Application of biological materials regulating P120 catenin gene expression in locust control

Abstract

The invention belongs to the field of biotechnology, relates to locust control, and particularly refers to the application of biological materials for regulating the expression of P120 catenin genes in locust control. The present invention can specifically silence the P120 catenin gene of locusts by injecting the dsRNA of the synthesized P120 catenin gene into the locust body cavity, thereby inhibiting the oviposition of locusts and causing a large number of deaths, with a mortality rate of 98.4%, thereby effectively inhibiting the growth of locust populations and achieving good results in preventing and controlling locusts. Due to the high efficiency, specificity and safety of the present invention, the present invention has broad application prospects in locust pest control, and has good economic benefits for protecting my country's agricultural production and ecological safety.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, relates to locust control, and particularly refers to the application of biological materials for regulating the expression of P120 catenin genes in locust control. Background Art

[0002] Locust (Locusta migratoria), Insecta, Orthoptera, Acrididae, is an important crop pest, commonly known as grasshopper. There is only one species, but due to different geographical distribution, there are 9 known subspecies in the world. There are 3 subspecies distributed in China, namely the East Asian migratory locust, the Asian migratory locust and the Tibetan migratory locust, represented by the East Asian migratory locust, which is the most widely distributed and the most harmful. Migratory locusts have a mixed diet, and adults and locust nymphs mainly harm wheat, corn, rice, millet, sorghum, millet, and wild grass plants such as reeds, barnyard grass, reeds and Cyperus trifoliate. The East Asian migratory locust is an important agricultural pest in Southeast Asia and a major historical pest in China. From the Spring and Autumn Period of the Zhou Dynasty to around 1950, there was an average of a large regional outbreak every 2 to 3 years, and a larger-scale rampant outbreak every 5 to 7 years. The main affected areas are mostly agricultural areas in the eastern Huanghuai Plain, alternating with drought and flood disasters in the region. There are many historical records of the tragic scenes of locust plagues. Before the end of the 1940s, there were three major locust plagues, namely in 1929, 1933 and 1938, which spread to 10 provinces (cities) and 265 counties. By the early 1940s, locust plagues occurred almost every year. Under normal circumstances, adults can mate on average about 7 days after emergence. Female insects can mate 20 to 25 times in their lifetime. They can lay eggs under the soil 4 to 7 days after mating, with a depth of about 4 to 6 cm. When the eggs are laid, mucus is secreted to glue the eggs to form egg masses. Each female insect can generally lay 4 to 5 egg capsules, up to 12 pieces, each of which generally contains 50 to 80 eggs, and the total number of eggs laid in a lifetime is an average of 300 to 400. When laying eggs, they have a certain selectivity for terrain, orientation, vegetation and the physical and chemical properties of the soil, and most of them are laid in relatively solid soil. The most suitable soil moisture content for egg laying is 10-20%. When the soil moisture content is lower than 5% or higher than 25%, the number of eggs laid is significantly reduced. The suitable area for egg laying is the area with a chloride ion content of 0.2-1.2%. The most eggs are laid in areas with a salt content of less than 0.5%. The minimum salt content of the egg laying area is 0.3%. When the salt content of the surface soil is higher than 1.2%, there is no distribution of eggs. Parthenogenesis is also possible. The hatched young nymphs are female, grow and develop normally, and can still mate and lay eggs after emergence. This is an important reproductive characteristic for locusts to self-regulate when the population is extremely low.

[0003] The use of dsRNA to control pests has the following advantages: dsRNA only degrades homologous mRNA, reducing the toxic effects on non-target organisms, especially for higher animals and humans. A small amount of dsRNA can effectively inhibit the expression of target genes, and dsRNA entering the body of pests can cause systemic silencing of target genes and even in offspring. RNA is easily degraded in nature, leaving no residue, and is relatively safe and non-polluting to the ecological environment and crop products. At present, the basic technology for pest control based on RNA interference is to screen target sequences to obtain dsRNA with high lethality. The patent with publication number CN116769810A discloses the TRE gene of the Asian locust, which causes the death of the Asian locust by infecting its chitin metabolism; the application with publication number CN112662690A discloses the dsRNA of the migratory locust Rab5 gene for the prevention and control of migratory locusts, and the migratory locust Rab5 gene is a key regulator of cell transport; the patent with publication number CN112662689A discloses a migratory locust Rab11A gene and its dsRNA for the prevention and control of migratory locusts, and the migratory locust Rab11A gene is a migratory locust small GTPase gene. The above methods all interfere with the target gene to kill the target migratory locust. This research group is committed to exploring undeveloped functional genes in the migratory locust genome in order to find more efficient targets for the prevention and control of migratory locusts. Summary of the invention

