A pesticide synergist and a method for enhancing efficacy

Abstract

This invention relates to the field of agricultural pest control and pesticide reduction with enhanced efficacy, specifically to a pesticide synergist and a method for enhancing efficacy. The active ingredients of the pesticide synergist consist of tannic acid, rutin, and quercetin. The synergistic method involves sequentially applying a chemical pesticide and the pesticide synergist to the plant, with the synergist applied three days after the application of the chemical pesticide. The method also includes a second application seven days after the first application of the synergist, and foliar spraying or root drenching with the synergist at a concentration of 2-5 g / L. Under conditions of a 25% reduction in chemical pesticide dosage, the combined application of 5 g / L of the synergist resulted in control efficacy against various pests, including two-spotted spider mites and aphids, exceeding the effect of using the full-volume pesticide alone, with a control efficacy reaching 88.62% 14 days after application. The mechanism involves activating the plant's defense enzyme system, initiating secondary metabolic pathways, and inducing cell wall thickening; the three active ingredients, when combined, exhibit a significant synergistic effect.

Description

Technical Field

[0001] This invention relates to the field of agricultural pest control and pesticide reduction and efficiency enhancement, specifically to a pesticide synergist and a method for enhancing efficacy. Background Technology

[0002] Chemical pesticides are the primary means of controlling agricultural pests, especially those with wide distribution, rapid reproduction, and quickly developing resistance, such as two-spotted spider mites, aphids, whiteflies, and diamondback moths. However, long-term over-reliance and excessive use of chemical pesticides have not only led to a continuous increase in pest resistance and decreased control effectiveness, but have also resulted in a series of serious problems, including excessive pesticide residues, environmental pollution, and risks to the quality and safety of agricultural products. Therefore, how to significantly reduce the use of chemical pesticides without affecting, or even improving, control effectiveness has become a key technical challenge that urgently needs to be addressed for the green and sustainable development of agriculture.

[0003] Plants possess a complex physical and chemical defense system, capable of resisting pest infestation by producing secondary metabolites (such as tannins and flavonoids) and strengthening structural barriers (such as cell wall thickening). Existing research has confirmed that some plant-derived bioactive substances possess inherent insecticidal activity or the potential to induce insect resistance in plants. However, current technologies typically focus only on the direct insecticidal or insect-suppressive effects of these single substances, or remain at the level of research on their basic physiological activities.

[0004] In practical applications, developing these plant-derived active substances into highly efficient and stable pesticide synergists still faces several bottlenecks: First, how to achieve stable and efficient delivery. These substances vary in water solubility and stability, and simple physical mixing or aqueous solutions cannot guarantee their stability and bioavailability in field application. Second, how to scientifically synergize with existing chemical pesticides. Simple tank mixing may lead to compatibility problems or reduced efficacy, while the timing and sequence of application lack scientific basis, making it difficult to leverage the synergistic effect of mixing. Third, how to maximize the activation of plant systemic resistance. It is not yet clear whether exogenous application of these substances can systematically and persistently activate multiple plant defense pathways (such as defense enzyme systems, secondary metabolic pathways, and physical structure enhancement), thereby forming an integrated control strategy that complements the rapid effects of chemical pesticides.

[0005] Therefore, there is an urgent need in this field for a pesticide synergist and its supporting application technology that can not only work synergistically with chemical pesticides to achieve significant "reduction and efficiency", but also stably and systematically activate the plant's own immune defense, thereby constructing a long-term green control system. Summary of the Invention

[0006] To address the problems existing in the prior art, the present invention provides a pesticide synergist and a method for synergizing pesticide effects.

[0007] This invention is achieved through the following technical solution: A pesticide synergist, the active ingredients of which are composed of tannic acid, rutin and quercetin; the mass percentage of the active ingredients in the pesticide synergist is: 10%-12% tannic acid, 0.5%-1% rutin and 0.5%-1% quercetin.

[0008] Furthermore, its dosage form is a microemulsion.

[0009] Furthermore, the pesticide synergist also includes 10% dimethyl sulfoxide, 10% cyclohexanone, 4% alkylphenol polyoxyethylene ether-10, 4.5% calcium dodecylbenzenesulfonate, 2% polyvinyl alcohol, 0.1% sodium citrate, 1% ethylene glycol, and water to make up to 100%.

[0010] The present invention also provides a method for enhancing the efficacy of pesticides, comprising: sequentially applying a chemical agent and the pesticide enhancer to the plant, wherein the pesticide enhancer is applied on the third day after the application of the chemical agent.

[0011] Furthermore, the pesticide synergist is applied by foliar spraying or root irrigation.

[0012] Furthermore, applying a second synergist 7 days after the first application can enhance the control effect to an even higher level 14 days after application.

[0013] Furthermore, the application concentration of pesticide synergists is 2-5 g / L.

[0014] Furthermore, the chemical agent is used to control one of the following: two-spotted spider mite, diamondback moth, aphid, or whitefly.

