A kind of scarlet hair moth attractant composition and application

Abstract

The present invention discloses a vermilion hairy moth attractant composition and application. The active substances of the composition are composed of 1 to 10 μg / μL of cis-9-hexadecenal, 10 to 100 μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal, 10 to 100 μg / μL of (-)-α-pinene, 10 to 100 μg / μL of β-myrcene, 10 to 100 μg / μL of β-ocimene, and 1 to 100 μg / μL of β-caryophyllene. The active substances can be further obtained by using paraffin as a solvent to obtain an attractant. The attractant has a significant attracting effect, is used in a small amount, is easy to use, and has a strong target property. The attractant can be simultaneously applied to the vermilion hairy moth prevention and control and population dynamics monitoring, and is of great significance for the research and development of new technologies for the green prevention and control of the vermilion hairy moth.

Description

Technical Field

[0001] The invention belongs to the technical field of biological control, and particularly relates to an attractant composition for the red hairy moth and an application thereof. Background Art

[0002] The scarlet hairy moth, Phauda flammans (Walker), belongs to the family Phaudidae of the order Lepidoptera. It primarily harms plants in the genus Ficus, with Ficus benjamina and F. microcarpa being the most severely affected. In recent years, outbreaks have occurred in southern China. After hatching, the larvae of the scarlet hairy moth feed from the treetops downwards, resulting in the affected plants often exhibiting a "bald" appearance. In severe cases, the entire tree is left with only a bare trunk, severely impacting not only the growth and development of the banyan tree but also the urban landscape and ecological benefits. This pest has a long and explosive period of occurrence and damage, and is currently primarily controlled by chemical means. However, chemical control can easily cause environmental pollution and impact human health. Therefore, there is an urgent need to develop green and healthy control technologies. Summary of the Invention

[0003] The purpose of the present invention is to provide a red hairy moth attractant composition and application, so as to achieve the effect of green and healthy prevention and control of red hairy moth.

[0004] To achieve the above objectives, the present invention provides the following technical solutions:

[0005] A red hairspot moth attractant composition, the active substances of which are composed of sex pheromones and plant volatiles, wherein the sex pheromones are cis-9-hexadecenal and (Z,Z,Z)-9,12,15-octadecatrienal, and the plant volatiles are (-)-α-pinene, β-myrcene, β-ocimene and β-caryophyllene.

[0006] The following scheme is further preferred:

[0007] A red hairspot moth attractant composition, the active substances of which are composed of 1-10 μg / μL cis-9-hexadecenal, 10-100 μg / μL (Z,Z,Z)-9,12,15-octadecatrienal, 10-100 μg / μL (-)-α-pinene, 10-μg / μL β-myrcene, 10-100 μg / μL β-ocimene, and 1-100 μg / μL β-caryophyllene.

[0008] The following scheme is further preferred:

[0009] A red hairspot moth attractant composition, the active ingredients of which are composed of 1 μg / μL of cis-9-hexadecenal, 10 μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal, 100 μg / μL of (-)-α-pinene, 10 μg / μL of β-myrcene, 10 μg / μL of β-ocimene and 1 μg / μL of β-caryophyllene.

[0010] Alternatively, its active ingredients consist of 1 μg / μL of cis-9-hexadecenal, 10 μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal, 100 μg / μL of (-)-α-pinene, 10 μg / μL of β-myrcene, 100 μg / μL of β-ocimene and 100 μg / μL of β-caryophyllene.

[0011] Alternatively, its active ingredients consist of 10 μg / μL of cis-9-hexadecenal, 100 μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal, 100 μg / μL of (-)-α-pinene, 10 μg / μL of β-myrcene, 10 μg / μL of β-ocimene and 1 μg / μL of β-caryophyllene.

[0012] The present invention also claims the use of the aforementioned attractant composition to prepare an attractant for the red hairy moth. The attractant is prepared by diluting the sex pheromone and plant volatiles with liquid paraffin to a certain concentration, adding a certain volume of the mixture to a lure according to the dosage, and storing the lure in a refrigerator at -20°C for future use.

[0013] The present invention also claims to protect the use of the above-mentioned composition or attractant in the control and population monitoring of the red hair moth.

[0014] The present invention has the following beneficial effects:

[0015] The invention has good synergistic effects, has a strong trapping effect on male vermilion hairy moths, is environmentally friendly, harmless to humans, can be accurately applied to the prediction and trapping of vermilion hairy moths, and has good economic and ecological benefits. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Figure 2 shows the antennal electrophysiological responses of male Trichoderma vermilionii to different concentrations of cis-9-hexadecenal and (Z,Z,Z)-9,12,15-octadecatrienal. A corresponds to cis-9-hexadecenal; B corresponds to (Z,Z,Z)-9,12,15-octadecatrienal. Data are mean ± standard error. Different lowercase letters above the bars indicate significant differences between treatments after Tukey's HSD multiple comparison (P < 0.05).

[0017] Figure 2 Figure 2 shows the directional behavioral responses of male Trichoderma vermilionii to different concentrations of cis-9-hexadecenal and (Z,Z,Z)-9,12,15-octadecatrienal. A corresponds to cis-9-hexadecenal; B corresponds to (Z,Z,Z)-9,12,15-octadecatrienal. ** and * indicate significant differences between the treatment and the control at P < 0.01 and P < 0.05, respectively; NS indicates no significant difference between the treatment and the control at P < 0.05 (independent sample t-test).

