Bactrocera dorsalis dual-sex attractant as well as preparation method and application thereof

Abstract

The invention provides a bactrocera dorsalis dual-sex attractant as well as a preparation method and application thereof. The bactrocera dorsalis dual-sex attractant comprises methyleugenol and isoamyl acetate. The attractant obviously improves the attraction effect on male and female bactrocera dorsalis individuals, reduces the use amount of methyleugenol, has the advantages of low cost and simple preparation, and is suitable for industrial production of the bactrocera dorsalis attractant.

Description

Technical Field

[0001] The present invention belongs to the technical field of plant disease and insect pest control. Specifically, the present application provides a bisexual attractant for Bactrocera dorsalis and a preparation method and application thereof. Background Art

[0002] Bactrocera dorsalis, commonly known as yellow fly, fruit maggot, oriental fruit fly, belongs to Dipetera, Tephritidae, Dacinae, Bactrocera, and is a dangerous quarantine pest in the world. At present, the pest has occurred and caused damage in Guangdong, Guangxi, Hunan, Guizhou, Fujian, Hainan, Yunnan, Sichuan and other provinces and regions in my country, and is gradually invading and spreading to northern my country. Bactrocera dorsalis has a wide range of hosts and can harm more than 250 kinds of fruits and vegetables in 46 families, such as guava, mango, citrus, pomegranate, jujube, peach, apple, persimmon, etc. Its damage mode is that the female adult uses the ovipositor to insert the host fruit to lay eggs, and the larvae feed on it, causing the fruit to rot and fall off. In areas where the pest is seriously infested, it can cause more than 80% crop yield loss, or even crop failure, which seriously affects the yield and quality of my country's agricultural products and causes huge losses to my country's agricultural product trade.

[0003] Because the larvae bore into the fruit, the damage is concealed and difficult to control. As the only period when the adults are exposed to the citrus fruit fly, it becomes a critical period for prevention and control. Adult attractants are one of the commonly used methods for pest control. At present, most of the commercialized fruit fly attractant products on the market are factory-produced methyl eugenol (Methyl Eugenol, ME). However, ME attractants have some shortcomings. First, although it has a good effect on attracting male adults of the citrus fruit fly, it has no attracting effect on females, while males can mate multiple times, and females laying eggs and hatching larvae are the key to damage. Secondly, not all males have a clear tropism response to ME. In addition, ME has been found to be a toxic carcinogen and is not suitable for long-term use in the field. Therefore, it is necessary to enhance the attracting effect on female adults and reduce the use of ME.

[0004] Therefore, it is urgent to develop a new type of attractant for the orange fruit fly, enhance the attracting effect on female adults, reduce the amount of ME used, have higher trapping efficiency and environmental protection characteristics, and achieve effective control of the orange fruit fly. This patent application aims to introduce a new type of attractant and its formula, which uses the synergistic effect of natural plant volatiles and synthetic compounds to provide excellent attracting effect, thereby providing a feasible solution for the control of the orange fruit fly. Summary of the invention

[0005] In view of the above situation, on the one hand, the present application provides a bisexual attractant for the citrus fruit fly, which comprises methyl eugenol and isoamyl acetate.

[0006] Furthermore, the bisexual attractant for Bactrocera dorsalis contains 40-60% v / v methyl eugenol and 40-60 μL / mL isoamyl acetate.

[0007] Furthermore, the bisexual attractant for the oriental fruit fly 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, the present application provides a method for preparing the above-mentioned bisexual attractant for citrus fruit fly, the method comprising mixing 900 μL of paraffin oil and 100 μL of isoamyl acetate to obtain a 100 μL / ml isoamyl acetate solution; taking 500 μL of the 100 μL / ml isoamyl acetate solution and mixing it 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, the present application provides the use of the above-mentioned bisexual attractant of the citrus fruit fly in the preparation of pesticides or devices for trapping or killing the citrus fruit fly.

[0010] Furthermore, the pesticide or device for trapping or killing the fruit fly is a pesticide or device for trapping or killing mated fruit flies.

[0011] Furthermore, the pesticide or device for trapping or killing the citrus fruit fly is a pesticide or device for trapping or killing mated female citrus fruit flies.

