CN117356566A - Application of (R) -Lavender (R) -2-methylbutyrate in pheromone attractant and attracting core - Google Patents

Abstract

The invention belongs to the technical field of plant protection agents, and relates to application of (R) -Lavender (R) -2-methyl butyrate in an pheromone attractant and an attracting core. According to the invention, the solid-phase microextraction technology is adopted to extract the body surface volatile of the frankliniella occidentalis, the GC-MS analysis is adopted to identify that the body surface volatile of the frankliniella occidentalis consists of (R) -lavender acyl (R) -2-methyl butyrate, the attraction of the (R) -lavender acyl (R) -2-methyl butyrate to female and male adult frankliniella occidentalis is further verified through an indoor Y-type olfactometer test, the pheromone attractant component of the frankliniella occidentalis is defined, and the method can be used for dynamic detection of pest population and prediction and forecast of occurrence rules, and provides technical support for effective prevention and control of the frankliniella occidentalis.

Description

Application and application of (R)-lavenderoyl (R)-2-methylbutyrate in pheromone attractants Lure

Technical field

The invention belongs to the technical field of plant protection agents and relates to the application of (R)-lavenderoyl (R)-2-methylbutyrate in pheromone attractants and lure cores.

Background technique

Thrips are the collective name for insects of the order Thysanoptera. They have many species, a wide distribution range, and serious harm. They are one of the most important agricultural pests in the world. Their strong fecundity, concealment, and drug resistance make their prevention and control difficult. It is a major bottleneck restricting the high-quality development of the agricultural industry. Odontothrips loti Haliday is a dominant pest that harms alfalfa. It is widely distributed in major alfalfa planting areas such as Xinjiang, Inner Mongolia, and Ningxia. It feeds on the above-ground parts of plants in its larvae and adult stages and can spread plant virus diseases, causing plant damage. The leaves are yellow and withered.

At present, there are three main methods for thrips control: chemical control, biological control and physical control. However, because thrips are small, easy to hide, and highly active, most sticky insect boards and insect traps have little effect on attracting thrips. It is not obvious, attracts non-target insects, and the cost is high. It is easy to develop resistance to most chemical pesticides, the pesticide control effect is reduced, causing serious pollution to the environment, posing a serious threat to food safety, affecting human health, leading to chemical The effects of prevention and physical control are not ideal. In view of this current situation, it is an inevitable trend to find a pollution-free and sustainable biological control method of thrips to solve the harm to health and environment caused by existing technologies, and using the pheromones of insects themselves to control thrips is one of the effective measures.

Currently, it is known that male adults of various thrips release aggregation pheromone, which can function as a single component or as a mixture of multiple components released in a certain proportion. It can significantly attract adults of both sexes and is an important substance for information exchange among thrips populations. . In the existing technology, the following aggregation pheromone components that attract thrips have been effectively separated and identified. CN108902143B discloses a thrips aggregation pheromone which is geranyl acetate and farnesyl acetate. CN104381256B discloses a reagent for attracting flower thrips, which is (R)-lavender acetate, and stilbene-based (S)-2-methylbutyrate or stilbene-based (S)-2-methylbutyric acid. The mass ratio of the ester analogs isobornyl valerate, isobornyl 2-methylbutyrate, isobornyl trimethylacetate, and lavender isovalerate is 1:8, which has the strongest attracting ability. CN109730065B discloses an attractant composition for western flower thrips, which is (R)-lavender acetate and acenaphthyl (S)-2-methylbutyrate. CN108041034B discloses an aggregation pheromone attractant for female tea stick thrips and a method for preparing the lure core of the attractant, wherein the aggregation pheromone attractant is 2,4,7,9-tetramethyl-5-decyne- 4,7-diol.

The control of flower thrips aggregation pheromone has become a model for the application of insect aggregation pheromone. Therefore, studying the aggregation pheromone of T. truncatula and developing related application technologies can be widely used in pest prediction, trapping, interfering with mating, etc., in order to monitor and concentrate the population monitoring of T. truncatula. Trapping prevention and control provides an effective method, and at the same time, it can effectively protect alfalfa plant varieties, alleviate problems such as increased resistance and incomplete prevention caused by chemical pesticide control, thereby avoiding serious pollution to the environment and serious threats to food safety, thus affecting human health.

Contents of the invention

The object of the present invention is to provide the application and lure core of (R)-lavenderoyl (R)-2-methylbutyrate in pheromone attractants. The present invention uses solid-phase microextraction technology to extract volatiles from the body surface of Trichoderma thrips, and uses Shimadzu GC-MS QP2010 PLUS (gas chromatography-mass spectrometry, GC-MS) to separate and identify the components of the volatiles. The attractant effect was further verified through an indoor Y-type olfactometer test, the components of the pheromone attractant of T. truncatula were clarified, and an aggregation pheromone with significant attracting effect on male and female T. truncatula was developed. It provides technical support for the effective prevention and control of horn-toothed thrips.

