CN114304185B - Locust extract and application thereof - Google Patents
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- CN114304185B CN114304185B CN202011064002.9A CN202011064002A CN114304185B CN 114304185 B CN114304185 B CN 114304185B CN 202011064002 A CN202011064002 A CN 202011064002A CN 114304185 B CN114304185 B CN 114304185B
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Abstract
The invention belongs to the technical field of pesticides, and particularly relates to a locust extract and application thereof in serving as an anti-locust preparation. The extract is obtained by extracting locust and/or ovum thereof. The locust extract is used as an anti-locust agent. The locust extract and the active substances in the extract are extracted from the locust itself as a raw material, and are further used for the self-resistance treatment of the locust; the anti-locust agent does not contain any additive or exogenous compound, has good anti-locust effect, and is safe and nontoxic to the environment.
Description
Technical Field
The invention belongs to the technical field of pesticides, and particularly relates to a locust extract and application thereof in serving as an anti-locust preparation.
Background
Arthropoda is the largest animal species in the world, known as arthropods, and about 120 tens of thousands of species exist worldwide, accounting for 80% of the total number of species present. Arthropods living in extremely wide environments, whether sea water, fresh water, soil, and air, have their traces.
Insects, which are arthropods of the phylum Arthropoda, are diverse in variety and morphology, are the most numerous groups of animals on earth, and their trails are distributed almost throughout every corner of the world. Insects are the most abundant species in the animal kingdom, and the greatest number of insects has a significant impact on agricultural production and human health. Insects are generally classified into the orders orthoptera, the order of the tail of the bullet, the order of the thysanoptera, the order of the coleoptera and the order of the hemiptera, with locust, butterfly, aphid, fly, mosquito, dragonfly, cockroach, etc. being most common. The current world is more severely affected by insect disasters concentrated on grasshopper.
The migratory locusts, belonging to phylum arthropoda, class of entomophyta, order orthoptera, including grasshopper general, class of grasshopper general, about 14000 species worldwide, more than 1200 species in our country, more than 60 species in severe harm, among which, east asia migratory locusts (Locusta migratoria manilensis Meyen), asia migratory locusts (Locusta migratoria migratoria l.), tibetan migratory locusts (Locusta migratoria tibetensis Chen) 3 migratory sub-species, and desert migratory locusts (Schistocerca gregaria Forsk), asia trolley locusts (Oedaleus asiaticus Bei-Bienko), italian locusts (Calliptamus italicus l.), siberia locusts (Gomphocerus sibiricus l.), etc., can form off-site sudden disasters. Among them, the east Asia migratory locust has the most extensive distribution in China and the most serious harm, and is the most main migratory locust species causing the disaster of China.
The propagation speed of the migratory locust in east Asia is high, the propagation amount is large, the survival period is long, 2 to 4 generations can occur each year, and the average number of spawning of female adult locusts per generation can be 300 to 400. The food has wide feeding range and large feeding capacity, the number of the food plants reaches more than 20, and 40-70 kg of crops can be eaten every ten thousands of adult locusts every day, so that 30-50% of yield is reduced, and even the yield is prevented. The migratory grasshopper is divided into a colonial type and a discrete type, the former is appeared in the big occurrence year of the high-density grasshopper group, the latter is present in the small occurrence year when the density is very low, and the two types are still in transition type (or intermediate type) and are formed by dispersing the high-density the nymph of a locust group or aggregating the low-density the nymph of a locust group. The locusts in the high-density locusts have bunching property, and after the bunching, the locusts are manic and have rich aggressiveness, the appetite is greatly increased, the locusts can migrate into groups for a long time and long distance without eating or drinking, and the great loss can be caused to agriculture.
Historically, the world suffered serious locusts, 1875, and the united states experienced the most terrorist locusts in the history of humans, 12.5 trillions of locusts attack the western united states, causing tremendous losses. In the taro-polo region of south russia in 2001, grasslands and cultivated lands of about 26.7 ten thousand hectares are affected by grasshoppers. In 2004, millions of locusts hit israel erlatt, destroying a large number of crops and flowers, and israel governments have used aircraft to spray pesticides to kill locusts. In 2020, the feeble of african locusts, egyptian, somali, kennia, etc., was attacked by hundreds of millions of locusts; the Egypt-Russian government was first to announce national emergency entry and to issue an famine warning. While China is one of countries suffering serious locust disaster damage all the time, and the damage of the locust disaster always aims at agriculture in China from spring and autumn to dateBringing about a great loss. According to the records of spring and autumn, 12 locust plagues occur in the Shandong province; chinese book records that grasses and trees are gnawed to be empty when the grasses are covered by the grasses and the grasses are covered by the grasses; chinese locust records that the disaster of the migratory locust causes more than seven hundred thousand of disaster people in sixteen years of China. In the early establishment period of new China, the generation area of the migratory locust in east Asia is about 521 ten thousand hm 2 . In the 80 s, migratory locust in east Asia frequently occurs in Huang-Huai-Hai region and southwest of Hainan island, and the annual occurrence area is about 100 to 150 ten thousand hm 2 Agricultural production is severely compromised. In autumn in 1985, 10 tens of thousands of acres of reed leaves and hundreds of acres of corn ears She Chiguang are taken off from the south to the Hebei farm after the high-density populated grasshopper group of the migratory grasshopper in the east Asia in Tianjin, northern harbor, and the swept area reaches 250 tens of thousands of acres. A large number of migratory locust disasters in China, the disaster causing core area of the migratory locust disasters is 80 ten thousand hm in the places such as Hebei, henan and Shandong and the like and the West and south extension thereof, 1998 and 1999 in two consecutive years 2 The above. The locust disaster area in the Xinjiang area in 2000 is 3480 tens of thousands mu, the locust disaster occurs in the North China in 2002, the Tianjin is serious, and the number of locust individuals in 1 square meter reaches 4000-5000. Locust plague occurs in the Shandong region in 2017, and crops are completely eaten, so that huge losses are brought to farmers. Therefore, locust control has important significance.
At present, biological control and chemical control are mainly adopted for locusts so as to control the occurrence and diffusion of locusts. The biological control is mainly to use biological agents, and controls the harm of the locust by protecting and utilizing the natural enemies of the locust or releasing the artificially cultivated parasitic natural enemies of the locust, such as increasing the number of natural enemies of chickens, ducks, birds and the like, or applying the microsporidians, the metarhizium anisopliae and the like of the locust with extremely strong killing power; although the biological control is friendly to the environment, the current biological control has the defects of changing the community structure, high control cost and immature control method. The chemical prevention and control usually adopts organophosphorus, pyrethrin and compound pesticides thereof, which play a positive role in rapidly controlling the occurrence and diffusion of locusts, but the chemical prevention and control is greatly limited because a large amount of pesticides can cause poisoning of people and livestock, environmental pollution and killing of other organisms and have the defect of poor safety. With the improvement of the environmental awareness of human beings, the search of novel locust resistant products which are safe, efficient, low in toxicity and environment-friendly in the nature has become a research hotspot. The existing researches show that certain components in the ethanol extracts of the buddleia macrophylla and the laggera pterodonta, the extracts of the Chinese azalea flower, the macleaya cordata fruit, the tripterygium wilfordii, the euphorbia, the sophora alopecuroide and the sparrow have certain poisoning activity on the locusts, but the extracts have poisoning activity on the locusts and other bodies or have poisoning activity on the locusts, and the components of active substances of the active locusts and the actual application thereof are still to be further studied.
