CN116250548A - Application of Pharyngeal Inhibitory Lateral Body Neuropeptide in Controlling Periplaneta americana - Google Patents

Abstract

The invention discloses the application of the pharyngeal-suppressing lateral body neuropeptide in preventing and treating Periplaneta americana, wherein AST includes AST1 (SEQ ID NO.1) and AST2 (SEQ ID NO.2). The present invention finds that injecting AST1 and AST2 into female adults of Periplaneta americana in the first reproductive cycle can inhibit the synthesis of vitellogenin Vg, and by inhibiting the synthesis of JH in the lateral body of the pharynx, thereby inhibiting the JH signal of the fat body, thereby inhibiting the fat The vitellogenesis of the body can finally inhibit the maturation of the ovary; the purpose of preventing and controlling the American cockroach can be achieved by controlling the reproduction of the female. This method has high specificity, can directly target the reproduction of female Periplaneta americana, does not produce harmful substances that pollute the environment, and provides a new method for pest control.

Description

Application of Pharyngeal Inhibitory Lateral Body Neuropeptide in Controlling Periplaneta americana

technical field

The invention belongs to the technical field of biological control, and in particular relates to the application of the pharyngeal lateral body neuropeptide in preventing and treating Periplaneta americana.

Background technique

Female insect reproduction has received extensive attention due to its importance in species reproduction and its great potential as a target for pest control. Insects are the most abundant and diverse species in the world, with more than 1 million known species. Insects are diverse and highly adaptable, and can be found in activities from the hot tropics to the cold polar regions. Insects are so widely distributed, in addition to their strong adaptability, an important reason is their strong reproductive ability. In most insects, including incomplete metamorphosis, reproduction is dominated by JH, which acts as a gonadotropin to regulate the maturation of reproductive organs in these insects (Roy et al., 2018). In the insect adult stage, JH binds to the receptor Met, activates the expression of its primary response gene Kr-h1, induces the synthesis of Vg in the fat body, promotes the absorption of Vg by the ovary and the development of follicular cells, thereby playing a role in promoting reproduction. features (Roy et al., 2018; Zhu et al., 2020).

The American cockroach (Periplaneta americana) belongs to the order Blatata, and it is an insect of the Blatidae family, commonly known as cockroach. It can transmit pathogenic factors such as bacteria, protozoa and viruses; act as an allergen to trigger allergic reactions in the human body. With the global warming, the acceleration of urbanization, the rapid development of transportation and trade, the degree of harm is becoming more and more serious, and it has become an important sanitary pest in hotels, restaurants, families, hospitals, schools, food processing, manufacturing and sales, catering and other units. Periplaneta americana is an insect with incomplete metamorphosis. It has strong fecundity and environmental adaptability, can carry out parthenogenesis, and spreads pathogenic microorganisms. It is a serious urban sanitation pest. At present, chemical control is still the main way to control the population of Periplaneta americana. The long-term abuse of organophosphorus, carbamate and pyrethroid chemical insecticides will inevitably lead to the improvement of pest resistance and environmental pollution. Therefore, it is imperative to establish a safe and effective cockroach control mechanism.

Contents of the invention

The object of the present invention is to provide the application of AST1 and AST2 proteins in preventing and treating Periplaneta americana.

The technical scheme that the present invention takes is:

The first aspect of the present invention provides the application of a protein in at least one of the following A1) to A14); the amino acid sequence of the protein is shown in SEQ ID NO.1 and/or SEQ ID NO.2;

A1) suppressing the expression of insect vitellogenin gene Vg1 and/or Vg2;

A2) preparing products that inhibit the expression of insect vitellogenin gene Vg1 and/or Vg2;

A3) inhibit insect ovary maturation;

A4) preparing products that inhibit the maturation of insect ovaries;

A5) inhibit insect ovary development;

A6) preparing products that inhibit the development of insect ovaries;

A7) inhibit the development of insect ovarian follicle cells;

A8) preparing a product that inhibits the development of insect ovarian follicle cells;

A9) inhibit the expression of juvenile hormone;

A8) preparing a product that inhibits the expression of juvenile hormone;

A9) inhibit the reproduction of insect females;

A10) preparing a product that inhibits the reproduction of insect females;

A11) pest control;

A12) preparing products for controlling pests;

A13) reduce insect reproductive capacity;

A14) Preparation of products that reduce the reproductive capacity of insects.

