Brown planthopper nllipase 1 gene, coding protein and application thereof

Abstract

The application discloses a Nilaparvata lugens Nllipase1 gene, a coded protein and application thereof. The Nilaparvata lugens Nllipase1 gene, a nucleotide sequence of the Nllipase1 gene is shown as SEQ ID NO. 1. The application obtains a cDNA sequence of the Nilaparvata lugens Nllipase1 gene, adopts RNAi technology of microinjection dsRNA, silences the Nilaparvata lugens Nllipase1 gene, causes the Nilaparvata lugens to produce a feeding rejection effect; obviously, the dsRNA of the gene has important application value in the field of Nilaparvata lugens prevention.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to the brown planthopper. Nllipase1 Applications of the gene, its dsRNA, and RNA interference targeting the gene in the control of brown planthoppers. Background Technology

[0002] Brown planthopper ( Nilaparvata lugens The brown planthopper (Stål.), belonging to the family Delphacidae (Hemiptera) (Wan et al., 2019), is one of the most threatening pests of rice crops in tropical Asia and southern China (Bao et al., 2019; 2014). A typical monophagous pest, the brown planthopper is characterized by its explosive outbreaks, migratory nature, concealment, and destructiveness. It sucks sap from the phloem of rice plants, causing leaf withering. It also lays eggs on rice stems, leading to yellowing and lodging of the plants, and can indirectly transmit plant viruses (Xue et al., 2014; Xu et al., 2019). The brown planthopper has an extremely high reproductive rate; each female can lay 500-1000 eggs under suitable conditions, and it reproduces several generations annually. The number of brown planthoppers, determined by its reproductive capacity, is the most direct factor affecting rice damage (Sogawa, 2003; Yu, 2007). Brown planthoppers, which feed on sap, secrete large amounts of honeydew, which contains abundant amino acids and sugars, making rice susceptible to various bacterial infections. Large-scale outbreaks of brown planthoppers can lead to a significant decrease in rice yield, and in severe cases, complete crop failure (Li Chunfeng, 2019). Currently, insecticides are commonly used to control brown planthoppers; however, the extensive use of these pesticides has led to resistance in brown planthoppers, such as tocarbamate, pyrethroids, and even synthetic pyrethroids (Nagata, 1982; Lakshmi et al., 2010). Furthermore, the use of insecticides severely disrupts the ecological balance. Therefore, actively developing and applying biological control technologies for brown planthoppers is of great significance for the scientific and effective control of brown planthopper damage in my country and worldwide, and for ensuring my country's food security and environmental safety.

[0003] RNA interference (RNAi) is a technique that uses double-stranded RNA (dsRNA) to introduce into host cells, silencing related genes and thus interfering with host growth and development. It is widely used in the biological control of insects. RNAi technology has advantages such as specificity, broad spectrum, high efficiency, systematic approach, and environmental safety (Whitten, 2019). Since Montgomery (1998) first observed RNA interference in *Caenorhabditis elegans* (…), it has been widely applied in the biological control of insects. Caenorhabditis elegansSince its discovery in insects, RNA interference (RNAi) technology has become a powerful tool for knocking down or eliminating genes, widely used in basic biology and pest control research (Montgomery et al., 1998; Monte, 2023). Microinjection is one of the most commonly used methods for RNA interference (Ober and Jockusch, 2006). Microinjection involves injecting a certain amount of in vitro synthesized dsRNA into the target insect to cause RNA interference. In recent years, many researchers have used microinjection technology to achieve RNAi in insects, with most studies focusing on the formation of insect eggs and embryonic development (McFarlane et al., 2021). Microinjection has advantages such as high accuracy, stable RNAi efficiency, no limit on the size of the injected gene, no need for vectors, and no special requirements for the target organism. It has been applied to various aspects of insect genetics research, especially its ability to efficiently screen for target genes required for RNAi biocontrol, laying a solid foundation for subsequent pest biocontrol.

