect pathogenic nematodes into a circle with a diameter of 10-15cm, centered on the base of the seedling root, at a drip rate of 20-25ml per seedling; hang simulated leafy tomato moth-attracting balls in the tomato field at a height of 20-40cm from the ground, with 20-30 balls hung per acre.

[0072] Example 2

[0073] This embodiment discloses a method for preparing an insect pathogen composition for controlling tomato leafminer according to the present invention. The insect pathogen composition includes a compound of symbiotic bacterial fermentation products of insect pathogenic nematodes sprayed on tomato leaves, a compound preparation of insect pathogenic nematodes applied in holes during tomato seedling transplanting, and a simulated tomato leaf-bearing moth-attracting ball for trapping female tomato leafminers awaiting mating.

[0074] 1. The formulation of the compound agent of symbiotic fermentation products of *Protozoa* is shown in the table below:

[0075] Table 3. Formulation of the compound agent of symbiotic fermentation product of entomopathogenic nematodes in Example 2.

[0076] composition content Fermentation broth of symbiotic bacteria of entomopathogenic nematodes 300ml / L Plant repellent for tomato leafminer: dried rosemary plant powder 25g / L Laccase inhibitor: 2,3-dimercaptopropanol 4g / L Moisturizer - Trehalose 0.5g / L Ethoxy-modified polytrisiloxane 5g / L Green orange peel oil 4ml / L water Fill the volume to 1L

[0077] The preparation method of the fermentation product compound of *Protozoa* is as follows:

[0078] S1. Preparation of fermentation broth from entomopathogenic nematodes:

[0079] Newly acquired infective nematodes were disinfected by soaking them three times in a 0.1% formaldehyde solution for 30 minutes each time, with washing with double-distilled water during the process. Subsequently, the nematodes were soaked in a 1 g / L sodium thiosalicylate solution for another 30 minutes, followed by washing three times with double-distilled water. The nematodes were then ground, and a small amount of the suspension was pipetted onto NBTA plates and incubated at 28°C in the dark for 3 days. Blue-green single colonies were picked and transferred to new NBTA plates for further incubation, repeating the purification process twice. After purification, single colonies were picked and transferred to LB medium, incubated at 28°C and 100 rpm for 24 hours to prepare a fermentation seed culture. This seed culture was inoculated at a 4% inoculum into Erlenmeyer flasks containing LB liquid medium and fermented under the above conditions for 24 hours. The supernatant was collected by high-speed centrifugation; this fermentation supernatant is the fermentation broth of the symbiotic bacteria of the entomopathogenic nematode.

[0080] S2. Same as Example 1.

[0081] S3. Same as Example 1.

[0082] 2. The formulation of the compound preparation of entomopathogenic nematodes is shown in the table below:

[0083] Table 4. Formulation of the compound preparation of entomopathogenic nematodes in Example 2

[0084]

[0085] The preparation method of compound preparations of entomopathogenic nematodes is as follows:

[0086] Step (1): Preparation of entomopathogenic nematodes infected and propagated by 5-6 instar larvae of the wax moth: The 5-6 instar larvae of the wax moth were infected and propagated; the 3rd instar larvae (IJs) of the entomopathogenic nematodes were collected in distilled water using the White trap method and stored in a refrigerator at 4°C for later use.

[0087] Step (2): Mix the entomopathogenic nematodes that have been infected and propagated by the 5th to 6th instar larvae of the large wax moth, the components of the insect attractant, lycopene, and polyacrylamide directly in proportion, add water to make up to the specified volume, and obtain a hydrogel-like compound preparation of entomopathogenic nematodes.

[0088] 3. Simulated tomato moth-attracting ball with leaves

[0089] As attached Figure 1 As shown, the simulated tomato moth-attracting ball includes a simulated tomato with leaves and a sponge rod impregnated with 1-nonanol and ethyl octanoate placed on it. The surface of the simulated tomato with leaves and the leaf surface are coated with strong adhesive.

