A chinese trichoplusia ni attractant and its preparation method and application

Abstract

The application provides a Chinese trichopoda japonica attractant and a preparation method and application thereof, and belongs to the technical field of biological pest control. The active ingredient of the Chinese trichopoda japonica attractant is composed of volatile substances of potentilla chinensis, and the volatile substances include (Z)-3-hexen-1-ol 10-15 parts, gamma-terpinene 10-20 parts, trans-beta-myrcene 60-70 parts and nerolidol 0.5-1 part in volume fraction. The Chinese trichopoda japonica attractant as the effective component is combined with a diluent to further obtain an attractant preparation. The volatile substances in the potentilla chinensis are used to perform olfactory behavior response on the Chinese trichopoda japonica and are applied, the obtained attractant or attractant preparation has obvious attracting effect, is harmless to other natural enemies and is environment-friendly, and provides a new reference means for biological control.

Description

Technical Field

[0001] This invention belongs to the field of biological attractant development technology, and particularly relates to an attractant for the Chinese lacewing, its preparation method and application. Background Technology

[0002] Predatory natural enemies are important consumers in ecosystems, balancing the entire ecosystem and influencing the overall community structure and density of pests. The larvae and adults of the Chinese lacewing (Chrysopa sinica) prey on many pests in various economic crop fields, making it a dominant predator. Furthermore, it is widely distributed, has a strong predatory ability, consumes a large amount of food, reproduces rapidly, and has a long occurrence period, giving it considerable potential for biological control. The Chinese lacewing feeds on many crop pests (vegetables, fruits, cotton), such as aphids (wheat, cotton, corn aphids, etc.), and can also prey on whiteflies and thrips.

[0003] Biological control has become an important means of pest control, and intercropping functional plants can provide natural enemies with alternative food, hosts, prey, and shelter. Cnidium monnieri, an annual plant of the Apiaceae family, attracts natural enemies such as ladybugs, lacewings, predatory bugs, and hoverflies. One reason for the increase in natural enemies due to Cnidium monnieri is its volatile organic compounds (VOCs). Cnidium monnieri can produce various secondary metabolites, including terpenoids, esters, and alcohols. However, in complex environments, only a small portion of these VOCs affect the behavior of natural enemies. Therefore, this invention proposes a lacewing attractant, its preparation method, and its application. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention proposes an attractant for the Chinese lacewing, its preparation method, and its application.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] One of the technical solutions of the present invention:

[0007] A type of attractant for the Chinese lacewing, the active ingredient of which is composed of volatile substances from Cnidium monnieri, including (Z)-3-hexen-1-ol, γ-terpinene, trans-β-ocimene, and nerolidol.

[0008] There are various volatile substances that attract the Chinese lacewing. This invention obtained the above-mentioned Chinese lacewing attractant through multiple screenings and combinations. Further research found that the amount of each volatile substance directly determines the attraction effect of the Chinese lacewing attractant. As a preferred embodiment, the Chinese lacewing attractant of this invention includes, by volume, 10-15 parts of (Z)-3-hexen-1-ol, 10-20 parts of γ-terpinene, 60-70 parts of trans-β-ocimene, and 0.5-1 parts of nerolidol.

[0009] More preferably, by volume parts, it comprises: 12.9 parts of (Z)-3-hexen-1-ol, 18.8 parts of γ-terpinene, 67.6 parts of trans-β-ocimene, and 0.7 parts of nerolidol.

[0010] The second technical solution of the present invention:

[0011] A method for preparing the attractant for *Tetrapanax papyrifer* described above, comprising weighing (Z)-3-hexen-1-ol, γ-terpinene, trans-β-ocimene, and nerolidol by volume, and mixing them evenly.

[0012] The third technical solution of the present invention:

[0013] A *Tetrapanax papyrifer* attractant formulation comprises an active ingredient and a diluent, wherein the active ingredient is the *Tetrapanax papyrifer* attractant, and the volume ratio of the active ingredient to the diluent is 1:(1-10).

[0014] Furthermore, the volume ratio of the active ingredient to the diluent is 1:(5-10).

