Nanometer poison bait for bactrocera dorsalis as well as preparation method and application of nanometer poison bait

Abstract

The invention belongs to the technical field of bactrocera dorsalis poison baits, and particularly relates to a bactrocera dorsalis nano poison bait and a preparation method and application thereof. The bactrocera dorsalis nano poison bait is prepared from the following raw materials in parts by weight: 1-10 parts of a microbial agent, 0.1-5 parts of a botanical agent and 0.1-5 parts of a nano carrier, the botanical medicament comprises any one of a pyrethrin medicament, an azadirachtin medicament, a rotenone medicament, a limonene medicament, a matrine medicament, a veratrine medicament and a celastrus angulatus medicament; the active probiotics in the microbial agent comprise bacillus. The bactrocera dorsalis nano poison bait provided by the invention can significantly increase the death rate of bactrocera dorsalis, contributes to a new scheme for prevention and control of bactrocera dorsalis, and contributes to green prevention and control of bactrocera dorsalis.

Description

Technical Field

[0001] The invention belongs to the technical field of citrus fruit fly poison baits, and particularly relates to a citrus fruit fly nano poison bait and a preparation method and application thereof. Background Art

[0002] The fruit fly is an important invasive pest of the family Tephritidae in the order Diptera. Bactrocera dorsalis Hendel. Its high reproductive capacity and wide host range can harm nearly 250 varieties of fruits and vegetables, including pomegranates, mangoes, citrus fruits, peaches, and pears, with multiple generations occurring annually. The fruit fly primarily infects fruit through female adults laying eggs and larvae feeding inside the fruit, causing fruit drop and rot, seriously threatening fruit yield and quality. Therefore, effective control of female fruit fly infestations is crucial for prevention and control.

[0003] Current control methods primarily include physical and chemical control. Physical control primarily involves hanging attractants in fields to trap male B. dorsalis, thereby reducing mating rates and ultimately controlling population size. Chemical control primarily involves spraying pesticides such as emamectin, deltamethrin, and trichlorfon in fields to eliminate the B. dorsalis.

[0004] However, the fruit fly has wings and can fly, so chemical pesticides sprayed in the field often fail to reach the insects and cannot effectively eliminate the females, resulting in poor control effectiveness. Furthermore, most commercially available attractants can only attract some males, and the remaining males can still reproduce by mating with females multiple times, thus failing to solve the problem of female pests, resulting in unsatisfactory practical application results. In short, current control methods for the fruit fly are all ineffective, making female fruit fly difficult to control. Therefore, there is an urgent need to develop a new strategy for controlling the fruit fly. Summary of the Invention

[0005] To address the problem that existing control methods for the citrus fruit fly are ineffective, making female citrus fruit fly difficult to control, the present invention uses female citrus fruit flies as research subjects. A microbial agent that can be used to attract female insects and a botanical pesticide, celangulin, that can be used for pest control were screened. These agents were loaded onto a nanocarrier, HLDP, to create a citrus fruit fly nanobait. The invention also provides a citrus fruit fly nanobait, its preparation method, and its use. To achieve the above objectives, the present invention employs the following technical solutions.

[0006] The first object of the present invention is to provide a nano-poison bait for the citrus fruit fly, which is made of the following raw materials in parts by weight: 1 to 10 parts of microbial agent, 0.1 to 5 parts of plant-derived agent and 0.1 to 5 parts of nano-carrier.

[0007] The plant-derived agent includes any one of pyrethrin, azadirachtin, rotenone, limonene, matrine, veratridine and celangulin.

[0008] The active probiotics in the microbial agent include Bacillus, and the concentration of the active probiotics in the microbial agent is 93%.

[0009] The nano-poison bait for citrus fruit flies of the present invention is made of a microbial agent, a botanical agent, and a nano-carrier. The microbial agent has a strong luring effect on citrus fruit flies, especially female insects. The selected botanical agent has the best poisoning effect on citrus fruit flies under the medium of the microbial agent. The nano-carrier can self-assemble with the botanical agent through hydrogen bonds and van der Waals forces to form a nano-complex, thereby enhancing the toxicity of the botanical agent to citrus fruit flies and further reducing the dosage of the agent. The prepared nano-poison bait can greatly enhance the trapping effect on citrus fruit flies, especially female insects, through the "trapping and killing" strategy, avoiding the situation where pesticide spraying cannot reach adult insects and the low trapping rate of female insects. It can solve the problem that the current control measures for citrus fruit flies in the prior art have poor control effects, resulting in the difficulty in controlling female citrus fruit flies.

