Method for obtaining polytype male and female interstitial apis cerana

Abstract

The invention provides a method for obtaining polytype male and female interstitial Pingewei, and belongs to the technical field of arthropod reproductive mode and phenotype regulation, and the method comprises the following steps: feeding Pingewei carrying Ricktsia for several days by using a rifampicin solution, and feeding eggs to the Pingewei carrying Ricktsia for 24 + / -1h after feeding the rifampicin solution for parasitism, so as to obtain the polytype male and female interstitial Pingewei, and obtaining the polytype male and female interstitial Pingewei, so as to obtain the polytype male and female interstitial Pingewei, and the polytype male and female interstitial Pingewei. On the second day, the third day and the fourth day after the rifampicin solution is fed, polytypic male and female body surface types appear on the offspring, and the male and female body small bees are obtained. According to the method disclosed by the invention, the polytype male and female interbody Pingketsia is obtained by partially removing a symbiotic bacterium Rickketsia in the Pingketsia, so that a model for interaction between the Rickketsia and the host Pingketsia is favorably established, a foundation is laid for researching sex determination and differentiation of the host, and the research on the origin evolution of the sex type II of the insect and the formation reason of the male and female interbody is facilitated.

Description

Technical Field

[0001] The invention belongs to the technical field of arthropod reproduction mode and phenotype regulation, and particularly relates to a method for obtaining polymorphic intersex flat-abdomen wasps. Background Art

[0002] Hermaphroditism refers to organisms that have both male and female characteristics. In fact, most flowering plants (94%) have both male and female sex organs, usually in the same flower. In contrast, hermaphroditism is rare in the entire animal kingdom (about 5% of all species), mainly concentrated in fish and most snails, corals, trematodes, barnacles and many echinoderms, while hermaphroditism is less reported in species-rich insects. Gynandromorphs and Intersexes are the main types of hermaphroditism in animal studies. Parts that can be clearly attributed to one sex (i.e., have obvious male and female characteristics and clear boundaries) are called Gynandromorphs, and those that express an intermediate phenotype between female and male are called Intersexes. The former may be related to developmental and fertilization errors and are genetic mosaics; the latter are related to factors such as temperature, symbiotic bacterial infection, hormone deviation or loss-of-function mutations. Since hermaphrodites are often rare in animals and most of them are infertile, current research in this area is mostly focused on phenotypic description.

[0003] Some endosymbionts will break the host's sex determination pattern to enhance their own spread. This sex manipulation will produce genetic conflicts, thus forming resistance genes and eventually driving the transformation of the sex determination system. Males are generally a dead end for the spread of symbionts, so some symbionts will induce host parthenogenesis, feminization and male killing. Common symbionts that affect host parthenogenesis include Wolbachia, Rickettsia, and Cardinium. Among arthropods, more than half (52%) of the hosts are infected by Wolbachia, about a quarter (24%) are infected by Rickettsia, and about one eighth (13%) are infected by Cardinium. Studies have shown that Rickettsia can cause apomixis in the host and then cause the host to parthenogenetic. There are now 4 cases of Rickettsia-induced parthenogenesis in Hymenoptera wasps, but there are no reports on studies on Rickettsia-induced intersex bodies in hosts, which account for 1 / 4 of arthropods.

[0004] Anastatus gansuensis (Anastatidae, Anastatus genus) is a new species of parasitic wasp found in Caligula japonica Moore (Lepidoptera: Bombycidae), a major walnut pest in western China. Studies have found that this parasitic wasp mainly reproduces parthenogenetically indoors, and the offspring parasitizing on eggs of Ginkgo moth at different ages and eggs of tussah silkworms in different states are all female. In addition, previous studies have found that Anastatus gansuensis is heavily infected with Rickettsia. If the intersex body of Anastatus gansuensis can be obtained, it will be an excellent model for studying the interaction between symbionts and hosts, especially in terms of sex determination and differentiation. It can lay the foundation for exploring the sex determination and differentiation of the host, and help study the origin and evolution of sexual dimorphism in insects and the reasons for the formation of intersex bodies. Summary of the invention

[0005] In order to obtain intersexual planus, the present invention provides a method for obtaining polymorphic intersexual planus, which obtains polymorphic intersexual planus by partially removing the endosymbiotic bacteria Rickettsia of the planus, which is beneficial to establish a model of interaction between Rickettsia and the host planus, lays a foundation for exploring the sex determination and differentiation of the host, and helps to study the origin and evolution of sexual dimorphism in insects and the reasons for the formation of intersexuality.

