Use of a dsrna in the control of bemisia tabaci resistant to flucycloxuron

Abstract

This invention provides the application of dsRNA in the control of fluoxazolamide-resistant whiteflies, belonging to the field of pesticide technology. The dsRNA is a dsRNA... BtUGT352B2 or ds BtUGT352F1 This invention discovered and confirmed two UGT genes. BtUGT352B2 , BtUGT352F1 The resistance of whiteflies to fluoxazolamide is closely related to the development of resistance through feeding specific dsRNA (dsRNA). BtUGT352B2 or ds BtUGT352F1 )inhibition BtUGT352B2 or BtUGT352F1 The expression of this substance can significantly reduce the resistance level of whiteflies to fluoxazolamide, and this discovery provides an important basis for developing new control strategies against fluoxazolamide-resistant whiteflies.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically the application of dsRNA in the control of fluoxazolamide-resistant whiteflies. Background Technology

[0002] Whiteflies ( Bemisia tabaci The tobacco whitefly, belonging to the order Hemiptera and family Aleyrodidae, originated in tropical and subtropical regions and has spread widely and caused widespread damage worldwide, making it one of the most destructive pests in the world. The whitefly severely impacts crop growth by sucking plant sap, transmitting plant viruses, and secreting honeydew that induces enzyme-induced diseases. In 2007, Nissan Corporation developed the first commercially available GABACls (GABA-gated Chloride channels) allosteric regulator insecticide—fluxametamide (FXM). This type of insecticide regulates the allosteric activity of insect GABACls, possessing a complex structure, high selectivity, and a novel mechanism of action. It exhibits no cross-resistance with GABACls antagonists, preventing the GABA (γ-aminobutyric acid) receptor from generating normal inhibitory postsynaptic sites, thus leading to insect death. It shows great promise for future applications. However, the long-term continuous use of chemical agents has led to whiteflies in the field developing resistance to fluoxazolamide, and the control effect has declined year by year; with the increase in the frequency of pesticide application, it is becoming increasingly difficult to control fluoxazolamide-resistant whitefly populations.

[0003] UDP-glycosyltransferases (UGTs) are a multifunctional superfamily of enzymes found in all organisms in nature (including plants, animals, bacteria, and fungi). Insect UGTs typically use UDP-glucose (UDPG) as a glycosyl donor, transferring glucose to the substrate through glycosylation reactions, promoting the metabolism and excretion of exogenous and endogenous compounds, thereby exerting a detoxification function. On the one hand, UGTs can prevent the disruption of internal systems by exogenous substances; on the other hand, UGTs can effectively regulate the internal environmental balance, thus playing a crucial role in the growth and development of plants and animals. Recent studies have found that the glycosylation of exogenous compounds catalyzed by insect UGTs plays an important role in the insect detoxification metabolic system. Insects can cope with the toxicological effects of pesticides by regulating the expression abundance of their own UGT genes. Currently, UGTs are known to participate in the development of resistance in insects to organophosphates, pyrethroids, neonicotinoids, and diamides.

[0004] This invention aims to explore the function of UGT in fluoxazolamide metabolism and resistance, in order to develop a novel control strategy for fluoxazolamide-resistant whitefly populations. Summary of the Invention

[0005] This invention provides an application of dsRNA in the control of fluoxazolamide-resistant whiteflies, providing important application basis for the development of new control strategies against fluoxazolamide-resistant whiteflies.

[0006] The technical solution of this invention is as follows: The application of a dsRNA in the control of fluoxazolamide-resistant whiteflies, wherein the dsRNA is a dsRNA. BtUGT352B2 or ds BtUGT352F1 , where ds BtUGT352B2 The nucleotide sequence is shown in SEQ ID NO.1, ds BtUGT352F1 The nucleotide sequence is shown in SEQ ID NO.2.

[0007] Preferably, the application method is to feed whiteflies ds BtUGT352B2 or ds BtUGT352F1 Inhibits the body of whiteflies BtUGT352B2 or BtUGT352F1 The expression of [something] reduces the resistance of whiteflies to fluoxazolamide.

[0008] Preferably, the feeding ds BtUGT352B2 or ds BtUGT352F1 The method is: using ds BtUGT352B2 or ds BtUGT352F1 The solution was used to feed the whiteflies.

[0009] Preferably, the solution contains ds BtUGT352B2 or ds BtUGT352F1 Sugar water.

[0010] More preferably, the concentration of sucrose in the sucrose water is 0.2 mg / L.