[0010] Preferably, the preparation is made from the following raw materials in parts by weight: 2 to 8 parts of microbial agents, 0.5 to 3 parts of plant-derived agents, and 0.5 to 3 parts of nanocarriers.

[0011] Preferably, the preparation is made from the following raw materials in parts by weight: 3 to 6 parts of microbial agents, 0.5 to 1.5 parts of plant-derived agents, and 0.5 to 1.5 parts of nanocarriers.

[0012] Preferably, the nanocarrier is nanocarrier HLDP.

[0013] The structural formula of the nanocarrier HLDP is as follows: .

[0014] The second object of the present invention is to provide a method for preparing a nano bait for the citrus fruit fly, comprising the following steps: The microbial agent, the plant-derived agent and the nanocarrier are uniformly mixed according to the formula amount, and then incubated at room temperature for 5 minutes to 30 minutes to obtain the citrus fruit fly nano bait.

[0015] The third object of the present invention is to provide an application of a nano bait for the fruit fly in the prevention and control of the fruit fly, wherein the nano bait for the fruit fly is prepared into a nano bait solution for use.

[0016] Preferably, the mass volume concentration of the nano poison bait solution is 0.8% to 55%.

[0017] Preferably, the nano poison bait solution is applied to the surface of the prevention and control park or the isolation zone.

[0018] Preferably, the nano poison bait solution is used by spraying.

[0019] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention provides a nano-bait for the fruit fly. The nano-bait is made of a microbial agent, a botanical agent, and a nanocarrier. The botanical agent includes any one of pyrethrin, azadirachtin, rotenone, limonene, matrine, veratrine, and celangulin. The active probiotic in the microbial agent includes Bacillus. The microbial agent has a strong attractant effect on the fruit fly, especially females. The selected botanical agent has an optimal toxic effect on the fruit fly in the presence of the microbial agent. The nanocarrier can self-assemble with the botanical agent through hydrogen bonds and van der Waals forces to form a nanocomposite, thereby enhancing the toxicity of the botanical agent to the fruit fly and further reducing the dosage of the agent. The prepared nano-poison bait uses a "trapping and killing" strategy to significantly improve the effectiveness of trapping and killing the fruit fly, especially females. This avoids the situation where pesticide spraying fails to reach adult flies and the female trapping rate is low. This addresses the problem that existing control methods for the fruit fly are ineffective, making female fruit fly difficult to control. Furthermore, the nano-poison bait for the fruit fly also reduces the environmental risks associated with the excessive use of pesticides.

[0020] 2. This study used the fruit fly (Bactrocera dorsalis) as a research target. The researchers screened a nanoparticle bait containing a food-based attractant (female insects), a microbial agent, and a plant-based pesticide (ceramic acid celangulin) used as raw materials. The bait was loaded onto a nanocarrier (HLDP) to create a nanoparticle bait. This nanoparticle bait exhibits high bioactivity, combining both trapping and killing effects, significantly increasing fruit fly mortality by 20 to 40 percentage points. Furthermore, its ingredients are environmentally friendly and safe, and its spray application is convenient, contributing to the efficient and green control of the fruit fly. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is the greenhouse screening of the food source attractant for the citrus fruit fly in the present invention; wherein, Figure 1 Figure A shows the relative attraction rate of each food source attractant to the male fruit fly; Figure 1 Figure B shows the relative attraction rate of each food source attractant to female Bactrocera dorsalis; Figure 1 Figure C shows the total relative attraction rate of each food source attractant to the citrus fruit fly.

[0022] Figure 2This is the field screening of the food source attractant for the citrus fruit fly in the present invention; wherein, Figure 2 Figure A shows the relative attraction rate of each food source attractant to the male fruit fly; Figure 2 Figure B shows the relative attraction rate of each food source attractant to female Bactrocera dorsalis; Figure 2 Figure C shows the relative total attraction rate of each food source attractant to the citrus fruit fly.