[0004] With the widespread dissemination and practice of environmental protection and sustainable development strategies, the control of pests can no longer rely on traditional chemical pesticides or fungicides (Chang Tong et al., 2023). Zhang Jianzhen et al. (2021) categorized pest control into feeding methods, transgenic methods, and nanocarrier-mediated methods based on different dsRNA delivery methods. With the rapid development of nanotechnology in recent years, nanomaterials have been widely used as a novel biological carrier in the biological control of pests (Bi et al., 2023). Lin et al. (2017) used the German cockroach... Blattella germanica Continuous ingestion of dsRNA lipid complexes demonstrated that dsRNA encapsulated by cationic liposomes was sufficient to slow down midgut dsRNA degradation and increase mortality in German cockroaches by significantly inhibiting tubulin expression. Zhang et al. (2019) used chitosan to deliver dsRNA of the chitin synthase 2 gene orally, achieving a gene knockout efficiency of 40%-60%, leading to increased mortality in *Amblyseius gambiae*. Castellanos et al. (2019) used liposomes and EDTA to formulate dsRNA targeting key genes, increasing the mortality rate of male *Bryophyte spp.*, a neotropical bed bug, induced by oral RNAi. Yuan et al. (2023) used nanocarriers to deliver dsRNA... NlsNPF significantly increased the mortality rate of brown planthoppers. As a novel insect control method, RNAi biocontrol technology still faces many challenges. Successfully introducing dsRNA into the host and enabling it to be expressed is a key step in this technology. However, factors such as the insect gut environment and enzyme interference, the amount of dsRNA introduced, and the dsRNA introduction time can interfere with this stage and ultimately affect the effectiveness of RNAi application (Li Qiang et al., 2021). Summary of the Invention

[0005] The purpose of this invention is to provide a clone from the brown planthopper. Nllipase1 cDNA and protein sequences of the gene. Silencing of the brown planthopper using RNAi mediated by microinjection and adjuvants. Nllipase1 Genes that reduce the survival rate of brown planthoppers. This invention is achieved through the following technical solution:

[0006] This invention provides a brown planthopper Nllipase1 Genes, the ones mentioned Nllipase1 The nucleotide sequence of the gene's cDNA is shown in SEQ ID NO.1.

[0007] The present invention also provides a brown planthopper Nllipase1 The protein encoded by the gene has the amino acid sequence shown in SEQ ID NO.2.

[0008] This invention also designs a dsRNA for the brown planthopper gene, specifically a sequence shown in SEQ ID NO.1 that can achieve inhibition. Nllipase1 A double-stranded RNA composed of nucleotides that affect gene expression and their inverse complementary nucleotide sequences.

[0009] Brown planthopper Nllipase1 Using genes as templates, ds Nllipase1 -F, ds Nllipase1 -R is used as a primer for PCR amplification to obtain the PCR amplification product, and then dsRNA is synthesized.

[0010] ds Nllipase1 -F:

[0011] 5'- GGATCCTAATACgACTCACTATAGG ccgatggattaggttttgtact-3'

[0012] ds Nllipase1 -R:

[0013] 5'- GGATCCTAATACGACTCACTATAGG acgattttggatcacaatctttcttg-3'.

[0014] This invention also protects the use of the above-mentioned dsRNA in the preparation of products, the uses of which are any one of the following (1)-(3):

[0015] 1) Controlling brown planthoppers or preparing products for controlling brown planthoppers;

[0016] 2) Reduce the survival rate of brown planthoppers or prepare products that reduce the survival rate of brown planthoppers;

[0017] 3) Suppressing brown planthopper genes Nllipase1 Gene expression or preparation to inhibit brown planthoppers Nllipase1 Products that express genes.

[0018] This invention also provides a method for controlling brown planthoppers, which involves introducing the aforementioned dsRNA into brown planthoppers to control them, reduce their survival rate, or inhibit their growth. Nllipase1 Gene expression.

[0019] Preferably, the introduction method is microinjection, in which dsRNA is injected into the brown planthopper, and then the brown planthopper feeds on it. Results showed that both the amount of food consumed and the survival rate of the rice planthopper decreased.

[0020] The present invention has the following positive and beneficial technical effects:

[0021] Experiments have shown that this invention yields brown planthoppers. Nllipase1 The cDNA sequence of the gene was used to silence the brown planthopper using RNAi technology with microinjection of dsRNA. Nllipase1 The gene causes the brown planthopper to exhibit a feeding refusal effect; clearly, the dsRNA of this gene has important application value in the field of brown planthopper control. Attached Figure Description

[0022] Figure 1 The expression patterns of brown planthoppers in different tissues and at different ages.

[0023] A: In the fat body (Fb), midgut (Mg), ovary (Ov), epidermis (In), legs (Lg), and head (Hd) of the brown planthopper Nllipase1 Expression level.

