CN104738034A - Acetic acid cis-3-hexenyl ester synergistic holotrichia parallela sex attractant - Google Patents
Acetic acid cis-3-hexenyl ester synergistic holotrichia parallela sex attractant Download PDFInfo
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Abstract
The invention discloses an acetic acid cis-3-hexenyl ester synergistic holotrichia parallela sex attractant, wherein the weight ratio of the sex attractant and acetic acid cis-3-hexenyl ester is (3-7):1. The sex attractant comprises a holotrichia parallela sex pheromone component and an organic solvent, wherein the holotrichia parallela sex pheromone component comprises methyl L-isoleucinate, (R)-(-)-linalool and 1-aminocyclohexanecarboxylic acid in a weight ratio of 6: 1: 0.1 and the weight ratio of the holotrichia parallela sex pheromone component and the organic solvent is 1: 3. The organic solvent is one or more of dichloromethane, methanol or acetone. Behavior experiments show that EAG and selective reaction rate of male and female worms on a mixture of the sex attractant and acetic acid cis-3-hexenyl ester are relatively good. The field trapping experiments on holotrichia parallela show that acetic acid cis-3-hexenyl ester has a synergistic effect on the trapping effect of the sex attractant, thereby providing a ground for monitoring and preventing and treating holotrichia parallela by using plant volatile matters and insect sex pheromone in a synergistic manner.
Description
Technical field
The present invention relates to a kind of insect sex attractant of botanical components synergy, specifically, relate to the Holotrichia parallela gyplure of a kind of cis-3-hexene ester synergy.
Background technology
Holotrichia parallela (Holotrichia parallela) belongs to coleoptera (Coleoptera) scarab beetle Superfamily (Scarabaeoidea) Melolonthidae (Melolonthidae), is that during agriculture forest and husbandry is produced, the important soil of a class is dwelt insect.The various crop such as the harm of its larva grub peanut, soybean, pachyrhizus, corn, lawn, flowers, vegetables, often cause huge economic loss (Hu Qiongbo, 2004; Liu Shusen etc., 2008).For peanut, peanut grub generation quantity in all parts of the country is in recent years in ascendant trend year by year, and cause serious harm to peanut production, be generally injured underproduction 10-20%, severe patient 50-80%, and even No kernels or seeds are gathered, as in a year of scarcity (Li Xiao etc., 2012).Because Holotrichia parallela is for a long time at underground activities, therefore carrying out prevention and control to it has certain difficulty, and present stage is still based on chemical control.In addition, residue of pesticide, the problems such as environmental pollution exist, and make us carry out pest control in the urgent need to a kind of novel approach of green non-pollution.
The use of insect sex pheromone in recent years provides a new approach for control of insect.Adopt insect sex pheromone to carry out worm monitoring and non-environmental pollution control and there are active high, high specificity, nontoxic, pollution-free, not killed natural enemies, advantage (Nehme et al., 2009 such as easy to use, with low cost; Nehme et al., 2010; Wang Guihua etc., 2013).The sex pheromone of Shandong Peanut Inst. to China's Holotrichia parallela is studied, independent research holotrichia parallela sex pheromone core and supporting trapper, and achieves in field and good lure worm effect (Brockerhoff et al., 2006; Zhou et al., 2009; Li Xiao etc., 2012).But there are problem (Reddy and Guerrero, 2004 such as specific attractant male worm, consumption time be short in gyplure;
et al., 2005).In recent years, along with people deepen day by day to understanding chemically interactive between insect and plant, utilize Chemical lab management principle, the functional compounds such as utilization Secondary metabolites regulate, control pest obtains significant progress (Dickens et al., 1993 with the research of containment objective plant; Dodds, 2011; Yang Hui etc., 2008; Yang Lang etc., 2004).Plant volatiles and insect pheromone are the important information materials of insect, and the two synergy is with the behavior (Deng et al., the 2004a that regulate insect; Hanks and Millar, 2012a; Jung et al., 2013).By increasing tentaculum electric potential, pheromones receives neuron action potential and pulse frequency, and specialized plant source volatile matter significantly can strengthen allure (Reddy and Guerrero, 2004 of insect sex pheromone; Wang Zhenhua etc., 2008).Research shows, host plant self produces and the secondary pollutant mass-energy discharged regulation and control plant-feed insect multiple behavior, such as orientation, mate and oviposit, take food, escape, assemble, behavior (Fettig et al., 2012, Lait et al., 2003 such as pollination; Pan et al., 2013; Wu Limin etc., 2012).But in earlier stage about the research of insect attractant mainly concentrates on plant volatiles and the one-sided application of insect pheromone in control of insect (Negri and Bernik, 2008; Szendrei et al., 2011, Tooker et al., 2002).Many bibliographical informations in recent years, botanical components and insect pheromone are worked in coordination with and are played regulating and controlling effect to insect lipids, prove that plant volatiles has remarkable synergistic effect (Douglas et al .1993 to examination worm gyplure; Lu Yujie etc., 2003; Wang Guihua etc., 2013).
