Method for environmentally-friendly control of ectropis grisescens warren

Abstract

The present invention belongs to the field of crop plant protection. Disclosed is a method for environmentally-friendly control of Ectropis grisescens Warren. The present invention aims to suppress field populations of wild Ectropis grisescens Warren by means of obtaining sterile Ectropis grisescens Warren adults using X-ray irradiation, thereby reducing the use of chemical pesticides and protecting the ecological environment of tea plantations. The present invention determines the physiological effects of different irradiation doses on Ectropis grisescens Warren pupae and adults, establishes the relationship between the sterility rate and irradiation dose, and screens the optimal irradiation dose for obtaining sterile Ectropis grisescens Warren. In addition, further provided in the present invention is a method for delaying the emergence of Ectropis grisescens Warren pupae at a low temperature, thereby realizing the synchronized emergence of sterile insects, enhancing the operability of the method, and optimizing the effect of field release. Field release experiments show that irradiation-treated sterile Ectropis grisescens Warren can significantly inhibit the insect count, which verifies the good control effect of the method. Moreover, the quality of fresh leaves from tea plantations with the release of the sterile insects is significantly improved.

Description

A green method for controlling the tea geometrid moth Technical Field

[0001] This invention relates to a green method for controlling the gray tea geometrid moth, belonging to the field of crop plant protection. Background Technology

[0002] The grey tea looper (Ectropis grisescens Warren), belonging to the family Geometridae in the order Lepidoptera, is a major leaf-eating pest in tea gardens. Characterized by high generation rates, rapid reproduction, and fast spread, the grey tea looper is highly susceptible to outbreaks, posing a significant threat to tea gardens. Currently, chemical pesticides remain the primary means of controlling the grey tea looper in tea gardens. However, the overuse of chemical pesticides pollutes the ecological environment of tea gardens and kills natural enemies of tea pests, severely hindering the development of the tea industry.

[0003] Insect sterilization is an environmentally friendly strategy for controlling pests and plays an irreplaceable role in integrated pest management. This technology involves continuously releasing treated sterile insects into the natural environment to mate with wild insects in the field. The resulting eggs either do not hatch or do not hatch completely, thus suppressing the target insect population and controlling it within a threshold that does not impact human production and the economy. Sterile insects are typically produced through irradiation; when released into the target area and mating with normal insect populations in the wild, their offspring also fail to develop normally or reproduce successfully.

[0004] In existing techniques for inducing insect sterility through irradiation, male and female pupae need to be separated before irradiation. However, researchers, for ecological safety reasons, only release irradiated male pupae into the wild to suppress wild insect populations.

[0005] For example, in a method for controlling the sterility of spotted-winged fruit flies by irradiation, as described in publication number CN109105341A, the method involves feeding and allowing spotted-winged fruit flies to lay eggs to obtain 4-day-old pupae; irradiating the 4-day-old pupae; after irradiation, the pupae emerge, and male spotted-winged fruit flies are selected; and the selected male spotted-winged fruit flies are released into the wild.

[0006] For example, a method for biological control of the invasive coconut weaver moth, as described in publication number CN109729893A, involves irradiating the early pupae of male coconut weaver moths with a cobalt-60 irradiation source at a dose of 100–200 Gy for 1–2 hours; placing the pupae at a temperature of 25–27°C and a relative humidity of 70% ± 10% until they emerge as adults; then transferring the adults into glass tubes and rearing them in irradiated coconut leaves to obtain sterile males; and finally releasing the sterile males based on forest monitoring of the number of female coconut weaver moth adults.

[0007] For example, in a method for X-ray irradiation sterilization for controlling bollworms, published in CN114467861A, the method involves rearing bollworms indoors using artificial feed to obtain bollworm pupae aged 1-7 days; irradiating the bollworm pupae at an irradiation dose of 134-168 Gy; obtaining irradiated pupae; placing the irradiated pupae in a cage; and after the pupae emerge, selecting male bollworm adults with normal appearance and activity; and releasing the selected male bollworm adults into their natural population.

[0008] For example, CN114946482A describes a radiation-induced sterilization method for controlling the rice stem borer. The method involves raising rice stem borers to obtain pupae; grouping the pupae by instar and setting a rearing temperature; selecting healthy male pupae 48 hours before emergence and continuing to raise them; pre-treating the healthy male pupae before irradiation; placing the selected healthy male pupae in an irradiation container in an irradiation chamber for irradiation sterilization; after irradiation, the rice stem borers are returned to a normal rearing environment to emerge, and the adults are released into the wild.

[0009] For example, in a method for obtaining sterile male tomato leafminer moths described in publication number CN115088681A, the method involves using... 137 Tomato leafminer male pupae were treated with Cs-γ rays to obtain sterile male adults of the tomato leafminer; the irradiation dose was 200-300 Gy; the irradiation dose rate was 0.8-1.2 Gy / min.

[0010] The method of manually selecting male adults and releasing them into the wild not only requires professionals to distinguish between female and male pupae, but also increases costs and operational difficulty. Furthermore, the pupal stage plays a crucial role in insect sterilization techniques. Insects at different pupal stages emerge at different times, and releasing sterile insects into the wild requires relatively concentrated emergence to achieve better control effects. However, due to the inconsistency in insect growth, collecting pupae of the same pupal stage is difficult, severely affecting irradiation efficiency and the acquisition of sterile insects. Because different insects have varying tolerances to X-rays, the optimal irradiation dose for sterilization differs for each insect species. Therefore, selecting the optimal sterilization dose is a critical step in implementing insect sterilization techniques. To date, there are no reports, domestic or international, on the use of X-ray irradiation for sterilization of the tea geometrid moth, nor on the evaluation of its field control effects and production benefits. Summary of the Invention

[0011] The purpose of this invention is to overcome the shortcomings of the prior art and provide an X-ray irradiation sterilization method for controlling the tea looper. The X-ray irradiation source based on this invention has advantages over gamma-ray irradiation sources, such as being cleaner and more environmentally friendly, simpler to operate, more efficient, and without safety hazards, and has broad application prospects. However, there are currently no reports on the application value of X-ray irradiation sources for the tea looper.

[0012] This invention first designs a method for obtaining sterile adult gray tea geometrid moths using X-rays, comprising the following steps:

[0013] (1) Collect the pupae of the gray tea geometrid moth and distinguish between males and females;

[0014] (2) Select late-stage pupae and irradiate them under different irradiation dose gradients;

[0015] (3) Place the irradiated pupae into a 1000mL rearing box containing moist fine sand and observe their emergence.

[0016] (4) Record the number of dead pupae, the number of normal emergence pupae, and the number of deformed pupae; calculate the deformed pupae and the corrected mortality rate.

[0017] (5) Record the physiological indicators of the adult moths that emerged from the irradiated pupae. The adult moths that emerged from the irradiated pupae were paired with adult moths of the opposite sex that were not irradiated. The mating rate, number of eggs laid and number of eggs hatched were recorded to obtain the sterility rate. The sterility rate of the irradiated moths was fitted by the Logistic regression model.

[0018] (6) Based on the physiological indicators and sterility rate fitting equation of the irradiated tea geometrid moth, the optimal sterility dose of the tea geometrid moth was obtained, and sterile adult tea geometrid moths were obtained using the optimal sterility dose.

[0019] (7) The growth and reproduction of offspring under the optimal sterility dose of the gray tea geometrid moth were analyzed by age-stage sex life tables.

[0020] To achieve a relatively concentrated emergence of sterile insects released into the wild and increase the ease of pupa acquisition, this invention provides a method for delaying the emergence time of insect pupae, based on the insects' overwintering habits and low-temperature diapause. The method includes the following steps:

[0021] (1) Select 5-day-old pupae of the gray tea geometrid moth and group them;

[0022] (2) Store in a refrigerator at 4°C for low temperature;

[0023] (3) After 48 hours of low-temperature treatment, the treatment group and the control group were irradiated with X-rays at the optimal infertility dose.

[0024] (4) Record the emergence rate and emergence time of each group of sterile pupae.

