Method for identifying ritagin from noctuid black egg bee hosts and breeding noctuid black egg bees on large scale

Abstract

The invention provides a method for identifying ritadine from a host of Noctuid black egg bees and breeding the Noctuid black egg bees on a large scale, and belongs to the technical field of biological prevention and control of agricultural pests. The invention provides a noctuid black egg bee host recognition ritadine, which is obtained by treating spodoptera frugiperda eggs with n-hexane and separating a liquid phase. According to the present invention, the Noctuid black-egg bee host recognition ritamine can domesticate Noctuid black-egg bees and improve the parasitism rate of corcyra cephalonica eggs and the eclosion rate of offspring bees, and the results of the embodiment show that after the Noctuid black-egg bees are domesticated, the parasitism rate of the F30 generation is 88.23%, the eclosion rate of the offspring bees is 89.92%, and the large-scale breeding of the Noctuid black-egg bees can be achieved.

Description

Technical Field

[0001] This invention belongs to the field of biological control technology for agricultural pests, specifically relating to a method for host identification of luteolin by the black-egg wasp *Pteris vittata* and for large-scale breeding of *Pteris vittata*. Background Technology

[0002] Noctuidae, a family of Lepidoptera, are a major group of agricultural pests, including the beet armyworm (Spodoptera litura), fall armyworm (Spodopterafrugiperda), beet armyworm (Spodoptera exigua), armyworm (Mythimna separata), tobacco budworm (Helicoverpa assulta), and cotton bollworm (Helicoverpa armigera). Noctuidae pests have a wide distribution and host range, feeding on important crops such as corn, cotton, soybeans, wheat, and vegetables. They are characterized by high annual reproductive capacity, strong concealment, and the ability to cause severe outbreaks, resulting in significant damage to agricultural crops. The large number of egg masses laid by female noctuidae pests is the source of their damage in the field; therefore, the egg stage is a crucial window for noctuidae pest control. Utilizing highly efficient parasitic natural enemies during the egg stage is particularly important for the long-term control of these pests.

[0003] Telenomus remus, belonging to the family Scelionidae in the order Hymenoptera, is an important parasitic natural enemy of the egg stage of various Lepidoptera noctuid moths. Currently, some successful research experience has been achieved in the large-scale breeding of Telenomus remus using the eggs of natural hosts such as the beet armyworm and the cotton bollworm. However, the eggs of natural hosts are not durable and exhibit severe cannibalism in the larvae, resulting in excessively high costs for large-scale breeding. Therefore, a better alternative host is urgently needed.

[0004] The rice moth (Corcyra cephalonica), belonging to the family Galleriidae in the order Lepidoptera, is an ideal alternative host for the propagation of the black-egg wasp *Corcyra cephalonica* due to its ease of rearing, wide availability of feed, low cost, and suitability for large-scale production. However, rice moth eggs are not a natural host for *Corcyra cephalonica*, exhibiting low parasitism rates and low emergence rates of offspring wasps, severely hindering the large-scale breeding of *Corcyra cephalonica*. Summary of the Invention

[0005] In view of this, the present invention provides a host recognition alkaloid for the black-egg wasp of the noctuid moth, which can domesticate the black-egg wasp of the noctuid moth to increase the parasitism rate on rice moth eggs and the emergence rate of offspring wasps, thereby realizing the large-scale breeding of the black-egg wasp of the noctuid moth on the non-natural host rice moth eggs.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] This invention provides a host recognition alkaloid for the fall armyworm black egg wasp, which is obtained by treating fall armyworm eggs with n-hexane and then separating the liquid phase.

[0008] Preferably, the treatment comprises adding 0.4 to 2 mL of n-hexane to every 100 eggs of the fall armyworm.

[0009] Preferably, the fall armyworm eggs are those laid by the fall armyworm on the same day.

[0010] This invention provides an application of the host recognition lutein of the noctuid moth black egg wasp in the domestication of the noctuid moth black egg wasp.

[0011] Preferably, the domestication of the black-egg wasp includes promoting the parasitism of the black-egg wasp in rice moth eggs and / or promoting the emergence of the black-egg wasp offspring.