Based on the above purpose, the present invention solves this need in the field by providing the application of (R)-lavenderoyl (R)-2-methylbutyrate in pheromone attractants and lure cores.

In one aspect, the invention relates to the use of (R)-lavandμLyl(R)-2-methylbutanoate as a pheromone attractant.

Preferably, the application of (R)-lavenderoyl (R)-2-methylbutyrate provided by the present invention in pheromone attractants includes the pheromone attractant (R)-lavenderoyl (R)-2 - Methylbutyrate is used to attract male and female adults of A. striata.

The molecular structural formula of the (R)-lavenderoyl (R)-2-methylbutyrate is as follows;

Preferably, the application of (R)-lavenderoyl (R)-2-methylbutyrate provided by the present invention in pheromone attractants includes the pheromone attractant (R)-lavenderoyl (R)-2 - Methylbutyrate attractant concentration is 0.5μg/μL-2μg/μL.

On the other hand, the present invention relates to a lure for attracting Aphrodisiac thrips, the lure comprising: (R)-lavenderoyl (R)-2-methylbutyrate and paraffin oil.

On the other hand, the present invention relates to a lure for attracting Aphrodisiac thrips, in which (R)-lavenderoyl (R)-2-methylbutyrate is diluted to 0.5 with paraffin oil. μg/μL-2μg/μL.

Compared with the prior art, the present invention has the following beneficial effects or advantages:

The thrips is the main pest that harms alfalfa. The present invention provides a pheromone attractant that can be used for the thrips, which can effectively attract both sexes of the thrips. It is prepared by using it simultaneously with paraffin oil. The lure core can effectively attract the thrips in alfalfa. It has excellent attracting effect, that is, it has high sensitivity. It can be used as one of the methods for the quarantine department to improve the identification of thrips. It can be used for Dynamic detection of pest populations, prediction of occurrence patterns, and concentrated trapping of pests provide technical support for the effective prevention and control of thrips, thereby achieving the goal of ensuring and increasing production.

Description of the drawings

In order to explain the embodiments of the present invention or the technical solutions in the prior art more clearly, the drawings needed to be used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings in the following description are only are some embodiments of the invention.

Figure 1 shows the total ion chromatogram and mass spectrum of the volatiles on the body surface of the male body of Thrips angustifolia. A: Total ion chromatogram of volatiles from the male body surface of the male body surface of Trichoderma thrips; B: Partial enlargement of the peak time of 27.37 minutes.

Figure 2 shows the comparison results between the male body surface volatile matter spectrum and the NIST mass spectrum library.

Figure 3 shows the behavioral response of adult Thrips thrips to lavenderoyl-2-methylbutyrate isomers.

Figure 4 shows the behavioral response of adult Thrips thrips to lures prepared from (R)-lavenderoyl (R)-2-methylbutyrate and paraffin oil.

Detailed ways

The technical solutions of the present invention will be described below with reference to examples. However, the present invention is not limited to the following examples. The experimental methods and detection methods described in each example are conventional methods unless otherwise specified. The equipment and raw materials mentioned above can be purchased in the market unless otherwise specified.

The invention provides a kind of pheromone attractant for A. thrips. Its (R)-lavenderoyl (R)-2-methylbutyrate (purity is 90%) and other enantiomers are produced by China Agriculture It was synthesized in the laboratory of Bian Qinghua, Department of Chemistry, School of Science, University, and diluted to 0.5 μg/μL, 1 μg/μL and 2 μg/μL with solvent liquid paraffin oil (analytical grade, S10187, SCRC).

Example 1

1. Materials and methods

This embodiment provides a method for the isolation, identification and application of volatiles on the body surface of Thrips angustifolia.

1.1 The origin of thrips

The thrips genus were collected from the Shangzhuang Experimental Station of China Agricultural University. The population was established in a laboratory artificial incubator using fresh alfalfa leaves and lentils for multiple generations. The ambient temperature is 26±1℃, the humidity is 60%, and the light L:D=16:8h.

1.2 Use solid-phase microextraction (SPME) technology to collect volatiles from the body surface of A. thrips

Step 1: The solid-phase microextraction adsorption needle needs to be inserted into the injection port of the gas chromatograph for aging for 30 minutes when used for the first time;

Step 2: Place 10 active male adults into a 5ml sample bottle, then insert the adsorption needle into the sample bottle, rotate and extend the adsorption probe to collect the collected samples for 30 minutes;

Step 3: Repeat 3 times, 10 test insects/time, and all test insects are only used for one collection.