[0011] Preferably, the ds BtUGT352B2 or ds BtUGT352F1 The concentration in sucrose solution is 200~500 ng / µL.

[0012] Gene BtUGT352B2 or BtUGT352F1 Its application is in the control of fluoxazolamide-resistant whiteflies. BtUGT352B2 , BtUGT352F1 The nucleotide sequences are shown in SEQ ID NO. 3~4, respectively; the application method is: by reducing... BtUGT352B2 or BtUGT352F1 The expression level in whiteflies reduces their resistance to fluoxazolamide.

[0013] Beneficial effects: This invention discovered and confirmed two UGT genes. BtUGT352B2 , BtUGT352F1The resistance of whiteflies to fluoxazolamide is closely related to the development of resistance through feeding specific dsRNA (dsRNA). BtUGT352B2 or ds BtUGT352F1 )inhibition BtUGT352B2 or BtUGT352F1 The expression of this substance can significantly reduce the resistance level of whiteflies to fluoxazolamide, and this discovery provides an important basis for developing new control strategies against fluoxazolamide-resistant whiteflies. Attached Figure Description

[0014] Figure 1 To locate chromosomal regions of fluoxazolamide resistance-related genes using G' values ​​based on grouped pooled analysis of segregating populations (BSA) and the FST method; Figure 2 For target genes BtUGT352B2 , BtUGT352F1 , BtCPR , BtCYP415A1 Expression levels in the resistant population FMX-R and the susceptible population FMX-S whiteflies; Figure 3 The enzyme activity of UGT in whiteflies of the resistant population FMX-R and the susceptible population FMX-S; Figure 4 After feeding dsRNA, the resistant population of FMX-R whiteflies... BtUGT352B2 and BtUGT352F1 The relative representation of the values, where Figure A is... BtUGT352B2 The relative expression situation, Figure B is BtUGT352F1 The relative expression situation; Figure 5 After feeding dsRNA, the resistant population of FMX-R whiteflies... BtUGT352N1 and BtUGT352P1 The relative representation of the values, where Figure A is... BtUGT352N1 The relative expression situation, Figure B is BtUGT352P1 The relative expression of . Detailed Implementation

[0015] The present invention will now be described in detail with reference to specific embodiments.

[0016] Explanation of the source of experimental materials: Fluoxazolamide (99.7% technical grade): Nissan Chemical Industries, Ltd., Tokyo, Japan; TRIzol: Thermo Fisher Scientific, Inc., USA; Fluorescent dye SYBR Green Pro Taq HS: Aikerui Biotechnology; TranscriptAid T7 High-Yield Transcription Kit: Thermo Fisher Scientific, Inc.

[0017] The relevant information of the whitefly populations involved in the embodiments is shown in Table 1: Table 1. Whitefly Population Information

[0018] Example 1: Group-Based Analysis (BSA) of Separate Populations I. Constructing a Mixed Pool Male whiteflies from the FXM-R population were crossed with female whiteflies from the FXM-S population to produce the F1 generation. Females from the F1 generation were selected for parthenogenesis to produce the F2 generation. Adults from the F2 generation were screened, and individuals that survived treatment with 8000 mg / L fluoxazolamide for 24 hours were designated as the extremely resistant pool (R-pool), while individuals that showed severe poisoning symptoms after treatment with 2 mg / L fluoxazolamide for 24 hours were designated as the extremely sensitive pool (S-pool).

[0019] II. DNA / RNA Extraction and cDNA Synthesis 120 adult whiteflies were collected from each of the resistant population FMX-R, the susceptible population FMX-S, the extremely resistant mixed pool (R-pool), and the extremely susceptible mixed pool (S-pool). DNA was extracted using the CTAB method, incubated with RNase A, and precipitated with isopropanol to obtain the genomic DNA of the whitefly. The genomic DNA was used to construct a library and sequenced on the Illumina HiSeq 2500 platform (Zhenyue Biotechnology, Wuhan). The average depth of sequencing covering the entire genome was approximately 15×.

[0020] Fifty adult worms each from the resistant population FMX-R, the sensitive population FMX-S, the extremely resistant mixed pool (R-pool), and the extremely sensitive mixed pool (S-pool) were collected. After thorough grinding with liquid nitrogen, RNA was extracted using TRIzol and digested with DNase I to obtain total RNA. First-strand cDNA was synthesized from the total RNA using the PrimeScript RT reagent Kit with a gDNA eraser kit (Takara).

