阅读说明：本技术 一种趋避草地贪夜蛾的吴茱萸提取物及其应用 (Evodia rutaecarpa extract for evading spodoptera frugiperda and application thereof ) 是由 唐照磊 苏月贵 杜青 李石初 于 2021-02-05 设计创作，主要内容包括：本发明公开了一种趋避草地贪夜蛾的吴茱萸提取物及其应用,制剂来源于药用植物吴茱萸果实及叶片的提取物,对玉米草地贪夜蛾具有长期趋避作用,减少草地贪夜蛾在玉米植株上的产卵,达到长效防治草地贪夜蛾的目的,节省了劳动力,保护了农田生态环境,为研究开发草地贪夜蛾趋避剂奠定基础及提供原料。(The invention discloses a evodia rutaecarpa extract for repelling Spodoptera frugiperda and application thereof, wherein a preparation is derived from extracts of medicinal plant evodia rutaecarpa fruits and leaves, has a long-term repelling effect on corn Spodoptera frugiperda, reduces spawning of Spodoptera frugiperda on corn plants, achieves the purpose of long-acting control of Spodoptera frugiperda, saves labor force, protects the ecological environment of a farmland, lays a foundation for researching and developing Spodoptera frugiperda repelling agents and provides raw materials.)

1. The fructus evodiae extract for avoiding spodoptera frugiperda is characterized by comprising the following components in parts by weight: the volatile gas comprises the following components in percentage by weight:

2. the method for preparing evodia rutaecarpa extract for evading spodoptera frugiperda as claimed in claim 1, wherein the method comprises the following steps: the method comprises the following steps:

1) pulverizing fructus evodiae fruit and leaf;

2) weighing the crushed fructus evodiae fruits and leaves, putting the crushed fructus evodiae fruits and leaves into a beaker, adding cyclohexanone and tert-butyl alcohol according to the mass ratio of 1:2.5:0.5, and soaking and extracting for 8-24 h;

3) filtering the extractive solution, and evaporating cyclohexanone to obtain fructus evodiae extract.

3. The method for preparing evodia rutaecarpa extract for evading spodoptera frugiperda according to claim 2, wherein the method comprises the following steps: the soaking extraction in the step 2) lasts for 24 hours.

4. The method for preparing evodia rutaecarpa extract for evading spodoptera frugiperda according to claim 2, wherein the method comprises the following steps: and 3) evaporating cyclohexanone by using a rotary evaporator at the conditions of 60 ℃ and 150 rpm.

5. Use of the evodia rutaecarpa extract for repelling spodoptera frugiperda of any one of claims 1 to 4 for the preparation of a repellent for controlling spodoptera frugiperda.

[ technical field ] A method for producing a semiconductor device

The invention relates to the technical field of plant-derived pesticides, and particularly relates to an evodia rutaecarpa extract for evading spodoptera frugiperda and application thereof.

[ background of the invention ]

Spodoptera frugiperda (J.E. Smith) belongs to Lepidoptera (Lepidotera) and Spodoptera (Noctuidae) and also called fall armyworm, originates from the American area, is a major international migratory pest for global early warning of grain and agriculture organizations in United nations, has the characteristics of wide suitable living area, strong migratory flight capability, high propagation multiple, heavy overeating hazard, high control difficulty and the like, rapidly rolls over a plurality of provinces and cities in China since 1 month in 2019, and brings great threat to food production in China.

The lawn spodoptera frugiperda can be propagated and survived at the proper temperature of 11-30 ℃, and can not live through winter in the northern areas of China with the average daily temperature close to or lower than 9 ℃, but because the imagoes can migrate and have excellent capacity, the imagoes can migrate for 100-500 km in the air flow of the northern areas of China in summer and one night, and the imagoes can reach 1600km farthest, so the imagoes are endowed with a different name of the armyworm. The temperature rises in spring every year, and the imagoes migrate to the north. The growth speed of Spodoptera frugiperda becomes faster along with the rise of air temperature, and the Spodoptera frugiperda can be propagated for several generations in one year. The Spodoptera frugiperda breeding generation only needs 30 days in summer, 60 days in spring and autumn and 80-90 days in winter. The service life of the adult is as long as 15-20d, more than 1500 eggs can be laid once, and the adult can be hatched after 2-4 d. Spodoptera frugiperda belongs to omnivorous pests, larvae like eating tender leaves, can bite a large amount of gramineae such as rice, sugarcane, cotton, peanut, soybean and corn and other grain crops of 80 kinds of crops of Compositae, Brassicaceae and the like, and high-age larvae can bite roots and stems of the crops and even penetrate into corn ears; if the crop is found to be untimely, the crops can be completely damaged for several acres at night, the loss is generally not equal to 20% -60%, and serious field or even no crop is received, so that serious economic loss is caused.