[0025] Based on the above research results, it was found that under the optimal sterility dose of 184.65 Gy, the sterility rate of female adult tea geometrid moths is >99%, and male tea geometrid moths can mate multiple times. Based on these characteristics, this invention provides a field release method, including the following steps:

[0026] (1) Raising and collecting male and female pupae of the gray tea geometrid moth;

[0027] (2) The male and female pupae of the gray tea geometrid moth were irradiated together at the optimal sterility dose of 184.65 Gy;

[0028] (3) Release the irradiated tea geometrid caterpillar pupae into the affected tea gardens.

[0029] To investigate the impact of this method on tea garden production efficiency, this invention provides a method for assessing the quality of damaged fresh leaves. It includes the following steps:

[0030] (1) Take ten sample plots each from tea gardens where the sterile tea geometrid moth was released and from normal tea gardens;

[0031] (2) Collect fresh leaves from the middle and upper parts of two tea garden sample plots;

[0032] (3) Weigh and record the data to analyze the damage to the fresh leaves.

[0033] This invention also provides a method for preventing and controlling the gray tea geometrid moth through X-ray irradiation sterilization, the method comprising the following steps:

[0034] (1) The gray tea geometrid moth was raised indoors using artificial feed breeding method to obtain gray tea geometrid moth pupae aged 5 to 7 days.

[0035] (2) The pupae of the tea geometrid moth were irradiated at an irradiation dose of 160–200 Gy to obtain irradiated pupae;

[0036] (3) The pupae obtained after irradiation do not need to be separated into pupae. They are released directly into their natural population to compete with wild male or female insects for mating with wild female or male insects, so that wild female insects do not lay eggs or the eggs they lay do not hatch or reduce the number of eggs laid.

[0037] In one embodiment of the present invention, in step (2), before irradiation, the pupae of the gray tea geometrid moth are collected, and the pupae at different time periods are subjected to low-temperature treatment to regulate the emergence period. The specific method is as follows: the pupae of the gray tea geometrid moth that are 5 to 7 days old are placed in a refrigerator at 4 to 8°C for low-temperature treatment for 0 to 72 hours and then subjected to irradiation treatment.

[0038] In one embodiment of the present invention, 5-7 day old gray tea geometrid moth pupae are placed in a 4°C refrigerator for low-temperature treatment for 48 hours and then subjected to irradiation treatment.

[0039] In one embodiment of the present invention, the irradiation procedure is as follows: the pupa is placed in a self-sealing bag for irradiation treatment, and a wad of moist cotton is placed inside to maintain humidity; the irradiation dose rate is 1.0 to 1.2 Gy / min.

[0040] In one embodiment of the present invention, the method for obtaining the gray tea geometrid moth pupa in step (1) is as follows:

[0041] 1) Collect wild larvae of the tea geometrid moth, feed them with fresh tea branches, and raise them indoors to expand the population.

[0042] 2) Collect egg masses. After the eggs hatch, transfer the newly hatched larvae of the tea geometrid moth in batches to a culture tray containing fresh tea branches. Culture them for 14 to 20 days under the conditions of an ambient temperature of 24±2℃, a humidity of 60 to 70%, and a photoperiod of 16L:8D until the larvae reach the mature stage.

[0043] 3) Mark the pupation date after the mature larvae pupate, and collect the pupae of the gray tea geometrid moth that are 5 to 7 days old;

[0044] In one embodiment of the present invention, the irradiated pupae are released into their natural population 3 to 5 times;

[0045] In one embodiment of the present invention, the irradiation dose is 184.65 Gy.

[0046] The present invention also provides a method for field release of the tea geometrid moth by irradiation, the method comprising the following steps:

[0047] (1) The gray tea geometrid moth was raised indoors using artificial feed breeding method to obtain gray tea geometrid moth pupae aged 5 to 7 days.

[0048] (2) The pupae of the tea geometrid moth were irradiated at an irradiation dose of 160–200 Gy to obtain irradiated pupae;

[0049] (3) Release the irradiated pupae in the field.

[0050] In one embodiment of the present invention, in step (2), before irradiation, the pupae of the gray tea geometrid moth are collected, and the pupae at different time periods are subjected to low-temperature treatment to regulate the emergence period. The specific method is as follows: the pupae of the gray tea geometrid moth that are 5 to 7 days old are placed in a refrigerator at 4 to 8°C for low-temperature treatment for 0 to 72 hours and then subjected to irradiation treatment.

[0051] In one embodiment of the present invention, 5-7 day old gray tea geometrid moth pupae are placed in a 4°C refrigerator for low-temperature treatment for 48 hours and then subjected to irradiation treatment.

[0052] In one embodiment of the present invention, the irradiation procedure is as follows: the pupa is placed in a self-sealing bag for irradiation treatment, and a wad of moist cotton is placed inside to maintain humidity; the irradiation dose rate is 1.0 to 1.2 Gy / min.

[0053] In one embodiment of the present invention, the method for obtaining the gray tea geometrid moth pupa in step (1) is as follows:

[0054] 1) Collect wild larvae of the tea geometrid moth, feed them with fresh tea branches, and raise them indoors to expand the population.

[0055] 2) Collect egg masses. After the eggs hatch, transfer the newly hatched larvae of the tea geometrid moth in batches to a culture tray containing fresh tea branches. Culture them for 0 to 72 days under the conditions of an ambient temperature of 24±2℃, a humidity of 60-70%, and a photoperiod of 16L:8D until the larvae reach maturity.

[0056] 3) Mark the pupation date after the mature larvae pupate, and collect the pupae of the gray tea geometrid moth that are 5 to 7 days old;

[0057] In one embodiment of the present invention, the irradiated pupae are released in the middle of a field, and the release is repeated 3 to 5 times.

[0058] In one embodiment of the present invention, the irradiation dose is 184.65 Gy.

[0059] In one embodiment of the present invention, the quadrat method is used to conduct a field insect population survey. Ten quadrats are set up in both the control group and the treatment group. Each quadrat is a 1×3 meter tea row and is numbered. The quadrat method and the pot tapping method are used to conduct the insect population survey. Three pots are tapped in each quadrat.

[0060] In one embodiment of the present invention, a five-point sampling method was used in the experimental fields of the control group and the treatment group to select five quadrats, each with an area of ​​0.5 × 0.5 meters, and collect all fresh leaves from the middle and above of the tea trees in the quadrats. Beneficial effects

[0061] 1. Compared to gamma rays, X-rays offer advantages such as being cleaner, more environmentally friendly, simpler to operate, and more efficient in the application of sterile insect technology. This invention provides an effective method for obtaining sterile insects using X-rays, demonstrating the feasibility of using X-rays to sterilize the tea geometrid moth. Furthermore, using X-ray irradiation for sterile control of the tea geometrid moth not only effectively reduces its reproductive capacity and population growth rate but also reduces the use of chemical pesticides, resulting in less negative environmental impact. This method provides strong support for establishing a green pest control system and contributes to the sustainable management of tea garden pests.

[0062] 2. Currently, in the field release phase of using insect sterility technology to control pests, the general method is to manually select male adults and then release them into the wild. Since releasing sterile insects in the field is a continuous and repeated process, requiring a large number of sterile insects to suppress the field population, the separation of pupae becomes a thorny problem. This not only requires professionals to distinguish between female and male pupae but also consumes a lot of time and effort, increasing costs and operational difficulty. This method utilizes the characteristic of the male and female pupae of the tea geometrid moth, which, after X-ray irradiation, simultaneously suppresses the density of the wild population. It innovatively adopts a method of releasing male and female pupae simultaneously after irradiation, greatly reducing costs and operational difficulty while improving the efficiency of population sterility.

[0063] This invention utilizes the low-temperature diapause behavior of the tea geometrid moth to propose a method for uniformly ecloding pupae through low-temperature treatment. Insects at different pupal stages eclode at different times. Due to the inconsistency in insect growth, collecting pupae at the same eclosion stage is difficult, severely impacting irradiation efficiency and the acquisition of sterile insects. This method uses low temperature to delay the eclosion time of early-stage pupae, achieving simultaneous eclosion with later-stage pupae. This effectively solves the problem of eclosion time differences caused by the inconsistency in insect growth, resulting in a more concentrated release and eclosion of sterile insects in the field, thus increasing the suppression effect on field pest populations.