[0018] Figure 3 Figure 2 shows the antennal electrophysiological responses of male Trichoderma vermilion to six different concentrations of two sex pheromones. A through F represent mixtures of cis-9-hexadecenal and (Z,Z,Z)-9,12,15-octadecatrienal at concentrations of 1 + 10, 1 + 100, 10 + 10, 10 + 10, 100 + 10, and 100 + 100 μg / μL, respectively. Data are mean ± standard error; different lowercase letters within error bars indicate significant differences at the P < 0.05 level (Tukey's HSD test).

[0019] Figure 4 This figure shows the directional behavioral responses of male tricholoma cinnamomi to the optimal combination of two sex pheromones. A and D represent mixtures of cis-9-hexadecenal and (Z,Z,Z)-9,12,15-octadecatrienal at concentrations of 1 + 10 and 10 μg / μL + 100 μg / μL, respectively; the control was liquid paraffin. Data are mean ± standard error; NR indicates individuals who did not respond to treatment; N indicates total sample size; * indicates a significant difference between treatment and control at P < 0.05 (independent sample t-test).

[0020] Figure 5 Figure 2 shows the antennal electrophysiological responses of male Trichoderma vermilionii to different concentrations of (-)-α-pinene, β-myrcene, β-ocimene, and β-caryophyllene. A, (-)-α-pinene; B, β-myrcene; C, β-ocimene; D, β-caryophyllene. Data are mean ± standard error. Different lowercase letters within error bars indicate significant differences at the 0.05 level (Tukey's HSD test).

[0021] Figure 6Figure 2. Orienting behavioral responses of male Trichoderma vermilionii to different concentrations of (-)-α-pinene, β-myrcene, β-ocimene, and β-caryophyllene. A, (-)-α-pinene; B, β-myrcene; C, β-ocimene; D, β-caryophyllene. Data are mean ± standard error. NR indicates individuals that did not respond to treatment. N indicates total sample size. ** and * indicate significant differences between treatment and control at P < 0.01 and P < 0.05, respectively. NS indicates no significant difference between treatment and control at P < 0.05 (independent sample t-test).

[0022] Figure 7 The antennal electrophysiological responses of male Ficus microcarpa moths to eight different concentrations of four active ingredients from volatiles of Ficus microcarpa are shown in the present invention. G to N represent mixtures of (-)-α-pinene, β-myrcene, β-ocimene, and β-caryophyllene at concentrations of 100+10+10+1, 100+10+10+100, 100+10+100+1, 100+10+100+100, 100+100+10+1, 100+100+10+100, 100+100+100+1, and 100 μg / μL+100 μg / μL+100 μg / μL+100 μg / μL+100 μg / μL, respectively. Data are mean ± standard error; different lowercase letters within error bars indicate significant differences at the 0.05 level (Tukey's HSD).

[0023] Figure 8 This figure shows the directional behavioral response of male Ficus microcarpa moths to the optimal ratio of four active ingredients in volatiles from Ficus microcarpa. G, I, and J represent mixtures of (-)-α-pinene, β-myrcene, β-ocimene, and β-caryophyllene at concentrations of 100+10+10+1, 100+10+100+1, and 100μg / μL+10μg / μL+100μg / μL+100μg / μL, respectively. Data in the figure are mean ± standard error; NR indicates individuals who did not respond to the treatment; N indicates the total sample size; ** and * indicate significant differences between the treatment and the control at P < 0.01 and P < 0.05 (independent sample t-test).

[0024] Figure 9The antennal electrophysiological response of the male moth of the present invention to six mixtures of sex pheromones and volatile active components of Ficus microcarpa is shown. A+G, A+I, A+J, D+G, D+I and D+J represent mixtures of cis-9-hexadecenal, (Z,Z,Z)-9,12,15-octadecatrienal, (-)-α-pinene, β-myrcene, β-ocimene and β-caryophyllene at concentrations of 1+10+100+10+10+1, 1+10+100+10+100+1, 1+10+100+10+100+100, 10+100+100+10+10+1, 10+100+100+10+100+1 and 10 μg / μL+100 μg / μL+100 μg / μL+100 μg / μL+100 μg / μL+100 μg / μL, respectively. The data in the figure are mean ± standard error; different lowercase letters on the error bars indicate significant differences at the P < 0.05 level (Tukey's HSD test).

[0025] Figure 10 The present invention shows the directional behavioral responses of male tricholoma cinnamomi to three optimal ratios of sex pheromones and volatile active ingredients from Ficus microcarpa. A+G, A+J, and D+G represent the ratios of cis-9-hexadecenal, (Z,Z,Z)-9,12,15-octadecatrienal, (-)-α-pinene, β-myrcene, β-ocimene, and β-caryophyllene at concentrations of 1+10+100+10+10+1, 1+10+100+10+100+100, and 10+100 μg / μL+100 μg / μL+100 μg / μL+10 μg / μL+10 μg / μL+1 μg / μL, respectively; the control is liquid paraffin. The data in the figure are mean ± standard error; NR indicates individuals that did not respond to the treatment; N indicates the total sample size; ** and * indicate significant differences between the treatment and the control at P < 0.01 and P < 0.05, respectively (independent sample t test).