Meanwhile, US6306415A1 in the prior art discloses an insect repellent comprising, as active ingredients, caprylic acid, pelargonic acid and capric acid in volatile silicone oils according to 1:1:1, proved to be capable of repelling flies. CN105899071a discloses mixtures of octyl, nonyl and decyl (C8, 9, 10) fatty acids with pyrethroids and also discloses methods of controlling aedes aegypti using mixtures of octyl, nonyl and decyl fatty acids with pyrethroids, wherein the mixtures of octyl, nonyl and decyl fatty acids with pyrethroids produce highly effective insecticidal compounds against adult mosquitoes and the insect repellents, octyl, nonyl and decyl fatty acids enhance the killing rate of the pyrethroids. Although the two documents disclose the use of caprylic acid, pelargonic acid and capric acid as insect repellent agents and the synergistic mosquito killing use of caprylic, pelargonic and capric fatty acids and pyrethroids; it only achieves the expelling of insects and no study of locust resistance with caprylic acid and/or capric acid alone, nor the relevant implications.
Disclosure of Invention
The invention aims at solving the problems existing in the prior art and provides a locust extract and application thereof as an anti-locust preparation.
In order to achieve the above purpose, the invention adopts the technical scheme that:
A locust extract is prepared from locust itself and/or its ovum as raw materials.
The locust is all locust of Arthropoda, class insect, order orthoptera and family of locust.
The locust is all of locust (according to Xia Kailing Charpy classification system) of Lauraceae, octomy, equidae, lepidaceae, equidae, and Equidae.
The Momordica Charantia sub-family includes Momordica Charantia sub-family, sophora Fusca sub-family, echinohtopinae and Prothetinae.
The Momordica charantia subfamily comprises the genus Tribulus, and the Tribulus genus comprises the Tribulus and the Tribulus inner Mongolicus.
The sub-family Umbelliferae comprises Umbelliferae the genus short nasal locust the genus Kepika, the genus Pseudonasosinusitis, the genus Tapix Yan Huang the genus of Protopetas Yan Huang, cirsium, bei Huang, mongolian, and silly (Umbelliferae).
The Ocimaceae comprises genus Huang Xinghuang, genus Ocimum, genus Olive-like locust, genus Yunnan locust, genus Mega, genus Vanilla, genus Canada.
The Uvulariaceae comprises Uvulariaceae, uvulariaceae.
The Philippine locust family comprises Momordica, clouds, momordica, momord the genus locusta, the genus locusta the genus of Alternaria, ulva Du Huang genus, taro genus, beatifying genus, glehnia genus, oryza genus, pseudo-oryza genus, cotton genus, desert genus (such as desert locust), spot leg locust genus, etc.
The Tripterygidae comprises genus of locustae, equisqualis genus the genus of locusts, migratory locusts, grass green locusts, spiked locusts, and the like the genus Octopus, the genus palustris, the genus Trogopterus, the genus humpback, the genus Alctopus Alternaria, and Alternaria, etc.
The Ulmaceae comprises Ulmaria, lepidoptera, ulmaria, sinkiang, kazakhstan, and grassland.
The Umbelliferae comprises Umbelliferae, embelliferae, roxburgh, and Mixarotea.
The Sword horn locust comprises long abdomen locust genus, desorptive field locust genus, chrysanthemum locust genus, summer locust genus, sword horn locust genus, etc.
The locusts are preferably locusts in east Asia, tibet, huang Jizhu, rice, sugarcane, winged, short-star-wing, benzene, negative, cotton, negative bamboo, siberian, halberd, dolly, pasture, young, scab, italian etc.
The extract is a substance formed by taking locust as a raw material, and directionally acquiring and/or concentrating one or more target components (namely components with locust resistance) in the raw material through a physicochemical extraction and separation process according to the requirements of locust resistance application of the extracted final substance. Including those obtained by a combination of extraction methods known in the art; the extraction method includes, but is not limited to, solvent treatment (water, ethanol, methanol, isopropanol, petroleum ether, benzene, chloroform, diethyl ether, ethyl acetate, dichloroethane, acetone, etc., oil, etc.), solid phase extraction, supercritical CO 2 Thermal, ultrasonic extraction, pressure or centrifugal force, or other chemical and/or mechanical processes to produce a target component from a feedstock, etc. Thus, the term "extract" as used herein includes crude extract, active ingredient and active monomer.
The "crude extract" in the present invention refers to a mixture of a target component with a lower concentration and a non-target component with a higher concentration obtained by extracting and separating a raw material with a solvent (water, ethanol, methanol, isopropanol, petroleum ether, benzene, chloroform, diethyl ether, ethyl acetate, dichloroethane, acetone, etc.).
The term "locust-resistant" as used herein refers to a reduction in the population and/or mobility of the locust, including not only killing of larvae and adults of the locust, but also prevention of development or hatching of eggs of the locust, inhibition of maturation, and inhibition of locust movement.
The method for killing the locust larvae and adults refers to the method that the bodies lose the metabolism capacity by a certain means; the 'preventing development or incubation of the locust eggs and inhibiting maturation' refers to a state that the hatching rate of the locust eggs is low and deviates from the normal development process (including high rate of teratogenesis caused by nymphs or adults in the development process and/or loss of reproductive capacity of the adults) by a certain means; the 'disabling teratogenesis' refers to that scattered wings, incomplete wing development or leg deformity of the locust occur in the development process from nymphs to adults; the expression "inhibiting the activity of the locust" means that the metabolism and/or activity ability of the locust is reduced by some means, including reduction, destruction, necrosis, etc. of the body of the locust.
The extract contains fatty acid compound, benzene compound, nitrogen compound or amino acid compound and one or more of the salts or esters of the above compounds (the above compounds are fatty acid compound, benzene compound, nitrogen compound or amino acid compound).
A locust extract is prepared from locust and/or ovum by defatting, extracting with organic solvent, concentrating extractive solution, and extracting with at least one organic solvent.
The extraction method can be one or more of squeezing method, leaching method, supercritical extraction method, water extraction method and solvent extraction method.
The organic solvent is one or more of alcohol ethanol, methanol, isopropanol, petroleum ether, chloroform, diethyl ether, ethyl acetate, dichloroethane and acetone; ethanol, methanol, isopropanol are preferred.
Degreasing raw material locust by using an organic solvent; the organic solvent may be petroleum ether, diethyl ether or n-hexane.
The organic solvent used in the extraction is methanol, ethanol, n-butanol, isopropanol, ethyl acetate, petroleum ether or acetone.
Further, the extraction method comprises the steps of degreasing locust with petroleum ether, extracting with ethanol, concentrating the extracting solution, extracting with at least one of petroleum ether, ethyl acetate and n-butanol in sequence, and concentrating.
An application of locust extract as anti-locust agent is provided.
The extract, the use of the active ingredient or the individual active monomers obtained by further processing of the extract as anti-locust agent.
The anti-locust substance is one or more of an extract obtained by extracting the locust and/or the eggs thereof as raw materials, an active component of the extract after further treatment, an active monomer in the active component of the extract after further treatment or an analogue of the active monomer in the active component of the extract after further treatment.
The active component in the extract contains one or more of fatty acid compounds, benzene compounds, nitrogen compounds or amino acid compounds and salts or esters of the compounds (the compounds are fatty acid compounds, benzene compounds, nitrogen compounds or amino acid compounds).