[0015] Beneficial technical effects of the present invention: (1) This invention provides a synergist microemulsion with tannic acid, quercetin, and rutin as specific active ingredients, and clarifies the corresponding application techniques. Its core technical effect is to achieve "reduced dosage and increased efficacy" of chemical pesticides, that is, to obtain higher pest control effects while reducing the amount of pesticides used. Specifically, when using 75% of the recommended dosage of chemical pesticides (i.e., a 25% reduction), the application of this synergist at a concentration of 5 g / L significantly improved the control effect on four representative pests on the 7th day after application, compared with the treatment using 100% of the recommended dosage of chemical pesticides alone. Specific data include: the control efficacy against strawberry two-spotted spider mite increased from 52.67% to 75.64%; the control efficacy against cucumber aphid increased from 64.51% to 77.34%; the control efficacy against tomato whitefly increased from 56.67% to 72.26%; and the control efficacy against cabbage diamondback moth increased from 75.91% to 81.69%.

[0016] (2) By optimizing the application procedure ("apply chemical agents first, then apply synergist after 3-5 days", and combine with "apply synergist twice after 7 days"), the duration of the control effect can be significantly extended. For example, against strawberry two-spotted spider mite, the control efficacy can be further increased to 88.62% on the 14th day after application, while the control efficacy of using only full-volume pesticide under the same conditions has dropped to 55.29%.

[0017] (3) The synergist of this invention not only serves as an adjunct to chemical pesticides, but its fundamental role is to act as a "systemic immune activator" for plants, enhancing their inherent insect resistance from multiple biological levels. Specific mechanisms include: ① Rapid activation of the defense enzyme system: After application, it significantly induces the activity of key defense enzymes such as peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) in plants, reaching a peak on the 3rd day after treatment, marking the plant's entry into a highly defensive state. ② Initiation of secondary metabolic defense pathways: It specifically upregulates the expression of key genes in the biosynthesis pathways of flavonoids and lignin, driving the accumulation of insect-resistant secondary metabolites such as flavonoids, phenolic acids, tannins, and lignin, thus constructing a chemical barrier. ③ Enhancement of physical defense structures: By promoting the synthesis of substances such as lignin, it directly induces thickening of plant leaf cell walls, increasing the mechanical resistance to feeding by pests (especially piercing-sucking pests). Meanwhile, the synergistic effect of the three active ingredients is crucial: experiments have shown that when tannic acid, quercetin and rutin are combined in a specific ratio (such as 10:1:1), the repellency rate against two-spotted spider mites is as high as 75.57%, which is significantly better than the repellency rate when any one of the ingredients is used alone. Attached Figure Description

[0018] Figure 1 Changes in the activity of defense enzymes in strawberry plants under synergist treatment; Figure 2 Changes in genes and substances related to flavonoid metabolism pathways in strawberry plants under synergist treatment; Figure 3 Changes in genes and substances related to lignin metabolism pathways in strawberry plants under synergist treatment; Figure 4 The effects of synergist treatment on the ultrastructure of strawberry leaves; Figure 5 The effect of synergist treatment on cell wall thickness; Figure 6 Changes in the activity of defensive enzymes in strawberry plants under two-times and three-times consecutive treatments with synergists; Figure 7 Changes in the expression levels of key genes in the flavonoid pathway in strawberry plants under two-times and three-times consecutive treatments with synergists; Figure 8 The repellency rate of different treatments on two-spotted spider mites; Figure 9 The repulsion rate of strawberry leaves against two-spotted spider mites after tannic acid induction for different number of days; Figure 10 Dynamic changes in tannin spirotetramat content in strawberry leaves TA: Tannic acid; Q: Quercetin; R: Rutin; T1: Tannic acid: Quercetin: Rutin = 12:0.5:0.5; T2: Tannic acid: Quercetin: Rutin = 10:1:1; T3: Tannic acid: Quercetin: Rutin = 5:1:1. Detailed Implementation

[0019] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of this application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0020] The test materials used in the embodiments of this invention are all conventional test materials in the art and can be purchased through commercial channels.

[0021] Experimental materials: Tannic acid was purchased from Tianjin Kaitong Chemical Reagent Co., Ltd. Tested plants: strawberry (Hongyan), cucumber (Zhaibubai), tomato (Yingfen No. 8), and cabbage (Jingfeng No. 1).

[0022] Example 1

[0023] The synergist microemulsion formulation is as follows: 10% tannic acid, 1% rutin, 1% quercetin, 10% dimethyl sulfoxide, 10% cyclohexanone, 4% alkylphenol polyoxyethylene ether-10 (OP-10), 4.5% calcium dodecylbenzenesulfonate (agricultural emulsion 500#), 2% polyvinyl alcohol, 0.1% sodium citrate, 1% ethylene glycol, and water to make up to 100%.

[0024] 1. Differences in the control efficacy of the synergist prepared in Example 1 and pesticides applied in different orders against strawberry two-spotted spider mite, cucumber aphid, tomato whitefly, and cabbage diamondback moth larvae. Preparation of the medicine solution: The enhanced microemulsion was diluted with deionized water to three concentrations: 1 g / L, 2 g / L, and 5 g / L.

[0025] Commercial 30% etoxazole suspension was diluted with deionized water to prepare aqueous solutions of active ingredient at 75 mg / L and 100 mg / L (i.e., 75% and 100% of the recommended dose).

