MiR-10490-5p for preventing and treating spodoptera frugiperda and application of miR-10490-5p
By targeting and inhibiting the JHEH gene of fall armyworm with miR-10490-5p and agomiR-10490-5p, the problem of drug resistance caused by chemical control was solved, and safe and efficient control of fall armyworm was achieved.
Patent Information
- Application Number
- CN202511529388.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-10-24
- Publication Date
- 2026-03-03
AI Technical Summary
Current technologies for controlling fall armyworm mainly rely on chemical control, which leads to pesticide resistance and is harmful to the ecology and food safety. There is a lack of effective green control methods.
Using miR-10490-5p and its analogue agomiR-10490-5p, the expression of the juvenile hormone epoxide hydrolase (JHEH) gene in fall armyworm was targeted and inhibited by injection or feeding, affecting insect growth and development, feeding amount, and increasing larval mortality.
It effectively inhibits the growth, development, and feeding of the fall armyworm, increases larval mortality, and provides a safe and efficient control strategy.
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Figure CN121592649A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of agricultural biotechnology, specifically, it relates to a method for controlling the fall armyworm. Spodoptera frugiperda miR-10490-5p and its applications. Background Technology
[0002] MicroRNAs (miRNAs) are a class of single-stranded non-coding small RNAs, 18-24 nucleotides in length, encoded by endogenous genes and widely found in eukaryotic cells and viruses. These miRNAs regulate gene expression in plant and animal cells by binding to target mRNAs through base pairing, leading to silencing. In animals, they regulate the expression of more than half of the genes encoding proteins. A single miRNA can target hundreds of mRNAs, and multiple miRNAs can regulate the same mRNA. Besides regulating the expression of protein-coding genes, the expression of miRNAs themselves can also be regulated by other miRNAs. miRNAs bind to the 3' untranslated region (3'UTR) or open reading frame (ORF) of the target mRNA, inhibiting translation or promoting mRNA degradation, thereby regulating gene expression. Furthermore, miRNAs exhibit high conservation in both gene location and sequence. For example, approximately 12% of miRNAs are highly conserved in model organisms such as nematodes and fruit flies, with base differences of only 1-2 nucleotides, demonstrating their important evolutionary function.
[0003] fall armyworm Spodoptera frugiperda The fall armyworm (PMO) is a lepidopteran noctuid moth originating from tropical and subtropical regions of the Americas. It is characterized by its wide host range, high reproductive capacity, rapid migration and spread, and severe damage. The fall armyworm primarily feeds on gramineous crops such as corn and rice, as well as cruciferous and solanaceous vegetables, causing serious harm to global agricultural production. In 2019, the fall armyworm invaded my country and rapidly established itself and spread in the southwest region. By October of the same year, it had spread to nearly 30 provinces and municipalities across the country. Currently, the control of the fall armyworm mainly relies on chemical control. However, the extensive use of chemical pesticides has led to pesticide resistance in the fall armyworm and has also had adverse effects on ecological and food safety. Therefore, developing effective green control technologies for the fall armyworm has become an urgent problem to be solved. RNA interference technology, which specifically silences key genes that play an important role, provides an efficient and safe green control approach for the fall armyworm. However, there is still very little research and technology on identifying effective target genes and using RNAi to control the fall armyworm.
[0004] Juvenile hormone epoxide hydrolase (JHEH) degrades juvenile hormone into juvenile hormone diol (JHd), thereby regulating juvenile hormone levels in insects. During specific developmental stages, the activity of JHEH significantly increases, breaking down juvenile hormone and leading to a decrease in hormone levels. This decrease in juvenile hormone concentration allows insects to transition from larval to pupal or adult stages. Inhibition of JHEH expression leads to increased juvenile hormone concentrations, hindering insect growth and development. For example, silencing JHEH expression in the cotton bollworm (…) using RNAi… Helicoverpa armigera Diamondback moth ( Plutella xylostella ) or brown planthopper ( Nilaparvata lugens After JHEH treatment, the mortality rate of larvae in the treatment group increased significantly by more than 50% compared to the control group. Summary of the Invention
[0005] The purpose of this invention is to provide a miR-10490-5p for the control of fall armyworm and its application, and to further synthesize its analogue agomiR-10490-5p. Injection of agomiR-10490-5p revealed its inhibitory effect on the expression of the juvenile hormone cyclohydrolase (JHEH) gene (NCBI ID XM_035594190.2), a key gene for the growth and development of fall armyworm. Furthermore, both injection and feeding methods demonstrated that agomiR-10490-5p can inhibit the growth and development of fall armyworm, reduce its food intake, and increase the mortality rate of fall armyworm larvae. This miRNA provides a new target and strategy for the control of fall armyworm and has good application potential.
