Grapholitha molesta repellent as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of green prevention and control of pests, and particularly relates to a grapholitha molesta repellent as well as a preparation method and application thereof. The invention provides a grapholitha molesta repellent composed of volatile substances of mint and basil and a grapholitha molesta repellent preparation obtained by diluting the repellent, the grapholitha molesta repellent preparation is used for preventing and controlling grapholitha molesta, and the method is a method for repelling pests by using chemical information substances. The botanical repellent preparation or repellent prepared by the invention has the advantages of environmental friendliness, no residue, safety and no toxicity to human and livestock, and has a wide application prospect under the concept of green and sustainable development of agriculture at present.

Description

Technical Field

[0001] The present invention belongs to the technical field of green pest control, and more specifically relates to a Grapholita molesta repellent and its preparation method and application. Background Art

[0002] Grapholita molesta belongs to the family Tortricidae of the order Lepidoptera, also known as the peach shoot borer, etc., abbreviated as Grapholita molesta, and is a world-wide fruit-boring pest, widely distributed in Asia, Australia, Europe and North America. Grapholita molesta has a wide range of host plants, including peach, apple, pear, plum, prune, Chinese flowering crabapple, cherry and hawthorn, etc. Grapholita molesta has the harmful habit of transferring hosts, and mainly damages by the larvae boring into the tender shoots and fruits. In the early stage, it bores into the tender shoots of peach trees, forming a shoot-breaking shape, resulting in the withering of peach trees. In the later stage, it damages fruits such as pears, and can bore into the fruit core and discharge feces into the fruit. The damaged fruits often turn black and rot at the boring hole, commonly known as "black plaster". In recent years, with the adjustment of agricultural industrial structure and the large-scale concentrated planting of fruit trees, the damage of Grapholita molesta has become increasingly serious. In severe cases, it can even lead to the failure of the orchard, and the prevention and control situation is becoming increasingly severe. At present, the control of Grapholita molesta still mainly relies on spraying chemical pesticides. However, due to the characteristics of boring damage and overlapping generations of this pest, the chemical control effect is often not ideal, and the high-dose and high-frequency application of pesticides has also caused a series of problems such as environmental pollution, sharp reduction in the number of natural enemies, enhanced pest resistance and pesticide residues in fruits.

[0003] How to provide a repellent with the advantages of environmental friendliness, no residue, safety and non-toxicity to humans and livestock to control Grapholita molesta under the concept of green and sustainable agricultural development and replace highly toxic chemical pesticides has become the primary problem in the current research field of new plant protection technologies. Summary of the Invention

[0004] The purpose of the present invention is to provide a Grapholita molesta repellent and its preparation method and application to solve the problems existing in the above-mentioned prior art.

[0005] To achieve the above purpose, the present invention provides the following solutions:

[0006] One of the technical solutions of the present invention: Provide a Grapholita molesta repellent, and the active ingredient of the Grapholita molesta repellent is composed of volatile substances;

[0007] The volatile substances include: methyl conenone, ethyl 3-methylbutyrate, decanal and nonanal.

[0008] The volatile substances for repelling Grapholita molesta are composed of multiple components. Through screening and combination, the above-mentioned Grapholita molesta repellent is obtained in the present invention, and the dosage of each volatile substance directly determines the repellent effect of the Grapholita molesta repellent.

[0009] Further, the volatile substances are composed of methyl conenone, ethyl 3-methylbutyrate, decanal, and nonanal.

[0010] Further, by volume, the volatile substances include: 30 - 40 parts of methyl conenone, 30 - 40 parts of ethyl 3-methylbutyrate, 10 - 20 parts of decanal, and 10 - 20 parts of nonanal.

[0011] In the present invention, methyl conenone, ethyl 3-methylbutyrate, decanal, and nonanal are extracted from mint and basil by the headspace adsorption method. After the volatile substances are extracted, the specific components are identified through electroantennogram (EAD) and gas chromatography - mass spectrometry (GC - MS) analysis.

[0012] Optionally, by volume, the volatile substances include: 37 parts of methyl conenone, 37 parts of ethyl 3-methylbutyrate, 13 parts of decanal, and 13 parts of nonanal.

[0013] The second technical solution of the present invention: provides a preparation method of the above-mentioned Grapholita molesta repellent, and the steps include:

[0014] Mix methyl conenone, ethyl 3-methylbutyrate, decanal, and nonanal, and after stirring evenly, the Grapholita molesta repellent is obtained.