By the new technology that sex pheromone or gyplure pest control are a kind of non-environmental pollution control insects.Although existing hundreds of insect sex attractant is applied both at home and abroad, Qu Mingjing is applied for a kind of Holotrichia parallela sex pheromone and gyplure in the Chinese patent of 200910162405.4 at application number, on this basis, how by botanical components and insect pheromone efficient combination, realizing botanical components lures worm effect synergistic effect to examination worm gyplure, is the technical problem to be solved in the present invention.
Summary of the invention
In order to solve problems of the prior art, the object of the present invention is to provide a kind of Holotrichia parallela gyplure utilizing the synergy of cis-3-hexene ester, the potentiometric response that cis-3-hexene ester can cause female male worm stronger and olfaction reaction; There is synergistic function to gyplure in Holotrichia parallela field trapping test display cis-3-hexene ester, monitors and prevent and treat Holotrichia parallela provide foundation for utilizing plant volatiles and insect sex pheromone.
The technical scheme that the present invention takes is:
The Holotrichia parallela gyplure of a kind of cis-3-hexene ester synergistic effect, it is characterized in that, the weight ratio of gyplure and cis-3-hexene ester is (3-7): 1, described gyplure comprises Holotrichia parallela sex pheromone component and organic solvent, described Holotrichia parallela sex pheromone component comprises ILE methyl esters, (R)-(-)-linalool and 1-cyclohexane-carboxylic acid, its weight proportion is 6:1:0.1, and described Holotrichia parallela sex pheromone component and organic solvent weight ratio are 1:3, described organic solvent is carrene, one or more in methyl alcohol or acetone.
The Electroantennogram response test method of the Holotrichia parallela gyplure of cis-3-hexene ester synergistic effect, is characterized in that, specifically comprise the steps:
(1) preparation of stimulated samples
The preparation of plant volatiles single dose: each compound all uses methanol as solvent, is configured to 0.1,1,10 μ g/ μ l, 3 concentration, compare with methyl alcohol;
The preparation of plant volatiles and gyplure mixture: pheromones component ILE methyl esters, (R)-(-)-linalool and 1-cyclohexane-carboxylic acid are that 6:1:0.1 ratio is hybridly prepared into Holotrichia parallela gyplure according to weight proportion by this patent, with the mother liquor of 1 μ g/ μ l, according to gyplure: the ratio mixing of plant volatiles=1:1,3:1,5:1,7:1,9:1, compounding dose of institute is for indoor electric physiology and performance testing;
(2) feeler process and EAG test program
The each chemical reagent sample of 10ul is extracted with microsyringe, be added drop-wise to (25mm × 8mm) on doubling filter paper, with tweezers, filter paper is put into Pasteur pipe after candidate agent volatilization 5min, ParafilmTM Pasteur pipe two ends, sample volatilizees after 5min and measures in pipe, it is 500ms that the phase is gone through in stimulation, stimulus intervals is about 30s, the flow velocity of continuous flow is all 4mL/s with stimulating the flow velocity of air-flow, every cephalont only takes 1 feeler, and 6 feelers are tested in each process, and every root feeler repeats 3 times, contrast hockets with the mensuration of processed group, take methyl alcohol as blank.