[0006] The specific plan is as follows: A miR-10490-5p inhibitor for controlling night starvation in grasslands has the following nucleotide sequence: 5'-UUCCAACGAAUGCAAGACCG-3'.
[0007] A miR-10490-5p analogue for controlling night starvation in grasslands, namely agomiR-10490-5p, has a double-stranded structure, and its nucleotide sequence is shown below: .
[0008] The application of miR-10490-5p or its analogues described in this invention in the preparation of drugs for controlling fall armyworm.
[0009] A method for preventing and controlling night starvation in grasslands, using miR-10490-5p or similar substances described in this invention, by means of injection.
[0010] Optionally, the dosage of the active ingredient in the injection method is 600 ng / insect.
[0011] A method for preventing night starvation in grasslands, using miR-10490-5p or similar products as described in this invention, employs a feeding method to prevent night starvation in grasslands.
[0012] Optionally, the feeding time for the effective ingredient in the feeding method is 12 hours, and the dosage is 1 to 1.5 ug / insect.
[0013] The method for synthesizing miR-10490-5p analogs for controlling fall starvation in grasslands as described in this invention includes: modifying the 3' end of the antisense strand with cholesterol, modifying the 5' end with two thiosclerotic backbones, modifying the 3' end with four thiosclerotic backbones, and modifying the entire chain with methoxy groups according to the nucleotide sequence of miR-10490-5p, thereby completing the chemical synthesis.
[0014] A kit for controlling fall armyworm, the kit comprising miR-10490-5p as described in this invention or an analogue as described in this invention.
[0015] A drug for preventing grassland night starvation, wherein the drug contains miR-10490-5p as described in this invention or an analogue as described in this invention.
[0016] The beneficial effects obtained by this invention are as follows: This invention provides a miRNA for controlling fall armyworm and its application, specifically miR-10490-5p. A double-stranded miRNA analog, agomiR-10490-5p, was also chemically synthesized. The targeting of fall armyworm miR-10490-5p to the key growth and development gene JHEH was detected using a dual-luciferase assay (in vitro verification) and quantitative real-time PCR (in vivo verification), demonstrating that this miRNA effectively inhibits the expression of the target gene JHEH. Using the miRNA and its analog, fall armyworm was controlled via injection or feeding, demonstrating that both the miRNA and its analog effectively control the fall armyworm. Attached Figure Description
[0017] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the following detailed description to explain the invention, but do not constitute a limitation thereof. In the drawings: Figure 1A schematic diagram illustrating how miR-10490-5p regulates insect growth and development by targeting the JHEH gene and predicting the binding site of miR-10490-5p to the target gene. Figure 2 The diagram illustrates the in vitro validation analysis of the interaction between miR-10490-5p and its target gene JHEH; luciferase activity of NC and JHEH interactions is used as 1, and *** indicates significant differences ( ). P <0.001), ** indicates a significant difference ( P <0.01), * indicates a significant difference ( P <0.05), ns indicates that there is no significant difference ( P >0.05). The luciferase activity of the mutant JHEH interacting with miR-10490-5p was used as a control; Figure 3 This is a diagram illustrating the in vivo validation analysis of the interaction between miR-10490-5p and the target gene JHEH; the expression levels of miRNA / target gene in *Pseudomonas aeruginosa* injected with / fed with NC are used as 1, and *** indicates significant differences. P <0.001), ** indicates a significant difference ( P <0.01), * indicates a significant difference ( P <0.05), ns indicates that there is no significant difference ( P >0.05); Figure 4 The diagram illustrates the effect of injection and feeding of agomiR-10490-5p on the body length of the fall armyworm; *** indicates a significant difference. P <0.001), ** indicates a significant difference ( P <0.01), * indicates a significant difference ( P <0.05), ns indicates that there is no significant difference ( P >0.05); Figure 5 The graph illustrates the effect of injection and feeding of agomiR-10490-5p on the feed intake of fall armyworm; *** indicates significant difference. P <0.001), ** indicates a significant difference ( P <0.01), * indicates a significant difference ( P <0.05), ns indicates that there is no significant difference ( P >0.05); Figure 6 A graphical representation of the effect of injection and feeding of agomiR-10490-5p on the mortality rate of fall armyworm; *** indicates a significant difference ( P<0.001), ** indicates a significant difference ( P <0.01), * indicates a significant difference ( P <0.05), ns indicates that there is no significant difference ( P >0.05). Detailed Implementation
[0018] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. The present invention will be further described below with reference to implementation examples. It should be noted that these embodiments are only used to explain the present invention and are not intended to limit the scope of the present invention.