[0026] Figure 11The invention shows the attracting effect of the sex pheromone and the active ingredients of the volatile matter of Ficus microcarpa on the male insects of the red hair moth. A+G indicates that the concentration ratio of cis-9-hexadecenal, (Z,Z,Z)-9,12,15-octadecatrienal, (-)-α-pinene, β-myrcene, β-ocimene and β-caryophyllene is 1 μg / μL+10 μg / μL+100 μg / μL+10 μg / μL+10 μg / μL+1 μg / μL; the controls are liquid paraffin, A (1 μg / μL cis-9-hexadecenal+10 μg / μL (Z,Z,Z)-9,12,15-octadecatrienal) and G (100 μg / μL (-)-α-pinene+10 μg / μL β-myrcene+10 μg / μL β-ocimene+1 μg / μL β-caryophyllene); LP indicates liquid paraffin. The data in the figure are mean ± standard error; * indicates significant difference between treatment and control at P < 0.05 level (independent sample t test); different uppercase and lowercase letters indicate significant difference in selection rate between treatment group and control group by Tukey's HSD test (P < 0.05). DETAILED DESCRIPTION

[0027] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the accompanying drawings and specific embodiments. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0028] Example 1

[0029] Electrophysiological and directional behavioral responses of male Trichoderma vermilion to a single sex pheromone component

[0030] The single component of the sex pheromone of the scarlet hairy moth includes 0.01-100 μg / μL of cis-9-hexadecenal and 0.01-100 μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal.

[0031] The method for using the single-component sex pheromone of the scarlet hairy moth is to use liquid paraffin as a solvent to dissolve cis-9-hexadecenal and (Z,Z,Z)-9,12,15-octadecatrienal into a solution of 0.01 to 100 μg / μL.

[0032] 1.1 Antennae electrophysiological responses of male Trichoderma vermilion to single sex pheromone components

[0033] Using paraffin as a solvent, the two sex pheromone active substances were diluted into 5 concentration gradients of 0.01μg / μL, 0.1μg / μL, 1μg / μL, 10μg / μL and 100μg / μL, paraffin as a control and 0.1μg / μL leaf alcohol as a standard reference.

[0034] Before the test, male C. vermilionii moths were stunned with CO2. Surgical scissors were used to cut off the antennae from the base and the distal 1-2 segments were removed. The antennae were lifted with an insect pin and connected to metal electrodes coated with conductive glue at both ends. The antennae were placed approximately 1 cm from the outlet of the odor tube, with a continuous airflow of 300 mL / min. -1 After the antennal potential baseline stabilizes, the test begins. Each time, 5 μL of the test solution is aspirated and dropped onto a 1 cm × 2 cm qualitative filter paper. The solution is quickly placed into a 1 mL pipette tip and then connected to the stimulation tube. The stimulation airflow is set to 400 mL min -1 Each stimulation lasted 0.5 seconds, with a 60-second interval between stimulations to ensure that the antennal sensilla regained some activity. Using 0.1 μg / μL leaf alcohol as the standard reference, each test group was performed in the following order: control, standard reference, test sample, control, standard reference, and test sample. Only one antennae per male was used, and only one group was tested. Each sample was repeated five times.

[0035] The calculation formula for the relative value of antennal electrophysiological response is as follows:

[0036]

[0037] Statistical analysis: SPSS 26.0 software was used for data statistics. The EAG responses of the red-spotted moth to different concentrations of volatile standards of Ficus microcarpa were analyzed by one-way analysis of variance (ANOVA), and the Turkey method was used for significance test (P < 0.05).

[0038] The results are as follows Figure 1 As shown, the relative values ​​of antennal electrophysiological responses of males of the red-spotted moth to 0.01μg / μL, 0.1μg / μL, 1μg / μL, 10μg / μL and 100μg / μL cis-9-hexadecenal were 15.1±1.1%, 45.2±3.7%, 453.2±56.4%, 502.2±31.7% and 272.4±11.1%, respectively. Figure 3-5 A); the relative values ​​of antennal electrophysiological responses to 0.01μg / μL, 0.1μg / μL, 1μg / μL, 10μg / μL and 100μg / μL concentrations of (Z,Z,Z)-9,12,15-octadecatrienal were 34.3±4.0%, 149.3±12.5%, 348.7±13.3%, 1354.1±58.6% and 466.8±8.3%, respectively.

[0039] The relative values ​​of antennal electrophysiological responses of males of the scarlet hairy moth to cis-9-hexadecenal at concentrations of 1 μg / μL and 10 μg / μL were the highest, significantly higher than those to the treatments of 0.01 μg / μL, 0.1 μg / μL and 100 μg / μL (H4=22.235, P<0.01); the relative values ​​of antennal electrophysiological responses to (Z,Z,Z)-9,12,15-octadecatrienal at a concentration of 10 μg / μL were the highest, significantly higher than those to the treatments of 0.01 μg / μL, 0.1 μg / μL, 1 μg / μL and 100 μg / μL (H4=23.077, P<0.01).

[0040] 1.2 Directional behavioral responses of the red-spotted moth to a single sex pheromone component

[0041] Paraffin was used as a solvent to dilute the two sex pheromone active substances into 5 concentration gradients of 0.01 μg / μL, 0.1 μg / μL, 1 μg / μL, 10 μg / μL and 100 μg / μL, and paraffin was used as a control.

