Plant volatile matter-based drosophila okadaiensis imago attractant and application thereof

Abstract

The invention discloses a drosophila okadaiensis imago attractant based on plant volatile matters and application thereof, and belongs to the field of insect control, the drosophila okadaiensis imago attractant comprises a female imago attractant: 0.2% of ethyl butyrate, 20% of n-hexyl alcohol and 20% of (2R, 3R)-(-)-2, 3-butanediol are mixed according to the volume ratio of 1: 1: 1; the male adult attractant is formed by mixing 20% of n-hexyl alcohol, 20% of (2R, 3R)-(-)-2, 3-butanediol and 0.2% of isoamyl alcohol according to the volume ratio of 1: 1: 1; the solvent is liquid paraffin. The botanical attractant which has high efficiency and species specificity for the vector and agricultural pest okadaioma drosophila is invented for the first time, and compared with a traditional attractant, the botanical attractant has the advantages that the trapping rate is increased, the botanical attractant has certain species specificity, and the botanical attractant has specific attraction capacity for sex. The method not only can be applied to pest control of the species in the field, but also has positive significance on dynamic monitoring and distribution of the vector insects, has an important effect on predicting the conjunctival sucking nematode disease propagated by the vector insects, and has a better application prospect.

Description

Technical Field

[0001] The invention relates to an insect trapping and control technology, in particular to an attractant for adult Okada fruit flies based on plant volatiles and an application thereof. Background Art

[0002] Okada's eyed fruit fly ( Phorticaokadai, P. okadai ) belongs to the subfamily Steganinae, genus Drosophila ( Phortica ) is an omnivorous fruit and vegetable pest. Its life cycle includes four stages: egg, larva, pupa and adult. Under the conditions of temperature 28 ℃, relative humidity 75% and light intensity 8 h / d, one generation can be completed in 25 days, and the generation cycle is relatively short. P. okadai It has a varied diet and can feed on more than 20 kinds of fruits and vegetables, including apples, pears, bananas, tomatoes, etc. The larvae penetrate into the flesh of the fruit to feed, causing damage and affecting the quality of the fruits and vegetables. More importantly, P. okadai They can also feed on the eye secretions of mammals, also known as eye-swallowing or tear-eating behavior (Lachryphagy). P. okadai When feeding on the eye secretions of mammals, the zoonotic parasitic nematode, the conjunctival sucking nematode ( Thelaziacallipaeda , T. callipaeda ) infective larvae spread to the eyes of the definitive host, causing T. callipaeda The transmission of the conjunctival sucking nematodes causes thelaziosis. The conjunctival sucking nematodes often occurs in rural areas or communities with relatively low living conditions and socioeconomic levels. Since the worms can be removed surgically or treated with antiparasitic drugs such as imibemycin oxime, it has not attracted enough attention from more clinicians and scholars and is a neglected parasitic disease. However, due to T. callipaeda There are many types of definitive hosts that can be parasitized, and the intermediate hosts are widely distributed. T. callipaeda Repeated infection of humans and animals is more serious.

[0003] Conjunctival sucking nematode disease can cause serious eye diseases in the later stages. At present, in addition to clinical surgery and killing adult dogs, there is still a lack of preventive drugs such as vaccines for this parasitic disease. Combined with the epidemiological characteristics of conjunctival sucking nematode disease, cutting off its transmission routes and conducting vector control are the only realistic and effective prevention and control strategies. At present, the control of vector insects is mainly carried out through chemical control, environmental control, biological control and other methods. Among them, chemical control is widely used because of its good effect and simple method. Chemical insecticides are an important means of chemical control. Because combined with anti-inflammatory drugs and other drug treatments, conjunctival sucking nematodes in the eyes can be effectively eradicated. For many years, conjunctival sucking nematode disease and vector insect control have not received enough attention. Therefore, at present, for P. okadai No special measures have been taken for prevention and control, and the pests are mainly killed by spraying pesticides in agricultural production.