[0064] 3. The application value of irradiation-based insect sterilization technology lies in its economic value, but the impact of this method on economic value has not yet been reported. In tea production and processing, the quality of fresh leaves affects tea yield, and tea yield is closely linked to tea farmers' income. This invention provides a method for measuring the economic value impact of insect sterilization technology, offering a reference for the economic benefits that this technology can bring. Attached Figure Description

[0065] Figure 1: Effects of different doses of X-ray irradiation on the pupae of the tea geometrid moth; where (A) represents the deformity rate and (B) represents the corrected mortality rate; data are mean ± standard deviation, and different lowercase letters on the columns indicate significant differences between the treatment groups and the control group at different irradiation doses.

[0066] Figure 2: Effects of different doses of X-ray irradiation on physiological indicators of the tea geometrid moth; where (A) is the eclosion time, (B) is the adult lifespan, (C) is the mating rate, and (D) is the number of eggs laid by a single female; data are mean ± standard deviation; N, no irradiation; R, irradiation treatment.

[0067] Figure 3: Logistic regression curves showing the relationship between crossbreeding sterility rate and X-ray irradiation dose in the gray tea geometrid moth; (A) shows the sterility rate of X-ray irradiated male adults paired with unirradiated male adults; (B) shows the sterility rate of X-ray irradiated female adults paired with unirradiated male adults; data are mean ± standard deviation, and different lowercase letters on the bars indicate significant differences in sterility rates among different irradiation dose treatment groups (p < 0.05, Tukey multiple comparisons).

[0068] Figure 4: Age characteristics, survival rate, and fertility of the F1 generation of irradiated male parents of the tea geometrid moth; (A) F1 generation of unirradiated parents; (BD) F1 generation of unirradiated female adults mated with male adults irradiated with X-rays at doses of 160 Gy, 184.65 Gy, and 200 Gy during the pupal stage; x f represents the survival rate for age-related characteristics. xj For age-stage characteristic reproductive capacity, m x For age-related reproductive capacity, l x m x Net reproductive rate for age characteristics.

[0069] Figure 5: Different dosages 60 Effects of Co-γ ray irradiation on the pupae of the tea geometrid moth; where (A) represents the deformity rate and (B) represents the corrected mortality rate; data are mean ± standard deviation, and different lowercase letters on the columns indicate significant differences between the treatment groups and the control group at different irradiation doses.

[0070] Figure 6: Different dosages 60 Effects of Co-γ ray irradiation on physiological indicators of the tea geometrid moth; where (A) is the eclosion time, (B) is the adult lifespan, (C) is the mating rate, and (D) is the number of eggs laid by a single female; data are mean ± standard deviation; N, no irradiation; R, irradiation treatment.

[0071] Figure 7: Cross-fertility rate of the brown geometrid moth and 60 The Co-γ ray irradiation dose-related Logistic fitting curve; (A) is... 60 The sterility rate of male adults irradiated with Co-γ rays paired with unirradiated male adults of the opposite sex; (B) is 60Sterility rate of female adult insects irradiated with Co-γ rays paired with unirradiated adult insects of the opposite sex; data are mean ± standard deviation, and different lowercase letters on the columns indicate significant differences in sterility rates among different irradiation dose groups (p < 0.05, Tukey multiple comparisons).

[0072] Figure 8: Dynamic changes in the population of larvae of the tea geometrid moth under different treatment conditions.

[0073] Figure 9: Dynamic changes in the population of larvae of the tea geometrid moth under different release methods. Detailed Implementation

[0074] The detection methods involved in the following embodiments:

[0075] malformation and corrected mortality

[0076] Adults that fail to unfold their wings or have incomplete wings after emergence are considered deformed. The deformed and corrected mortality rates of pupae are calculated based on the emergence status. Corrected mortality rate = treatment group mortality rate - control group mortality rate.

[0077] Infertility rate

[0078] Adult *Tea geometrid moth* pupae emerging from irradiated pupae at various dose gradients were paired with unirradiated adult *Tea geometrid moth* of the opposite sex and placed in interlocking plastic cups. Each cup contained one pair of adults and two fresh tea leaves. Eggs laid were counted and collected. Twenty pairs of adult *Tea geometrid moth* were paired for each treatment gradient. Dead female adults were dissected under a stereomicroscope to check for spermatophores, determining if mating had occurred. If no spermatophores were found during dissection but the eggs hatched normally, mating was considered effective, and the mating rate was calculated. The lifespan of each group of female and male adults was recorded. The number of eggs laid and unhatched were counted to determine the sterility rate. A logistic regression model was used to fit the sterility rate of *Tea geometrid moth* after irradiation.

[0079] Sub-infertility radiation dose

[0080] This usually refers to a state where organisms exhibit partial rather than complete sterility after irradiation treatment. High-dose irradiation causes sterility in insects, reducing their fitness and making them unable to compete for mates with the natural population, thus affecting pest control effectiveness. Therefore, in recent years, sub-sterile doses have been used to treat pests, which can improve the ability of irradiated insects to compete for mates. Through genetics, the irradiation-induced chromosomal translocation changes are passed on to the next generation, causing the next generation of pests to also lose their fertility, thus reducing the natural population.

[0081] sub-infertility rate

[0082] Generally, the higher the irradiation dose, the higher the sterility rate. To achieve a 100% sterility rate in male adult gray tea geometrid moths, a high irradiation dose is required. However, a high sterility dose will impair certain physiological indicators of the gray tea geometrid moths, which is not conducive to their release into the wild. Therefore, we use the irradiation dose that causes sub-sterility in gray tea geometrid moths as our optimal irradiation dose to balance the sterility rate and physiological indicators of the gray tea geometrid moths. This leads to the subsequent progeny culture to observe the evolution of the progeny population under the sub-sterile dose.

[0083] Sub-infertility is an infertility rate greater than 80%, while complete infertility is 100% infertility.

[0084] Age-related survival rate and age-stage fertility curves

[0085] The hatched F1 larvae were fed fresh tea branches, and their developmental status was observed daily to record the interphase of each stage. TWOSEX-MSChart software was used to statistically analyze the life table parameters of the F1 generation individuals. The mean and standard error of each parameter in the life table of the experimental population of *Tea geometrid moth* in the treatment and control groups were calculated using the paired bootstrap test (n=100000). The Pick 1 by 1 function in TWOSEX-MSChart software was used to compare the significant differences in age-stage characteristic survival rate and age-stage characteristic reproductive capacity between the two groups. Survival rate and reproductive rate curves were generated using OriginPro 2021.

[0086] Example 1: Screening of the optimal X-ray irradiation dose for the gray tea geometrid moth

[0087] The specific steps are as follows:

[0088] 1. Irradiation experiment

[0089] (1) Raising pupae:

[0090] Feed the larvae of the gray tea geometrid moth with fresh tea branches. Select 5-day-old larvae and place them in a rearing box. Record the pupation date after the larvae pupate.

[0091] (2) Irradiation:

[0092] On the second day after the larvae pupate, the sex of the pupae is distinguished under a stereomicroscope. Female and male pupae of the gray tea geometrid moth with a pupal stage of 5-6 days (late pupal stage) are selected and subjected to irradiation treatment respectively.

[0093] Six treatment groups (40, 80, 120, 160, 200 and 240 Gy) were designed for female and male pupae according to the irradiation dose, with 25 pupae in each group and two groups (male and female) in each treatment;

[0094] The irradiation procedure was as follows: the pupae were placed in a self-sealing bag for irradiation treatment, with a wad of moist cotton placed inside to maintain humidity; the pupae in the control group were not irradiated, and the irradiation dose rate was 1.0–1.2 Gy / min.

[0095] (3) Feathering:

[0096] The irradiated pupae were placed in 1000mL packaging boxes containing moist fine sand and their eclosion was observed.

[0097] 2. Experimental Results

[0098] The deformity and corrected mortality rates of pupae, adult physiological indicators, and sterility rates were statistically analyzed for each dose gradient. A logistic regression model was used to fit the sterility rate of *Tea geometrid moth* after irradiation. Based on the logistic regression analysis of the sterility rate after irradiation, the optimal sterility dose range for *Tea geometrid moth* was determined. The results are shown in Figures 1, 2, and 3, and Table 1.

[0099] Table 1: Summary of two-way ANOVA on physiological indicators of *Tea geometrid moth* after irradiation with different X-ray irradiation doses and sex.