[0026] Dilute commercial 20% acetamiprid soluble solution with deionized water to prepare aqueous solutions of 18.75 mg / L and 25 mg / L of active ingredient (i.e., 75% and 100% of the recommended dose).

[0027] Dilute commercial 25 g / L bifenthrin emulsifiable concentrate with deionized water to prepare aqueous solutions of active ingredients at 18.75 mg / L and 25 mg / L (i.e., 75% and 100% of the recommended dose).

[0028] Dilute commercial 1.8% avermectin emulsion with deionized water to prepare aqueous solutions of 13.5 mg / L and 18 mg / L of active ingredient (i.e., 75% and 100% of the recommended dose).

[0029] Application Implementation: (1) Potted strawberry seedlings with uniform growth and cultured for two weeks were selected for the experiment (one seedling per pot). Different concentrations of synergist (1 g / L, 2 g / L, 5 g / L) were sprayed on each pot at 10 mL. A control group was set up by spraying each pot with 75 mg / L and 100 mg / L aqueous solutions of etoxazole. A water control group was also set up.

[0030] (2) Potted cucumber seedlings with uniform growth and cultured for two weeks were selected for the experiment (one seedling per pot). Different concentrations of synergist (1 g / L, 2 g / L, 5 g / L) were sprayed on each pot at 10 mL. A pesticide control was set up, and acetamiprid 18.75 mg / L and 25 mg / L aqueous solutions were sprayed on each pot at 10 mL. A water control was also set up.

[0031] (3) Potted tomato seedlings with uniform growth and cultured for two weeks were selected for the experiment (one seedling per pot). Different concentrations of synergist (1 g / L, 2 g / L, 5 g / L) were sprayed on each pot at 10 mL. A control group was set up by spraying each pot with 18.75 mg / L and 25 mg / L aqueous solutions of bifenthrin. A water control group was also set up.

[0032] (4) Potted cabbage seedlings with uniform growth and reaching the 5-6 leaf stage were selected for the experiment (one seedling per pot). Different concentrations of synergist (1 g / L, 2 g / L, 5 g / L) were sprayed at 10 mL per pot. A control group was set up by spraying 10 mL of abamectin aqueous solution at 13.5 mg / L and 18 mg / L per pot. A water control group was also set up. The grouping of each treatment in the potted experiment is shown in Table 1.

[0033] Table 1. Pot Experiment Grouping

[0034] Five treatments of seedlings were cultured in an artificial climate chamber under the following conditions: temperature 20±2℃, relative humidity (RH) 70%, and light / dark cycle = 14:10 h. Each treatment was replicated in triplicate, with three plants per replicate, and each replicate was spaced apart. Each strawberry seedling was inoculated with 80 two-spotted spider mites, each cucumber seedling with 50 aphids, each tomato seedling with 80 whiteflies, and each cabbage seedling with 30 diamondback moths. The experiments were conducted three days after the infestation sources stabilized. The number of insects on the leaves of each plant was surveyed one day before application and seven days after application, and the number of surviving insects for each treatment was recorded. The control effect was calculated based on the survey data. The experimental results are shown in Tables 2, 3, 4, and 5.

[0035] Table 2. Control efficacy of three different concentrations and application sequences of synergists on strawberry two-spotted spider mites after 7 days.

[0036] Table 3. Control efficacy of three different concentrations and application sequences of synergists against cucumber aphids after 7 days.

[0037] Table 4. Control effect of three different concentrations and application sequences of synergists on tomato whiteflies after 7 days.

[0038] Table 5. Control efficacy of three different concentrations and application sequences of synergists on diamondback moth of cabbage after 7 days.

[0039] Tables 2, 3, 4, and 5 show that significant differences in control efficacy occurred among the treatment groups 7 days after application. When the synergist concentration was 1 g / L, regardless of the order of application, the control efficacy was generally lower than that of single chemical treatments. When the synergist concentration was 2 g / L, the control efficacy was generally comparable to that of single chemical treatments. When the synergist concentration was 5 g / L, the combination of synergist and reduced-dose chemical treatments resulted in significantly higher control efficacy than single chemical treatments. The treatment of applying 75% of the recommended chemical dose followed by the synergist showed the best control efficacy against strawberry two-spotted spider mite, cucumber aphid, tomato whitefly, and cabbage diamondback moth larvae, significantly better than the effect of using full-dose chemical treatments alone. This combination method is the recommended application method.

[0040] 2. Differences in the control efficacy of synergists against strawberry two-spotted spider mite and cucumber aphids under different application methods (foliar spraying and root drenching). Preparation of the medicine solution: The enhanced microemulsion was diluted with deionized water to three concentrations: 1 g / L, 2 g / L, and 5 g / L.

[0041] Commercial 22.4% spirotetramat suspension was diluted with deionized water to prepare aqueous solutions of 48 mg / L and 64 mg / L.

[0042] Commercial 20% acetamiprid soluble solution was diluted with deionized water to prepare aqueous solutions of 18.75 mg / L and 25 mg / L.