[0021] III. BSA Two complementary statistical methods (G' value and FST) were used for grouped pooled analysis (BSA) of segregating populations. SNP identification and preliminary screening of sequencing data were performed using GATK's Haplotyper method, with all screening parameters set according to GATK software recommendations. After variant detection, the SnpEff program was used for annotation analysis of the identified SNPs.

[0022] The results are as follows Figure 1 As shown. By Figure 1A significant consensus peak was identified in the 8.64-16.82 Mb region of chromosome 5, which contains four enzymes, including two UDP-glycosyltransferases (UGT352B2 and UGT352F1), one cytochrome P450 reductase (CPR), and one cytochrome P450 monooxygenase (CYP415A1). These four enzymes may be associated with resistance of whiteflies to fluoxazolamide.

[0023] Based on the SNP site screening in the BSA results, no population-specific SNP sites were found between the four enzyme-encoded genes in the resistant whitefly population FMX-R and the susceptible whitefly population FMX-S. Therefore, the expression levels of the four enzyme-encoded genes in the resistant and susceptible whitefly populations were subsequently tested.

[0024] Example 2: RT-qPCR I. Screening of Target Gene and Internal Reference Gene The complete sequence information of the encoding genes of the above four enzymes, UGT352B2, UGT352F1, CPR, CYP415A1, and Rdl (Resistant to dieldrin), was found through the whitefly genomic DNA database. Then, specific primers for RT-qPCR detection were designed using Primer 6.0 software. In order to eliminate the influence of pesticide treatment on the internal reference genes, succinate dehydrogenase complex subunit A (SDHA) and heat shock protein 40 (HSP40) were selected as dual internal reference genes to correct the experimental data.

[0025] II. Total RNA Extraction and Reverse Transcription Total RNA was extracted from the resistant population FMX-R and the susceptible population FMX-S whiteflies using the TRIzol method. 1000 ng of qualified RNA was used as a template, and the first strand of cDNA was synthesized using the Takara PrimeScript RTreagent Kit with gDNA Eraser reverse transcription kit, which has the function of removing genomic DNA.

[0026] III. RT-qPCR The RT-qPCR reaction system was configured according to the instructions for the fluorescent dye SYBR Green Pro Taq HS, and three technical replicates were set up for the detection of each gene.

[0027] The RT-qPCR reaction system is as follows: SYBR Green Pro Taq HS 10μL, upstream / downstream primers 1μL, cDNA template 1μL, and enzyme-free water 7μL; The RT-qPCR reaction conditions were as follows: 95℃, 30s; 95℃, 5s; 60℃, 30s; 40 cycles. Using 2... -ΔΔCt Method for targeting genes BtUGT352B2 , BtUGT352F1 , BtCPR , BtCYP415A1 as well as BtRdl The relative expression levels were calculated and analyzed.

[0028] The results are as follows Figure 2 As shown. By Figure 2 It can be seen that there are 2 UGT genes ( BtUGT352B2 and BtUGT352F1 Expression was significantly upregulated in the resistant population FMX-R, with levels increasing by 5.78-fold and 4.96-fold, respectively, compared to the sensitive population FMX-S; however, CPR , CYP415A1 There were no significant differences between resistant and susceptible populations.

[0029] Example 3: Effect of UGT inhibitors on fluoxazolamide resistance in whiteflies First, the median lethal concentration (LC50) of fluoxazolamide insecticide against whitefly populations was determined using the agar moist leaf immersion method. 50 The specific procedures are as follows: Fluoxazolamide was prepared to the highest concentration for the bioassay experiment using deionized water containing 0.05% Triton X-100. This was then serially diluted to obtain five concentrations, with deionized water containing 0.05% Triton X-100 serving as a control. Cotton leaf discs were sequentially immersed in the pesticide and then placed in a bioassay device containing 1% agar. Three-day-old whiteflies were placed in each bioassay device. After 48 hours, the whitefly mortality rate was recorded. The concentration and mortality rates were input into PoloPlus software for data analysis to obtain the LC50. 50 and its 95% confidence interval.

[0030] Then, the effect of the UGT inhibitor sulfinpyrazone on the resistance level of fluoxazolamide in different whitefly populations (FMX-R, FMX-S) was determined: sulfinpyrazone (Sul) was dissolved in acetone to obtain a sulfinpyrazone solution with a concentration of 100 mg / L; fluoxazolamide was serially diluted with the sulfinpyrazone solution, and then its effect on the resistance level of different whitefly populations was determined according to the above-described agar moist leaf soaking method. The results are shown in Table 2: Table 2. Toxicity of fluoxazolamide to different whitefly populations and synergistic effect of sulfinpyrazone on fluoxazolamide.