[0042] During the test, 5 μL of the sample to be tested and liquid paraffin were respectively dropped onto 1 cm × 2 cm qualitative filter papers, and were placed in the odor source chambers of the two arms of the "Y" olfactometer, with liquid paraffin as the control. During the test, 5 μL of the sample and liquid paraffin were respectively dropped onto 1 cm × 2 cm qualitative filter papers, and were placed in the odor source chambers (tissue culture bottles) of the two arms of the "Y" olfactometer, with liquid paraffin as the control. Ventilate for 30 seconds to allow the odor to fully flow through the olfactometer. The gas flow rate was controlled at 300 mL min -1 . Only one unmated male of the scarlet hairy moth was released each time. Each male was observed for 5 minutes. Those that entered more than half of the side arm and stayed there for 30 seconds were counted as choosing, otherwise it was recorded as no response and the male was replaced and retested. Each male was used only once. Ten heads were tested in each group, and 5 groups were repeated. After every 5 heads tested, the position of the tube walls on both sides of the olfactometer was swapped to eliminate environmental influences. After each group tested, the olfactometer and connecting tube were cleaned with anhydrous ethanol. The number of males that showed behavioral responses to the treatment and control was recorded.

[0043] Statistical analysis: The single behavioral responses of the scarlet hairy moth to sex pheromones were analyzed using independent sample T-test.

[0044] The results are as follows Figure 2As shown in the figure, the selection rates of males of the red-spotted moth for cis-9-hexadecenal at concentrations of 1 μg / μL, 10 μg / μL and 100 μg / μL were 60.0±3.2% (t8=4.472, P<0.01), 60.0±3.2% (t8=4.472, P<0.01) and 62.0±3.7% (t8=4.536, P<0.01), respectively, while the selection rates for the control were 40.0±3.2%, 40.0±3.2% (t8=4.472, P<0.01) and 62.0±3.7% (t8=4.536, P<0.01), respectively. .0±3.2% and 38.0±3.7%, which were significantly higher than the control liquid paraffin; the selectivity for concentrations of 0.01μg / μL and 0.1μg / μL were 54.0±2.5% (U=4.500, P=0.095) and 50.0±3.2% (t8=1.000, P=1.000), respectively, and the selectivity for the control were 46.0±2.5% and 50.0±3.2%, respectively, with no significant difference between the two groups.

[0045] The selection rates of males of the red-bellied moth for (Z,Z,Z)-9,12,15-octadecatrienal at concentrations of 10 μg / μL and 100 μg / μL were 60.0±3.2% (t8=4.472, P<0.01) and 60.0±5.1% (t8=3.883, P<0.01), respectively, compared with 40.0±3.2% and 40.0±5.1% for the control, respectively, both significantly higher than those for the control. However, the selection rates of males of the red-bellied moth for (Z,Z,Z)-9,12,15-octadecatrienal at concentrations of 0.1 μg / μL and 1 μg / μL were 52.0±3.7% (t8=1.414, P=0.195) and 48.0±3.7% (t8=0.000, P=1.000), respectively, compared with 52.0±3.7% and 48.0±3.7% for the control, respectively, with no significant difference from the control. The selectivity for the drug with a concentration of 0.01 μg / μL was 40.0±4.5%, and the selectivity for the control was 60.0±4.5%, which were significantly lower than those of the control (t8=-3.162, P<0.05).

[0046] Example 2

[0047] Antennae electrophysiological and directional behavioral responses of male Trichoderma vermilion to a mixture of sex pheromones

[0048] A sex pheromone mixture of a reddish-brown moth is prepared, comprising:

[0049] Ratio A: 1 μg / μL cis-9-hexadecenal and 10 μg / μL (Z,Z,Z)-9,12,15-octadecatrienal.

[0050] Ratio B: 1 μg / μL cis-9-hexadecenal and 100 μg / μL (Z,Z,Z)-9,12,15-octadecatrienal.

[0051] Ratio C: 10 μg / μL of cis-9-hexadecenal and 10 μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal.

[0052] Ratio D: 10 μg / μL of cis-9-hexadecenal and 100 μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal.

[0053] Ratio E: 100 μg / μL of cis-9-hexadecenal and 10 μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal.

[0054] Ratio F: 100 μg / μL of cis-9-hexadecenal and 100 μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal.

[0055] 2.1 Antennae electrophysiological responses of male T. vermilionii moths to sex pheromone mixtures

[0056] The optimal concentration of the sex pheromone that elicited a larger relative antennal electrophysiological response and directional behavior in males of the red hairy moth was selected. These two optimal attractant concentrations were mixed to create six mixtures (compositions A to F). The antennal electrophysiological responses of males of the red hairy moth to these mixtures were then measured using the test and analytical methods described in Example 1.

[0057] The results are as follows Figure 3 As shown, the relative values ​​of antennal electrophysiological responses of male C. vermilionii to pheromone combinations A to F were 1449±116.4%, 336.6±12.6%, 71.9±6.3%, 1170±80.7%, 159.3±24.8%, and 418.4±48.5%, respectively. The relative values ​​of antennal electrophysiological responses to combinations A and D were the greatest (F5,24=131.737, P<0.01). These two pheromone combinations were used in the next behavioral experiment.

[0058] 2.2 Orientational behavioral responses of the red-bellied moth to a mixture of sex pheromones

[0059] The directional behavioral responses of male Trichoderma vermilion to the optimal concentration mixtures of two sex pheromones, A and D, were studied using a "Y" type olfactometer.

[0060] During the test, 5 μL of the mixture and paraffin were each dropped onto 1 cm x 2 cm qualitative filter papers. The papers were then placed in the odor source chambers of the two arms of a "Y"-shaped olfactometer, with paraffin serving as a control. The directional behavioral responses of the red-bellied moth to the pheromone mixture were measured according to the experimental and analytical methods described in Example 1.