[0015] The third technical solution of the present invention: provides a Grapholita molesta repellent preparation, and the components include the above-mentioned Grapholita molesta repellent and a diluent.

[0016] Further, the Grapholita molesta repellent preparation is composed of the Grapholita molesta repellent and a diluent.

[0017] Further, the volume ratio of the Grapholita molesta repellent to the diluent is 1:1 - 10.

[0018] Optionally, the volume ratio of the Grapholita molesta repellent to the diluent is 1:5 - 10.

[0019] Further, the diluent is n-hexane.

[0020] The fourth technical solution of the present invention: provides a preparation method of the above-mentioned Grapholita molesta repellent preparation, and the steps include:

[0021] Mix the Grapholita molesta repellent with the diluent, and after stirring evenly, the Grapholita molesta repellent preparation is obtained.

[0022] Fifth technical solution of the present invention: Provide an application of the above-mentioned Grapholita molesta repellent or the above-mentioned Grapholita molesta avoidance preparation in controlling Grapholita molesta.

[0023] Sixth technical solution of the present invention: Provide an application of the above-mentioned Grapholita molesta repellent or the above-mentioned Grapholita molesta avoidance preparation in the preparation of drugs for controlling Grapholita molesta.

[0024] The present invention discloses the following technical effects:

[0025] The plant-derived avoidance preparation or repellent provided by the present invention belongs to the category of chemical ecological regulation and is composed of plant secondary metabolites and their derivatives. Through the identification of the components and structures of plant secondary metabolites and their derivatives, the mechanism of regulating insect behavior is clarified. Furthermore, these plant secondary metabolites with high regulatory effects are developed into avoidance preparations or repellent products, which can be used for pest control. The plant-derived avoidance preparation or repellent prepared by the present invention has the advantages of environmental friendliness, no residue, safety and non-toxicity to humans and livestock. Moreover, under the concept of current agricultural green and sustainable development, it has broad application prospects. It can not only replace highly toxic chemical pesticides to a certain extent, but also reduce the usage amount of chemical pesticides, and further solve problems such as pest resistance to pesticides, environmental pollution, and excessive agricultural residues.

[0026] The Grapholita molesta repellent or Grapholita molesta avoidance preparation prepared by the present invention can be hung in the orchard to repel Grapholita molesta and reduce its damage. It is suitable for popularization and application in the orchard and has good economic, social and ecological benefits and broad industrialization prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The drawings constituting a part of the present invention are used to provide a further understanding of the present invention. The schematic embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation to the present invention. In the drawings:

[0028] Figure 1 It is the olfactory avoidance effect of the avoidance preparations prepared in Examples 5-8 on Grapholita molesta adults.

[0029] Figure 2 It is the olfactory avoidance effect of the avoidance preparations prepared in Comparative Examples 1-7 on Grapholita molesta adults.

[0030] Figure 3 It is the Y-shaped insect olfactometer used in the test example. DETAILED DESCRIPTION OF THE INVENTION

[0031] Now, various exemplary embodiments of the present invention will be described in detail. This detailed description should not be considered as a limitation to the present invention, but should be understood as a more detailed description of certain aspects, characteristics and implementation schemes of the present invention.

[0032] It should be understood that the terms used in the present invention are only for describing specific embodiments and are not intended to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0033] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0034] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific embodiments of the description of the present invention, which are obvious to those skilled in the art. Other embodiments obtained from the description of the present invention are obvious to those skilled in the art. The description and examples of the present invention are merely exemplary.

[0035] Regarding "comprising", "including", "having", "containing", etc. used herein, they are all open-ended terms, meaning including but not limited to.

[0036] It should be noted that the aspects not described in detail in the present invention are all conventional operating means in the art and are not the focus of the present invention.

[0037] The raw materials and reagents involved in the specific embodiments of the present invention are all commercially available products without special instructions, and the purchase channels do not affect the realization of the technical effects.

[0038] The embodiments of the present invention provide a Grapholita molesta repellent, the active ingredient of which consists of the volatile substances of mint and basil, and the volatile substances include methyl conenone, ethyl 3-methylbutyrate, decanal, and nonanal.

[0039] The methyl conenone, ethyl 3-methylbutyrate, decanal, and nonanal used in the embodiments of the present invention are all purchased from Beijing Coupling Technology Co., Ltd., and are obtained by headspace adsorption method.