[0004] Currently, due to the large-scale, high-dose and high-frequency use of chemical pesticides, new insect-resistant strains are constantly being reported after long-term natural selection. The current problem of insecticide resistance is extremely serious. According to statistics, more than 600 species of insects have been found to have resistance, and the resistance level of existing resistant insects is still increasing. P. okadai Due to their feeding needs, they are mainly distributed around orchards. However, in actual agricultural production, pesticides are often used in large doses and at high frequencies, especially pyrethroid insecticides, which can cause P. okadai The chances of receiving the drug increase, and P. okadai and Drosophila melanogaster ( Drosophila melanogaster , D . melanogaster ), have the characteristics of strong reproductive ability, short generation cycle, high inbreeding rate, etc., which makes P. okadai It is easy to develop resistance to pesticides. The applicant has caught P. okadai We speculate that after long-term natural selection pressure, it has developed a certain resistance to insecticides, and even strains in some areas may have developed a certain resistance, which will increase the risk of transmission of conjunctival sucking nematodes.

[0005] P. okadai It is more active in an environment with a temperature of 20-25 ℃ and a humidity of 50%-70%. It is distributed in many areas such as Guizhou, Anhui, Hebei, Henan, Hubei and Sichuan. In recent years, there have been continuous cases of conjunctival sucking nematode disease in Guizhou, and the actual number of infections may be much higher than the reported number. P. okadai Research on the callipaeda species has primarily focused on morphology, with research on its ecological niche and attractants lagging behind. Urgently conducting ecological research on biological control, particularly attractant development, is crucial to effectively curbing the spread of T. callipaeda. Summary of the Invention

[0006] The purpose of the present invention is to overcome the above-mentioned background technical difficulties and provide an Okada's eye-catching fruit fly adult attractant based on plant volatiles and its application.

[0007] In order to achieve the above purpose, the technical solutions adopted are: A plant volatile-based attractant for Drosophila okadaii adults comprises a female adult attractant and a male adult attractant, wherein the female adult attractant is a mixture of 0.2% ethyl butyrate, 20% n-hexanol, and 20% (2R,3R)-(-)-2,3-butanediol in a volume ratio of 1:1:1; the male adult attractant is a mixture of 20% n-hexanol, 20% (2R,3R)-(-)-2,3-butanediol, and 0.2% isoamyl alcohol in a volume ratio of 1:1:1; and the solvent is liquid paraffin.

[0008] And the use of the above-mentioned plant volatile-based Okada's eye-catching fruit fly adult attractant for trapping Okada's eye-catching fruit fly adults.

[0009] Furthermore, the attractants are all carried in a barrel-shaped plastic trap with holes and a lid; the prepared attractant is added to a concave rubber head to form a lure core, and the total amount of prepared plant volatiles and liquid paraffin corrosion inhibitor added to each rubber head is 120-150 μl, and the lure core is hung in the barrel-shaped trap; then the device is placed in an orchard to trap Okada's eyed fruit fly.

[0010] The beneficial effects of adopting the above scheme include the invention of a highly effective and species-specific plant-based attractant for the disease vector and agricultural pest Drosophila okadaiana. Compared with traditional attractants composed of red wine and apple cider vinegar or fermented juice, this attractant improves the Drosophila okadaiana's ability to attract flies. It also exhibits a certain degree of species specificity and has a specific ability to attract both sexes. This invention is not only applicable to pest control of this species in the field, but also importantly has positive implications for the dynamic monitoring and distribution of this disease-carrying insect, playing an important role in predicting the conjunctival sucking nematode disease it transmits, and has promising application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 Female and male after mating for different fruit pairs P.okadai attractiveness; Figure 2 For different fruit pairs, unmated females and males P.okadai attractiveness; Figure 3 This is the GC-MS peak diagram of apple volatiles; Figure 4 This is the GC-MS peak diagram of pear volatiles; Figure 5 This is the GC-MS peak diagram of banana volatiles; Figure 6 These are the common and differential volatile components of the three fruits before and after fermentation; Figure 7 The contents of the common volatile components in bananas before and after fermentation; Figure 8 is the content of common differential volatile components in apples before and after fermentation; Figure 9 is the content of common differential volatile components in pears before and after fermentation; Figure 10 For the preference of fruit volatile components for females P. okadai attractiveness; Figure 11 For the preference of fruit volatile components for male P. okadai attractiveness; Figure 12 For different attractants P. okadai Comparison of the lure effect; Figure 13 As an attractant, fermented pear juice for 3 days P. okadai Comparison of attractiveness; Figure 14 As an attractant, fermented pear juice for 3 days D. melanogaster Comparison of attractiveness. DETAILED DESCRIPTION

[0012] The technical solutions of the present invention are described clearly and completely below in conjunction with specific embodiments of the present invention. The described embodiments are only a portion of the embodiments of the present invention, not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative work are within the scope of protection of the present invention.