The fatty acid compound is a compound which consists of three elements of carbon, hydrogen and oxygen, has carboxyl on a carbon chain and can have hydroxyl on the carbon chain; wherein the carbon chain may be saturated or unsaturated.
The "salt of the aforementioned compound" according to the present invention is a salt in which hydrogen in a hydroxyl group or a carboxyl group in an active compound is substituted with a metal ion to form a corresponding salt. Further, the hydroxyl group or hydrogen in the carboxyl group in the active compound is replaced by a metal ion such as Na, K, etc. to form the corresponding salt.
The term "esters of the aforementioned compounds" according to the invention is the esterification of the carboxyl groups of the active compounds with alcohols to form the corresponding products.
The "active component" refers to an extract (relative to a crude extract) containing a target component at a higher concentration obtained by subjecting the crude extract to further treatments such as extraction, chromatography, distillation, sublimation, crystallization and precipitation, ion exchange, chromatographic separation, centrifugal separation, electrodialysis, electrolytic separation, electromagnetic separation, adsorptive separation, magnetic separation, and the like.
The "active monomer" refers to a target compound (i.e., an active compound having an anti-locust effect) in the above "active component".
The active component is one or more of fatty acid compounds, dibasic acid compounds, hydroxy fatty acid compounds, ester compounds, benzene-containing compounds, nitrogen-containing compounds or amino acid compounds and salts or esters of the compounds.
The fatty acid compound is one or more of (9Z, 11E) -conjugated linoleic acid, oleic acid, stearic acid, alpha-linolenic acid, linoleic acid, 14 (Z) -eicosenoic acid, 11, 14-eicosdienoic acid, myristic acid, lauric acid, n-pentadecanoic acid, decanoic acid, gluconic acid, n-octanoic acid, arachidonic acid, cork acid, wood wax acid and jasmonic acid, or salts or esters of the above substances; preferably cork acid, myristic acid, lauric acid, capric acid, n-caprylic acid or one or more of the salts or esters of the foregoing.
The dibasic acid compound is one or more of 3-tertiary butyl adipic acid, suberic acid, azelaic acid or salts or esters of the above substances.
The hydroxy fatty acid compound contains one or more of (15Z) -9,12, 13-trihydroxy-15-octadecenoic acid, 10-hydroxy-decenoic acid and cis-8, 11, 14-eicosatrienoic acid methyl ester.
The ester compound contains cis-8, 11, 14-eicosatrienoic acid methyl ester and ethyl myristate.
The amino acid compound contains DL-leucine.
The benzene-containing compound contains one or more of 4-indolecarboxaldehyde, 1-methyl-3, 6-pyrazinedione, benzoic acid, 2, 5-dihydroxybenzaldehyde, vanilloid, 4-cresol, salicylic acid, benzoic acid, gentisic acid, 3-phenoxypropionic acid, 4-aminobenzoic acid, o-chlorobenzoic acid or salts or esters of the above substances; preferably benzoic acid, vanilla, salicylic acid, gentisic acid or one or more of the salts or esters of the foregoing.
The nitrogen-containing compound contains one or more of 2- (acetamido) caproic acid, N-acetyl-L-phenylalanine, 7-methyladenine, acetaminophen, adenine, adenosine, 1-methyl-3, 6- (1H, 2H) piperazine dione or salts or esters of the above substances; preferably 2- (acetamido) hexanoic acid, N-acetyl-L-phenylalanine, acetaminophen, or one or more of the salts or esters of the foregoing.
An anti-locust substance, which is one of oleic acid, alpha-linolenic acid, linoleic acid, (9Z, 11 e) -conjugated linoleic acid, stearic acid, myristic acid, xylonic acid, 11, 14-eicosadienoic acid, 14 (Z) -eicosenoic acid, cork acid, jasmonic acid, lauric acid, N-pentadecanoic acid, decanoic acid, azelaic acid, 3-tert-butyladipic acid, suberic acid, (15Z) -9,12, 13-trihydroxy-15-octadecenoic acid, N-octanoic acid, gluconic acid, 10-hydroxy-decenoic acid, ethyl myristate, cis-8, 11, 14-eicosatrienoic acid methyl ester, salicylic acid, 4-indolecarboxaldehyde, 2, 5-dihydroxybenzaldehyde, arachidonic acid, 1-methyl-3, 6-pyrazinedione, benzoic acid, gentisic acid, 3-phenoxypropionic acid, 4-aminobenzoic acid, o-chlorobenzoic acid, adenine, adenosine, DL-leucine, vanilloid, 4-cresol, 2- (acetamido) caproic acid, N-acetyl-L-phenylalanine, 7-methyladenine, p-acetyl-1, 6-dimethyl-1 h-1, 6-diethyl ketone or 1 h; namely, one or more of the substances shown in Table 1 or Table 2; it can be obtained by further processing in the extract, commercially available, or synthesized in a conventional manner.
An anti-locust agent contains one or more of above anti-locust active ingredients. Namely, one or more of the extracts obtained by extracting the locusts and/or the eggs thereof as raw materials, the active components after the further treatment of the extracts, the active monomers in the active components after the further treatment of the extracts or the analogues of the active monomers in the active components after the further treatment of the extracts; or one or more of the active material monomer materials.
The locust resistant agent is prepared by mixing an active ingredient with an agriculturally acceptable carrier; wherein the active ingredient accounts for 0.01-99% of the weight of the locust resisting agent. Preferably 1 to 10%, more preferably 1%.
The use of said anti-locust agent as a protection for plants against locust stress; in particular as an application for protecting plants against the stress of migratory grasses in east Asia.
The locust-resistant composition comprises a component A and a component B, wherein the component A and the component B are mixed according to the weight part ratio of 1:100-100:1; the component A is one or more of the above locust-resistant substances; the component B is other compounds and/or pesticides with insecticidal, nematicidal, acaricidal, molluscicidal, bactericidal, virucidal, chemical sterilization and mating disruption properties, such as methamidophos, methiphos, fenpropaphos, monobromophos, nitenpyram, omethoate, parathion, ethiprole, fipronil, acrylonitrile, allethrin, D-cis-trans-allethrin, D-trans-allethrin, bifenthrin, bio-allethrin, abamectin, emamectin, phenazoquin, fenpyroximate, pyriminostrobin, pyridaben, fipronil, flumequine, benzimidazole, thiabendazole, cyprodinil and the like.
An anti-locust composition preparation, which is prepared by mixing the anti-locust composition as an active ingredient with an agriculturally acceptable carrier; wherein the active ingredient accounts for 0.01-99% of the weight of the locust-resistant composition. Preferably 1%.
The locust-resistant composition preparation may further comprise one or more of the following additives: surfactants, stabilizers, sustained release agents, defoamers, dispersants, wetting agents, disintegrants, binders, fillers, and the like.
The anti-locust agent and the anti-locust composition formulation can be in solid form and in liquid form.
The solid forms include, but are not limited to, powder, dust or granular formulations.
Such liquid formulation forms include, but are not limited to, dispersions, suspensions, emulsions, or solutions in a liquid carrier.
The preparation forms of the locust resisting agent and the locust resisting composition preparation can be powder, spraying agent, fumigating agent, coating agent and nanocrystallization preparation.
The powder may be applied to the soil; the spray is typically a solution, aqueous suspension or emulsion that can be applied to soil, plants or other areas that are stressed by grasses; the fumigant has a relatively high vapor pressure, can exist in gaseous form, and kills locusts in soil or enclosed spaces at sufficient concentrations; the nano preparation can be prepared by adopting a wet method medium grinding method, and the nano particle size of the nano preparation is 50-500nm, preferably 300-500nm.