Further, described gyplure: the mixture formed when cis-3-hexene ester is 3:1, Electroantennogram signal (EAG) response value of female worm is 2.35 ± 0.56mV.
Further, described gyplure: the mixture formed when cis-3-hexene ester is 5:1, Electroantennogram signal (EAG) response value of male worm is 3.32 ± 0.55mV.
The olfaction reaction method of testing of the Holotrichia parallela gyplure of cis-3-hexene ester synergistic effect, is characterized in that, specifically comprise the steps:
(1) preparation of stimulated samples
The preparation of plant volatiles single dose: each compound all uses methanol as solvent, is configured to 0.1,1,10 μ g/ μ l, 3 concentration, compare with methyl alcohol;
The preparation of plant volatiles and gyplure mixture: pheromones component ILE methyl esters, (R)-(-)-linalool and 1-cyclohexane-carboxylic acid are that 6:1:0.1 ratio is hybridly prepared into Holotrichia parallela gyplure according to weight proportion by this patent, with the mother liquor of 1 μ g/ μ l, according to gyplure: the ratio mixing of plant volatiles=1:1,3:1,5:1,7:1,9:1, compounding dose of institute is for indoor electric physiology and performance testing;
(2) Holotrichia parallela is measured to the susceptibility size of various reagent by " Y " type olfactometer, olfactometer is placed in darkroom, room temperature 27 DEG C, air velocity 500mL/min, get each reagent dropwise of 10ul in test on filter paper bar, and put into " Y " type pipe side arm air inlet place, methyl alcohol contrasts, often group exchanges experimental group and control group seat after repeating, and avoids experimental error; The filter paper bar renewed after each test, often kind of reagent repeats for 3 times, and rear replacing " Y " type pipe surveyed by reagent of the same race, with the effective acetone cleaning of " Y " type crossed, 60 DEG C of oven dry; Often group test picking 10 was unearthed and active strong examination worm the same day, put into " Y " type pipe principal arm port, observed examination worm distribution situation after 10min, and calculate reactivity, relative response rate, reaction coefficient carry out data analysis;
Reactivity=[(in taste source capsule in borer population+control tube borer population)/test total borer population];
Select reactivity=[borer population in taste source capsule/(in taste source capsule in borer population+control tube borer population)].
Further, described gyplure: the mixture formed when cis-3-hexene ester is 3:1, the selection reactivity of female worm is the highest, is 0.90 ± 0.01.
Further, described gyplure: the mixture formed when cis-3-hexene ester is 3:1, the selection reactivity of male worm is the highest, is 0.93 ± 0.07.
The field of the Holotrichia parallela gyplure of cis-3-hexene ester synergy lures worm test method, it is characterized in that, specifically comprise the steps: to select field trial base, every 60m arranges a trapper, described trapper is by top cover, funnel type device, intersection baffle plate and gathering unit composition, the baffle plate that wherein intersects is located at below top cover, funnel device is located at and is intersected between baffle plate, gathering unit is located at below funnel device, the suspension height of trapper (top) is 2.0m, with the mother liquor of 360ug/ul, gyplure and cis-3-hexene ester are mixed with 1:1, 3:1, 5:1, 7:1, 9:1 five different proportions lure core, the described core carrier main component that lures is absorbent cotton, concentration is that 360mg/ lures core, core is lured to concentrate freezen protective at refrigerator, respectively with cis-3-hexene ester and gyplure as a control group, often lure the repetition of three, core every day, choose five unearthed day data analysis.After trapping every day terminates, record each trapper and lure borer population amount, calculate and lure core to the synergistic effect of gyplure, computational methods are as follows:
Further, described gyplure: during cis-3-hexene ester=5:1, often lure core to lure female borer population to reach maximum every day to be 11.00 ± 0.76, to be significantly higher than 2.67 ± 0.33 of gyplure, the synergistic effect of cis-3-hexene ester to gyplure reaches 311%.
