CN108184906A - A kind of preparation method of attractive insecticide for fruit fly - Google Patents
A kind of preparation method of attractive insecticide for fruit fly Download PDFInfo
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- CN108184906A CN108184906A CN201711491865.2A CN201711491865A CN108184906A CN 108184906 A CN108184906 A CN 108184906A CN 201711491865 A CN201711491865 A CN 201711491865A CN 108184906 A CN108184906 A CN 108184906A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
Abstract
The present invention relates to technical field of pesticide, and in particular to a kind of preparation method of attractive insecticide for fruit fly.The present invention is using ethyl acetoacetate and ethyl chloroacetate as raw material; enzymatically reduction reaction has synthesized product as sex hormones in a kind of and drosophila male worm body; Bake saccharomycete fertility is strong; expression activitiy is high; enzymatic reaction is carried out in this case; product purity, the activity of gained are high; extraction efficiency also improves therewith; by the diverging of drosophila sex pheromone, attract different in nature drosophila aggregation, effective protection and utilize natural enemy; reduce pollution, environmental protection; to killing drosophila group is concentrated to have the effect of fine, killing radix is big, and control effect is good；Sex attractant and pathogenic microorganism bacterium are combined by the present invention, first drosophila are assembled, then drosophila is infected by biological control strain, it kills, no chemical residues in the process will not cause secondary pollution, so as to achieve the effect that the biological control of drosophila and control.
Description
Technical field
The present invention relates to technical field of pesticide, and in particular to a kind of preparation method of attractive insecticide for fruit fly.
Background technology
At present, novel agricultural is in state of development, and the type and area in orchard are constantly expanding, however drosophila is fruit tree
Pest, its life habit so that it is distributed in the entire torrid zone and subtropical zone, and fruit tree just plant with these
Area always has most fruit to be tortured by drosophila in annual orchard, orchard worker is made to suffer heavy losses.
Drosophila Diptera Tephritidae small insect.It is distributed in different continents, endangers fruit and vegetable, it is known that kind 253
It is a, countries in the world fruits and vegetables agriculture is made to suffer huge economic losses, sick disease is propagated in African drosophila, endangers human body health.
Appearance of the drosophila in south China each province causes the great attention of country.Drosophila as common pest in agricultural,
It needs to kill it in many instances.Use the chemical gyplure of male and chemicals poisoning drosophila both at home and abroad at present,
Control disaster.West largely uses " horse traction is fragrant " nerve poisoning agent, though insecticidal power is good, has to people, animal, Pollinating Insect
Evil, influences fruits and vegetables yield." horse traction is fragrant " makes one to vomit, has a headache, has difficulty in breathing, and is detrimental to health.The change that China uses at present
It learns to farm medicine, there are pollution environment, pesticide residue such as is harmful to animal and Pollinating Insect at the drawbacks.In addition, many agricultures in the prior art
Medicine can be killed by sprinkling.But it is time-consuming and laborious to spray insecticide, and pesticide is easily remained in fruit, trees, to it
Later produces, eats and cause unnecessary influence.Pest control with insecticide ingredient, biological degradability is commonly used in this insecticides
Difference causes it to pollute the environment, and its effective time is shorter.
In conclusion existing attractive insecticide for fruit fly is primarily present the defects of following：Essentially synthetics, effective time
Short, biodegradability is poor, easily causes the harm of pesticide residue, and can cause secondary pollution to human body and environment.
Therefore, we it is necessary to again prepare a kind of novel attractive insecticide for fruit fly, present fruit kind is solved with this
Plant industry present in drosophila problem, with this come preferably be adapted to society with the demand in relation to industry, this will be one very
Significant thing.
Invention content
The technical problems to be solved by the invention：For current existing attractive insecticide for fruit fly, essentially synthetics, have
The effect time is short, and biodegradability is poor, easily causes the harm of pesticide residue, and secondary pollution can be caused to ask human body and environment
Topic, provides a kind of preparation method of attractive insecticide for fruit fly.