[0019] The miRNA provided by this invention for controlling fall armyworm, namely miR-10490-5p, can target the juvenile hormone epoxide hydrolase gene (JHEH), a key gene for the growth and development of fall armyworm. The nucleotide sequence of miR-10490-5p is shown in SEQ ID NO: 1. An analogue of miR-10490-5p, agomiR-10490-5p, was synthesized in vitro via chemical synthesis. The double-stranded sequences are shown in SEQ ID NO: 1 and SEQ ID NO: 2. Injection and feeding methods revealed the inhibitory effects of the analogue agomiR-10490-5p on the growth, development, and feed intake of fall armyworm, as well as its lethal effect. This miRNA provides a new target and strategy for the control of fall armyworm, and has good application potential. This invention clarifies for the first time the negative regulatory mechanism of miR-10490-5p of fall armyworm on the target gene JHEH, and clarifies the inhibitory effect of the miR-10490-5p analog agomiR-10490-5p on the growth, development and food intake of fall armyworm, as well as its lethal effect on fall armyworm larvae.
[0020] The miR-10490-5p of the present invention for controlling fall armyworm has the following nucleotide sequence: 5'-UUCCAACGAAUGCAAGACCG-3' (SEQ ID NO: 1).
[0021] This invention provides a miRNA analog for miRNA overexpression, namely agomiR-10490-5p, which is a chemically synthesized double-stranded structure with the following nucleotide sequences (SEQ ID NO: 1 and SEQ ID NO: 2): SEQ ID NO: 1: 5'-UUCCAACGAAUGCAAGACCG-3'; SEQ ID NO: 2: 5'-UUAAGGUUGCUUACGUUCUG-3'.
[0022] .
[0023] This invention provides the nucleotide sequence of the juvenile hormone epoxide hydrolase JHEH gene, a target gene of miR-10490-5p (GenBank accession number: XM_035594190.2).
[0024] This invention provides a method for verifying the interaction between miRNA and target gene, specifically a method for verifying the interaction between miR-10490-5p and the target gene JHEH, comprising the following steps: S1 In vitro validation: In vitro validation was achieved using a dual-luciferase assay. First, the target gene sequence was synthesized and constructed into the pmirGLO vector, with a mutated target gene (the fragment TCAGCGTATCGTTGGA at the miR-10490-5p binding site of the target gene JHEH mutated to CTAGGACGCTACCAAG) used as a control. miR-10490-5p analogs (small double-stranded miRNA fragments designed and synthesized targeting the mature form of the miRNA, with the same function as the mature miRNA) and an NC control were synthesized. The plasmid and miRNA were co-transfected into Sf9 cells and cultured. Following the steps described in the dual-luciferase reporter gene assay kit, luciferase activity was detected by measuring the fluorescence intensity at 350-700 nm to analyze the regulatory effect of the miRNA on the target gene.
[0025] S2 In vivo validation: In vivo validation was achieved by introducing agomiR-10490-5p into the bodies of fall armyworm larvae. The injection method involved diluting agomiR-10490-5p to a concentration of 1 μg / μL and injecting 0.6 μL of agomiR-10490-5p into the abdomen of early third instar fall armyworms using a microsyringe. The feeding method involved adding 50 μg of agomiR-10490-5p analog to every 1 g of artificial feed. After 12 hours of feeding, the fall armyworms were removed and subsequently fed clean artificial feed. Sufficient samples were collected 48 hours after treatment, frozen in liquid nitrogen, and then stored at -80°C for subsequent experiments. miRNA and total RNA were extracted from the injected fall armyworms according to the kit, and reverse transcribed for real-time quantitative PCR. The expression levels of miR-10490-5p and the target gene JHEH were detected.