The second aspect of the present invention provides the application of the substance that increases the content and/or activity of the protein described in the first aspect of the present invention in at least one of A1) to A14);

A1) suppressing the expression of insect fat body vitellogenin gene Vg1 and/or Vg2;

A2) preparing a product that inhibits the expression of insect fat body vitellogenin gene Vg1 and/or Vg2;

A3) inhibit insect ovary maturation;

A4) preparing products that inhibit the maturation of insect ovaries;

A5) inhibit insect ovary development;

A6) preparing products that inhibit the development of insect ovaries;

A7) inhibit the development of insect ovarian follicle cells;

A8) preparing a product that inhibits the development of insect ovarian follicle cells;

A9) inhibit the expression of juvenile hormone;

A8) preparing a product that inhibits the expression of juvenile hormone;

A9) inhibit the reproduction of insect females;

A10) preparing a product that inhibits the reproduction of insect females;

A11) pest control;

A12) preparing products for controlling pests;

A13) reduce insect reproductive capacity;

A14) Preparation of products that reduce the reproductive capacity of insects.

The third aspect of the present invention provides the application of the substance that improves the expression of the gene encoding the protein described in the first aspect of the present invention in at least one of A1) to A14);

A1) suppressing the expression of insect fat body vitellogenin gene Vg1 and/or Vg2;

A2) preparing a product that inhibits the expression of insect fat body vitellogenin gene Vg1 and/or Vg2;

A3) inhibit insect ovary maturation;

A4) preparing products that inhibit the maturation of insect ovaries;

A5) inhibit insect ovary development;

A6) preparing products that inhibit the development of insect ovaries;

A7) inhibit the development of insect ovarian follicle cells;

A8) preparing a product that inhibits the development of insect ovarian follicle cells;

A9) inhibit the expression of juvenile hormone;

A8) preparing a product that inhibits the expression of juvenile hormone;

A9) inhibit the reproduction of insect females;

A10) preparing a product that inhibits the reproduction of insect females;

A11) pest control;

A12) preparing products for controlling pests;

A13) reduce insect reproductive capacity;

A14) Preparation of products that reduce the reproductive capacity of insects.

In some embodiments of the present invention, the substance that increases the expression of the gene, the substance that increases the activity of the protein, or the substance that increases the content of the protein is the protein described in the first aspect of the present invention and/or is related to the present invention. The protein-related biological material described in the first aspect of the invention, the biological material is any one of the following B1) to B4):

B1) a nucleic acid molecule encoding the protein described in the first aspect of the present invention;

B2) an expression cassette containing the nucleic acid molecule of B1);

B3) a recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B1);

B4) A recombinant microorganism containing the nucleic acid molecule described in B1), or a recombinant microorganism containing the expression cassette described in B2), or a recombinant microorganism containing the recombinant vector described in B3).

The fourth aspect of the present invention provides a method, including increasing the content and/or activity of the protein described in the first aspect of the present invention in insects; the method is used in at least one of C1) to C8):

C1) suppressing the expression of insect fat body vitellogenin gene Vg1 and/or Vg2;

C2) inhibit insect ovary maturation;

C3) inhibit insect ovary development;

C4) inhibit the development of insect ovarian follicle cells;

C5) inhibit the expression of juvenile hormone;

C6) Inhibit the reproduction of insect females;

C7) pest control;

C8) Reduction of insect reproductive capacity.