[0012] This invention provides a method for large-scale breeding of black egg wasps for moths, comprising the following steps:

[0013] The Noctuid moth black egg wasp was used as a host to identify rice moth eggs pretreated with luteolin, and then introduced female Noctuid moth black egg wasps for successive breeding.

[0014] Preferably, the pretreatment includes adding 5–20 μL of host-recognizing luteolin to every 100 rice moth eggs.

[0015] Preferably, the female black-egg wasp of the noctuid moth is a female black-egg wasp that has emerged within 24 hours.

[0016] Preferably, during the subsequent breeding, the ratio of female black-egg wasps of the noctuid moth to pretreated rice moth eggs is 1:(9-13).

[0017] Preferably, the subculture includes parasitic culture and artificial culture;

[0018] The parasitic culture time was 42-54 hours, the temperature was 25-27℃, the relative humidity was 60%-70%, the photoperiod light-dark ratio was 14:10, and the light intensity was 280-320 Lux;

[0019] The artificial culture temperature is 25–27°C, the relative humidity is 60%–70%, the photoperiod light-dark ratio is 14:10, and the light intensity is 280–320 Lux.

[0020] Compared with the prior art, the present invention has the following advantages:

[0021] This invention provides a host-recognition alkaloid for the fall armyworm (Spodoptera litura), obtained by treating fall armyworm eggs with n-hexane followed by liquid phase separation. This alkaloid can domesticate the fall armyworm, increasing its parasitism rate on Spodoptera litura eggs and improving the emergence rate of its offspring. In one embodiment, the residence time of the fall armyworm on Spodoptera litura eggs and the parasitism rate of the F1 generation were compared in different treatment groups. The results showed that, compared to the blank control group, the Spodoptera litura group, and the beet armyworm group, the treatment group treated with the alkaloid ... ， The emergence rate of the offspring wasps was 89.92%. Therefore, the host recognition alkaloid of the noctuid moth black egg wasp described in this invention can domesticate the noctuid moth black egg wasp, improve the parasitism rate on rice moth eggs and the emergence rate of offspring wasps, and realize the large-scale breeding of noctuid moth black egg wasps on non-natural host rice moth eggs.

[0022] This invention provides a method for the large-scale breeding of the black-egg wasp *Noctus muscarinatus*. The method uses rice moth eggs pretreated with a host-recognizing luteolin as the host, and then introduces female *Noctus muscarinatus* wasps for successive generations of breeding. This invention successfully induces *Noctus muscarinatus* to parasitize non-host rice moth eggs by improving the rice moth egg domestication method, resulting in the emergence of the next generation. Combined with continuous successive generations of *Noctus muscarinatus* parasitizing rice moth eggs pretreated with a host-recognizing luteolin, this significantly improves the parasitism rate and emergence rate of *Noctus muscarinatus* on rice moth eggs, enabling the large-scale breeding of *Noctus muscarinatus* using rice moth eggs as an alternative host. Furthermore, the method of this invention uses rice moth eggs as the propagation host for the black-egg wasp of the noctuid moth. The production technology of rice moth eggs is relatively mature and the production cost of rice moth eggs is low, which can realize the large-scale breeding of the black-egg wasp of the noctuid moth. The method of this invention utilizes rice moth eggs to breed the black-egg wasp of the noctuid moth on a large scale and releases it in the field, which can control the occurrence of noctuid moth populations, reduce its damage to crops, reduce the amount of chemical pesticides used, and improve the yield and quality of agricultural products. Attached Figure Description

[0023] Figure 1 The graph shows the retention time of the black egg wasp on the surface of the rice moth egg. * indicates a significant difference compared with the control group (P<0.05), and ** indicates an extremely significant difference compared with the control group (P<0.01).

[0024] Figure 2 The results of parasitism rate and emergence rate of rice moth eggs by the F1, F10 and F30 generations of black-egg wasp are shown in the figure. * indicates a significant difference compared with the fall armyworm group (P<0.05). Detailed Implementation

[0025] This invention provides a host recognition alkaloid for the fall armyworm black egg wasp, which is obtained by treating fall armyworm eggs with n-hexane and then separating the liquid phase.