[0040] In some specific embodiments, the Grapholita molesta repellent comprises, by volume parts: 30 - 40 parts of methyl conenone, 30 - 40 parts of ethyl 3-methylbutyrate, 10 - 20 parts of decanal, and 10 - 20 parts of nonanal.

[0041] In some specific embodiments, the pear borer repellent comprises, by volume, 37 parts of methylconenone, 37 parts of ethyl 3-methylbutyrate, 13 parts of decanoic acid, and 13 parts of nonanal.

[0042] In a specific embodiment of the present invention, a method for preparing the above-mentioned pear borer repellent is also proposed, which is obtained by weighing the methylconenone, ethyl 3-methylbutyrate, aldehyde and nonanal according to the above-mentioned volume fractions, mixing and stirring evenly.

[0043] The specific embodiment of the present invention also provides a pear borer repellent preparation, which is composed of an active ingredient and a diluent. The active ingredient is the pear borer repellent, and the volume ratio of the active ingredient to the diluent is 1:1-10.

[0044] In some preferred embodiments of the present invention, the volume ratio of the active ingredient to the diluent is 1:5-10.

[0045] In a specific embodiment of the present invention, the diluent is n-hexane, which is purchased from Shanghai MacLean Biochemical Technology Co., Ltd.

[0046] Unless otherwise specified, the "parts" used in the embodiments of the present invention are parts by volume.

[0047] The technical solution of the present invention is further illustrated by the following embodiments.

[0048] Example 1

[0049] The preparation steps of the pear borer repellent include:

[0050] Mix methylconenone, ethyl 3-methylbutyrate, nopanal and nonanal in a volume ratio of 37:37:13:13 to obtain a pear borer repellent.

[0051] Example 2

[0052] The preparation steps of the pear borer repellent include:

[0053] Mix methylconenone, ethyl 3-methylbutyrate, nopanal and nonanal in a volume ratio of 35:35:15:15 to obtain a pear borer repellent.

[0054] Example 3

[0055] The preparation steps of the pear borer repellent include:

[0056] Methylconenone, ethyl 3-methylbutyrate, nopanal and nonanal were uniformly mixed in a volume ratio of 33:33:17:17 to obtain a pear borer repellent.

[0057] Example 4

[0058] The preparation steps of the Grapholita molesta repellent include:

[0059] Mix methyl conenone, ethyl 3-methylbutyrate, decanal, and nonanal evenly according to a volume ratio of 31:31:19:19 to obtain the Grapholita molesta repellent.

[0060] Example 5

[0061] The preparation steps of the Grapholita molesta repellent preparation include:

[0062] Mix the Grapholita molesta repellent prepared in Example 1 and n-hexane evenly according to a volume ratio of 1:6 to obtain the Grapholita molesta repellent preparation.

[0063] Example 6

[0064] Compared with Example 5, the difference is that the Grapholita molesta repellent prepared in Example 1 and n-hexane are mixed evenly according to a volume ratio of 1:5.

[0065] Example 7

[0066] Compared with Example 5, the difference is that the Grapholita molesta repellent prepared in Example 1 and n-hexane are mixed evenly according to a volume ratio of 1:7.

[0067] Example 8

[0068] Compared with Example 5, the difference is that the Grapholita molesta repellent prepared in Example 1 and n-hexane are mixed evenly according to a volume ratio of 1:9.

[0069] Comparative Example 1

[0070] Compared with Example 1, the only difference is that decanal is replaced with an equal volume of nonanal:

[0071] Weigh 37 μL of methyl conenone, 37 μL of ethyl 3-methylbutyrate, and 26 μL of nonanal, and mix them evenly to obtain the Grapholita molesta repellent.

[0072] Measure 900 μL of n-hexane, add the prepared Grapholita molesta repellent to the n-hexane, and stir evenly to obtain the Grapholita molesta repellent.

[0073] Comparative Example 2

[0074] Compared with Example 1, the only difference is that nonanal is replaced with an equal volume of decanal:

[0075] Weigh 37 μL of methyl conenone, 37 μL of ethyl 3-methylbutyrate, and 26 μL of decanal, and mix them evenly to obtain the Grapholita molesta repellent.

[0076] Measure 900 μL of n - hexane, add the prepared Grapholita molesta repellent to the n - hexane, and stir evenly to obtain the Grapholita molesta repellent.