[0024] B: Brown planthopper nymphs, 1st-5th instars; adults, 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, and 15th day old. Nllipase1 Expression level.

[0025] Figure 2 For microinjection Nllipase1 The relative expression level of genes.

[0026] Figure 3 Measurement of feeding holes of brown planthoppers.

[0027] Figure 4 The honeydew amount and acquired body weight of brown planthoppers feeding on rice under different treatments were investigated.

[0028] A: Honeydew levels in rice fed by brown planthoppers under different treatments

[0029] B: Acquired body weight of brown planthoppers feeding on rice under different treatments

[0030] Figure 5 Mortality rates of brown planthoppers under different treatments. Detailed Implementation

[0031] The following embodiments further illustrate the content of the present invention, but should not be construed as limiting the present invention. Any modifications or substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit and essence of the invention are within the scope of the present invention.

[0032] Unless otherwise specified, the techniques used in the embodiments are conventional techniques well known to those skilled in the art. Unless otherwise specified, all materials and reagents used in the following embodiments are commercially available.

[0033] The brown planthoppers used in the following examples were raised at the Institute of Plant Protection, Guangdong Academy of Agricultural Sciences. The rearing temperature was 26±2℃.

[0034] Example 1: Brown Planthopper Nllipase1 Cloning of genes

[0035] Total RNA was extracted from brown planthoppers and reverse-engineered into cDNA. Primers were designed based on this sequence, and the 5' and 3' end sequences of the candidate gene were obtained using the TaKaRa 5' and 3' Full RACE kit. The transcription start and termination sites of the candidate gene were determined, and the full-length cDNA sequence of the gene was assembled. Primers were then resynthesized based on the full-length cDNA sequence. Nllipase1 -F, Nllipase1 -R, amplification Nllipase1 The full-length cDNA was obtained, and the ORF was predicted. The ORF sequence is shown in SEQ ID NO.1 of the sequence listing, and the encoded amino acid sequence is shown in SEQ ID NO.2.

[0036] Nllipase1 -F:5'-atgtcgtctgattgcttcctc-3'

[0037] Nllipase1-R:5'-acgattttggatcacaatctttcttg-3'

[0038] Example 2

[0039] Total RNA was extracted from six different tissues (ovary, head, midgut, epidermis, fat body, and foot) of the brown planthopper using Trizol in an RNase-free environment. The RNA was reverse transcribed into cDNA and then detected using qPCR. Nllipase1 Expression levels in various tissues (detection primers see Q) Nllipase1 -F and Q Nllipase1-R ), the result is as follows Figure 1 As shown in A: Nllipase1 It is expressed in various tissues of the brown planthopper, with the highest expression level in the ovary, followed by the fat body and midgut.

[0040] Q Nllipase1 -F:5'-TGCCAAAGAGAACCTCAGCATG-3'

[0041] Q Nllipase1-R :5'-ACGATTTTGGATCACAAATCTTTCTTG-3'

[0042] Total RNA was extracted from brown planthoppers at different instars (1st-5th instar nymphs and short-winged female brown planthoppers at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, and 15 days after emergence), then reverse transcribed into cDNA, and detected using qPCR. The results are shown below. Figure 1 As shown in B: Quantitative results are displayed. Nllipase1 Expression was observed at various time points in the brown planthopper, including on the 5th day of adulthood. Nllipase1 The expression level is highest at this stage, and then gradually decreases thereafter.

[0043] Example 3

[0044] Used to silence brown planthoppers Nllipase1 dsRNA of genes (ds Nllipase1 ) and the green fluorescent protein (GFP) gene ds used as a control. GFP Preparation

[0045] 1. Using the cDNA obtained in Example 1 as a template, ds Nllipase1 -F, ds Nllipase1 -R is the primer used for PCR amplification to obtain the PCR amplification product.

[0046] ds Nllipase1 -F (forward primer):

[0047] 5'- GGATCCTAATACgACTCACTATAGG ccgatggattaggttttgtact-3'

[0048] ds Nllipase1 -R (reverse primer):

[0049] 5'- GGATCCTAATACGACTCACTATAGG acgattttggatcacaatctttcttg-3'

[0050] The underlined area is the T7 RNA polymerase promoter sequence.

[0051] 2. Using a GFP-containing plasmid already available in the laboratory as a template, ds GFP -F, ds GFP -R is the primer used for PCR amplification to obtain the PCR amplification product.