[0090] The preparation method of sponge rods impregnated with 1-nonanol and ethyl octanoate is as follows: Immerse the sponge rods in ethyl octanoate or 1-nonanol or a mixture of ethyl octanoate and 1-nonanol (weight ratio 100:10), remove them, and let them air dry at room temperature until no liquid drips.

[0091] Then, a sponge stick soaked in 1-nonanol and ethyl octanoate is inserted above the simulated tomato with leaves, and then the simulated tomato with leaves and the surface of the leaves are coated with strong glue.

[0092] When using the insect pathogen composition of this embodiment, spray the compound agent of symbiotic bacterial fermentation product of insect pathogenic nematodes onto tomato leaves, applying 15-20L per acre; when transplanting tomato seedlings, evenly drip the compound agent of insect pathogenic nematodes into a circle with a diameter of 10-15cm, centered on the base of the seedling root, at a drip rate of 20-25ml per seedling; hang simulated leafy tomato moth-attracting balls in the tomato field at a height of 20-40cm from the ground, with 20-30 balls hung per acre.

[0093] Example 3

[0094] This embodiment discloses a method for preparing an insect pathogen composition for controlling tomato leafminer according to the present invention. The insect pathogen composition includes a compound of symbiotic bacterial fermentation products of insect pathogenic nematodes sprayed on tomato leaves, a compound preparation of insect pathogenic nematodes applied in holes during tomato seedling transplanting, and a simulated tomato leaf-bearing moth-attracting ball for trapping female tomato leafminers awaiting mating.

[0095] 1. The formulation of the compound agent of symbiotic fermentation products of *Protozoa* is shown in the table below:

[0096] Table 5. Formulation of the compound agent of symbiotic fermentation product of entomopathogenic nematodes in Example 3.

[0097]

[0098]

[0099] The preparation method of the fermentation product compound of *Protozoa* is as follows:

[0100] S1. Preparation of fermentation broth from entomopathogenic nematodes:

[0101] Newly acquired infective nematodes were disinfected by soaking in a 0.1% formaldehyde solution three times for 30 minutes each time, with washing with double-distilled water during the process. Subsequently, the nematodes were soaked in a 1 g / L sodium thiosalicylate solution for another 30 minutes, followed by washing with double-distilled water three times. The nematodes were then ground, and a small amount of the suspension was pipetted onto NBTA plates and incubated at 28°C in the dark for 5 days. Blue-green single colonies were picked and cultured on new NBTA plates, and the process was repeated three times. After purification, single colonies were picked and cultured on LB medium at 28°C and 400 rpm for 8 hours to prepare a fermentation seed culture. This seed culture was inoculated at a 1% inoculum into Erlenmeyer flasks containing LB liquid medium and fermented under the above conditions for 96 hours. The supernatant was collected by high-speed centrifugation; this fermentation supernatant is the fermentation broth of the entomopathogenic nematode.

[0102] S2. Same as Example 1.

[0103] S3. Same as Example 1.

[0104] 2. The formulation of the compound preparation of entomopathogenic nematodes is shown in the table below:

[0105] Table 6. Formulation of the compound preparation of entomopathogenic nematodes in Example 3.

[0106]

[0107]

[0108] The preparation method of the compound preparation of entomopathogenic nematodes is the same as in Example 1.

[0109] 3. Simulated tomato moth-attracting ball with leaves

[0110] As attached Figure 1 As shown, the simulated tomato moth-attracting ball includes a simulated tomato with leaves and a sponge rod impregnated with 1-nonanol and ethyl octanoate placed on it. The surface of the simulated tomato with leaves and the leaf surface are coated with strong adhesive.

[0111] The preparation method of sponge rods impregnated with 1-nonanol and ethyl octanoate is as follows: Immerse the sponge rods in ethyl octanoate or 1-nonanol or a mixture of ethyl octanoate and 1-nonanol (weight ratio 100:40), remove them, and let them air dry at room temperature until no liquid drips.

