Bisexual attractant of bactrocera dorsalis and preparation method and application thereof

Abstract

The application provides a bethyla huidobrensis bisexual attractant, a preparation method and application thereof. The bethyla huidobrensis bisexual attractant comprises methyl eugenol and isoamyl acetate. The attractant significantly improves the attraction effect on male and female bethyla huidobrensis, reduces the use amount of methyl eugenol, has the advantages of low cost and simple preparation, and is suitable for industrialized production of the bethyla huidobrensis killing agent.

Description

Technical Field

[0001] This invention belongs to the field of plant disease and pest control technology. Specifically, this application provides a bisexual attractant for the oriental fruit fly, its preparation method, and its application. Background Technology

[0002] The oriental fruit fly (Bactrocera dorsalis), commonly known as the yellow fruit fly or fruit maggot, belongs to the order Dipetera, family Tephritidae, subfamily Dacinae, and genus Bactrocera. It is a globally dangerous quarantine pest. Currently, this pest occurs and causes damage in Guangdong, Guangxi, Hunan, Guizhou, Fujian, Hainan, Yunnan, and Sichuan provinces of my country, and shows a trend of gradually invading and spreading to northern regions. The oriental fruit fly has a wide host range, damaging more than 250 species of fruits and vegetables from 46 families, including guava, mango, citrus, pomegranate, jujube, peach, apple, and persimmon. The female adult lays eggs inside the host fruit using her ovipositor; the larvae then feed on the eggs, causing fruit rot and drop. In severely infested areas, this insect can cause crop yield losses of over 80%, or even total crop failure, seriously affecting the quantity and quality of agricultural products in my country and causing significant losses to my country's agricultural trade.

[0003] Because the larvae bore into the fruit, their damage is concealed, making them difficult to control. The adult stage, the only time the oriental fruit fly is exposed, is the critical period for control. Adult attractants are a common method of pest control. Currently, most commercially available fruit fly attractants are factory-produced methyl eugenol (ME). However, ME attractants have some shortcomings. First, while they are effective at attracting male oriental fruit flies, they have no effect on females, and males can mate multiple times. The key to damage lies in the female's egg-laying and larval feeding. Second, not all males show a strong attraction to ME. Furthermore, ME has been found to be a toxic carcinogen and is unsuitable for long-term field use. Therefore, it is essential to enhance the attraction effect on female adults and reduce the use of ME.

[0004] Therefore, there is an urgent need to develop a novel attractant for the oriental fruit fly, enhancing its attraction to adult females while reducing the amount of metabolites (ME) used, achieving higher trapping efficiency and environmental friendliness, and thus enabling effective control of the oriental fruit fly. This patent application aims to introduce a novel attractant and its formulation, utilizing the synergistic effect of natural plant volatiles and synthetic compounds to provide excellent attraction, thereby offering a feasible solution for the control of the oriental fruit fly. Summary of the Invention

[0005] In response to the above situation, this application provides a bisexual attractant for the oriental fruit fly, which comprises methyl eugenol and isoamyl acetate.

[0006] Furthermore, the oriental fruit fly attractant contains 40-60% v / v methyl eugenol and 40-60 μL / mL isoamyl acetate.

[0007] Furthermore, the oriental fruit fly attractant is a paraffin oil solution, wherein the active ingredients are 50% v / v methyl eugenol and 50 μL / mL isoamyl acetate.

[0008] On the other hand, this application provides a method for preparing the above-mentioned oriental attractant for the oriental fruit fly. The method includes mixing 900 μL of paraffin oil and 100 μL of isoamyl acetate to obtain a 100 μL / ml isoamyl acetate solution; mixing 500 μL of the 100 μL / ml isoamyl acetate solution with 500 μL of methyl eugenol to obtain a mixed attractant containing 50 μL / ml isoamyl acetate and 50% methyl eugenol.

[0009] On the other hand, this application provides the application of the above-mentioned oriental fruit fly attractant in the preparation of pesticides or devices for trapping or killing oriental fruit flies.

[0010] Furthermore, the pesticide or device used to trap or kill oriental fruit flies is a pesticide or device used to trap or kill mated oriental fruit flies.

[0011] Furthermore, the pesticide or device used to trap or kill oriental fruit flies is a pesticide or device used to trap or kill mated female oriental fruit flies.