Depending on the type of the pest crop, spodoptera frugiperda can be classified into two major types, corn (mainly pest corn, cotton, etc.) and rice (mainly pest rice, alfalfa, etc.). At present, spodoptera frugiperda which is a nuisance in China is mainly of a corn type. According to statistics, Spodoptera frugiperda occurs in more than 1500 counties (cities and districts) with 26 provinces in China in 2019, the corn is damaged by Spodoptera frugiperda in China in about 3000 mu in area in the south province of China in 2020, and huge loss is brought to local corn production. The pest can cause damage to leaves, ears, male flowers, filaments and other parts of the corn in the whole growth period. The seedling is damaged in the period from the staminate stage to cause blade perforation, and the plant is withered after the growing point is gnawed; male flowers are damaged to cause that the corn plants can not bloom; the maize can not be pollinated after the filament is damaged; after the ears are damaged, the yield of grains is reduced, and the corn ear rot is caused to occur greatly. If the common areas do not carry out pesticide control, the yield is reduced by more than 50 percent and even the area is completely harvested.

The Spodoptera frugiperda has the characteristics of migration, serious overlapping, sudden increase of food consumption after four-instar larvae, explosive property and the like, most of the conventional insecticides can effectively control the pests, but because the migration and egg laying amount of adults are large, and the insecticides generally only have a lasting period of about 7-10 days, the insecticides are required to be used for controlling for many times in one production cycle. Investigation finds that Guangxi in 2020 is used as a frequent region of Spodoptera frugiperda, farmers need to use 3-7 times of chemical pesticide to control Spodoptera frugiperda in each corn production period, so that the investment of labor force and production cost is greatly increased, and the problems of food safety, environmental pollution and the like are caused, so that the exploration of a long-acting, green, low-cost and low-labor-investment control medicament and a control method for controlling Spodoptera frugiperda is an important current task.

At present, chemical pesticides are still the main means for controlling spodoptera frugiperda. According to statistics, on corn, 5% of seedlings have stem breakage, and 20% of young plant leaf cluster (30 days before growth) is damaged, and chemical control is generally needed. On sorghum, the economic threshold for this pest is 1 (or 2) larvae per leaf, or 2 on each ear. However, because the spodoptera frugiperda larvae are mostly hidden in the recurrent leaves of plants such as corn and the like, the larvae are difficult to directly reach by spraying chemical pesticide; in addition, Spodoptera frugiperda has now developed strong resistance to chemical insecticides such as organophosphates, carbamates and pyrethroids. Meanwhile, the use of chemical pesticides in large quantities also creates a lot of ecological environmental problems, for example, the enrichment of chemical pesticides kills a lot of beneficial insects, i.e. bees and the like. In view of this, it is very important to control Spodoptera frugiperda by using a biological control method.

Along with the increasing awareness of people in advocating nature, protecting environment and paying attention to food safety, the pollution-free biopesticide industry and the biological control research field have acquired an inexhaustible development opportunity. The species of natural enemy insects of spodoptera frugiperda are found to be more, and parasitic wasps, parasitic flies and predatory stinkbugs are mainly found. These natural enemy insects achieve better control effects in some areas by parasitizing or preying on spodoptera frugiperda larvae.