[0052] ds GFP -F (forward primer):

[0053] 5'- GGATCCTAATACGACTCACTATAGG TGACCAGGCTGACCTAC-3'

[0054] ds GFP -R (reverse primer):

[0055] 5'- GGATCCTAATACGACTCACTATAGG TTGATGCCGTTCTACTGG-3'

[0056] 3. The amplification product is recovered, A is added, and ligated into the pMD18-T (TAKARA) vector. Positive clones are sent for sequencing. Once a correct clone is obtained, it is amplified using the same primers as above, and the amplification product is purified and concentrated to a concentration of 1 μg / μL. This product serves as the template for dsRNA synthesis.

[0057] 4. Add the following proportions: 10 μL transcription buffer, 1 μL each of ATP, CTP, GTP, and UTP (100 mM), 1.25 μL RNase inhibitor, 1 μL T7 RNA Polymerase, 1.5 μg template, and DEPC H2O to a final volume of 49 μL. Gently swirl to mix and centrifuge briefly. Place in a PCR apparatus and program: 37°C, 4 h; 75°C, 5 min; store at 16°C. Aspirate 1 μL and perform gel electrophoresis. Once the target band is detected, proceed to the next step.

[0058] 5. Mix 6 μL of 10X Reaction buffer, 12 μL of DNase, 0.5 μL of RNase, and 1.5 μL of DEPC H2O thoroughly. Centrifuge briefly and place in a PCR instrument at 37°C for 30 min. Then, remove and add 1 μL of EDTA (Fementas), and return to the PCR instrument at 65°C for 5 min to terminate the reaction. Aspirate 1 μL, dilute 10-fold, and use 2 μL for gel electrophoresis. Measure the concentration of the remaining 2 μL using a Nanodrop UV spectrophotometer. If the detection band is a single, bright band, and the OD... 260 / 280 A value between 1.8 and 2.0 indicates good dsRNA quality, allowing for the next step: RNA phenol-chloroform extraction.

[0059] 6. Perform routine phenol-chloroform extraction to remove proteins. Adjust the dsRNA concentration to 5 μg / μL, aliquot into 10 μL tubes, and store at -80°C. This yields the silent brown planthopper. Nllipase1 dsRNA of genes (ds Nllipase1 ) and the green fluorescent protein (GFP) gene ds used as a control. GFP .

[0060] Example 4ds Nllipase1 and ds GFP Microinjection and effect testing

[0061] 1. Brown planthopper preparation: Take 30 females and 10 males and place them in a cup containing TN1 seedlings. After 24 hours, remove the adults. Let them hatch into adults for 1 day before injection.

[0062] 2. Plate preparation: Weigh 1.5g of agar powder and add it to 100mL of water, boil, pour into a glass petri dish, and let it solidify for later use.

[0063] 3. Injection: Place 5-8 similarly sized insects in a test tube and anesthetize them with CO2 for 20 seconds. Do not blow CO2 directly onto the insects to avoid collisions and injury. Then, place the insects on a 1.5% agar plate, abdomen facing upwards. Inject using a Nanoliter 2010 microinjector according to the instructions. The injection site is between the anterior and mesothorax of the brown planthopper. The injection volume is 46 μL (5 μg / μL).

[0064] 4. After injecting brown planthoppers with dsRNA, samples were taken starting 24 hours post-injection, with three planthoppers sampled every 24 hours for a total of 72 hours. Samples were also taken from both uninjected and injected planthoppers. GFP Brown planthoppers were used as a control, and RT-PCR was used to verify changes in gene expression levels.

[0065] The specific operation is as follows: After sampling, RNA was extracted (refer to Example 2). The RNA was reversed using the TAKARA PrimeScript RTreagent Kit with gDNA Eraser (catalog number RR047A) according to the instructions to obtain the reversed product. 5 μL of the reversed cDNA was diluted 10-fold and subjected to real-time quantitative PCR on a CFX96Touch™ Real-Time PCR Detection System (Bio-Rad) instrument, following the reaction system: DNase / RNase-Free ddH2O 2.9 μL, 2X Supermix 4 μL, Primers (5mM) 0.6 μL, cDNA template 0.5 μL. Reaction conditions: 95°C pre-denaturation for 2 min, 95°C denaturation for 5-10 s, TM (55-65°C) annealing extension for 30 s. The last two steps were repeated for 30 cycles. Finally, a melting curve was performed at 65°C-95°C, increasing by 0.5°C for 5 s each step, to determine the specificity of the amplified product. -△△Ct The method is to conduct data analysis.