[0015] Furthermore, the diluent is liquid paraffin.

[0016] The fourth technical solution of the present invention:

[0017] The application of the *Tetrapanax papyrifer* attractant or the *Tetrapanax papyrifer* attractant preparation in the preparation of drugs that attract *Tetrapanax papyrifer*.

[0018] Furthermore, the application period is until mid-June.

[0019] Furthermore, when applying the *Tetrapanax papyrifer* attractant or the *Tetrapanax papyrifer* attractant preparation, it needs to be filled into a slow-release material and suspended in the cotton field.

[0020] Compared with the prior art, the present invention has the following advantages and technical effects:

[0021] The Chinese lacewing is a polyphagous dominant natural enemy that plays a role in pest control in various farmland ecosystems, while Cnidium monnieri is a functional plant that can attract a large number of natural enemies. This invention utilizes and applies the olfactory behavioral responses of the Chinese lacewing to volatile substances in Cnidium monnieri. The attractant obtained by this invention has the characteristics of significant attraction effect, harmlessness to other natural enemies, and environmental friendliness, providing a new reference method for biological control. Attached Figure Description

[0022] The accompanying drawings, which form part of this application, are used to provide a further understanding of this application. The illustrative embodiments and descriptions of this application are used to explain this application and do not constitute an undue limitation of this application. In the drawings:

[0023] Figure 1 The results of olfactory behavior induction of the attractant preparations for *Tetrapanax papyrifer* prepared in Examples 5-8;

[0024] Figure 2 The results of olfactory behavior induction of the attractant preparations for *Tetrapanax papyrifer* prepared in Comparative Examples 1-8 are shown. Detailed Implementation

[0025] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0026] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0027] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0028] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be readily apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0029] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0030] This invention provides an attractant for the Chinese lacewing, the active ingredient of which is composed of volatile substances from Cnidium monnieri, including (Z)-3-hexen-1-ol, γ-terpinene, trans-β-ocimene, and nerolidol.

[0031] The four volatile compounds (Z)-3-hexen-1-ol, γ-terpinene, trans-β-ocimene, and nerolidol used in the embodiments of this invention were all purchased from Beijing Coupling Technology Co., Ltd.

[0032] In this embodiment of the invention, the attractant for *Tetrapanax papyrifer* comprises, by volume, 10-15 parts of (Z)-3-hexen-1-ol, 10-20 parts of γ-terpinene, 60-70 parts of trans-β-ocimene, and 0.5-1 parts of nerolidol.

[0033] In a preferred embodiment of the present invention, the attractant for *Tetrapanax papyrifer* comprises, by volume, 12.9 parts of (Z)-3-hexen-1-ol, 18.8 parts of γ-terpinene, 67.6 parts of trans-β-ocimene, and 0.7 parts of nerolidol.

[0034] This invention also proposes a method for preparing the above-mentioned attractant for *Tetrapanax papyrifer*, by weighing (Z)-3-hexen-1-ol, γ-terpinene, trans-β-ocimene, and nerolidol by volume, and mixing them evenly.

[0035] This invention also proposes an attractant for *Tetrapanax papyrifer*, which consists of an active ingredient and a diluent. The active ingredient is the *Tetrapanax papyrifer* attractant, and the volume ratio of the active ingredient to the diluent is 1:(1-10).

[0036] In this embodiment of the invention, the volume ratio of the active ingredient to the diluent is 1:(5-10).

[0037] In this embodiment of the invention, the diluent is liquid paraffin, purchased from Shanghai Maclean Biochemical Technology Co., Ltd.

[0038] Unless otherwise specified, the term "parts" used in the embodiments of this invention refers to "volume parts".

[0039] The technical solution of the present invention will be further illustrated by the following embodiments.

[0040] Example 1

[0041] Weigh out 12.9 μL of (Z)-3-hexen-1-ol, 18.8 μL of γ-terpinene, 67.6 μL of trans-β-ocimene, and 0.7 μL of nerolidol, mix them thoroughly, and obtain the attractant for *Tetrapanax papyriferus*.