[0004] The present invention proposes an application of a biological material for regulating the expression of the P120 catenin gene in the prevention and control of migratory locusts. The dsRNA synthesized from the migratory locust P120 catenin gene is injected into the body of female migratory locusts, which can prevent the female migratory locusts from laying eggs and cause death, thereby preventing the migratory locusts from breaking out and causing disasters.

[0005] The technical solution of the present invention is achieved in this way:

[0006] Application of biological materials that regulate P120 catenin gene expression in locust control.

[0007] The ORF sequence of the P120 catenin gene is shown in SEQ ID No.1. The ORF sequence of the P120 catenin gene is 963 bp in length and encodes a 320 amino acid sequence as shown in SEQ ID No.14.

[0008] The above-mentioned biological material for regulating the expression of P120 catenin gene is a biological material for inhibiting or reducing the expression of P120 catenin gene.

[0009] The above-mentioned regulatory biological material is an RNA molecule that inhibits or reduces the expression of the P120 catenin gene.

[0010] The RNA molecule is a double-stranded RNA molecule, comprising a sense strand nucleotide as shown in SEQ ID No.9 and an antisense strand nucleotide that is reverse complementary to the sequence shown in SEQ ID No.9.

[0011] The above-mentioned biological material is a DNA molecule that produces the above-mentioned RNA molecule.

[0012] The sequence of the above DNA molecule is shown as SEQ ID No.6.

[0013] The method for synthesizing double-stranded RNA molecules using the above DNA molecules is:

[0014] (1) using the cDNA reverse transcribed from the total RNA of the fat body of the locust as a template, PCR amplification was performed using the upstream and downstream primers shown in SEQ ID No. 4 and SEQ ID No. 5 to obtain the PCR amplification product shown in SEQ ID No. 6;

[0015] (2) The PCR amplification product shown in SEQ ID No. 6 was connected into the pGEM-T vector to obtain the recombinant plasmid pGEM-T-P120 catenin;

[0016] (3) using the recombinant plasmid pGEM-T-P120 catenin as a template, PCR amplification was performed using a primer pair with a T7 promoter sequence as shown in SEQ ID No. 7 and SEQ ID No. 8 to obtain a DNA template for synthesizing dsRNA (i.e., DNA with a T7 promoter sequence);

[0017] (4) Using the DNA template used to synthesize dsRNA in step (3), use T7RiboMAX TM dsRNA was synthesized by in vitro transcription using the Express RNAiSystem (Promega) kit.

[0018] A method for preventing female migratory locusts from laying eggs and / or causing them to die, the method comprising treating female adult migratory locusts with the double-stranded RNA molecules described above.

[0019] The above-mentioned female adult locusts are female adult locusts within 12 hours of emergence.

[0020] The dosage of the double-stranded RNA molecule was 15 μg per mouse.

[0021] The present invention has the following beneficial effects:

[0022] 1. The present invention can effectively inhibit the gene expression of P120 catenin by injecting the synthetic dsRNA based on the locust P120 catenin gene into the female adult locust, produce the effects of inhibiting egg laying and causing death, and the dosage of dsRNA is small (15 μg), the lethality rate can reach 98.4%, and the effect is remarkable. In addition, the use of dsRNA to control locusts has strong species specificity and will not cause gene interference to other species; at the same time, dsRNA is easily degraded in the natural environment and is friendly to the ecological environment. Therefore, in view of the high efficiency, specificity and safety of the present invention, it has broad application prospects in the control of locust pests and has good economic benefits for protecting my country's agricultural production and ecological security. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0024] Figure 1 This is the interference efficiency of P120 catenin gene in fat body and ovarian tissue after double-stranded RNA injection; ***P<0.001 in the figure.