Further, described gyplure: during cis-3-hexene ester=3:1, often lure core to lure male worm number to reach maximum every day to be 285.00 ± 8.44, to be significantly higher than 116 ± 2.65 of gyplure, the synergistic effect of cis-3-hexene ester to gyplure reaches 146%.
Beneficial effect of the present invention is:
Attempt Holotrichia parallela gyplure to combine with its host plant volatile matter herein, thus reach and reduce costs, improve the objects such as pest trap rate.The host plant of Holotrichia parallela mainly contains elm, mulberry tree etc., by the active ingredient of contrast host plant leaf volatiles matter, pick the multiple frequently seen plants source volatile matter from Holotrichia parallela host plant blade herein, by Electroantennogram response (electroantennogram, EAG), and combine " Y " type olfactometer and filter out Holotrichia parallela than more sensitive chemical substance, the synergistic effect of Holotrichia parallela lured by test host plant volatile confrontation gyplure, result shows the potentiometric response that cis-3-hexene ester can cause female male worm stronger and olfaction reaction, after it being mixed in varing proportions with gyplure, it is better with selection reactivity to the EAG of mixture that Behavior Test shows female male worm.There is synergistic function in Holotrichia parallela field the lure collection of trapping test display cis-3-hexene ester to gyplure, when gyplure: during cis-3-hexene ester=5:1, often lure core to lure female borer population to reach maximum (11.00 ± 0.76) every day, 311% is reached to the synergistic effect of gyplure; For male worm, when gyplure: during cis-3-hexene ester=3:1, often lure and lure worm amount maximum (285.00 ± 8.44) core day, 146% is reached to the synergistic effect of gyplure.Result herein utilizes plant volatiles and insect sex pheromone to monitor and prevent and treat Holotrichia parallela to provide foundation for collaborative.
The Holotrichia parallela gyplure utilizing the synergy of cis-3-hexene ester of the present invention's research and development; no matter be used to do detecting and reporting pest information or directly prevent and treat Holotrichia parallela insect pest by mass trapping and Insect pest suppression method; all be conducive to the power giving full play to biological control; control efficiency is improved while Protect natural enemies; avoiding using chemical pesticide in addition can decreasing pollution; protection of the environment, economic benefit, social benefit and ecological benefits are remarkable.
Embodiment
1 materials and methods
1.1 for examination insect and reagent
For examination insect: Holotrichia parallela picks up from Laixi, Qingdao of Shandong province Wangcheng Shandong Peanut Inst. proving ground, after the Holotrichia parallela adult male and female of adopting back are separated, raise with fresh leaf of elm tree in the incubator having ventilation hole respectively, in incubator, soil moisture remains between 18%-20%.For examination insect Nature enemy 24 hours before the test.
For having a try agent: test agents useful for same is purchased from Sigma company, and each reagent purity is all more than 95%.Concrete reagent is in table 1.
Behavior reaction tester: (1) tentaculum electric potential instrument is made up of fine motion executor INR-5, data acquisition controller IDAC-4, Odor stimulation control device cs-05 and tentaculum electric potential record display translation device, originates from German Syntech company.(2) olfactometer consists of the following components: ACO-006 type electromagnetic type air compressor (production of Sheng Group Co., Ltd of Guangdong Province), activated carbon air purifying device and LZB-4 type glass rotameter (Changzhou Shuanghuan Thermo-Technical Instrument Co., Ltd.'s production), 250ml Te Shi porous filter flask (in-built water), three-way pipe, Y-shaped smell sensing apparatus (two brachium 12cm, the long 20cm of base tube, two-arm angle 45 °, caliber 3.5cm).
Lure core and trapper: lure core and the supporting trapper thereof that adopt in test provide by Shandong peanut Research Institute.Lure core carrier main component to be absorbent cotton, concentration is that 360mg/ lures core.Trapper is made up of top cover, funnel type device, intersection baffle plate and gathering unit, the baffle plate that wherein intersects is located at below top cover, funnel device is located at and is intersected between baffle plate, and gathering unit is located at below funnel device, has volatile core that lures by force and concentrates freezen protective at refrigerator.