[0006] The present invention is achieved through the following technical solutions:

[0007] The present invention provides a method for obtaining a polymorphic intersexed flat-abdomen wasp, the method comprising:

[0008] The flat-bellied wasps carrying Rickettsia were fed with a rifampicin solution for several days, and eggs were given to the flat-bellied wasps carrying Rickettsia for parasitism 24±1h after feeding the rifampicin solution. On the 2nd, 3rd and 4th days after feeding the rifampicin solution, the offspring showed a polymorphic intersex phenotype, thus obtaining intersex flat-bellied wasps.

[0009] Furthermore, the concentration of rifampicin in the rifampicin solution is 0.1 to 10.0 mg / mL.

[0010] Preferably, the rifampicin solution comprises a fructose solution containing rifampicin.

[0011] Alternatively, the present invention provides a method for obtaining a polymorphic intersexed flat-abdomen wasp, the method comprising:

[0012] The adults of the flat-bellied wasp carrying Rickettsia were used as parents;

[0013] Feeding the parent with a rifampicin solution for several days, and administering worm eggs to the parent for parasitism 24±1h after feeding the rifampicin solution;

[0014] On the 2nd, 3rd and 4th days of feeding the rifampicin solution, the offspring showed a polymorphic intersex phenotype, namely, the intersex flat-abdomen wasp was obtained.

[0015] Furthermore, the method of using the adult wasp carrying Rickettsia as a parent specifically includes:

[0016] Adult flat-bellied wasps carrying Rickettsia are used as parents, and the emergence time of the parents is less than 6 hours.

[0017] Further, the step of feeding the parent with a rifampicin solution for several days specifically comprises:

[0018] Feeding the parent with a rifampicin solution having a rifampicin concentration of 0.1 to 10.0 mg / mL for several days;

[0019] Wherein, the rifampicin solution comprises a fructose solution containing rifampicin.

[0020] Furthermore, the step of giving worm eggs to the parent for parasitism 24±1h after feeding the rifampicin solution specifically includes:

[0021] After feeding the rifampicin solution for 24±1h, the parents are given tussah eggs for parasitism, and then fresh tussah eggs are replaced at intervals of 24±1h.

[0022] Furthermore, on the 2nd, 3rd and 4th days of feeding the rifampicin solution, the offspring show a polymorphic intersex phenotype, that is, the intersex flat-abdomen wasp is obtained, specifically comprising:

[0023] On days 2, 3, and 4 of feeding with the rifampicin solution, the offspring showed polymorphic intersex phenotypes;

[0024] Individuals that have both female and male bee characteristics are classified as an intersexual, and the intersexuals are collected to obtain intersexual flat-bellied wasps.

[0025] Based on the same inventive concept, the present invention provides a polymorphic intersexual flat-abdomen wasp, which is constructed by the above method for obtaining a polymorphic intersexual flat-abdomen wasp.

[0026] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0027] 1. The present invention discloses a method for obtaining polymorphic intersexual flat-bellied wasps. The present invention finds for the first time that by feeding low concentrations of rifampicin to the mother generation of flat-bellied wasps daily, intersexuals of various phenotypes can be obtained in their offspring while partially removing the endosymbiotic bacteria Rickettsia. The polymorphic intersexual flat-bellied wasps are beneficial to establishing a model of the interaction between Rickettsia and the host flat-bellied wasps, laying a foundation for exploring the sex determination and differentiation of the host, and contributing to studying the origin and evolution of sexual dimorphism in insects and the causes of intersexual formation.