[0013] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

[0014] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

[0015] 1 Plant volatile-based attractant for Okada's Drosophila adult flies The invention comprises a female adult attractant and a male adult attractant, wherein the female adult attractant is a mixture of 0.2% ethyl butyrate, 20% n-hexanol, and 20% (2R, 3R)-(-)-2,3-butanediol in a volume ratio of 1:1:1; and the male adult attractant is a mixture of 20% n-hexanol, 20% (2R, 3R)-(-)-2,3-butanediol, and 0.2% isopentanol in a volume ratio of 1:1:1.

[0016] 2 Experimental materials and equipment 2.1 Experimental Materials (1) Insect breeding and handling Research use P . okadaiWe collected it in Zunyi City (27 13′ 15″ N, 106 17′ 22″E) in 2017 and then cultured it in the Pathogenic Biology Laboratory of Zunyi Medical University. P . okadai The adults were fed with fermented apples and pears for three days, and were kept in cages with nylon mesh on all sides. The temperature was 28 ± 2°C, the relative humidity was 75 ± 10%, and the light / dark cycle was 12:12. P . okadai For 7-day-old adult flies, they were starved for 12 hours before the experiment.

[0017] (2) Different fruits and their processing The fruits and vegetables used in the experiment were all fresh fruits purchased from supermarkets. The fermentation conditions were a constant temperature box in the natural environment of the laboratory. Before fermentation, the fruits were peeled and cut into 2*2*2cm pieces. The fermentation temperature was 28℃.

[0018] 2.2 Experimental Reagents Table 1 Main experimental reagents Brand Name Reagent name CAS number Specification English Macklin's reagent liquid paraffin 8042-47-5 AR 99% Paraffin liquid Macklin's reagent Anhydrous ethanol 64-17-5 99.50% Ethanol Macklin's reagent Acetic acid 64-19-7 ≥99.8%(GC) Acetic acid Macklin's reagent Isoamyl alcohol 123-51-3 ≥99.5% (GC) 1-Butanol, 3-methyl- Macklin's reagent Ethyl butyrate 105-54-4 ≥99.5% (GC) Butanoic acid, ethyl ester Macklin's reagent n-Hexanol 111-27-3 ≥99.5% (GC) 1-Hexanol Macklin's reagent Ethyl acetate 141-78-6 ≥99.7% (GC) Ethyl acetate Macklin's reagent 2-Phenylethanol 1960-12-8 ≥99.5% (GC) Phenylethyl Alcohol Macklin's reagent 2-Octanone 111-13-7 ≥99.5% (GC) 2-Octanone Macklin's reagent Acetoin 513-86-0 97% Acetoin Macklin's reagent Isoamyl acetate 123-92-2 ≥99.5% (GC) 1-Butanol, 3-methyl-, acetate Macklin's reagent (2R,3R)-(-)-2,3-Butanediol 24347-58-8 97% 2,3-Butanediol, [R-(R*,R*)]- Macklin's reagent 2-octanol 123-96-6 ≥99.5% (GC) 2-Octanol 2.3 Experimental Instruments Table 2 Main experimental instruments Brand Name Instrument name or model self made Y-type olfactometer self made Four-arm olfactometer Shanghai Boxun Biological Co., Ltd. Artificial climate incubator (BIC-400 model) snowflake Ice Maker Ike water purifier Agilent 7890B-5977B Leica Gas Chromatography-Mass Spectrometry Stereo Microscope Sf9i