[0112] Then, a sponge stick soaked in 1-nonanol and ethyl octanoate is inserted above the simulated tomato with leaves, and then the simulated tomato with leaves and the surface of the leaves are coated with strong glue.

[0113] When using the insect pathogen composition of this embodiment, spray the compound agent of symbiotic bacterial fermentation product of insect pathogenic nematodes onto tomato leaves, applying 15-20L per acre; when transplanting tomato seedlings, evenly drip the compound agent of insect pathogenic nematodes into a circle with a diameter of 10-15cm, centered on the base of the seedling root, at a drip rate of 20-25ml per seedling; hang simulated leafy tomato moth-attracting balls in the tomato field at a height of 20-40cm from the ground, with 20-30 balls hung per acre.

[0114] Example 4

[0115] This embodiment discloses a method for preparing an insect pathogen composition for controlling tomato leafminer according to the present invention. The insect pathogen composition includes a compound of symbiotic bacterial fermentation products of insect pathogenic nematodes sprayed on tomato leaves, a compound preparation of insect pathogenic nematodes applied in holes during tomato seedling transplanting, and a simulated tomato leaf-bearing moth-attracting ball for trapping female tomato leafminers awaiting mating.

[0116] 1. The formulation of the compound agent of symbiotic fermentation products of *Protozoa* is shown in the table below:

[0117] Table 7. Formulation of the compound agent of symbiotic fermentation product of entomopathogenic nematodes in Example 4.

[0118]

[0119]

[0120] The preparation method of the compound agent of symbiotic bacterial fermentation product of the protoneworm is the same as that in Example 1.

[0121] 2. The formulation of the compound preparation of entomopathogenic nematodes is shown in the table below:

[0122] Table 8. Formulation of the compound preparation of entomopathogenic nematodes in Example 4

[0123]

[0124] The preparation method of the compound preparation of entomopathogenic nematodes is the same as in Example 1.

[0125] 3. Simulated tomato moth-attracting ball with leaves

[0126] As attached Figure 1 As shown, the simulated tomato moth-attracting ball includes a simulated tomato with leaves and a sponge rod impregnated with 1-nonanol and ethyl octanoate placed on it. The surface of the simulated tomato with leaves and the leaf surface are coated with strong adhesive.

[0127] The preparation method of sponge rods impregnated with 1-nonanol and ethyl octanoate is as follows: Immerse the sponge rods in ethyl octanoate or 1-nonanol or a mixture of ethyl octanoate and 1-nonanol (weight ratio 100:30), remove them, and let them air dry at room temperature until no liquid drips.

[0128] Then, a sponge stick soaked in 1-nonanol and ethyl octanoate is inserted above the simulated tomato with leaves, and then the simulated tomato with leaves and the surface of the leaves are coated with strong glue.

[0129] When using the insect pathogen composition of this embodiment, spray the compound agent of symbiotic bacterial fermentation product of insect pathogenic nematodes onto tomato leaves, applying 15-20L per acre; when transplanting tomato seedlings, evenly drip the compound agent of insect pathogenic nematodes into a circle with a diameter of 10-15cm, centered on the base of the seedling root, at a drip rate of 20-25ml per seedling; hang simulated leafy tomato moth-attracting balls in the tomato field at a height of 20-40cm from the ground, with 20-30 balls hung per acre.

[0130] Example 5

[0131] This embodiment discloses a method for preparing an insect pathogen composition for controlling tomato leafminer according to the present invention. The insect pathogen composition includes a compound of symbiotic bacterial fermentation products of insect pathogenic nematodes sprayed on tomato leaves, a compound preparation of insect pathogenic nematodes applied in holes during tomato seedling transplanting, and a simulated tomato leaf-bearing moth-attracting ball for trapping female tomato leafminers awaiting mating.

[0132] 1. The formulation of the compound agent of symbiotic fermentation products of *Protozoa* is shown in the table below:

[0133] Table 9. Formulation of the compound agent of symbiotic fermentation product of entomopathogenic nematodes in Example 5.