A method of combating damage caused by a locust, the method comprising applying one or more of an extract obtained from the extraction of the locust itself as a starting material, an active substance in the extract thereof, an analogue of an active substance in the extract thereof or said formulation to any area which may be occupied, colonized and/or passed by a locust, thereby causing a reduction in the population and/or mobility of the locust (including not only killing the locust larvae and adults, but also preventing the development or hatching of the locust eggs, inhibiting maturation, and inhibiting the activity of the locust). Areas such as: areas of crop, tree, fruit, cereal, feed, vine, grass and/or ornamental plant growth; the area where humans or animals live and the soil surrounding the area.
The invention has the beneficial effects that:
(1) The locust extract and the active substances in the extract are extracted from the locust itself as a raw material, and are further used for the self-resistance treatment of the locust; the anti-locust agent does not contain any additive or exogenous compound, has good anti-locust effect, and is safe and nontoxic to the environment.
(2) The extracted locust extract and active substances in the extract can be used for surrounding plant materials subjected to locust stress by spraying, soaking, brushing and other methods.
(3) The crude extract and the active compound obtained by further processing after extraction are used as anti-locust products or active ingredients of the products act on the locusts, so that the locusts nymphs and adults can be killed within 1h, the hatching activity of locusts eggs is obviously reduced, the harm of the locusts is effectively resisted, the crude extract can be extracted through the locusts, and meanwhile, the compounds with the anti-locust effect are easy to obtain and can be purchased in the market; meanwhile, the active compound obtained by further processing the extract and the extract can be combined with other compounds with anti/insecticidal action, anti/locust killers, anti/insecticides, anti/acaricides, anti/bactericides and the like for use, and the product is favorable for subsequent marketing popularization and application.
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
The technical solutions of the embodiments of the present invention will be clearly and completely described below, and the described embodiments are only a part of the present invention, not the whole invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1 obtaining and chemical analysis of the extract of migratory locust
Main experimental material medicament: the egg mass of the migratory locust in east Asia, the adult migratory locust in east Asia (obtained by incubating and culturing the egg mass of the migratory locust in Emotion of Anhui province), petroleum ether, 85% ethanol, ethyl acetate and n-butanol.
Obtaining the locusta migratoria eggs: fresh migratory locust egg blocks are selected, and sediment is removed by rinsing with clear water.
Selecting adult locusts in east Asia: selecting adult locusts in east Asia with normal development and vitality.
Obtaining the extract of the locusta migratoria eggs: extracting oil from egg blocks of the migratory locust by petroleum ether (volume ratio petroleum ether: adult migratory locust=1:3), extracting at 75deg.C for 2h, and reflux extracting defatted egg blocks of the migratory locust with 85% ethanol (volume ratio defatted egg blocks of the migratory locust: 85% ethanol=1:10) for 2h after residual petroleum ether is volatilized; obtaining an extracting solution, and concentrating by rotary evaporation to remove a solvent, namely the extract of the locusta migratoria eggs; sequentially extracting the concentrated solution with petroleum ether, ethyl acetate and n-butanol with equal volumes, and concentrating the petroleum ether phase, ethyl acetate phase and n-butanol phase by rotary evaporation respectively to obtain petroleum ether liquid extract, ethyl acetate liquid extract and n-butanol liquid extract.
Obtaining an adult locusta migratoria extract: extracting crushed adult migratory locust grease (volume ratio petroleum ether: adult migratory locust = 1:3) with petroleum ether, extracting at 75deg.C for 2h, and reflux extracting defatted adult migratory locust with 85% ethanol (volume ratio defatted adult migratory locust: 85% ethanol = 1:10) for 2h after residual petroleum ether is volatilized; obtaining an extracting solution, and concentrating by rotary evaporation to remove a solvent, namely an adult locusta migratoria extract; sequentially extracting the concentrated solution with petroleum ether, ethyl acetate and n-butanol with equal volume, and concentrating the petroleum ether phase, ethyl acetate phase and n-butanol phase by rotary evaporation respectively to obtain petroleum ether liquid extract, ethyl acetate liquid extract and n-butanol liquid extract.
And (3) respectively carrying out component analysis on the obtained locusta migratory ovum and the obtained adult extract by using LC-MS to obtain the active components in the extract. The analysis conditions were as follows:
instrument: thermo Scientific Vanquish the ultra-high performance liquid chromatography system comprises a binary high pressure gradient pump, an automatic sampler, a column incubator, a vacuum degasser, and is coupled with a thermo scientific Q Exactive Orbitrap mass spectrometer for mass spectrometry, and the Q-exact host system comprises an H-ESI II source, an automatic syringe pump and a switching valve. The data was analyzed by Thermo Scientific Compound Discoverer software. The chromatographic column was a Water Atlantis T3 column (2.1 mm. Times.150 mm,3 μm)
LC/MS analysis: the chromatographic mobile phase was acetonitrile, 0.1% (v/v) formic acid in water. Acetonitrile and acid water gradient elution, the flow rate is 0.3ml/min, the elution time is 40min, the column temperature is 25 ℃, the sample injection amount is 3 mu L, the double electrospray ionization source works in a positive and negative ion mode to obtain a full-scanning mass spectrum of 50-1200m/z, the scanning rate is 1.0 spectrum/s, the source air temperature is set to 350 ℃, the additional source parameters are set to 40 and 15 of sheath and auxiliary gas flow respectively, and the scavenging flow is 0; the spraying voltage is 3.0kV; the capillary temperature was 325 ℃; the S-lens rf level is 60V. Data were collected by Masshanter Data Acquisition V B.04 and analyzed by Thermo Scientific Compound Discoverer software and the results of the analysis are shown in Table 1.
TABLE 1 LC/MS/MS analysis summary of the extracts of the egg of the migratory locust
TABLE 2 LC/MS/MS analysis summary of adult locusts in east Asia
Results: as can be seen from tables 1 and 2, the extracts of the eggs and adults of migratory locust in east Asia mainly comprise fatty acids, benzene rings, nitrogen-containing compounds and analogues thereof.
Example 2 incubation inhibition of the extract of Equipped migratory locust on its eggs and experiments for killing different age of locusts
1. Experiment of the adult Equipped with the extract of Equipped with the egg hatching inhibition
Fresh locusts are taken, the outer membrane of the locusts is peeled off, and locusts are reserved.
The petroleum ether extract, ethyl acetate extract and n-butanol extract of the adult migratory locust are respectively prepared into 1%, 5% and 10% solutions by water, and meanwhile, a clear water treatment group and a blank control group are taken for a control experiment. Pouring the treatment liquid into culture dishes respectively, putting 300 egg grains into each culture dish, and soaking for 10min; the eggs of the blank group were directly hatched without any treatment.
Placing the soaked egg particles and the treatment liquid into a locust egg hatching device, and then placing the hatching device into an artificial climatic chamber with RH of 50% and 39 ℃ and without illumination for culture. The hatchability of the daily locusts was recorded.
TABLE 3 statistical Table of the effects of adult extract of migratory locust on the hatching rate of the egg of the migratory locust
Note that: transferring the hatched nymphs, and carrying out other experimental purposes.