[0023] Figure 3 This is the screening of plant-derived agents under the bacterial agent medium of the present invention.

[0024] Figure 4 is the structural formula of the nanocarrier HLDP in the present invention.

[0025] Figure 5 is the structural formula of celangulin in the present invention.

[0026] Figure 6 The binding mechanism of the botanical agent celangulin and the nanocarrier HLDP in the present invention is shown; wherein, "model" refers to the independent model used in the fitting model..., "variables" refers to the binding coefficient Ka, dissociation constant Kd, enthalpy change ΔH, entropy change ΔS, and "values" refers to the corresponding values ​​of the binding coefficient Ka, dissociation constant Kd, enthalpy change ΔH, and entropy change ΔS.

[0027] Figure 7 The particle size and morphology of the botanical drug celangulin before and after combining with the nanocarrier HLDP in the present invention; wherein, Figure 7 Figure A shows the particle size of the botanical drug celangulin before and after it is combined with the nanocarrier HLDP; A1 is the particle size of the botanical drug celangulin; A2 is the particle size of the botanical drug celangulin after it is combined with the nanocarrier HLDP; Figure 7 Figure B shows the morphology of the plant-derived drug celangulin before and after combining with the nanocarrier HLDP; among them, B1 is the morphology of the plant-derived drug celangulin; B2 is the morphology after combining with the nanocarrier HLDP.

[0028] Figure 8 For the biological activity evaluation of the nano bait in the present invention; wherein, Figure 8 Figure A shows the mortality rate of nano-bait to male Bactrocera dorsalis; Figure 8 Figure B shows the mortality rate of nano bait to female Bactrocera dorsalis; Figure 8 Figure C shows the total mortality rate of nano baits to the oriental fruit fly. DETAILED DESCRIPTION

[0029] The present invention will be described in detail below with reference to the accompanying drawings and specific examples, but they should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0030] Reagents and materials The reagents and materials are as follows: The preparation method of the nanocarrier HLDP is disclosed in the Chinese patent number CN 118791684 A, "A nanomaterial for preventing and treating mango anthracnose and the preparation and application of its composite", which discloses a method for preparing a nanomaterial named HLDP. The nanocarrier HLDP in the present invention is the nanomaterial named HLDP disclosed in the above patent, and its structural formula is as follows: Figure 4 shown.

[0031] The microbial agents were purchased from Yunnan Zhuoyin Biotechnology Co., Ltd.

[0032] Red date powder was purchased from Beijing Huayan Shijia Quality Inspection Technology Co., Ltd.

[0033] Banana powder was purchased from Beijing Huayan Shijia Quality Inspection Technology Co., Ltd.

[0034] Pomegranate powder was purchased from Beijing Huayan Shijia Quality Inspection Technology Co., Ltd.

[0035] Orange powder was purchased from Beijing Huayan Shijia Quality Inspection Technology Co., Ltd.

[0036] 1.5% pyrethrin emulsion in water was purchased from Yangling Fuji Biotechnology Co., Ltd.

[0037] 0.3% azadirachtin soluble solution was purchased from Beijing Qingyuanbao Biotechnology Co., Ltd.

[0038] 5% rotenone soluble solution was purchased from Yunnan Nanbao Biotechnology Co., Ltd.

[0039] 5% limonene soluble solution was purchased from Oro Agri International Co., Ltd.

[0040] 0.3% matrine aqueous solution was purchased from Yangling Fuji Biotechnology Co., Ltd.

[0041] 0.5% Veratridine soluble solution was purchased from Yangling Fuji Biotechnology Co., Ltd.

[0042] 1% celangulin aqueous emulsion was purchased from Shandong Huimin Zhonglian Biotechnology Co., Ltd.

[0043] Example 1 A nano poison bait for the citrus fruit fly is prepared from the following raw materials in parts by weight: 5 parts of microbial agents, 0.2 parts of plant-derived agents, and 0.2 parts of nano-carrier HLDP.

[0044] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0045] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0046] Take 0.2g of the nanocarrier HLDP and add 20mL of deionized water to prepare a 10mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 5g of a microbial agent and 80mL of deionized water to the nano-celangulin preparation and continue vortex mixing to prepare a 2% mass-to-volume concentration of a nano-bacterial bait for the oriental fruit fly.