[0019] 3 Experimental methods 3.1 Determination of the attractiveness of different fruits to Drosophila okadaensis Eight fruits, including pear, apple, banana, strawberry, blueberry, pitaya, tangerine, and grape, were peeled and naturally fermented for 1-3 days. Fresh, unfermented fruits served as controls. Behavioral experiments were conducted using a four-arm olfactometer. Each experiment involved 20 unmated or mated female or male fruit flies, with 20 replicates for each fruit type. During the experiment, fruits fermented for different days were placed in four attractant tubes of the four-arm olfactometer, with a fruit fly starved for 12 hours placed in the center. The entire experiment was conducted in a light-free incubator at a constant temperature (28°C) and humidity (80%). Results were recorded after 12 hours.

[0020] 3.2 Analysis of volatile components in fruits preferred by Drosophila okadaensis (1) Preference for fruit volatile component detection Apples, pears, and bananas, which are highly attractive to Drosophila okadaii, were selected. Three-day fermented juice was used as treatment, and fresh juice was used as control. Equal weights of juice were taken from each sample (Table 3). Solid-phase microextraction (SPE) was used with 2-octanol as the internal standard, and the volatiles of the juice were analyzed by gas chromatography-mass spectrometry (GC-MS). Each sample was repeated five times. Specific conditions were as follows: chromatographic column: DB-wax (30 m × 0.25 mm × 0.25 µm); liner: splitless, without glass wool; extraction head: 50 / 30 μm, DVB / CAR / PDMS; injection temperature: 260°C; carrier gas: helium; flow rate: 1 mL / min; column temperature: 40°C (hold for 5 min), ramped to 220°C at 5°C / min, then to 250°C at 20°C / min, held for 2.5 min; ion source temperature: 230°C; quadrupole temperature: 150°C; scan mode: full scan; mass range: 20–400; desorption time: 5 min; GC cycle time: 50 min. Chromatographic-mass spectrometric analysis was performed using Shimadzu GCMSsolution 4.42 software, and spectral matching was performed using the Shimadzu NIST14 library.

[0021] Table 3 Juice weight pear Weight (g) apple Weight (g) banana Weight (g) P3-1 3.293 A3-1 3.5 B3-1 3.5 P3-2 3.295 A3-2 3.5 B3-2 3.5 P3-3 3.296 A3-3 3.5 B3-3 3.5 P3-4 3.294 A3-4 3.5 B3-4 3.5 P3-5 3.296 A3-5 3.5 B3-5 3.5 P0-1 3.295 A0-1 3.5 B0-1 3.5 P0-2 3.284 A0-2 3.5 B0-2 3.5 P0-3 3.295 A0-3 3.5 B0-3 3.5 P0-4 3.293 A0-4 3.5 B0-4 3.5 P0-5 3.301 A0-5 3.5 B0-5 3.5 (2) Analysis of differential volatile components Taking 2-octanol as the internal standard substance, the peak area ratio of all substances was taken to calculate the relative content of each component. The relative content of the same substance was calculated using the difference method. The components with a larger increase compared with the fresh juice with poor attraction effect were selected as candidate components. The comprehensive results of the three fruits were combined to determine the final possible attraction. P . okadai ingredients.

[0022] 3.3 Determination of the attractiveness of Drosophila okadaensis to the volatile components of preferred fruits A mixture of 7.2% ethanol and 1.6% acetic acid was used as solvent to prepare solutions of different concentrations of each volatile component, with a high concentration of 20%, a medium concentration of 2%, and a low concentration of 0.2%. P . okadai Behavioral measurements were performed using a four-arm olfactometer.

[0023] 3.4 Screening of plant-derived attractants Using 7.2% ethanol + 1.6% acetic acid as solvent, we designed different attractant compositions by using the effective concentration of volatile components with attractive ability, and used a four-arm olfactometer and a Y-type olfactometer to conduct behavioral tests in the laboratory to screen for effective attractant formulas. At the same time, using pear juice fermented for 3 days as a control, we used a Y-type olfactometer to verify the effects of the selected attractants on the attractant. P . okadai and D . melanogaster ability to attract.