[0134] composition content Fermentation broth of symbiotic bacteria of entomopathogenic nematodes 250ml / L Plant repellent for tomato leafminer: dried rosemary plant powder 40g / L Laccase inhibitor: 2,3-dimercaptopropanol 3g / L Moisturizer - Trehalose 0.8g / L Ethoxy-modified polytrisiloxane 4g / L Green orange peel oil 1.5ml / L water Fill the volume to 1L

[0135] The preparation method of the compound agent of symbiotic bacterial fermentation product of the protoneworm is the same as that in Example 1.

[0136] 2. The formulation of the compound preparation of entomopathogenic nematodes is shown in the table below:

[0137] Table 10. Formulation of the compound preparation of entomopathogenic nematodes in Example 5

[0138]

[0139]

[0140] The preparation method of the compound preparation of entomopathogenic nematodes is the same as in Example 1.

[0141] 3. Simulated tomato moth-attracting ball with leaves

[0142] As attached Figure 1 As shown, the simulated tomato moth-attracting ball includes a simulated tomato with leaves and a sponge rod impregnated with 1-nonanol and ethyl octanoate placed on it. The surface of the simulated tomato with leaves and the leaf surface are coated with strong adhesive.

[0143] The preparation method of sponge rods impregnated with 1-nonanol is as follows: Immerse the sponge rods in ethyl octanoate, remove them, and let them air dry at room temperature until no liquid drips.

[0144] Then, insert a sponge stick soaked in 1-nonanol into the top of the simulated tomato with leaves, and then coat the simulated tomato with leaves and the surface of the leaves with strong glue.

[0145] When using the insect pathogen composition of this embodiment, spray the compound agent of symbiotic bacterial fermentation product of insect pathogenic nematodes onto tomato leaves, applying 15-20L per acre; when transplanting tomato seedlings, evenly drip the compound agent of insect pathogenic nematodes into a circle with a diameter of 10-15cm, centered on the base of the seedling root, at a drip rate of 20-25ml per seedling; hang simulated leafy tomato moth-attracting balls in the tomato field at a height of 20-40cm from the ground, with 20-30 balls hung per acre.

[0146] Example 6

[0147] The embodiments disclose a method for preparing the insect pathogen composition for controlling tomato leafminer of the present invention. The insect pathogen composition includes a compound of symbiotic bacterial fermentation products of insect pathogenic nematodes sprayed on tomato leaves, a compound preparation of insect pathogenic nematodes applied in holes during tomato seedling transplanting, and a simulated tomato leaf-bearing moth-attracting ball for trapping female tomato leafminers in preparation for mating.

[0148] 1. The formulation of the compound agent of symbiotic fermentation products of *Protozoa* is shown in the table below:

[0149] Table 11 Formulation of the symbiotic fermentation product compound of entomopathogenic nematodes in Example 6

[0150] composition content Fermentation broth of symbiotic bacteria of entomopathogenic nematodes 150ml / L Plant repellent for tomato leafminer: dried rosemary plant powder 65g / L Laccase inhibitor: 2,3-dimercaptopropanol 8g / L Moisturizer - Trehalose 2g / L Ethoxy-modified polytrisiloxane 2.5 / L Green orange peel oil 5ml / L water Fill the volume to 1L

[0151] The preparation method of the compound agent of symbiotic bacterial fermentation product of the protoneworm is the same as that in Example 1.

[0152] 2. The formulation of the compound preparation of entomopathogenic nematodes is shown in the table below:

[0153] Table 12. Formulation of the compound preparation of entomopathogenic nematodes in Example 6

[0154]

[0155]

[0156] The preparation method of the compound preparation of entomopathogenic nematodes is the same as in Example 1.

[0157] 3. Simulated tomato moth-attracting ball with leaves

[0158] As attached Figure 1As shown, the simulated tomato moth-attracting ball includes a simulated tomato with leaves and a sponge rod impregnated with 1-nonanol and ethyl octanoate placed on it. The surface of the simulated tomato with leaves and the leaf surface are coated with strong adhesive.