[0012] Furthermore, the pesticide or device used to trap or kill oriental fruit flies is a pesticide or device used to trap or kill immature female oriental fruit flies.

[0013] Furthermore, the device is a trap, a lure, or a sticky insect board.

[0014] On the other hand, this application provides a device for trapping or killing oriental fruit flies, the device comprising the above-mentioned oriental fruit fly bisexual attractant.

[0015] The attractant of the present invention has a significant attraction effect on adult oriental fruit flies, especially mated females; the components are simple and easy to obtain, the cost is low, and the preparation method is simple; it does not require changes to the original oriental fruit fly attraction device conditions, and can be used in combination with traps, lures, and sticky insect boards, making it easy to operate. Attached Figure Description

[0016] Figure 1 Comparison of selectivity of oriental fruit fly for methyleugenol and isoamyl acetate (selectivity rates are in parentheses).

[0017] Figure 2 Comparison of selectivity of oriental fruit fly to methyleugenol and mixed attractants (selectivity rate in parentheses).

[0018] Figure 3 The attraction curves of different attractants to the oriental fruit fly are shown. In the figure, part A represents immature fruit flies, part B represents sexually mature fruit flies that have not yet mated, and part C represents sexually mature fruit flies that have already mated.

[0019] Figure 4 Comparison of the total number of male and female oriental fruit flies attracted by different attractants after 3 hours. In the figure, part A represents immature fruit flies, part B represents sexually mature fruit flies that have not yet mated, and part C represents fruit flies that have already mated. Detailed Implementation

[0020] To facilitate understanding of the present invention, the technical solutions described below will be further explained in conjunction with embodiments. It should be understood that the following embodiments are provided for illustrative purposes only and are not intended to limit the scope of the invention. Those skilled in the art can make various modifications and substitutions to the present invention without departing from its spirit and intent.

[0021] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. All materials and reagents used in the following examples are commercially available.

[0022] All data in this invention were analyzed using SPSS Statistics package 20 software. Differences between different treatments were statistically analyzed using one-way ANOVA with Tukey or Games-Howell tests (significance level: P < 0.05).

[0023] The citrus fruit flies were raised in an artificially bred population at the IPM Laboratory of the Department of Entomology, College of Plant Protection, China Agricultural University. Fruit fly eggs were obtained from fresh bananas. The fresh bananas and eggs were placed together in a rearing box containing an appropriate amount of wheat bran and kept in a temperature- and humidity-controlled rearing room (temperature 27±2℃, relative humidity 60±5%, photoperiod 14L:10D). After 7-8 days, when the larvae matured and exhibited jumping behavior, moist sand was prepared for the next jumping operation. After jumping, any remaining inactive larvae were manually removed with tweezers and placed in the sand. The sand was then poured into a clean rearing box, and the box was placed in an incubator for 8-9 days to pupate. Once all larvae had pupated, the rearing box was opened and the larvae were placed in a rearing cage. Adult feed and water were prepared and placed in the cage. After emergence, adult feed and water were provided every 2 days.

[0024] The test insects were: sexually immature female adults (3-5 days old), sexually mature but unmated female adults (7-10 days old; males and females were separated and raised separately on the 3rd day after emergence until the required age for the experiment), and sexually mature and mated female adults (15-20 days old).

[0025] The reagent preparation method is as follows:

[0026] Preparation of 50 μL / ml isoamyl acetate solution: Pipette 950 μL of paraffin oil and 50 μL of isoamyl acetate into a clean centrifuge tube and mix thoroughly to obtain 50 μL / ml isoamyl acetate solution.

[0027] Preparation of 50% methyl eugenol solution: Pipette 500 μL of paraffin oil and 500 μL of methyl eugenol into a clean centrifuge tube and mix thoroughly to obtain 50% methyl eugenol solution.

[0028] Preparation of the mixed attractant: Pipette 900 μL of paraffin oil into a clean centrifuge tube, then pipette 100 μL of isoamyl acetate into it and mix thoroughly to obtain a 100 μL / ml isoamyl acetate solution; pipette 500 μL of the 100 μL / ml isoamyl acetate solution and 500 μL of methyl eugenol into a clean centrifuge tube and mix thoroughly to obtain a mixed attractant containing 50 μL / ml isoamyl acetate and 50% methyl eugenol.

[0029] Example 1: Comparison of the attraction effects of isoamyl acetate and methyl eugenol on oriental fruit flies.