The plant-derived pesticides of spodoptera frugiperda are found in many types, and are mainly effective in matrine, rotenone, azadirachtin emulsion and the like. Research shows that matrine has lethal and sub-lethal effects on Spodoptera frugiperda, but the lethality rate is relatively low. The study of domestic scholars shows that after 0.3% of matrine aqueous solution, 6% of rotenone microemulsion and 0.3% of azadirachtin emulsifiable solution are used for treating spodoptera frugiperda for 5 days at the temperature of 20 ℃, the humidity of 70% and the illumination of 16/8 hours, the lethality rates are 41%, 20% and 5% respectively. Therefore, in order to achieve a better effect of controlling spodoptera frugiperda, the plant-derived pesticide needs to be further optimized in components.

The research of medicinal plants as biological pesticides is reported in China, for example, Chinese patent application CN201610132440, a preparation method of evodia rutaecarpa as agricultural fungicide and its application are prepared by extracting evodia rutaecarpa fruit, fruit stalk, fruit shell and leaf; wherein the weight percentage of the main active components is as follows: 6-10% of hydroxyevodiamine, 5-10% of evodiamine, 5-10% of rutaecarpine, 6-10% of dehydroevodiamine, 1-5% of evodiamine, 1-5% of rutaecarpine and 6-40% of other alkaloid components; the plant extract is processed into aqueous emulsion, and can be used for preventing and treating various plant diseases after being diluted. For example, in chinese patent application CN202010141543, an application of fructus forsythiae extract for controlling agricultural pests is prepared by placing fructus forsythiae and folium forsythiae in a constant temperature oven at 75 deg.c, drying to constant weight, pulverizing, sieving, adding 70% ethanol at a ratio of 1: 10-20(g/mL) respectively, reflux-extracting at 78-82 deg.c for 2-4h, extracting for three times, filtering, mixing filtrates, rotary evaporating the filtrate, and concentrating to obtain fructus forsythiae and folium forsythiae ethanol extract for controlling agricultural pests including armyworm and corn borer.

Although the use of biogenic pesticides for preventing and controlling Spodoptera frugiperda is reported at home and abroad, reports and related patents that volatile matters of fructus evodiae fruits and leaves have an evasive effect on the Spodoptera frugiperda imagoes are not retrieved.

[ summary of the invention ]

The invention aims to overcome the defects of food safety, environmental pollution and labor waste caused by the fact that an insecticide is required to be used for multiple times in the existing corn spodoptera frugiperda prevention and control technology, and provides an evodia rutaecarpa extract for avoiding spodoptera frugiperda and application thereof.

The purpose of the invention is realized by the following technical scheme:

the evodia rutaecarpa extract for avoiding spodoptera frugiperda comprises the following volatile gases in percentage by weight:

the invention also aims to provide a preparation method of the evodia rutaecarpa extract for avoiding Spodoptera frugiperda, in particular to a preparation method of the evodia rutaecarpa fruit and leaf extract for avoiding Spodoptera frugiperda, which comprises the following steps:

1) pulverizing fructus evodiae fruit and leaf;

2) weighing the crushed fructus evodiae fruits and leaves, putting the crushed fructus evodiae fruits and leaves into a beaker, adding cyclohexanone and tert-butyl alcohol according to the mass ratio of 1:2.5:0.5, and soaking and extracting for 8-24 h;

3) filtering the extractive solution, and evaporating cyclohexanone to obtain fructus evodiae extract.

In the invention:

the soaking extraction in the step 2) lasts for 24 hours.

And 3) evaporating cyclohexanone by using a rotary evaporator at the conditions of 60 ℃ and 150 rpm.

The invention also aims to provide application of the evodia rutaecarpa extract for repelling spodoptera frugiperda in preparation of a repellent for preventing and treating spodoptera frugiperda.

Compared with the prior art, the invention has the following advantages:

1. compared with the common method, the preparation method of the evodia rutaecarpa extract for avoiding spodoptera frugiperda in the invention adds cyclohexanone and tert-butyl alcohol, and mainly aims to extract vinyl components such as beta-Myrcene (beta. -Myrcene), D-Limonene (D-Limonene), beta-Phellandrene (beta. -Phellandrene), beta-ocimene (1,3,6-Octatriene,3,7-dimethyl-, (Z) -) and has good effect on preventing and treating spodoptera frugiperda.