1.3 Separation process:

The collected volatiles were separated and identified by Shimadzu GC-MS QP2010 PLUS. The chromatographic column selected is Rtx-5MS (30.0m×0.25mm×0.25μm);

The total duration of the GC program for the identification of volatiles on the body surface of Thrips angustifolia is 32.5 minutes. The starting temperature is 45°C, held for 1 minute, raised to 150°C at a rate of 5°C/min, held for 1 minute, and then raised at a rate of 10°C/min. to 200°C and hold for 1 minute; then increase the temperature to 250°C at a rate of 20°C/min and hold for 1 minute. The inlet temperature was 250°C, splitless injection was used; the carrier gas was helium, and the flow rate was 1 ml/min.

1.4 Identification of chemical components and structural analysis of aggregation pheromones

According to the total ion chromatogram, determine the peak position of the target compound, then compare the target peak mass spectrum with the NIST online database, and select the most matching result based on the degree of matching between the search matching degree and the mass spectrum peak type and characteristic ion peak information, and conduct a preliminary Qualitative.

1.5 Use a Y-type olfactometer to measure the selection behavior of lavender yl-2-methylbutyrate isomers by male and female adult Thrips thrips.

Dilution of test sample:

(R)-lavenderoyl (R)-2-methylbutyrate and other enantiomeric purity reached 90%, synthesized by Bian Qinghua Laboratory, Department of Chemistry, College of Science, China Agricultural University, using solvent liquid paraffin oil ( Analytical grade, S10187, SCRC), (R)-lavenderoyl (R)-2-methylbutyrate and other enantiomers, diluted to 0.5μg/μL, take 50μL test samples and put them into the sample loading bottle, and add 50 μL of liquid paraffin oil to each isomer of (R)-lavenderoyl(R)-2-methylbutyrate as a control.

Y-type olfactometer measurement method:

The Y-shaped tube is made of transparent glass. The length of the main tube and the two arms are 11cm, the inner diameter is 1cm, and the angle between the two arms is 75°. The Y-shaped olfactometer is connected to the Y-shaped tube, glass rotor flowmeter (LZB-3WB), and sample loading bottle (100ml) in sequence. , distilled water (gas washing bottle 250ml), activated carbon drying tower and air sampler. The interfaces are all connected with Teflon tubes. The air sampler controls the flow rate to 50ml/min. Set up two sample loading bottles at each dose and add 50 μL of (R)-lavenderoyl (R)-2-methylbutyrate sample and its enantiomer and 50 μL of control sample (liquid paraffin oil) as odor source. An LED light panel (28W) is placed at the Y end of the Y-type olfactometer as the only induction light source. The test environment temperature is maintained at 26±1°C. The test is completed between 10:00-17:00 during the day.

For thrips, select 3-5 day old female and male adults, introduce the test thrips head by head into the center of the main tube of the Y-shaped tube, and record the number that enters both arms of the olfactometer. The timer starts when the thrips are introduced into the main tube. If the thrips reach 1/2 of the two arms within 5 minutes and stay for more than 10 seconds or more than 2/3 of the arms, it is deemed to have made a selection of the taste source, otherwise it is considered to have not responded. To avoid position effects, the odor sources were swapped after testing five thrips. To avoid the influence of the residual odor of test thrips, a new Y-tube was replaced after testing 10 thrips. The Y-tube was cleaned with 75% ethanol and baked at 60°C for 30 minutes. The experiment was repeated 60 times, and all test insects were used only once.

1.6 Use a Y-type olfactory instrument to measure the selection behavior of male and female adults of Aphrodisiac thrips on lures prepared from (R)-lavenderoyl (R)-2-methylbutyrate and paraffin oil.

Preparation of a lure for attracting horn-toothed thrips, including the following process steps:

Step 1: Preparation of sustained-release carrier

Choose green natural rubber as the lure carrier, soak it in anhydrous ethanol for 24-48 hours, take out the rubber plug and put it in an oven at 30-50°C for 24-48 hours;

Step 2: Configuration of Aggregated Pheromone Attractants

A lure core composed of (R)-lavenderoyl (R)-2-methylbutyrate and paraffin oil was prepared. A total of 3 test groups were set up. The lure core was prepared by adding (R)-lavenderoyl (R)-2-methylbutyrate with paraffin oil. )-2-methylbutyrate was diluted to 0.5 μg/μL, 1 μg/μL, and 2 μg/μL respectively. For each test group, 50 μL paraffin oil was added as a control.

Step 3: Prepare the lure

The lure carrier is immersed in the bottle containing the aggregated pheromone attractant prepared in step 2, soaked for 24 hours, then taken out, and allowed to air dry naturally in a cool indoor place. The control group was soaked in paraffin oil only. The completed lures are wrapped in tin foil and placed in a -20°C refrigerator for later use.