[0042] Measure 800 μL of liquid paraffin, add the prepared *Tetrapanax papyrifer* attractant to the liquid paraffin, and stir evenly to obtain the *Tetrapanax papyrifer* attractant preparation.

[0043] Example 2

[0044] Weigh out 15 μL of (Z)-3-hexen-1-ol, 20 μL of γ-terpinene, 60 μL of trans-β-ocimene, and 1 μL of nerolidol, mix them evenly, and obtain the attractant for *Tetrapanax papyriferus*.

[0045] Measure 768 μL of liquid paraffin, add the prepared *Tetrapanax papyrifer* attractant to the liquid paraffin, and stir evenly to obtain the *Tetrapanax papyrifer* attractant preparation.

[0046] Example 3

[0047] Weigh out 13 μL of (Z)-3-hexen-1-ol, 15 μL of γ-terpinene, 68 μL of trans-β-ocimene, and 0.85 μL of nerolidol, mix them thoroughly, and obtain the attractant for *Tetrapanax papyriferus*.

[0048] Measure 870 μL of liquid paraffin, add the prepared *Tetrapanax papyrifer* attractant to the liquid paraffin, and stir evenly to obtain the *Tetrapanax papyrifer* attractant preparation.

[0049] Example 4

[0050] Weigh out 12.9 μL of (Z)-3-hexen-1-ol, 18.8 μL of γ-terpinene, 68 μL of trans-β-ocimene, and 0.85 μL of nerolidol, mix them thoroughly, and obtain the attractant for *Tetrapanax papyriferus*.

[0051] Measure 603 μL of liquid paraffin, add the prepared *Tetrapanax papyrifer* attractant to the liquid paraffin, and stir evenly to obtain the *Tetrapanax papyrifer* attractant preparation.

[0052] Example 5

[0053] Weigh out 15 μL of (Z)-3-hexen-1-ol, 20 μL of γ-terpinene, 68 μL of trans-β-ocimene and 0.7 μL of nerolidol, mix them evenly to obtain the attractant for *Tetrapanax papyriferus*.

[0054] Measure 600 μL of liquid paraffin, add the prepared *Tetrapanax papyrifer* attractant to the liquid paraffin, and stir evenly to obtain the *Tetrapanax papyrifer* attractant preparation.

[0055] Example 6

[0056] Same as Example 5, except that 500 μL of liquid paraffin is measured.

[0057] Example 7

[0058] Same as Example 5, except that 1000 μL of liquid paraffin was measured.

[0059] Example 8

[0060] Same as Example 5, except that 800 μL of liquid paraffin was measured.

[0061] Example 9

[0062] Weigh out 10 μL of (Z)-3-hexen-1-ol, 10 μL of γ-terpinene, 70 μL of trans-β-ocimene, and 0.5 μL of nerolidol, mix them evenly, and obtain the attractant for *Tetrapanax papyriferus*.

[0063] Measure 90.5 μL of liquid paraffin, add the prepared *Tetrapanax papyrifer* attractant to the liquid paraffin, and stir evenly to obtain the *Tetrapanax papyrifer* attractant preparation.

[0064] Example 10

[0065] Same as Example 9, except that 452.5 μL of liquid paraffin was measured.

[0066] Comparative Example 1

[0067] Same as Example 1, except that (Z)-3-hexen-1-ol is replaced with an equal volume of γ-terpinene:

[0068] Weigh out 31.7 μL of γ-terpinene, 67.6 μL of trans-β-ocimene and 0.7 μL of nerolidol, mix them evenly to obtain the attractant for *Tetrapanax papyriferus*.

[0069] Measure 800 μL of liquid paraffin, add the prepared *Tetrapanax papyrifer* attractant to the liquid paraffin, and stir evenly to obtain the *Tetrapanax papyrifer* attractant preparation.

[0070] Comparative Example 2

[0071] Same as Example 1, except that γ-terpinene is replaced with an equal volume of (Z)-3-hexen-1-ol:

[0072] Weigh out 31.7 μL of (Z)-3-hexen-1-ol, 67.6 μL of trans-β-ocimene, and 0.7 μL of nerolidol, mix them thoroughly, and obtain the attractant for *Tetrapanax papyriferus*.