2. The evodia rutaecarpa extract for avoiding spodoptera frugiperda is used for preventing and treating spodoptera frugiperda by utilizing volatile matters of evodia rutaecarpa fruit and leaf extracts, and the characteristic that the smell sense of pests is sensitive to some substances and is far away from the avoided substances is utilized, so that compared with the traditional chemical pesticide, the evodia rutaecarpa extract has the advantages of environmental friendliness, long lasting period, easiness in operation and the like; compared with the traditional biological insecticide, the field control operation is easier to implement, and a large amount of labor force is not needed for spraying. The evodia rutaecarpa extract is determined by the imago evasion, the influence on the egg laying habit of the spodoptera frugiperda imagoes in a greenhouse and field control experiments, so that the evodia rutaecarpa extract has a strong evasion effect on the spodoptera frugiperda, can be used as a good spodoptera frugiperda imago evasion agent in vegetable fields, and has important significance for advocating green control of plant diseases and insect pests.

[ description of the drawings ]

FIG. 1 is a positive ion mode diagram of a Total Ion Current (TIC) diagram of a sample of volatile gas of fructus evodiae extract obtained in example 1 of the present invention, which is detected by a gas chromatography-mass spectrometry combination.

FIG. 2 is a negative ion pattern diagram of Total Ion Current (TIC) of a sample of volatile gas of fructus evodiae extract obtained in example 1 of the present invention by combined gas chromatography-mass spectrometry.

[ detailed description ] embodiments

The following examples are provided to further illustrate the embodiments of the present invention.

Example 1:

a preparation method of an evodia rutaecarpa extract for evading spodoptera frugiperda comprises the following steps:

1) crushing fructus evodiae fruit and leaves by using a wall breaking machine;

2) weighing the crushed fructus evodiae fruits and leaves, putting the crushed fructus evodiae fruits and leaves into a beaker, adding cyclohexanone and tert-butyl alcohol according to the mass and volume of 1:2.5:0.5, and soaking and extracting for 24 hours;

3) filtering the extractive solution, and evaporating cyclohexanone at 60 deg.C and 150rpm with rotary evaporator to obtain fructus evodiae extract.

The components and the content of the obtained evodia rutaecarpa extract are shown in table 1 after volatile gas of the extract is detected by adopting a gas chromatography-mass spectrometry combination method, wherein the extract is obtained from evodia rutaecarpa fruit and leaf extract and has the activity of avoiding the spodoptera frugiperda of corn:

TABLE 1 volatile ingredients and contents of Evodia rutaecarpa extract

Example 2:

a preparation method of an evodia rutaecarpa extract for evading spodoptera frugiperda comprises the following steps:

1) crushing the fruit and the leaves of the evodia rutaecarpa by a crusher;

2) weighing the crushed fructus evodiae fruits and leaves, putting the crushed fructus evodiae fruits and leaves into a beaker, adding cyclohexanone and tert-butyl alcohol according to the mass and volume of 1:2.5:0.5, and soaking and extracting for 8 hours;

3) filtering the extractive solution, and evaporating cyclohexanone at 60 deg.C and 150rpm with rotary evaporator to obtain fructus evodiae extract.

Example 3:

a preparation method of an evodia rutaecarpa extract for evading spodoptera frugiperda comprises the following steps:

1) crushing fructus evodiae fruit and leaves by using a wall breaking machine;

2) weighing the crushed fructus evodiae fruits and leaves, putting the crushed fructus evodiae fruits and leaves into a beaker, adding cyclohexanone and tert-butyl alcohol according to the mass and volume of 1:2.5:0.5, and soaking and extracting for 16 hours;

3) filtering the extractive solution, and evaporating cyclohexanone at 60 deg.C and 150rpm with rotary evaporator to obtain fructus evodiae extract.

Comparative example 1:

the preparation process is carried out in step 2) with cyclohexanone in a mass/volume ratio of 1:3, as in example 1.

Comparative example 2:

the preparation method in step 2) is carried out by adding methanol according to the mass to volume ratio of 1:3, and the method is otherwise the same as that in example 1.

Comparative example 3:

the preparation method comprises the step 2) of adding cyclohexanone and methanol according to the mass: volume ratio of 1:2.5:0.5, and the steps are the same as those of the example 1.