The Y-type olfactometer measurement method is the same as the Y-type olfactometer measurement method in 1.5.

1.7 Data analysis

Chi-squared test (Chi-squared) was used to analyze behavioral choices. Other statistical analyzes were completed using office2010 and SPSS22.0, and Adobe Photoshop CC2018 was used for drawing.

2. Results and analysis

2.1 Identification of chemical components of aggregation pheromone

The results in Figure 1 show that a substance was found in the total ion chromatogram of male adult thrips, with a peak time of 27.37 minutes. The results in Figure 2 show that the mass spectrum of this component was compared with the NIST mass spectrum library, and the results showed that the substance with the highest similarity was (R)-lavenderoyl (R)-2-methylbutyrate, with a similarity of 95%. Since (R)-lavenderoyl (R)-2-methylbutyrate has two chiral carbon atoms, there are 4 enantiomers, and the volatile substance can be identified as (R)-lavenderoyl ( R)-2-methylbutyrate and its other enantiomers, in order to determine the specific structure of its volatiles, a Y-type olfactory instrument was used to determine the relationship between (R)-lavenderoyl (R) by A. -2-Methylbutyrate and its other enantiomers behavior selection test results were analyzed.

2.2 Results of the behavioral response of Phyllostachys thrips to (R)-lavenderoyl (R)-2-methylbutyrate isomers

Since (R)-lavenderoyl(R)-2-methylbutyrate has two chiral carbon atoms, there are 4 enantiomers. We compare these 4 enantiomers and racemic Behavioral verification was conducted on the body. The results in Figure 3 show that at a concentration of 0.5 μg/μL, only (R)-lavenderoyl (R)-2-methylbutyrate showed a significant attraction to male adults. Qualitative horn The volatile compound on the body surface of thrips is (R)-lavenderoyl (R)-2-methylbutyrate.

2.3 Results of the behavioral response of Phyllostachys thrips to lures prepared from (R)-lavenderoyl (R)-2-methylbutyrate and paraffin oil

Figure 4 shows the results of the behavioral selection test of Phyllostachys thrips on lures prepared with (R)-lavenderoyl (R)-2-methylbutyrate and paraffin oil. The results in Figure 4 show that the lure prepared by (R)-lavenderoyl (R)-2-methylbutyrate and paraffin oil exhibits strong attraction to the male adult pair of T. Acyl(R)-2-methylbutyrate was diluted with paraffin oil at three concentration gradients of 0.5 μg/μL, 1 μg/μL, and 2 μg/μL, and showed strong attraction to male and female adults of A. , and as the concentration increases, the attraction effect is stronger, and male adults respond more strongly than female adults.

When the concentration of (R)-lavenderoyl (R)-2-methylbutyrate diluted with paraffin oil was 0.5 μg/μL, the prepared lure showed significant attraction to male adults, but the difference was not significant for female adults. When the concentration of (R)-lavenderoyl (R)-2-methylbutyrate diluted with paraffin oil was 1 μg/μL, the prepared lure showed significant attraction to male adults, but the difference was not significant for female adults. When the concentration of (R)-lavenderoyl (R)-2-methylbutyrate is diluted with paraffin oil to 2 μg/μL, the prepared lure performs well on unmated female adults, female adults and male adults of A. There was a significant attraction, indicating that the lure prepared by (R)-lavenderoyl (R)-2-methylbutyrate and paraffin oil was used to convert (R)-lavenderoyl (R)-2-methylbutyrate into When the dilution concentration of the acid ester is 2 μg/μL, the attraction effect is the best, which provides technical support for the preparation of pheromone attractants for attracting horny thrips.

As described above, the present invention can be better implemented. The above-mentioned embodiments are only descriptions of preferred embodiments of the present invention and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, ordinary people in the art can Various changes and improvements made by skilled personnel to the technical solution of the present invention should fall within the protection scope of the present invention.

Claims (5)

1. Application of (R)-lavenderoyl (R)-2-methylbutyrate in pheromone attractants. 2. The application according to claim 1, characterized in that it is used for attracting male and female adults of Phytophthora thrips. 3. The application according to claim 2, characterized in that the attractant concentration of (R)-lavenderoyl (R)-2-methylbutyrate is 0.5 μg/μL-2 μg/μL. 4. A lure for attracting Aphrodisiac thrips, characterized in that the lure includes: (R)-lavenderoyl (R)-2-methylbutyrate and paraffin oil. 5. The lure core according to claim 4, characterized in that (R)-lavenderoyl (R)-2-methylbutyrate is diluted to 0.5 μg/μL-2 μg with paraffin oil in the lure core. /μL.