In some embodiments of the present invention, the method for increasing the content and/or activity of the protein described in the first aspect of the present invention in insects is to use the protein described in the first aspect of the present invention or the protein described in the first aspect of the present invention Relevant biomaterials are introduced into insects.

In some embodiments of the present invention, the biological material is any one of the following B1) to B4):

B1) a nucleic acid molecule encoding the protein described in the first aspect of the present invention;

B2) an expression cassette containing the nucleic acid molecule of B1);

B3) a recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B1);

B4) A recombinant microorganism containing the nucleic acid molecule described in B1), or a recombinant microorganism containing the expression cassette described in B2), or a recombinant microorganism containing the recombinant vector described in B3).

In some embodiments of the invention, the introduction includes injection.

In some embodiments of the invention, the insect is a female adult in the first reproductive cycle.

The fifth aspect of the present invention provides the application of the method described in the fourth aspect of the present invention in controlling pests.

In some embodiments of the present invention, the pest or insect is Periplaneta americana.

The sixth aspect of the present invention provides a composition comprising D1) and D2);

D1) The protein whose sequence is shown in SEQ ID NO.1 or the protein-related biological material shown in SEQ ID NO.1;

D2) The protein whose sequence is shown in SEQ ID NO.1 or the biological material related to the protein shown in SEQ ID NO.1.

In some embodiments of the present invention, the relevant biological material is any one of the following B1) to B4):

B1) a nucleic acid molecule encoding said protein;

B2) an expression cassette containing the nucleic acid molecule of B1);

B3) a recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B1);

B4) A recombinant microorganism containing the nucleic acid molecule described in B1), or a recombinant microorganism containing the expression cassette described in B2), or a recombinant microorganism containing the recombinant vector described in B3).

The beneficial effects of the present invention are:

The present invention finds that injecting AST polypeptides AST1 and AST2 into female adults of Periplaneta americana in the first reproductive cycle can inhibit the synthesis of vitellogenin Vg, and inhibit the JH signal of the fat body by inhibiting the synthesis of JH in the pharyngeal lateral body, and then Inhibit the vitellogenesis of the fat body, and finally inhibit the maturation of the ovary; it can control the reproduction of females, so as to achieve the purpose of preventing and controlling Periplaneta americana. This method has high specificity, can directly target the reproduction of female Periplaneta americana, does not produce harmful substances that pollute the environment, and provides a new method for pest control.

Description of drawings

Figure 1 shows the detection results of the expression levels of fat body vitellogenin genes Vg1 and Vg2 after AST injection; Figure 1A shows the injection of AST1, and Figure 1B shows the injection of AST2.

Figure 2 shows the changes in ovarian morphology after AST injection; Figure 2A shows the changes in ovarian phenotype after AST injection, and Figure 2B shows the changes in the length of the first egg in the ovary after AST injection.

Figure 3 is the change of Vg in the ovary after AST injection; Figure 3A is the change of the total protein content of the ovary after AST injection, Figure 3B is the characteristic sequence of Vg1 identified by mass spectrometry analysis, and Figure 3C is the characteristic sequence of Vg2 identified by mass spectrometry analysis.

Figure 4 shows the morphological changes of follicular cells and Patency after AST injection.

Figure 5 shows the changes of JH synthesis and JH signal after AST injection; Figure 5A shows the injection of AST1, and Figure 5B shows the injection of AST2.

Among them, ***, P<0.001; **, P<0.01; *, P<0.05.

Detailed ways

The conception and technical effects of the present invention will be clearly and completely described below in conjunction with the embodiments, so as to fully understand the purpose, features and effects of the present invention. Apparently, the described embodiments are only some of the embodiments of the present invention, rather than all of them. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative efforts belong to The protection scope of the present invention.