[0026] In this invention, the fall armyworm eggs are preferably eggs laid by the fall armyworm on the same day, as eggs laid on the same day have the advantage of yielding a higher concentration of lutein. The treatment preferably includes adding 0.4–2 mL of n-hexane per 100 fall armyworm eggs, more preferably adding 0.6–1.5 mL of n-hexane, and most preferably adding 0.8 mL of n-hexane. After adding the n-hexane, it is preferable to gently shake and then let it stand; the shaking method preferably includes agitation, and the agitation time is preferably 1–5 minutes, which in specific embodiments can be 1 minute, 2 minutes, 3 minutes, 4 minutes, or 5 minutes; the standing time is preferably 8–12 minutes, which in specific embodiments can be 8 minutes, 9 minutes, 10 minutes, 11 minutes, or 12 minutes. The method for separating the liquid phase preferably includes drawing the clear liquid using a syringe; after separating the liquid phase, it is also preferred to concentrate the liquid phase, preferably by concentration with high-purity nitrogen (purity of 99.999%); the volume ratio of the concentrated liquid to the unconcentrated liquid is preferably (0.25-0.75):1, more preferably (0.4-0.6):1, and most preferably 0.5:1. The host recognition lutein of the noctuid moth black egg wasp is preferably stored at -20°C.

[0027] The host-recognition methylphenidate of the noctuid moth provided by this invention can domesticate the noctuid moth, increasing its parasitism rate on rice moth eggs and the emergence rate of its offspring. In one embodiment of this invention, the residence time of the noctuid moth on rice moth eggs and the parasitism rate of the F1 generation were compared in different treatment groups. The results showed that, compared with the blank control group, the beet armyworm group, and the sugar beet armyworm group, the noctuid moth host-recognition methylphenidate treatment group of this invention had the longest residence time and the highest parasitism rate. In another embodiment of this invention, the parasitism rate and emergence rate of the noctuid moth on rice moth eggs in the F1, F10, and F30 generations of the noctuid moth were compared in different treatment groups. The results showed that, compared with the control group, the beet armyworm group, and the sugar beet armyworm group, the noctuid moth host-recognition methylphenidate treatment group of this invention had the highest parasitism rate and emergence rate, with an F30 generation parasitism rate of 88.23%. ， The emergence rate of the offspring wasps was 89.92%. Therefore, the host recognition alkaloid of the noctuid moth black egg wasp described in this invention can domesticate the noctuid moth black egg wasp, improve the parasitism rate on rice moth eggs and the emergence rate of offspring wasps, and realize the large-scale breeding of noctuid moth black egg wasps on non-natural host rice moth eggs.

[0028] Based on the fact that the host recognition alkaloid of the noctuid moth black egg wasp has the function of domesticating the noctuid moth black egg wasp and improving the parasitism rate of rice moth eggs and the emergence rate of offspring wasps, the present invention provides an application of the host recognition alkaloid of the noctuid moth black egg wasp in the domestication of the noctuid moth black egg wasp.

[0029] In this invention, the domestication of the black-egg wasp of the noctuid moth preferably includes promoting the parasitism of the black-egg wasp in the eggs of the rice moth and / or promoting the emergence of the offspring of the black-egg wasp of the noctuid moth.

[0030] This invention provides a method for large-scale breeding of black egg wasps for moths, comprising the following steps:

[0031] The Noctuid moth black egg wasp was used as a host to identify rice moth eggs pretreated with luteolin, and then introduced female Noctuid moth black egg wasps for successive breeding.

[0032] In this invention, the pretreatment preferably includes adding 5-20 μL of a host-recognizing phytotoxicant to every 100 rice moth eggs, more preferably adding 8-15 μL, and can be up to 10 μL. The rice moth eggs are preferably eggs laid by the rice moth on the same day, as eggs laid on the same day are beneficial for improving the parasitism effect of the phytotoxicant. The rice moth eggs are preferably fixed on a flat surface, more preferably fixed on a cardboard card coated with glue, which facilitates the removal of the rice moth eggs.