[0159] The preparation method of sponge rods impregnated with 1-nonanol and ethyl octanoate is as follows: Immerse the sponge rods in ethyl octanoate or 1-nonanol or a mixture of ethyl octanoate and 1-nonanol (weight ratio 100:40), remove them, and let them air dry at room temperature until no liquid drips.

[0160] Then, a sponge stick soaked in 1-nonanol and ethyl octanoate is inserted above the simulated tomato with leaves, and then the simulated tomato with leaves and the surface of the leaves are coated with strong glue.

[0161] When using the insect pathogen composition of this embodiment, spray the compound agent of symbiotic bacterial fermentation product of insect pathogenic nematodes onto tomato leaves, applying 15-20L per acre; when transplanting tomato seedlings, evenly drip the compound agent of insect pathogenic nematodes into a circle with a diameter of 10-15cm, centered on the base of the seedling root, at a drip rate of 20-25ml per seedling; hang simulated leafy tomato moth-attracting balls in the tomato field at a height of 20-40cm from the ground, with 20-30 balls hung per acre.

[0162] Comparative Example 1

[0163] Compared with Example 1, this comparative example does not contain the fermentation product compound of entomopathogenic nematodes, and all other conditions are the same.

[0164] Comparative Example 2

[0165] Compared with Example 1, the comparative example did not contain the laccase inhibitor 2,3-dimercaptopropanol in the symbiotic fermentation product compound of entomopathogenic nematodes, while all other conditions were the same.

[0166] Comparative Example 3

[0167] Compared with Example 1, the fermentation product compound of entomopathogenic nematodes in this comparative example does not contain dried rosemary plant powder, while all other conditions are the same.

[0168] Comparative Example 4

[0169] Compared with Example 1, this comparative example does not contain the compound preparation of entomopathogenic nematodes, and all other conditions are the same.

[0170] Comparative Example 5

[0171] Compared with Example 1, this comparative example did not have a sponge stick soaked in 1-nonanol and ethyl octanoate placed on the simulated tomato moth-attracting ball with leaves, while all other conditions were the same.

[0172] Experimental Example 1

[0173] This experiment investigated the effects of the insect pathogen compositions of Example 1, Comparative Examples 1-5, and commercially available formulations on the control of tomato leafminer. Details are as follows:

[0174] 1. Trial grouping and drugs

[0175] Table 13 Trial Groups and Drugs

[0176] Grouping drug Process 1 group Insect pathogen composition of Example 1 2 groups were treated Comparative Example 1 Insect Pathogen Composition 3 groups were treated Comparative Example 2 Insect Pathogen Composition 4 groups were treated Comparative Example 3 Insect Pathogen Composition 5 groups were treated Comparative Example 4 Insect Pathogen Composition 6 groups were treated Comparative Example 5 Insect Pathogen Composition Blank control group No medication Conventional treatment control group Chlorantraniliprole suspension

[0177] 2. Test Methods

[0178] 2.1 Test Site

[0179] The field trial was conducted at a tomato planting base in Qingliu Town, Xindu District, Chengdu. The greenhouse at the base was a north-facing structure, 40m long and 12m wide. Three greenhouses with a history of severe tomato leafminer infestation were selected for the trial. The tomato seedlings inside were transplanted in early March. Conventional pesticide treatment: 200g·L⁻¹ -1 Chlorantraniliprole suspension, at a dosage of 20 ml / acre (FMC Corporation, USA), was applied using a Delixi 16L electric backpack sprayer, with a water consumption of 45 L per acre; blank control group: no pesticides were applied.

[0180] 2.2 Test Methods

[0181] The experiment consisted of three greenhouses, serving as three replicates. Each greenhouse was divided into eight experimental plots, spaced 2 meters apart and 1 meter from the greenhouse edge. Each experimental plot was 8 meters long and 4 meters wide, with treatment plots arranged in a randomized block design within the greenhouse. The layout and survey arrangement of the experimental plots in the tomato greenhouse are shown in the diagram below. Figure 2 As shown.

