Application of Inhibiting Farnesol Dehydrogenase in Enhancing the Toxicity of Trifluoxetine

Abstract

This invention provides the application of inhibiting farnesol dehydrogenase in enhancing the toxicity of trifluralin, belonging to the field of pest control technology. This invention provides the application of inhibiting farnesol dehydrogenase in enhancing the toxicity of trifluralin. This invention improves the toxicity of trifluralin and enhances its control effect on pests by inhibiting the expression of farnesol dehydrogenase. Results from examples show that silencing farnesol dehydrogenase in Aedes aegypti using dsRNA is effective. FDL22 and peach aphid farnesol dehydrogenase FDL242 The expression of [specific ingredient] significantly enhances the toxicity of trifluralin against Aedes aegypti and the peach aphid. The combined use of the juvenile hormone analog S-(+)-tebufenozide with trifluralin significantly increases the toxicity of trifluralin against Aedes aegypti larvae.

Description

Technical Field

[0001] This invention relates to the field of pest control technology, specifically to the application of inhibiting farnesol dehydrogenase in enhancing the toxicity of trifluralin. Background Technology

[0002] Insecticides play an irreplaceable core role in the control of pests in the field. Current research and application of nematicides and acaricides not only aim to precisely control harmful nematodes, insects, and mites and address existing pesticide resistance issues in the field, but also to consider environmental safety to promote sustainable agricultural development. Therefore, the creation and development of insecticides with novel targets and modes of action are crucial. Trifluralin is a novel, green nematicide and acaricide independently developed in my country that combines nematicide and acaricide properties. It exhibits no cross-resistance with common nematicides and acaricides on the market, has long-lasting and stable efficacy, is green, low in toxicity, leaves no residue, and has high ecological safety, making its application prospects broad.

[0003] Research on the toxicity synergy of insecticides is a key area of ​​innovation in agricultural science and technology. The aim is to explore ways to reduce insecticide usage while effectively killing pests, thereby conserving resources, reducing pollution, delaying the development of insecticide resistance, and extending the lifespan of existing insecticides. Currently, several invention patents have been published regarding the toxicity synergy of trifluralin (CN113875758B, CN110946146B, CN119791116A, CN109006821A, etc.). Most of these studies focus on the combined application of trifluralin with other commercially available insecticides, which limits their scope of application. Therefore, in-depth research into the toxicity synergy pathways and mechanisms of trifluralin will contribute to its widespread promotion and application, and also provide fundamental support for research on pesticide compounding and synergistic effects. Summary of the Invention

[0004] This invention provides the application of inhibiting farnesol dehydrogenase in enhancing the toxicity of trifluralin. By inhibiting the expression of farnesol dehydrogenase in pests, the toxicity of trifluralin to pests can be significantly improved.

[0005] This invention provides the application of inhibiting farnesol dehydrogenase in enhancing the toxicity of trifluorometholone.

[0006] Preferably, the farnesol dehydrogenase comprises any one of the following (1) to (3):

[0007] (1) The amino acid sequence of Aedes aegypti farnesol dehydrogenase FDL22 is shown in SEQ ID NO.1; the nucleotide sequence encoding the protein of Aedes aegypti farnesol dehydrogenase FDL22 is shown in SEQ ID NO.2;

[0008] (2) The amino acid sequence of the peach aphid farnesol dehydrogenase FDL242 is shown in SEQ ID NO.13; the nucleotide sequence encoding the peach aphid farnesol dehydrogenase FDL242 protein is shown in SEQ ID NO.14;

[0009] (3) Homologous genes of (1) or (2) in other insects except Aedes aegypti and Prunus persica.

[0010] Preferably, methods for inhibiting farnesol dehydrogenase expression include knocking down farnesol dehydrogenase with dsRNA and / or inhibiting farnesol dehydrogenase expression by topical application of juvenile hormone analogs.

[0011] Preferably, the juvenile hormone analogue includes S-(+)-tebufenozide.

[0012] This invention provides a farnesol dehydrogenase from Aedes aegypti mosquitoes. FDL22 The dsRNA, the nucleotide sequence of which is shown in SEQ ID NO.3.

[0013] This invention provides a peach aphid farnesol dehydrogenase FDL242 The dsRNA, the nucleotide sequence of which is shown in SEQ ID NO.15.

[0014] This invention provides a synergistic insecticide comprising dsRNA that inhibits farnesol dehydrogenase expression and trifluralin.

[0015] Preferably, the dsRNA that inhibits farnesol dehydrogenase expression includes farnesol dehydrogenase from Aedes aegypti. FDL22 dsRNA or peach aphid farnesol dehydrogenase FDL242 dsRNA.

[0016] This invention provides the application of the dsRNA or the synergistic insecticide described in the above-mentioned technical solutions in the control of pests.

[0017] This invention provides a method for controlling pests using the synergistic insecticide described in the above-mentioned technical solution, comprising:

[0018] When the synergistic insecticide includes dsRNA that inhibits farnesol dehydrogenase expression, the dsRNA is delivered into the pest, and the pest is then exposed to trifluralin.

[0019] This invention provides the application of inhibiting farnesol dehydrogenase in enhancing the toxicity of trifluralin. This invention significantly improves the toxicity of trifluralin and enhances its control effect on pests by inhibiting the expression of farnesol dehydrogenase in insects. The results of the embodiments of this invention show that: knocking down the expression of farnesol dehydrogenase FDL22 in Aedes aegypti via dsRNA significantly increases the toxicity of trifluralin against Aedes aegypti larvae; by using the juvenile hormone analog S-(+)-tebufenozide in combination with trifluralin via external application, the expression of farnesol dehydrogenase FDL22 in Aedes aegypti can be inhibited, and the mixed solution has significantly improved the toxicity of trifluralin against Aedes aegypti larvae compared to trifluralin alone; by knocking down the homologous gene of farnesol dehydrogenase FDL22 in peach aphids, the expression of farnesol dehydrogenase in peach aphids can be enhanced. FDL242 This can significantly enhance the toxicity of trifluralin against peach aphid nymphs. Attached Figure Description

[0020] Figure 1 Feeding and soaking Aedes aegypti larvae FDL22 Significantly reduces farnesol dehydrogenase in Aedes aegypti larvae FDL22 Graph showing mRNA expression levels;

[0021] Figure 2 Farnesol dehydrogenase in Aedes aegypti mosquito larvae FDL22 The effect of inhibiting mRNA expression levels on the toxicity enhancement of 133.30 mg / L trifluralin against Aedes aegypti larvae is shown in the figure.

[0022] Figure 3 The toxicity enhancement effect of a mixture of 3 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin on Aedes aegypti larvae after 48 h of treatment is shown in the figure.

[0023] Figure 4 The toxicity enhancement effect of a mixture of 6 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin on Aedes aegypti larvae after 24 h of treatment is shown in the figure.