[0043] Application Implementation: (1) Potted strawberry seedlings with uniform growth and cultured for two weeks were selected for the experiment (one plant per pot). Different concentrations of synergist (1 g / L, 2 g / L, 5 g / L) were sprayed at 10 mL per pot and drenched at 10 mL per pot. Spirotetramat at 48 mg / L and 64 mg / L were sprayed at 10 mL per pot, and a water control was set up. In treatment one, 80 two-spotted spider mites were inoculated into each strawberry seedling. After 3 days, when the infestation source was stable, spirotetramat was sprayed. After 6 days, different concentrations of synergist were sprayed at 10 mL per pot. In treatment two, 80 two-spotted spider mites were inoculated into each strawberry seedling. After 3 days, when the infestation source was stable, spirotetramat was sprayed. At the same time, different concentrations of synergist were drenched at 10 mL per pot. In treatment three, 80 two-spotted spider mites were inoculated into each strawberry seedling. After 3 days, when the infestation source was stable, spirotetramat was sprayed. Each treatment had 3 replicates, with 3 plants per replicate, and the replicates were kept at a distance. Each strawberry seedling was inoculated with 80 two-spotted spider mites. The number of test mites on the leaves of each plant was investigated one day before and seven days after application. The number of surviving mites in each treatment was recorded, and the control effect was calculated based on the experimental data.

[0044] (2) Potted cucumber seedlings with uniform growth and cultured for two weeks were selected for the experiment (one seedling per pot). Different concentrations of synergist (1 g / L, 2 g / L, 5 g / L) were sprayed at 10 mL per pot and drenched at 10 mL per pot. Acetamiprid 18.75 mg / L and 25 mg / L aqueous solutions were sprayed at 10 mL per pot, and a water control was set up. In treatment one, 50 aphids were introduced into each cucumber seedling. After 3 days, when the insect source was stable, acetamiprid was sprayed. After 6 days, different concentrations of synergist were sprayed at 10 mL per pot. In treatment two, 50 aphids were introduced into each strawberry seedling. After 3 days, when the insect source was stable, acetamiprid was sprayed. At the same time, different concentrations of synergist were drenched at 10 mL per pot. In treatment three, 50 aphids were introduced into each strawberry seedling. After 3 days, when the insect source was stable, acetamiprid was sprayed. Each treatment had 3 replicates, with 3 plants per replicate, and the replicates were kept at a distance. The number of test insects on the leaves of each plant was investigated one day before and seven days after application. The number of surviving insects in each treatment was recorded. The control effect was calculated based on the experimental data. The experimental results are shown in Tables 6 and 7.

[0045] Table 6. Control efficacy of different application methods of synergists on strawberry two-spotted spider mite 7 days later.

[0046] Table 7. Control efficacy of different application methods of synergists against cucumber aphids 7 days later.

[0047] As shown in Tables 6 and 7, when the concentration of the synergist is 2 g / L-5 g / L, whether it is applied by foliar spraying or root irrigation, its control effect on strawberry two-spotted spider mite and cucumber aphid is higher than that of the chemical agent treatment group alone, and both have a synergistic effect on chemical agents.

[0048] 3. Differences in the control efficacy of synergists at different application rates and intervals against strawberry two-spotted spider mite and cucumber aphid. Preparation of the medicine solution: Dilute the synergistic microemulsion to a concentration of 5 g / L with deionized water.

[0049] Commercial 30% etoxazole suspension was diluted with deionized water to prepare aqueous solutions of 75 mg / L and 100 mg / L.

[0050] Commercial 20% acetamiprid soluble solution was diluted with deionized water to prepare aqueous solutions of 18.75 mg / L and 25 mg / L.

[0051] Application Implementation: (1) Potted strawberry seedlings with uniform growth and cultured for two weeks were selected for the experiment (one seedling per pot). The synergist was diluted to 5 g / L and sprayed with 10 mL per pot; etoxazole was sprayed with 75 mg / L and 100 mg / L at 10 mL per pot. A water control was set up. 80 two-spotted spider mites were inoculated into each strawberry seedling. After 3 days, when the insect source was stable, etoxazole was sprayed, and after 6 days, the synergist was sprayed with 5 g / L at 10 mL per pot. Another treatment group was set up, which was sprayed with 10 mL of synergist again on the 14th day, and the treatment group 2 was sprayed with 10 mL of synergist again on the 10th and 14th days. Each treatment had 3 replicates, with 3 plants in each replicate, and the replicates were kept at a distance. The number of test insects on the leaves of each plant was investigated on the 3rd day before the chemical agent was sprayed and on the 6th day before the synergist was sprayed, as well as on the 7th and 14th days after the application. The number of surviving insects in each treatment was recorded, and the control effect was calculated based on the experimental data.