[0182] During the tomato harvest season in July, a "W"-shaped even selection of sites was adopted in each plot. Five survey points were selected in each area, and five tomato plants were randomly surveyed at each point. The number of plants damaged by tomato leafminer and the number of damaged fruits were recorded. Two leaves were randomly selected from the upper, middle and lower parts of each tomato plant, and the number of damaged leaves was recorded.

[0183] 2.3 Data Statistics and Analysis

[0184] After the survey data were processed using Excel 2019, SPSS 25.0 was used to perform one-way ANOVA (LSD test) to compare and test the differences between treatments.

[0185] 3. Statistics and Results

[0186] Table 14 Results of different formulations in controlling tomato leafminer

[0187] Number of damaged plants Number of damaged fruits Number of damaged leaves Process 1 group 5.0±1.0e 22.3±2.5e 33.3±3.2d 2 groups were treated 9.7±1.5bc 32.7±2.1cd 43.3±4.0cd 3 groups were treated 6.3±1.5de 43.7±2.1b 37.3±4.0cd 4 groups were treated 8.0±2.0cd 36.7±4.7bcd 39.3±3.1cd 5 groups were treated 11.0±1.0b 31.0±3.6d 46.3±4.7bc 6 groups were treated 11.0±1.0b 41.7±3.1bc 54.7±6.5b Blank control group 13.7±1.5a 64.3±9.3a 77.3±9.7a Conventional treatment control group 4.0±1.0e 33.7±6.7cd 23.0±5.6e

[0188] It can be seen from the above table:

[0189] (1) In terms of the number of plants damaged, treatment group 1 and the conventional control group showed the best performance, with no significant difference between them, but were significantly better than treatment group 2 (containing the symbiotic fermentation product compound of entomopathogenic nematodes), treatment group 5 (containing the compound preparation of entomopathogenic nematodes), and treatment group 6 (simulated tomato moth-attracting balls without sponges soaked in 1-nonanol and ethyl octanoate). The blank control group had the highest number of damaged plants, but was significantly worse than all treatment groups. This indicates that the symbiotic fermentation product compound of entomopathogenic nematodes, the compound preparation of entomopathogenic nematodes, and the sponges soaked in 1-nonanol and ethyl octanoate all play a crucial and indispensable role in controlling the damage of tomato leafminer to tomato plants.

[0190] (2) In terms of the number of damaged fruits, treatment group 1 showed the best control effect, which was significantly better than the conventional control group and other treatment groups. The number of damaged fruits in treatment group 6 (the simulated tomato moth bait ball without sponge sticks soaked in 1-nonanol and ethyl octanoate) was significantly higher than that in treatment group 1 and treatment group 5, indicating that the sponge sticks soaked in 1-nonanol and ethyl octanoate played an extremely important and indispensable role in fruit protection.

[0191] (3) In terms of the number of damaged leaves, the conventional control group performed best, followed by treatment group 1. The blank control group had the highest number of damaged leaves, followed by treatment groups 6 and 5. This indicates that the compound preparation of entomopathogenic nematodes and the sponge sticks impregnated with 1-nonanol and ethyl octanoate both played a crucial and indispensable role in controlling the damage of tomato leafminer to tomato leaves.

[0192] 4. Conclusion

[0193] Treatment group 1, which uses the insect pathogen composition of the present invention for controlling tomato leafminer, showed the best performance in terms of the number of damaged plants and fruits, and had the best overall control effect.