[0047] Example 2 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 1 part of a microbial agent, 0.1 part of a plant-derived agent, and 0.1 part of a nano carrier HLDP.

[0048] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0049] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0050] Take 0.1g of the nanocarrier HLDP and add 10mL of deionized water to prepare a 10mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 1g of the microbial agent and 90mL of deionized water to the nano-celangulin preparation and continue vortex mixing to prepare a 1% mass-to-volume concentration of the fruit fly nanobait.

[0051] Example 3 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 10 parts of microbial agents, 5 parts of plant-derived agents, and 5 parts of nano-carrier HLDP.

[0052] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0053] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0054] Take 5.0g of the nanocarrier HLDP and add 100mL of deionized water to prepare a 50mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio. Vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 10g of a microbial agent to the nano-celangulin preparation and continue vortex mixing to prepare a 50% mass-to-volume concentration of a nano-bacterial bait for the oriental fruit fly.

[0055] Example 4 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 5.5 parts of microbial agents, 2.4 parts of plant-derived agents, and 2.4 parts of nano-carrier HLDP.

[0056] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0057] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0058] Take 2.4g of the nanocarrier HLDP and add 100mL of deionized water to prepare a 24mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 5.5g of a microbial agent to the nano-celangulin preparation and continue vortex mixing to prepare a 24% mass-to-volume concentration of a nano-bacterial bait for the oriental fruit fly.

[0059] Example 5 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 2 parts of microbial agents, 0.5 parts of plant-derived agents, and 0.5 parts of nano-carrier HLDP.

[0060] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0061] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0062] Take 0.5g of the nanocarrier HLDP and add 100mL of deionized water to prepare a 5mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 2g of a microbial agent to the nano-celangulin preparation and continue vortex mixing to prepare a 5% concentration of a nano-bacterial bait for the oriental fruit fly.

[0063] Example 6 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 8 parts of microbial agents, 3 parts of plant-derived agents, and 3 parts of nano-carrier HLDP.

[0064] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0065] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0066] Take 3.0g of the nanocarrier HLDP and add 100mL of deionized water to prepare a 30mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 8g of a microbial agent to the nano-celangulin preparation and continue vortex mixing to prepare a 30% mass-to-volume concentration of a nano-bacterial bait for the oriental fruit fly.

[0067] Example 7 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 4 parts of microbial agents, 1.5 parts of plant-derived agents, and 1.5 parts of nano-carrier HLDP.

[0068] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0069] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0070] Take 1.5g of the nanocarrier HLDP and add 100mL of deionized water to prepare a 15mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 4g of a microbial agent to the nano-celangulin preparation and continue vortex mixing to prepare a 15% mass-to-volume concentration of a nano-bacterial bait for the oriental fruit fly.

[0071] Example 8 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 3 parts of microbial agents, 0.5 parts of plant-derived agents, and 0.5 parts of nano-carrier HLDP.

[0072] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0073] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0074] Take 0.5g of the nanocarrier HLDP and add 100mL of deionized water to prepare a 5mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 3g of a microbial agent to the nano-celangulin preparation and continue vortex mixing to prepare a 5% mass-to-volume concentration of a nano-bacterial bait for the oriental fruit fly.

[0075] Example 9 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 6 parts of microbial agents, 1.5 parts of plant-derived agents, and 1.5 parts of nano-carrier HLDP.

[0076] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0077] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0078] Take 1.5g of the nanocarrier HLDP and add 100mL of deionized water to prepare a 15mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 6g of a microbial agent to the nano-celangulin preparation and continue vortex mixing to prepare a 15% mass-to-volume concentration of a nano-bacterial bait for the oriental fruit fly.

[0079] Example 10 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 4.5 parts of microbial agents, 1 part of a plant-derived agent, and 1 part of a nano carrier HLDP.

[0080] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0081] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0082] Take 1.0g of the nanocarrier HLDP and add 100mL of deionized water to prepare a 10mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 4.5g of a microbial agent to the nano-celangulin preparation and continue vortex mixing to prepare a 10% mass-to-volume concentration of a nano-bacterial bait for the oriental fruit fly.

[0083] Example 11 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 8 parts of microbial agents, 3 parts of plant-derived agents, and 3 parts of nano-carrier HLDP.