[0052] (2) Potted cucumber seedlings with uniform growth and cultured for two weeks were selected for the experiment (one seedling per pot). The synergist was diluted to 5 g / L and sprayed with 10 mL per pot; acetamiprid 18.75 mg / L and 25 mg / L aqueous solutions were sprayed with 10 mL per pot. A water control was set up. 50 aphids were introduced into each cucumber seedling. After 3 days, when the insect source was stable, acetamiprid was sprayed, and after 6 days, 5 g / L synergist was sprayed with 10 mL per pot. Another treatment group was set up, which was sprayed with 10 mL of synergist again on the 14th day, and treatment group 2 was sprayed with 10 mL of synergist again on the 10th and 14th days. Each treatment had 3 replicates, with 3 plants in each replicate, and the replicates were kept at a distance. The number of test insects on the leaves of each plant was investigated on the 3rd day before the chemical agent was sprayed and on the 6th day before the synergist was sprayed, as well as on the 7th and 14th days after the application. The number of surviving insects in each treatment was recorded, and the control effect was calculated based on the experimental data. The experimental results are shown in Tables 8 and 9.

[0053] Table 8. Control efficacy of different application times of synergist on strawberry two-spotted spider mite after 7 and 14 days.

[0054] Table 9. Control efficacy of different application methods of synergists against cucumber aphids 7 and 14 days later.

[0055] As shown in Tables 8 and 9, applying the synergist twice at a 7-day interval prolongs the plant's control effect on pests. At 14 days post-application, the control efficacy against strawberry two-spotted spider mites and cucumber aphids reached 88.62% and 85.70%, respectively, significantly higher than the control effect of a single application of the synergist. When the synergist was applied three times at a 3-day interval, the control efficacy at 7 days post-application was higher than that of single and double applications, but the duration of effect was short, and the control efficacy decreased at 14 days post-application, only higher than the effect of using the full dose of chemical agent alone.

[0056] 4. Exploration of the synergistic mechanism of synergists (1) Synergists can activate the activity of plant defense enzymes. Potted strawberry seedlings with uniform growth, cultivated for two weeks, were selected for the experiment (one seedling per pot). The enhanced microemulsion was diluted to 5 g / L with deionized water and sprayed onto the strawberry seedlings at a rate of 10 mL per seedling. Spraying with plain water served as a control. Five strawberry seedlings were treated per group. Samples were collected from both the experimental and control groups at 1, 3, and 7 days after spraying to determine the activity of the strawberry plant's defensive enzymes. Results are as follows: Figure 1As shown, the activities of POD, PPO, and PAL in leaves after application of the synergist generally exhibited a trend of first increasing and then decreasing over time. On the third day after treatment, the activities of POD, PPO, and PAL showed a significant upward trend compared to the control group, reaching their maximum values ​​and being significantly higher than the control group. This indicates that components such as tannic acid, quercetin, and rutin in the synergist can induce an increase in the activity of plant defense enzymes, thus helping to improve the plant's insect resistance.

[0057] (2) Synergists can activate secondary metabolic pathways in plants. Potted strawberry seedlings with uniform growth, cultured for two weeks, were selected for the experiment (one seedling per pot). The enhanced microemulsion was diluted to 5 g / L with deionized water and sprayed onto the strawberry seedlings at a rate of 10 mL per seedling. Spraying with plain water served as a control. Five strawberry seedlings were treated per group. On the third day after spraying, samples were collected from both the experimental and control groups to determine changes in the expression levels of key genes in the flavonoid and lignin metabolic pathways of the strawberry plants, as well as the content of secondary metabolites. Results are as follows: Figure 2 , 3 As shown, the application of the synergist indicates that the tannic acid, quercetin, rutin and other components can activate the flavonoid and lignin metabolic pathways, induce the upregulation of key genes in the secondary metabolic pathways, and accumulate secondary metabolites such as flavonoids, phenolic acids, tannins and lignin, putting the plant in a highly defensive state and enhancing the plant's defense performance.

[0058] (3) Synergists can induce thickening of strawberry leaf cell walls. Potted strawberry seedlings with uniform growth and cultured for two weeks were selected for the experiment (one seedling per pot). A synergistic microemulsion was diluted to 5 g / L with deionized water and sprayed onto the strawberry seedlings at a rate of 10 mL per seedling. Spraying with plain water served as a control. Five strawberry seedlings were treated per group. Samples were taken from both the treatment and control groups on day 5 after spraying. Seven days later, 10 mL of the 5 g / L synergistic agent solution was sprayed onto each seedling again. Samples were taken from both groups again on day 14. Strawberry leaves were cut into 1 mm × 2 mm pieces in 2.5% glutaraldehyde fixative. These pieces, along with the fixative, were drawn into a 5 mL syringe, vacuumed until the leaves settled, and then quickly transferred to 1.5 mL centrifuge tubes. The tubes were labeled, sealed with sealing film, and stored at 4°C for transmission electron microscopy imaging. Results are shown below. Figure 4-5 As shown, the application of the synergist indicates that components such as tannins, quercetin, and rutin, by activating the lignin metabolic pathway, induce an increase in lignin content, promote thickening of strawberry leaf cell walls, increase the feeding resistance of pests, and enhance the plant's physical defense capabilities.