[0031] As shown in Table 2, the synergistic effect of the UGT inhibitor sulfinpyrazone on the FXM-S whitefly population was 1.3 times, and the synergistic effect on the FXM-R whitefly population was 2.9 times. This suggests that UGT may be involved in the resistance of whiteflies to fluoxazolamide.

[0032] Example 4: Determination of UGT enzyme activity in different whitefly populations Based on the function of UGT in catalyzing the conversion of α-naphthol to α-naphthyl glucoside, the UGT enzyme activity of whiteflies was determined. The specific operation was as follows: 40 adult whiteflies were collected and ground with 0.05 M, pH 7.8 PBS buffer. The mixture was centrifuged at 12000 g for 15 min at 4 ℃, and the supernatant (enzyme solution) was collected. Magnesium chloride (10 mM), UDP-glucose (UDPG) (1 mM), and the enzyme solution were added sequentially to a 1.5 mL centrifuge tube. Distilled water was added to a final concentration of 960 μL, and the mixture was incubated in a 37 ℃ water bath to obtain a mixed system. The mixed system was aliquoted into microplates, with 240 µL added to each well. Finally, 10 µL of 200 µM UDP-glucose was added. α-Naphthol was thoroughly mixed; the enzyme-linked immunosorbent assay (ELISA) reader was quickly started, and α-naphthyl glucoside was continuously monitored for 10 min at an excitation wavelength of 287 nm and an emission wavelength of 335 nm, with a slit width of 5 nm. The amount of product generated was calculated based on the standard curve of α-naphthyl glucoside, and the enzyme activity of UGT was calculated based on the protein content. Three biological replicates were set up for each population.

[0033] The enzyme activity assay results of UGT in the resistant population FMX-R and the susceptible population FMX-S whiteflies are as follows: Figure 3 As shown. By Figure 3 The results showed that the UGT enzyme activity in the fluoxazolamide-resistant whitefly population (FXM-R) was increased by 2.2 times compared to the susceptible population (FXM-S). These results suggest that the increased UGT enzyme activity is likely involved in the whitefly's resistance to fluoxazolamide.

[0034] Example 5: RNA interference (RNAi) I. Synthesis of dsRNA 2 UGT genes BtUGT352B2 and BtUGT352F1 The nucleotide sequences are shown in SEQ ID NO.3~4, respectively.

[0035] Total RNA was extracted from the fluoxazol-resistant population (FXM-R) of whiteflies using Trizol reagent and PrimeScript was used. TM II. First-strand cDNA was synthesized using the 1st Strand cDNA Synthesis Kit; using the cDNA as a template, it was amplified by PCR using Apex HF HS DNA Polymerase. BtUGT352B2and BtUGT352F1 The gene fragments were then used to synthesize dsRNAs using the TranscriptAid T7 high-yield transcription kit. These dsRNAs were ds... BtUGT352B2 and ds BtUGT352F1 The nucleotide sequences are shown in SEQ ID NO.1~2, respectively.

[0036] Among them, amplification BtUGT352B2 The primer sequences used for the gene fragments are shown in SEQ ID NO. 5-6, respectively; amplification BtUGT352F1 The primer sequences used for the gene fragments are shown in SEQ ID NO.7~8.

[0037] The above dsRNA synthesis system is as follows: 5×TranscriptAid Reaction Buffer 5μL, ribonucleotides (A / G / C / U, 100mM) 2μL each, DNA template 2μg, enzyme-free water to make up to 20μL, mix thoroughly, and incubate at 37℃ for 4h.

[0038] Simultaneously, dsRNA (ds) synthesizes green fluorescent protein (GFP). GFP () was used as a control group for subsequent RNA interference (RNAi) experiments.

[0039] II. RNAi ds BtUGT352B2 ds BtUGT352F1 and ds GFP The dsRNAs were dissolved separately in 0.2 mg / L sucrose water to achieve a final concentration of 250 ng / µL, yielding feeding solutions. These solutions were then added to feeding devices, which were 50 mL centrifuge tubes sealed with insect-proof netting around the sides and bottom, and sealed with a centrifuge tube cap. The feeding solutions were then separated from over 350 three-day-old fluoxazolamide-resistant (FXM-R) adult whiteflies using a polytetrafluoroethylene (PTFE) membrane. The whiteflies' mouthparts could pierce the PTFE membrane, allowing them to extract the feeding solution from the inside. After 72 hours of feeding, surviving whiteflies were collected for subsequent qPCR and bioassay experiments.