[0061] The results are as follows Figure 4As shown, the selection rates of males of the scarlet hairy moth for the optimal sex pheromone mixture ratios A and D were 68.0±8.6% (t8=2.959, P<0.05) and 58.0±3.7% (t8=3.024, P<0.05), respectively, while the selection rates for the control were 32.0±8.6% and 42.0±3.7%, respectively, both significantly higher than the control.

[0062] Example 3

[0063] Electrophysiological and directional behavioral responses of male Trichoderma vermilion to single active components of volatiles from Ficus microcarpa

[0064] The single active ingredients of volatiles of Ficus microcarpa include 0.01-100 μg / μL of (-)-α-pinene, 0.01-100 μg / μL of β-myrcene, 0.01-100 μg / μL of β-ocimene and 0.01-100 μg / μL of β-caryophyllene.

[0065] The method for using a single active ingredient of volatiles of Ficus microcarpa is to use liquid paraffin as a solvent to dissolve (-)-α-pinene, β-myrcene, β-ocimene and β-caryophyllene into a solution with a concentration of 0.01 to 100 μg / μL.

[0066] 3.1 Antennae electrophysiological responses of male Trichoderma vermilionii to single active components of volatiles from Ficus microcarpa

[0067] Four active ingredients from volatiles of Ficus microcarpa (Ficus microcarpa)—(-)-α-pinene, β-myrcene, β-ocimene, and β-caryophyllene—were selected. Standard samples of these four active ingredients were prepared at five concentrations, identical to those used for sex pheromones: 0.01 μg / μL, 0.1 μg / μL, 1 μg / μL, 10 μg / μL, and 100 μg / μL. Liquid paraffin was used as the solvent to determine the concentration of each volatile sample that resulted in the maximum antennal electrophysiological response of male trichotomothias vermilionis. The antennal electrophysiological response data of male trichotomothias vermilionis were measured using the experimental and analytical methods described in Example 1.

[0068] like Figure 5 As shown in Figure 3-A, the relative values ​​of antennal electrophysiological responses of male insects to 0.01μg / μL, 0.1μg / μL, 1μg / μL, 10μg / μL and 100μg / μL (-)-α-pinene were 225±13.9%, 36.4±4.9%, 34.9±3.1%, 62.6±4.5% and 618.7±23.4%, respectively. The relative value of antennal electrophysiological response to (-)-α-pinene at 100μg / μL was the highest and significantly higher than the antennal electrophysiological response values ​​at the other four concentrations (F 4，20 =445.564, P<0.01).

[0069] like Figure 5As shown in Figure 2B, the relative values ​​of antennal electrophysiological responses of male insects to 0.01μg / μL, 0.1μg / μL, 1μg / μL, 10μg / μL and 100μg / μL β-myrcene were 22.0±4.6%, 20.0±4.0%, 36.7±6.9%, 628.8±48.5% and 555.4±37.2%, respectively. Among them, the relative values ​​of antennal electrophysiological responses of β-myrcene at 10μg / μL and 100μg / μL were the highest, which were significantly higher than those at the other three concentrations (F 4，20 =234.093, P<0.01).

[0070] like Figure 5 As shown in Figure 3-C, the relative values ​​of antennal electrophysiological responses of male insects to 0.01μg / μL, 0.1μg / μL, 1μg / μL, 10μg / μL and 100μg / μL β-ocimene were 32.8±8.2%, 26.0±7.4%, 35.7±7.3%, 582.5±63.2% and 555.4±37.2%, respectively. The relative values ​​of antennal electrophysiological responses to β-ocimene at concentrations of 10μg / μL and 100μg / μL were the highest, which were significantly higher than those at the other three concentrations (F 4，20 =130.549, P<0.01).

[0071] like Figure 5 As shown in Figure 3-D, the relative values ​​of antennal electrophysiological responses of male insects to 0.01μg / μL, 0.1μg / μL, 1μg / μL, 10μg / μL and 100μg / μL β-caryophyllene were 31.5±4.0%, 57.5±6.5%, 66.9±6.4%, 24.3±4.3% and 216.9±19.8%, respectively. The relative value of antennal electrophysiological response of β-caryophyllene at 100μg / μL was the highest and significantly higher than that at the other four concentrations (F 4，20 =66.332, P<0.01).

[0072] It can be seen that 100 μg / μL of (-)-α-pinene, 10 μg / μL and 100 μg / μL of β-myrcene, 10 μg / μL and 100 μg / μL of β-ocimene, and 100 μg / μL of β-caryophyllene can induce strong antennal electrophysiological responses in male C. vermilionii moth, so these corresponding concentrations of the four active components of volatiles of Ficus microcarpa were used in the next behavioral test.

[0073] 3.2 Directed behavioral responses of males of the red-bellied moth to single active components of volatiles from Ficus microcarpa

[0074] The directional behavioral responses of males of the red-bellied moth were studied using a "Y" type olfactometer.

[0075] During the test, 5 μL of the mixture and paraffin were dropped onto 1 cm x 2 cm qualitative filter papers, respectively. The papers were placed in the odor source chambers of the two arms of a "Y"-shaped olfactometer, with paraffin serving as a control. The behavioral responses of the red-bellied moth to the single active ingredient of Ficus microcarpa were measured according to the experimental and analytical methods described in Example 1.