[0059] (4) Effects of repeated application of synergists on the activation of plant defense systems Potted strawberry seedlings with uniform growth and cultured for two weeks were selected for the experiment (one seedling per pot). The synergistic microemulsion was diluted to 5 g / L with deionized water and sprayed onto the strawberry seedlings at a rate of 10 mL per seedling. Water spraying served as a control. Five strawberry seedlings were treated per group. Samples were taken from both the experimental and control groups on days 1, 3, and 7 after spraying. On day 8, 10 mL of the 5 g / L synergistic agent solution was sprayed again per seedling. Samples were taken again on days 9, 11, and 14. After grinding with liquid nitrogen, 0.1 g tissue samples were weighed and stored at 80℃ for later use. For the continuous spraying treatment group, 5 g / L synergistic agent solution was sprayed on days 1, 3, and 5. Samples were taken on days 1, 3, 7, 9, 11, and 14 after the fifth application. Water spraying served as a control. This was used to determine the activity of defensive enzymes and the expression levels of flavonoid pathway genes. Results are as follows: Figure 6 , 7 As shown in the figure. The results indicated that when the synergist was administered twice at 7-day intervals, the activity of the defense enzyme exhibited a clear bimodal fluctuation, showing a trend of first increasing and then decreasing over time. The peak value after the second stimulation was higher than that after the first stimulation. The expression of key genes in the flavonoid pathway was induced to show a clear bimodal high expression pattern, with the peak values ​​corresponding to the induction phase after the two synergist administrations. When the synergist was administered three times consecutively at 3-day intervals, the enzyme activity peaked 3 days after administration, and then the enzyme activity tended to stabilize. The expression levels of each gene remained at a stable level overall. An effective induction peak was formed in the early stage, and then it returned to a stable state.

[0060] 5. Interaction mechanism of components in synergists Tannic acid, quercetin, and rutin were dissolved in DMSO in a ratio of 12:0.5:0.5, diluted with deionized water, and Tween 80 was added to prepare a 5 g / L aqueous solution, denoted as T1.

[0061] Tannic acid, quercetin, and rutin were dissolved in DMSO in a ratio of 10:1:1, diluted with deionized water, and Tween 80 was added to prepare a 5 g / L aqueous solution, denoted as T2.

[0062] Tannic acid, quercetin, and rutin were dissolved in DMSO in a ratio of 5:1:1, diluted with deionized water, and Tween 80 was added to prepare a 5 g / L aqueous solution, denoted as T3.

[0063] Dissolve pure tannic acid in deionized water to prepare 5 g / L and 1 g / L tannic acid aqueous solutions. Quercetin was dissolved in DMSO to prepare a 5 g / L stock solution, which was then diluted with deionized water and Tween 80 was added to aid dissolution, resulting in a 1 g / L aqueous solution.

[0064] Dissolve rutin in DMSO to prepare a 5 g / L stock solution, dilute with deionized water, add Tween 80 to aid dissolution, and prepare a 1 g / L aqueous solution.

[0065] Fresh cowpea leaves were selected and cut into 2cm diameter leaf discs. Each disc was immersed in a solution corresponding to a specific treatment (tannin, quercetin, rutin, or combinations thereof in different proportions) for 10 seconds, then removed and air-dried. A layer of water agar was poured to the bottom of each petri dish. The group and number of replicates were marked on the lid and bottom of the dish with a marker. The treated leaves and corresponding control leaves were laid flat in the petri dish, overlapping each other to ensure their position was fixed. Healthy adult mites were selected from a pool of mites using a brush. 30 mites were introduced into each petri dish, and 15 mites were placed in each leaf disc. The petri dish lids were quickly closed. Each treatment group was repeated in triplicate. The start time of the experiment was recorded. At 24h and 48h after the start of the experiment, the number of mites on the two leaves in each petri dish was observed and recorded. The number of mites on the control and treated leaves at each time point was counted, and the repulsion rate was calculated. The results are shown below. Figure 8 As shown.

[0066] A contact-avoidance experiment was conducted using the leaf selection method. Strawberry leaves were collected after being sprayed with a 5 g / L tannic acid aqueous solution, with potted plants sprayed with plain water serving as a control. 8 g of agar strips were weighed, placed in a bottle, and 400 mL of deionized water was added. The bottle cap was gently screwed on, but not tightly, and placed in a microwave oven on medium-high for 30 minutes until the agar strips were completely melted and mixed thoroughly. 15 mL of water agar was poured into each petri dish, and after cooling and solidification, filter paper was moistened with deionized water and laid flat on the culture medium. Strawberry leaves were cut at days 1, 3, 5, and 8 after tannic acid induction. Following the principle of a control group on the left and a treatment group on the right, the strawberry leaves were laid flat on the agar with the back facing up, two leaves touching. The petioles and leaf margins were wrapped with moistened absorbent cotton to prevent wilting. Then, using a paintbrush, 20 starved two-spotted spider mites were inoculated onto each leaf. The leaves were covered with a petri dish and placed in an incubator at (25±1)℃, (45±5)% relative humidity, and a photoperiod of (16L:8D) for 2 hours. Inactive two-spotted spider mites were removed, and the leaves were replenished and placed under the same conditions. The number of two-spotted spider mites on each leaf was observed and recorded under a stereomicroscope after 24 and 48 hours. The avoidance rate was calculated.