[0024] Figure 5 The toxicity enhancement effect of a mixture of 10 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin on Aedes aegypti larvae after 24 h of treatment is shown in the figure.

[0025] Figure 6 S-(+)-tebufenozide and trifluralin were used in combination to inhibit farnesol dehydrogenase. FDL22 Graph showing mRNA expression levels;

[0026] Figure 7A schematic diagram showing the amino acid sequence comparison between the farnesol dehydrogenase FDL22 of Aedes aegypti mosquito and the farnesol dehydrogenase FDL242 of Prunus sylvestris aphid.

[0027] Figure 8 To inject ds into peach aphid nymphs FDL242 Significantly reduced peach aphid nymphs FDL242 Graph showing mRNA expression levels;

[0028] Figure 9 farnesol dehydrogenase in peach aphid nymphs FDL242 The effect of inhibiting mRNA expression levels on the toxicity enhancement of 128.64 mg / L trifluralin solution against peach aphid nymphs is shown in the figure.

[0029] Figure 10 farnesol dehydrogenase in peach aphid nymphs FDL242 The effect of decreased mRNA expression levels on the toxicity of 230.16 mg / L trifluralin solution on peach aphid nymphs is illustrated in the figure.

[0030] Figure 11 A schematic diagram illustrating how inhibiting the expression of farnesol dehydrogenase enhances the toxicity of trifluralin to pests. Detailed Implementation

[0031] This invention provides the application of inhibiting farnesol dehydrogenase in enhancing the toxicity of trifluralin. As an optional embodiment of this invention, the farnesol dehydrogenase includes any one of the following (1) to (3): (1) Aedes aegypti farnesol dehydrogenase FDL22 with an amino acid sequence as shown in SEQ ID NO.1; and the nucleotide sequence encoding the protein of Aedes aegypti farnesol dehydrogenase FDL22 as shown in SEQ ID NO.2; (2) Peach aphid farnesol dehydrogenase FDL242 with an amino acid sequence as shown in SEQ ID NO.13; and the nucleotide sequence encoding the protein of Peach aphid farnesol dehydrogenase FDL242 as shown in SEQ ID NO.14; (3) the homologous gene of (1) or (2) in other pests other than Aedes aegypti and Peach aphid.

[0032] In this invention, the farnesol dehydrogenase comprises Aedes aegypti farnesol dehydrogenase FDL22, with the amino acid sequence shown in SEQ ID NO.1. As an optional embodiment of this invention, Aedes aegypti farnesol dehydrogenase... FDL22 The nucleotide sequence of the gene is shown in SEQ ID NO.2. In this invention, the Aedes aegypti farnesol dehydrogenase... FDL22 This refers to the gene encoding the farnesol dehydrogenase FDL22 protein in Aedes aegypti. This invention inhibits farnesol dehydrogenase in Aedes aegypti. FDL22 Expression can significantly enhance the toxicity of trifluralin against Aedes aegypti mosquitoes.

[0033] In this invention, the farnesol dehydrogenase comprises the peach aphid farnesol dehydrogenase FDL242, with the amino acid sequence shown in SEQ ID NO. 13. As an optional embodiment of this invention, the peach aphid farnesol dehydrogenase... FDL242 The nucleotide sequence of the gene is shown in SEQ ID NO.14. The gene encoding the peach aphid farnesol dehydrogenase FDL242 protein can also be called peach aphid farnesol dehydrogenase. FDL242 This invention inhibits farnesol dehydrogenase in peach aphids. FDL242 Expression can significantly enhance the toxicity of trifluralin against peach aphids.

[0034] In this invention, the farnesol dehydrogenase comprises a homologous gene of (1) or (2) found in other pests besides Aedes aegypti and Prunus persica. As an optional embodiment of this invention, the homologous gene preferably has an amino acid sequence similarity ≥40% with Aedes aegypti farnesol dehydrogenase FDL22 or Prunus persica farnesol dehydrogenase FDL242. This invention can enhance the toxicity of trifluralin to the corresponding insects by inhibiting the expression of the homologous gene in the respective insects.

[0035] As an optional embodiment of the present invention, the method for inhibiting farnesol dehydrogenase expression includes dsRNA knockdown of farnesol dehydrogenase and / or topical application of juvenile hormone analogs to inhibit farnesol dehydrogenase expression. In the present invention, the dsRNA includes farnesol dehydrogenase from Aedes aegypti. FDL22 dsRNA or peach aphid farnesol dehydrogenase FDL242 The dsRNA. As an optional embodiment of the present invention, the Aedes aegypti farnesol dehydrogenase... FDL22 The nucleotide sequence of the dsRNA is shown in SEQ ID NO.3; the peach aphid farnesol dehydrogenase FDL242 The nucleotide sequence of the dsRNA is shown in SEQ ID NO. 15. In this invention, the juvenile hormone analogue includes S-(+)-tebufenozide.

[0036] In this invention, when the juvenile hormone analog S-(+)-tebufenozide enhances the toxicity of trifluralin, the S-(+)-tebufenozide and trifluralin can be used together as a synergistic insecticide. Specifically, the synergistic insecticide includes S-(+)-tebufenozide and trifluralin. As an optional embodiment of the present invention, the weight ratio of the trifluralin to the S-(+)-tebufenozide can be ≥13:1, (13~65):1, (13~45):1, or 13:1, 15:1, 16:1, 18:1, 20:1, 22:1, 24:1, 26:1, 28:1, 30:1, 32:1, 34:1, 36:1, 38:1, 40:1, 41:1, 42:1, 43:1, 44:1, 44.4:1, 45:1, 46:1, 47:1, 48:1, 49... :1, 50:1, 51:1, 52:1, 53:1, 54:1, 55:1, 56:1, 57:1, 58:1, 59:1, 60:1, 61:1, 62:1, 63:1, 64:1, 65:1, 40:2, 45:2, 50:2, 55:2, 60:2, 65:2, 40:3, 45:3, 50:3, 55:3, 60:3, 65:3, 40:4, 45:4, 50:4, 55:4, 60:4, 65:4, 133.30:3, 133.30:6 or 133.30:10.

[0037] This invention relates to a method for controlling pests using a synergistic insecticide composed of S-(+)-tebufenozide and trifluralin, comprising: exposing pests to a solution containing a juvenile hormone analog and trifluralin. As an optional embodiment of this invention, when trifluralin and S-(+)-tebufenozide are combined, their weight ratio can be ≥13:1, (13~65):1, or (13~45):1. When using the synergistic insecticide obtained by combining trifluralin and S-(+)-tebufenozide to control pests, the exposure method can be immersing the pests in a solution containing the synergistic insecticide; the mass concentration of trifluralin in the solution containing the synergistic insecticide can be 133.30 mg / L; and the mass concentration of S-(+)-tebufenozide can be 3, 6, or 10 mg / L.