[0066] Internal reference gene Nl Actin primers:

[0067] Q Nlactin -F:5'-TGCGTGACATCAAGGAGAAGC-3'

[0068] Q Nlactin -R:5'-CCATACCCAAGAAGGAAGGCT-3'

[0069] Nllipase1 Quantitative primers:

[0070] Q Nllipase1 -F:5'-TGCCAAAGAGAACCTCAGCATG-3'

[0071] QNllipase1-R: 5'-ACGATTTTGGATCACAAATCTTTCTTG-3'

[0072] 5. Use a microinjector to inject ds Nllipase1 Subsequently, at 24h, 48h, and 72h, three samples of two surviving brown planthoppers were randomly collected. Total RNA from the whole planthopper was extracted, reverse transcribed into cDNA, and used for quantitative real-time PCR detection. The results are as follows. Figure 2 As shown, microinjection ds Nllipase1 The experimental group, compared with uninjected brown planthoppers and those injected with ds GFP Compared with the control group, from 48 hours after injection, the brown planthopper's bodyNllipase1 The relative expression level of the gene was significantly reduced, showing a significant difference compared to the control. P <0.05). The results show that by microinjecting ds Nllipase1 Can cause internal infection in brown planthoppers Nllipase1 The RNAi effect of the gene led to a significant decrease in gene expression. Therefore, it can be concluded that this dsRNA fragment can successfully induce target gene silencing in brown planthoppers.

[0073] Example 5: Silence Nllipase1 Impact on feeding behavior of brown planthoppers

[0074] To further verify Nllipase1 To investigate the influence of genes on the feeding behavior of brown planthoppers, we statistically analyzed the number of feeding marks at fixed locations on brown planthoppers within 24 hours. First, we injected ds... Nllipase1 Brown planthoppers were placed on single rice plants to feed after 24 hours. After 24 hours, the planthoppers were removed, and the rice stems from which the planthoppers had fed were soaked in eosin Y for 12 hours. After 12 hours, the planthoppers were removed and observed under a stereomicroscope to examine the feeding marks. Results are as follows: Figure 3 As shown. ds Nllipase1 The number of feeding traces in the group compared to the control group and ds GFP The group increased significantly, indicating that ds Nllipase1 It has an inhibitory effect on feeding by brown planthoppers.

[0075] Example 6 Silence Nllipase1 Effects on feeding amount of brown planthopper

[0076] For piercing-sucking insects like the brown planthopper, its feeding amount is closely related to its damage output; to reflect its harmfulness, the amount of honeydew secreted is often used as a measure. [The injection...] Nllipase1 Three days after the gene's dsRNA was transferred, brown planthoppers migrated to rice plants to feed. The amount of honeydew secreted over 24 hours and the acquired body weight were recorded. Results are as follows: Figure 4 As shown in A, ds Nllipase1 The amount of honeydew excreted by the group was significantly lower than that of the control group and the ds group. GFP Group. Injection Nllipase1 Brown planthoppers were transferred to rice plants 24 hours after the gene's dsRNA was detected, and their acquired body weight over 24 hours was recorded. Results are as follows: Figure 4 As shown in B, ds Nllipase1 Group weight ratio control group and ds GFP The group showed a significant decrease. This indicates that the silent brown planthopper... Nllipase1 Afterwards, the honeydew production of brown planthoppers decreased significantly; and ds GFP There was no significant difference between the control group and the control group. Based on the above results, Nllipase1 It is closely related to the amount of food consumed by brown planthoppers.

[0077] Example 7: Silence Nllipase1 Impact on the mortality rate of brown planthoppers

[0078] Injection Nllipase1 Brown planthoppers, after receiving dsRNA from the gene, fed on rice. Mortality rates were recorded every other day for 15 days. The results are as follows: Figure 5 As shown. ds Nllipase1 The mortality rate in the control group was significantly higher than that in the control group and the ds group. GFP The group, and by the 5th day, it had exceeded 50%.