In order to solve the above technical problems, the present invention is using technical solution as described below：
A kind of preparation method of attractive insecticide for fruit fly, the preparation method include the following steps：
（1）Take Bake yeast in mass ratio 1：It is cultivated in 5 addition yeast culture mediums, takes culture in mass ratio 10：5：2 add in second
Ethyl acetoacetic acid ethyl ester and ethyl chloroacetate, hybrid reaction, centrifugation take filter residue, filter residue in mass ratio 1：8 add in ethyl acetate mixing,
It is filtered through kieselguhr adsorption layer, takes filtrate in mass ratio 1：10 add in sodium chloride, are extracted with ethyl acetate, take extract quality
Score is washed for 26% sodium chloride solution, rotary evaporation, dry, obtains matrix object；
（2）Take matrix object in mass ratio 1：4：1 adds in dihydropyran and lithium aluminium hydride reduction mixing, and reaction obtains reactant A, and negating should
Object A is chromatographed, and takes chromatography object in mass ratio 3：1 adds in paratoluensulfonyl chloride mixing, obtains mixture, takes magnesium powder in mass ratio 1：12~
15：1 adds in tetrahydrofuran and bromo n -nonane, mixes 1 ~ 2h in -15 DEG C, obtains mixture, according to the mass fraction, take 10 ~ 12 parts
Mixture, 5 ~ 6 parts of matrix objects, 2 ~ 3 parts of tetrahydrofurans, 20 ~ 30 parts of aqueous ammonium chloride solution mixing are stirred to react, obtain reactant B,
It is extracted with ether, takes extract liquor rotary evaporation, obtain rotary evaporation object, is i.e. property derivant；
（3）Soil sampling in mass ratio 1：5 addition sterile water mixing are placed in illumination box, and the drosophila larvae of artificial feeding is placed in
In soil, preservative film sealing, preservative film uniformly pricks 10 ~ 15, hole, in 25 ~ 28 DEG C, holding soil moisture 50 ~ 60%, raising children
Worm observes the survival condition of drosophila larvae day by day, collects dead drosophila larvae in mass ratio 1：6 add in sterile water mixing, are homogenized,
Homogenate is taken to be coated in isolation and purification culture base to cultivate, the bacterium colony of picking bacterium diameter maximum, streak inoculation to isolation and purification culture
It is cultivated in base, repeats scribing line culture 2 ~ 3 times, obtain purifying bacterium colony, take purifying bacterium colony in mass ratio 1：9：5 add in physiological saline and matter
It measures score to mix for 0.05% Tween-80, bacteria suspension is made, takes in bacterial suspension inoculation to fermentation medium, fermented and cultured takes hair
Zymotic fluid centrifuges, and takes supernatant, obtains insecticide, takes insecticide and property derivant in mass ratio 2：5 are uniformly mixed, and a lures up to drosophila
Kill agent.
The step（1）Middle yeast culture medium is according to the mass fraction, takes 1 ~ 2 part of NaH2PO4, 0.8 ~ 1.2 part of (NH4)2SO4, 0.2 ~ 0.4 part of MgSO4·7H2O, 0.3 ~ 0.5 part of CaCl2·2H2O, 1 ~ 2 part of yeast extract, 15 ~ 30 parts of glucose, 1 ~ 2 part
Peptone, 500 parts of water mixing, 121 DEG C of sterilizing 30min.
The step（1）Middle Bake Yeast Cultivation condition for 28 ~ 30 DEG C culture 18 ~ for 24 hours；Reaction condition is：25 ~ 28 DEG C anti-
Answer 10 ~ 15h.
The step（2）Reactant A reaction condition is：23 ~ 25 DEG C of 1 ~ 2h of reaction；Reactant B reaction condition is：1~4℃
It is stirred to react 12 ~ 15h.
The step（3）Middle isolation and purification culture base：According to the mass fraction, 180 ~ 200 portions of potatos, 20 ~ 25 parts of Portugals are taken
Grape sugar, 20 ~ 23 parts of agar, 1000 parts of distilled water, 121 DEG C of sterilizing 15min；Fermentation medium：According to the mass fraction, 10 ~ 15 are taken
Part corn flour, 25 ~ 30 parts of wheat bran, 12 ~ 14 parts of rice flour, 20 ~ 25 parts of husks, 300 parts of water, 121 DEG C of sterilizing 15min.
The step（3）The drosophila larvae of middle artificial feeding：Rearing conditions for 28 ~ 30 DEG C of temperature, relative humidity 40 ~ 70%,
Photoperiod L/D=16h/8h；Soil collection：The soil under red bayberry fruit tree is taken, removes table soil, chooses the soil in 20cm below surface layer
Earth crosses 80 mesh and weeds out stone sundries, is sufficiently mixed, is sealed in 4 DEG C.