[0038] Aedes aegypti Aedes aegypti Larvae live in small bodies of water, their activity is restricted, and they are easily treated with pesticides, making them excellent models for toxicological research. In this invention, the Aedes aegypti mosquito larvae are used as a research model. Focusing on the conserved hormone in insects—juvenile hormone—and its synthetic pathway, the aim is to find ways and targets that can significantly enhance the efficacy of trifluralin. This is illustrated using the agricultural pest, the peach aphid. Myzus pericae These application examples provide support for the widespread use of trifluorometholone.

[0039] This invention provides a farnesol dehydrogenase from Aedes aegypti mosquitoes. FDL22 The dsRNA provided by this invention has the nucleotide sequence shown in SEQ ID NO.3. The dsRNA can efficiently knock down farnesol dehydrogenase in Aedes aegypti mosquitoes. FDL22 The expression of farnesol dehydrogenase FDL22 in Aedes aegypti mosquitoes is silenced. In this invention, the farnesol dehydrogenase... FDL22 dsRNA can also be called ds FDL22 .

[0040] This invention provides a peach aphid farnesol dehydrogenase FDL242 The dsRNA provided by this invention has the nucleotide sequence shown in SEQ ID NO. 15. This dsRNA can efficiently knock down farnesol dehydrogenase in peach aphids. FDL242 The expression of the peach aphid farnesol dehydrogenase FDL242 is silenced. In this invention, the peach aphid farnesol dehydrogenase... FDL242 dsRNA can also be called ds FDL242 .

[0041] This invention provides a synergistic insecticide comprising dsRNA that inhibits farnesol dehydrogenase expression and trifluralin.

[0042] As an optional embodiment of the present invention, the dsRNA includes farnesol dehydrogenase from Aedes aegypti. FDL22 dsRNA or peach aphid farnesol dehydrogenase FDL242 The dsRNA. As an optional embodiment of the present invention, the Aedes aegypti farnesol dehydrogenase... FDL22 The nucleotide sequence of the dsRNA is shown in SEQ ID NO.3; the peach aphid farnesol dehydrogenase FDL242 The nucleotide sequence of the dsRNA is shown in SEQ ID NO.15.

[0043] This invention provides the application of the dsRNA or the synergistic insecticide described in the above-mentioned technical solutions in the control of pests. The dsRNA significantly increases the toxicity of trifluralin, improving the control effect against pests. In this invention, the dsRNA or juvenile hormone analog in the synergistic insecticide that inhibits farnesol dehydrogenase expression can inhibit farnesol dehydrogenase, thereby synergistically enhancing the pest control ability with trifluralin and improving the pest control effect. As an optional embodiment of this invention, the pests include mosquitoes and / or aphids; the mosquitoes include Aedes aegypti; the Aedes aegypti larvae include first-instar and / or second-instar larvae; the aphids include peach aphids; the peach aphids include adult aphids and nymphs.

[0044] This invention provides a method for controlling pests using the synergistic insecticide described above, comprising:

[0045] When the synergistic insecticide includes dsRNA that knocks down farnesol dehydrogenase expression, the dsRNA is delivered into the pest, and the pest is then exposed to a solution containing trifluralin.

[0046] As an optional embodiment of the present invention, when the synergistic insecticide comprises dsRNA that knocks down farnesol dehydrogenase expression; wherein the dsRNA comprises farnesol dehydrogenase from Aedes aegypti. FDL22 dsRNA or peach aphid farnesol dehydrogenase FDL242 The dsRNA. As an optional embodiment of the present invention, the Aedes aegypti farnesol dehydrogenase... FDL22 The nucleotide sequence of the dsRNA is shown in SEQ ID NO.3; the peach aphid farnesol dehydrogenase FDL242 The nucleotide sequence of the dsRNA is shown in SEQ ID NO. 15.

[0047] In this invention, when the dsRNA expressing farnesol dehydrogenase is knocked down, it is the farnesol dehydrogenase from Aedes aegypti. FDL22 When dsRNA is generated, the farnesol dehydrogenase of Aedes aegypti mosquitoes is... FDL22 The dsRNA is delivered into Aedes aegypti mosquitoes, and then the mosquitoes are exposed to a solution containing trifluralin. As an optional embodiment of the invention, the Aedes aegypti mosquitoes include first-instar and / or second-instar larvae. As an optional embodiment of the invention, the delivery method includes feeding and soaking. As an optional embodiment of the invention, the feeding and soaking method includes feeding the Aedes aegypti mosquitoes with food containing the dsRNA, and then soaking them in 0.1% DEPC-treated water containing the dsRNA after 24 hours. As an optional embodiment of the invention, the mass concentration of the dsRNA in the food containing the dsRNA can be 10 µg / mL; the mass concentration of the dsRNA in the 0.1% DEPC-treated water containing the dsRNA can be 50 µg / mL. After delivering the dsRNA into Aedes aegypti mosquitoes using the above feeding and soaking methods, the present invention detects... FDL22 The gene expression was significantly reduced, demonstrating that the dsRNA provided by this invention can knock down farnesol dehydrogenase in Aedes aegypti. FDL22 Silent Aedes aegypti farnesol dehydrogenase FDL22 Gene expression.

[0048] Knockdown of farnesol dehydrogenase in Aedes aegypti FDL22Following gene knockdown, the present invention exposes the *Aedes aegypti* mosquitoes to a solution containing trifluoperamide. As an optional embodiment of the present invention, the exposure includes immersion of the *Aedes aegypti* mosquitoes in a trifluoperamide solution for 48 hours. As an optional embodiment of the present invention, the mass concentration of the trifluoperamide solution is ≥133.30 mg / L. The present invention demonstrates through efficacy verification that it knocks down farnesol dehydrogenase in *Aedes aegypti* mosquitoes. FDL22 After gene expression, the toxicity of trifluralin to Aedes aegypti mosquito larvae was significantly enhanced.

[0049] In this invention, when the dsRNA inhibiting farnesol dehydrogenase expression is peach aphid farnesol dehydrogenase... FDL242 When dsRNA is generated, peach aphid farnesol dehydrogenase is used. FDL242 The dsRNA was delivered into the peach aphid, and then the aphid was exposed to the trifluralin. As an optional embodiment of the invention, the peach aphid includes adult and / or nymphal aphids. As an optional embodiment of the invention, the delivery method includes injection. The invention does not specifically limit the injection method; any conventional injection method in the art can be used. As an optional embodiment of the invention, the injection method can be microinjection; the injection site can be the midcoxal fossa of a fourth instar nymph of the peach aphid; the concentration of the dsRNA during injection is 10000 ng / μL; the injection volume of the dsRNA can be 30 nL / aphid; and the injection rate can be 30 nL / s. After the dsRNA was delivered into the peach aphid by injection, the peach aphid farnesol dehydrogenase was detected. FDL242 The expression of [a specific enzyme] was significantly reduced, demonstrating that the dsRNA provided by this invention can knock down farnesol dehydrogenase in peach aphids. FDL242 Silent peach aphid farnesol dehydrogenase FDL242 The expression.