[0077] Comparative Example 3

[0078] Compared with Example 1, the difference is only that methyl conenone is replaced with an equal volume of ethyl 3 - methylbutyrate:

[0079] Weigh 74 μL of ethyl 3 - methylbutyrate, 13 μL of nonanal, and 13 μL of decanal, and mix evenly to obtain the Grapholita molesta repellent.

[0080] Measure 900 μL of n - hexane, add the prepared Grapholita molesta repellent to the n - hexane, and stir evenly to obtain the Grapholita molesta repellent.

[0081] Comparative Example 4

[0082] Compared with Example 1, the difference is that methyl conenone is used as the active ingredient of the Grapholita molesta repellent. Specifically:

[0083] Add 100 μL of methyl conenone to 900 μL of n - hexane and stir evenly.

[0084] Comparative Example 5

[0085] Compared with Example 1, the difference is that ethyl 3 - methylbutyrate is used as the active ingredient of the Grapholita molesta repellent. Specifically:

[0086] Add 100 μL of ethyl 3 - methylbutyrate to 900 μL of n - hexane and stir evenly.

[0087] Comparative Example 6

[0088] Compared with Example 1, the difference is that decanal is used as the active ingredient of the Grapholita molesta repellent. Specifically:

[0089] Add 100 μL of decanal to 900 μL of n - hexane and stir evenly.

[0090] Comparative Example 7

[0091] Compared with Example 1, the difference is that nonanal is used as the active ingredient of the Grapholita molesta repellent. Specifically:

[0092] Add 100 μL of nonanal to 900 μL of n - hexane and stir evenly.

[0093] Test Example

[0094] The effects of the Grapholita molesta repellent preparations prepared in Examples 5-8 were detected. Specifically, a Y-tube olfactometer was used to observe the choice preference behavior of adult Grapholita molesta towards the Grapholita molesta repellent preparations prepared in Examples 5-8.

[0095] The Y-type insect olfactometer ([ Figure 3 ) was selected for the experiment. The Y-tube consisted of a glass tube with a diameter of 2.6 cm, which was equipped with a base and two arms, each 18 cm long, and the angle between them was 70°. Before the experiment, all glassware was cleaned with anhydrous ethanol and dried. 10 μL of the solution was dropped onto a rectangular filter paper (2 cm × 1 cm) and placed in the odor source bottle, and 10 μL of n-hexane control reagent was dropped onto the filter paper in the other odor bottle. The instruments used were connected in the order of an air sampler (QC-1S, Beijing Institute of Labor Protection Science), a gas flow meter, an activated carbon filter (250 mL), a distillation flask humidifier, an odor source device, and a Y-tube using a silica gel tube. The power was turned on, and the air sampler was observed and adjusted to make the air flow rates on both sides the same (250 mL / min, LZB-3WB, Changzhou Kede Thermal Instrument Co., Ltd.). Finally, a 20W table lamp (light intensity 2000 lx) was placed on the side of the two arms. Female (male) adult Grapholita molesta were introduced into the main arm of the Y-tube and observed for 5 min. When the adult climbed to 1 / 3 of the side wall and stayed for more than 5 seconds, it was recorded as having a tropism; otherwise, there was no response. The left and right arm directions of the Y-tube were changed every 5 Grapholita molesta tested, and the Y-tube was changed every 10 tested. Each Grapholita molesta was only used for one test, and a new filter paper was replaced for each Grapholita molesta tested. Each group was repeated 30 times, and all tests were carried out from 19:00 to 24:00.

[0096] The olfactory selection results of female and male adult Grapholita molesta for different test examples were analyzed by chi-square test. First, the original data of the number of Grapholita molesta selecting the two arms were weighted, and then the chi-square test in non-parametric tests was used for comparative analysis.

[0097] The olfactory results are as Figure 1 shown, Figure 1 which is the olfactory repellent effect of the repellent preparations prepared in Examples 5-8 on adult Grapholita molesta. In the figure, the horizontal axis represents the selection rate of Grapholita molesta for different treatments, specifically the selection rate of Grapholita molesta for the Grapholita molesta repellent preparations prepared in Examples 5-8 and the control n-hexane. ns indicates that there is no significant difference between the two (the same below), * represents that there is a significant difference between the two, and ** represents that there is an extremely significant difference between the two. Figure 1 The olfactory behavior test results showed that in Example 5 (female adults: χ 2 = 7.00, P = 0.008; male adults: χ 2= 5.538, P = 0.019), Example 6 (female adults: χ 2 = 7.00, P = 0.008; male adults: χ 2 = 4.481, P = 0.034), Example 7 (female adults: χ 2 = 8.333, P = 0.004; male adults: χ 2 = 14.286; P < 0.001), Example 8 (female adults: χ 2 = 6.259, P = 0.012; female adults: χ 2 = 8.333, P = 0.004) The Grapholita molesta repellent preparation prepared has a significant repellent effect on Grapholita molesta.