[0028] 2. The present invention discloses a method for obtaining polymorphic intersexual flat-bellied wasps. The method feeds low concentrations of rifampicin to the mother generation of flat-bellied wasps infected with Rickettsia on a daily basis, so that the Rickettsia in the host body is within a specific titer range. Under specific titer conditions, various types of intersexual offspring can be obtained. The method is time-saving, labor-saving, simple and convenient. Various intersexual phenotypes can be obtained in the first generation, which lays a foundation for further research on Rickettsia regulating the reproductive mode of arthropods, helps to understand the evolution of insect sexual characteristics, enriches the interaction process between symbiotic bacteria and hosts, and is an excellent model for exploring species sex determination and differentiation.

[0029] 3. The present invention discloses a method for obtaining polymorphic intersexuals of flat-abdomen wasps. The method is the first method for obtaining polymorphic intersexuals of flat-abdomen wasps. The inventors found through experiments that after the flat-abdomen wasps were fed with rifampicin, the reproductive mode of the wasps changed from parthenogenetic theriogenesis to parthenogenetic androgenesis, during which various types of intersexuals appeared. During this process, the titer of endophyte Rickettsia gradually decreased, and intersexuals appeared after a specific number of days of antibiotic treatment, at which time the Rickettsia titer was at a medium level. Combining molecular technology and biological verification, the experiment proved that the polymorphic intersexual phenotype of flat-abdomen wasps appeared in a specific Rickettsia titer range (4.34-3.93 gene copies / mic). Above the titer range, normal-shaped female individuals were obtained, and below the titer range, normal-shaped male individuals were obtained. The acquisition of intersexuals provides a good model for studying the interaction between Rickettsia and the host, and also provides good research materials for studying the effects of symbiotic bacteria on host sex determination, differentiation, homology of male and female individuals, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0031] Figure 1Survival analysis of the Gansu flat-bellied wasp fed with different concentrations of rifampicin.

[0032] Figure 2 The time and proportion of the appearance of intersex progeny when the queen bees were treated with 0.1 mg / mL rifampicin-fructose water daily.

[0033] Figure 3 This is a normal female individual of the Gansu flat-bellied wasp.

[0034] Figure 4 This is a normal male individual of the Gansu flat-bellied wasp.

[0035] Figure 5 (AN) shows the sorting of the intersex bodies of Gansu flat-bellied wasps according to the degree of masculinization of the individuals: Figure 5A , Figure 5D , Fig. 5F , Figure 5G , Figure 5H and Figure 5M It is a mixed hermaphrodite; Figure 5B , Figure 5C and Figure 5K It is a left-right intersexual body. Figure 5E , Fig.5I , Figure 5J , Figure 5L and Figure 5N It is an anterior-posterior intersexual body.

[0036] Figure 6 Changes in Rickettsia titer in the ovaries of mother bees during daily feeding with 0.1 mg / mL rifampicin-fructose water. DETAILED DESCRIPTION

[0037] The present invention will be described in detail below in conjunction with specific implementations and examples, and the advantages and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific implementations and examples are used to illustrate the present invention, rather than to limit the present invention.

[0038] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. In the event of a conflict, the present specification takes precedence.

[0039] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0040] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0041] The following is a detailed description of a method for obtaining polymorphic intersex flat-abdomen wasps according to the present invention in combination with examples and experimental data.

[0042] 1. Test insects

[0043] The Gansu flat-bellied wasp used in the experiment was collected from a walnut orchard in Gansu Province, China. The parasitic wasp was identified based on morphological characteristics and further confirmed by COI sequence (GenBank: MK373759). Gansu flat-bellied wasp was raised with tussah silkworm eggs, and the parasitized tussah silkworm eggs were placed in a culture room for eclosion. Through 16S third-generation full-length sequencing, the bee was severely infected with and only with the existing Rickettsia that induces parthenogenesis.

[0044] 2. Reagent Preparation

[0045] 1 mol / L fructose water was used as a solvent to prepare 0.10 mg / mL, 1.00 mg / mL, and 10.00 mg / mL rifampicin fructose water.

[0046] Example 1

[0047] This embodiment provides a method for obtaining a polymorphic intersex flat-abdomen wasp.