[0026] This invention provides a method for preparing miRNA or miRNA analogs through chemical synthesis. The method for preparing agomiR-10490-5p involves modifying the 3' end of the antisense strand with cholesterol, two thiocarbamates at the 5' end, four thiocarbamates at the 3' end, and a full-chain methoxy group based on the nucleotide sequence of miR-10490-5p, followed by modification of the entire antisense strand with cholesterol, two thiocarbamates at the 5' end, four thiocarbamates at the 3' end, and a full-chain methoxy group, thus completing the chemical synthesis. The synthesized agomiR-10490-5p exhibits stable structure, long expression time, and requires no transfection reagents.
[0027] The present invention provides nucleic acid molecular sequences encoding miRNAs or similar molecules.
[0028] This invention provides the application of miRNA or analogues in the preparation of pesticides for controlling fall armyworm.
[0029] The present invention provides a formulation or kit for controlling fall armyworm, wherein the formulation or kit contains any one of the miRNAs or analogs of the present invention.
[0030] This invention provides a method for controlling fall armyworm, which utilizes miRNA and analogues and employs injection or feeding methods to control fall armyworm.
[0031] The concentration of the analogue in the injection method is 600 ng / head.
[0032] The dosage of the analog in the feeding method is 50 μg agomiR-10490-5p analog per 1 g of artificial feed.
[0033] This invention provides a method for quantitative fluorescence detection of miRNA, specifically a method for quantitative fluorescence detection of miR-10490-5p expression levels in fall armyworm larvae, comprising the following steps: S1: Extract miRNA from fall armyworms that were injected / fed agomiR-10490-5p and the control group 48 h after treatment; S2: The expression level of miR-10490-5p in fall armyworm at each time point was determined by real-time quantitative PCR.
[0034] The primers used to detect miR-10490-5p expression were: forward primer 5'-GCTTCCAACGAATGCAAGACCG-3', reverse primer 5'-GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGAC-3'; for the internal reference gene (U6snRNA), the forward primer was 5'-GCTTCGGCAGCACATATACTAAAAT-3', and the reverse primer was 5'-CGCTTCACGAATTTGCGTGTCAT-3'.
[0035] This invention provides a method for quantitative fluorescence detection of target genes of miRNA, specifically a method for quantitative fluorescence detection of the expression level of the miR-10490-5p target gene JHEH in fall armyworm larvae, comprising the following steps: S1: RNA was extracted from fall armyworms 48 h after injection / feeding agomiR-10490-5p and control treatment, and cDNA was obtained by reverse transcription. S2: The expression level of the JHEH gene in the fall armyworm was determined by real-time quantitative PCR.
[0036] The primers used in S1 are as follows: For the detection of JHEH expression level, the primers are: forward primer 5'-TCTGAGGGAACGAATAAACGAGGAC-3', reverse primer 5'-GCCATGTGTGAAGCGACTTGTG-3'; for the internal reference gene (β-actin), the forward primer is 5'-ATCCTCCGTCTGGACTTGG-3', and the reverse primer is 5'-CGCACGATTTCCCTCTCA-3'.
[0037] The amplification program used was as follows: 94℃ pre-denaturation for 30s; 94℃, 5s, 60℃, 30s, for 45 cycles; 95℃, 15s, 60℃, 1min (increasing by 0.3℃ each time until reaching 95℃); 95℃, 15s; the reaction volume was 20μL: forward and reverse primers 0.4μL, enzyme buffer 10μL, cDNA 1μL and ddH2O 8.2μL.
[0038] Experimental materials and reagents 1. Cells and vectors: Sf9 cells were purchased from Invitrogen. pmirGLO vector was purchased from Wuhan Jinkairui Biotechnology Co., Ltd.
[0039] 2. Enzymes and kits: The enzymes required for PCR were purchased from Beijing TransGen Biotech Co., Ltd., and the miRNAs, miRNA analogs, and primers used were synthesized by Sangon Biotech (Shanghai) Co., Ltd.
[0040] Note: Molecular biology experimental methods not specifically described in the following examples were performed in accordance with the specific methods listed in J. Sambrook's "Molecular Cloning: A Laboratory Manual," 3rd edition, or according to the kit and product instructions.