Design and synthesis of embodiment 1 AST

The precursor protein of allatostatin (AST) needs to be cleaved into short mature polypeptides to exert corresponding functions. Based on the AST precursor sequence of Periplaneta americana (NCBI database ID: CAA62500.1), mature AST sequences (SEQ ID No.1 and SEQ ID No.2) were designed. Submit the AST precursor protein sequence to the online neuropeptide analysis and prediction software http://stagbeetle.animal.uiuc.edu/cgi-bin/neuropred.py to predict the signal peptide and enzyme cleavage site. According to the sequence between the enzyme cleavage sites, the mature neuropeptide sequence after cleavage is determined. Mature AST peptides (SEQ ID No.1 and SEQ ID No.2) are synthesized from the C-terminus to the N-terminus using a linear peptide synthesis method (Aiji Biotech, Guangzhou, China).

Periplaneta americana mature AST polypeptide sequence 1 (SEQ ID No.1): SPSGMQRLYGFGL-NH ₂ ;

Periplaneta americana mature AST polypeptide sequence 2 (SEQ ID No. 2): ADGRLYAFGL-NH ₂ .

Example 2 Research on the Effect of AST on Inhibiting the Reproduction of Periplaneta americana Females

(1) AST injection and phenotype observation statistics

Dilute AST1 and AST2 with sterilized ultrapure water to a working solution of 50 μg/μL. Pick healthy female Periplaneta americana on the second day after eclosion, fully anesthetize it with carbon dioxide, inject AST1 and AST2 into the second body segment of the abdomen of Periplaneta americana respectively with a microinjector, and inject 100 μg (2 μL, 50 μg/μL). The control group was injected with sterilized ultrapure water. Injected on the 2nd and 4th day, respectively, a total of 2 injections. Among them, 30 rats were injected for each treatment. On the 5th day, the head and fat body were collected separately, and the tissues of 4 Periplaneta americana were combined into one sample, which was quickly frozen in liquid nitrogen, and then stored at -80°C for future use for total RNA extraction. Four groups were taken for each treatment Samples: Take the ovaries, combine the tissues of 4 Periplaneta americana into one sample, put them into liquid nitrogen for quick freezing, and then store them at -80°C for future use for total protein extraction. Four groups of samples are taken for each treatment; the ovaries are taken for Photographing, DAPI and Phalloidin staining and confocal microscope photographing, respectively taking pictures of four pairs of ovaries. The experiment was repeated three times.

(2) Effect of AST on the expression of fat body vitellogenin genes Vg1 and Vg2

According to the gene sequence in the American cockroach genome database (Li et al., 2018), the primer design software Primer5 was used to design fluorescent quantitative PCR primers, and the length of the amplified fragment was about 80-150bp. Primers used in fluorescent quantitative PCR detection:

Vg1-FP: TGCTGATGAGGACACAACCT (SEQ ID No. 3);

Vg1-RP: CCACTTGCTTCACTGGATCG (SEQ ID No. 4);

Vg2-FP: GAGAAGCAGACACGAGGAACG (SEQ ID No. 5);

Vg2-RP: CCCTTGAACGCCACTGAGAC (SEQ ID No. 6);

actin-FP: CATCCTGCGTTTGGATCTGG (SEQ ID No. 7);

actin-RP: TTTCTCGTTCGGCAGTGGTG (SEQ ID No. 8).

Green Master Mix(Low Rox Plus)试剂，按照其说明书操作，进行荧光定量PCR检测。两组数据之间进行比较，利用管家基因actin对目的基因相对表达量进行计算，再使用t检验比较两组数据之间是否存在显著差异，得出目的基因在某处理或者某时间点的相对表达量。每个样品重复三次，每个处理进行三次生物学重复，取各组数据的平均值和标准误，绘制图表。Using HieffTM qPCR

The Green Master Mix (Low Rox Plus) reagent was operated according to its instruction manual for fluorescent quantitative PCR detection. Compare the two sets of data, use the housekeeping gene actin to calculate the relative expression of the target gene, and then use the t test to compare whether there is a significant difference between the two sets of data, and obtain the relative expression of the target gene in a certain treatment or a certain time point quantity. Each sample was repeated three times, and each treatment was carried out three biological repetitions, and the average value and standard error of each group of data were taken, and a graph was drawn.