[0040] III. RT-qPCR Total RNA was extracted from surviving whiteflies after RNAi using Trizol reagent. Then, mRNA was reverse transcribed using the PrimeScript™ RTreagent Kit with gDNA eraser, following the kit instructions, to synthesize first-strand cDNA. The cDNA was used as a template for RT-qPCR, with each treatment performed in triplicate. Two PCR reactions were then conducted. -ΔΔCt Relative quantitative analysis of whiteflies after RNAiBtUGT352B2 , BtUGT352F1 The relative expression of .

[0041] The results are as follows Figure 4 As shown. By Figure 4 It can be seen that, compared with feeding ds GFP Compared with the control group, ds BtUGT352B2 ds BtUGT352F1 After feeding fluoxazolamide-resistant whiteflies for 72 hours, the expression levels of the corresponding genes were significantly reduced, among which, BtUGT352B2 The expression level was 0.32 times that of the control group. BtUGT352F1 The expression level was 0.46 times that of the control group. These results indicate that feeding ds... BtUGT352B2 ds BtUGT352F1 Both can effectively knock down the levels of whiteflies. BtUGT352B2 , BtUGT352F1 The level of expression.

[0042] IV. Effects of RNAi on fluoxazolamide resistance in whiteflies The effect of RNAi on fluoxazolamide resistance in whiteflies was determined using the same agar-soaked leaf-immersion method as in Example 3: Fluoxazolamide was prepared to the highest concentration for the bioassay experiment using deionized water containing 0.05% Triton X-100, and then serially diluted to obtain five concentrations. Deionized water containing 0.05% Triton X-100 was used as a control (natural mortality rate was measured). Cotton leaf discs were sequentially immersed in the drug and then attached to bioassay devices containing 1% agar. Adult whiteflies that had undergone RNAi were placed in each bioassay device, and three biological replicates were set for each concentration. All centrifuge tubes were cultured under standard laboratory conditions, and mortality data were recorded after 48 hours. The LC50 was calculated using PoloPlus software. 50 The values ​​and their 95% confidence intervals. It should be noted that during the above bioassays, ds were fed... GFP The natural mortality rate of whiteflies after 48 hours was 1.82 ± 4.07%, and the feeding dose was ds BtUGT352B2 and ds BtUGT352F1 The natural mortality rates of whiteflies after 48 hours were 2.00±4.47% and 1.67±3.73%, respectively, with no significant difference in natural mortality rates among different feeding groups.

[0043] The results are shown in Table 3: Table 3. Feeding ds BtUGT352B2 ds BtUGT352F1 Post-FXM-R population toxicity regression curves for fluoxazolamide

[0044] Table 3 shows that the levels of interference-resistant population (FXM-R) within the population are...BtUGT352B2 or BtUGT352F1 Following gene knockdown, the resistance level of whitefly resistant populations to fluoxazolamide was significantly reduced. Specifically, knockdown... BtUGT352B2 Post-LC 50 The value decreased by 2.6 times; knockdown BtUGT352F1 Post-LC 50 The value decreased by 2.8 times; due to ds BtUGT352B2, ds BtUGT352F1 Groups and ds GFP Group LC 50 The 95% confidence intervals did not overlap, indicating that the LC50 of the resistant population (FXM-R) to fluoxazolamide was not significantly different. 50 Significantly reduced.

[0045] Comparative Example 1: BtUGT352B2 and BtUGT352F1 Specificity in whitefly resistance to fluoxazolamide I. Synthesis of dsRNA To verify the two UGT genes BtUGT352B2 and BtUGT352F1 To investigate whether there is specificity in the resistance of whiteflies to fluoxazolamide, two other UGT genes of the BtUGT352 family were amplified by PCR using the cDNA synthesized in Example 5 as a template. BtUGT352N1 (SEQ ID NO.9) and BtUGT352P1 The gene fragment of (SEQ ID NO.10) was then used to synthesize dsRNA using the TranscriptAid T7 high-yield transcription kit. The dsRNAs were ds BtUGT352N1 and ds BtUGT352P1 The nucleotide sequences are shown in SEQ ID NO.11~12, respectively.