[0076] like Figure 6 -A, the selection rate of male insects for 100 μg / μL (-)-α-pinene was 62.0±3.7% (t8=4.536, P<0.01), while the selection rate for the control was 38.0±3.7%, which was significantly higher than that of the control; the selection rates for 0.01 μg / μL, 0.1 μg / μL, 1 μg / μL and 10 μg / μL were 52.0±3.7% (t8=0.152, P=0.60 8), 54.0±5.1% (t8=1.109, P=0.299), 52.0±5.8% (t8=0.485, P=0.641) and 52.0±3.7% (t8=0.756, P=0.471), while the selection rates for the control were 48.0±3.7%, 46.0±5.1%, 48.0±5.8% and 48.0±3.7%, respectively, with no significant difference compared with the control.

[0077] like Figure 6 As shown in Figure 2B, the selection rates of male insects for 10 μg / μL and 100 μg / μL β-myrcene were 64.0±4.5% (U=25.000, P<0.01) and 64.8±4.0% (t8=5.228, P<0.01), respectively, while the selection rates for the control were 36.0±4.5% and 35.2±4.0%, respectively, which were significantly higher than the control; the selection rates for 0.01 μg / μL and 0.1 μg / μL were significantly higher than the control. The selection rates of L and 1 μg / μL were 46.4±2.5% (U=20.500, P=0.095), 49.2±4.6% (t8=1.617, P=0.144) and 65±4.0% (t8=5.228, P<0.01), respectively. The selection rates for the control were 53.6±2.5%, 43.9±5.3% and 50.8±4.6%, respectively, with no significant differences compared with the control.

[0078] like Figure 6-C, the selection rates of male insects for 10μg / μL and 100μg / μL β-ocimene were 73.0±4.9% (t8=6.628, P<0.01) and 60.0±3.3% (t8=4.289, P<0.01), respectively, while the selection rates for the control were 27.0±5.3% and 40.0±3.3%, respectively, which were significantly higher than the control; the selection rates for 0.01μg / μL, 0.1μg / μL and The selection rates of 1μg / μL were 57.1±7.7% (t8=1.298, P=0.230), 50.3±7.1% (t8=0.055, P=0.957) and 58.9±5.9% (t8=2.135, P=0.065), respectively. The selection rates for the control were 42.9±7.7%, 49.7±7.1% and 41.1±5.9%, respectively, with no significant difference compared with the control.

[0079] like Figure 6 -D, the selection rates of male insects for 1 μg / μL and 100 μg / μL β-caryophyllene were 70.3±3.2% (t8=9.090, P<0.01) and 58.7±3.8% (t8=3.270, P<0.05), respectively, while the selection rates for the control were 29.7±3.2% and 41.3±3.8%, respectively, which were significantly higher than the control; the selection rates for 0.01 μg / μL, 0.1 μg / μL and 10 The selection rates of μg / μL were 48.0±3.2% (t8=-1.868, P=0.099), 49.6±4.4% (t8=-0.125, P=0.904) and 46.9±2.7% (t8=-1.639, P=0.140), respectively. The selection rates for the control were 52.0±3.2%, 50.4±4.4% and 53.1±2.7%, respectively, with no significant difference compared with the control.

[0080] Based on the results of antennal electrophysiological responses and directional behavioral responses, the antennal electrophysiological responses and directional behavioral responses of the next step were tested using mixed ratios of plant volatiles: 100 μg / μL (-)-α-pinene, 10 μg / μL and 100 μg / μL β-myrcene, 10 μg / μL and 100 μg / μL β-ocimene, and 1 μg / μL and 100 μg / μL β-caryophyllene.

[0081] Example 4

[0082] Antennae electrophysiological and directional behavioral responses of male Trichoderma vermilion to a mixture of active components from volatiles of Ficus microcarpa

[0083] A mixture of active ingredients of volatiles of Ficus microcarpa is prepared, comprising:

[0084] Ratio G: 100 μg / μL of (-)-α-pinene, 10 μg / μL of β-myrcene, 10 μg / μL of β-myrcene β-ocimene and 1 μg / μL of β-caryophyllene.

[0085] Ratio H: 100 μg / μL of (-)-α-pinene, 10 μg / μL of β-myrcene, 10 μg / μL of β-ocimene and 100 μg / μL of β-caryophyllene.

[0086] Ratio I: 100 μg / μL of (-)-α-pinene, 10 μg / μL of β-myrcene, 100 μg / μL of β-ocimene and 1 μg / μL of β-caryophyllene.

[0087] Ratio J: 100 μg / μL of (-)-α-pinene, 10 μg / μL of β-myrcene, 100 μg / μL of β-ocimene and 100 μg / μL of β-caryophyllene.

[0088] Ratio K: 100 μg / μL (-)-α-pinene, 100 μg / μL β-myrcene, 10 μg / μL β-ocimene and 1 μg / μL β-caryophyllene.

[0089] Ratio L: 100 μg / μL of (-)-α-pinene, 100 μg / μL of β-myrcene, 10 μg / μL of β-ocimene and 100 μg / μL of β-caryophyllene.

[0090] Ratio M: 100 μg / μL (-)-α-pinene, 100 μg / μL β-myrcene, 100 μg / μL β-ocimene and 1 μg / μL β-caryophyllene.

[0091] Ratio N: 100 μg / μL of (-)-α-pinene, 100 μg / μL of β-myrcene, 100 μg / μL of β-myrcene β-ocimene and 100 μg / μL of β-caryophyllene.