[0100] The results show:

[0101] (1) Outer malformation rate: After X-ray irradiation of 200-240 Gy, the outer malformation rate of the pupae of the gray tea geometrid moth showed an increasing trend, while after X-ray irradiation of 40-160 Gy, the outer malformation rate of the pupae of the gray tea geometrid moth had no significant effect compared with the control group (Figure 1 (A)).

[0102] Corrected mortality rate: The corrected mortality rate of the pupa of the tea geometrid moth increased under X-ray irradiation of 200-240 Gy, while the corrected mortality rate of the pupa of the tea geometrid moth was not significantly affected by X-ray irradiation of 40-160 Gy compared with the control group (Figure 1, (B)).

[0103] (2) X-ray irradiation significantly affected the reproductive parameters and adult lifespan of the gray tea geometrid moth of different sexes.

[0104] Following X-ray irradiation, the eclosion time of the pupa of the gray tea geometrid moth varied with different irradiation doses (F... 6,266 =22.63, P<0.0001, there is a significant difference between (A) in Table 1 and Figure 2.

[0105] Under X-ray irradiation, the irradiation dose (F) 6,266 =2.724, P=0.0139, Table 1, Figure 2 (B)), sex (F) 1,266 =5.067, P=0.0252, Table 1, Figure 2 (B)), interaction between irradiation dose and sex (F 6,266=2.509, P=0.0223, Table 1 and Figure 2 (B) all significantly affected the adult lifespan of the gray tea geometrid moth.

[0106] Irradiation dose (F 6,266 =19.86, P<0.0001, Table 1, Figure 2 (D)), interaction between irradiation dose and sex (F 6,266 =5.228, P<0.0001, Table 1, Figure 2 (D)) significantly affected the egg production per female adult of the gray tea geometrid moth exposed to X-rays.

[0107] (3) X-ray irradiation dose (F) 6,266 =297.8, P<0.0001) and the sex of the gray tea geometrid moth (F 1,266 =89.73, P<0.0001) all significantly affected the sterility rate of the irradiated tea geometrid moth. After pairing irradiated adult tea geometrid moths with unirradiated adult tea geometrid moths of the opposite sex, the relationship between radiation dose and sterility rate could be well fitted using a logistic regression model (R0). 2 >0.95)(Figure 3(A); Figure 3(B)).

[0108] When normal female adult tea geometrid moths are paired with irradiated male adult tea geometrid moths, the irradiation dose range for obtaining a good F1 generation sub-sterility rate (>80%) after X-ray irradiation can be determined from the trend of the inflection point of the Logistic regression model curve to be 160-200 Gy (Figure 3(A)).

[0109] When normal male adult tea geometrid moths are paired with irradiated female adult tea geometrid moths, the irradiation dose range for obtaining a good F1 generation sub-sterility rate (>80%) after X-ray irradiation is 80-120 Gy, and the irradiation dose range for obtaining a completely sterile F1 generation (>99%) is 160-200 Gy (Figure 3(B)).

[0110] By fitting curves of sterility rates under various irradiation doses for male and female *Tea geometrid moth*, it was found that under the same irradiation dose, female *Tea geometrid moth* are more sensitive to X-rays, and their sterility rate is higher than that of males. Based on this finding and the fact that higher irradiation doses result in lower fitness of adult *Tea geometrid moth*, to ensure the ability of *Tea geometrid moth* to compete for mates with the natural population, we considered using the male sub-sterile dose as a benchmark to determine the optimal sterility dose range for *Tea geometrid moth*. Subsequently, by comprehensively analyzing the malformation rate and corrected mortality rate of irradiated pupae, as well as the emergence time, adult size, mating rate, sterility rate, and offspring population characteristics of adults after irradiation, the optimal irradiation dose was determined.

[0111] Example 2: Effects of X-ray irradiation on the F1 generation of male gray tea geometrid moths

[0112] According to Example 1, the optimal irradiation dose range for the tea geometrid moth is 160-200 Gy. The sterility rate of the tea geometrid moth was fitted using a Logistic regression model, yielding the fitted curve equation y = 0.872 / [1 + e^(1.839 - 0.023x)]. Based on this equation, the irradiation dose required for male adults to achieve 80% sterility was calculated to be 184.65 Gy. To determine the population changes in offspring after sub-sterile irradiation, we cultured and observed F1 individuals of the tea geometrid moth at three sub-sterile irradiation doses (160 Gy, 184.65 Gy, and 200 Gy) and collected age-stage sex life tables for systematic analysis, providing a reliable theoretical basis for the next stage of field release.

[0113] The specific steps are as follows:

[0114] 1. Irradiation experiment of male adult gray tea geometrid moth

[0115] (1) Raising pupae:

[0116] Feed the larvae of the gray tea geometrid moth with fresh tea branches. Select 5th instar larvae and place them in a rearing box. Record the pupation date after the larvae pupate.

[0117] (2) Irradiation:

[0118] On the second day after pupation, the sex of the pupae was distinguished under a stereomicroscope. Male pupae of the gray tea geometrid moth, with a pupal stage of 5-6 days (late pupal stage), were selected for irradiation treatment.

[0119] Male pupae were set up with irradiation doses of 160 Gy, 184.65 Gy, and 200 Gy, with 50 pupae in each group;

[0120] The irradiation procedure was as follows: the pupae were placed in a self-sealing bag for irradiation treatment, with a wad of moist cotton placed inside to maintain humidity; the pupae in the control group were not irradiated, and the irradiation dose rate was 1.0–1.2 Gy / min.

[0121] 2. Effects of irradiation on the F1 generation of the gray tea geometrid moth

[0122] (1) The irradiated male adult of the tea geometrid moth obtained in step 1 was mated with the unirradiated female adult of the tea geometrid moth.

[0123] Meanwhile, larvae obtained from the mating of male and female adult gray tea geometrid moths that have not been irradiated were used as a control (CK group).

[0124] (2) 100 hatched F1 larvae were fed fresh tea branches and their development was observed daily to record the interphase of each stage. Normal, unirradiated gray geometrid moth larvae were used as a control.

[0125] After the F1 generation pupae emerge, the emerging adults are paired with untreated male adults of the opposite sex, and the number of eggs laid and hatched in the F1 generation is recorded. The growth and reproduction of the F1 generation under the optimal sterile dose of irradiation for male gray tea geometrid moths are analyzed using life tables.

[0126] 3. Experimental Results

[0127] The results are shown in Tables 2, 3, and 4, and Figure 4:

[0128] Table 2: Effects of X-ray substerile irradiation dose on the developmental period of the F1 generation of *Grey Tea Geometridae*

[0129] Table 3: Oviposition and Fertility Parameters of F1 Generation of *Grey Tea Geometridae* after X-ray Substerile Irradiation Dose Note: Total reproductive capacity = total number of eggs laid / number of females; Daily average reproductive capacity = total number of eggs laid / number of females / interoviposition period

[0130] Table 4: Population parameters of F1 generation of *Grey Tea Geometridae* after X-ray substerile irradiation dose. Note: Net reproductive rate = Number of offspring / Total number of individuals

[0131] The results show:

[0132] Using the F1 generation produced by mating normal males with normal females as a control, we studied the F1 generation produced by mating irradiated males with normal females at doses of 160 Gy, 184.65 Gy, and 200 Gy.

[0133] (1) As shown in Table 2:

[0134] The lifespan of unirradiated F1 generation adults (5.96 days) was significantly longer than that of adults irradiated with 184.65 Gy and 200 Gy (5.47 days and 4.32 days, respectively). The sex ratio of irradiated tea geometrid moth F1 generation was significantly unbalanced. The male-to-female ratio of unirradiated F1 generation was 1.02, while the ratios at irradiation doses of 160 Gy, 184.65 Gy, and 200 Gy were 1.71, 1.88, and 2.14, respectively, significantly higher than that of the unirradiated F1 generation.

[0135] (2) As shown in Table 3:

[0136] The total reproductive capacity (272.86, 14.55, and 26.86, respectively) and daily average reproductive capacity (65.86, 3.86, and 12.53, respectively) of the F1 generation irradiated with 160 Gy, 184.65 Gy, and 200 Gy were significantly lower than those of the unirradiated F1 generation, indicating a clear downward trend in the reproductive capacity of the irradiated F1 generation.

