CN117397681B - A wolfberry fruit fly sex pheromone and its application - Google Patents

Abstract

The invention relates to a wolfberry fruit fly sex pheromone and its application. Through GC-MS analysis of volatiles released by Lycium barbarum fruit fly and identification of volatiles secreted by the gonads of Lycium barbarum fruit fly, the present invention determines that the sex pheromone secreted by female adults of Lycium barbarum fruit fly is triallyl carbinol. Using triallyl carbinol as a sex attractant to attract wolfberry fruit flies can achieve the purpose of green prevention and control.

Description

A wolfberry fruit fly sex pheromone and its application

Technical field

The present invention relates to the field of pest control, and in particular to an attractant.

Background technique

The wolfberry fruit fly (Neoceratitis asiatica) is an important and disastrous pest that damages wolfberry fruits. The female adult has a sharp ovipositor that can be inserted into the wolfberry fruit to lay eggs. The wound forms a brown milky protrusion. After the larvae hatch, they can feed on the fresh fruit pulp. As the larvae grow, white curved markings appear on the surface of the fruit, forming "Maggot fruit" loses its economic and medicinal value, which seriously affects the yield and quality of wolfberry.

When the pregnant female wolfberry fruit fly begins to lay eggs in the fruit, it is the primary stage of damage, and the courtship and mating stages are the key periods to actually control the large outbreak of wolfberry fruit fly. Sex pheromone is a key chemical substance that is secreted by male and female insects and then received by the opposite sex and triggers courtship and mating behavioral responses. Sex attractants developed based on it have a good control effect on insects.

Contents of the invention

One aspect of the present invention provides the use of triallyl carbinol as a sex pheromone of Lycium barbarum fruit fly (Neoceratitis asiatica).

In a specific embodiment, the application is for attracting wolfberry fruit flies.

In a specific embodiment, the Lycium barbarum fruit fly is a male adult of Lycium barbarum fruit fly.

The second aspect of the present invention provides the use of a composition containing triallyl carbinol for attracting wolfberry fruit flies.

In a specific embodiment, the composition further includes a pesticide acceptable carrier.

In a specific embodiment, the Lycium barbarum fruit fly is a male adult of Lycium barbarum fruit fly.

The third aspect of the present invention provides a lure, which includes triallyl carbinol and a sustained-release carrier.

In a specific embodiment, the material of the sustained-release carrier is rubber.

In a specific embodiment, the sustained-release carrier is at least one of a rubber-tipped carrier, a cotton core carrier, and a microtube carrier.

The fourth aspect of the present invention provides the application of the lure core according to the third aspect of the present invention for luring wolfberry fruit flies.

In a specific embodiment, the Lycium barbarum fruit fly is a male adult of Lycium barbarum fruit fly.

Beneficial effects of the present invention: The present invention has discovered for the first time that triallyl carbinol is the main component of the sex pheromone secreted by female adults of Lycium barbarum fruit fly. It can be used to attract male adults of Lycium barbarum fruit fly. It can be used as a sex attractant to achieve green color. prevention and control purposes.

Description of drawings

Figure 1 shows the mass spectrometry results of the specific release of triallyl carbinol from female adults of Lycium barbarum fruit fly.

Figure 2 shows the EAG measurement results of Lycium barbarum fruit fly males at various concentrations of triallyl methanol solution.

Figure 3 shows the EAG measurement results of male and female fruit flies of Lycium barbarum at a concentration of 10 ^-3 M in triallyl methanol solution.

Figure 4 shows the Y-type olfactometer measurement results of Lycium barbarum fruit fly males at various concentrations of triallyl methanol solution. Among them, the value outside the brackets is the amount induced by n-hexane, and the value inside the brackets is the amount induced by triallylmethanol.

Figure 5 shows the Y-type olfactometer measurement results of male and female fruit flies of Lycium barbarum at a concentration of 10 ^-3 M in triallyl methanol solution. Among them, the value outside the brackets is the amount induced by n-hexane, and the value inside the brackets is the amount induced by triallylmethanol.

Figure 6 shows the trapping effect of the lure on male adults of Lycium barbarum fruit fly.

Detailed ways

The above content of the present invention will be further described in detail below in the form of preferred embodiments, but this does not constitute a limitation of the present invention.

Unless otherwise specified, the reagents in the examples of the present invention can be purchased through commercial channels.

All statistical comparisons were analyzed using SPSS 22.0 software (SPSS Inc., Chicago, IL, USA).