[0194] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A combination product for controlling the insect pathogen of tomato leafminer, characterized in that, This includes a compound of symbiotic bacteria fermentation products of entomopathogenic nematodes sprayed on tomato leaves, a compound of entomopathogenic nematodes applied in holes during tomato seedling transplanting, and a simulated tomato moth-attracting ball with leaves for trapping female tomato leafminers awaiting birth. Each 1L of the compound preparation of symbiotic fermentation products of entomopathogenic nematodes contains 100-400ml of entomopathogenic nematode symbiotic fermentation broth, 25-100g of tomato leafminer plant repellent, 2.5-10g of laccase inhibitor, 0.5-2g of humectant, 1.5-6g of ethoxylated modified polytrisiloxane, 1.5-6ml of green orange peel oil, and water to make up the volume. Each 1L of the compound preparation of entomopathogenic nematodes contains 5,000,000 to 20,000,000 IJs of entomopathogenic nematodes that have been infected and propagated by 5-6 instar larvae of the large wax moth, 0.5 to 2 mol of polyphenol oxidase inhibitor, 2.5 to 10 g of nematode moisturizer, and 50 to 300 g of pest attractant, with water added to make up the volume; The simulated tomato moth-attracting ball includes a simulated tomato with leaves and a sponge stick impregnated with 1-nonanol and / or ethyl octanoate placed on it, the surface of which is coated with a strong adhesive. The entomopathogenic nematodes are of the genus Strychnine. Steinernema or / and Heterostachys genus Heterorhabditis Nematodes.

2. The insect pathogen combination product for controlling tomato leafminer according to claim 1, characterized in that, Each 1L of the compound of symbiotic fermentation products of entomopathogenic nematodes contains 200ml of entomopathogenic nematode symbiotic fermentation broth, 50g of tomato leafminer plant repellent, 5g of laccase inhibitor, 1g of humectant, 3g of ethoxylated polytrisiloxane, 3ml of green orange peel oil, and water to make up the volume. Or / and each 1L of the compound preparation of entomopathogenic nematodes contains 10,000,000 IJs of entomopathogenic nematodes that have been infected and propagated by 5-6 instar larvae of the large wax moth, 1mol of polyphenol oxidase inhibitor, 5g of nematode moisturizer, and 65.5-262g of pest attractant, with water to make up the volume.

3. The insect pathogen combination product for controlling tomato leafminer according to claim 2, characterized in that, Each 1L of the compound preparation of entomopathogenic nematodes contains 131g of insect attractant.

4. The insect pathogen combination product for controlling tomato leafminer according to claim 2, characterized in that, Each 1L of the compound preparation for entomopathogenic nematodes contains the following insect attractants: 10-40g tomato sauce, 15-60g casein, 10-40g wheat germ powder, 5-20g soybean flour, 2.5-10g sucrose, 4-16g glucose, 2.5-10g linoleic acid, 1.5-6g soybean oil, 5-20g vitamin C, and 10-40g cellulose.

5. The insect pathogen combination product for controlling tomato leafminer according to claim 2, characterized in that, Each 1L of the compound preparation of entomopathogenic nematodes contains the following insect attractants: 20g tomato sauce, 30g casein, 20g wheat germ powder, 10g soybean powder, 5g sucrose, 8g glucose, 5g linoleic acid, 3g soybean oil, 10g vitamin C, and 20g cellulose.

6. A combination product for controlling tomato leafminer according to any one of claims 1 to 5, characterized in that, The preparation method of the symbiotic fermentation broth of the entomopathogenic nematode includes the following steps: After disinfecting and grinding the entomopathogenic nematodes in the infecting stage, a small amount of nematode suspension is taken and transferred to an NBTA plate. The plate is then cultured at 28±0.5℃ in the dark for 3-5 days. Blue-green single colonies are picked and transferred to a new NBTA plate for further culture. The purification process is repeated 2-3 times. After purification, single colonies are picked and transferred to LB medium. The plate is then cultured at 28±0.5℃ and 100-400 rpm for 8-24 hours to prepare the fermentation seed liquid. The seed liquid is inoculated into an Erlenmeyer flask containing LB liquid medium at an inoculation rate of 1-4%. The flask is fermented under the above conditions for 24-96 hours. The supernatant is then collected by high-speed centrifugation. This fermentation supernatant is the symbiotic fermentation broth of the entomopathogenic nematode. Or / and the tomato leafminer plant repellent includes rosemary powder; Or / and the laccase inhibitor includes 2,3-dimercaptopropanol; Or / and the moisturizer includes trehalose.