Table 1 many kinds of host plant volatile matter
1.2 test method
1.2.1 the preparation of stimulated samples
The preparation of plant volatiles single dose: each compound all uses methanol as solvent, is configured to 0.1,1,10 μ g/ μ l, 3 concentration, compare with methyl alcohol.
The preparation of plant volatiles and gyplure mixture: pheromones component ILE methyl esters, (R)-(-)-linalool and 1-cyclohexane-carboxylic acid are that 6:1:0.1 ratio is hybridly prepared into Holotrichia parallela gyplure according to weight proportion by this patent, with the mother liquor of 1 μ g/ μ l, according to gyplure: the ratio mixing of plant volatiles=1:1,3:1,5:1,7:1,9:1, compounding dose of institute is for indoor electric physiology and performance testing.
1.2.2 feeler process and EAG test program
With reference to Holotrichia parallela feeler processing methods (Zhou et al., 2009) such as zhou.The each chemical reagent sample of 10ul is extracted with microsyringe, be added drop-wise to (25mm × 8mm) on doubling filter paper, with tweezers, filter paper is put into Pasteur pipe after candidate agent volatilization 5min, ParafilmTM Pasteur pipe two ends, sample volatilizees after 5min and measures in pipe, it is 500ms that the phase is gone through in stimulation, and stimulus intervals is about 30s.The flow velocity of continuous flow is all 4mL/s with stimulating the flow velocity of air-flow, and every cephalont only takes 1 feeler, and 6 feelers are tested in each process, and every root feeler repeats 3 times, and contrast hockets with the mensuration of processed group, take methyl alcohol as blank.
1.2.3 olfaction reaction test
This patent measures Holotrichia parallela to the susceptibility size of various reagent by " Y " type olfactometer.Olfactometer is placed in darkroom, room temperature 27 DEG C, air velocity 500mL/min.Get each reagent dropwise of 10ul in test on filter paper bar, and put into " Y " type pipe side arm air inlet place, methyl alcohol contrasts, and often group exchanges experimental group and control group seat after repeating, and avoids experimental error.The filter paper bar renewed after each test, often kind of reagent repeats for 3 times, and rear replacing " Y " type pipe surveyed by reagent of the same race, with the effective acetone cleaning of " Y " type crossed, 60 DEG C of oven dry.Often group test picking 10 was unearthed and active strong examination worm the same day, put into " Y " type pipe principal arm port, observed examination worm distribution situation after 10min, and calculate reactivity, relative response rate, reaction coefficient carry out data analysis.
Reactivity=[(in taste source capsule in borer population+control tube borer population)/test total borer population];
Select reactivity=[borer population in taste source capsule/(in taste source capsule in borer population+control tube borer population)].
1.2.4 the field of plant volatiles and gyplure mixture lures worm to test
Field trial is carried out in proving ground, Laixi, Qingdao.Every 60m arranges a trapper, and the suspension height of trapper (top) is 2.0m.With the mother liquor (Li Xiao etc. of 360ug/ul, 2012) what gyplure and cis-3-hexene ester are mixed with 1:1,3:1,5:1,7:1,9:1 five different proportions lures core, with plant volatiles matter single dose and gyplure as a control group, often lure the repetition of three, core every day, choose five unearthed day data analysis.After trapping every day terminates, record each trapper and lure borer population amount, calculate and lure core to the synergistic effect of gyplure.Computational methods are as follows:
1.5 data processing
Use spss 16.0 software to carry out computational analysis to data, Multiple range test adopts DuncanShi duncan's new multiple range method.
2. result
2.1 Holotrichia parallelas are to the electro physiology of various plants source volatile matter and behavior reaction
2.1.1 plant volatiles impact that Holotrichia parallela EAG is reacted
The female male worm of Holotrichia parallela is to EAG response value difference (table 2) of reagent variable concentrations of the same race.Female male worm all shows to cis-3-hexene ester the EAG response value being significantly higher than other reagent, and the EAG value of female worm to cis-3-hexene ester is 4.84 ± 0.37mV, and male worm is 4.02 ± 0.36mV.