The step（3）In be separately cultured condition：23 ~ 28 DEG C of 3 ~ 5d of culture；Purifying condition of culture is：25 ~ 28 DEG C of cultures 2
~3d；The inoculum concentration of bacteria suspension is 10%, fermentation culture conditions：30 ~ 35 DEG C of 2 ~ 3d of fermented and cultured.
Compared with other methods, advantageous effects are the present invention：
（1）The present invention is using ethyl acetoacetate and ethyl chloroacetate as raw material, and enzymatically reduction reaction has synthesized a kind of and drosophila
Product as sex hormones in male worm body, Bake saccharomycete fertility is strong, and expression activitiy is high, and it is anti-to carry out enzymatic in this case
Should, product purity, the activity of gained are high, and extraction efficiency also improves therewith, by the diverging of drosophila sex pheromone, attract different in nature fruit
Fly assembles, effective protection and using natural enemy, reduces pollution, environmental protection, kills drosophila group to concentrating and has the effect of fine, kills
The radix that goes out is big, and control effect is good；
（2）The present invention likes the soil of residential location by acquiring drosophila, by the way that the drosophila larvae of raising is placed in soil, receives
Collection is caught an illness, dead drosophila residuum is beaten, and isolating and purifying to obtain one plant has drosophila the bacterial strain of highly pathogenicity, then pass through expansion
Fermented and cultured, enhances the activity for obtaining bacterial strain, which has stronger decomposition host insect epidermis
Enzymatic activity, at the initial stage of infecting, spore surface can generate special protein and enzyme, this proteinoid and enzyme energy and carbohydrate
It is combined, so as to become easier to the combination between spore and host, the germ tube of spore germination president under appropriate conditions
Go out a kind of appresorium with stickum, spore is enable further to be specifically immobilized to Cuticle, stretch out intrusion later
Thorn, drills out an aperture by epicuticle, invades exocuticle, the apex for invading thorn is enlarged into along the direction parallel with epidermis
Tabular, into mycelioid, by the effect of physical mechanical pressure, mycelia gradually invades endepidermis, finally reaches true for end elongation
The process that cortex entomogenous fungi penetrates Cuticle is the synergy of mechanical pressure and enzyme, after intrusion structure penetrates body wall, is led to
The growth for crossing mycelia extends and forms mycelia into haemocoele, generation hyphal body or extension, when germ proliferation is more than certain amount,
Drosophila resistance is no longer valid, and mycelia is constantly proliferated, and gradually invades each internal, finally full of body cavity, leads to the dead of drosophila
It dies；
（3）Sex attractant and pathogenic microorganism bacterium are combined by the present invention, first drosophila are assembled, then pass through biological control strain
Drosophila is infected, is killed, no chemical residues, will not cause secondary pollution in the process, so as to which the biology for having reached drosophila is prevented
The effect controlled and controlled.
Specific embodiment
Bake yeast：It is purchased from Sigam companies.
The drosophila larvae of artificial feeding：Rearing conditions are：28 ~ 30 DEG C of temperature, relative humidity 40 ~ 70%, photoperiod L/D=
16h/8h。
Soil collection：The soil under red bayberry fruit tree is taken, removes table soil, the soil in 20cm below surface layer is chosen, crosses 80 mesh
Stone sundries is weeded out, is sufficiently mixed, is sealed in 4 DEG C.
Isolation and purification culture base：According to the mass fraction, 180 ~ 200 portions of potatos, 20 ~ 25 parts of glucose, 20 ~ 23 parts of fine jades are taken
Fat, 1000 parts of distilled water, 121 DEG C of sterilizing 15min.
Yeast culture medium：According to the mass fraction, 1 ~ 2 part of NaH is taken2PO4, 0.8 ~ 1.2 part of (NH4)2SO4, 0.2 ~ 0.4 part
MgSO4·7H2O, 0.3 ~ 0.5 part of CaCl2·2H2O, 1 ~ 2 part of yeast extract, 15 ~ 30 parts of glucose, 1 ~ 2 part of peptone, 500 parts of water
Mixing, 121 DEG C of sterilizing 30min.