7. A combination product for controlling tomato leafminer according to any one of claims 1 to 5, characterized in that, The polyphenol oxidase inhibitor includes lycopene; and / or the nematode moisturizer includes polyacrylamide.

8. A combination product for controlling tomato leafminer according to any one of claims 1 to 5, characterized in that, The preparation method of entomopathogenic nematodes that have been infected and propagated by the 5th-6th instar larvae of the wax moth is as follows: infect and propagate the 5th-6th instar larvae of the wax moth; collect the third instar larvae of the entomopathogenic nematodes propagated by the White trap method in distilled water and store them in a refrigerator at 4°C for later use.

9. A method for preparing an insect pathogen combination product for controlling tomato leafminer according to any one of claims 1 to 8, characterized in that, A compound preparation of symbiotic bacterial fermentation products, a compound preparation of insect pathogenic nematodes, and a simulated leafy tomato moth-attracting ball were prepared respectively. The preparation method of the compound agent of symbiotic fermentation product of entomopathogenic nematodes includes the following steps: take the fermentation liquid of entomopathogenic nematodes symbiotic bacteria, tomato leafminer plant repellent, laccase inhibitor, moisturizer, ethoxylated modified polytrisiloxane and green orange oil according to the formula, mix them evenly, and make up the volume with water; The compound preparation of entomopathogenic nematodes includes the following steps: take the entomopathogenic nematodes, polyphenol oxidase inhibitors, nematode moisturizers and insect attractants according to the formula, mix them evenly, and make up the volume with water; or first mix the insect attractant components evenly, and then mix them evenly with the formula amount of entomopathogenic nematodes, polyphenol oxidase inhibitors and nematode moisturizers, and make up the volume with water. The preparation method of the simulated leafy tomato moth-attracting ball includes the following steps: First, immerse the sponge stick in ethyl octanoate or 1-nonanol or a mixture of ethyl octanoate and 1-nonanol, take it out, and let it dry at room temperature until it no longer drips liquid. Then, insert the sponge stick into the top of the simulated leafy tomato.

10. A method for preparing an insect pathogen combination product for controlling tomato leafminer according to claim 9, characterized in that, When the sponge rod is immersed in a mixture of ethyl octanoate and 1-nonanol, the weight ratio of ethyl octanoate to 1-nonanol is 100:10~40.

11. The method for preparing an insect pathogen combination product for controlling tomato leafminer according to claim 9, characterized in that, When the sponge rod is immersed in a mixture of ethyl octanoate and 1-nonanol, the weight ratio of ethyl octanoate to 1-nonanol is 100:

20.

12. The application of the insect pathogen combination product for controlling tomato leafminer according to any one of claims 1 to 8 in the control of tomato leafminer.

13. A method for controlling tomato leafminer, characterized in that, The insect pathogen combination product according to any one of claims 1 to 8 is used.

14. The method according to claim 13, characterized in that, Spray the compound of fermentation products of symbiotic bacteria of entomopathogenic nematodes onto tomato leaves at a rate of 15-20L per acre. When transplanting tomato seedlings, apply a compound preparation of insect pathogenic nematodes evenly to the roots of the tomato seedlings. Suspend simulated tomato moth-attracting balls with leaves in the tomato field.

15. The method according to claim 14, characterized in that, When transplanting tomato seedlings, use the base of the seedling as the center and evenly drip the compound preparation of entomopathogenic nematodes into a circle with a diameter of 10-15cm.

16. The method according to claim 14, characterized in that, When transplanting tomato seedlings, apply a compound preparation of insect pathogenic nematodes evenly to the roots of the tomato seedlings, at a rate of 20-25 ml per seedling.

17. The method according to claim 14, characterized in that, The simulated tomato moth-attracting ball with leaves should be suspended at a height of 20-40cm from the ground.

18. The method according to claim 14, characterized in that, Hang 20-30 simulated tomato moth-attracting balls with leaves on each acre of land.

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