[0073] Measure 800 μL of liquid paraffin, add the prepared *Tetrapanax papyrifer* attractant to the liquid paraffin, and stir evenly to obtain the *Tetrapanax papyrifer* attractant preparation.

[0074] Comparative Example 3

[0075] Same as Example 1, except that trans-β-ocimene is replaced with an equal volume of nerolidol:

[0076] Weigh out 12.9 μL of (Z)-3-hexen-1-ol, 18.8 μL of γ-terpinene and 68.3 μL of nerolidol, mix them evenly and obtain the attractant for *Tetrapanax papyriferus*.

[0077] Measure 800 μL of liquid paraffin, add the prepared *Tetrapanax papyrifer* attractant to the liquid paraffin, and stir evenly to obtain the *Tetrapanax papyrifer* attractant preparation.

[0078] Comparative Example 4

[0079] Add 100 μL of (Z)-3-hexen-1-ol to 800 μL of liquid paraffin and stir until homogeneous.

[0080] Comparative Example 5

[0081] Add 100 μL of γ-terpinene to 800 μL of liquid paraffin and stir until homogeneous.

[0082] Comparative Example 6

[0083] Add 100 μL of trans-β-ocimene to 800 μL of liquid paraffin and stir until homogeneous.

[0084] Comparative Example 7

[0085] Add 100 μL of nerolidol to 800 μL of liquid paraffin and stir until homogeneous.

[0086] Comparative Example 8

[0087] Same as Example 1, except that 1500 μL of liquid paraffin was measured and the prepared *Tetrapanax papyrifer* attractant was added to the liquid paraffin.

[0088] Experimental Example 1: Olfactory Behavioral Method

[0089] The selective preference behavior of *Tetrapanax papyrifer* for the attractants prepared in all examples was observed using a Y-type olfactometer.

[0090] Place the Y-shaped olfactory probe on a table and use a photometer to measure the light intensity on both arms to ensure uniform illumination. Connect the sample bottle (blank), gas flow meter, distilled water humidifier, activated carbon filter, and air sampler sequentially to both ends of the Y-shaped olfactory probe using silicone tubing. The air sampler acts as a pump to dispense gas, the activated carbon filter reduces interference from pollutants, and the distilled water humidifier moistens the air.

[0091] Before the experiment, the adult natural enemy insects (*Corydalis sinensis*) were starved for 4 hours. The atmospheric sampler was then powered on and aerated for 10 minutes to fill the entire Y-shaped olfactory instrument. The gas flow meter was adjusted. During the experiment, the starved adult insects were introduced one by one into the main arm of the olfactory instrument. Their behavioral responses were observed. Insects that crawled beyond half the length of a odor source tube arm and remained there for more than 15 seconds were considered selected. Insects that did not reach half the length of a odor source tube arm within 5 minutes were considered not selected. After testing 5 insects, the inner and outer walls of the tube were wiped with 95% ethanol, dried, and the directions of the odor source and control source in the Y-shaped tube were reversed to eliminate errors. The sample bottle was set as a single-component *Cnidium monnieri* volatile compound (the *Corydalis sinensis* attractant prepared in Examples 5-8), with air entering from the bottom and exiting from the top, serving as the odor source. A control group was prepared by dripping liquid paraffin onto a 10×40mm paper strip. 30 male and 30 female adults were tested in each group.

[0092] The olfactory behavior of different experimental cases of *Chrysalis sinensis* was assessed using the chi-square test, and the differences between males and females were analyzed using the t-test. First, the raw data from lacewings with both arms were processed, and then the chi-square test and t-test were used for comparative analysis.