Fermentation medium：According to the mass fraction, 10 ~ 15 portions of corn flour are taken, 25 ~ 30 parts of wheat bran, 12 ~ 14 parts of rice flour, 20 ~ 25
Part husk, 300 parts of water, 121 DEG C of sterilizing 15min.
A kind of preparation method of attractive insecticide for fruit fly, includes the following steps：
（1）Take Bake yeast in mass ratio 1：5 add in yeast culture mediums, in 28 ~ 30 DEG C cultivate 18 ~ for 24 hours, take culture by quality
Than 10：5：2 add in ethyl acetoacetate and ethyl chloroacetate, and 10 ~ 15h is reacted in 25 ~ 28 DEG C, and centrifugation takes filter residue, and filter residue presses matter
Measure ratio 1：8 add in ethyl acetate mixing, are filtered through kieselguhr adsorption layer, take filtrate in mass ratio 1：10 add in sodium chloride, use second
Acetoacetic ester extracts, and extract is taken to be washed with mass fraction for 26% sodium chloride solution, and dry, rotary evaporation obtains matrix object；
（2）Take matrix object in mass ratio 1：4：1 adds in dihydropyran and lithium aluminium hydride reduction mixing, reacts 1 ~ 2h in 23 ~ 25 DEG C, obtains instead
Object A is answered, reactant A is taken to chromatograph, takes chromatography object in mass ratio 3：1 add in paratoluensulfonyl chloride mixing, obtain mixture, take magnesium powder by
Mass ratio 1：12~15：1 adds in tetrahydrofuran and bromo n -nonane, mixes 1 ~ 2h in -15 DEG C, mixture is obtained, by mass fraction
Meter takes 10 ~ 12 parts of mixtures, 5 ~ 6 parts of matrix objects, 2 ~ 3 parts of tetrahydrofurans, 20 ~ 30 parts of aqueous ammonium chloride solution mixing, in 1 ~ 4 DEG C
12 ~ 15h is stirred to react, obtains reactant B, is extracted with ether, takes extract liquor rotary evaporation, obtains rotary evaporation object, is i.e. property derivant；
（3）Soil sampling in mass ratio 1：5 addition sterile water mixing are placed in illumination box, and the drosophila larvae of artificial feeding is placed in
In soil, preservative film sealing, preservative film uniformly pricks 10 ~ 15, hole, in 25 ~ 28 DEG C, holding soil moisture 50 ~ 60%, raising children
Worm observes the survival condition of drosophila larvae day by day, collects dead drosophila larvae in mass ratio 1：6 add in sterile water mixing, are homogenized,
Homogenate is taken to be coated in isolation and purification culture base, 3 ~ 5d, the bacterium colony of picking bacterium diameter maximum, streak inoculation are cultivated in 23 ~ 28 DEG C
Into isolation and purification culture base, 25 ~ 28 DEG C culture 2 ~ 3d, repeat scribing line culture 2 ~ 3 times, obtain purifying bacterium colony, take purifying bacterium colony by
Mass ratio 1：9：5 add in physiological saline and mass fraction mixes for 0.05% Tween-80, and bacteria suspension is made, takes bacteria suspension by 10%
Inoculum concentration be seeded in fermentation medium, in 30 ~ 35 DEG C of 2 ~ 3d of fermented and cultured, zymotic fluid is taken to centrifuge, takes supernatant, obtains desinsection
Agent takes insecticide and property derivant in mass ratio 2：5 are uniformly mixed to get attractive insecticide for fruit fly.
Embodiment 1
Bake yeast：It is purchased from Sigam companies.
The drosophila larvae of artificial feeding：Rearing conditions are：28 DEG C of temperature, relative humidity 40%, photoperiod L/D=16h/8h.
Soil collection：The soil under red bayberry fruit tree is taken, removes table soil, the soil in 20cm below surface layer is chosen, crosses 80 mesh
Stone sundries is weeded out, is sufficiently mixed, is sealed in 4 DEG C.
Isolation and purification culture base：According to the mass fraction, 180 portions of potatos are taken, 20 parts of glucose, 20 parts of agar, 1000 parts
Distilled water, 121 DEG C of sterilizing 15min.
Yeast culture medium：According to the mass fraction, 1 part of NaH is taken2PO4, 0.8 part of (NH4)2SO4, 0.2 part of MgSO4·7H2O,
0.3 part of CaCl2·2H2O, 1 part of yeast extract, 15 parts of glucose, 1 part of peptone, 500 parts of water mixing, 121 DEG C of sterilizing 30min.