As a result:

TABLE 2 partial volatile components and contents of evodia rutaecarpa extract obtained in examples and comparative examples

The results show that:

compared with the comparative example, in the embodiment of the invention, after the crushed fructus evodiae fruits and leaves are weighed, cyclohexanone and tert-butyl alcohol are added according to the mass: volume ratio of 1:2.5:0.5, and soaking and extraction are carried out, so that alkene components such as beta-Myrcene (beta. -Myrcene), D-Limonene (D-Limonene), beta-Phellandrene (beta. -Phellandrene), beta-ocimene (1,3,6-Octatriene,3,7-dimethyl-, (Z) -) can be extracted to the maximum extent, and the method has a good effect on subsequent use for preventing and controlling spodoptera frugiperda.

Experimental example:

1. indoor determination of repelling and avoiding effect of extract on Spodoptera frugiperda imagoes

The extracts obtained in example 1 and comparative example 1 are used as raw materials, an insect olfaction tester is used for measuring the evasion of volatile matters to Spodoptera frugiperda adults, the brand and model of a Y-shaped insect olfaction tester is TianlingJSTL 3-150, the product of Jiangsu Tianlinginstrumental company Limited is used, the ventilation volume is 0.16pb during testing, and the time length of each measurement is 2 min. The total number of measurements was 3. The avoidance rate is calculated according to the following formula: the avoidance rate (%) is (number of population in CK zone-number of population in treated zone) × 100/(number of population in CK zone + number of population in treated zone).

TABLE 2 Spodoptera frugiperda reaction on volatiles of Chinese herbal extracts in a Y-type insect olfaction tester

The results show that:

the number of adults was 1 on average near the air inlet through which the volatile extract of evodia rutaecarpa of example 1 was introduced, the number of adults was 8.3 on average near the air inlet through which the volatile extract of evodia rutaecarpa of comparative example 1 was introduced, and the number of adults was 16 on average near the air inlet through which clean air (CK) was introduced, and it was comprehensively calculated that the avoidance rate of the volatile extract of evodia rutaecarpa of example 1 to adults of spodoptera frugiperda was 94.12%, and the avoidance rate of the volatile extract of evodia rutaecarpa of comparative example 1 to adults of spodoptera fru.

2. Example 1 Net Room assay of the avoidance Effect of Evodia rutaecarpa extract on adults of Spodoptera frugiperda

After the fructus evodiae extract of example 1 was dissolved in acetone at a ratio of 1:1, sterile water was added to dilute the solution to 500-fold and 800-fold solutions of 1000-fold, and 1 drop of tween 80 was added to prepare a solution for future use, as a control, an equal concentration of acetone + tween 80 solution.

The specification of the net room is that length is multiplied by width and multiplied by height is 6m multiplied by 2m multiplied by 2.2m, 4 cells are arranged in the net room, each cell is 1.5 m, corn is sown in the cell, the sowing specification is that row spacing is 50cm multiplied by plant spacing 20cm, row length is 2m, 30 corn seedlings are reserved in each cell, and after the seedlings grow to 4 leaves, 4 cells are respectively correspondingly treated by 4 types: spraying 500 times, 800 times and 1000 times of medicinal liquid of fructus evodiae extract and control solution, randomly arranging each treatment, placing 20 female Spodoptera frugiperda and 10 male Spodoptera frugiperda in the net room, and investigating egg laying amount in each cell treatment after 5 days. The experiment was set up in 3 replicates (3 net rooms). The avoidance rate is calculated according to the following formula: the results of the avoidance rate (%) (number of CK-treated egg pieces) × 100/(number of CK-treated egg pieces + number of treated egg pieces) are shown in table 3 below:

TABLE 3 number of eggs laid by Spodoptera frugiperda after spraying of corn leaf with Evodia extracts of various concentrations