[0067] Repellency rate = [(Number of leaf-dwelling insects in the control group - Number of leaf-dwelling insects in the treatment group) / Total number of insects] × 100% Depend on Figure 8It was found that after 24 h and 48 h of treatment, when tannic acid, quercetin, and rutin were applied individually to the two-spotted spider mite, tannic acid had the lowest repellency rate, while quercetin and rutin had significantly higher repellency rates. When the three substances were combined in a certain ratio, their repellency rates were significantly higher than those achieved by treating each substance individually. The highest repellency rate (75.57%) was achieved when the ratio of tannic acid:quercetin:rutin was 10:1:1, followed by a ratio of 12:0.5:0.5 (66.12%). This indicates that the individual effects of tannic acid, quercetin, and rutin are lower than their combined effects, and that a combination ratio within the range of 12:0.5:0.5 to 10:1:1 yields the best results.

[0068] like Figure 9 As shown, at 24 h, the repellency rates of the two-spotted spider mite in the leaves induced by tannic acid were 10.7%, 15.3%, 18.7%, and 13.7%, respectively, with the highest repellency rate on the 5th day; at 48 h, the repellency rates of the two-spotted spider mite in the leaves treated with tannic acid were 6.3%, 23.3%, 27.7%, and 15.7%, respectively, reaching the maximum on the 5th day; only on the 3rd and 5th days after induction, the repellency rate at 48 h was significantly higher than that at 24 h.

[0069] Quercetin and rutin primarily function as repellents, while tannic acid exerts a comprehensive effect, inducing resistance, deterring feeding, and repelling pests, with superior efficacy at higher proportions. Therefore, in summary, when synergists are applied, tannic acid can induce insect-resistant defense responses in plants, increase flavonoid content, and accumulate endogenous quercetin and rutin, enhancing the plant's endogenous repellent basis; it also promotes cell wall thickening, forming a dual physical and chemical defense. However, the feeding-repelling effect of tannic acid alone is easily degraded by insect detoxification enzymes. Quercetin and rutin, on the other hand, exert repellent effects, deterring adult insects from laying eggs and reducing damage from the next generation of larvae. Simultaneously, they can inhibit the detoxification activities of insect carboxylesterase and glutathione S-transferase, reducing insect tolerance to tannic acid and further enhancing its feeding-repelling effect. The combination of these three can mutually promote each other, further strengthening the inhibitory effect on pests.

[0070] Example 2

[0071] Tannic acid 12%, rutin 0.5%, quercetin 0.5%, dimethyl sulfoxide 10%, cyclohexanone 10%, alkylphenol polyoxyethylene ether-10 (OP-10) 4%, calcium dodecylbenzenesulfonate (agricultural emulsion 500#) 4.5%, polyvinyl alcohol 2%, sodium citrate 0.1%, ethylene glycol 1%, water to 100%.

[0072] Example 3

[0073] Tannic acid 5%, rutin 1%, quercetin 1%, dimethyl sulfoxide 10%, cyclohexanone 10%, alkylphenol polyoxyethylene ether-10 (OP-10) 4%, calcium dodecylbenzenesulfonate (agricultural emulsion 500#) 4.5%, polyvinyl alcohol 2%, sodium citrate 0.1%, ethylene glycol 1%, water to 100%.

[0074] The synergists prepared in Examples 1-3 were diluted with deionized water to prepare an aqueous solution of synergists at a concentration of 5 g / L.

[0075] Dissolve the pure tannic acid in deionized water to prepare a 1 g / L tannic acid aqueous solution. Quercetin was dissolved in DMSO to prepare a 5 g / L stock solution, which was then diluted with deionized water and Tween 80 was added to aid dissolution, resulting in a 1 g / L aqueous solution.

[0076] Dissolve rutin in DMSO to prepare a 5 g / L stock solution, dilute with deionized water, add Tween 80 to aid dissolution, and prepare a 1 g / L aqueous solution.

[0077] Commercial 30% etoxazole suspension was diluted with deionized water to prepare aqueous solutions of 75 mg / L and 100 mg / L.

[0078] Commercial 20% acetamiprid soluble solution was diluted with deionized water to prepare aqueous solutions of 18.75 mg / L and 25 mg / L.

[0079] Water was used as a blank control.

[0080] Application Implementation: (1) Potted strawberry seedlings with uniform growth and cultured for two weeks were selected for the experiment (one seedling per pot). A synergist diluted with deionized water (5 g / L, 10 mL per pot) was sprayed, as was a tannic acid / quercetin / rutin solution (1 g / L, 10 mL per pot). Ethoxyfenozide (75 mg / L and 100 mg / L, 10 mL per pot) was sprayed. A water control was set up. Each strawberry seedling was inoculated with 80 two-spotted spider mites. After 3 days, once the insect population stabilized, etoxazole was sprayed. Six days later, a synergist, tannic acid, quercetin, and rutin solution (10 mL per pot) was sprayed. Each treatment had three replicates, with three plants per replicate, and distance was maintained between replicates. The number of test insects on the leaves of each plant was investigated one day before and seven days after application. The number of surviving insects in each treatment was recorded, and the control effect was calculated based on the experimental data.