[0025] Figure 2 This is a statistical chart of the survival rate of locust adults after interfering with the expression of P120 catenin. DETAILED DESCRIPTION

[0026] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] Example 1: Sequence of Locust P120 catenin gene and its dsRNA

[0028] 1. Obtaining the P120 catenin gene sequence of Locust migratory locust

[0029] 1.1 Anatomy of Locust Tissue

[0030] Select healthy female adult locusts, first cut off their heads with anatomical scissors that have been sterilized with alcohol, then cut open the abdomen from the cut site, use anatomical forceps that have been sterilized with alcohol to remove the fat bodies and ovaries in the abdominal cavity and put them into 2 mL centrifuge tubes respectively, quickly place the centrifuge tubes in liquid nitrogen for instant freezing, and store them in a -80°C refrigerator for later use.

[0031] 1.2 Total RNA extraction

[0032] (1) Place the tissue obtained above on ice, add 300 μL TRIzol (Tiangen), and grind it thoroughly on ice with an electric grinding rod; let the tissue suspension stand at room temperature for 30 minutes to ensure that the tissue is fully lysed.

[0033] (2) Centrifuge at 12,000 rpm and 4°C for 5 minutes to remove most of the fat and transfer the clear lysate at the bottom to a new centrifuge tube.

[0034] (3) Add 500 μL of chloroform, shake vigorously manually for 2 minutes, place on ice for 5 minutes, and centrifuge at 12,000 rpm at 4°C for 15 minutes; transfer the supernatant to a new centrifuge tube.

[0035] (4) Add 500 μL of acid phenol chloroform (phenol: chloroform: isoamyl alcohol = 25:24:1), shake vigorously manually for 2 minutes, place on ice for 5 minutes, and centrifuge at 12,000 rpm, 4°C for 10 minutes; remove the supernatant and transfer it to a new centrifuge tube.

[0036] (5) Add 500 μL of isopropanol, invert until thoroughly mixed, and incubate at -20 °C overnight.

[0037] (6) Centrifuge the RNA precipitated overnight at 12,000 rpm and 4°C for 10 minutes, discard the supernatant and retain the precipitate.

[0038] (7) Add 500 μL of pre-chilled 75% ice ethanol to the precipitate, centrifuge at 12,000 rpm, 4 °C for 10 min, and discard the supernatant.

[0039] (8) Repeat the above step (7) to obtain RNA precipitate.

[0040] (9) Place the RNA pellet on a clean bench and blow dry it for 30 minutes to dry the RNA.

[0041] (10) Add an appropriate amount of ultrapure water to the tube containing RNA and incubate at 4°C to allow the RNA to fully dissolve.

[0042] (11) After the RNA is fully dissolved, gently shake to mix, use 1% agarose gel electrophoresis to check the integrity of the RNA, and use NanoDrop 2000 to measure the RNA concentration. Then dilute the RNA to 500 ng / μL with ultrapure water for use.

[0043] 1.3 Reverse transcription

[0044] The total RNA was reverse transcribed into the first cDNA as follows:

[0045] (1) Take 2 μg of total RNA, add water to 8 μL, incubate at 65 ˚C in a PCR instrument for 5 minutes, and quickly cool on ice.

[0046] (2) Add 2 μL of 5× gDNA Buffer, mix gently and centrifuge briefly, incubate at 42 ˚C in a PCR instrument for 3 minutes, and quickly cool on ice.

[0047] (3) Prepare the reaction system mixture as shown in Table 1. The components and amounts are shown in the following table:

[0048] Table 1 Reaction system for reverse transcription into the first cDNA

[0049]

[0050] (4) Add 10 μL of the above mixture to the system in (3), mix gently and centrifuge briefly.

[0051] (5) Incubate at 42 ˚C for 15 minutes and at 95 ˚C for 3 minutes in a PCR instrument, then place on ice to obtain cDNA.

[0052] 1.4 Obtaining the ORF sequence of P120 catenin gene

[0053] Based on the sequence information of P120 catenin protein of known insects, homology comparison was used to search in the transcriptome of locust to obtain the transcript of P120 catenin of locust, and further combined with the genome information of locust to splice a complete P120 catenin gene of locust. The upstream and downstream primer sequences (as shown in SEQ ID No.2 and SEQ ID No.3) were designed using primer premier5.0 software and sent to Beijing Liuhe BGI Gene Technology Co., Ltd. for synthesis.