[0046] Among them, amplification BtUGT352N1 The primer sequences used for the gene fragment are shown in SEQ ID NO. 13-14, respectively; amplification BtUGT352P1 The primer sequences used for the gene fragments are shown in SEQ ID NO.15~16, respectively.

[0047] II. RNAi Following the same method as in Example 5, using ds BtUGT352N1, ds BtUGT352P1 and ds GFP RNAi was performed on adult whiteflies of the fluoxazolamide-resistant population (FXM-R). After feeding for 72 hours, the surviving whiteflies were collected for subsequent qPCR and bioassay experiments, using the same methods as in Example 5.

[0048] III. RT-qPCR After RNAiBtUGT352N1, BtUGT352P1 The relative expression situation is as follows Figure 5 As shown. By Figure 5 It can be seen that, compared with feeding ds GFP Compared with the control group, ds BtUGT352N1, ds BtUGT352P1 After feeding fluoxazolamide-resistant whiteflies for 72 hours, the expression levels of the corresponding genes were significantly reduced, among which, BtUGT352N1 The expression level was 0.23 times that of the control group. BtUGT352P1 The expression level was 0.37 times that of the control group. These results indicate that feeding ds... BtUGT352N1ぁ ds BtUGT352P1 Both can effectively knock down the levels of whiteflies. BtUGT352N1, BtUGT352P1 The level of expression.

[0049] IV. Effects of RNAi on fluoxazolamide resistance in whiteflies The effect of RNAi on fluoxazolamide resistance in whiteflies was determined by the agar-soaked leaf immersion method. The results are shown in Table 4. Table 4. Feeding ds BtUGT352N1ぁ ds BtUGT352P1 Post-FXM-R population toxicity regression curves for fluoxazolamide

[0050] Table 4 shows that the levels of interference-resistant population (FXM-R) within the population are... BtUGT352N1 or BtUGT352P1 Following gene sequencing, the resistance level of whitefly resistant populations to fluoxazolamide did not change significantly. Specifically, knocking down... BtUGT352N1 Post-LC 50 The value was 183.63 mg / L, knockdown BtUGT352P1 Post-LC 50 The value was 179.09 mg / L, which was not significantly different from the control group (184.51 mg / L); and the LC values ​​of the three groups were also not significantly different. 50 The 95% confidence intervals highly overlap, indicating that the knockdown... BtUGT352N1 or BtUGT352P1 It does not reduce the resistance of whiteflies to fluoxazolamide.

[0051] The above results indicate that BtUGT352B2 or BtUGT352F1 The expression level of this gene significantly affects the survival ability of whiteflies in fluoxazolamide environments and is a key gene determining the development of fluoxazolamide resistance in whiteflies; when fed ds BtUGT352B2 or ds BtUGT352F1 Afterwards, all of them were able to significantly reduce the resistance level of whiteflies to fluoxazolamide.

Claims

1. The application of dsRNA in the control of fluoxazolamide-resistant whiteflies, characterized in that, The dsRNA is ds BtUGT352B2 or ds BtUGT352F1 , where ds BtUGT352B2 The nucleotide sequence is shown in SEQ ID NO.1, ds BtUGT352F1 The nucleotide sequence is shown in SEQ ID NO.

2.

2. The application as described in claim 1, characterized in that, The application method involves feeding whiteflies ds BtUGT352B2 or ds BtUGT352F1 Inhibits the body of whiteflies BtUGT352B2 or BtUGT352F1 The expression of [something] reduces the resistance of whiteflies to fluoxazolamide.

3. The application as described in claim 2, characterized in that, The feeding ds BtUGT352B2 or ds BtUGT352F1 The method is: using ds BtUGT352B2 or ds BtUGT352F1 The solution was used to feed the whiteflies.

4. The application as described in claim 3, characterized in that, The solution contains ds BtUGT352B2 or ds BtUGT352F1 Sugar water.

5. The application as described in claim 4, characterized in that, The concentration of sucrose in the sucrose solution is 0.2 mg / L.

6. The application as described in claim 4, characterized in that, The ds BtUGT352B2 or ds BtUGT352F1 The concentration in sucrose solution is 200~500 ng / µL.

7. Genes BtUGT352B2 or BtUGT352F1 The application is characterized by, Used to control fluoxazolamide-resistant whiteflies, BtUGT352B2 , BtUGT352F1 The nucleotide sequences are shown in SEQ ID NO. 3~4, respectively; the application method is: by reducing... BtUGT352B2 or BtUGT352F1 The expression level in whiteflies reduces their resistance to fluoxazolamide.

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