[0084] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0085] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0086] Take 3.0g of the nanocarrier HLDP and add 100mL of deionized water to prepare a 30mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 8g of a microbial agent to the nano-celangulin preparation and continue vortex mixing to prepare a 30% mass-to-volume concentration of a nano-bacterial bait for the oriental fruit fly.

[0087] Example 12 A nano poison bait for the citrus fruit fly is composed of the following raw materials in parts by weight: 6 parts of microbial agents, 1.5 parts of plant-derived agents, and 1.5 parts of nano-carrier HLDP.

[0088] Among them, the plant-derived agent is 1% celangulin aqueous emulsion.

[0089] The above-mentioned nano-bactrocera dorsalis bait is prepared into a nano-bactrocera bait solution and then used. The specific preparation method includes the following steps:

[0090] Take 1.5g of the nanocarrier HLDP and add 100mL of deionized water to prepare a 15mg / mL HLDP aqueous solution. Add an equal amount of a 1% celangulin aqueous emulsion to the HLDP aqueous solution at a 1:1 mass ratio, vortex mix thoroughly, and incubate at room temperature for 15 minutes to obtain a nano-celangulin preparation. Next, add 6g of a microbial agent to the nano-celangulin preparation and continue vortex mixing to prepare a 15% mass-to-volume concentration of a nano-bacterial bait for the oriental fruit fly.

[0091] In order to illustrate the effect of the nano-bait for the fruit fly provided by the present invention, the following study was conducted: 4. Greenhouse screening of food source attractants for Bactrocera dorsalis In a greenhouse with a length × width × height of 16.3m × 3.25m × 7m, six food source attractants for the oriental fruit fly were preliminarily screened. The mass percentage concentrations of the six food source attractants for the oriental fruit fly when used were 5%, 15%, 15%, 15%, and 15%, respectively. Among them, the pomegranate fermentation liquid was the fermentation stock liquid.

[0092] Among them, the six food source attractants for the citrus fruit fly are: microbial agent solution, red date powder solution, banana powder solution, pomegranate powder solution, orange powder solution, and pomegranate fermentation liquid.

[0093] The mass percentage concentration of the microbial agent solution is 5%. The preparation method is to weigh 5.0 g of the microbial agent into a glass beaker filled with 100 mL of deionized water and stir with a glass rod until it is uniformly dissolved.

[0094] The mass percentage concentration of the red date powder solution is 15%. The preparation method is as follows: weigh 15.0 g of red date powder into a glass beaker filled with 100 mL of deionized water, and stir with a glass rod until uniformly dissolved.

[0095] The mass percentage concentration of the banana powder solution is 15%. The preparation method is as follows: 15.0 g of banana powder is weighed into a glass beaker filled with 100 mL of deionized water, and stirred with a glass rod until uniformly dissolved.

[0096] The mass percentage concentration of the pomegranate powder solution is 15%. The preparation method is as follows: 15.0 g of banana powder is weighed into a glass beaker filled with 100 mL of deionized water, and stirred with a glass rod until uniformly dissolved.

[0097] The orange powder solution has a mass percent concentration of 15%, and is prepared by weighing 15.0 g of orange powder into a glass beaker filled with 100 mL of deionized water, and stirring with a glass rod until the solution is uniformly dissolved.

[0098] The pomegranate fermentation liquid uses fermentation stock liquid, and the preparation method of the fermentation stock liquid is as follows: first, the pomegranate is crushed, and the pomegranate and brown sugar are placed alternately in layers in a plastic bucket with a lid, that is, a layer of pomegranate and a layer of brown sugar, wherein the mass ratio of pomegranate to brown sugar is 1:1; after the placement is completed, the bucket lid is covered, and the temperature in the bucket is increased by using the temperature of sunlight; when the temperature reaches above 40°C, water and yeast are mixed and poured into the plastic bucket, and the water surface covers the pomegranate; then the bucket is fermented in an open manner for 15 days, and the contents of the bucket are stirred every 3 days during this period, and finally the fermentation stock liquid is formed.

[0099] Among them, the yeast is Angel high-activity dry yeast, which comes from Angel Yeast Co., Ltd.

[0100] The above-mentioned microbial agent solution, red date powder solution, banana powder solution, pomegranate powder solution, orange powder solution and pomegranate fermentation liquid are collectively referred to as attractant solution.