Table 2 Holotrichia parallela reacts the EAG of host plant volatile matter
In table, data are mean ± standard error.Remarkable at P<0.05 level difference through the inspection of DuncanShi duncan's new multiple range method with letter representation different after column data.
* in table, numerical value is the EAG value using methyl alcohol as CK.
2.1.2 plant volatiles is on the impact of Holotrichia parallela olfaction reaction
As can be seen from Table 2, the reactivity of selected examination worm all more than 80%, and does not have significant difference between each process, illustrates that examination worm is healthy, this research experiment reliable results.As can be seen from selection reactivity, Holotrichia parallela is different to the response value of reagent variable concentrations of the same race.The selection reactivity of female worm to cis-3-hexene ester is the highest, is 0.98 ± 0.02, is significantly higher than other reagent of variable concentrations.For male worm, the otherness between each reagent is comparatively remarkable, and wherein the selection reactivity of male worm to cis-3-hexene ester is significantly higher than other reagent, is 0.73 ± 0.04.
Table 3 Holotrichia parallela is to the olfaction reaction * of host plant volatile matter
In table, data are mean ± standard error.Remarkable at P<0.05 level difference through the inspection of DuncanShi duncan's new multiple range method with letter representation different after column data.
* in table, numerical value is the selection reactivity using methyl alcohol as CK.
The synergism test of 2.2 host plant volatile confrontation gyplures
From 2.1, male and female Holotrichia parallela is to the EAG response value of cis-3-hexene ester and select reactivity all higher than other reagent, and the reagent of male and female Holotrichia parallela to 1ug/ul concentration has higher susceptibility, therefore this patent chooses cis-3-hexene ester, be mixed with the mixture (hereinafter referred to as mixture) of 1ug/ul respectively with gyplure by different proportion (m:m), carry out synergy test.
2.2.1 mixture is on the impact of Holotrichia parallela Electroantennogram response
The female male worm of Holotrichia parallela is to EAG response value difference (table 4) of different ratio group.When gyplure: when cis-3-hexene ester is 3:1, female worm EAG response value is significantly higher than other proportioning group, is 2.35 ± 0.56mV.Male worm is to the gyplure of 5:1: the EAG response value of cis-3-hexene ester mixture is significantly higher than other proportioning groups, is 3.32 ± 0.55mV.
The EAG of table 4 Holotrichia parallela to gyplure and host plant volatile matter mix reagent reacts
In table, data are mean ± standard error.Remarkable at P<0.05 level difference through the inspection of DuncanShi duncan's new multiple range method with letter representation different after column data.
* in table, numerical value is the EAG value using gyplure as CK.
2.2.2 mixture is on the impact of Holotrichia parallela olfaction reaction
As can be seen from Table 5, the reactivity of selected examination worm all more than 80%, and does not have significant difference between each process, illustrates that examination worm is healthy, this research experiment reliable results.As can be seen from selection reactivity, Holotrichia parallela is different to the selection reactivity of different ratio group.When gyplure: when cis-3-hexene ester is 3:1, the selection reactivity of female worm is the highest, is respectively 0.90 ± 0.01, is significantly higher than other proportioning groups.When gyplure: when cis-3-hexene ester is 3:1, the selection reactivity of male worm is significantly higher than other proportioning groups, is 0.93 ± 0.07.
Table 5 Holotrichia parallela is to the olfaction reaction * of gyplure and host plant volatile matter mix reagent
In table, data are mean ± standard error.Remarkable at P<0.05 level difference through the inspection of DuncanShi duncan's new multiple range method with letter representation different after column data.
* in table, numerical value is the selection reactivity using gyplure as CK.
2.3 field adult trap effects are analyzed
As shown in Table 6, the quantity of each mixture trapping Holotrichia parallela adult is all significantly higher than cis-3-hexene ester single dose or gyplure.When gyplure: during cis-3-hexene ester=5:1, often lure core to lure female borer population to reach maximum every day, be 11.00 ± 0.76, be significantly higher than 2.67 ± 0.33 of gyplure, the synergistic effect of cis-3-hexene ester to gyplure reaches 311%.For male worm, when gyplure: during cis-3-hexene ester=3:1, often luring and lure worm amount maximum core day, is 285.00 ± 8.44, is significantly higher than 116 ± 2.65 of gyplure, and the synergistic effect of cis-3-hexene ester to gyplure reaches 146%.