[0054] The upstream and downstream primer sequences for PCR amplification of P120 catenin gene are as follows:

[0055] P120 catenin-F (SEQ ID No. 2): 5'-ATGGAGCTTTTGTGGCAACC-3';

[0056] P120 catenin-R (SEQ ID No. 3): 5'-TCACACCCATGAGTCACCAC-3'.

[0057] The cDNA reverse transcribed in step 1.3 was used as a template, and the upstream and downstream primers shown in sequences SEQ ID No.2 and SEQ ID No.3 were combined to obtain the ORF fragment of the P120catenin gene by PCR amplification. The fragment was sent to Beijing Liuhe BGI Gene Technology Co., Ltd. for sequencing (all subsequent primer synthesis and sequencing were completed by Beijing Liuhe BGI Gene Technology Co., Ltd.). The ORF sequence of the P120catenin gene is shown in SEQ ID No.1. The full length of the ORF sequence is 963 bp and encodes 320 amino acids.

[0058] 2. Synthetic dsRNA

[0059] 2.1 DNA template for amplification of dsRNA

[0060] Based on the P120 catenin gene sequence shown in SEQ ID No.1, upstream and downstream primers for synthesizing the DNA molecular template sequence of dsP120 catenin (the positive chain is shown in SEQ ID No.6) were designed using primer premier5.0, and the primer sequences are shown in SEQ ID No.4 and SEQ ID No.5, respectively.

[0061] Using the cDNA reverse transcribed in step 1.3 as a template, PCR amplification was performed using the upstream and downstream primers shown in SEQ ID No. 4 and SEQ ID No. 5. The reaction system is shown in Table 2.

[0062] dsP120 catenin-F (SEQ ID No. 4): 5'-TTGCTGGTCAAGTTCTCTTCAC-3';

[0063] dsP120 catenin-R (SEQ ID No. 5): 5'-CATCGTCCTGTCTTCGTATCG-3'.

[0064] Table 2 Reaction system for PCR amplification using cDNA as template

[0065]

[0066] The reaction conditions were as follows: (1) 95°C, 3 min; (2) 95°C, 30 sec; 56°C, 30 sec; 72°C, 30 sec; 35 cycles; (3) 72°C extension for 5 min; and storage at 4°C.

[0067] The single product obtained by PCR amplification was connected to the pGEM-T (Tiangen) vector to obtain the recombinant plasmid pGEM-T-P120 catenin. Sequencing results showed that the size of the PCR amplification product was 203 bp, and its nucleotide sequence was shown in SEQ ID No.6.

[0068] Using the recombinant plasmid pGEM-T-P120 catenin obtained above as a template, dsP120 catenin-T7-F / dsP120 catenin-T7-R (as shown in SEQ ID No.7 and SEQ ID No.8) was amplified by PCR using primers with T7 promoter sequences to obtain a DNA template for synthesizing dsRNA (i.e., the sequence shown in SEQ ID No.6 has T7 promoters at both ends). The PCR reaction system and conditions are the same as those in Table 2. The target fragment was purified using the Wizard® SV Geland PCR Clean-Up System (Promega) kit. The concentration was measured by NanoDrop 2000 and determined to be between 125 ng / μl and 1 mg / μl.

[0069] The upstream and downstream primer sequences of the DNA template for synthesizing dsRNA are the 5' end of the sequence SEQ ID No.4 and the sequence SEQ ID No.5 plus the T7 promoter sequence, as follows (the T7 promoter sequence is underlined):

[0070] dsP120 catenin-T7-F (SEQ ID No. 7):

[0071] 5'- TAATACGACTCACTATAGG TTGCTGGTCAAGTTCTCTTCAC-3′;

[0072] dsP120 catenin-T7-R (SEQ ID No. 8):

[0073] 5'- TAATACGACTCACTATAGG CATCGTCCTGTCTTCGTATCG-3'.