Fermentation medium：According to the mass fraction, 10 portions of corn flour are taken, 25 parts of wheat bran, 12 parts of rice flour, 20 parts of husks, 300 parts
Water, 121 DEG C of sterilizing 15min.
A kind of preparation method of attractive insecticide for fruit fly, includes the following steps：
（1）Take Bake yeast in mass ratio 1：5 add in yeast culture medium, cultivate 18h in 28 DEG C, take culture in mass ratio 10：
5：2 add in ethyl acetoacetate and ethyl chloroacetate, and 10h is reacted in 25 DEG C, and centrifugation takes filter residue, filter residue in mass ratio 1：8 add in
Ethyl acetate mixes, and is filtered through kieselguhr adsorption layer, takes filtrate in mass ratio 1：10 add in sodium chloride, are extracted with ethyl acetate,
Extract is taken to be washed with mass fraction for 26% sodium chloride solution, dry, rotary evaporation obtains matrix object；
（2）Take matrix object in mass ratio 1：4：1 adds in dihydropyran and lithium aluminium hydride reduction mixing, reacts 1h in 23 DEG C, obtains reactant
A takes reactant A to chromatograph, and takes chromatography object in mass ratio 3：1 adds in paratoluensulfonyl chloride mixing, obtains mixture, takes magnesium powder by quality
Than 1：12：1 adds in tetrahydrofuran and bromo n -nonane, mixes 1h in -15 DEG C, obtains mixture, according to the mass fraction, takes 10 parts to mix
Conjunction object, 5 parts of matrix objects, 2 parts of tetrahydrofurans, 20 parts of aqueous ammonium chloride solutions mixing are stirred to react 12h in 1 DEG C, obtain reactant B, use
Ether extracts, and takes extract liquor rotary evaporation, obtains rotary evaporation object, is i.e. property derivant；
（3）Soil sampling in mass ratio 1：5 addition sterile water mixing are placed in illumination box, and the drosophila larvae of artificial feeding is placed in
In soil, preservative film sealing, preservative film uniformly pricks 10, hole, and in 25 DEG C, holding soil moisture 50%, breeding grub is observed day by day
The survival condition of drosophila larvae collects dead drosophila larvae in mass ratio 1：6 add in sterile water mixing, and homogenate takes homogenate to apply
It is distributed in isolation and purification culture base, 3d, the bacterium colony of picking bacterium diameter maximum, streak inoculation to isolation and purification culture base is cultivated in 23 DEG C
In, 25 DEG C of culture 2d repeat scribing line culture 2 times, obtain purifying bacterium colony, take purifying bacterium colony in mass ratio 1：9：5 add in physiological saline
It is mixed with mass fraction for 0.05% Tween-80, bacteria suspension is made, bacteria suspension is taken to be seeded to fermentation medium by 10% inoculum concentration
In, in 30 DEG C of fermented and cultured 2d, zymotic fluid is taken to centrifuge, takes supernatant, obtains insecticide, take insecticide and property derivant in mass ratio
2：5 are uniformly mixed to get attractive insecticide for fruit fly.
Embodiment 2
Bake yeast：It is purchased from Sigam companies.
The drosophila larvae of artificial feeding：Rearing conditions are：29 DEG C of temperature, relative humidity 50%, photoperiod L/D=16h/8h.
Soil collection：The soil under red bayberry fruit tree is taken, removes table soil, the soil in 20cm below surface layer is chosen, crosses 80 mesh
Stone sundries is weeded out, is sufficiently mixed, is sealed in 4 DEG C.
Isolation and purification culture base：According to the mass fraction, 190 portions of potatos are taken, 22 parts of glucose, 22 parts of agar, 1000 parts
Distilled water, 121 DEG C of sterilizing 15min.
Yeast culture medium：According to the mass fraction, 1 part of NaH is taken2PO4, 1.0 parts of (NH4)2SO4, 0.3 part of MgSO4·7H2O,
0.5 part of CaCl2·2H2O, 1 part of yeast extract, 20 parts of glucose, 1 part of peptone, 500 parts of water mixing, 121 DEG C of sterilizing 30min.
Fermentation medium：According to the mass fraction, 12 portions of corn flour are taken, 27 parts of wheat bran, 13 parts of rice flour, 23 parts of husks, 300 parts
Water, 121 DEG C of sterilizing 15min.