[0048] 1. Experimental Methods

[0049] (1) Rifampicin concentration screening: Place newly emerged (<6 hours) Gansu flat-bellied bees in a transparent jar (12.5 cm*6.5 cm) as a replicate, feed them with the same concentration of rifampicin-fructose aqueous solution every day, place them in a culture greenhouse, and observe and record the death of the bees. The control group was only fed with fructose water, and the other conditions were the same as the treatment group.

[0050] (2) Antibiotic feeding and parasitism: The newly emerged Gansu flat-bellied wasps were placed in glass tubes (1.5*10 cm) and fed with the same concentration of rifampicin-fructose water every day and replaced with fresh tussah egg cards every day for parasitism. The parasitized egg cards were placed in the culture room until they emerged. The male-female ratio of the offspring was recorded, and the type and number of male-female abnormal individuals were recorded. The control group was fed with fructose water, and the other conditions were the same as those of the treatment group.

[0051] (3) Classification of intersex bodies: In order to better classify abnormal male and female individuals, normal male and female adult individuals before and after the removal of antibiotics were used as references, and the head, thorax, abdomen, dorsal plate, antennae, legs and wings of the bees were used as the criteria for judging abnormal individuals. They were divided into three categories: left-right type (divided from left to right), front-back type (divided from front to back) and mixed type (a combination of other types).

[0052] (4) Determination of Rickettsia titer: To determine the abundance of Rickettsia in the body of Gansu flat-bellied bee, we used Premix Ex Taq TM (Takara) and Bio-Rad CFX96 TM The measurement was performed using a Real-Time System (BioRad).

[0053] DNA was extracted from the Gansu flat-bellied wasps fed with 30% honey water, and the copy number of gltA gene was quantified using Rickettsia specific primers.

[0054] The DNA of Gansu flat-bellied wasp was extracted using a kit (TIANamp Genomic DNA kit), and the specific operations were carried out according to the instructions of the kit.

[0055] PCR was used to detect the infection of Rickettsia in Gansu flat-bellied bees. A pair of primers targeting the gltA gene sequence of Rickettsia were used: gltA-F and gltA-R. The sequence is:

[0056] gltA-F:5'-GCTCTTCTCATCCTATGGCTATTAT-3'

[0057] gltA-R: 5'-CAGGGTCTTCGTGCATTTCTT-3'.

[0058] Reaction system:

[0059] Reaction solution components Usage <![CDATA[ddH2O]]> 8.5μL PremixTaq(TaKaRaTaqTMVersionplusdye) 12.5μL gltA-F 1μL gltA-R 1μL Template DNA 2μL

[0060] Total volume 25 μL

[0061] Amplification conditions:

[0062] Pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 53.7°C for 30 s, extension at 72°C for 30 s, 35 cycles; extension at 72°C for another 10 min, and cycle at 4°C.

[0063] The PCR products were detected by 2% agarose gel electrophoresis, and 2kb Marker was used as a control. 5μL of PCR products and Marker were added to 2% agarose gel and immediately placed in l*TAE buffer for electrophoresis detection. The electrophoresis conditions were voltage 120V and time 20min.

[0064] The steps of gel recovery and purification of DNA fragments, ligation of target gene and vector, heat shock transformation of Escherichia coli competent cells, positive clone screening and sequencing, and plasmid extraction were all performed according to the instructions of the kit.

[0065] Quantitative PCR detection

[0066] The standard curve was drawn by the standard plasmid samples extracted above, and the concentrations were 10 4 , 10 5 , 10 6 , 10 7 and 10 8 Gene copy number / mic.

[0067] The ovaries of mother bees at time points 1, 3, 5, 7, and 10 days were dissected, with 20 repetitions at each time point. DNA was extracted from the ovaries for absolute quantification. After DNA extraction, the abundance of Rickettsia in the bees under different treatments was determined by absolute quantification.

[0068] Reaction system:

[0069] Reaction solution components Usage 2×TBGreenPremixExTaqII 10.0μL Forward primer (10 μmol / L) 0.8μL Reverse primer (10 μmol / L) 0.8μL TemplateDNA 2.0μL <![CDATA[ddH2OUpto]]> 20μL

[0070] Primer sequences:

[0071] glt375-F:5'-TGGTATTGCATCGCTTTGGG-3'

[0072] glt574-R:5'-TTTCTTTAAGCACTGCAGCACG-3'.