[0081] (2) Potted cucumber seedlings with uniform growth and cultured for two weeks were selected for the experiment (one seedling per pot). A treatment was applied by spraying 10 mL of synergist diluted with deionized water at a concentration of 5 g / L per pot, and a treatment was applied by spraying 10 mL of tannic acid / quercetin / rutin at a concentration of 1 g / L per pot. Acetamiprid at concentrations of 18.75 mg / L and 25 mg / L per pot was also applied at a concentration of 10 mL per pot. A water control was set up. Fifty aphids were introduced into each strawberry seedling. After 3 days, once the insect population stabilized, acetamiprid was sprayed. Six days later, a treatment was applied using 10 mL of a solution of synergist, tannic acid, quercetin, and rutin per pot. Each treatment had three replicates, with three plants per replicate, and the replicates were kept at a distance from each other. The number of insects on the leaves of each plant was investigated one day before and seven days after application. The number of surviving insects in each treatment was recorded, and the control effect was calculated based on the experimental data. The results are shown in Tables 10 and 11.

[0082] Table 10. Control efficacy of different synergists combined with etoxazole for 7 days against strawberry two-spotted spider mite.

[0083] Table 11. Control efficacy of different synergist components combined with acetamiprid against cucumber aphids after 7 days of treatment.

[0084] Tables 10 and 11 show that the control effects of tannic acid, quercetin, and rutin on strawberry two-spotted spider mites and cucumber aphids were significantly higher when combined with chemical agents than when the chemical agents were used alone. Among these, the control effect of tannic acid combined with chemical agents was superior to that of quercetin or rutin alone. Furthermore, the control effect of mixing the three secondary metabolites and combining them with chemical agents was superior to that of any single secondary metabolite combined with a chemical agent.

[0085] Example 4

[0086] The effect of synergists on chemical pesticides The synergist microemulsion formulation is as follows: 10% tannic acid, 1% rutin, 1% quercetin, 10% dimethyl sulfoxide, 10% cyclohexanone, 4% alkylphenol polyoxyethylene ether-10 (OP-10), 4.5% calcium dodecylbenzenesulfonate (agricultural emulsion 500#), 2% polyvinyl alcohol, 0.1% sodium citrate, 1% ethylene glycol, and water to make up to 100%.

[0087] Dilute the synergistic microemulsion to a concentration of 5 g / L with deionized water.

[0088] Commercial 22.4% spirotetramat suspension was diluted with deionized water to a 48 mg / L aqueous solution.

[0089] Water was used as a blank control.

[0090] Application Implementation: Potted strawberry seedlings with uniform growth, cultivated for two weeks, were selected for the experiment (one seedling per pot). Spirotetramat 48 mg / L was sprayed at 10 mL per pot. Three days later, a synergist diluted with deionized water (5 g / L) was sprayed at 10 mL per pot, and 10 mL was applied as a root drench treatment per pot. A water control was included. The soil was kept moist by regular watering. Leaf samples were collected at 1, 4, 7, and 10 days after application and stored at -80℃. The spirotetramat concentration was quantitatively determined using UPLC-MS / MS.

[0091] Depend on Figure 10 It was found that the content of spirotetramat under the three treatments showed a dynamic trend of first increasing and then decreasing, reaching its peak in all treatments 4 days after application, and then slowly degrading with plant metabolism. The control group without synergist had the lowest spirotetramat accumulation level. Both foliar spraying and root drenching with synergist increased the accumulation of spirotetramat, with foliar spraying showing a higher effect than root drenching. This indicates that foliar application of synergist can significantly prolong the residual effect of spirotetramat on leaves. In summary, synergist, whether applied foliarly or in combination with root drenching, can promote the accumulation of the pesticide in leaves and prolong its residual effect.

Claims

1. A pesticide synergist, characterized in that, Its active ingredients consist of tannic acid, rutin, and quercetin; the mass percentage of the active ingredients in the pesticide synergist is: tannic acid 10%-12%, rutin 0.5%-1%, and quercetin 0.5%-1%. The pesticide synergist can reduce the amount of chemical pesticides used by 25%.

2. The pesticide synergist according to claim 1, characterized in that, Its dosage form is a microemulsion.

3. The pesticide synergist according to claim 1, characterized in that, The pesticide synergist also includes 10% dimethyl sulfoxide, 10% cyclohexanone, and 4% alkylphenol polyoxyethylene ether-10. Calcium dodecylbenzenesulfonate 4.5%, polyvinyl alcohol 2%, sodium citrate 0.1%, ethylene glycol 1%, water to 100%.

4. A method for enhancing the efficacy of pesticides, characterized in that, include: The chemical agent and the pesticide synergist as described in any one of claims 1-3 are applied to the plant in sequence, with the pesticide synergist applied on the third day after the chemical agent is applied.

5. The pesticide enhancement method according to claim 4, characterized in that, The pesticide synergist is applied by foliar spraying or root irrigation.

6. The pesticide enhancement method according to claim 4, characterized in that, Applying the synergist a second time after a 7-day interval can enhance the control effect to a higher level 14 days after application.

7. The pesticide enhancement method according to claim 4, characterized in that, The application concentration of pesticide synergists is 2-5 g / L.

8. The pesticide enhancement method according to claim 4, characterized in that, The chemical agent is used to control one of the following: two-spotted spider mite, diamondback moth, aphid, or whitefly.

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