[0041] Example 1: In vitro verification of the interaction between miR-10490-5p and the target gene JHEH In vitro validation was achieved using a dual-luciferase assay. The target gene sequence was synthesized and constructed into the pmirGLO vector (containing Firefly luciferase and Renilla luciferase), with a mutated target gene (the fragment at the miR-10490-5p binding site of the target gene JHEH mutated from TCAGCGTATCGTTGGA to CTAGGACGCTACCAAG) used as a control. miR-10490-5p mimics and an NC control were synthesized; the plasmid and miRNA were transfected into Sf9 cells and cultured. Luciferase activity was detected by measuring the fluorescence intensity at 350-700 nm, following the steps described in the dual-luciferase reporter gene assay kit. Figure 2 The results showed that overexpression of miR-10490-5p could significantly reduce the expression level of JHEH, meaning that miR-10490-5p can target JHEH and negatively regulate the expression of JHEH.
[0042] Example 2: In vivo verification of the interaction between miR-10490-5p and the target gene JHEH In vivo validation was achieved by introducing agomiR-10490-5p into the bodies of fall armyworm larvae. Injection validation: agomiR-10490-5p was diluted to a concentration of 1 μg / μL, and 0.6 μL of agomiR-10490-5p was injected into the abdomen of early third instar fall armyworms using a microsyringe. Feeding validation: 50 μg of agomiR-10490-5p analog was added to every 1 g of artificial feed. After 12 h of feeding, the fall armyworms were removed and fed clean artificial feed. Forty-eight h after treatment, sufficient samples were collected, frozen in liquid nitrogen, and stored at -80°C for subsequent experiments. miRNA and total RNA were extracted from the injected fall armyworms according to the kit, and reverse transcribed for real-time quantitative PCR. The expression levels of miR-10490-5p and the target gene JHEH were detected. Real-time quantitative PCR was used to record experimental data, and the relative expression levels of the target gene were calculated using the 2-ΔΔCT method. The relative gene expression levels were analyzed using an independent samples t-test. Statistical analysis was performed using Excel 2013 and SPSS 16. Figure 3 The results showed that after 48 h of injection and feeding treatment, the expression levels of miR-10490-5p were 5.52-fold and 4.03-fold higher than those of the control treatment, respectively, while the expression levels of the target gene JHEH were 0.0054-fold and 0.13-fold higher than those of the control treatment, respectively. This indicates that the miR-10490-5p analogue agomiR-10490-5p can significantly reduce the expression level of JHEH, meaning that miR-10490-5p can target JHEH and negatively regulate its expression.
[0043] Example 3: Determination of body length of fall armyworm after injection and feeding with agomiR-10490-5p AgomiR-10490-5p was introduced into the larvae of the fall armyworm using both injection and feeding methods. For the injection method, agomiR-10490-5p was diluted to a concentration of 1 μg / μL, and 0.6 μL of agomiR-10490-5p was injected into the abdomen of early third instar fall armyworms using a microsyringe. For the feeding method, 50 μg of agomiR-10490-5p analogue was added to every 1 g of artificial feed. After 12 hours of feeding, the fall armyworms were removed and subsequently fed clean artificial feed. The body length of surviving fall armyworms was measured at 24 h, 48 h, 72 h, and 96 h post-treatment. Each treatment had three replicates, with 20 fall armyworm larvae per replicate. The mean values of each treatment group were calculated, and independent samples t-tests were used for difference analysis. Statistical analysis software included Excel 2013 and SPSS 16. Figure 4 The results showed that at all four time points after injection and feeding treatments, the body length of the fall armyworm was significantly different from that of the control group. P <0.05), and 96 h after treatment, the body length of fall armyworm in the treatment group was 0.66 times and 0.81 times that of the control group, respectively. This indicates that injection or feeding of the miR-10490-5p analogue agomiR-10490-5p can significantly inhibit the growth of fall armyworm larvae.
[0044] Example 4: Determination of feed intake of fall armyworm after injection and feeding agomiR-10490-5p AgomiR-10490-5p was introduced into the larvae of the fall armyworm using both injection and feeding methods. For the injection method, agomiR-10490-5p was diluted to a concentration of 1 μg / μL, and 0.6 μL of agomiR-10490-5p was injected into the abdomen of early third instar fall armyworms using a microsyringe. For the feeding method, 50 μg of agomiR-10490-5p analogue was added to every 1 g of artificial feed. After 12 hours of feeding, the fall armyworms were removed and subsequently fed clean artificial feed. The daily feed intake of surviving fall armyworms was recorded at 24 h, 48 h, 72 h, and 96 h after treatment. Each treatment had three replicates, with 20 fall armyworm larvae per replicate. The mean values of each treatment group were calculated, and independent samples t-tests were used for difference analysis. Statistical analysis software included Excel 2013 and SPSS 16. Figure 5 The results showed that at all four time points after injection and feeding treatments, the daily food intake of fall armyworm was significantly different from that of the control group.P <0.05), and 96 h after treatment, the daily food intake of fall armyworm in the treatment group was 0.67 times and 0.73 times that of the control group, respectively. This indicates that injection or feeding of the miR-10490-5p analogue agomiR-10490-5p can significantly inhibit the food intake of fall armyworm larvae.