[0137] (3) As shown in Figure 4, the survival rate curve based on age characteristics indicates:

[0138] At 35 days, the survival rate of the unirradiated F1 generation of the tea geometrid moth began to decline rapidly (Figure 4(A)); the survival rate of the F1 generation irradiated with a dose of 184.65 Gy began to decline rapidly at 9 days (larval stage) and 34 days (adult stage) (Figure 4(C)); and the survival rate of the F1 generation irradiated with doses of 160 Gy and 200 Gy began to decline rapidly at 8 days (Figure 4(B) and (D)).

[0139] Age-stage fertility curves show:

[0140] The unirradiated F1 generation population of the tea geometrid moth exhibited five reproductive peaks during the oviposition period (Figure 4(A)), while the F1 generation populations irradiated with doses of 160 Gy and 184.65 Gy exhibited four reproductive peaks during the oviposition period (Figure 4(B) and (C)).

[0141] The peak oviposition of the unirradiated F1 generation of the tea geometrid moth was 138 eggs at 37 days (Figure 4(A)); while the peak oviposition of the F1 generation of the tea geometrid moth irradiated with 160 Gy was 95 eggs at 36 days (Figure 4(B)), the peak oviposition of the F1 generation of the tea geometrid moth irradiated with 184.65 Gy was 6 eggs at 30 days (Figure 4(C)), and the peak oviposition of the F1 generation of the tea geometrid moth irradiated with 200 Gy was 11 eggs at 37 days (Figure 4(D)). The initial oviposition date of the unirradiated F1 generation population was 27 days, and the oviposition period was 16 days (Figure 4(A)); while the initial oviposition date of the F1 generation of the tea geometrid moth irradiated with a dose of 160 Gy was 29 days, and the oviposition period was 12 days (Figure 4(B)), the initial oviposition date of the F1 generation of the tea geometrid moth irradiated with a dose of 184.65 Gy was 29 days, and the oviposition period was 14 days (Figure 4(C)), and the initial oviposition date of the F1 generation of the tea geometrid moth irradiated with a dose of 200 Gy was 35 days, and the oviposition period was 6 days (Figure 4(D)).

[0142] As shown in Table 4:

[0143] The intrinsic growth rate (0.12, 0.03, and 0.02), cyclic growth rate (1.13, 1.03, and 1.01), and net reproductive rate (76.4, 2.47, and 1.88, respectively) of the irradiated F1 generation of *Tea geometrid moth* were all significantly lower than those of the irradiated F1 generation. There was no significant difference in generation cycle between the unirradiated F1 generation and the F1 generation irradiated with doses of 160 Gy and 184.65 Gy.

[0144] The results show that irradiation has a significant impact on the F1 generation of the male parent of the tea geometrid moth. The sub-sterile irradiation dose of 160 Gy and 184.65 Gy provides a guarantee for the ecological safety of field release.

[0145] The optimal dose needs to balance the relationship between the physiological indicators and sterility rate of adult tea geometrid moths after irradiation. Based on the above research results, we comprehensively analyzed the malformation rate and corrected mortality rate of irradiated tea geometrid moth pupae, as well as the emergence time, adult size, mating, sterility rate and offspring population of adults after emergence, and found that the optimal irradiation dose is 184.65 Gy.

[0146] According to the fitting calculation, the fitting equation for the sterility rate of female tea geometrid moths is y=1.012 / [1+e^(2.310-0.033x)]. It can be seen from the fitting equation that under the optimal irradiation dose of 184.65Gy, the sterility rate of female tea geometrid moths is greater than 99%, which can achieve the effect of complete sterility. This result also provides theoretical support for the subsequent simultaneous release of male and female geometrid moths in the field.

[0147] Example 3: Gray Tea Looper 60 Screening of Co-γ ray substerile irradiation dose

[0148] To determine whether X-rays are related to 60 Co-γ rays have the same sterilization effect, and we performed the procedure as described in Example 1. 60 Screening of Co-γ ray substerile irradiation dose.

[0149] The specific steps are as follows:

[0150] 1. Irradiation experiment

[0151] (1) Raising pupae:

[0152] Feed the larvae of the gray tea geometrid moth with fresh tea branches. Select 5-day-old larvae and place them in a rearing box. Record the pupation date after the larvae pupate.

[0153] (2) Irradiation:

[0154] On the second day after the larvae pupate, the sex of the pupae is distinguished under a stereomicroscope. Female and male pupae of the gray tea geometrid moth with a pupal stage of 5-6 days (late pupal stage) are selected and subjected to irradiation treatment respectively.

[0155] Nine treatment groups (40, 80, 120, 160, 200, 240, 280, 320, 360 and 400 Gy) were designed for male and female pupae according to the irradiation dose, with 25 pupae in each group and two groups (male and female) in each treatment;

[0156] The irradiation procedure was as follows: the pupae were placed in a self-sealing bag for irradiation treatment, with a wad of moist cotton placed inside to maintain humidity; the pupae in the control group were not irradiated, and the irradiation dose rate was 1.0–1.2 Gy / min.

[0157] (3) Feathering:

[0158] The irradiated pupae were placed in 1000mL packaging boxes containing moist fine sand and their eclosion was observed.

[0159] 2. Experimental Results

[0160] The deformity and corrected mortality rates, as well as the sterility rate, of pupae at each dose gradient were calculated. A logistic regression model was used to fit the sterility rate of *Tea geometrid moth* after irradiation, and a sterility rate fitting equation was obtained. Then, based on the sterility rate fitting equation, the sub-sterile irradiation dose of *Tea geometrid moth* was determined, and sterile adults of *Tea geometrid moth* were obtained using the sub-sterile irradiation dose. The results are shown in Figures 5, 6, and 7, and Table 5.

[0161] Table 5: By different 60 Summary of a two-way ANOVA of the effects of Co-γ ray irradiation dose and sex on the physiological parameters of the tea geometrid moth after irradiation

[0162] The results show:

[0163] (1) Feathering deformity rate: 60 When the Co-γ irradiation dose reaches 360 Gy and 400 Gy, the distortion rate increases significantly, while the irradiation dose of 40-320 Gy has no effect on the distortion rate (A in Figure 5).

[0164] Corrected mortality rate: Irradiation treatment with 360 Gy and 400 Gy of 60Co-γ rays had a significant effect on the corrected mortality rate of the pupa of the tea geometrid moth (B in Figure 5).

[0165] (2) 60 Co-γ ray irradiation significantly affected the reproductive parameters and adult lifespan of the brown geometrid moth of different sexes.

[0166] exist 60 Following Co-γ irradiation, the emergence time of the pupa of the gray tea geometrid moth was affected by different irradiation doses (F... 10,418 =38.17, P<0.0001, Table 5, Figure 6 (A), different genders (F) 1,418 =371.1, P<0.0001, Table 5, Figure 6 (A), interaction between irradiation dose and sex (F 10,418 = 5.060, P < 0.0001, significant effect of A) in Table 5 and Figure 6. 60 Under Co-γ ray irradiation, the lifespan of adult tea geometrid moths is affected by the irradiation dose (F). 10,418=5.167, P<0.0001, Table 5, Figure 6 (B), sex (F) 1,418 =19.39, P<0.0001, Table 5, Figure 6 (B) shows a significant effect, but 60 The interaction between Co-γ ray irradiation dose and sex had no significant effect on the lifespan of adult *Tea geometrid moth*. Regarding the mating rate of irradiated adult *Tea geometrid moth*, only... 60 The radiation dose of Co-γ rays (F 10,10 =11.20, P=0.0004, Table 5, Figure 6 (C) had a significant effect. Irradiation dose (F 10,418 =28.49, P<0.0001, Table 5, Figure 6 (D), sex (F) 1,418 =5.287, P=0.0220, Table 5, Figure 6 (D) and the interaction between irradiation dose and sex (F) 10,418 =2.009, P=0.0311, Table 5, Figure 6, D) for the recipient 60 Co-γ ray irradiation significantly affects the egg production of adult females of the tea geometrid moth.

[0167] (3) 60 The radiation dose of Co-γ rays (F 10,418 =749.1, P<0.0001) and the sex of the gray tea geometrid moth (F 1,418 =915.4, P<0.0001) both had a significant impact on the sterility rate of the irradiated tea geometrid moth.