[0033] In some specific embodiments, the feed for rice moth larvae preferably includes the following components: corn flour, wheat bran, soybean flour, yeast powder, white sugar, and compound vitamins;

[0034] The multivitamin includes vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin C, vitamin D, vitamin E, calcium pantothenate, and niacinamide.

[0035] In some specific embodiments, the corn flour preferably accounts for 60% to 90% of the feed mass, more preferably 65% ​​to 90%, 70% to 90%, 70% to 85%, 75% to 85%, 78% to 82%, and more specifically 78%, 79%, 80%, 81%, or 82%.

[0036] The wheat bran preferably accounts for 4-11% of the feed weight. In some specific embodiments, the weight percentage of wheat bran can be 4%-10%, 5%-10%, and more specifically 5%, 6%, 7%, 8%, 9%, or 10%.

[0037] The soybean meal preferably accounts for 4% to 20% of the feed weight. In some specific embodiments, the weight percentage of soybean meal can be 4% to 18%, 6% to 16%, 8% to 12%, and more specifically, 8%, 9%, 10%, 11%, or 12%.

[0038] The yeast powder preferably accounts for 1% to 5% of the feed weight. In some specific embodiments, the weight percentage of yeast powder can be 1% to 5%, and more specifically, it can be 1%, 2%, 3%, 4% or 5%.

[0039] The white sugar is preferably 0.5% to 2% of the feed weight, and in some specific embodiments, the weight percentage of white sugar can be 1% to 1.5%.

[0040] The compound vitamins preferably account for 0.5% to 2% of the feed weight. In some specific embodiments, the weight percentage of the compound vitamins can be 0.5% to 1.8%, 0.7% to 1.5%, 0.8% to 1.2%, and more specifically, 0.8%, 0.9%, 1%, 1.1%, or 1.2%.

[0041] The preferred mass ratio of vitamin A: vitamin B1: vitamin B2: vitamin B6: vitamin C: vitamin D: vitamin E: calcium pantothenate: nicotinamide in the compound vitamin is (0.1-2):(1-5):(0.1-2.5):(0.1-1):(15-40):(3-10):(0.1-2.5):(0.1-2.5):(0.1-2.5), and may also be (0.3-2):(1.5-4):(0.3-2):(0.2-0.8):(20-45):(5-8):(0.3-2):(0.3-2):(0.3-2).

[0042] In some specific embodiments, the method for preparing the feed includes the following steps:

[0043] (1) Place corn flour, wheat bran, soybean flour, yeast powder and white sugar in a high-pressure sterilizer at a temperature of 80-120℃ and a pressure of 100-110kPa for 60-90 minutes to sterilize;

[0044] (2) After each component has cooled, place it in a tray and mix it according to the ratio between each component. Finally, add compound vitamins and mix evenly to obtain the feed.

[0045] In this invention, the female black-egg wasp of the noctuid moth is preferably a female black-egg wasp of the noctuid moth that emerges within 24 hours, and more preferably a female black-egg wasp of the noctuid moth that has emerged within 24 hours after mating.

[0046] In this invention, during the subgeneration and breeding process, the preferred ratio of female black-egg wasps (*Hypericum noctuidum*) to pretreated rice moth eggs is 1:(9-13). Within this ratio range, it is beneficial for the black-egg wasps to fully parasitize the rice moth eggs. In specific embodiments, the ratio of wasp to eggs can be 1:10, 1:11, or 1:12.

[0047] In this invention, the subculture preferably includes parasitic culture and artificial culture. The parasitic culture time is preferably 42-54 hours, but can be 46 hours, 48 ​​hours, or 50 hours. The parasitic culture temperature is preferably 25-27°C, but can be 26°C. The relative humidity of the parasitic culture is preferably 60%-70%, but can be 65%. The photoperiod-to-dark ratio is preferably L:D = 14:10, and the light intensity is preferably 280-320 Lux, but can be 300 Lux. The temperature, relative humidity, photoperiod-to-dark ratio, and light intensity of the artificial culture are preferably the same as those of the parasitic culture. Under the specific parasitic and artificial culture conditions of this invention, it is beneficial to improve the parasitism rate and promote the emergence of black egg wasps from the noctuid moth. The artificial culture equipment preferably includes equipment that can control temperature, humidity, and light, such as an artificial climate chamber.