[0098] The repellent preparations prepared in Comparative Examples 1-7 were measured by the same method as above, and the results are shown in Figure 2 . Figure 2 The olfactory behavior test results of show that compared with the repellent preparation proposed in the present invention, the Grapholita molesta repellent preparations prepared in Comparative Examples 1-7 have no significant repellent effect on Grapholita molesta (P > 0.05).

[0099] Application Example

[0100] The Grapholita molesta repellent or repellent preparation prepared in Examples 1-8 was placed in a slow-release device (centrifuge tube, with two small holes pierced in its lid with iron wire, and cotton wool placed without immersing the repellent), and the slow-release device was hung in the apple orchard at 30 / mu. The repellent prepared in Comparative Example 1 was treated in the same way. Each treatment had 3 replicates. The hanging time was at the beginning of June, and the slow-release device was replaced once a month (the amount of the repellent or repellent preparation placed in the slow-release device tube was excessive, that is, there was still remaining after 1 month of volatilization). The investigation was carried out at the end of August. 100 apples were randomly investigated for each treatment to calculate the fruit borer rate, and the results are shown in Table 1.

[0101] Table 1 Effects of different treatments on the fruit borer rate of Grapholita molesta

[0102] Sample Fruit borer rate (%) Example 1 10.33±2.08a Example 2 12.67±3.05ab Example 3 11.67±3.21ab Example 4 13.33±3.06ab Example 5 15.33±2.51ab Example 6 13.67±2.52ab Example 7 17.00±2.00b Example 8 15.33±3.06ab Comparative Example 1 29.00±4.36c

[0103] Note: Different letters represent significant differences at the 0.05 level.

[0104] From Table 1 and Figure 1 the data, it can be seen that the repellent preparation and repellent preparation prepared in the present invention have a significant repellent effect on Grapholita molesta.

[0105] In this specification, each example is described in a progressive manner. The key point of each example is to illustrate the differences from other examples. The same and similar parts among the examples can be referred to each other.

[0106] The foregoing description of the disclosed embodiments enables those skilled in the art to practice or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A Grapholita molesta repellent, characterized in that, The active ingredient of the Grapholita molesta repellent consists of volatile substances; The volatile substances include: methyl conenone, ethyl 3-methylbutyrate, decanal, and nonanal.

2. The Grapholita molesta repellent according to claim 1, wherein By volume fraction, the volatile substances include: 30-40 parts of methyl conenone, 30-40 parts of ethyl 3-methylbutyrate, 10-20 parts of decanal, and 10-20 parts of nonanal.

3. The Grapholita molesta repellent according to claim 2, characterized in that, By volume fraction, the volatile substances include: 37 parts of methyl conenone, 37 parts of ethyl 3-methylbutyrate, 13 parts of decanal, and 13 parts of nonanal.

4. A method for preparing the Grapholita molesta repellent according to any one of claims 1 to 3, characterized in that the steps Include: Mix methyl conenone, ethyl 3-methylbutyrate, decanal, and nonanal, and obtain the Grapholita molesta repellent after stirring evenly.

5. A Grapholita molesta repellent preparation, characterized in that, The components include the Grapholita molesta repellent according to any one of claims 1-3 and a diluent.

6. The Grapholita molesta repellent preparation according to claim 5, characterized in that, The volume ratio of the Grapholita molesta repellent to the diluent is 1:1-10; and / or, the diluent is n-hexane.

7. A preparation method of the Grapholita molesta repellent preparation according to claim 6, characterized in that the steps Include: Mix the Grapholita molesta repellent with the diluent, and obtain the Grapholita molesta repellent preparation after stirring evenly.

8. Use of the Grapholita molesta repellent according to any one of claims 1-3 or the Grapholita molesta repellent preparation according to claim 6 in controlling Grapholita molesta.

9. Use of the Grapholita molesta repellent according to any one of claims 1-3 or the Grapholita molesta repellent preparation according to claim 6 in preparing a drug for controlling Grapholita molesta.

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