[0092] 4.1 Antennae electrophysiological responses of male Trichoderma vermilionii to a mixture of active components from volatiles of Ficus microcarpa

[0093] The optimal reaction concentrations of plant volatiles that can induce larger relative antennal electrophysiological response values ​​and directional behavior of male vermilion hairy moths are selected. These four monomer optimal attractant concentrations are mixed to form a total of 8 mixtures (ratios G to N). The antennal electrophysiological responses of male vermilion hairy moths to these mixtures are measured according to the test and analysis methods in Example 1.

[0094] like Figure 7As shown in the figure, the relative values ​​of antennal electrophysiological responses of male insects to eight different concentrations of the four active ingredients of volatiles from Ficus microcarpa were 457.0±34.2%, 279.9±19.7%, 421.4±42.8%, 389.4±44.3%, 306.1±15.9%, 298.2±10.7%, 181.7±8.6% and 217.3±16.7%, respectively. Among them, the relative values ​​of antennal electrophysiological responses of the mixtures G, I and J were the largest (F 7，32 =15.286, P<0.01), so the three active ingredients of volatiles from Ficus microcarpa were mixed and formulated for the next step of the directional behavioral response test.

[0095] 4.2 Oriented behavioral responses of males of the red-bellied moth to a mixture of active components from volatiles of Ficus microcarpa

[0096] A mixture that elicited a greater relative electrophysiological response from the antennae of male Ficus microcarpa was selected and used to study the directional behavioral responses of male Ficus microcarpa moths using a Y-shaped olfactometer. The mixture served as the treatment, while paraffin wax was the control. During the test, 5 μL of the mixture and paraffin wax were each dropped onto a 1 cm x 2 cm qualitative filter paper, each placed within the odor source chamber of the Y-shaped olfactometer. Behavioral responses to the mixture of active ingredients from volatiles of Ficus microcarpa were measured using the experimental and analytical methods described in Example 1.

[0097] like Figure 8 As shown, the selection rates of male C. vermilionii for the optimal mixture ratios of active ingredients of volatiles of Ficus microcarpa G, I and J were 62.8±4.7% (t8=3.886, P<0.01), 62.8±2.0% (t8=8.894, P<0.01) and 60.9±4.4% (t8=3.466, P<0.01), respectively, while the selection rates for the control were 37.2±4.7%, 37.2±2.0% and 39.1±4.4%, respectively, which were significantly higher than the control liquid paraffin.

[0098] Example 5

[0099] Antennae electrophysiological and directional behavioral responses of male Trichoderma vermilion to a mixture of sex pheromones and volatile components from Ficus microcarpa

[0100] The mixture of sex pheromones and active components of volatiles of Ficus microcarpa that showed stronger behavioral responses of males of the red hair moth was selected to form a mixture of plant volatiles and sex pheromones, and a total of 6 mixtures were formed, including:

[0101] Ratio A+G: 1 μg / μL cis-9-hexadecenal, 10 μg / μL (Z,Z,Z)-9,12,15-octadecatrienal, 100 μg / μL (-)-α-pinene, 10 μg / μL β-myrcene, 10 μg / μL β-ocimene and 1 μg / μL β-caryophyllene.

[0102] Ratio A+I: 1 μg / μL cis-9-hexadecenal, 10 μg / μL (Z,Z,Z)-9,12,15-octadecatrienal, 100 μg / μL (-)-α-pinene, 10 μg / μL β-myrcene, 100 μg / μL β-ocimene and 1 μg / μL β-caryophyllene.

[0103] Ratio A+J: 1 μg / μL cis-9-hexadecenal, 10 μg / μL (Z,Z,Z)-9,12,15-octadecatrienal, 100 μg / μL (-)-α-pinene, 10 μg / μL β-myrcene, 100 μg / μL β-ocimene and 100 μg / μL β-caryophyllene.

[0104] Ratio D+G: 10 μg / μL cis-9-hexadecenal, 100 μg / μL (Z,Z,Z)-9,12,15-octadecatrienal, 100 μg / μL (-)-α-pinene, 10 μg / μL β-myrcene, 10 μg / μL β-ocimene and 1 μg / μL β-caryophyllene.

[0105] Ratio D+I: 10 μg / μL cis-9-hexadecenal, 100 μg / μL (Z,Z,Z)-9,12,15-octadecatrienal, 100 μg / μL (-)-α-pinene, 10 μg / μL β-myrcene, 10 μg / μL β-ocimene and 1 μg / μL β-caryophyllene.

[0106] Ratio D+J: 10 μg / μL cis-9-hexadecenal, 100 μg / μL (Z,Z,Z)-9,12,15-octadecatrienal, 100 μg / μL (-)-α-pinene, 10 μg / μL β-myrcene, 100 μg / μL β-ocimene and 100 μg / μL β-caryophyllene.

[0107] 5.1 Behavioral responses to the mixture of sex pheromones and active components of volatiles of Ficus microcarpa

[0108] The antennal electrophysiological response of male Trichoderma vermilion to the above mixture was determined according to the test and analysis methods in Example 1.

[0109] like Figure 9As shown, the relative values ​​of antennal electrophysiological responses of male insects to mixtures A+G, A+I, A+J, D+G, D+I, and D+J were 509.1±53.2%, 280.2±22.0%, 477.8±34.0%, 508.4±18.4%, 308.3±32.4%, and 266.0±29.9%, respectively. Among them, the relative values ​​of antennal electrophysiological responses to mixtures A+G, A+J, and D+G were the largest, significantly higher than those to the other three mixtures (F 5，24 =12.447, P<0.01). Therefore, these three mixtures were used for the next behavioral test.