[0093] The olfactory behavior results of the attractants for *Tetrapanax papyrifer* prepared in Examples 5-8 are shown below. Figure 1 The horizontal axis represents the selection rate of ladybugs, that is, the ratio of the attractant preparation of *Tetrapanax papyrifer* prepared by selection 5-8 to the liquid paraffin of the control group (the same below). Example 5 refers to Example 5, and so on. ns indicates that there is no significant difference between the two (the same below), * indicates that there is a significant difference between the two, and ** indicates that there is a highly significant difference between the two. Figure 1 The olfactory behavior test results of Example 5 (female adult: χ 2 =9.60, P=0.005; Male adults: χ 2 =13.07, P=0.001), Example 6 (female adult: χ 2 =6.67, P=0.02; Male adults: χ² 2 =17.07, P<0.001), Example 7 (female adult: χ 2=5.40, P=0.02; Male adults: χ 2 =4.28; P=0.038), Example 8 (female adult: χ 2 =6.67, P=0.01; Female adult: χ 2 The attractant preparations for *Tetrapanax papyrifer* prepared with a concentration of 4.27 (P = 0.039) showed significant attraction for *Tetrapanax papyrifer*.

[0094] The attractants prepared in Comparative Examples 1-8 were determined using the same method described above, and the results are shown in the figure. Figure 2 . Figure 2 The olfactory behavior test results showed that, compared with the attractant proposed in this invention, the attractants prepared in Comparative Examples 1-8 were all more effective against *Tetrapanax papyrifer* (χ²) 2 (<0.001, P>0.05) No significant attraction effect.

[0095] Application examples

[0096] The attractants for *Tetrapanax papyrifer* prepared in Examples 1-8 were placed in a slow-release device (centrifuge tube, the cap of which was fitted with two small holes made of wire and filled with absorbent cotton but not soaked in the attractant). The slow-release device was attached to a whiteboard and hung in a cotton field in Shihezi City, Xinjiang. The attractant prepared in Comparative Example 1 was treated in the same way. Each treatment was repeated 5 times, with two films spaced between each treatment. The hanging height was approximately 1.3 meters, and the hanging time was in early August. The number of *Tetrapanax papyrifer* on the whiteboard was counted after 3 days, and the results are shown in Table 1.

[0097] Table 1. Attraction effect of *Tetranychus sinensis* in cotton fields in Shihezi City, Xinjiang in 2023.

[0098] Example 1 45 Example 2 37 Example 3 28 Example 4 31 Example 5 43 Example 6 31 Example 7 22 Example 8 30 Comparative Example 1 7

[0099] From Table 1 and Figure 1 The survey data shows that the attractant prepared in this invention has a significant attraction effect on the Chinese lacewing.

[0100] The above are merely preferred embodiments of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in this application should be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. A attractant for the Chinese lacewing, characterized in that, Its active ingredients consist of volatile substances from Cnidium monnieri, which, by volume, include: 10-15 parts of (Z)-3-hexen-1-ol, 10-20 parts of γ-terpinene, 60-70 parts of trans-β-ocimene, and 0.5-1 parts of nerolidol.

2. The attractant for *Tetrapanax papyriferus* according to claim 1, characterized in that, The composition by volume includes: 12.9 parts of (Z)-3-hexen-1-ol, 18.8 parts of γ-terpinene, 67.6 parts of trans-β-ocimene, and 0.7 parts of nerolidol.

3. A method for preparing the attractant for *Tetrapanax papyrifer* as described in any one of claims 1-2, characterized in that, Weigh out the (Z)-3-hexen-1-ol, γ-terpinene, trans-β-ocimene and nerolidol according to volume fractions, and mix them evenly to obtain the final product.

4. A type of attractant for the Chinese lacewing, characterized in that, It consists of an active ingredient and a diluent, wherein the active ingredient is the attractant for *Tetrapanax papyrifer* as described in any one of claims 1-2, and the volume ratio of the active ingredient to the diluent is 1:(1-10).

5. The attractant for *Tetrapanax papyriferus* according to claim 4, characterized in that, The volume ratio of the active ingredient to the diluent is 1:(5-10).

6. The attractant for *Tetrapanax papyrifer* according to claim 4, characterized in that, The diluent is liquid paraffin.

7. The use of the attractant for *Tetrapanax papyrifer* according to any one of claims 1-2 in the preparation of a drug for attracting *Tetrapanax papyrifer*.

8. The application according to claim 7, characterized in that, The application period is before mid-June.

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