The results show that:

after the corn leaves are sprayed with 500-fold, 800-fold and 1000-fold solutions of the evodia rutaecarpa extract in example 1, all of the spodoptera frugiperda oviposits are concentrated on the leaves of the control group plants which are not sprayed with the spodoptera frugiperda, the evodia rutaecarpa extract has an avoidance effect on spodoptera frugiperda, and the avoidance rates of the 500-fold, 800-fold and 1000-fold diluted solutions on the spodoptera frugiperda yield are all 100%;

the aversion rates of 500-fold, 800-fold and 1000-fold diluted solutions of the evodia rutaecarpa extract of comparative example 1 to the spodoptera frugiperda production were 22.22%, 46.67% and 62.96%, respectively;

the aversion rates of 500-fold, 800-fold and 1000-fold diluted solutions of the evodia rutaecarpa extract of comparative example 2 to the spodoptera frugiperda production were 29.03%, 42.86% and 53.85%, respectively;

the 500-fold, 800-fold and 1000-fold diluted solutions of the evodia rutaecarpa extract of comparative example 3 had aversion rates to spodoptera frugiperda production of 20.00%, 54.84% and 65.52%, respectively.

The results show that the avoidance effect of the example on the oviposition of Spodoptera frugiperda is obviously higher than that of the comparative examples 1 to 3 under the same concentration compared with the comparative examples 1 to 3.

3. Control test in the field

3.1 preparation of the drug delivery device: a plastic toothpick box (the diameter is 4.5cm, the height is 8cm) is punched with 6 small holes around the position, which is about 3cm away from a cover, of the box wall by a pair of red tweezers, a group of absorbent cotton is placed in the box, the absorbent cotton occupies about one half of the volume of the box, one end of a No. 8 iron wire is spirally bent around the box to form a spiral supporting body, the box is fixed, the other end of the iron wire is bent and folded to form a triangular support, the triangular support is convenient to insert underground and serves as a fixture, and the height of the box and the ground is ensured to be about 60 cm.

3.2 Experimental design and implementation: the test area is 10.3 mu land, the land is rectangular, the length is 122m, the width is 56m, a natural isolation zone with the circumference of about 200 m is arranged on the periphery of the test area, the isolation zone is a waste greenhouse, trees with the length of about 5-7m naturally grow in the greenhouse, the distance between the control area and the test area is 280 m, and corns (the variety is Guidan 162) are sowed at the same time. In order to determine that spodoptera frugiperda occurs in both the control area and the test area, no pesticide is used after the corn seedlings emerge, after the spodoptera frugiperda occurs and the pest rate reaches about 30%, 5% of emamectin benzoate and hexaflumuron (trade name is Yingzui) is used for spraying once on the test area and the control area simultaneously, the harmful larvae and ova are eliminated, meanwhile, the extract in the example 1 is used for preventing and treating in the test area, and the pesticide is not used in the control area again for preventing and treating.

The method of use of the extract of example 1 was: the application devices prepared in 3.1 are placed at intervals of 12m on the periphery of the land, 30ml of extract is added into a box of each application device, absorbent cotton in the box is enabled to fully absorb the liquid medicine, and the extract in the example 1 is supplemented every 7 days.

The investigation method comprises the following steps: and (3) respectively adopting a five-point sampling method to investigate the filament damage rate in a control area and a test area in the maize milk stage, wherein each point is taken for 2 rows, 50 maize plants are continuously investigated in each row, the control effect is totally investigated according to the filament damage rate, and the control effect (%) (the average damage rate of the filaments in the control area-the average damage rate of the filaments in the treatment area) is 100/the average damage rate of the filaments in the control area.

3.3 results

The test area and the control area are sprayed with 5% emamectin benzoate-hexaflumuron (trade name is Yingzui) once in 9-13-month of 2020, the same day of the test area is sprayed with 800 times of the solution of the extract in example 1 for aversion control, and the growth period of the corn is V7. Corn filament damage was investigated at 28 days 10 months.

TABLE 4 field control effect of evodia rutaecarpa extract for evading control of Spodoptera frugiperda

The results show that: the average damage rate of the filaments in the maize milk stage in the test area is 4.8 percent, the damage rate of the filaments in the maize milk stage in the control area is 93.8 percent, and the field control effect of the extract in the example 1 for avoiding and controlling spodoptera frugiperda is 94.88 percent.

The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and variations can be made without departing from the inventive concept of the present invention, which falls into the protection scope of the present invention.