[0030] Using filter paper containing 50 μL / ml isoamyl acetate solution and 50% methyl eugenol solution as odor sources, 50 male and 50 female insects at different physiological stages were used as a group for Y-type olfaction experiments.

[0031] The experiment used 5cm of the same size 2 Using filter paper as a carrier, 200 μL of the test substance solution was evenly dropped onto the filter paper. The filter paper was then placed in an odor bottle as an odor source. During the experiment, the gas flow rate was 80 mL / min. In each test, one adult insect was placed at the end of the Y-arm olfactory tube, and the behavior of the adult insect was observed after 10 minutes. The ambient temperature was 25℃, and the humidity was 60%. Each test insect was used only once.

[0032] The selection criteria are as follows: if the test insect crawls into 2 / 3 of an arm and stays there for more than 1 minute during the test, it is considered that the test insect has made a selection for the treatment substance, and the data is recorded. If the selection criteria are not met, it is recorded as no selection has been made. To eliminate any potential impact on the insects, after every 20 test insects have completed the test treatment, the Y-arm olfactory instrument is cleaned with anhydrous ethanol to remove residual odors, dried, and then the Y-arm olfactory instrument is flipped over and the test arms are exchanged to reduce errors.

[0033] The experimental results showed that, compared with 50% methyl eugenol solution, 50 μL / ml isoamyl acetate solution had a significant attraction effect on mated female adults, while its attraction effect on male adults was far less than that of methyl eugenol. The mixture had a significant attraction effect on mated female adults and also a significant attraction effect on sexually mature male adults (see...). Figure 1 ).

[0034] Example 2: Comparison of the attraction effects of a mixture of isoamyl acetate and methyl eugenol on oriental fruit flies with methyl eugenol.

[0035] The experiment used a Y-arm olfactometer to determine the selective tropism of the oriental fruit fly towards the mixed agent and methyleugenol. The specific experimental method was the same as in Example 1.

[0036] The experimental results showed that, compared with a 50% methyl eugenol solution, the mixture (i.e., an attractant containing 50 μL / ml isoamyl acetate and 50% methyl eugenol) had a significant attraction effect on mated female adults, and also had a significant attraction effect on both unmated and mated male adults (see...). Figure 2 ).

[0037] Example 3 Enclosure Experiment

[0038] The experiment compared the attraction effects of four treatments on the oriental fruit fly: a mixture of 50 μL / ml isoamyl acetate and 50% methyl eugenol, 50 μL / ml isoamyl acetate, 50% methyl eugenol, and paraffin oil. Blank lures were soaked in each of the four liquids for 2 hours. The different lures were then fixed to the center of white sticky insect boards, which were randomly and vertically suspended from the top of a 1m×1m×1m cage. The room temperature was 28℃, the relative humidity was 65±5%, and ventilation was maintained. The test insects included immature female adults, sexually mature but unmated female adults, and sexually mature and mated female adults. 100 insects were introduced each time, 50 males and 50 females. A small amount of water and feed was placed in the center of the cage floor to ensure the fruit flies' survival. The experiment was conducted daily from 12:00 to 15:00. The number of male and female insects attracted to the insect boards was recorded every 5 minutes using a camera, with three replicates.

[0039] The results showed that the mixture (i.e., attractants of 50 μL / ml isoamyl acetate and 50% methyl eugenol) had the fastest attraction rate for both male and female adults of the oriental fruit fly at all stages (see [link]). Figure 3 Within 3 hours, the total number of mated male and female insects attracted by the mixture was significantly higher than that of other treatments, and the mixture significantly improved the attraction effect on adults at all stages (see...). Figure 4 ).

[0040] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, reasonable modifications and substitutions can be made to the present invention without departing from its spirit.

Claims

1. The application of a fruit fly attractant in the preparation of pesticides or devices for trapping or killing mated female fruit flies, characterized in that, The method for preparing the oriental fruit fly attractant is as follows: 900 μL of paraffin oil and 100 μL of isoamyl acetate are mixed to obtain a 100 μL / ml isoamyl acetate solution; 500 μL of the 100 μL / ml isoamyl acetate solution is mixed with 500 μL of methyl eugenol to obtain the oriental fruit fly attractant.

2. The application according to claim 1, wherein the device is a trap, a lure, or a sticky insect board.

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