[0101] Four 1cm diameter holes were evenly punched around the sides of 250mL transparent plastic bottles. After preparing the attractant solution, 30mL was dispensed into each bottle to form trap bottles. Each bottle was considered one treatment. Six treatments were used, with five replicates per treatment. Approximately 1,000 immature Bactrocera dorsalis flies were prepared, with a 1:1 male:female ratio. The insects were released into the greenhouse one day in advance to ensure a uniform population distribution. The trap bottles were randomly hung in the greenhouse. One day later, the number of male and female insects in each bottle was counted, and the relative attraction rate of each attractant was calculated. Based on the greenhouse test results, the top three attractants were selected for further screening in the field. Three treatments were used for field screening, with 12 replicates per treatment. Three trap bottles within each treatment were evenly distributed on the same tree, hanging at a height of approximately 1.5m. One month later, the number of male and female insects captured by each treatment was measured, and the relative attraction rate was calculated.

[0102] Among them, the sexually immature fruit fly is the fruit fly that has emerged within 7 days.

[0103] The results of greenhouse screening of food source attractants for Bactrocera dorsalis are as follows: Figure 1 shown.

[0104] Depend on Figure 1 As shown in Figure A, the relative attraction rates of the six food source attractants, bacterial agent, red date powder, pomegranate fermentation liquid, banana powder, pomegranate powder and orange powder to male citrus fruit flies are 73.03%, 10.23%, 6.51%, 5.58%, 3.72% and 0.93% respectively.

[0105] Depend on Figure 1 As shown in Figure B, the relative attraction rates of the six food source attractants, bacterial agent, red date powder, pomegranate fermentation liquid, banana powder, pomegranate powder and orange powder to female citrus fruit flies are 16.47%, 28.23%, 29.41%, 10.59%, 8.82% and 6.47% respectively.

[0106] Depend on Figure 1 As shown in Figure C, the total relative attraction rates of the six food source attractants, bacterial agent, red date powder, pomegranate fermentation liquid, banana powder, pomegranate powder and orange powder to the citrus fruit fly are 48.05%, 18.18%, 16.62%, 7.8%, 5.97% and 3.38% respectively.

[0107] According to the greenhouse screening results, the top three attractants for male, female, and total relative attraction rates of the six food-based attractants were bacterial inoculant, red date powder, and pomegranate fermentation liquid. Therefore, these three attractants were initially selected in the greenhouse screening experiment.

[0108] The results of field screening of food source attractants for Bactrocera dorsalis are as follows: Figure 2 shown.

[0109] Depend on Figure 2 As shown in Figure A, the relative attraction rates of the three food-based attractants, bacterial agent, red date powder, and pomegranate fermentation liquid to male citrus fruit flies are 81.4%, 16.28%, and 2.33%, respectively.

[0110] Depend on Figure 2 As shown in Figure B, the relative attraction rates of the three food-source attractants, bacterial agent, red date powder, and pomegranate fermentation liquid to female citrus fruit flies are 77.63%, 19.74%, and 2.63%, respectively.

[0111] Depend on Figure 2 As shown in Figure C, the total relative attraction rates of the three food-source attractants, bacterial agent, red date powder, and pomegranate fermentation liquid to the citrus fruit fly are 75%, 22.86%, and 2.14%, respectively.

[0112] Further field screening of the three attractants initially selected from the greenhouse experiments revealed that the most effective attractants, measured by male and female attraction, and overall attraction, were all bacterial agents. This resulted in the identification of a highly effective food-source attractant for female insects—a microbial agent.

[0113] 5. Research on screening of plant-derived agents under microbial agent medium To investigate which botanical agents could be combined with microbial agents to effectively trap and kill the fruit fly ( Bactrocera dorsalis), we prepared seven baits using a microbial attractant system. Seven botanical agents—1.5% pyrethrin emulsion (water), 0.3% azadirachtin soluble solution, 5% rotenone soluble solution, 5% limonene soluble solution, 0.3% matrine aqueous solution, 0.5% veratridine soluble solution, and 1% celangulin aqueous emulsion—were diluted 500-fold to create seven different baits. A stomach poisoning method was used, with eight treatments, including a single agent as a blank control. Each treatment had five replicates, with 30 fruit flies per replicate, and a 1:1 female-to-male ratio. Treatment mortality was measured every 6 hours for 48 hours.