[0024] 3.6 Data Analysis The results of the behavioral luring experiment were first analyzed using the Shapiro-Wilk test for normality and the Brown-Forsythe test for homogeneity of variance in the GraphPad Prism 9 software. t Unpaired test or Welch correction t Significant differences between experimental groups were assessed using a one-way analysis of variance or the nonparametric Kruskal-Wallis test. If the data did not conform to a normal distribution, the Brown-Forsythe test and the Welch test were used. * indicates a significant difference at the 0.05 level, and ** indicates a significant difference at the 0.01 level.

[0025] 4 Experimental Results 4.1 Attractiveness of different fruits to Drosophila okadaensis Among the eight fruits selected, namely pear, apple, banana, strawberry, blueberry, orange, dragon fruit and grape, P . okadai The preference for fermented fruits is stronger than that for unfermented fruits. Specifically, the preference for fermented pears, apples, strawberries and bananas for 3 days is stronger for both mated and unmated males and females. P . okadai The appeal is stronger than fresh or fermented 1-2 days of blueberries, oranges, dragon fruit and grapes ( Figure 1 ah, Figure 2 ah); all treatments for blueberry and dragon fruit P . okadai There is no difference in attractiveness ( Figure 1 il, Figure 2 il); fermented grapes for 3 days after mating, female and male P. okadai was more attractive than the other treatment groups, but was less attractive to unmated females. P. okadai Fresh grapes showed the most significant difference in attractiveness to unmated males. P. okadaiThere was no difference in attractiveness among the treatment groups ( Figure 1 m and n, Figure 2 m and n); oranges fermented for 1 day showed significant differences in attractiveness to mated males and unmated females, but no significant differences in attractiveness to mated females ( Figure 1 o and p, Figure 2 o and p).

[0026] Figure 1 Female and male after mating for different fruit pairs P. okadai a and b, different treatments of pear P. okadai c and d, different treatments of apples P. okadai e and f, different treatments of bananas P. okadai g and h, different treatments of strawberries P. okadai i and j, different treatments of blueberries P. okadai k and l, different treatments of pitaya on P. okadai m and n, different treatments of grapes P. okadai o and p, different treatments of oranges on P. okadai 0 means fresh; 1 means fermented for 1 day; 2 means fermented for 2 days; 3 means fermented for 3 days. * means p <0.05; ** indicates p <0.01.

[0027] Figure 2 For different fruit pairs, unmated females and males P. okadai Among them, a and b, different treatments of pear P. okadai c and d, different treatments of apples P. okadai e and f, different treatments of bananas P. okadai g and h, different treatments of strawberries P. okadai i and j, different treatments of blueberries P. okadai k and l, different treatments of pitaya on P. okadai m and n, different treatments of grapes P. okadai o and p, different treatments of oranges on P. okadai 0 means fresh; 1 means fermented for 1 day; 2 means fermented for 2 days; 3 means fermented for 3 days. * means p <0.05; ** indicates p <0.01; *** indicates p <0.001.

[0028] 4.2 Analysis of common volatile components in fruits preferred by Drosophila okadaii Pear, apple and banana were selected as the fruits preferred by Drosophila. Through volatile metabolomics analysis, it was found that there were 66 volatile compounds in pear (see Figure 3 ), apple has a total of 70 volatile compounds ( Figure 4 ), banana has a total of 129 volatile compounds ( Figure 5 Compared with fresh pear juice, the contents of 63 volatile components increased in pear juice after 3 days of fermentation. Compared with fresh apple juice, the contents of 65 components increased in apple juice after 3 days of fermentation. Compared with fresh banana juice, the contents of 99 components increased in banana juice after 3 days of fermentation.

[0029] Figure 3 GC-MS peak profiles of apple volatiles. a, blank control. b, fresh apple juice. c, three-day fermented apple juice. The arrow indicates the internal standard, 2-octanol, with a retention time of 17.5.

[0030] Figure 4 GC-MS peak profiles of pear volatiles. a, blank control. b, fresh pear juice. c, 3-day fermented pear juice. The arrow indicates the internal standard 2-octanol, with a retention time of 17.5.

[0031] Figure 5 GC-MS peak profiles of banana volatiles. a, blank control. b, fresh banana juice. c, three-day fermented banana juice. The arrow indicates the internal standard, 2-octanol, with a retention time of 17.5.