[0110] 5.2 Behavioral responses to sex pheromones and mixtures of active components from volatiles of Ficus microcarpa

[0111] A Y-shaped olfactometer was used to study the directional behavioral responses of male cristatus moths. A mixture of sex pheromones and plant volatiles was used as the treatment, while paraffin wax served as the control. 5 μL of the mixture and paraffin wax were placed on 1 cm x 2 cm qualitative filter papers, respectively, and placed in the odor source chambers of the Y-shaped olfactometer. Behavioral responses to the mixtures of sex pheromones and plant volatiles were measured according to the experimental and analytical methods described in Example 1.

[0112] like Figure 10 As shown, male insects had a 60.0±3.3% selection rate for the A+G combination (t8 = 4.289, P < 0.01) and a 40.0±3.3% selection rate for the liquid paraffin control. This was significantly higher for the treated group than for the control group (t8 = 4.289, P < 0.01). Male insects had a 60.3±5.0% selection rate for the A+J combination (t8 = 2.956, P < 0.05) and a 39.7±5.0% selection rate for the control group. This was significantly higher for the treated group than for the control group (t8 = 2.956, P < 0.05). Male insects had a 57.0±3.8% selection rate for the D+G combination (t8 = 2.599, P < 0.05) and a 43.0±3.8% selection rate for the control group. This was significantly higher for the treated group than for the control group (t8 = 2.599, P < 0.05). The A+G combination was then used in the next semi-natural baiting experiment.

[0113] Example 6

[0114] Attraction effect of a mixture of sex pheromones and volatile active components from Ficus microcarpa on male Trichoderma vermilion under semi-natural conditions

[0115] A mixture of plant volatiles and sex pheromones (A+G) was used to study the attraction of male larvae of the red-haired moth (Euphrasia vermilion) to a mixture of plant volatiles and sex pheromones under semi-natural conditions using a homemade mesh cage (L×W×H = 1m×1m×1m). The experiment was conducted in an outdoor area with no Ficus plants within a 50-meter radius on clear, windless days, during the peak period of the insect's mate-seeking behavior, between 12:00 PM and 3:00 PM.

[0116] 5 μL of a mixture of plant volatiles and sex pheromones and a single component of the mixture (i.e., a mixture of plant volatiles or a mixture of sex pheromones and paraffin) were respectively dropped onto 1 cm × 2 cm qualitative filter paper. The mixture of plant volatiles and sex pheromones and a single component were placed on the two corners of the upper end of one side of the mesh cage, and two male vermilion hairy moths were placed on the two corners of the lower end of the opposite side. Observe for 10 minutes. If the insects reach the sample and control placement platforms or stay within 5 to 10 cm of them for 30 seconds, they are recorded as having made a choice. Otherwise, they are recorded as not making a choice. Ten insects were tested in each group, and 5 groups were repeated. After each test, the insects were placed on different placement platforms to eliminate environmental interference.

[0117] Statistical analysis: The selection rates of male insects for the three treatment groups under semi-natural conditions were analyzed by one-way ANOVA, and Tukey's HSD was used for multiple comparisons (P<0.05).

[0118] like Figure 11 As shown, under semi-natural conditions, when the control group was liquid paraffin, the selection rate of males of the red-bellied moth for the sex pheromone and volatile active ingredient mixture A+G of Ficus microcarpa was 65.7±5.6%, while the selection rate for the control liquid paraffin was 34.3±5.6%, which were significantly higher than those in the control group (t4=3.952, P<0.05). When the control group was sex pheromone mixture ratio A, the selection rate of males for mixture ratio A+G was 59.3±3.7%, while the selection rate for the control ratio A was 40.7±3.7%, which were significantly higher than those in the control group (t4=3.536, P<0.05). When the control group was volatile active ingredient mixture ratio G of Ficus microcarpa, the selection rate of males for mixture ratio A+G was 59.8±3.5%, while the selection rate for the control ratio G was 40.2±3.5%, which were significantly higher than those in the control group (t4=3.990, P<0.05).

[0119] The above disclosure is only a specific embodiment of the present invention, but the present invention is not limited thereto. Any changes that can be conceived by those skilled in the art should fall within the scope of protection of the present invention.

Claims

1. A red hair moth attractant composition, characterized in that: Its active ingredients are composed of 1μg / μL cis-9-hexadecenal, 10μg / μL (Z,Z,Z)-9,12,15-octadecatrienal, 100μg / μL (-)-α-pinene, 10μg / μL β-myrcene, 10μg / μL β-ocimene and 1μg / μL β-caryophyllene.

2. A red hair moth attractant composition, characterized in that: Its active ingredients are composed of 1μg / μL of cis-9-hexadecenal, 10μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal, 100μg / μL of (-)-α-pinene, 10μg / μL of β-myrcene, 100μg / μL of β-ocimene and 100μg / μL of β-caryophyllene.

3. A red hair moth attractant composition, characterized in that: Its active ingredients are composed of 10 μg / μL of cis-9-hexadecenal, 100 μg / μL of (Z,Z,Z)-9,12,15-octadecatrienal, 100 μg / μL of (-)-α-pinene, 10 μg / μL of β-myrcene, 10 μg / μL of β-ocimene and 1 μg / μL of β-caryophyllene.

4. Use of the attractant composition of any one of claims 1 to 3 in preparing an attractant for the red hairy moth.

5. Use of the attractant composition of any one of claims 1 to 3 for the control and population monitoring of the red hairy moth.

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