[0114] The stomach poison method involves the following steps: Take a 0.4g cotton ball and adsorb 6mL of each bait and inoculant onto it. Place the cotton ball in the center of a 6cm petri dish. Then, place it in the center of a 30cm x 30cm x 30cm tall net cage. Place fruit flies in the cage. The flies are attracted to the bait and consume it, ultimately killing them. Each net cage is considered a replicate.

[0115] The results of screening of plant-derived agents under bacterial agent medium are as follows Figure 3 shown.

[0116] Depend on Figure 3 It can be seen that the poison baits formed by combining seven plant-derived agents, namely pyrethrin, azadirachtin, rotenone, limonene, matrine, veratridine and celangulin with bacterial agents have a mortality rate of 62%, 40%, 25%, 6%, 41%, 65% and 99% against the citrus fruit fly in the 48th hour respectively.

[0117] The bait prepared with celangulin and a microbial agent had the highest mortality rate and exhibited the highest mortality against the fruit fly at all survey time points, far exceeding the lethality of baits prepared with six other botanical pesticides and microbial agents. Thus, celangulin, a highly effective botanical insecticide in a microbial agent medium, was identified.

[0118] 3. Binding mechanism of the botanical drug celangulin and the nanocarrier HLDP Isothermal titration calorimetry was used to determine the thermodynamic parameters of the binding between the nanocarrier HLDP and celangulin to analyze the binding mechanism. The experiment was conducted using a Nano ITC instrument. 250 μL of a 0.3 mmol / L nanocarrier HLDP solution was added dropwise to 1 mL of a 0.03 mmol / L celangulin solution. The titration was performed 25 times, with 10 μL titrations each time. The reaction heat data were analyzed using NanoAnalyze software, and the Gibbs free energy change was calculated using the formula ΔG = ΔH - TΔS.

[0119] Among them, Nano ITC: TA Instruments Waters, USA.

[0120] Structural characteristics of nanocarrier HLDP: It is a linear block polymer. Its structural formula is Figure 4 shown.

[0121] Structural characteristics of celangulin: The molecular formula is C 32 H40O 14 , is a dihydroagarwood furan polyol ester compound, its structural formula is as follows Figure 5 shown.

[0122] The binding mechanism of the plant-derived drug celangulin and the nanocarrier HLDP is shown in the following results. Figure 6 shown.

[0123] Depend on Figure 6 It can be seen that the binding coefficient Ka (M -1 ) is 3.250×10 5 The dissociation constant Kd (M) is 3.077×10 -6, indicating a strong interaction between the two. The enthalpy change, ΔH, and entropy change, ΔS, of the binding reaction were -621.4 kJ / mol and -1979 J / mol·K, respectively. The calculated Gibbs free energy change, ΔG, was -31.658 kJ / mol. ΔG < 0, indicating that celangulin and the nanocarrier, HLDP, spontaneously bind through hydrogen bonds and van der Waals forces.

[0124] 4. Particle size and morphology of the botanical drug celangulin before and after combining with the nanocarrier HLDP The particle size and morphology of the botanical drug celangulin before and after binding to the nanocarrier HLDP were determined using a nanoparticle size analyzer and scanning electron microscopy. The botanical drug celangulin and the nanocarrier HLDP were vortex-mixed at a mass ratio of 1:0 and 1:1 at room temperature and incubated for 15 minutes. The final effective concentration of celangulin was 0.02 mg / mL. 3 mL of each system was placed in a quartz glass dish and placed in a nanoparticle size analyzer for particle size measurement. Simultaneously, 10 μL of each system was dripped onto a silicon wafer, dried at room temperature, and then gold-sprayed. The morphology was then observed using a scanning electron microscope.

[0125] The particle size and morphology of the plant-derived drug celangulin before and after combining with the nanocarrier HLDP are shown in the figure. Figure 7 shown.

[0126] Depend on Figure 7 It can be seen that the particle size of celangulin is 801.67 nm, and the particle size of celangulin + nanocarrier HLDP is 362.40 nm. The morphologies of the two are mainly nearly spherical under a scanning electron microscope.