[0168] when 60 After Co-γ ray irradiation doses reached 360 Gy, the sterility rate of female *Tea geometrid moth* reached 100%. Pairing irradiated adult *Tea geometrid moth* with unirradiated adult *Tea geometrid moth* of the opposite sex, a logistic regression model could be used to fit the relationship between irradiation dose and sterility rate well (R0). 2 >0.95).

[0169] When normal female adult tea geometrid moths are paired with irradiated male adult tea geometrid moths, the fitted curve equation for the sterility rate is y = 1.01 / [1 + e^(3.156 - 0.014x)]. Based on the calculation method of the inflection point of the logistic regression model curve, theoretically, with increasing irradiation dose, the male pupa... 60 The starting point of the rapid increase in sterility rate after Co-γ irradiation was 113.09 Gy, and the ending point was 328.31 Gy; for male pupae... 60 A Co-γ ray irradiation dose of 314.23 Gy can achieve a good sub-sterility rate (>80%).

[0170] When normal male adult tea geometrid moths are paired with irradiated female adult tea geometrid moths, the fitted curve equation for the sterility rate is y = 1.014 / [1 + e^(3.267 - 0.028x)]. Based on the calculation method of the inflection point of the logistic regression model curve, theoretically, with increasing irradiation dose, the female pupa... 60 The starting point of the rapid increase in sterility rate after Co-γ irradiation was 60.75 Gy, and the ending point was 174.28 Gy; for female pupae... 60 A Co-γ ray irradiation dose of 164.95 Gy can achieve a good sub-sterile rate (>80%); female pupae 60 A Co-γ ray irradiation dose of 251.32 Gy can achieve a good sterility rate (>99%).

[0171] Based on the above results, it can be concluded that X-rays and 60 Co-γ rays have the same sterilization effect, but 60 A Co-γ irradiation dose of 314.23 Gy is required for male *Gnaphalium affine* to achieve a good sub-sterility rate (>80%). Therefore, X-ray irradiation efficiency is significantly superior, with lower energy consumption. Furthermore, compared to cobalt sources, the accelerator in an X-ray irradiation device is controllable and source-free, offering advantages such as simple operation and no safety hazards. Based on these conclusions, we will conduct further research using X-rays as the irradiation source.

[0172] Example 4:

[0173] In the implementation of insect sterility technology, the differences in the development speed of individual insects lead to dispersed pupal formation times, making it difficult to collect pupae within the same pupal stage. This affects the efficiency of irradiation treatment and the acquisition of sterile insects. Furthermore, if adult emergence times are dispersed during field release, it is also detrimental to achieving the desired control effect. Therefore, this embodiment verifies the method of regulating insect pupal emergence time through low-temperature diapause described in this invention.

[0174] (1) Collection of pupae

[0175] The first day (D1) was recorded as the day the gray tea geometrid moth was first observed to pupate, followed by the second day (D2), the third day (D3), and the fourth day (D4). All pupae that emerged on each day were collected, and this collection continued for four days. The pupae collected on D1 and D2 were considered early-stage pupae, while those collected on D3 and D4 were considered late-stage pupae.

[0176] (2) Low temperature retardation treatment

[0177] These pupae were divided into five groups, with each group containing 50 early-stage pupae and 50 late-stage pupae.

[0178] Treatment group-1: The pupae collected on D1 and D2 were treated at 4℃ for 24h and then cultured together with the pupae collected on D3 and D4 at 25℃.

[0179] Treatment group-2: The pupae collected on D1 and D2 were treated at 4℃ for 48h and then cultured together with the pupae collected on D3 and D4 at 25℃.

[0180] Treatment group-3: The pupae collected on D1 and D2 were treated at 10℃ for 24h and then cultured together with the pupae collected on D3 and D4 at 25℃.

[0181] Treatment group-4: The pupae collected on D1 and D2 were treated at 10℃ for 48h and then cultured together with the pupae collected on D3 and D4 at 25℃.

[0182] Treatment group-5: The pupae collected on D1 and D2 were cultured together with the pupae collected on D3 and D4 at 25°C.

[0183] (3) Restore culture

[0184] After the low-temperature diapause treatment, the early-stage pupae treated with low temperature and the late-stage pupae that were not treated were transferred together to an artificial climate chamber for recovery culture. The recovery culture conditions were set as follows: temperature 25℃, relative humidity 60-70%, and photoperiod 16L:8D.

[0185] (4) Irradiation treatment

[0186] The above groups were subjected to X-ray irradiation (184.65 Gy) according to the optimal irradiation dose in Example 1;

[0187] The irradiation procedure was as follows: the pupae were placed in a self-sealing bag for irradiation treatment, with a wad of moist cotton placed inside to maintain humidity; the irradiation dose rate was 1.0–1.2 Gy / min.

[0188] Record the emergence rate of sterile pupae in each group;

[0189] 2. Experimental Results

[0190] The results are shown in Table 13.

[0191] Table 13 Effects of different low-temperature diapause conditions on the emergence time distribution of pupae of the tea geometrid moth.

[0192] As shown in Table 13, the pupae of the treatment group -5, which did not undergo low-temperature diapause treatment, had a longer eclosion process due to different pupation times, and the eclosion time was relatively dispersed, with the entire eclosion process taking 4 days to complete.

[0193] After low-temperature diapause treatment, the development process of the early-stage pupae was effectively delayed, their emergence time was increased, and their emergence time significantly overlapped with that of the untreated late-stage pupae, thus achieving concentrated emergence.

[0194] Treatment at 10℃ for 24 hours had no significant effect on the concentration of eclosion, while treatment at 10℃ for 48 hours shortened the eclosion duration to 3 days.

[0195] When treated at 4℃ for 24 hours, the eclosion duration can also be shortened to 3 days;

[0196] When treated at 4℃ for 48 hours, the eclosion process was further concentrated to be completed in 2 days, with the most significant concentration effect, and it had no significant impact on the eclosion rate of pupae.

[0197] Therefore, the 4℃, 48h treatment shows a better balance between synchronicity of emergence and survival safety, and is suitable for collecting more pupae with relatively concentrated emergence times.

[0198] Example 5: A method for precisely controlling the emergence time of insect pupae after irradiation

[0199] In the implementation of insect sterility technology, the pupal stage cannot be concentrated due to individual insect growth differences, making the collection of pupae at the same stage difficult and severely affecting irradiation efficiency and the acquisition of sterile insects. Furthermore, releasing sterile insects in the wild requires relatively concentrated emergence to achieve better control effects. To improve the operability of insect sterility technology, this invention proposes a method for precisely controlling the emergence time of insect pupae.

[0200] The specific steps are as follows:

[0201] 1. Irradiation experiment

[0202] (1) Select 3-day-old pupae of the gray tea geometrid moth with good development (normal appearance and no damage) and group them into 4 groups of 20 each. Three groups were placed in a 4℃ refrigerator for low temperature treatment for 24h, 48h and 72h respectively. The other group was not treated and was placed at room temperature (25℃) as the control group. Each treatment was repeated 3 times.

[0203] (2) The above groups after low-temperature treatment were irradiated with X-rays at the optimal irradiation dose (184.65 Gy) according to Example 1;

[0204] The irradiation procedure was as follows: the pupae were placed in a self-sealing bag for irradiation treatment, with a wad of moist cotton placed inside to maintain humidity; the pupae in the control group were not irradiated, and the irradiation dose rate was 1.0–1.2 Gy / min.

[0205] Record the emergence rate of sterile pupae in each group;

[0206] 2. The experimental results are shown in Table 6:

[0207] Table 6: Effects of low-temperature storage on pupal eclosion rate after irradiation

[0208] The results show:

[0209] Table 6 shows that there was no significant difference in the emergence rate between the control group and the group treated at 4℃ for 24 h and 48 h.

[0210] The average eclosion rate was 76.67±1.67 under treatment at 4℃ for 24 hours and 71.67±1.67 under treatment at 48 hours; while the average eclosion rate of the control group was 81.67±4.41; there was no significant difference among the three. However, the average eclosion rate was 60.00±2.89 under treatment at 4℃ for 72 hours, which was significantly different from the average eclosion rate of the control group.