From the table, 10% petroleum ether extract, 5% ethyl acetate extract and 10% ethyl acetate extract have an inhibitory effect on the hatching of the locusts, wherein the concentration of the locusts treated by the 10% petroleum ether extract is the lowest, the hatching rate is 35.3%, and the locusts have a relatively excellent inhibitory effect on the hatching of the locusts.
2. Experiment for killing adult locusts in east Asia
And (3) selecting 1-instar larvae, 3-instar nymphs and 5-instar adults of the locusts hatched in the same day in a blank control group, spraying the treatment group solutions in the step (1), putting the nymphs into a feeding device every 30, repeating the steps every 5 times, replacing fresh corn leaves and clear water every day for feeding, and counting the growth conditions of the locusts in each group, wherein the counting results after 7 days are shown in tables 4-6.
TABLE 4 statistical Table of killing Rate of adult Equivocas locusts extract on 1 st instar larvae of Equivocas locusts (%)
TABLE 5 killing Rate statistics of adult Etoenail extract for 3-year-old nymphs of Etoenail (%)
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TABLE 6 statistical Table of the killing Rate of adult Etoena locusts extract on 5-year-old adult Etoena locusts (%)
As seen from tables 4 to 6, the petroleum extract and ethyl acetate extract of adult migratory locust in east Asia at concentrations of 5% and 10% had the killing activity against both 1-year-old and 3-year-old adults and adults, and in the concentration range of 1% -10%, the greater the concentration of the treatment liquid, the greater the killing activity.
3. Experiment of Equipped egg extract for suppressing hatching of Equipped egg
Fresh locusts are taken, the outer membrane of the locusts is peeled off, and locusts are reserved.
The extract of the migratory locust and the extract of the pebble oil ether, the extract of the ethyl acetate and the extract of the n-butanol are respectively prepared into solutions of 1 percent, 5 percent and 10 percent by water, and meanwhile, a clear water treatment group and a blank control group are taken for a control experiment. Pouring the treatment liquid into culture dishes respectively, putting 300 egg grains into each culture dish, and soaking for 10min; the eggs of the blank group were directly hatched without any treatment.
Pouring the soaked egg particles and the treatment liquid into a locust egg hatching device, and then placing the hatching device into an artificial climate box with RH of 50% and 39 ℃ and without illumination for culture. The hatchability of the daily locusts was recorded.
TABLE 7 statistical Table of the effect of Equipped locusta migratory egg extract on the hatchability of Equipped locusta eggs (%)
Note that: transferring the hatched nymphs, and carrying out other experimental purposes.
From the table, the concentration of 10% of petroleum ether extract and 10% of ethyl acetate extract has an inhibitory effect on the hatching of the locusts, wherein the hatching rate of the locusts in east asia after the treatment of the petroleum ether extract with the concentration of 10% is the lowest, the hatching rate is 45.2%, and the relatively better effect of inhibiting the hatching of the locusts is achieved.
4. Experiment of killing Equipped with egg extract of Equipped with different ages
And selecting 1-instar larvae, 3-instar nymphs and 5-instar adults of the locusts hatched in the same day in a blank control group, spraying the solutions of the treatment groups in the step 3, putting the nymphs into a feeding device every 30, repeating the steps every 30, replacing fresh corn leaves and clear water every day for feeding, and counting the growth conditions of the locusts in each group, wherein the counting results after 7 days are shown in tables 8-10.
TABLE 8 statistical Table of killing Rate of Equivoca locusts egg extract on 1 st instar larvae of Equivoca locusts (%)
TABLE 9 killing Rate statistics of Ocimum gratissimum egg extract for 3-year-old nymphs of Ocimum gratissimum (%)
TABLE 10 killing Rate statistics of Equipped locusts egg extract for 5 th year adults (%)
As seen from tables 8 to 10, the toenail extract and the ethyl acetate extract of the migratory locust with concentrations of 5% and 10% have the killing activity against the 1-year, 3-year and adult insects, and in the concentration range of 1% to 10%, the greater the concentration of the treatment liquid, the stronger the killing activity.
EXAMPLE 3 experiments on hatching inhibition of Octonoic acid on Equivoca migratoria eggs and killing of different-age locusts
1. Hatching inhibition experiment of octanoic acid on locusta migratoria eggs
And (3) preparation of a medicament: octanoic acid (purchased from market) is prepared into octanoic acid solutions with different concentrations through a solvent system; the solvent system comprises absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system.
Experimental group:
1) 10mL of octanoic acid solution with the concentration of 10 g/L;
2) 10mL of octanoic acid solution with the concentration of 1 g/L;
control group: the solvent control group (the solvent system is absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system) and the blank experiment are used as the control group.
Egg grain treatment: and taking fresh locusts in east Asia, peeling off the outer membrane of the locusts, only leaving locusts in east Asia, separating the locusts one by one, and ensuring the integrity of the locusts without mechanical damage to the locusts in the whole process.
And (3) medicament treatment: pouring the prepared treatment group solution and solvent treatment solution into culture dishes respectively, then placing fresh migratory locust egg grains, placing 300 egg grains into each culture dish, and soaking for 10min; the eggs of the blank group were incubated without any treatment, and each treatment was repeated twice.
Pouring the treated egg grains and the treatment liquid into a locust egg hatching device, and then placing the hatching device into an artificial climate box with RH of 50% and 39 ℃ and without illumination for culture. The hatchability of the daily locusts was recorded.
TABLE 11 statistical Table of the influence of octanoic acid on the hatchability of locusts in east Asia
Note that: transferring the hatched nymphs, and carrying out other experimental purposes.
From the above table, it is clear that the octanoic acid solutions with the concentrations of 10g/L and 1g/L have the effect of inhibiting hatching of the eggs of the migratory locust, and the hatching rate of the eggs of the migratory locust after the concentration treatment of 10g/L is lower.
2. Experiment for killing migratory locust in east Asia with different ages by octanoic acid
Preparing octanoic acid into octanoic acid solutions with different concentrations through a solvent system; the solvent system comprises absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system.
Experimental group:
1) 10mL of octanoic acid solution with the concentration of 10 g/L; 2) 10mL of octanoic acid solution with the concentration of 1 g/L;
control group: solvent control (absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system) and blank treatment are used as control groups.
Selecting 1-instar larvae, 3-instar nymphs and 5-instar adults of the migratory locust; spraying the treatment liquid to the locusts, uniformly wetting, transferring to a feeding device, each treatment is repeated for 5 times for 30 times, feeding the locusts by using tender corn leaves, grass leaves and clear water at room temperature (20-25 ℃), and placing cotton balls fully absorbed with water in the device to regulate the humidity and supplement water. The death of migratory locust was recorded for 24 h.
TABLE 12 statistics of killing Rate of octanoic acid on migratory locust in east Asia at each age (%)
As can be seen from the above table, the octanoic acid solutions at concentrations of 10g/L and 1g/L had extremely strong killing activity against migratory locust in any age, and the killing activity was positively correlated with the concentration.
EXAMPLE 4 incubation inhibition of Decoct egg of east Asia and killing experiments of different age locust by capric acid
1. Experiments of the hatching inhibition of decanoic acid on the eggs of migratory locust in east Asia
And (3) preparation of a medicament: preparing capric acid solutions with different concentrations by using a solvent system; the solvent system comprises absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system.
Experimental group:
1) 10mL of capric acid solution with the concentration of 10 g/L;
2) 10mL of capric acid solution with the concentration of 1 g/L;
Control group: the solvent control group (the solvent system is absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system) and the blank experiment are used as the control group.