[0048] In one embodiment of the present invention, the parasitism rate and emergence rate of *Spodoptera litura* eggs by different treatment groups F1, F10, and F30 were compared. The results showed that, compared with the control group, the *Spodoptera litura* group, and the *Spodoptera litura* group, the parasitism rate and emergence rate of *Spodoptera litura* eggs in the treatment group described in the present invention were the highest, with the parasitism rate of *Spodoptera litura* eggs in the F30 generation reaching 88.23%. ， The emergence rate of the offspring wasps was 89.92%. This demonstrates that the method described in this invention can domesticate the black-egg wasp of the noctuid moth, increasing its parasitism rate on rice moth eggs and the emergence rate of its offspring wasps, thus enabling large-scale breeding of the black-egg wasp on the non-natural host rice moth eggs.

[0049] To further illustrate the present invention, the following detailed description, in conjunction with the accompanying drawings and embodiments, describes a method for host identification of luteolin and large-scale breeding of *Noctus molluscella*, provided by the present invention. However, these descriptions should not be construed as limiting the scope of protection of the present invention.

[0050] Unless otherwise specified, the techniques or conditions described in the following embodiments are generally performed in accordance with conventional techniques or conditions described in the literature in this field, or in accordance with the product manual and the manufacturer's recommendations.

[0051] Unless otherwise specified, all starting materials, ingredients and reagents are either commercially available or synthesized according to known methods.

[0052] Example 1

[0053] Establish a rice moth strain population of the black egg wasp *Noctus muscarinatus*.

[0054] 1. Preparation of host recognition lutein from crude extracts of eggs of different Noctuidae insects by the black egg wasp of Noctuidae.

[0055] Collect 500 eggs each from the fall armyworm, beet armyworm, and cabbage looper on the same day. Use a brush to sweep the scales of the eggs into a vial, then add 4 mL of n-hexane, seal the vial, gently shake for 1 minute, and let stand for 10 minutes. Afterward, use a syringe to aspirate the supernatant and concentrate it to 2 mL under high-purity nitrogen. The obtained host-recognizing luteolins from different noctuid moths are stored at -20°C for later use.

[0056] 2. Obtaining Rice Moth Eggs

[0057] The feed for rice moth larvae consists of the following components: corn flour, wheat bran, soybean flour, yeast powder, white sugar, and compound vitamins;

[0058] Corn flour, wheat bran, soybean flour, yeast powder, white sugar, and compound vitamins accounted for 78%, 7%, 12%, 1%, 1%, and 1% of the total feed weight, respectively.

[0059] A multivitamin consists of vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin C, vitamin D, vitamin E, calcium pantothenate, and niacinamide.

[0060] The adult rice moth is fed with 15% honey water.

[0061] The method for raising adult rice moths is as follows: After the rice moths emerge, they are placed in oviposition cages. Each cage contains 20 male and 20 female adults. The rearing conditions are set at a temperature of (26±2)℃, humidity of 70%±10%, and a day / night light duration of 14L:10D. The honey water is changed every 3 days. The oviposition cage is a cylindrical cage 60cm long and 15cm in diameter, with round wooden boards at both ends. The upper cage opening has a movable cover. The cage body is made of 40-mesh metal mesh, which is fixed to the circumference of the upper and lower cage boards.

[0062] 3. Effect of luteolin on host recognition of the black-egg wasp *Noctus muscarinatus* and its residence time.

[0063] In the treatment group, 50 rice moth eggs produced on the same day were collected and fixed on a 2cm side cardboard coated with glue. 10μL of host recognition lutein from different noctuid moths was added to the rice moth eggs and were respectively named the fall armyworm group, the beet armyworm group, and the sugar beet armyworm group.