After AST1 injection, the expression levels in the fat body were detected, and it was found that the expression levels of Vg1 and Vg2 were significantly down-regulated by more than 80% (Figure 1A); after AST2 injection, the expression levels of Vg1 and Vg2 were also significantly down-regulated by nearly 80% (Figure 1B). These results indicate that AST represses the expression of fat body vitellogenin genes Vg1 and Vg2.

(3) Ovarian phenotype observation

After injection of AST1 and AST2, take out the Periplaneta americana, fix it on a wax plate with an insect needle after anesthesia, place it under an Olympus SZ61 dissecting microscope, cut open the abdomen, peel off the ovary from the tissue, take it out and put it in a petri dish, add the American cockroach Periplaneta physiological saline to make the ovaries float in the solution. Continue to dissect the ovary under a microscope, take out the peripheral muscle tissue of the ovary, and take pictures with a Nikon DS-Ri2 camera to record the development of the ovary. Under a microscope, the ovaries were stripped into individual tubules with dissecting forceps, the shape of the tubules was observed, photographed and recorded with a Nikon DS-Ri2 camera, and the length of the first egg in the ovary was measured using NIS-Elements BR4.50.00 software.

Ovarian development was inhibited after AST1 and AST2 injections (Fig. 2A), and the first egg length was significantly reduced by 63% and 61.4%, respectively, compared with the control (Fig. 2B). The above results indicated that ovarian maturation was inhibited after AST injection, indicating that AST had a significant inhibitory effect on oogenesis.

(4) Effect of AST on total ovarian protein

After treatment, the Periplaneta americana ovary was ground in Beyontian RIPA lysate (PMSF was added in advance, the final concentration was 1mM), centrifuged at 12000g for 30min at 4°C, and the supernatant was filtered through a 0.22μm filter membrane. The protein concentration was determined using Yisheng Bio BCA kit, and the concentration was adjusted to 2ug/ul. Add Loading buffer to the protein sample and put it in a boiling water bath for 5 minutes; the sample volume of each gel hole is 10ul; during electrophoresis, first use 80V voltage for 30min, after the sample enters the separation gel, adjust to 130V and continue electrophoresis for 90min; remove the gel and put it in Put in the staining dish with Coomassie Brilliant Blue R250 staining solution, place it on a shaker, set the speed at 45r/min, pour out the dye solution after 30 minutes of staining, rinse with water 3 times; add decolorization solution, place it on the shaker, the bands are clearly visible Finally pour off the decolorization solution; put it in the scanner and take pictures.

According to the results of SDS-PAGE gel electrophoresis, after the injection of AST, the total protein content of the ovary decreased significantly (Fig. 3A). Among them, there is an obvious protein band at 100kD, and the band is subjected to gel cutting mass spectrometry detection and analysis, and it is found that the band is Vg, including Vg1 and Vg2, and its characteristic polypeptide sequence is EPGNLNLAR (SEQ ID No.9) (Fig. 3B) and HELIAVAYALPSK (SEQ ID No. 10) (Fig. 3C). The above results show that AST inhibits the absorption of Vg protein by the ovary, thereby inhibiting the accumulation of Vg in the ovary and preventing the development of the ovary.