[0127] The addition of nanocarrier HLDP significantly reduced the particle size of celangulin without significantly changing its morphology.

[0128] 5. Evaluation of the biological activity of nanobaits In a microbial system, commercial celangulin and the nanocarrier HLDP were added at a mass ratio of 1:0, 1:1, and 0:1, respectively, and then diluted to ensure that the celangulin was diluted to 800 times. The experiment was conducted using a stomach poisoning method with four treatments, including the microbial agent alone as a blank control. Each treatment had three replicates, with 30 fruit flies per replicate, at a 1:1 male-female ratio. Mortality was measured every 6 hours for 48 hours.

[0129] The biological activity evaluation results of nano bait are as follows Figure 8 shown.

[0130] Depend on Figure 8It can be seen that at the eight survey time points, the mortality rates of nano baits on male fruit flies were 1.33%, 5.33%, 16%, 30.67%, 54.67%, 62.67%, 76% and 89.33% respectively; the mortality rates of nano baits on female fruit flies were 2.67%, 5.33%, 17.33%, 26.67%, 48%, 65.33%, 82.66% and 89.33% respectively; the total mortality rate of nano baits on fruit flies was 2%, 5.33%, 16.67%, 28.67%, 51.33%, 64%, 79.33% and 89.33% respectively.

[0131] The nanobaits were more effective against the fruit fly than the microbial agent and the bait without the nanocarrier HLDP. After 48 hours of treatment, the nanobaits had a nearly 100% lethality against the fruit fly, demonstrating excellent biological activity.

[0132] From the above results, it can be seen that the nano-poison bait for Bactrocera dorsalis provided by the present invention has a highly effective trapping and killing effect.

[0133] The present invention screened a microbial agent that can be used to attract female citrus fruit fly and a botanical pesticide, celangulin, that can be used for prevention and control, and used a nanocarrier HLDP to load them to create a nano bait for citrus fruit fly. The bait can significantly increase the mortality rate of citrus fruit fly, contributing a new solution to fruit fly prevention and control, and facilitating green and efficient prevention and control of citrus fruit fly.

[0134] Although preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once the basic inventive concepts become known, and all such changes and modifications fall within the scope of the present invention.

Claims

1. A nano poison bait for Bactrocera dorsalis, characterized in that: The raw materials are as follows: 1 to 10 parts of microbial agent, 0.1 to 5 parts of plant-derived agent and 0.1 to 5 parts of nanocarrier. The plant-derived agent includes any one of pyrethrin, azadirachtin, rotenone, limonene, matrine, veratridine and celangulin; The active probiotics in the microbial agent include Bacillus, and the concentration of the active probiotics in the microbial agent is 93%.

2. The nano bait for Bactrocera dorsalis according to claim 1, characterized in that: The invention is prepared from the following raw materials in parts by weight: 2 to 8 parts of microbial agents, 0.5 to 3 parts of plant-derived medicines and 0.5 to 3 parts of nano-carriers.

3. The nano poison bait for Bactrocera dorsalis according to claim 2, characterized in that: The invention is prepared from the following raw materials in parts by weight: 3 to 6 parts of microbial agents, 0.5 to 1.5 parts of plant-derived agents and 0.5 to 1.5 parts of nano-carriers.

4. The nano bait for Bactrocera dorsalis according to claim 1, characterized in that: The nanocarrier is a nanocarrier HLDP; The structural formula of the nanocarrier HLDP is as follows: 。 5. The method for preparing a nano bait for Bactrocera dorsalis according to any one of claims 1 to 4, characterized in that: The steps include: The microbial agent, the plant-derived agent and the nanocarrier are uniformly mixed according to the formula amount, and then incubated at room temperature for 5 minutes to 30 minutes to obtain the citrus fruit fly nano bait.

6. Use of the Bactrocera dorsalis nano bait according to claim 1 in the prevention and control of Bactrocera dorsalis, characterized in that: The oriental fruit fly nano bait is prepared into a nano bait solution and then used.

7. The use according to claim 6, characterized in that The mass volume concentration of the nano poison bait solution is 0.8% to 55%.

8. The use according to claim 7, characterized in that The nano poison bait solution is applied to the surface of the prevention and control park or the isolation zone.

9. The use according to claim 8, characterized in that The nano poison bait solution is used by spraying.

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