Egg grain treatment: and taking fresh locusts in east Asia, peeling off the outer membrane of the locusts, only leaving locusts in east Asia, separating the locusts one by one, and ensuring the integrity of the locusts without mechanical damage to the locusts in the whole process.
And (3) medicament treatment: pouring the prepared treatment group solution and solvent contrast into culture dishes respectively, then placing fresh migratory locust egg grains, placing 300 egg grains into each culture dish, and soaking for 10min; the eggs of the blank group were incubated without any treatment, and each treatment was repeated twice.
Pouring the treated egg grains and the treatment liquid into a locust egg hatching device, and then placing the hatching device into an artificial climate box with RH of 50% and 39 ℃ and without illumination for culture. The hatchability of the daily locusts was recorded.
TABLE 13 statistical Table of the effect of decanoic acid on the hatchability of the eggs of migratory locust in east Asia (%)
Note that: transferring the hatched nymphs, and carrying out other experimental purposes.
From the above table, it is clear that the capric acid solutions with the concentrations of 10g/L and 1g/L have the effect of inhibiting hatching of the eggs of the migratory locust, and the hatching rate of the eggs of the migratory locust after the treatment with the concentration of 10g/L is the lowest.
2. Experiment for killing migratory locust in east Asia with different ages by capric acid
Preparing 1g/L and 10g/L capric acid solution of capric acid through a solvent system respectively; the solvent system comprises absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system.
Experimental group:
1) 10mL of capric acid solution with the concentration of 10 g/L; 2) 10mL of capric acid solution with the concentration of 1 g/L;
control group: solvent control (absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system) and blank treatment are used as control groups.
Selecting 1-instar larvae, 3-instar nymphs and 5-instar adults of the migratory locust; spraying the treatment liquid to the locusts, uniformly wetting, transferring to a feeding device, each treatment is repeated for 5 times for 30 times, feeding the locusts by using tender corn leaves, grass leaves and clear water at room temperature (20-25 ℃), and placing cotton balls fully absorbed with water in the device to regulate the humidity and supplement water. The death of migratory locust was recorded for 24 h.
TABLE 14 statistics of killing Rate of decanoic acid on east Asia migratory locust at each age (%)
As can be seen from the above table, the capric acid solutions at concentrations of 10g/L and 1g/L had extremely strong killing activity against migratory locust in any age, and the killing activity was positively correlated with the compound concentration.
Example 5 killing experiments of azelaic acid, cork acid, myristic acid and lauric acid on migratory locust of different ages
1. Active Compound preparation experiments
(1) Octanoic acid
Performing solid-liquid separation on the adult locusts in east Asia, removing suspended matters, and leaving supernatant; adding petroleum ether with the same volume into the supernatant to obtain a mixed solution 1; distilling the mixed solution 1 at about 40 ℃ under normal pressure, and recovering petroleum ether to leave a mixed solution 2; and (3) carrying out multistage reduced pressure rectification on the mixed solution 2, controlling the pressure in a rectifying tower to be 2KPa, and rectifying at the tower top temperature of 130-140 ℃ to obtain the octanoic acid.
(2) Capric acid
Pouring the adult locusts in the east Asia into a beaker, preheating to 70 ℃, slowly adding concentrated sulfuric acid under stirring, wherein the volume ratio of the extract to the concentrated sulfuric acid is 1:0.02, stopping stirring for 5min, preserving heat for 20min, pouring into a separating funnel, and washing with hot water at 70 ℃ until the mixture is neutral; preheating to 75 ℃, adding 1mol/L NaOH solution with the same volume as the extract under stirring, adding 5mol/L NaOH solution after stirring uniformly, and continuing stirring until the added NaOH solution enables the pH value of the system to be more than 12. Adding 35% dilute sulfuric acid into the system twice, uniformly stirring, measuring the pH of the system until the pH of the system is less than 2, continuously stirring until the pH is slightly boiling for 5min, stopping, pouring the upper layer into a separating funnel after the system is layered, washing with hot water at 75 ℃ to be neutral, and drying to obtain mixed fatty acid; the rectification is carried out by adopting a vacuum rectification device with a metal packing ring rectification column, and the process conditions are as follows: adding 600ml of mixed fatty acid obtained by hydrolysis into a 1000ml three-neck flask, continuously starting a rectifying tower, controlling the temperature at the bottom of the tower to 260 ℃, the temperature at the top of the tower to 120 ℃, the residual pressure to 600-1000Pa, and carrying out total reflux for 1h, and regulating the reflux ratio to 2:1, controlling the temperature of the tower top to 130 ℃, and collecting fractions, namely the capric acid.
(3) Cork acid
Extracting the adult locusta migratoria by using an equal volume of ethyl acetate as an extractant for 3 times, combining the extracts, and concentrating under reduced pressure until no fraction is obtained to obtain an ethyl acetate phase extract; dissolving ethyl acetate phase extract with ethanol, adsorbing on silica gel (200-300 meshes), subjecting to silica gel column chromatography (2.0 kg,10.0cm×80 cm), gradient eluting with chloroform-methanol system (100:0, 95:5, 90:10, 24L, v/v respectively), subjecting the eluate to TLC detection, mixing the same fractions to obtain Fr.1-Fr.5, and repeatedly recrystallizing Fr.1 with water and 50% ethanol to obtain cork acid.
(4) Azelaic acid
Extracting the adult locusta migratoria extract with equal volume of n-butanol as extractant for 3 times, mixing the extractive solutions, and concentrating under reduced pressure until no fraction is obtained to obtain n-butanol phase extract; the n-butanol phase extract was subjected to silica gel column chromatography, and eluted with a gradient of ethyl acetate-methanol (1:0, 20:1, 10:1,5:1,2:1,1:1,1:2, 0:1) to obtain 8 fractions (Fr 1-Fr 8), and the obtained fractions were repeatedly separated by silica gel column chromatography and recrystallized a plurality of times to obtain azelaic acid from FrC.
(5) Myristic acid
Extracting the adult locusts in east Asia with equal volume of ethyl acetate as extractant for 3 times, mixing the extractive solutions, and concentrating under reduced pressure to obtain ethyl acetate phase extract; subjecting the ethyl acetate phase extract to chromatography with reversed phase silica gel column, collecting, detecting, mixing, concentrating under reduced pressure to dry to obtain myristic acid primary crude product; subjecting the obtained primary myristic acid crude product to chromatographic treatment by a Sephadex LH-20 column, collecting, detecting, combining, concentrating under reduced pressure to dryness to obtain a secondary myristic acid crude product; subjecting the obtained secondary myristic acid crude product to chromatography with reversed phase silica gel column, collecting, detecting, mixing, concentrating under reduced pressure to dry to obtain primary myristic acid fine product; subjecting the obtained primary refined myristic acid to chromatography with normal phase silica gel column, collecting, detecting, mixing, concentrating under reduced pressure to dry to obtain secondary refined myristic acid; and (3) carrying out PTLC on the secondary refined myristic acid product, spotting, spreading, scraping, recovering a sample, and storing at-20 ℃ to obtain the myristic acid.