(5) Effect of AST on the development of ovarian follicle cells

Take out the Periplaneta americana on the 5th day after treatment, dissect out the ovary after anesthesia, put it into a 0.5mL centrifuge tube, add 400ul fixative solution, and fix it on a shaker at 25°C for 1 hour; suck out the fixative solution, add 400μL PBT to wash it, and pour it out Wash the waste liquid repeatedly 4 times; add 400ul PBT, and place it on a shaker at 25°C for fine washing for 1 hour; suck out the PBT, add 400μL of PBT again, and add 1ul DAPI/phalloidin (1:10000), wrap the centrifuge tube with tin foil to avoid Stain on a shaker for 30 minutes; then wash with PBT 4 times, 400 μL each time; add 400 μL of PBT, and place on a shaker at 25°C for fine washing for 1 hour; conduct a second dissection, take out the tissue and put it on a glass slide, and blot dry Liquid, drop a certain amount of 50% glycerol, cut out the desired tissue, mount the slide, observe and take pictures under a confocal microscope. Photographs of ovarian follicle cells were randomly selected under a confocal microscope with a 40× objective lens and taken at a magnification of 5 times. Three photos were randomly selected for each treatment.

After AST injection, the development of ovarian follicle cells was inhibited (Fig. 4). Compared with the control group, the intercellular patency (white arrow) in the AST1 and AST2 treatment groups disappeared, the development of follicle cells was inhibited, and the cells were significantly smaller. The results showed that AST inhibited the development of follicular cells and the formation of patency.

(6) Detection of JH key enzyme gene expression in pharyngeal body and JH signal transduction gene expression in fat body

After AST1 and AST2 treatment, the expressions of key juvenile hormone JH synthase genes Jhamt and Cyp15a1 in the lateral body of the pharynx were significantly reduced (Fig. 5A), and the expressions of key juvenile hormone JH signal transduction genes Met and Kr-h1 in the fat body also significantly decreased (Fig. 5B). These results suggest that AST inhibits JH signaling in the fat body by inhibiting JH synthesis in the pharyngeal lateral body.

Based on the previous experimental results in Figure 1-Figure 5, the experiment shows that AST inhibits the JH signal of the fat body by inhibiting the synthesis of JH in the lateral body of the pharynx, thereby inhibiting the vitellogenesis of the fat body, and finally inhibiting ovarian maturation. Experiments have proved that AST has the function of inhibiting the female reproduction of Periplaneta americana. By controlling the reproduction of females, the effect of preventing and controlling Periplaneta americana can be achieved.

The above-mentioned specific embodiments have described the present invention in detail, but the present invention is not limited to the above-mentioned embodiments, and various changes can be made within the knowledge of those of ordinary skill in the art without departing from the gist of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other if there is no conflict.

Claims (10)

1. The use of a protein in at least one of the following A1) to A14); the amino acid sequence of the protein is shown as SEQ ID NO.1 and/or SEQ ID NO. 2;

a1 Inhibiting expression of insect vitellogenin genes Vg1 and/or Vg 2;

a2 Preparing a product for inhibiting the expression of the insect vitellogenin genes Vg1 and/or Vg 2;

a3 Inhibiting insect ovarian maturation;

a4 Preparing a product for inhibiting the maturation of the ovaries of the insects;

a5 Inhibiting insect ovarian development;

a6 Preparing a product for inhibiting the development of insect ovaries;

a7 Inhibiting insect ovarian follicular cell development;

a8 Preparing a product for inhibiting development of ovarian follicular cells of the insect;

a9 Inhibiting expression of juvenile hormone;

a8 Preparing a product that inhibits juvenile hormone expression;

a9 Inhibiting female reproduction of insects;

a10 Preparing a product for inhibiting reproduction of female insects;

a11 A) controlling pests;

a12 Preparing a product for controlling pests;

a13 Reducing the reproductive capacity of insects;

a14 A product that reduces the reproductive capacity of insects is prepared.