[0050] The present invention preferably uses peach aphid farnesol dehydrogenase. FDL242 Two days after dsRNA delivery to peach aphids, the aphids were exposed to a solution containing trifluoperamide. As an optional embodiment of the invention, the mass concentration of trifluoperamide in the solution is ≥128.64 mg / L, and can be between 128.64 and 230.16 mg / L. As an optional embodiment of the invention, the exposure time can be 10 s.

[0051] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the accompanying drawings and embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0052] Example 1: This case study uses Aedes aegypti larvae as a model and employs dsRNA interference to knock down farnesol dehydrogenase in Aedes aegypti. FDL22 This enhances the toxicity of trifluralin to Aedes aegypti larvae.

[0053] The amino acid sequence of farnesol dehydrogenase FDL22 from Aedes aegypti is shown in SEQ ID NO.1, specifically: MDRWAGKVAVVTGASSGIGAAITTDLAKAGMVVVGLARRVERVEALKANLPESAKPRLHAVKCDVSKEEDITQVFKWVEEKFGGVDVLVNNAGILRQTDLLGTDNGQMLREVLDTNVMGLVLCSQKAYQSMKKRSVDGHIVHINSVVGHKVFDFPQSNIYPASKHAVTAITETMRNELRNAGSRIKVTSISPGVVRTEILPESIIEGGHSLLESEDISEAVLYVLGTPPRVQVHELTIKPVGEKF.

[0054] The gene encoding the farnesol dehydrogenase FDL22 protein in Aedes aegypti mosquitoes, i.e., farnesol dehydrogenase. FDL22The nucleotide sequence is shown in SEQ ID NO.2, specifically: 5’-atggaccgttgggcaggaaaggtggccgtagtgaccggggcaagttcgggaattggagccgccatcactacggacctggccaaagctggaatggtggtcgttggattggcacgtcgagtggaacgggttgaagctttgaaagcaaacttgccggagtctgctaagccgcgtttgcatgcggttaagtgtgacgtatccaaggaggaagacattacccaggtgttcaaatgggttgaggagaaatttgggggagttgacgtgttggtcaacaatgcgggaatccttcggcagacggatttgctgggtaccgataatggtcaaatgctgcgagaagtcttggatacgaatgtgatgggactggtgctctgcagccagaaggcctatcaatcgatgaagaagcgctcggtcgatgggcacattgtccatataaatagcgtcgttggccataaggtttttgatttcccgcaatcgaatatatatccggcgtcgaagcacgcagtgacggccattacggaaactatgagaaacgaattgcggaatgccgggtccagaataaaggttacgagtattagccccggtgtggttcgcactgaaatccttccggagtccattattgaaggaggtcattcgctgttggaatcggaagatatatccgaagctgtattgtacgtgcttggaacacctcctcgagtgcaagtccatgaattgactataaaaccagtaggtgagaagttttga-3’。

[0055] 1. FDL22 Synthesis of dsRNA Query the mRNA sequence (named FDL22siDirect (https: / / sidirect2.rnai.jp / ) was used for siRNA prediction. Regions with as many siRNAs as possible and low off-target efficiency were selected as dsRNA templates. The template sequence is SEQ ID NO.3, specifically: 5'-tgggttgaggagaaatttgggggagttgacgtgttggtcaacaatgcgggaatccttcggcagacggatttgctgggtaccgataatggtcaaatgctgcgagaagtcttggatacgaatgtgatgggactggtgctctgcagccagaaggcctatcaatcgatgaagaagcgctcggtcgatgggcacattgtccatataaatagcgtcgtcgttggccataaggtttttgatttcccgcaatcgaatatata tccggcgtcgaagcacgcagtgacggccattacggaaactatgagaaacgaattgcggaatgccgggtccagaataaaggttacgagtattagccccggtgtggttcgcactgaaatccttccggagtcca ttattgaaggaggtcattcgctgttggaatcggaagatatatccgaagctgtattgtacgtgcttggaacacctcctcgagtgcaagtccatgaattgactataaaaccagtaggtgagaagttttga-3'.

[0056] Based on the dsRNA template sequence, primers with the T7 promoter were designed using the online primer design website (https: / / crm.vazyme.com / cetool / simple.html?enzymeLociFlag=Y). The primer sequences are as follows: ds FDL22 F: 5'-taatacgactcactatagggTGGGTTGAGGAGAAATTTGGG-3' (SEQ ID NO.4), ds FDL 22 R: 5'-taatacgactcactatagggTCAAAACTTCTCACCTACTGGTTTTATA-3' (SEQ ID NO. 5).

[0057] RNA was extracted from Aedes aegypti larvae tissue using the TransZol Up kit. After 1% agarose gel electrophoresis, concentration determination, and quality assessment, the extracted RNA passed quality checks. cDNA was then synthesized using the HiScript III 1st Strand cDNA Synthesis Kit (+gDNA wiper) (Novizan) with 1 µg of RNA as a template. The cDNA was then used as a template to amplify ds... FDL22 The template region was examined, and the PCR product was verified by 1% agarose gel electrophoresis. A single band was observed. DNA template was extracted using the phenol / chloroform method and purified by ethanol / sodium acetate precipitation. Using 1 µg of DNA as template, dsRNA was synthesized using the TranscriptAid T7 high-yield transcription kit (Thermo Fisher Scientific). Finally, dsRNA was extracted using the phenol / chloroform method and purified by ethanol / sodium acetate precipitation. After concentration determination by 1% agarose gel electrophoresis, the dsRNA was aliquoted and stored at -80°C.

[0058] 2. GFP dsRNA (named ds) GFP Synthesis of ) using primer ds GFP -F: 5'-taatacgactcactatagggAAGGGCGAGGAGCTGTTCACCG-3' (SEQ ID NO.6) and primer ds GFP -R: 5'-taatacgactcactatagggCAGCAGGACCATGTGATCGCGC-3' (SEQ ID NO.7) amplifies ds from a plasmid containing the GFP fragment. GFP The template region (SEQ ID NO. 8) was verified by 1% agarose gel electrophoresis of the PCR product. The band was single, and the DNA template was extracted using the phenol / chloroform method and purified by ethanol / sodium acetate precipitation. dsRNA was synthesized using the TranscriptAid T7 High-Yield Transcription Kit (Thermo Fisher Scientific) with 1 µg of DNA as the template. Finally, the dsRNA was extracted using the phenol / chloroform method and purified by ethanol / sodium acetate precipitation. After concentration determination by 1% agarose gel electrophoresis, the dsRNA was aliquoted and stored at -80°C.