A kind of preparation method of attractive insecticide for fruit fly, includes the following steps：
（1）Take Bake yeast in mass ratio 1：5 add in yeast culture medium, cultivate 20h in 29 DEG C, take culture in mass ratio 10：
5：2 add in ethyl acetoacetate and ethyl chloroacetate, and 13h is reacted in 27 DEG C, and centrifugation takes filter residue, filter residue in mass ratio 1：8 add in
Ethyl acetate mixes, and is filtered through kieselguhr adsorption layer, takes filtrate in mass ratio 1：10 add in sodium chloride, are extracted with ethyl acetate,
Extract is taken to be washed with mass fraction for 26% sodium chloride solution, dry, rotary evaporation obtains matrix object；
（2）Take matrix object in mass ratio 1：4：1 adds in dihydropyran and lithium aluminium hydride reduction mixing, reacts 1h in 24 DEG C, obtains reactant
A takes reactant A to chromatograph, and takes chromatography object in mass ratio 3：1 adds in paratoluensulfonyl chloride mixing, obtains mixture, takes magnesium powder by quality
Than 1：13：1 adds in tetrahydrofuran and bromo n -nonane, mixes 1h in -15 DEG C, obtains mixture, according to the mass fraction, takes 11 parts to mix
Conjunction object, 5 parts of matrix objects, 2 parts of tetrahydrofurans, 25 parts of aqueous ammonium chloride solutions mixing are stirred to react 13h in 3 DEG C, obtain reactant B, use
Ether extracts, and takes extract liquor rotary evaporation, obtains rotary evaporation object, is i.e. property derivant；
（3）Soil sampling in mass ratio 1：5 addition sterile water mixing are placed in illumination box, and the drosophila larvae of artificial feeding is placed in
In soil, preservative film sealing, preservative film uniformly pricks 12, hole, and in 27 DEG C, holding soil moisture 55%, breeding grub is observed day by day
The survival condition of drosophila larvae collects dead drosophila larvae in mass ratio 1：6 add in sterile water mixing, and homogenate takes homogenate to apply
It is distributed in isolation and purification culture base, 4d, the bacterium colony of picking bacterium diameter maximum, streak inoculation to isolation and purification culture base is cultivated in 24 DEG C
In, 27 DEG C of culture 2d repeat scribing line culture 2 times, obtain purifying bacterium colony, take purifying bacterium colony in mass ratio 1：9：5 add in physiological saline
It is mixed with mass fraction for 0.05% Tween-80, bacteria suspension is made, bacteria suspension is taken to be seeded to fermentation medium by 10% inoculum concentration
In, in 33 DEG C of fermented and cultured 2d, zymotic fluid is taken to centrifuge, takes supernatant, obtains insecticide, take insecticide and property derivant in mass ratio
2：5 are uniformly mixed to get attractive insecticide for fruit fly.
Embodiment 3
Bake yeast：It is purchased from Sigam companies.
The drosophila larvae of artificial feeding：Rearing conditions are：30 DEG C of temperature, relative humidity 70%, photoperiod L/D=16h/8h.
Soil collection：The soil under red bayberry fruit tree is taken, removes table soil, the soil in 20cm below surface layer is chosen, crosses 80 mesh
Stone sundries is weeded out, is sufficiently mixed, is sealed in 4 DEG C.
Isolation and purification culture base：According to the mass fraction, 200 portions of potatos are taken, 25 parts of glucose, 23 parts of agar, 1000 parts
Distilled water, 121 DEG C of sterilizing 15min.
Yeast culture medium：According to the mass fraction, 2 parts of NaH are taken2PO4, 1.2 parts of (NH4)2SO4, 0.4 part of MgSO4·7H2O,
0.5 part of CaCl2·2H2O, 2 parts of yeast extracts, 30 parts of glucose, 2 parts of peptones, 500 parts of water mixing, 121 DEG C of sterilizing 30min.
Fermentation medium：According to the mass fraction, 15 portions of corn flour are taken, 30 parts of wheat bran, 14 parts of rice flour, 25 parts of husks, 300 parts
Water, 121 DEG C of sterilizing 15min.