2. Use of a substance which increases the protein content and/or activity of claim 1 in at least one of A1) to a 14);

a1 Inhibiting expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a2 Preparing a product for inhibiting the expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a3 Inhibiting insect ovarian maturation;

a4 Preparing a product for inhibiting the maturation of the ovaries of the insects;

a5 Inhibiting insect ovarian development;

a6 Preparing a product for inhibiting the development of insect ovaries;

a7 Inhibiting insect ovarian follicular cell development;

a8 Preparing a product for inhibiting development of ovarian follicular cells of the insect;

a9 Inhibiting expression of juvenile hormone;

a8 Preparing a product that inhibits juvenile hormone expression;

a9 Inhibiting female reproduction of insects;

a10 Preparing a product for inhibiting reproduction of female insects;

a11 A) controlling pests;

a12 Preparing a product for controlling pests;

a13 Reducing the reproductive capacity of insects;

a14 A product that reduces the reproductive capacity of insects is prepared.

3. Use of a substance which increases the expression of a gene encoding a protein according to claim 1 in at least one of A1) to a 14);

a1 Inhibiting expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a2 Preparing a product for inhibiting the expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a3 Inhibiting insect ovarian maturation;

a4 Preparing a product for inhibiting the maturation of the ovaries of the insects;

a5 Inhibiting insect ovarian development;

a6 Preparing a product for inhibiting the development of insect ovaries;

a7 Inhibiting insect ovarian follicular cell development;

a8 Preparing a product for inhibiting development of ovarian follicular cells of the insect;

a9 Inhibiting expression of juvenile hormone;

a8 Preparing a product that inhibits juvenile hormone expression;

a9 Inhibiting female reproduction of insects;

a10 Preparing a product for inhibiting reproduction of female insects;

a11 A) controlling pests;

a12 Preparing a product for controlling pests;

a13 Reducing the reproductive capacity of insects;

a14 A product that reduces the reproductive capacity of insects is prepared.

4. Use according to claim 2 or 3, wherein the substance that increases the expression of the gene, the substance that increases the activity of the protein or the substance that increases the content of the protein is a protein as defined in claim 1 and/or a biological material related to a protein as defined in claim 1, which biological material is any one of the following B1) to B4):

b1 A nucleic acid molecule encoding the protein of claim 1;

b2 An expression cassette comprising the nucleic acid molecule of B1);

b3 A recombinant vector comprising the nucleic acid molecule of B1) or a recombinant vector comprising the expression cassette of B1);

b4 A recombinant microorganism comprising the nucleic acid molecule of B1), a recombinant microorganism comprising the expression cassette of B2), or a recombinant microorganism comprising the recombinant vector of B3).

5. A method comprising increasing the content and/or activity of a protein as defined in claim 1 in an insect, said method being used for at least one of C1) to C8):

c1 Inhibiting expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

c2 Inhibiting insect ovarian maturation;

c3 Inhibiting insect ovarian development;

c4 Inhibiting insect ovarian follicular cell development;

c5 Inhibiting expression of juvenile hormone;

c6 Inhibiting female reproduction of insects;

c7 A) controlling pests;

c8 Reducing the reproductive capacity of insects.

6. The method according to claim 5, wherein the method of increasing the content and/or activity of the protein according to claim 1 in an insect is to introduce the protein according to claim 1 or the protein-related biological material according to claim 1 into the insect.

7. The method according to claim 6, wherein the biological material is any one of the following B1) to B4):

b1 A nucleic acid molecule encoding the protein of claim 1;

b2 An expression cassette comprising the nucleic acid molecule of B1);

b3 A recombinant vector comprising the nucleic acid molecule of B1) or a recombinant vector comprising the expression cassette of B1);

b4 A recombinant microorganism comprising the nucleic acid molecule of B1), a recombinant microorganism comprising the expression cassette of B2), or a recombinant microorganism comprising the recombinant vector of B3).

8. The method of claim 6, wherein the introducing means comprises injection.

9. A composition comprising D1) and D2);

d1 Protein with a sequence shown as SEQ ID NO.1 or protein related biological material shown as SEQ ID NO. 1;

d2 A protein with a sequence shown as SEQ ID NO.1 or a protein related biological material shown as SEQ ID NO. 1.

10. The use according to any one of claims 1 to 4 or the method according to any one of claims 5 to 8, wherein the pest or insect is american cockroach.

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