(6) Lauric acid
Pouring the adult locusts in the east Asia into a beaker, preheating to 70 ℃, slowly adding concentrated sulfuric acid under stirring, wherein the volume ratio of the extract to the concentrated sulfuric acid is 1:0.02, stopping stirring for 5min, preserving heat for 20min, pouring into a separating funnel, and washing with hot water at 70 ℃ until the mixture is neutral; preheating to 75 ℃, adding 1mol/L NaOH solution with the same volume as the extract under stirring, adding 5mol/L NaOH solution after stirring uniformly, and continuing stirring until the added NaOH solution enables the pH value of the system to be more than 12. Adding 35% dilute sulfuric acid into the system twice, uniformly stirring, measuring the pH of the system until the pH of the system is less than 2, continuously stirring until the pH is slightly boiling for 5min, stopping, pouring the upper layer into a separating funnel after the system is layered, washing with hot water at 75 ℃ to be neutral, and drying to obtain mixed fatty acid; the rectification is carried out by adopting a vacuum rectification device with a metal packing ring rectification column, and the process conditions are as follows: adding 600ml of mixed fatty acid obtained by hydrolysis into a 1000ml three-neck flask, continuously starting a rectifying tower, controlling the temperature of the top of the tower to be 150 ℃, collecting fractions until the temperature of the top of the tower continuously decreases, and stopping rectifying to obtain lauric acid.
2. Experiment of killing migratory locust in east Asia with different ages by active Compound
And (3) preparation of a medicament: preparing 10g/L solution of caprylic acid, capric acid and azelaic acid obtained from the market and obtained in the experiment through a solvent system; the solvent system is absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5 percent of the volume of the solvent system, the tween 80 accounts for 5 percent of the volume of the solvent system, and the water accounts for 90.5 percent of the volume of the solvent system;
preparing 10g/L cork acid, myristic acid and lauric acid obtained from the market and the experiment through a solvent system; the solvent system is absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 5.6 percent of the volume of the solvent system, the tween 80 accounts for 5 percent of the volume of the solvent system, and the water accounts for 89.4 percent of the volume of the solvent system.
Selecting 1-instar larvae, 3-instar nymphs and 5-instar adults of the migratory locust; spraying the treatment liquid to the locusts, uniformly wetting, transferring to a feeding device, each treatment is repeated for 5 times for 30 times, feeding the locusts by using tender corn leaves, grass leaves and clear water at room temperature (20-25 ℃), and placing cotton balls fully absorbed with water in the device to regulate the humidity and supplement water. The killing condition of the migratory locust in east Asia is recorded for 24 hours.
Table 15 shows statistics of killing rate (%) of active compounds prepared in Table 15 against migratory locust in east Asia at each age
Table 16 statistical table of the killing rate (%) of commercially available active compounds against migratory locust in east asia at each age
As can be seen from tables 15 to 16, the above-mentioned active compounds have the killing activity of octanoic acid, decanoic acid, lauric acid, myristic acid, cork acid, azelaic acid in the order from strong to weak; the table also shows that the commercial fatty acid has little difference with the active compound obtained by further treatment from the extract of the migratory locust in east Asia in killing effect on the migratory locust in east Asia, and the tolerance of the migratory locust in east Asia is enhanced and the killing activity of the compound is relatively weakened along with the increase of the age of the migratory locust in east Asia.
Example 6 experiment of Desmodium Ovata, caprylic acid, capric acid on the Dongya migratory locust
Respectively preparing 10g/L solution of caprylic acid and capric acid through a solvent system; the solvent system comprises absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system.
Selecting 3-year-old social locusts, namely the nymphs of the locusts as test insects, starving for 12 hours, measuring by adopting selective antifeedant activity, cutting 6g of corn leaves with consistent growth conditions, soaking the corn leaves in the treatment liquid for 30 seconds (or uniformly spraying the leaves without dropping the treatment liquid), weighing the leaves with the same quality for soaking in distilled water in contrast, taking out, and naturally airing. The prepared leaves are respectively placed at the center positions of two insect-raising cages with the length of 30 x 30cm, 30 locusts in east Asia with the age of 3 are placed in each cage, the two cages are connected in a seamless mode, the locusts can pass through freely, the leaves are taken out after 3 hours, the feeding weight of the corn leaves is measured (blank control is arranged at the same time to remove the weight loss of the corn leaves), and each treatment is repeated for 5 times. Meanwhile, the distribution condition of the migratory locust in two cages is observed.
Table 17 statistical table of data relating to the avoidance of migratory grasshopper
From the above table, it was found that there were 29 migratory grasses in the cage of the corn leaf treated with octanoic acid, 31 migratory grasses in the cage of the corn leaf treated with clear water, and the decanoic acid results were not different from the above average, and the calculated evasion rates of octanoic acid and decanoic acid were about 3% and 6%, and the evasion effects of the octanoic acid and decanoic acid on the migratory grasses were not obvious.
Example 7 test of killing 3-year-old nymphs of migratory locust with Compound pesticide
And (3) preparation of a medicament:
(1) Treatment 1: dissolving 2g of permethrin (active ingredient 25%) in 30ml of distilled water to obtain a solution I, weighing 0.5g of caprylic acid or capric acid or a mixture of caprylic and capric acid (mass ratio of 1:1), dissolving in 2.25ml of absolute ethyl alcohol, adding 0.2g of BHT into the solution, adding 2.5ml of Tween 80 into the solution after dissolving, adding 15.25ml of distilled water to obtain a solution II, mixing and shaking the solution I and the solution II uniformly, wherein the final concentration of permethrin is 10g/L, and the final concentration of fatty acid is 10g/L;
(2) Treatment 2: weigh 8.333×10 -3 And g chlorfenapyr (active ingredient 24%) is dissolved in 49.85ml distilled water to obtain a solution I, 0.04g of caprylic acid or capric acid or a mixture of caprylic and capric acid (mass ratio of 1:1) is additionally weighed and dissolved in 2ml dimethyl sulfoxide to prepare a solution II with the concentration of 20mg/ml, 0.15ml is added into the solution I and uniformly mixed, wherein the final concentration of chlorfenapyr is 0.04mg/ml, and the final concentration of fatty acid is 0.06mg/ml.
(3) Treatment 3: 0.5g of octanoic acid is weighed and dissolved in 2.25ml of absolute ethyl alcohol, 2.5ml of Tween 80 is added into the solution, 45.25ml of distilled water is added into the solution after shaking the solution evenly, and the final concentration of fatty acid is 10g/L.
(4) Treatment 4: 0.5g of decanoic acid is weighed and dissolved in 2.25ml of absolute ethyl alcohol, 2.5ml of tween 80 is added into the absolute ethyl alcohol, 45.25ml of distilled water is added into the absolute ethyl alcohol after shaking evenly, and the absolute ethyl alcohol is evenly mixed, so that the final concentration of fatty acid is 10g/L.
Control group: solvent control (absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system) and blank treatment are used as control groups.
Selecting a plurality of 3-instar nymphs of the migratory locust; spraying the treatment liquid to the locusts, uniformly wetting, transferring to a feeding device, each treatment is repeated for 5 times for 30 times, feeding the locusts by using tender corn leaves, grass leaves and clear water at room temperature (20-25 ℃), and placing cotton balls fully absorbed with water in the device to regulate the humidity and supplement water. Recording the subsequent killing condition of the migratory locust in east Asia for 24 hours.