[0073] Total volume 20 μL

[0074] Amplification conditions:

[0075] Pre-denaturation at 95℃ for 30s; denaturation at 95℃ for 5s, extension at 60℃ for 30s, 40 cycles; denaturation at 95℃ for 10s; melting curve from 65℃ to 95℃. The temperature was increased every 5s, and the temperature was increased by 0.5℃ each time.

[0076] 2. Experimental Results

[0077] 1. Feeding with low concentration of rifampicin-fructose water had no significant effect on the mortality rate of bees.

[0078] like Figure 1 (R: represents rifampicin, ***p<0.001) shows that compared with the control CK, high concentration of rifampicin solution (1 mg / mL, 10 mg / mL) has a significant toxic effect on the bees, while low concentration of rifampicin solution (0.1 mg / mL) has no significant effect on the mortality rate of the bees.

[0079] Therefore, 0.1 mg / mL is preferred when selecting the antibiotic concentration.

[0080] 2. Antibiotic-treated mother bees obtain intersex bodies at specific times.

[0081] like Figure 2 As shown in the figure, the offspring intersex bodies are mainly concentrated in the 2nd, 3rd, and 4th days of the antibiotic-treated mother bees. On the first day of treatment, the offspring are completely female individuals, and after 5 days of treatment, the offspring will appear completely male individuals. Intersex bodies appear in a transitional period.

[0082] 3. Classification of intersex bodies

[0083] According to references, abnormal individuals are mainly divided into 3 categories and 14 types. The first category is the mixed type, accounting for 6 / 14; the second category is the left-right type, accounting for 3 / 14; the third category is the anterior-posterior type, accounting for 5 / 14, and the mosaic parts are irregularly distributed in the tissue.

[0084] Figure 3 (A)-(I), Figure 4 (A)-(I) show various parts of normal female and male individuals of Gansu flat-bellied wasp, which are used for comparison with the various parts of the abnormal individuals in Figure 5.

[0085] The first type is a mixed hermaphrodite ( Figure 5A , D, F, G, H, M). The first example has a female right wing, a male left wing, and a mixed dorsal plate ( Figure 5A ). The second case has a completely male head with two male antennae and a male foot (right foot) ( Figure 5D ). The third example shows a male wing (left), except for the right front dorsal plate which has female characteristics (right), the rest of the dorsal plate is male, and the abdomen tends to be male ( Fig. 5F ). The fourth case has a completely male head with two male antennae, two male forelegs, a male left wing, and a right wing that has not been fully transformed into a transparent wing ( Figure 5G ). The fifth case is more similar to males overall ( Figure 5H ), a complete male dorsal plate, 2 male wings, 3 male legs (right) and 1 male antennae (left). In the last case, it has female characteristics on the left antennae and right foreleg, and all male characteristics on the rest of the body ( Figure 5M ).

[0086] The second type of left and right type ( Figure 5B, C, K), this category is often accompanied by inconsistent wings, and differences in feet and back plates. Because of the differences between male and female individuals, the differences in size and color can well analyze the differences in details of abnormal individuals. The back plates of female individuals have a metallic color glow, while males do not; female wings have shapes and colors, while male wings are transparent; female feet are in line with the size of the body, often thick and brown, while male feet are small and yellow.

[0087] The third type of front and back type ( Figure 5E , I, J, L, N), including those with male characteristics only in the abdomen and female characteristics in the rest of the body (1 / 5) ( Figure 5E ). Two-fifths of the individuals have male heads and partially or completely female bodies ( Fig.5I ,J), while individuals with a complete female head and the rest of the body showing male characteristics account for only 1 / 5 ( Figure 5L The remaining one retained female characteristics in the abdomen, with the rest of the body showing male characteristics ( Figure 5N ).

[0088] 4. Partial elimination of Rickettsia leads to polymorphic intersex.