[0074] 2.2 Synthesis of dsRNA

[0075] Using the DNA with T7 promoter sequences at both ends obtained in step 2.1 as a template, use T7RiboMAX TMdsRNA was synthesized by in vitro transcription using the ExpressRNAi System (Promega) kit.

[0076] The in vitro transcription reaction system is shown in Table 3.

[0077] Table 3 In vitro transcription reaction system

[0078]

[0079] The reaction conditions were: 37°C, 60 minutes; 70°C, 10 minutes; 25°C, 30 minutes.

[0080] 2.3 Purification of dsRNA

[0081] (1) Add 160 μL Nuclease-Free water and 200 μL acidic phenol chloroform to the reaction system in step 2.2, mix thoroughly and place on ice for 5 minutes;

[0082] (2) Centrifuge at 13,000 g for 10 min at 4°C and transfer the supernatant into a new 1.5 mL centrifuge tube.

[0083] (3) Add 0.1 volume of sodium acetate (3.0 M, pH 5.2) and 2.5 volume of anhydrous ethanol to the supernatant, mix thoroughly, and place on ice for 5 minutes;

[0084] (4) Centrifuge at 13,000 g for 10 min at 4°C and discard the supernatant.

[0085] (5) Add 500 μL of pre-cooled 75% ethanol, centrifuge at 13,000 g for 10 min at 4°C, and discard the supernatant;

[0086] (6) Repeat step (5) and wash again with 75% ethanol;

[0087] (7) Place in a clean bench to dry for 20 minutes, add 40 μL of Nuclease-Free water, place on ice to dissolve for 20 minutes, mix well, measure the concentration using NanoDrop 2000, and place at -20˚C for later use.

[0088] The obtained dsRNA was sequenced. The sequencing results showed that the dsRNA of the locust P120 catenin gene was a double-stranded RNA consisting of a sense strand and an antisense strand, the nucleotide sequence of the sense strand was shown in SEQ ID No.9, and the nucleotide sequence of the antisense strand was the reverse complementary sequence of SEQ ID No.9.

[0089] Example 2: Experiment on the inhibition of locust egg laying and lethality by P120 catenin gene dsRNA

[0090] 1. Locust dsRNA Injection

[0091] Female locusts within 12 hours after emergence were selected as experimental injection materials and randomly divided into a dsP120 catenin treatment group and a dsGFP control group (locusts injected with dsGFP). 15 μg dsRNA (concentration of about 5 μg / μL, about 3 μL) was injected into the body from the second and third intersegments of the locust abdomen using a 10 μL microinjector. The injected locusts were placed in a well-ventilated metal cage (25 cm × 25 cm × 25 cm) for breeding at a temperature of 30 ± 2 ° C, a photoperiod of 14L:10D, and fed with fresh wheat seedlings twice a day. The mortality of adults was observed and counted every day after injection, and statistics were stopped until the first egg laying.

[0092] 2. P120 catenin gene interference efficiency detection

[0093] 48 hours after dsRNA injection, 8 heads were randomly selected from the dsP120 catenin treatment group and the dsGFP control group, and the fat body and ovary tissues were dissected to detect the interference efficiency of P120 catenin.

[0094] The cDNA of fat body and ovary tissues were obtained according to the method of Example 1. Then, according to the instructions of Tiangen SuperReal Fluorescence Quantitative Premix Reagent Enhanced Edition (SYBR Green) Kit (PF205), qRT-PCR reaction was performed on LightCycler 96 (Roche) using the upstream and downstream primers of P120 catenin as shown in SEQ ID No.10 and SEQ ID No.11 and the upstream and downstream primers of β-actin as shown in SEQ ID No.12 and SEQID No.13, respectively, to detect the relative expression levels of the target gene (P120 catenin) and the internal reference gene (β-actin), and then according to 2 -ΔΔCt The results are shown in Table 1. Figure 1 As shown, compared with the dsGFP control group, the expression of P120 catenin in the dsP120 catenin treatment group decreased by 73.16% and 97.7% in the fat body and ovary, respectively, indicating that P120 catenin was effectively silenced.