[0059] The dsRNA of the GFP gene is called ds GFP As a comparison; FDL22 The dsRNA of a gene is called ds FDL22 .

[0060] The nucleotide sequence of SEQ ID NO.8 is: 5’-aagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccttcagctacggcgtgcagtgcttcagccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggccaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagccaccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctg-3’。

[0061] 3. Delivery of dsRNA ds GFP and ds s FDL22 were introduced into Aedes aegypti larvae using the following methods respectively.

[0062] dsRNA was delivered via a feeding and soaking method. An equal volume of 1% agarose was mixed with overnight fermented larval food (fish food: yeast = 6.4 mg / mL: 3.2 mg / mL) and placed in a 55°C water bath. 20 µg of dsRNA was added to 2 mL of the mixture, and then this mixture was used to fill one well of a six-well cell culture plate. After solidification, 40 newly hatched Aedes aegypti larvae were counted and introduced into one well. The next day, 20 µg of dsRNA was dissolved in 400 µL of 0.1% DEPC water and added to a single well. On the third day, half of the larvae from each well under different treatments were sampled to extract RNA to examine RNA interference efficiency. The other half of the larvae from each well were used for virulence experiments.

[0063] 4. Interference efficiency was detected using the TransZol Up kit (Full Gold) to analyze the interference efficiency of the ds... GFP and ds FDL22 RNA was extracted from the treated larvae. After 1% agarose gel electrophoresis, concentration determination, and quality assessment, the extracted RNA passed the quality test. Using the HiScript III 1st Strand cDNA Synthesis Kit (+gDNA wiper) (Novizan), cDNA was synthesized by reverse transcription using 1µg of RNA as a template. The synthesized cDNA was diluted 10-fold for quantitative real-time PCR. FDL22 Changes in expression levels. Primers used in quantitative real-time PCR were designed online (https: / / www.ncbi.nlm.nih.gov / tools / primer-blast / index.cgi?LINK_LOC=BlastHome), with the following sequence information: q-FDL22 F: 5'-TCGAGTGGAACGGGTTGAAG-3' (SEQ ID NO.9), q-FDL22 R: 5'-ACCTGGGTAATGTCTTCCTCCT-3' (SEQ ID NO.10); q-RSP17 F: 5'-AAGAAGTGGCCATCATTCCA-3' (SEQ ID NO.11), q-RSP17 R: 5'-GGTCTCCGGGTCGACTTC-3' (SEQ ID NO.12). qPCR reactions were performed on the Bio-Rad Real-Time PCR system using the ChamQ Universal SYBR qPCR MasterMix kit, with a 2... -ΔΔCt The qPCR data were processed using the method described in Table 1. The results are shown in Table 1 and... Figure 1 As shown. Figure 1 Feeding and soaking Aedes aegypti larvae FDL22Significantly reduces farnesol dehydrogenase in Aedes aegypti larvae FDL22 The graph shows the results of mRNA expression levels.

[0064] Table 1 ds GFP and ds FDL22 48 hours after treatment with Aedes aegypti mosquito larvae FDL22 relative expression level

[0065]

[0066] Note: n=4~6, indicating 4~6 biological replicates were performed; # indicates successful replication. t The test analysis showed a significant difference in the mean values ​​between the trifluralin + S-(+)-tebufenozide group and the trifluralin group. p <0.05), the same applies below.

[0067] From Table 1 and Figure 1 It can be seen that feeding and soaking delivery methods are used for ds FDL22 Aedes aegypti larvae FDL22 The expression level of dsGFP was reduced by approximately 40% compared to the control group of Aedes aegypti larvae delivered by feeding and soaking. This indicates that dsGFP expression can significantly interfere with Aedes aegypti mosquito larvae. FDL22 The expression of [the gene] achieved knockdown of FDL22 in Aedes aegypti larvae, satisfying the requirements for functional analysis and subsequent interference of Aedes aegypti larvae with dsRNA. FDL22 The requirements for the virulence enhancement experiment of trifluralin against Aedes aegypti larvae after expression.

[0068] 5. Toxicity test of Aedes aegypti larvae: The toxicity of second-instar larvae of Aedes aegypti that had not been exposed to trifluralin was determined. 25 larvae were subjected to ds... GFP The treated larvae and 25 ds FDL22 The treated larvae were immersed in 50 mL of trifluralin at a concentration of 133.30 mg / L for 48 h, and the change in mortality rate was used as the analytical indicator.

[0069] The results are shown in Table 2 and Figure 2 As shown. Figure 2 Farnesol dehydrogenase in Aedes aegypti mosquito larvae FDL22 The graph shows the synergistic effect of mRNA expression inhibition on the toxicity of 133.30 mg / L trifluralin against Aedes aegypti larvae.

[0070] Table 2 (by ds) GFP and ds FDL22 Mortality rate of Aedes aegypti larvae after 48 hours of treatment followed by immersion in a solution containing 133.30 mg / L trifluralin for 48 hours.

[0071]

[0072] From Table 2 and Figure 2 We can obtain, dsFDL22 The mortality rate of *Aedes aegypti* larvae pretreated for 48 h was approximately 20% higher after immersion in 133.30 mg / L trifluralin for 48 h compared to those pretreated with dsGFP for 48 h. This indicates that within knockdown *Aedes aegypti* larvae... FDL22 Under the condition of adequate expression, trifluralin can significantly enhance the toxicity of trifluralin to Aedes aegypti larvae. The expression level of FDL22 in Aedes aegypti is negatively correlated with the toxicity of trifluralin to larvae.

[0073] Example 2 This implementation case uses Aedes aegypti larvae as a model. By mixing and applying S-(+)-tebufenozide and trifluralin, the toxicity of trifluralin against Aedes aegypti larvae is significantly enhanced.

[0074] Using acetone as a solvent, a 3000 mg / L stock solution of S-(+)-tebufenozide was prepared. This stock solution was then added to 50 mL of a 133.30 mg / L trifluralin solution, resulting in final S-(+)-tebufenozide concentrations of 3 mg / L, 6 mg / L, and 10 mg / L in the mixed solutions. Toxicity was determined in second-instar larvae of *Aedes aegypti* that had not been exposed to trifluralin or S-(+)-tebufenozide. For each experimental group, 25 second-instar larvae were immersed in 50 mL of solutions containing 3 mg / L, 6 mg / L, and 10 mg / L S-(+)-tebufenozide mixed with 133.30 mg / L trifluralin, respectively. Larval mortality was recorded after 24 h or 48 h. Simultaneously, 3 mg / L, 6 mg / L, and 10 mg / L S-(+)-tebufenozide were added separately as control group 1, and 133.30 mg / L trifluralin was added separately as control group 2. The larval mortality rate of the experimental and control groups was used as the analysis index. The results are shown in Tables 3-5 and Figures 3-5 As shown, Figure 3 The toxicity enhancement effect of a mixture of 3 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin on Aedes aegypti larvae after 48 h of treatment is shown in the figure. Figure 4 The toxicity enhancement effect of a mixture of 6 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin on Aedes aegypti larvae after 24 h of treatment is shown in the figure. Figure 5 The toxicity enhancement effect of a mixture of 10 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin on Aedes aegypti larvae after 24 h of treatment is shown in the figure.