A kind of preparation method of attractive insecticide for fruit fly, includes the following steps：
（1）Take Bake yeast in mass ratio 1：5 add in yeast culture medium, in 30 DEG C of cultures for 24 hours, take culture in mass ratio 10：
5：2 add in ethyl acetoacetate and ethyl chloroacetate, and 15h is reacted in 28 DEG C, and centrifugation takes filter residue, filter residue in mass ratio 1：8 add in
Ethyl acetate mixes, and is filtered through kieselguhr adsorption layer, takes filtrate in mass ratio 1：10 add in sodium chloride, are extracted with ethyl acetate,
Extract is taken to be washed with mass fraction for 26% sodium chloride solution, dry, rotary evaporation obtains matrix object；
（2）Take matrix object in mass ratio 1：4：1 adds in dihydropyran and lithium aluminium hydride reduction mixing, reacts 2h in 25 DEG C, obtains reactant
A takes reactant A to chromatograph, and takes chromatography object in mass ratio 3：1 adds in paratoluensulfonyl chloride mixing, obtains mixture, takes magnesium powder by quality
Than 1：15：1 adds in tetrahydrofuran and bromo n -nonane, mixes 2h in -15 DEG C, obtains mixture, according to the mass fraction, takes 12 parts to mix
Conjunction object, 6 parts of matrix objects, 3 parts of tetrahydrofurans, 30 parts of aqueous ammonium chloride solutions mixing are stirred to react 15h in 4 DEG C, obtain reactant B, use
Ether extracts, and takes extract liquor rotary evaporation, obtains rotary evaporation object, is i.e. property derivant；
（3）Soil sampling in mass ratio 1：5 addition sterile water mixing are placed in illumination box, and the drosophila larvae of artificial feeding is placed in
In soil, preservative film sealing, preservative film uniformly pricks 15, hole, and in 28 DEG C, holding soil moisture 60%, breeding grub is observed day by day
The survival condition of drosophila larvae collects dead drosophila larvae in mass ratio 1：6 add in sterile water mixing, and homogenate takes homogenate to apply
It is distributed in isolation and purification culture base, 5d, the bacterium colony of picking bacterium diameter maximum, streak inoculation to isolation and purification culture base is cultivated in 28 DEG C
In, 28 DEG C of culture 3d repeat scribing line culture 3 times, obtain purifying bacterium colony, take purifying bacterium colony in mass ratio 1：9：5 add in physiological saline
It is mixed with mass fraction for 0.05% Tween-80, bacteria suspension is made, bacteria suspension is taken to be seeded to fermentation medium by 10% inoculum concentration
In, in 35 DEG C of fermented and cultured 3d, zymotic fluid is taken to centrifuge, takes supernatant, obtains insecticide, take insecticide and property derivant in mass ratio
2：5 are uniformly mixed to get attractive insecticide for fruit fly.
Comparative example：The attractive insecticide for fruit fly of Guangzhou bio tech ltd production
Method：Select one piece of 20 mu of big peach garden, be classified as four groups, using as embodiment with comparative example prepared by drosophila lure
Kill agent.Detect the control effect of interior drosophila for 24 hours after medication.
The specific detection case of attractive insecticide for fruit fly such as table 1
Table 1
From the foregoing, it will be observed that the attractive insecticide for fruit fly prepared by the present invention can effectively lure drosophila, and gone out and removed, and the present invention
Component used in attractive insecticide for fruit fly is safe and non-stimulating ingredient, in addition, the present invention preparation method it is simple to operation and
It is of low cost, it is a kind of safety and the preparation method of efficient attractive insecticide for fruit fly, is worthy to be popularized and uses.