Multiple comparisons of data were performed using Turkey's HSD test in one-way analysis of variance ( P <0.05).

The independent sample Student’s t test was used to analyze statistical differences between the two treatments.

Statistical differences in Y-type olfactometer experimental results were analyzed using the two-sided binomial test method.

All statistical analysis figures were generated using GraphPad Prism 6.01 software (San Diego, CA).

Insect rearing.

From May to July 2022, wolfberry fruit fly pupae were collected from the Zhongning wolfberry planting base in the Ningxia Hui Autonomous Region (the wolfberry variety is Ningqi No. 1, and the tree age is 10 years old). They were cultured in an artificial climate chamber (Ningbo Saifu Experimental Instrument Co., Ltd., PRX-450 C).

The feeding conditions for wolfberry fruit flies are: photoperiod 14 L: 10 D, temperature (25 ± 2)°C, and relative humidity 40% ±10%. Put the wolfberry fruit fly pupae into a petri dish (3 cm high, 20 cm diameter) filled with about 3 cm of sandy loam soil, and then place it in a mesh cage (50 cm long × 50 cm wide × 50 cm high) for culture. After adults emerged, unmated female and male adults were placed in different mesh cages (50 cm long × 50 cm wide × 50 cm high) and fed a mixture of yeast extract powder and sucrose (1:3).

Collection and GC-MS analysis of volatiles released by Lycium barbarum fruit fly.

Collection of volatiles released by Lycium barbarum fruit fly: The dynamic headspace adsorption method was used to collect volatiles from female and male adults of Lycium barbarum fruit fly. The specific method is as follows: Put the fruit flies on the 1st, 2nd, 3rd, 4th and 5th days after emergence into 2 L headspace collection glass bottles (100 fruit flies in each bottle). Connect it to the adsorption tube containing the adsorbent, and connect the other end to the glass bottle containing activated carbon. Start the air sampler. The fruit fly odor flows into the adsorbent with the air flow. The collection air flow is 300 mL/min. After 6 hours of collection, , use chromatographically pure n-hexane to elute and elute the adsorbent into a sample bottle, and store it at -20°C. Each experiment was set up with 3 biological replicates.

Identification of volatiles released by Lycium barbarum fruit fly: Gas chromatography mass spectrometry GC-MS was used to identify the components of volatiles released by male and female Lycium barbarum fruit flies. Set the parameters on the machine (the flow rate is 1.0 mL/min, the ionization voltage and emission current of the electron impact mass spectrometer are 70 eV and 34.6 μA respectively, the temperatures of the ion source and interface are 230 °C and 280 °C respectively), after starting Automatic injection, 4 μL per injection. The database was used to search for mass spectra and chromatogram peak areas for qualitative and quantitative analysis of compounds. Each experiment was set up with 3 biological replicates and 3 technical replicates. Comparing the volatiles released by female adults of Lycium barbarum fruit flies with those released by male adults, it was found that female adults specifically release triallyl carbinol (alias: 4-allyl-hept-4-ol), whose molecular formula is C ₁₀ H ₁₆ O, while male adults did not release triallyl carbinol. The results are shown in Figure 1.

Furthermore, the volatiles secreted by the gonads of female adults of Lycium barbarum were identified, and it was determined that triallyl carbinol is secreted by the gonads of female adults of Lycium barbarum.

In summary, triallyl carbinol is a sex pheromone secreted by female adults of Lycium barbarum fruit fly.

Electrophysiological and behavioral responses of Lycium barbarum fruit flies to triallyl carbinol.

Electroantennography (EAG) and Y-type olfactometer were used to test the electrophysiological responses and behavioral responses of female and male adults of Lycium barbarum fruit fly to different concentrations of triallyl carbinol standards.

Antenna potential measurement system EAG test: Before the experiment, cut off the head of the wolfberry fruit fly, connect the top of its antennae to one electrode, and the head to the other electrode. The distance between the antennae and the odor tube on the fixed frame should not exceed 1 cm. Take 10 μL of the odor source (10 ^-1 M, 10 ^-2 M, 10 ^-3 M and 10 ^-4 M triallyl methanol solutions prepared with n-hexane solvent) and drop it on a 5 cm × 1 cm filter paper strip. on, and then put the filter paper strip into a 10 cm long air tube, and the end of the air tube is connected to the odor stimulation controller. When a stable electrical signal appears, odor stimulation is given, airflow is sent from the stimulation controller at 50 cm/s, the pulse time is maintained at 2 s, and the EAG signal is observed. The time interval between two odor stimulations was 1 min to ensure the recovery of the antennal potential. Ten male and female wolfberry fruit flies were provided in the experiment. Each antennae was stimulated three times with a single odor source, and n-hexane was used as the control group. The difference between the average value of the observed values on the same antenna and the values measured before and after in the control group was used as standardization. The measurement results of Lycium barbarum fruit fly males at various concentrations are shown in Figure 2. The results of the determination of male and female fruit flies of Lycium barbarum at a concentration of 10 ^-3 M are shown in 3.