[0064] Each group of egg cards was placed into different finger-shaped tubes. One female *Noctuidae* wasp that had emerged within 24 hours was placed in each tube, and the tube was then sealed. Timing was started simultaneously to record the time the female wasp remained on the egg mass. If the female wasp left the filter paper within 5 minutes and did not return within 20 minutes, the observation for that replicate was stopped; if it returned within 20 minutes, the observation continued. Rice moth eggs served as the control group. After every 5 wasps tested, the positions of the treatment group and the control group were exchanged to avoid the influence of position or other factors on the results. The experiment was repeated 20 times. The effect of host recognition lutein from different *Noctuidae* insects on the retention time of *Noctuidae* wasps on rice moth eggs was statistically analyzed. The results are shown below. Figure 1 As shown.

[0065] according to Figure 1 The results showed that the host recognition lutein of the black-egg wasp from different sources had a significant impact on the retention time of the black-egg wasp. Among them, the fall armyworm egg group and the beet armyworm group showed extremely significant differences from the control (P<0.01). The fall armyworm group had the longest retention time, at 223s, which was 170s longer than the control group's 53s. The beet armyworm group had a retention time of 178s, which was 125s longer than the control group. The beet armyworm group had a retention time of 72s, which was 19s longer than the control group.

[0066] 4. Parasitism of rice moth eggs by the black-egg wasp *Triplophysa maxima*.

[0067] In the treatment group, 50 rice moth eggs produced on the same day were collected and fixed on a 2cm side cardboard coated with glue. 10μL of host recognition lutein from different noctuid moths was added to the rice moth eggs and were respectively named the fall armyworm group, the beet armyworm group, and the sugar beet armyworm group.

[0068] Egg cards from each group were placed into different finger-shaped tubes, and female *Noctus spp.* that had emerged within 24 hours were introduced at a wasp-egg ratio of 1:10, while providing them with nutrition in a 15% honey water solution. After 48 hours of parasitism, the rice moth eggs were removed and placed in an artificial climate chamber with environmental conditions of 27℃, RH=65%, L:D=14:10h, and light intensity of 300 Lux. Hatching larvae were promptly removed. The egg masses continued to be cultured until the eggs turned black, indicating successful parasitism. After 8 days of parasitism, the parasitism was observed under a stereomicroscope, and the number of parasitized eggs was counted. Rice moth eggs were used as a control. The parasitism rate and emergence rate of *Noctus spp.* eggs were calculated using Formula I and Formula II, and the statistical results are shown in Table 1.

[0069] Parasitism rate (%) = Number of parasitic eggs / Total number of eggs × 100% Formula I

[0070] Emergence rate (%) = Number of emerged bees / Number of parasitic eggs × 100% (Formula II)

[0071] Table 1. Parasitism of rice moth eggs by the black egg wasp of the noctuid moth.

[0072] deal with Parasitism rate (%) Feathering rate (%) control group 0 0 Fall armyworm group 14.26±1.38a 23.42±2.50a Group Spodoptera litura 10.78±0.97b 19.65±1.38b Beet armyworm egg group 5.62±0.56c 7.17±0.53c

[0073] Note: Different letters indicate significant differences between different treatments (P<0.05).

[0074] In the treatment group using the host recognition luteolin of the fall armyworm wasp, the wasp was able to parasitize a small number of rice moth eggs, while the rice moth eggs in the control group showed no parasitism even after 7 parasitism attempts. The parasitism rate and emergence rate of rice moth eggs in the fall armyworm group were significantly higher than those in the beet armyworm and sugar beet armyworm groups. Subsequently, the emerged adult wasps were used as parent wasps for further parasitism, and the first batch of fall armyworm wasps bred was the F1 generation.

[0075] Example 2

[0076] Parasitism and emergence of rice moth eggs treated with luteolin by black egg wasps of different domestication generations of noctuid moths.

[0077] Fifty rice moth eggs produced on the same day were collected and fixed onto a 2cm side cardboard coated with glue. 10μL of host recognition lutein from different Noctuidae insects prepared in Example 1 was added to the rice moth eggs and labeled as the fall armyworm group, the beet armyworm group, and the sugar beet armyworm group, respectively.