[0012] Furthermore, the pesticide or device for trapping or killing the citrus fruit fly is a pesticide or device for trapping or killing sexually immature female citrus fruit fly.

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

[0014] On the other hand, the present application provides a device for trapping or killing the citrus fruit fly, which comprises the above-mentioned citrus fruit fly bisexual attractant.

[0015] The attractant of the invention has a significant attracting effect on adult citrus fruit flies, especially mated females; the components are simple and easy to obtain, the cost is low, and the preparation method is simple; there is no need to change the conditions of the original citrus fruit fly attracting device, and the attractant can be used in combination with a trap, an attractant core, and a sticky insect board, and the operation is simple. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Comparison of the selectivity of Bactrocera dorsalis to methyl eugenol and isoamyl acetate (the selectivity is in brackets).

[0017] Figure 2 Comparison of the selectivity of Bactrocera dorsalis to methyl eugenol and mixed attractants (the selectivity is in brackets).

[0018] Figure 3 The curve of the attractant effect of different attractants on the citrus fruit fly. Part A in the figure is immature, part B is sexually mature but not mated, and part C is mated.

[0019] Figure 4 Comparison of the total number of male and female fruit flies lured by different attractants within 3 hours. In the figure, part A is sexually immature, part B is sexually mature but not mated, and part C is mated. DETAILED DESCRIPTION

[0020] In order to make the content of the present invention easier to understand, the technical scheme of the present invention will be further described below in conjunction with embodiments. It should be understood that the following embodiments are provided only for the purpose of illustration and are not intended to limit the scope of the present invention. Those skilled in the art may make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention.

[0021] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods. The materials, reagents, etc. used in the following examples can all be obtained from commercial sources.

[0022] All statistical data of the present invention were analyzed using SPSS Statistics package 20. The differences between different treatments were analyzed using one-way analysis of variance Tukey or Games-Howell test (significance level was P<0.05).

[0023] The bred Bactrocera dorsalis was from a population artificially reared in the IPM Laboratory, Department of Entomology, College of Plant Protection, China Agricultural University. Fresh bananas were used to obtain Bactrocera dorsalis eggs, which were placed in an insect box containing an appropriate amount of wheat bran together with the fresh bananas and placed in a constant temperature and humidity insect room (temperature 27±2℃, relative humidity 60±5%, photoperiod 14L:10D) for breeding. After the mature larvae showed jumping behavior on the 7th to 8th day, moistened sand was prepared for the next jumping operation. After the jumping, the remaining inactive larvae could be manually picked up with tweezers and placed in the sand. The sand was poured into a clean insect box and placed in an incubator for 8-9 days to wait for pupation. After all pupations, the insect box was opened and placed in an insect cage. The adult feed and adult waterer were prepared and placed in the insect cage. After the emergence, the adult feed was added every 2 days and water was added for breeding.

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

[0025] The reagent preparation method is:

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

[0027] Preparation of 50% methyl eugenol solution: Use a pipette to transfer 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 mixed attractant: use a pipette to transfer 900 μL of paraffin oil into a clean centrifuge tube, then use a pipette to transfer 100 μL of isoamyl acetate into the oil, mix thoroughly to obtain a 100 μL / ml isoamyl acetate solution; take 500 μL of the 100 μL / ml isoamyl acetate solution and 500 μL of methyl eugenol into a clean centrifuge tube and mix to obtain a mixed attractant containing 50 μL / ml isoamyl acetate and 50% methyl eugenol.

[0029] Example 1 Comparison of the attractant effects of isoamyl acetate and methyl eugenol on Bactrocera dorsalis

[0030] A Y-type olfactometer experiment was conducted using filter papers containing 50 μL / ml isoamyl acetate solution and 50% methyl eugenol solution as odor sources, with 50 male and female insects of different physiological stages as groups.

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

[0032] The selection judgment conditions are as follows: if the test insect crawls into 2 / 3 of an arm and stays there for more than 1 minute during this period, it is considered that the test insect has made a choice for the treatment substance and the data is recorded. If the judgment conditions are not met, it is recorded as no choice. In order to eliminate possible influences on insects, after every 20 test insects complete the test treatment, the Y-arm olfactometer is cleaned with anhydrous ethanol to eliminate residual odor, and after drying, the Y-arm olfactometer is turned over and the test arm is exchanged to reduce errors.