The field trap effect of table 6 Holotrichia parallela gyplure and plant volatiles mixture is analyzed
In table, data are mean ± standard error.Remarkable at P<0.05 level difference through the inspection of DuncanShi duncan's new multiple range method with letter representation different after column data.
Claims (10)
1. the Holotrichia parallela gyplure of cis-3-hexene ester synergistic effect, it is characterized in that, the weight ratio of gyplure and cis-3-hexene ester is (3-7): 1, described gyplure comprises Holotrichia parallela sex pheromone component and organic solvent, described Holotrichia parallela sex pheromone component comprises ILE methyl esters, (R)-(-)-linalool and 1-cyclohexane-carboxylic acid, its weight proportion is 6:1:0.1, and described Holotrichia parallela sex pheromone component and organic solvent weight ratio are 1:3, described organic solvent is carrene, one or more in methyl alcohol or acetone.
2. the Electroantennogram response Electroantennogram response test method of the Holotrichia parallela gyplure of cis-3-hexene ester synergistic effect according to claim 1, is characterized in that, specifically comprise the steps:
(1) preparation of stimulated samples
The preparation of plant volatiles single dose: each compound all uses methanol as solvent, is configured to 0.1,1,10 μ g/ μ l, 3 concentration, compare with methyl alcohol;
The preparation of plant volatiles and gyplure mixture: pheromones component ILE methyl esters, (R)-(-)-linalool and 1-cyclohexane-carboxylic acid are that 6:1:0.1 ratio is hybridly prepared into Holotrichia parallela gyplure according to weight proportion by this patent, with the mother liquor of 1 μ g/ μ l, according to gyplure: the ratio mixing of plant volatiles=1:1,3:1,5:1,7:1,9:1, compounding dose of institute is for indoor electric physiology and performance testing;
(2) feeler process and EAG test program
The each chemical reagent sample of 10ul is extracted with microsyringe, be added drop-wise to (25mm × 8mm) on doubling filter paper, with tweezers, filter paper is put into Pasteur pipe after candidate agent volatilization 5min, ParafilmTM Pasteur pipe two ends, sample volatilizees after 5min and measures in pipe, it is 500ms that the phase is gone through in stimulation, stimulus intervals is about 30s, the flow velocity of continuous flow is all 4mL/s with stimulating the flow velocity of air-flow, every cephalont only takes 1 feeler, and 6 feelers are tested in each process, and every root feeler repeats 3 times, contrast hockets with the mensuration of processed group, take methyl alcohol as blank.
3. the Holotrichia parallela gyplure of cis-3-hexene ester synergistic effect according to claim 1 or 2, it is characterized in that, described gyplure: the mixture formed when cis-3-hexene ester is 3:1, Electroantennogram signal (EAG) response value of female worm is 2.35 ± 0.56mV.
4. the Holotrichia parallela gyplure of cis-3-hexene ester synergistic effect according to claim 1 or 2, it is characterized in that, described gyplure: the mixture formed when cis-3-hexene ester is 5:1, Electroantennogram signal (EAG) response value of male worm is 3.32 ± 0.55mV.