[0075] Table 3. Mortality rate (%) of larvae treated with 3 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin after 48 h.

[0076]

[0077] Note: n=5, 5 biological replicates were performed; different letters (a, b, c) indicate significant differences between groups as determined by one-way ANOVA and Tukey's test. p <0.05), the same applies below.

[0078] Table 4. Mortality rate (%) of larvae treated with 6 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin after 24 h.

[0079]

[0080] Table 5. Mortality rate (%) of larvae after 24 h of treatment with 10 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin.

[0081]

[0082] From Tables 3-5 and Figures 3-5 It was found that the combined use of 3 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin to treat Aedes aegypti larvae for 48 h increased toxicity by more than three times compared to trifluralin alone, while 3 mg / L S-(+)-tebufenozide had no toxic effect on Aedes aegypti larvae. Treatment of Aedes aegypti larvae with 6 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin for 24 h significantly increased toxicity compared to trifluralin alone. Treatment of Aedes aegypti larvae with 10 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin for 24 h also significantly increased toxicity compared to trifluralin alone. In summary, S-(+)-tebufenozide can significantly enhance the toxicity of trifluralin. When the two act on Aedes aegypti larvae, they produce a synergistic effect in the control of Aedes aegypti larvae.

[0083] Example 3: This case study uses Aedes aegypti larvae as a model to test the effects of S-(+)-tebufenozide and trifluralin on the growth of Aedes aegypti mosquitoes. FDL22 Downregulation exists.

[0084] A mixture of 6 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin was applied to 50 second-instar Aedes aegypti mosquito larvae for 2 days. RNA was extracted from surviving larvae using the TransZol Up kit. After 1% agarose gel electrophoresis, concentration determination, and quality assessment, the extracted RNA was used to reverse transcribe cDNA using the HiScript III 1st Strand cDNA Synthesis kit (+gDNA wiper) with 300 ng of RNA as a template. The synthesized cDNA was diluted 10-fold for quantitative real-time PCR. FDL22 Changes in expression levels. The primers and methods used for quantitative real-time PCR were the same as those used in the quantitative real-time PCR assay for interference efficiency in Example 1. The results of quantitative real-time PCR are shown below. Figure 6 See Table 6. Figure 6 S-(+)-tebufenozide and trifluralin were used in combination to inhibit farnesol dehydrogenase. FDL22 The graph shows the results of mRNA expression levels.

[0085] Table 6. Inhibition of S-(+)-acetamiprid and trifluralin mixture application FDL22 mRNA expression level

[0086]

[0087] From Table 6 and Figure 6 It was found that when 6 mg / L S-(+)-tebufenozide and 133.30 mg / L trifluralin were applied together, the concentration of tebufenozide in Aedes aegypti larvae was significantly reduced. FDL22 mRNA expression was downregulated; based on Case 2, dsRNA was used to interfere with Aedes aegypti mosquito larvae. FDL22 This enhances the toxicity of trifluralin against Aedes aegypti larvae. It has been demonstrated that Aedes aegypti larvae... FDL22 The mRNA level of trifluralin was negatively correlated with its toxicity. Therefore, the enhanced toxicity of trifluralin against Aedes aegypti larvae in the combined application of S-(+)-tebufenozide and trifluralin in Case Study 3 may also be due to the inhibition of Aedes aegypti mosquitoes. FDL22 The expression.

[0088] Example 4: Knockdown of the agricultural pest peach aphid using dsRNA interference FDL22 Homologous genes FDL242 It enhances the toxicity of trifluralin against peach aphids.

[0089] The peach aphid was identified by comparing the amino acid sequence of farnesol dehydrogenase FDL22 from the Aedes aegypti mosquito database with the Genbank database. M. persicaeThe gene encoding the homologous amino acid sequence is numbered LOC111037242 and named as follows: FDL242 This refers to the peach aphid farnesol dehydrogenase FDL242, which has a 44.90% amino acid sequence similarity to the Aedes aegypti farnesol dehydrogenase FDL22. Figure 7 This is a schematic diagram showing the amino acid sequence comparison between the farnesol dehydrogenase FDL22 of Aedes aegypti and the farnesol dehydrogenase FDL242 of Prunus persica.

[0090] The amino acid sequence of the peach aphid farnesol dehydrogenase FDL242 is shown in SEQ ID NO.13, specifically: MIFQIFSYCLDTGKHWSCSFYNIKIYIFKTKMEKWNGKVAVVTGASSGIGEETCRQLVERGMIVVGFARREDKLQVLENDLKGKLGKFYYVKVDLCSEENIMEAFNWVKSTLKSLDVLINNAGVLKQTDLLGSTKDWKQMFDTNVIGLNICSREAIKIMEQVQIKEGHIININSVAGHYQFPFMKDISVYSTTKHSVTIITESLRELLGMKNLPIRVTSISPAGVETEMTVELSKLEGFKMLKSIDIAEAILYALSAPQRVNVAEIIIRPTSENTLDFIKSLV.

[0091] The gene encoding farnesol dehydrogenase FDL242, also known as peach aphid farnesol dehydrogenase. FDL242The nucleotide sequence is shown in SEQ ID NO.14, specifically: 5’-atgatatttcagattttcagttattgtttggacaccggtaagcactggtcgtgtagtttttataatattaaaatatatatttttaaaacgaaaatggaaaagtggaatggaaaagttgctgtagtgacgggtgcttcttctggtatcggagaggagacttgtcgacaactggttgaaaggggaatgattgttgttggttttgctagaagagaagataaacttcaggtgttggaaaacgatttaaaaggaaaactgggtaaattttattacgttaaagttgatttatgttcggaagaaaacattatggaagcttttaattgggtaaagagtacattgaaatcgttagacgtactaattaacaatgctggtgttttgaagcaaactgatttattaggaagcacaaaagattggaaacaaatgtttgacactaatgtcataggacttaatatctgcagcagagaggccataaaaattatggagcaagtacaaatcaaagagggtcacataataaatattaacagtgttgcaggtcattatcaatttccatttatgaaggatatttcagtttatagcactaccaaacatagtgttactataatcacagagagtttaagagaattgctgggcatgaaaaatttacccatcagagtaacgagtatcagtccggccggagtagaaaccgaaatgactgtagagttaagcaaattggaaggatttaaaatgttgaaatctattgacattgctgaagctattttatacgcgttgagtgcaccgcaacgtgttaacgtcgccgaaatcatcattagaccaacaagtgaaaatacattagattttattaagagtcttgtgtag-3’。

[0092] 1. The LC of ethoprophos against the fourth instar nymphs of Myzus persicae 50The assay was performed according to the "People's Republic of China Agricultural Industry Standard NY / T 1154.6-2006 Pesticide Indoor Bioassay Test Guidelines Insecticides Part 6: Insecticide Activity Test - Immersion Method", determining the LC50 of trifluralin against fourth instar nymphs of the peach aphid. 50 Value. Each experimental group used 30 fourth-instar nymphs of the peach aphid. The results are shown in Table 7.