[0089] The Rickettsia titer in the ovaries of queen bees was measured by quantitative PCR on the 1st, 3rd, 5th, 7th and 10th days of daily antibiotic treatment. Figure 6 As shown, the results showed that the Rickettsia titer in the ovary was gradually decreasing. The content of Rickettsia was lower than that of the control group after the first day of feeding, and only higher than that of the control group on the first day. At the time of the appearance of intersex bodies (the third day), the specific titer in the ovary (4.10±0.00 copies / mic) was in the middle level between the offspring with all females (4.34±0.00 copies / mic) and the offspring with all males (3.93±0.00 copies / mic) ( Figure 6 ), which is consistent with the previous observation that the offspring gradually transformed from normal female individuals to male individuals with antibiotic treatment, and that intersex bodies appeared on specific days (2, 3, and 4 days) ( Figure 2 ).

[0090] Finally, it should be noted that the terms "comprises," "includes," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements that are inherent to such process, method, article, or apparatus.

[0091] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0092] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A method for obtaining a polymorphic intersex flat-abdomen wasp, characterized in that: The method comprises: The flat-bellied wasps carrying Rickettsia were fed with a rifampicin solution for several days, and eggs were given to the flat-bellied wasps carrying Rickettsia for parasitism 24±1h after feeding the rifampicin solution. On the 2nd, 3rd and 4th days after feeding the rifampicin solution, the offspring showed a polymorphic intersex phenotype, thus obtaining intersex flat-bellied wasps.

2. The method for obtaining polymorphic intersexed flat-abdomen wasps according to claim 1, characterized in that: The concentration of rifampicin in the rifampicin solution is 0.1-10.0 mg / mL.

3. The method for obtaining polymorphic intersexed flat-abdomen wasps according to claim 1, characterized in that: The rifampicin solution comprises a fructose solution containing rifampicin.

4. The method for obtaining polymorphic intersexed flat-abdomen wasps according to claim 1, characterized in that: The method comprises: The adults of the flat-bellied wasp carrying Rickettsia were used as parents; Feeding the parent with a rifampicin solution for several days, and administering worm eggs to the parent for parasitism 24±1h after feeding the rifampicin solution; On the 2nd, 3rd and 4th days of feeding the rifampicin solution, the offspring showed a polymorphic intersex phenotype, namely, the intersex flat-abdomen wasp was obtained.

5. The method for obtaining polymorphic intersexed flat-abdomen wasps according to claim 4, characterized in that: The method of using the adult of the flat-bellied wasp carrying Rickettsia as a parent specifically includes: Adult flat-bellied wasps carrying Rickettsia are used as parents, and the emergence time of the parents is less than 6 hours.

6. The method for obtaining polymorphic intersexed flat-abdomen wasps according to claim 4, characterized in that: The method of feeding the parent with a rifampicin solution for several days specifically comprises: Feeding the parent with a rifampicin solution having a rifampicin concentration of 0.1 to 10.0 mg / mL for several days; Wherein, the rifampicin solution comprises a fructose solution containing rifampicin.

7. The method for obtaining polymorphic intersexed flat-abdomen wasps according to claim 4, characterized in that: The step of feeding the rifampicin solution for 24±1h and giving the parent insect eggs for parasitism specifically comprises: After feeding the rifampicin solution for 24±1h, the parents are given tussah eggs for parasitism, and then fresh tussah eggs are replaced at intervals of 24±1h.

8. The method for obtaining polymorphic intersexed flat-abdomen wasps according to claim 4, characterized in that: The offspring show polymorphic intersex phenotypes on the 2nd, 3rd and 4th days of feeding the rifampicin solution, namely, the intersex flat-abdomen wasp is obtained, which specifically includes: On days 2, 3, and 4 of feeding the rifampicin solution, the offspring showed polymorphic intersex phenotypes; Individuals that have both female and male bee characteristics are classified as an intersexual, and the intersexuals are collected to obtain intersexual flat-bellied wasps.

9. A polymorphic intersex flat-bellied wasp, characterized in that: The polymorphic intersexual flat-abdomen wasp is constructed by a method for obtaining a polymorphic intersexual flat-abdomen wasp according to any one of claims 1 to 8.

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