5. the olfaction reaction method of testing of the Holotrichia parallela gyplure of cis-3-hexene ester synergistic effect according to claim 1, is characterized in that, specifically comprise the steps:
(1) preparation of stimulated samples
The preparation of plant volatiles single dose: each compound all uses methanol as solvent, is configured to 0.1,1,10 μ g/ μ l, 3 concentration, compare with methyl alcohol;
The preparation of plant volatiles and gyplure mixture: pheromones component ILE methyl esters, (R)-(-)-linalool and 1-cyclohexane-carboxylic acid are that 6:1:0.1 ratio is hybridly prepared into Holotrichia parallela gyplure according to weight proportion by this patent, with the mother liquor of 1 μ g/ μ l, according to gyplure: the ratio mixing of plant volatiles=1:1,3:1,5:1,7:1,9:1, compounding dose of institute is for indoor electric physiology and performance testing;
(2) Holotrichia parallela is measured to the susceptibility size of various reagent by " Y " type olfactometer, olfactometer is placed in darkroom, room temperature 27 DEG C, air velocity 500mL/min, get each reagent dropwise of 10ul in test on filter paper bar, and put into " Y " type pipe side arm air inlet place, methyl alcohol contrasts, often group exchanges experimental group and control group seat after repeating, and avoids experimental error; The filter paper bar renewed after each test, often kind of reagent repeats for 3 times, and rear replacing " Y " type pipe surveyed by reagent of the same race, with the effective acetone cleaning of " Y " type crossed, 60 DEG C of oven dry; Often group test picking 10 was unearthed and active strong examination worm the same day, put into " Y " type pipe principal arm port, observed examination worm distribution situation after 10min, and calculate reactivity, relative response rate, reaction coefficient carry out data analysis;
Reactivity=[(in taste source capsule in borer population+control tube borer population)/test total borer population];
Select reactivity=[borer population in taste source capsule/(in taste source capsule in borer population+control tube borer population)].
6. the Holotrichia parallela gyplure of cis-3-hexene ester synergistic effect according to claim 1 or 5, it is characterized in that, described gyplure: the mixture formed when cis-3-hexene ester is 3:1, the selection reactivity of female worm is the highest, is 0.90 ± 0.01.
7. the Holotrichia parallela gyplure of cis-3-hexene ester synergistic effect according to claim 1 or 5, it is characterized in that, described gyplure: the mixture formed when cis-3-hexene ester is 3:1, the selection reactivity of male worm is the highest, is 0.93 ± 0.07.
8. the field of the Holotrichia parallela gyplure of cis-3-hexene ester synergistic effect lures worm test method, it is characterized in that, specifically comprise the steps: to select field trial base, every 60m arranges a trapper, described trapper is by top cover, funnel type device, intersection baffle plate and gathering unit composition, the baffle plate that wherein intersects is located at below top cover, funnel device is located at and is intersected between baffle plate, gathering unit is located at below funnel device, the suspension height of trapper (top) is 2.0m, with the mother liquor of 360ug/ul, gyplure and acetic acid trans-2-hexene ester are mixed with 1:1, 3:1, 5:1, 7:1, 9:1 five different proportions lure core, the described core carrier main component that lures is absorbent cotton, concentration is that 360mg/ lures core, core is lured to concentrate freezen protective at refrigerator, respectively with plant volatiles matter single dose and gyplure as a control group, often lure the repetition of three, core every day, choose five unearthed day data analysis, after trapping every day terminates, record each trapper and lure borer population amount, calculating lures core to the synergistic effect of gyplure, computational methods are as follows:
9. the Holotrichia parallela gyplure after cis-3-hexene ester synergy according to claim 1 or 8, it is characterized in that, described gyplure: during cis-3-hexene ester=5:1, often luring core to lure female borer population to reach maximum every day is 11.00 ± 0.76, be significantly higher than 2.67 ± 0.33 of gyplure, the synergistic effect of cis-3-hexene ester to gyplure reaches 311%.
10. the Holotrichia parallela gyplure after cis-3-hexene ester synergy according to claim 1 or 8, it is characterized in that, described gyplure: during cis-3-hexene ester=3:1, often luring core to lure male worm number to reach maximum every day is 285.00 ± 8.44, be significantly higher than 116 ± 2.65 of gyplure, the synergistic effect of cis-3-hexene ester to gyplure reaches 146%.
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| CN105410038A (en) * | 2015-11-17 | 2016-03-23 | 山东省花生研究所 | Holotrichia parallela sex pheromone synergist and preparation method |
| CN105410038B (en) * | 2015-11-17 | 2018-04-10 | 山东省花生研究所 | A kind of Holotrichia parallela sex pheromone synergist and preparation method |
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| CN110881464A (en) * | 2019-12-05 | 2020-03-17 | 中国农业科学院植物保护研究所 | Attractant formula for preventing and treating Holotrichia parallela imagoes in North China |
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