[0045] Example 5: Determination of mortality rate of fall armyworm after injection and feeding of agomiR-10490-5p AgomiR-10490-5p was introduced into the larvae of the fall armyworm via injection and feeding. For the injection method, agomiR-10490-5p was diluted to a concentration of 1 μg / μL, and 0.6 μL of agomiR-10490-5p was injected into the abdomen of early third instar fall armyworms using a microsyringe. For the feeding method, 50 μg of agomiR-10490-5p analogue was added to every 1 g of artificial feed. After 12 hours of feeding, the fall armyworms were removed and subsequently fed clean artificial feed. Mortality rates were recorded at 24 h, 48 h, 72 h, and 96 h post-treatment. Each treatment had three replicates, with 20 fall armyworms per replicate. The mean values for each treatment group were calculated, and the mortality rates were transformed using the arcsine square root and analyzed using an independent samples t-test. Statistical analysis was performed using Excel 2013 and SPSS 16. Figure 6 The results showed that at all four time points after injection and feeding treatments, the mortality rate of fall armyworm was significantly different from that of the control group. P <0.05), and 96 h after injection and feeding, the mortality rates of fall armyworm in the treatment groups were 83.33% and 63.33%, respectively, which were 66.66% and 55.00% higher than those in the control group. This indicates that injection or feeding of the miR-10490-5p analogue agomiR-10490-5p can significantly increase the mortality rate of fall armyworm larvae.
[0046] The above description is merely a preferred embodiment of the present invention. It should be noted that, due to the limitations of written expression, there are objectively infinite specific structures. For those skilled in the art, several improvements, modifications, or variations can be made without departing from the principle of the present invention, and the above technical features can be combined in an appropriate manner. These improvements, modifications, variations, or combinations, or the direct application of the inventive concept and technical solution to other situations without modification, should all be considered within the scope of protection of the present invention.
Claims
1. A miR-10490-5p for controlling night starvation in grasslands, characterized in that, Its nucleotide sequence is shown below: 5'-UUCCAACGAAUGCAAGACCG-3'.
2. A miR-10490-5p analogue for controlling fall starvation in grasslands, characterized in that, Its name is agomiR-10490-5p, which is a double-stranded structure, and its nucleotide sequence is shown below: 。 3. The use of miR-10490-5p as described in claim 1 or the analogue as described in claim 2 in the preparation of a drug for controlling fall armyworm.
4. A method for preventing grassland from starving overnight, characterized in that, Using miR-10490-5p as described in claim 1 or an analogue as described in claim 2, the grassland night starvation can be controlled by injection.
5. The method for preventing grassland starvation according to claim 4, characterized in that, The dosage of the active ingredient in the injection method is 600 ng / insect.
6. A method for preventing grassland from starving overnight, characterized in that, Using miR-10490-5p as described in claim 1 or similar substances as described in claim 2, a feeding method is employed to prevent night starvation in grasslands.
7. The method for preventing grassland starvation according to claim 6, characterized in that, The feeding time for the effective ingredient in the feeding method is 12 hours, and the dosage is 1-1.5 ug / insect.
8. The method for synthesizing the miR-10490-5p analogue for controlling fall starvation in grasslands as described in claim 2, characterized in that, include: Based on the nucleotide sequence of miR-10490-5p, the 3' end of the antisense strand was modified with cholesterol, the 5' end with two thiosclerotic backbones were modified, the 3' end with four thiosclerotic backbones were modified, and the entire chain was modified with methoxy groups to complete the chemical synthesis.
9. A kit for controlling fall armyworm, characterized in that, The kit comprises miR-10490-5p as described in claim 1 or an analogue as described in claim 2.
10. A drug for preventing grassland from starving at night, characterized in that, The drug contains miR-10490-5p as described in claim 1 or an analogue as described in claim 2.