[0033] The test results show that compared with 50% methyl eugenol solution, 50μL / ml isoamyl acetate solution has a significant attracting effect on mated female adults, but its attracting effect on male adults is far less than that of methyl eugenol. The mixture has a significant attracting effect on mated female adults and has a significant attracting effect on sexually mature male adults (see Figure 1 ).

[0034] Example 2 Comparison of the attractant effect of a mixture of isoamyl acetate and methyl eugenol and methyl eugenol on Bactrocera dorsalis

[0035] The experiment used a Y-arm olfactometer to determine and compare the selection tendency of Bactrocera dorsalis to the mixture and methyl eugenol. The specific experimental method was the same as in Example 1.

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

[0037] Example 3 Envelopment Experiment

[0038] The test was to compare the attracting effects of four treatments on the fruit fly, namely, a mixture of 50 μL / ml isoamyl acetate and 50% methyl eugenol, 50 μL / ml isoamyl acetate, 50% methyl eugenol, and paraffin oil. The blank attractant core was soaked in the four liquids for 2 hours, and then the different attractants were fixed in the center of the white sticky insect board. Then the four white sticky insect boards were randomly hung vertically on the top of the cage (1m×1m×1m). The room temperature was 28℃, the relative humidity was 65±5%, and ventilation was maintained. The test insects were immature female adults, sexually mature unmated female adults, and sexually mature mated female adults. 100 insects were put in each time, 50 males and 50 females. A small amount of water and feed was placed in the center of the floor of the cage to ensure the survival of the fruit flies. The test time period was selected from 12:00-15:00 every day. The number of attracted male and female insects on the insect attractant board was recorded with a camera every 5 minutes, and three groups were repeated.

[0039] The results showed that the mixture (i.e., 50 μL / ml of isoamyl acetate and 50% methyl eugenol) had the fastest attracting rate for all stages of the adult fruit fly (see 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 had a significant improvement in attracting adults of all stages (see Figure 4 ).

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

Claims

1. A bisexual attractant for Bactrocera dorsalis, characterized in that: The oriental fruit fly bisexual attractant comprises methyl eugenol and isoamyl acetate.

2. The two-sex attractant for the oriental fruit fly according to claim 1, comprising 40-60% v / v methyl eugenol and 40-60 μL / mL isoamyl acetate.

3. The bisexual attractant for the citrus fruit fly according to claim 2, which is a paraffin oil solution, wherein the active ingredients are 50% v / v methyl eugenol and 50 μL / mL isoamyl acetate.

4. The method for preparing the bisexual attractant of Bactrocera dorsalis according to claim 1, characterized in that: The method comprises: taking 900 μL of paraffin oil and 100 μL of isoamyl acetate and mixing them to obtain a 100 μL / ml isoamyl acetate solution; taking 500 μL of the 100 μL / ml isoamyl acetate solution and mixing it with 500 μL of methyl eugenol to obtain a mixed attractant containing 50 μL / ml isoamyl acetate and 50% methyl eugenol.

5. Use of the bisexual attractant for the citrus fruit fly according to any one of claims 1 to 3 in the preparation of pesticides or devices for trapping or killing the citrus fruit fly.

6. The use according to claim 5, wherein the pesticide or device for trapping or killing the oriental fruit fly is a pesticide or device for trapping or killing mated oriental fruit flies.

7. The use according to claim 6, wherein the pesticide or device for trapping or killing the citrus fruit fly is a pesticide or device for trapping or killing mated female citrus fruit flies.

8. The use according to claim 5, wherein the pesticide or device for trapping or killing the fruit fly is a pesticide or device for trapping or killing sexually immature female fruit flies.

9. The use according to any one of claims 5 to 8, wherein the device is a trap, a lure core or a sticky insect board.

10. A device for trapping or killing the fruit fly, comprising the fruit fly bisexual attractant according to any one of claims 1 to 3.

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