[0078] Each group of egg cards was placed into different finger-shaped tubes. The *Noctus mollusc* wasps that emerged after parasitizing the rice moth eggs were designated as the F1 generation. F1 generation female *Noctus mollusc* wasps emerging within 24 hours were introduced at a wasp-to-egg ratio of 1:10, while simultaneously providing nutrition with 15% honey water. After 48 hours of parasitism, the rice moth eggs were removed and placed in an artificial climate chamber with environmental conditions of 27℃, RH=65%, L:D=14h:10h, and light intensity of 300 Lux. Hatching larvae were promptly removed. The egg masses continued to be cultured until the eggs turned black, indicating successful parasitism. After 8 days of parasitism, the parasitism was observed under a stereomicroscope, and the number of parasitized eggs was counted. Adult wasps emerged, and the emergence process was observed under a stereomicroscope, and the number of emerging adult wasps was counted. The above steps were repeated, using rice moth eggs treated with host-identifying luteolin as the host to domesticate the *Noctus mollusc* wasps to the F10 and F30 generations. The parasitism rate and emergence rate of the black-egg wasp on the rice moth eggs were calculated using Formula I and Formula II.

[0079] The parasitism and emergence of rice moth eggs by the F1, F10, and F30 generations of the fall armyworm, beet armyworm, and sugar beet armyworm groups are as follows: Figure 2 As shown.

[0080] Depend on Figure 2The results showed that the parasitism rates of the fall armyworm (F1, F10, and F30 generations) on rice moth eggs were the highest, at 14.26%, 23.57%, and 88.23%, respectively. Figure 2 In the F1, F10, and F30 generations of the black-egg wasp *Noctus cuspidatus*, the parasitism rate increased significantly with increasing domestication generations; the emergence rates of the F1, F10, and F30 generations were 23.42%, 28.68%, and 89.92%, respectively. Figure 2 In the case of B), the molting rate also increases significantly with the increase of the number of domestication generations.

[0081] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. Other embodiments can be obtained based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A host recognition phenanthrene of the black-egg wasp *Pterocarya stenoptera*, characterized in that, It is obtained by treating fall armyworm eggs with n-hexane and then separating the liquid phase.

2. The host recognition phenotype of the black-egg wasp according to claim 1, characterized in that, The treatment involves adding 0.4–2 mL of n-hexane to every 100 of the fall armyworm eggs.

3. The host recognition lutein of the black-egg wasp according to claim 1 or 2, characterized in that, The eggs mentioned are those laid by the fall armyworm on that day.

4. The application of the host recognition lutein of the black-egg wasp according to any one of claims 1 to 3 in the domestication of the black-egg wasp.

5. The application according to claim 4, characterized in that, The domestication of the black-egg wasp includes promoting the parasitism of the black-egg wasp in rice moth eggs and / or promoting the emergence of the black-egg wasp offspring.

6. A method for large-scale breeding of black egg wasps for the noctuid moth, characterized in that, Includes the following steps: Using the rice moth eggs pretreated with luteolin as the host identification agent described in any one of claims 1 to 3, female *Noctus black ovipositor* wasps were introduced for subgeneration.

7. The method according to claim 6, characterized in that, The pretreatment includes adding 5–20 μL of host-recognizing luteolin to every 100 rice moth eggs.

8. The method according to claim 6, characterized in that, The female Noctuid moth egg wasp mentioned refers to the female Noctuid moth egg wasp that emerged within 24 hours.

9. The method according to claim 6, characterized in that, During the subsequent breeding, the ratio of female black-egg wasps of the noctuid moth to pretreated rice moth eggs was 1:(9-13).

10. The method according to any one of claims 6 to 9, characterized in that, The subsequent breeding includes parasitic culture and artificial culture; The parasitic culture time was 42-54 hours, the temperature was 25-27℃, the relative humidity was 60%-70%, the photoperiod light-dark ratio was 14:10, and the light intensity was 280-320 Lux; The artificial culture temperature is 25–27°C, the relative humidity is 60%–70%, the photoperiod light-dark ratio is 14:10, and the light intensity is 280–320 Lux.

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