[0093] Table 7. Mortality rate (%) of peach aphid nymphs after treatment with different concentrations of trifluralin at different time points.

[0094]

[0095] Note: 1-3 represent 3 biological replicates.

[0096] 2. FDL242 The dsRNA synthesis was performed by querying the mRNA sequence based on the gene accession number LOC111037242 in the GenBank database. siDirect (https: / / sidirect2.rnai.jp / ) was used for siRNA prediction, selecting regions with the highest possible siRNA count and low off-target efficiency as dsRNA templates. The template sequence is SEQ ID NO.15, specifically: 5'-gacaactggttgaaaggggaatgattgttgttggttttgctagaagagaagataaacttcaggtgttggaaaacgatttaaaaggaaaactgggtaaattttattacgttaaagttgatttatgttcggaagaaaacattatggaagcttttaattgggtaaagagtacattgaaatcgttagacgtactaattaacaatgctggtgttttgaagcaaactgatttattaggaagcaaaagattgg aaacaaatgtttgacactaatgtcataggacttaatatctgcagcagagaggccataaaaattatggagcaagtacaaatcaaagagggtcacataataaatattaacagtgttgcaggtcattatcaatt tccatttatgaaggatatttcagtttatagcactaccaaacatagtgttactataatcacagagagtttaagagaattgctgggcatgaaaaatttacccatcagagtaacgagtatcagtccggcc-3'.

[0097] Based on the dsRNA template sequence, primers with the T7 promoter were designed using the online primer design website (https: / / crm.vazyme.com / cetool / simple.html?enzymeLociFlag=Y). The primer sequences are as follows: ds FDL242 F: 5'-taatacgactcactatagggGACAACTGGTTGAAAGGGGA-3' (SEQ ID NO. 16), ds FDL242 R: 5'-taatacgactcactatagggGGCCGGACTGATACTCGTTA-3' (SEQ ID NO. 17).

[0098] RNA was extracted from fourth-instar nymphs of the peach aphid using the TransZol Up kit. After 1% agarose gel electrophoresis, concentration determination, and quality assessment, the extracted RNA passed quality control. cDNA was synthesized using the HiScript III 1st Strand cDNA Synthesis Kit (+gDNA wiper) with 1 µg of RNA as a template. The cDNA was then used as a template to amplify ds... FDL242 The template region was examined, and the PCR product was verified by 1% agarose gel electrophoresis. A single band was observed. DNA template was extracted using the phenol / chloroform method and purified by ethanol / sodium acetate precipitation. Using 1 µg of DNA as template, dsRNA was synthesized using the TranscriptAid T7 high-yield transcription kit (Thermo Fisher Scientific). Finally, dsRNA was extracted using the phenol / chloroform method and purified by ethanol / sodium acetate precipitation. After concentration determination by 1% agarose gel electrophoresis, the dsRNA was aliquoted and stored at -80°C.

[0099] 3. GFP dsRNA (named ds) GFP The synthesis method of GFP is the same as in Example 1. The dsRNA of the GFP gene is referred to as dsRNA. GFP As a control, the sequence is referenced to SEQ ID NO.8 in Example 1; MpFDL242 The dsRNA of a gene is called ds FDL242 .

[0100] 4. Delivery of dsRNA: dsRNA is delivered separately. GFP and ds FDL242 The following methods were used to introduce dsRNA into the nymphs of the peach aphid.

[0101] dsRNA was delivered via microinjection. The previously synthesized dsRNA was then delivered... GFPand ds FDL242 Dilute to 10000 ng / μL with 0.1% DEPC water; select uniformly growing fourth-instar nymphs of the peach aphid and place them in 5 mL centrifuge tubes for later use; briefly stun the peach aphid nymphs with carbon dioxide and place them on a solidified 1% agarose gel plate, then use the Nanoinject III microinjection system to inject ds GFP or ds FDL242 The injection was administered into the nymphs of the peach aphid at the coxal fossa of the midlegs, with an injection volume of 30 nL per aphid and an injection rate of 30 nL / s. After injection, the peach aphids were placed in a petri dish containing fresh radish seedlings for rearing, and samples were taken at 24 h and 48 h to extract RNA to examine the RNA interference efficiency.

[0102] 5. Interference efficiency was detected using the TransZol Up kit (Full Gold) to analyze the interference efficiency of the ds-120 ... GFP and ds FDL242 RNA was extracted from treated peach aphid nymphs. The extracted RNA underwent 1% agarose gel electrophoresis, concentration determination, and quality assessment. After passing these tests, cDNA was synthesized using the HiScript III 1st Strand cDNA Synthesis Kit (+gDNA wiper) (Novizan) with 1 µg of RNA as a template. The synthesized cDNA was diluted 10-fold for quantitative real-time PCR detection. FDL242 Changes in expression levels. Primers used in quantitative real-time PCR were designed online (as in Example 1, section 4), and their sequence information is as follows: q-FDL242 F: 5'-GGTGCTTCTTCTGGTATCG-3' (SEQ ID NO.18), q-FDL242 R: 5'-CCCTCTTTGATTTGTACTTGC-3' (SEQ ID NO.19); quantitative internal control primers q-GAPDH F: 5'-ACGGACCATCTGGAAAATTATGGA-3' (SEQ ID NO.20), q-GAPDH R: 5'-GCAACTGGTACTCTGAAAGCC-3' (SEQ ID NO.21). qPCR reactions were performed on the Bio-Rad Real-Time PCR system using the ChamQ Universal SYBR qPCRMaster Mix kit, employing a 2... -ΔΔCt The qPCR data were processed using the method described above. The results are shown in Table 8 and... Figure 8 As shown. Figure 8 To inject ds into peach aphid nymphs FDL242 Significantly reduced peach aphid nymphs FDL242 The graph shows the results of mRNA expression levels.