[0211] Treatment at 4℃ for 48 hours had no significant effect on the emergence rate of pupae compared to the control group. Therefore, treatment at 4℃ for 48 hours was chosen as the condition for cold storage diapause, in order to facilitate the collection of more pupae and regulate the emergence time of pupae.

[0212] Example 6: A method for field release of sterile insects

[0213] To enable sterile insects to effectively suppress field pest populations, this invention proposes a method for releasing sterile insects into the field.

[0214] The specific steps are as follows:

[0215] 1. Irradiation experiment

[0216] (1) Raising pupae:

[0217] Feed the larvae of the gray tea geometrid moth with fresh tea branches. Select 5-day-old larvae and place them in a rearing box. Record the pupation date after the larvae pupate.

[0218] (2) Irradiation:

[0219] Pupae of the tea geometrid moth were collected, and low-temperature treatment was applied to pupae at different stages to regulate the emergence period. Pupae in the late pupal stage were selected, placed in a 4°C refrigerator for 48 hours, and then irradiated with an irradiation dose of 184.65 Gy. Approximately 300 pupae were treated each time.

[0220] The irradiation procedure was as follows: the pupae were placed in a self-sealing bag for irradiation treatment, with a wad of moist cotton placed inside to maintain humidity; the irradiation dose rate was 1.0–1.2 Gy / min.

[0221] 2. Field experiments

[0222] (1) Selection of tea gardens

[0223] The irradiated pupae of the tea geometrid moth obtained in step 1 were placed in tea gardens affected by the tea geometrid moth (named treatment group-1, i.e., RT-1). Another adjacent tea garden with similar damage was selected as the control group (CK group). The control group did not release sterile tea geometrid moths. The area of ​​both tea gardens was 25m*25m.

[0224] The insect population was investigated twice a week in two tea gardens using the quadrat method and the basin tapping method. Ten quadrats were set up in both the control and treatment groups, each quadrat consisting of a 1×3 meter tea row, and were numbered. The insect population counts in RT-1 and the control (CK group) are shown in Table 7. The investigation was conducted on September 5.

[0225] Table 7: Survey of insect population before release

[0226] (2) The pupae of the gray tea geometrid moth released radiation into the tea garden

[0227] The irradiated tea geometrid moths obtained in step 1 were released into the RT-1 tea garden. The first release was on September 8, and the second release was on September 22. The control group (CK group) was not exposed to tea geometrid moths.

[0228] The areas outside the tea garden are designated as numbers 1-5, and the areas inside are designated as numbers 6-10. The insect population was investigated using the basin sampling method at different times. Three samples were taken from each sample plot, and the number of insects in each sample plot was recorded.

[0229] 3. The experimental results are shown in Table 8 and Figure 8.

[0230] Table 8: Survey of insect population size after initial release

[0231] Note: ① = September 18; ② = September 22; ③ = September 25; ④ = September 28; ⑤ = October 2; ⑥ = October 6; ⑦ = October 9; ⑧ = October 13; ⑨ = October 16; ⑩ October 20; October 23; CK = control group; RT-1 = simultaneous release of sterile males and females.

[0232] (1) As shown in Table 7, there was no statistically significant difference in the number of insects between the control group (CK) and the treatment group (RT-1) (P = 0.4445, P > 0.05). The average number of insects in the control group was 26.40 ± 1.38, and that in the treatment group was 24.90 ± 1.35. The number of insects in the two groups was close, which indicates that the damage to the gray tea geometrid moth in the two tea gardens was similar to the actual number of insects, which meets the experimental requirements.

[0233] (2) As shown in Table 8, the initial survey results on September 18 after the release of the sterile tea geometrid moth showed a significant difference in insect population between the control group (CK) and the treatment group (RT-1) (P = 0.0089). During the period from September 22 to October 9, the survey results showed that the difference in insect population between the two groups was extremely significant (P < 0.0001), further confirming that the treatment measures had a significant inhibitory effect on the insect population.

[0234] (3) Due to the different timing of tea garden cleanup, the survey results on October 13 showed that the insect population in the treatment group (RT-1) was significantly higher than that in the control group (CK). After the tea garden was completely cleaned up, the surveys on October 13 and thereafter showed that the insect population in the control group (CK) was still significantly higher than that in the treatment group (RT-1) (P = 0.0218), indicating that agricultural activities had little impact on the experimental results. By October 20, the annual life cycle of the tea geometrid moth was nearing its end, and the insect populations in the two groups tended to be equal.

[0235] (4) As shown in Figure 8, at the beginning of the survey (September 18), the insect populations of the two groups were similar, with slight differences. Over time, the insect populations of both the control group (CK) and the treatment group (RT-1) increased, but the insect population of the treatment group (RT-1) remained consistently lower than that of the control group. The difference in insect populations between the two groups was extremely significant, indicating that the release of sterile insects had a significant inhibitory effect on insect populations.

[0236] Example 7: Effects of different release methods on the population suppression of the tea geometrid moth in the field

[0237] In the past, researchers only released irradiated male pupae into the wild to suppress pest populations. Based on the above research results, this invention proposes a field release method that releases both irradiated female and male pupae together. To determine the suppression effect of the two release methods on the field population of the tea geometrid moth, we compared the suppression effects of the two release methods. The specific implementation method described below is the same as in Example 5, except that after pupation, only the irradiated male pupae are released into the wild.

[0238] The specific steps are as follows:

[0239] 1. Irradiation experiment

[0240] (1) Raising pupae:

[0241] Feed the larvae of the gray tea geometrid moth with fresh tea branches. Select 5-day-old larvae and place them in a rearing box. Record the pupation date after the larvae pupate.

[0242] (2) Irradiation:

[0243] Male pupae of the tea geometrid moth were collected, and low-temperature treatment was applied to pupae at different stages to regulate the emergence period. Male pupae of the tea geometrid moth in the late pupal stage were selected, placed in a refrigerator at 4°C for 48 hours, and then irradiated with an irradiation dose of 184.65 Gy. Approximately 200 male pupae of the tea geometrid moth were treated each time.

[0244] The irradiation procedure was as follows: male pupae were placed in a self-sealing bag for irradiation treatment, with a wad of moist cotton placed inside to maintain humidity; the irradiation dose rate was 1.0–1.2 Gy / min, and the irradiation time was about 3 hours.

[0245] 2. Field experiments

[0246] (1) Selection of tea gardens

[0247] The irradiated male pupae of the tea geometrid moth obtained in step 1 were placed in tea gardens affected by the tea geometrid moth (named treatment group-2, RT-2), while treatment group-1 (RT-1) was used as a control. The area of ​​both tea gardens was 25m*25m.

[0248] The insect population was investigated twice a week in two tea gardens using the quadrat method and the basin tapping method. Ten quadrats were set up in both the control and treatment groups. Each quadrat was a 1×3 meter tea row and was numbered. The insect population counts in RT-1 and RT-2 are shown in Table 9. The investigation was conducted on September 5.

[0249] Table 9: Number of insects before release

[0250] (2) The pupae of the gray tea geometrid moth released radiation into the tea garden

[0251] The irradiated gray tea geometrid moths obtained in step 1 were released into the RT-2 tea garden. The first release was on September 8, and the second release was on September 22.

[0252] Following the method of Example 5, the irradiated gray tea geometrid moth obtained in step 1 of Example 5 was released into the RT-1 tea garden. The first release was on September 8, and the second release was on September 22.

[0253] The areas outside the tea garden are designated as numbers 1-5, and the areas inside are designated as numbers 6-10. The insect population survey was conducted using the basin sampling method, with 3 basins sampled for each sample plot. The number of insects in each sample plot was recorded at different times.

[0254] 3. The experimental results are shown in Table 10 and Figure 9.

[0255] Table 10: Number of insects after release

[0256] Note: ① = September 18; ② = September 22; ③ = September 25; ④ = September 28; ⑤ = October 2; ⑥ = October 6; ⑦ = October 9; ⑧ = October 13; ⑨ = October 16; ⑩ October 20; October 23; RT-1 = Sterile males and females released simultaneously; RT-2 = Sterile males released alone.