[0032] The results of analyzing the different volatile components of the three fruits before and after fermentation showed that among these different components, a total of 23 components were present in the three fruits ( Figure 6 ), further screening components with large content differences, a total of 11 volatile components ethanol (Ethanol), ethyl acetate (Ethyl acetate), (2R,3R)-(-)-2,3-butanediol (2,3-Butanediol, [R-(R*,R*)]-), acetic acid (Acetic acid), 2-phenylethanol (Phenylethyl alcohol), butanoic acid (Butanoicacid, ethyl ester), isopentanol (1-Butanol, 3-methyl-), isopentanol (1-Butanol, 3-methyl-, acetate), n-hexanol (1-Hexanol), acetoin and 2-octanone (2-Octanone) were selected as candidate small molecules, and their expression levels in various fruits are shown in the following table. Figure 7 、 8and 9.

[0033] Figure 6 Common and differential volatile components of three fruits before and after fermentation.

[0034] Figure 7 The contents of common volatile components in bananas before and after fermentation. B0: fresh banana; B3: banana fermented for three days.

[0035] Figure 8 The contents of common volatile components in apples before and after fermentation. A0: fresh apples; A3: apples fermented for three days.

[0036] Figure 9 The contents of common volatile components in pears before and after fermentation. P0: fresh pears; P3: pears fermented for three days.

[0037] 4.3 Determination of the attractiveness of preferred fruit volatiles to Drosophila okadaensis Behavioral measurements were performed using a four-arm olfactometer using a mixture of 7.2% ethanol and 1.6% acetic acid as solvents and blank controls. The results showed that 20% n-hexanol and 20% (2R,3R)-(-)-2,3-butanediol were attractive to both male and female Drosophila okadaensis, 0.2% isoamyl alcohol was attractive to males, and 0.2% ethyl butyrate and 20% acetoin were attractive to females. Figure 10 , Figure 11 ).

[0038] Figure 10 For the preference of fruit volatile components for females P. okadai The attraction of the air. ai, respectively, represents the volatile components of different concentrations, where a, 2-phenylethanol; b, 2-octanone; c, acetoin; d, ethyl acetate; e, isoamyl acetate; f, (2R,3R)-(-)-2,3-butanediol; g, ethyl butyrate; h, isoamyl alcohol; i, n-hexanol. * indicates p <0.05; ** indicates p <0.01.

[0039] Figure 11 Preferred fruit volatiles for males P. okadai The attraction of the air. ai, respectively, represents the volatile components of different concentrations, where a, 2-phenylethanol; b, 2-octanone; c, acetoin; d, ethyl acetate; e, isoamyl acetate; f, (2R,3R)-(-)-2,3-butanediol; g, ethyl butyrate; h, isoamyl alcohol; i, n-hexanol. * indicates p <0.05; ** indicates p <0.01.

[0040] 4.4 Screening of plant-derived attractants for Drosophila okadaii (1) P. okadai Screening of plant-derived attractants Using 7.2% ethanol + 1.6% acetic acid as solvent, the effective concentration of volatile components with attractive ability was used to conduct orthogonal design using L4 (2^4) orthogonal array method, targeting females and males respectively. P. okadai The composition of different attractants was designed (Table 4, Table 5). The behavioral experimental results showed that 0.2% ethyl butyrate, 20% n-hexanol, 20% (2R,3R)-(-)-2,3-butanediol mixed with 7.2% ethanol and 1.6% acetic acid had a negative effect on female P. okadai The most attractive Figure 12 ). 20% n-hexanol, 20% (2R,3R)-(-)-2,3-butanediol, 0.2% isopentanol, 7.2% ethanol and 1.6% acetic acid were mixed and used to treat male P. okadai The most attractive Figure 12 ).