Table 18 statistics of killing rate of compound pesticide on migratory locust in east Asia (%)
| Numbering device | Treatment group | Locust mortality (%) | Correction of mortality (%) |
| 1 | Treatment 1: permethrin + fatty acid | 100 | 100 |
| 2 | Treatment 2: chlorfenapyr + fatty acid | 40 | 33 |
| 3 | Treatment 3: octanoic acid | 100 | 100 |
| 4 | Treatment 4: capric acid | 100 | 100 |
| 5 | Solvent control | 7 | -3 |
| 6 | Blank control | 10 | 0 |
The comparison of the biological activities of the compound pesticide and the fatty acid on the migratory locust in east Asia is shown in the table above, wherein the mortality rate of the permethrin, the fatty acid composition and the fatty acid used alone reaches 100% after 30min of treatment, and the chlorfenapyr+fatty acid composition does not show stronger biological activity; in conclusion, the fatty acid does not only have an auxiliary agent function, and the fatty acid alone can generate extremely strong killing activity on the migratory locust in east Asia.
Examples 8 and Huang Jizhu experiment of killing of locust extracts and octanoic acid on different ages
1. Experiment of killing different-age locust with Huang Jizhu locust extract
(1) Preparation of Huang Jizhu locust extract
Extracting crushed Huang Jizhu adult locust grease (volume ratio petroleum ether: huang Jizhu adult locust=1:3) with petroleum ether, extracting at 75deg.C for 2h, reflux-extracting defatted Huang Jizhu adult locust with 85% ethanol (volume ratio defatted Huang Jizhu adult locust: 85% ethanol=1:10), and reflux-extracting for 2h after residual petroleum ether is volatilized; obtaining an extracting solution, and concentrating the extracting solution by rotary evaporation to obtain a small-volume concentrated solution, namely Huang Jizhu adult locust extract; sequentially extracting the concentrated solution with petroleum ether, ethyl acetate and n-butanol with equal volume, and concentrating the petroleum ether phase, ethyl acetate phase and n-butanol phase by rotary evaporation respectively to obtain petroleum ether liquid extract, ethyl acetate liquid extract and n-butanol liquid extract.
(2) Experiment of killing different-age locust with Huang Jizhu locust extract
The petroleum ether extract, ethyl acetate extract and n-butanol extract of the adult locusts of Huang Jizhu are respectively prepared into 10% solutions by water, and meanwhile, a clear water treatment group and a blank control group are taken for a control experiment. Huang Jizhu locust 1-age larvae, 3-age nymphs and 5-age adults hatched on the same day in a blank control group are selected, the nymphs are sprayed by adopting solutions of each treatment group, every 30 larvae are put into a feeding device, every group is repeated 5 times, fresh corn leaves and clear water are replaced every day for feeding, the locust growth conditions of each group are counted, and the counting results after 7 days are shown in the following table.
Table 19 Huang Jizhu locusts extract statistics of killing rate of Huang Jizhu locusts (%)
From the above table, it can be seen that the petroleum ether extract and ethyl acetate extract of Huang Jizhu locusts at concentrations of 5% and 10% also have better killing activity against Huang Jizhu locusts at ages 1, 3 and adults.
2. Experiment for killing locust of different ages with caprylic acid and capric acid
And (3) preparation of a medicament: respectively preparing 10g/L solution of caprylic acid and capric acid through a solvent system; the solvent system comprises absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system.
Experimental group:
1) 10mL of octanoic acid solution with the concentration of 10 g/L;
2) 10mL of capric acid solution with the concentration of 10 g/L;
control group: solvent control (absolute ethyl alcohol, tween 80 and water, wherein the absolute ethyl alcohol accounts for 4.5% of the volume of the solvent system, the tween 80 accounts for 5% of the volume of the solvent system, and the water accounts for 90.5% of the volume of the solvent system) and blank treatment are used as control groups.
Selecting Huang Jizhu locust 1-age larvae, 3-age nymphs and 5-age adults; spraying the treatment liquid to the locusts, uniformly wetting, transferring to a feeding device, each treatment is repeated for 5 times for 30 times, feeding the locusts by using tender corn leaves, grass leaves and clear water at room temperature (20-25 ℃), and placing cotton balls fully absorbed with water in the device to regulate the humidity and supplement water. The killing of Huang Jizhu locusts was recorded for 24 hours.
TABLE 20 statistics of killing Rate of caprylic acid and capric acid solutions on different ages of Huang Jizhu locusts (%)
From the table, the caprylic acid and capric acid solutions with the concentration of 10g/L have extremely strong killing activity on Huang Jizhu locusts in any age.
Claims (9)
1. An application of locust extract as anti-locust agent, which is characterized in that:
the locust extract is obtained by degreasing adult locust or ovum thereof as raw material, extracting with organic solvent after degreasing, concentrating the extractive solution, and sequentially extracting with organic solvent;
The degreasing organic solvent is petroleum ether; the volume ratio of the organic solvent to the adult locust or the eggs thereof is 1:3;
the organic solvent is 85% ethanol, and the volume ratio of the defatted locust adults or the worm eggs to the 85% ethanol is 1:10;
the organic solvent adopted in the extraction is 1% -10% petroleum ether, 1% -10% ethyl acetate and 1% -10% n-butanol.
2. The use according to claim 1, characterized in that: the active component of the locust extract is one or more of myristic acid, lauric acid, capric acid and n-caprylic acid.
3. An anti-locust agent, characterized in that: the active ingredient of the locust resistant agent is the locust extract as claimed in claim 1.
4. A locust resistant agent according to claim 3, characterised in that: the locust resistant agent is prepared by mixing an active ingredient with an agriculturally acceptable carrier; wherein the active ingredient accounts for 1-10% of the weight of the locust resistant agent.
5. An anti-locust composition, characterized in that: the composition comprises a component A and a component B, wherein the component A and the component B are mixed according to the weight part ratio of 1:100-100:1; the component A is the locust extract of claim 1; component B is other compounds and/or pesticides with insecticidal, bactericidal, virucidal, chemically sterile, mating disruption properties.
6. An anti-locust composition formulation, characterized in that: mixing the locust resistant composition according to claim 5 as active ingredient with an agriculturally acceptable carrier; wherein the active ingredient accounts for 0.01-99% of the weight of the locust-resistant composition.
7. A use of a formulation characterized by: use of an anti-locust agent according to claim 3 or an anti-locust composition formulation according to claim 6 for protecting plants against locust stress.
8. Use of a formulation according to claim 7, wherein: the use of said formulation for protecting plants against the stress of migratory grasses in east asia.
9. A method of combating damage caused by a locust, comprising: the locust extract of claim 1; or, an anti-locust agent according to claim 3; or, the locust-resistant composition preparation according to claim 6 is applied to any area through which the locust passes, thereby acting as a locust-resistant agent.
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Citations (2)
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| CN101861095A (en) * | 2007-09-14 | 2010-10-13 | Smg品牌公司 | Pest control compositions, and methods and products utilizing same |
| CN104093307A (en) * | 2012-03-06 | 2014-10-08 | 花王株式会社 | Method for controlling pests |
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| US8142801B2 (en) * | 2009-02-02 | 2012-03-27 | Ecoblend, Llc | Pesticidal compositions and methods of use thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101861095A (en) * | 2007-09-14 | 2010-10-13 | Smg品牌公司 | Pest control compositions, and methods and products utilizing same |
| CN104093307A (en) * | 2012-03-06 | 2014-10-08 | 花王株式会社 | Method for controlling pests |
Non-Patent Citations (1)
| Title |
|---|
| 飞蝗及沙漠蝗信息素概述;李嘉;张龙;;应用昆虫学报(第04期);第1046-1051页 * |
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