[0103] Table 8 ds GFP and ds FDL242 24 and 48 hours after treatment of peach aphid nymphs FDL242 relative expression level

[0104]

[0105] From Table 8 and Figure 8 It can be seen that microinjection delivery of ds FDL242 peach aphid nymphs FDL242 The expression level compared to microinjection delivery of ds GFP The control group showed a reduction of approximately 64% at 24 hours and approximately 72% at 48 hours. This indicates that ds FDL242 It can significantly interfere with peach aphids FDL242 The expression was realized within the peach aphid nymph. FDL242 Knockdown was performed to meet the requirements of functional analysis and subsequent interference with peach aphid nymphs via dsRNA. FDL242 The requirements for the toxicity test of trifluralin against peach aphid nymphs after expression.

[0106] 6. Toxicity Experiment of Peach Aphid Nymphs: Toxicity of fourth-instar peach aphid nymphs that had not been exposed to trifluralin was determined using the leaf-dip method. 23 nymphs were injected with ds... GFP Larvae 48 hours later and 23 larvae injected with ds FDL242 After 48 hours, the nymphs were immersed in trifluralin at concentrations of 128.64 mg / L or 230.16 mg / L for 10 seconds, and then transferred to fresh radish seedlings for rearing. The mortality rate was recorded at 24 hours and 48 hours, and the change in mortality rate was used as the analytical indicator.

[0107] The results are shown in Tables 9-10 and... Figures 9-10 As shown. Figure 9 farnesol dehydrogenase in peach aphid nymphs FDL242 The effect of decreased mRNA expression levels on the toxicity enhancement of 128.64 mg / L trifluralin solution against peach aphid nymphs is shown in the figure. Figure 10 farnesol dehydrogenase in peach aphid nymphs FDL242 The graph shows the synergistic effect of decreased mRNA expression levels on the toxicity of 230.16 mg / L trifluralin solution against peach aphid nymphs.

[0108] Table 9. Mortality rate of peach aphid nymphs after treatment with 128.64 mg / L trifluralin.

[0109]

[0110] Table 10 Mortality rate of peach aphid nymphs after treatment with 230.16 mg / L trifluralin

[0111]

[0112] From Tables 9-10 and Figures 9-10 Therefore, ds FDL242 The mortality rate of peach aphid nymphs treated for 48 h was significantly lower than that of ds after 24 h and 48 h of treatment with 128.64 mg / L trifluralin compared to ds. GFP Treatment for 48 hours increased the number of peach aphid nymphs by approximately 2.90% and 11.60%. FDL242 The mortality rate of peach aphid nymphs treated with 230.16 mg / L trifluralin for 24 h and 48 h was lower than that of ds GFP Treatment for 48 hours increased the number of peach aphid nymphs by approximately 8.69% and 13.04%. This indicates that treatment effectively inhibits the growth of peach aphid nymphs. FDL242 Under the premise of expression, the toxicity of trifluralin to peach aphid nymphs can be significantly improved, and the expression level of FDL242 in peach aphids is negatively correlated with the toxicity of trifluralin to larvae.

[0113] In summary, the schematic diagram illustrating the enhancement of trifluoromethylpyrazine toxicity by inhibiting farnesol dehydrogenase expression in this invention is shown below. Figure 11 As shown. Aedes aegypti larvae ingest ds FDL22 Knockdown of FDL22 expression significantly enhanced the toxicity of trifluralin to Aedes aegypti larvae; peach aphid nymphs ingesting ds FDL242 Knockdown of FDL242 expression significantly enhanced the toxicity of trifluralin to peach aphid nymphs. Furthermore, the combined application of S-(+)-tebufenozide and trifluralin also significantly enhanced the toxicity of trifluralin to larvae.

[0114] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of inhibiting farnesol dehydrogenase expression in enhancing the toxicity of trifluralin; The farnesol dehydrogenase includes any one of the following (1) to (2): (1) The amino acid sequence of Aedes aegypti farnesol dehydrogenase FDL22 is shown in SEQ ID NO.1; the nucleotide sequence encoding the protein of Aedes aegypti farnesol dehydrogenase FDL22 is shown in SEQ ID NO.2; (2) The amino acid sequence of the peach aphid farnesol dehydrogenase FDL242 is shown in SEQ ID NO.13; the nucleotide sequence encoding the peach aphid farnesol dehydrogenase FDL242 protein is shown in SEQ ID NO.14; When the farnesol dehydrogenase is (1), the toxicity of the enhanced trifluralin is to improve the killing efficiency of trifluralin against Aedes aegypti. When the farnesol dehydrogenase is (2), the toxicity of the enhanced trifluralin is to improve the killing efficiency of trifluralin against peach aphids.

2. The application according to claim 1, characterized in that, Methods to inhibit the expression of farnesol dehydrogenase include: (A) knocking down farnesol dehydrogenase via dsRNA; (B) The expression of farnesol dehydrogenase was inhibited by knocking down farnesol dehydrogenase with dsRNA and by topical application of a juvenile hormone analogue, wherein the juvenile hormone analogue was S-(+)-tebufenozide.

3. A dsRNA of farnesol dehydrogenase FDL22 from Aedes aegypti mosquitoes, characterized in that, The nucleotide sequence of the dsRNA is shown in SEQ ID NO.

3.

4. A dsRNA of peach aphid farnesol dehydrogenase FDL242, characterized in that, The nucleotide sequence of the dsRNA is shown in SEQ ID NO.

15.

5. A synergistic insecticide, characterized in that, These include dsRNAs that inhibit farnesol dehydrogenase expression and trifluralin; the dsRNAs that inhibit farnesol dehydrogenase expression include the dsRNA of Aedes aegypti farnesol dehydrogenase FDL22. The nucleotide sequence of the dsRNA of the farnesol dehydrogenase FDL22 of Aedes aegypti is shown in SEQ ID NO.

3.

6. A synergistic insecticide, characterized in that, The invention includes dsRNA that inhibits farnesol dehydrogenase expression and trifluralin; the dsRNA that inhibits farnesol dehydrogenase expression includes the dsRNA of peach aphid farnesol dehydrogenase FDL242; the nucleotide sequence of the dsRNA of peach aphid farnesol dehydrogenase FDL242 is shown in SEQ ID NO.

15.

7. The application of the dsRNA of claim 3 or the synergistic insecticide of claim 5 in the control of pests; wherein the pest is Aedes aegypti.

8. The application of the dsRNA of claim 4 or the synergistic insecticide of claim 6 in the control of pests; wherein the pest is the peach aphid.

9. A method for controlling pests with the synergistic insecticide of claim 5, characterized in that, include: When the synergistic insecticide includes dsRNA that inhibits farnesol dehydrogenase expression, the dsRNA is delivered to the pest, and then the pest is exposed to trifluralin. The pest in question is the Aedes aegypti mosquito.

10. A method for controlling pests with the synergistic insecticide of claim 6, characterized in that, include: When the synergistic insecticide includes dsRNA that inhibits farnesol dehydrogenase expression, the dsRNA is delivered into the pest, and then the pest is exposed to trifluralin. The pest in question is the peach aphid.

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