According to the results in Figure 2, it can be seen that the male wolfberry fruit fly has a strong electrophysiological response to 10 ^-2 M and 10 ^-3 M triallyl carbinol solutions.

According to the results in Figure 3, it can be seen that the electrophysiological response of male Lycium barbarum fruit flies to triallyl carbinol is significantly higher than that of female flies.

Y-type olfactometer test: Take 10 μL of odor source (10 ^-1 M, 10 ^-2 M, 10 ^-3 M and 10 ^-4 M triallyl methanol solutions prepared with n-hexane solvent) and drop them on a clean Put the end of one arm of the Y-type olfactory meter on the filter paper (1 cm In the instrument arm, the air flow is 300mL/min. Place the full and unmated male or female fruit fly of Lycium barbarum in the main arm of the olfactometer, observe their behavioral choices, and count the number of insects selected at each odor concentration. Each experiment was repeated three times with 60 fruit flies. The measurement results of Lycium barbarum fruit fly males at various concentrations are shown in Figure 4. The results of the determination of male and female fruit flies of Lycium barbarum at a concentration of 10 ^-3 M are shown in 5.

According to the results in Figure 4, it can be seen that 10 ^-2 M and 10 ^-3 M triallyl methanol solutions have a strong attracting effect on male wolfberry fruit flies.

According to the results in Figure 5, it can be seen that the triallylmethanol solution with 10 ^-3 M has no attracting effect on the female wolfberry fruit fly.

Application of triallylcarbinol-based lures to Lycium barbarum fruit fly.

Based on the behavioral response of triallyl carbinol in the concentration range of 1/100 to 1/1000 M, seven lures with different contents of triallyl carbinol sustained-release carriers (rubber-type carriers) were designed, respectively 1/50 M, 1/100 M, 1/200 M, 1/500 M, 1/1000 M and 1/2000 M, prepared with n-hexane to obtain triallylmethanol solutions at each concentration. Take 100 μL of triallyl carbinol solution and add it dropwise to the sustained-release carrier. After natural volatilization, the treatment lure core at each dosage is prepared. The control lure was used as a control lure after adding dropwise 100 μL of n-hexane and evaporating it. Among them, the dosage of triallylcarbinol at 1/50M concentration is 304.46 μg, the dosage of triallylcarbinol at 1/100M concentration is 152.23μg, and the dosage of triallylcarbinol at 1/200M concentration is 76.13 μg, the dosage of triallyl carbinol at 1/500 M concentration is 30.45 μg, the dosage of triallyl carbinol at 1/1000 M concentration is 15.22 μg, and the dosage of triallyl carbinol at 1/2000 M concentration is 76.13 μg. The amount of methanol used was 7.61 μg.

Place the lure on an adhesive yellow board and hang it on the side wall of a cage made of 50 cm × 50 cm gauze. After 48 hours, count the number of male adult fruit fly traps. The results are shown in Figure 6.

According to the results in Figure 6, it can be seen that the lures treated with triallyl carbinol at 15.22 to 152.23 μg attracted significantly higher adult male wolfberry fruit flies than the control lures (n-hexane treated lures).

Claims (7)

1. Application of triallyl carbinol in attracting male adults of Lycium barbarum fruit fly. 2. Application of a composition containing triallylcarbinol for luring Lycium barbarum fruit fly, which is a male adult of Lycium barbarum fruit fly. 3. The application according to claim 2, characterized in that the composition further includes a pesticide acceptable carrier. 4. A lure, which includes triallyl carbinol and a sustained-release carrier. 5. The lure core according to claim 4, characterized in that the material of the sustained-release carrier is rubber. 6. The lure core according to claim 4, characterized in that the sustained-release carrier is at least one of a rubber-tipped carrier, a cotton core carrier and a microtube carrier. 7. Application of the lure according to any one of claims 4 to 6 in luring wolfberry fruit flies, which are wolfberry fruit fly male adults.