[0095] The primer sequences for qRT-PCR reaction are as follows:

[0096] P120 catenin-qRT-F (SEQ ID No. 10): 5'-CGATACGAAGACAGGACGATGC-3';

[0097] P120 catenin-qRT-R (SEQ ID No. 11): 5'-CTGGTTTGGCTAACTGCTGGTA-3'.

[0098] β-actin-qRT-F (SEQ ID No. 12): 5'-AATTACCATTGGTAACGAGCGATT-3';

[0099] β-actin-qRT-R (SEQ ID No. 13): 5'-TGCTTCCATACCCAGGAATGA-3'.

[0100] The qRT-PCR reaction system is shown in Table 4.

[0101] Table 4 qRT-PCR reaction system

[0102]

[0103] The reaction conditions were as follows: (1) pre-denaturation at 95 °C for 10 min; (2) 40 cycles of 95 °C for 10 s, 58 °C for 30 s, and 72 °C for 30 s; (3) melting curve analysis, 95 °C for 60 s, 65 °C for 30 s, and 95 °C for 30 s.

[0104] 3. Statistics of locust egg laying and mortality after dsRNA injection

[0105] Each group was injected with dsGFP, 17 heads, and dsP120 catenin, 21 heads. Each group was repeated 3 times. The survival rate of the worms was counted every day until the female worms laid eggs for the first time. The results are shown in Tables 5, 6 and Figure 2 shown.

[0106] Table 5 Statistical results of the survival rate of migratory locusts after injection of dsRNA

[0107]

[0108] Table 6 Statistical table of the proportion of locusts laying eggs after injection of dsRNA

[0109]

[0110] From Tables 5, 6 and Figure 2It can be seen that the control group injected with dsGFP began to lay eggs on the 7th day after injection, and concentrated on laying eggs within 7-9 days to complete the first egg-laying cycle. There was no significant difference between its egg-laying situation and that of the normally developed female adults, with a 100% egg-laying locust ratio and a 100% survival rate. In the treatment group injected with dsP120 catenin, death occurred on the 1st to 2nd day after injection, and no eggs were laid. Only 1-2 locusts survived on the 16th day, with a survival rate of 98.4% and a 0% egg-laying locust ratio.

[0111] Note: Because only the first egg-laying of migratory locusts is counted, the “-” in the table means that it will no longer be counted.

[0112] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. Application of biological materials for regulating the expression of P120 catenin gene in locust control, characterized in that: The nucleotide sequence of the P120 catenin gene is shown in SEQ ID No. 1; the migratory locust is a female adult migratory locust; The biological material for regulating the expression of the P120 catenin gene is a biological material for inhibiting or reducing the expression of the P120 catenin gene.

2. The use of the biological material for regulating the expression of the P120 catenin gene according to claim 1 in locust control, characterized in that: The biological material is an RNA molecule that inhibits or reduces the expression of the P120 catenin gene.

3. The use of the biological material for regulating the expression of the P120 catenin gene according to claim 2 in the control of migratory locusts, characterized in that: The RNA molecule is a double-stranded RNA molecule, comprising a sense strand nucleotide as shown in SEQ ID No.9 and an antisense strand nucleotide that is reverse complementary to the sequence shown in SEQ ID No.

9.

4. The use of the biological material for regulating the expression of the P120 catenin gene according to claim 1 in locust control, characterized in that: The biological material is a DNA molecule that produces an RNA molecule; the RNA molecule is a double-stranded RNA molecule, comprising a sense strand nucleotide as shown in SEQ ID No.9 and an antisense strand nucleotide that is reverse complementary to the sequence shown in SEQ ID No.

9.

5. The use of the biological material for regulating the expression of the P120 catenin gene according to claim 4 in locust control, characterized in that: The sequence of the DNA molecule is shown as SEQ ID No.

6.

6. A method for preventing female migratory locusts from laying eggs and / or causing them to die, characterized in that: The method comprises treating female adults of migratory locusts with the double-stranded RNA molecule described in claim 3.

7. The method for preventing female migratory locusts from laying eggs and / or causing death according to claim 6, characterized in that: The female adult migratory locust is a female adult migratory locust that has emerged within 12 hours.

8. The method for preventing female migratory locusts from laying eggs and / or causing death according to claim 7, characterized in that: The dosage of the double-stranded RNA molecule is 15 μg per mouse.

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