[0257] (1) As shown in Table 9, there was no statistically significant difference in the number of insects between treatment group-1 (RT-1) and treatment group-2 (RT-2) (P = 0.4495, P > 0.05). The average number of insects in treatment group-1 (RT-1) was 24.90 ± 1.35, and the average number of insects in treatment group-2 (RT-2) was 24.90 ± 1.35. The number of insects in the two groups was close, which indicates that the damage to the gray tea geometrid moth in the two tea gardens was similar to the actual number of insects, which meets the experimental requirements.

[0258] (2) As shown in Table 10 and Figure 9, there were slight fluctuations in the insect population of treatment group-1 (RT-1) and treatment group-2 (RT-2), but statistical analysis showed no statistically significant difference between the two groups. This indicates that the effect of releasing both male and female sterile tea geometrid moths simultaneously is basically the same as the effect of releasing only male sterile tea geometrid moths. Therefore, combining the results of these two treatment groups, it can be concluded that the control effect of releasing both male and female sterile moths simultaneously is the same as the effect of releasing male sterile tea geometrid moths alone in controlling tea geometrid moths.

[0259] Example 8: A method for assessing the quality of damaged fresh leaves

[0260] To evaluate the production and economic benefits of this method, this invention proposes a method for assessing the quality of damaged fresh leaves.

[0261] The specific steps are as follows:

[0262] (1) In the tea gardens of the control group (CK) and treatment group (RT-1, RT-2) in Examples 5 and 6, the quadrat method was used to randomly select ten quadrats, each with an area of ​​0.5*0.5 meters;

[0263] (2) Collect whole fresh leaves that have not been eaten by inchworms and can be put into actual production on October 20, and collect all fresh leaves above the upper part of the sample plot on October 23.

[0264] (3) After collection, weigh and record the data in a timely manner, and use one-way ANOVA to analyze the damage of fresh leaves in the treatment group and the control group;

[0265] (4) Experimental results are shown in Tables 11 and 12.

[0266] Table 11: Quality Assessment of Damaged Fresh Leaves

[0267] Table 12: Total weight of fresh leaves

[0268] The results show:

[0269] (1) As shown in Table 11, the average fresh leaf weight of the control group (CK) was 0.058±0.005 kg, which was significantly lower than that of the treatment groups (RT-1 and RT-2), indicating that the fresh leaf weight was low without treatment. The average fresh leaf weight of treatment group-1 (RT-1) was **0.245±0.006 kg; the average fresh leaf weight of treatment group-2 (RT-2) was 0.252±0.006 kg, which was significantly higher than that of CK, indicating that the treatment had a significant promoting effect on the fresh leaf weight. The fresh leaf weight of treatment group-2 (RT-2) was slightly higher than that of treatment group-1 (RT-1), but the difference was not statistically significant (the letters were the same, both were "b"), but it was still significantly higher than that of CK.

[0270] (2) As shown in Table 12, there were significant differences in the fresh leaf weight between the control group (CK) and treatment groups-1 (RT-1) and-2 (RT-2). The fresh leaf weight of the control group (CK) fluctuated between 0.085 kg and 0.160 kg, while the fresh leaf weight of the treatment groups was significantly higher. The fresh leaf weight of treatment group-1 (RT-1) ranged from 0.226 kg to 0.318 kg, and the fresh leaf weight of treatment group-2 (RT-2) ranged from 0.235 kg to 0.320 kg. Through one-way ANOVA, the fresh leaf weight of treatment groups-1 (RT-1) and treatment group-2 (RT-2) was significantly higher than that of the control group (CK).

[0271] Therefore, it can be concluded that the quality of fresh leaves in the treatment group was significantly higher than that in the control group, and this difference was statistically significant. This indicates that releasing sterile insects had a positive impact on increasing fresh leaf yield and economic benefits.

[0272] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A method for controlling the tea geometrid moth without the need for pupation, characterized in that, The method includes the following steps: (1) Raise the larvae of the gray tea geometrid moth to obtain 5-7 day old gray tea geometrid moth pupae. Place the 5-7 day old gray tea geometrid moth pupae in a refrigerator at 4-8℃ for 24-72 hours. (2) The pupae of the tea geometrid moth were irradiated at an irradiation dose of 160–200 Gy to obtain irradiated pupae; (3) The irradiated pupae obtained in step (2) are released directly into their natural population without the need for pupalization, and compete with wild male or female insects to mate with wild female or male insects, so that the wild female insects do not lay eggs or the eggs they lay do not hatch or reduce the number of eggs laid.

2. The method according to claim 1, characterized in that, Five- to seven-day-old pupae of the gray tea geometrid moth were placed in a 4°C refrigerator for 48 hours for low-temperature treatment before being subjected to irradiation.

3. The method according to claim 1 or 2, characterized in that, The irradiation procedure is as follows: the pupae are placed in a self-sealing bag for irradiation treatment, with a wad of moist cotton placed inside to maintain humidity; the irradiation dose rate is 1.0 to 1.2 Gy / min.

4. The method according to claim 3, characterized in that, In step (1), the method for obtaining the gray tea geometrid moth pupa is as follows: 1) Collect wild larvae of the tea geometrid moth, feed them with fresh tea branches, and raise them indoors to increase the population. 2) Collect egg masses. After the eggs hatch, transfer the newly hatched larvae of the tea geometrid moth in batches to a culture tray containing fresh tea branches. Culture them for 14 to 20 days under the conditions of an ambient temperature of 24±2℃, a humidity of 60 to 70%, and a photoperiod of 16L:8D until the larvae reach maturity. 3) Mark the pupation date after the mature larvae pupate, and collect 5-7 day old gray tea geometrid moth pupae.

5. The method according to claim 1, characterized in that, The irradiated pupae were released into their natural population 3 to 5 times.

6. The method according to claim 1, characterized in that, The irradiation dose was 184.65 Gy.

7. A method for irradiating and releasing the tea geometrid moth in a field, characterized in that, The method includes the following steps: (1) The gray tea geometrid moth was raised indoors using artificial feed breeding method to obtain gray tea geometrid moth pupae aged 5 to 7 days. The 5 to 7 day old gray tea geometrid moth pupae were placed in a refrigerator at 4 to 8℃ for low temperature treatment for 24 to 72 hours. (2) The pupae of the tea geometrid moth were irradiated at an irradiation dose of 160–200 Gy to obtain irradiated pupae; (3) Release the irradiated pupae in the field.

8. The method according to claim 7, characterized in that, Five- to seven-day-old gray tea geometrid caterpillar pupae were placed in a 4°C refrigerator for low-temperature treatment for 48 hours, followed by irradiation treatment.

9. The method according to claim 7, wherein the irradiation procedure is as follows: the pupa is placed in a self-sealing bag for irradiation treatment, and a wad of moist cotton is placed inside to maintain humidity; the irradiation dose rate is 1.0 to 1.2 Gy / min.

10. The method according to claim 7, characterized in that, In step (1), the method for obtaining the gray tea geometrid moth pupa is as follows: 1) Collect wild larvae of the tea geometrid moth, feed them with fresh tea branches, and raise them indoors to increase the population. 2) Collect egg masses. After the eggs hatch, transfer the newly hatched larvae of the tea geometrid moth in batches to a culture tray containing fresh tea branches. Culture them for 14 to 20 days under the conditions of an ambient temperature of 24±2℃, a humidity of 60 to 70%, and a photoperiod of 16L:8D until the larvae reach the mature stage. 3) Mark the pupation date after the mature larvae pupate, and collect 5-7 day old gray tea geometrid moth pupae.

11. The method according to claim 7, characterized in that, The irradiated pupae are released in the middle of the field, and the release is repeated 3 to 5 times.

12. The method according to claim 7, characterized in that, The irradiation dose was 184.65 Gy.

13. The method according to claim 7, characterized in that, The quadrat method was used to investigate the insect population in the field. Ten quadrats were set up in both the control and treatment groups. Each quadrat was a 1×3 meter tea row and was numbered. The quadrat method and the pot tapping method were used to investigate the insect population. Three pots were tapped in each quadrat.

14. The method according to claim 7, characterized in that, In the experimental fields of the control group and the treatment group, a five-point sampling method was used to select five quadrats, each with an area of ​​0.5 × 0.5 meters, and all fresh leaves from the middle and above of the tea trees in the quadrats were collected.

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