[0041] Table 4 Female P. okadai Plant-derived attractant design Note: 1 indicates the effective concentration of this ingredient, and 2 indicates that the concentration of this ingredient is 0 Test No. 0.2% ethyl butyrate 20% n-hexanol 20% (2R,3R)-(-)-2,3-Butanediol 20% acetoin 1 2 1 2 2 2 1 2 2 1 3 2 1 2 1 4 2 2 1 1 5 1 2 2 2 6 1 1 1 1 7 2 2 1 2 8 1 1 1 2 Table 5 Male P. okadai Plant-derived attractant design Note: 1 indicates the effective concentration of this ingredient, and 2 indicates that the concentration of this ingredient is 0 Test No. 20% n-hexanol 20% (2R,3R)-(-)-2,3-Butanediol 0.2% isoamyl alcohol A1 1 1 1 B2 1 2 1 C3 1 2 2 D4 2 2 1 Table 5 Male P. okadai Design of plant-derived attractants (continued) Note: 1 indicates the effective concentration of this ingredient, and 2 indicates that the concentration of this ingredient is 0 Test No. 20% n-hexanol 20% (2R,3R)-(-)-2,3-Butanediol 0.2% isoamyl alcohol E5 1 1 2 F6 2 1 2 J7 2 2 2 H8 2 1 1 Figure 1 2 for different attractants P. okadai Comparison of the attractant effects of different small molecule combinations and different types of attractants on females. a, b, c are the effects of different small molecule combinations and different types of attractants on females. P. okadai Comparison of the number of attractants; d, e, f are the effects of different small molecule combinations of different types of attractants on males. P. okadai Comparison of the number of attractants. Different letters indicate significant differences at the 0.05 level, and the same letters indicate insignificant differences.

[0042] (2) P. okadai Attraction of plant-derived attractants to Drosophila melanogaster Furthermore, the attractants screened above were used, and pear juice was used as a control to determine the effects of different small molecule mixtures on the P. okadaiand Drosophila melanogaster, the results showed that: female P. okadai Attractants No. 6 and No. 8, P. okadai The attraction of attractant No. 6 to female Drosophila melanogaster was also significantly stronger than that of pear juice fermented for 3 days ( Figure 13 , Figure 14 ).male P. okadai A1 attractant, P. okadai The attractiveness of the two juices was significantly stronger than that of pear juice fermented for 3 days, but the attractiveness to male Drosophila melanogaster was not significant compared with that of pear juice fermented for 3 days.

[0043] Figure 13 Effect of attractant and 3-day fermented pear juice P. okadai Comparison of attractiveness of different combinations of attractants and fermented 3-day pear juice on females. a, b are the effects of different combinations of attractants and fermented 3-day pear juice on females. P. okadai Comparison of the number of attractants; c, d are the effects of different combinations of attractants and fermented 3-day pear juice on male P. okadai Comparison of the number of lures. * indicates p <0.05; ** indicates p <0.01.

[0044] Figure 14 Effect of attractant and 3-day fermented pear juice D. melanogaster Comparison of attractiveness of different combinations of attractants and fermented 3-day pear juice on females. D. melanogaster Comparison of the number of attractants; b is the effect of different combinations of attractants and fermented 3-day pear juice on male D. melanogaster ** indicates the number of lures. p <0.01.

Claims

1. A plant volatile-based attractant for Drosophila okadai, characterized by: The invention comprises a female adult attractant and a male adult attractant, wherein the female adult attractant is a mixture of 0.2% ethyl butyrate, 20% n-hexanol, and 20% (2R,3R)-(-)-2,3-butanediol in a volume ratio of 1:1:1; the male adult attractant is a mixture of 20% n-hexanol, 20% (2R,3R)-(-)-2,3-butanediol, and 0.2% isoamyl alcohol in a volume ratio of 1:1:1; and the solvent is liquid paraffin.

2. Use of the plant volatile-based Drosophila okadaensis adult attractant according to claim 1 for trapping Drosophila okadaensis adults.

3. The use according to claim 2, characterized in that: The attractants are all carried in a barrel-shaped plastic trap with holes and a lid; the prepared attractants are added to a concave rubber head to form a lure core, and the total amount of prepared plant volatiles and liquid paraffin corrosion inhibitor added to each rubber head is 120-150 μl, and the lure core is hung in the barrel-shaped trap; then the device is placed in an orchard to trap Okada's eyed fruit flies.

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