[0079] SEQ ID NO.1 Atgtcgtctgattgcttcctcgttgataccgatggattaggttttgtactagctcgagacaactgggatgtgtggcttgggaatattagaggaaattattacagtagagcacataacagaatatcacctaacagctctgagttttggaactacagttggcacgacatgggccagaaagatataaccgccatgattgattacatactcaataaaactggaaaaccgcaactgaactacatcggccactcaatgggaaccacaatgttctatgtgatggcatccactatgcccagctacaacaagaagatcaacaccatgataagtttggctccaatcggcagaatgactaaatggaactttgtactcaataactcgacttacatttaccagtacacacatgattacactatgaaaaatattcgccaaaaaaaggttgaagttctgcctagaggagagtttaaaggaatgtccaagcatcaaattctaatggaagatttggacacctacttcaggactacatttgagattgtaacttggtttgttaattcagtgactggtgaagacatatttccagacggtacttctaccagaaacttactacattatcttcaaaatattggatcaggaacatttcaaaggtttgattacggttcacgtgagaggaacatgaaggcttggggacatgaagctccaacaccatacaatttaaacaacattaaagcgcctgtagccgtgttttacgcagtagcagatgaattcgcgtccgaaaaagacgcggagtatatgatttcaaaattgccaaacgttgtgatgaaaatgaaaatttcagctatgcgatttggacacggtgatttctgcttcactactgccaaagagaacctcagcatgcgtcgaaagatgcacgcctccattttgaatctattgaaacagcgagatgaacaaagtgttgccaaaaaatcttgtcagaacaagaaagattgtgatccaaaatcgtaa

[0080] SEQ ID N0.2 MSSDCFLVDTDGLGFVLARDNWDVWLGNIRGNYYSRAHNRISPNSSEFWNYSWHDMGQKDITAMIDYILNKTGKPQLNYIGHSMGTTMFYVMASTMPSYNKKINTMISLAPIGRMTKWNFVLNNSTYIYQYTHDYTMKNIRQKKVEVLPRGEFKGMSKHQILMEDLDTYFRTTFEIVTWFVNSVTGEDIFPDGTSTRNLLHYLQNIGSGTFQRFDYGSRERNMKAWGHEAPTPYNLNNIKAPVAVFYAVADEFASEKDAEYMISKLPNVVMKMKISAMRFGHGDFCFTTAKENLSMRRKMHASILNLLKQRDEQSVAKKSCQNKKDCDPKS

Claims

1. A type of brown planthopper Nllipase1 Genes, characterized by, The Nllipase1 The nucleotide sequence of the gene is shown in SEQ ID NO.

1.

2. A type of brown planthopper Nllipase1 Gene-encoded proteins are characterized by, The amino acid sequence is shown in SEQ ID NO.

2.

3. A type of brown planthopper Nllipase1 The dsRNA of a gene is characterized by, The sequence shown in SEQ ID NO.1 can achieve inhibition. Nllipase1 A double-stranded RNA composed of nucleotides that affect gene expression and their inverse complementary nucleotide sequences.

4. The dsRNA according to claim 3, characterized in that, The brown planthopper as described in claim 1 Nllipase1 Using genes as templates, ds Nllipase1 -F, ds Nllipase1 -R is used as a primer for PCR amplification to obtain the PCR amplification product, and then dsRNA is synthesized. ds Nllipase1 -F: 5'- GGATCCTAATACgACTCACTATAGG ccgatggattaggttttgtact-3' ds Nllipase1 -R: 5'- GGATCCTAATACGACTCACTATAGG acgattttggatcacaatctttcttg-3'。 5. The use of the dsRNA according to claim 3 or 4 in the preparation of a product, wherein the product is used in any one of the following applications (1)-(3): 1) Controlling brown planthoppers or preparing products for controlling brown planthoppers; 2) Reduce the survival rate of brown planthoppers or prepare products that reduce the survival rate of brown planthoppers; 3) Suppressing brown planthopper genes Nllipase1 Gene expression or preparation to inhibit brown planthoppers Nllipase1 Products that express genes.

6. A method for controlling brown planthoppers, characterized in that, The dsRNA described in claim 3 or 4 is introduced into brown planthoppers to control brown planthoppers, reduce their survival rate, or inhibit their growth. Nllipase1 Gene expression.

7. The method according to claim 6, characterized in that, The method of introduction was microinjection, in which dsRNA was injected into the brown planthopper, which then fed on it. The results showed that both the amount of food consumed and the survival rate of the rice planthopper decreased.

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