Claims (7)
1. a kind of preparation method of attractive insecticide for fruit fly, which is characterized in that the preparation method includes the following steps：
（1）Take Bake yeast in mass ratio 1：It is cultivated in 5 addition yeast culture mediums, takes culture in mass ratio 10：5：2 add in second
Ethyl acetoacetic acid ethyl ester and ethyl chloroacetate, hybrid reaction, centrifugation take filter residue, filter residue in mass ratio 1：8 add in ethyl acetate mixing,
It is filtered through kieselguhr adsorption layer, takes filtrate in mass ratio 1：10 add in sodium chloride, are extracted with ethyl acetate, take extract quality
Score is washed for 26% sodium chloride solution, rotary evaporation, dry, obtains matrix object；
（2）Take matrix object in mass ratio 1：4：1 adds in dihydropyran and lithium aluminium hydride reduction mixing, and reaction obtains reactant A, and negating should
Object A is chromatographed, and takes chromatography object in mass ratio 3：1 adds in paratoluensulfonyl chloride mixing, obtains mixture, takes magnesium powder in mass ratio 1：12~
15：1 adds in tetrahydrofuran and bromo n -nonane, mixes 1 ~ 2h in -15 DEG C, obtains mixture, according to the mass fraction, take 10 ~ 12 parts
Mixture, 5 ~ 6 parts of matrix objects, 2 ~ 3 parts of tetrahydrofurans, 20 ~ 30 parts of aqueous ammonium chloride solution mixing are stirred to react, obtain reactant B,
It is extracted with ether, takes extract liquor rotary evaporation, obtain rotary evaporation object, is i.e. property derivant；
（3）Soil sampling in mass ratio 1：5 addition sterile water mixing are placed in illumination box, and the drosophila larvae of artificial feeding is placed in
In soil, preservative film sealing, preservative film uniformly pricks 10 ~ 15, hole, in 25 ~ 28 DEG C, holding soil moisture 50 ~ 60%, raising children
Worm observes the survival condition of drosophila larvae day by day, collects dead drosophila larvae in mass ratio 1：6 add in sterile water mixing, are homogenized,
Homogenate is taken to be coated in isolation and purification culture base to cultivate, the bacterium colony of picking bacterium diameter maximum, streak inoculation to isolation and purification culture
It is cultivated in base, repeats scribing line culture 2 ~ 3 times, obtain purifying bacterium colony, take purifying bacterium colony in mass ratio 1：9：5 add in physiological saline and matter
It measures score to mix for 0.05% Tween-80, bacteria suspension is made, takes in bacterial suspension inoculation to fermentation medium, fermented and cultured takes hair
Zymotic fluid centrifuges, and takes supernatant, obtains insecticide, takes insecticide and property derivant in mass ratio 2：5 are uniformly mixed to get drosophila trapping
Agent.
2. the preparation method of attractive insecticide for fruit fly according to claim 1, which is characterized in that the step（1）Middle Yeast Cultivation
Base is according to the mass fraction, takes 1 ~ 2 part of NaH2PO4, 0.8 ~ 1.2 part of (NH4)2SO4, 0.2 ~ 0.4 part of MgSO4·7H2O, 0.3 ~ 0.5
Part CaCl2·2H2O, 1 ~ 2 part of yeast extract, 15 ~ 30 parts of glucose, 1 ~ 2 part of peptone, 500 parts of water mixing, 121 DEG C of sterilizings
30min。
3. the preparation method of attractive insecticide for fruit fly according to claim 1, which is characterized in that the step（1）Middle Bake yeast
Condition of culture for 28 ~ 30 DEG C culture 18 ~ for 24 hours；Reaction condition is：25 ~ 28 DEG C of 10 ~ 15h of reaction.
4. the preparation method of attractive insecticide for fruit fly according to claim 1, which is characterized in that the step（2）Reactant A reacts
Condition is：23 ~ 25 DEG C of 1 ~ 2h of reaction；Reactant B reaction condition is：1 ~ 4 DEG C is stirred to react 12 ~ 15h.
5. the preparation method of attractive insecticide for fruit fly according to claim 1, which is characterized in that the step（3）In isolate and purify
Culture medium：According to the mass fraction, 180 ~ 200 portions of potatos are taken, 20 ~ 25 parts of glucose, 20 ~ 23 parts of agar, 1000 parts of distilled water,
121 DEG C of sterilizing 15min；Fermentation medium：According to the mass fraction, 10 ~ 15 portions of corn flour, 25 ~ 30 parts of wheat bran, 12 ~ 14 portions of rice are taken
Powder, 20 ~ 25 parts of husks, 300 parts of water, 121 DEG C of sterilizing 15min.
6. the preparation method of attractive insecticide for fruit fly according to claim 1, which is characterized in that the step（3）Middle artificial feeding
Drosophila larvae：Rearing conditions are 28 ~ 30 DEG C of temperature, relative humidity 40 ~ 70%, photoperiod L/D=16h/8h；Soil collection：It takes
Soil under red bayberry fruit tree removes table soil, chooses the soil in 20cm below surface layer, crosses 80 mesh and weeds out stone sundries, fully mixed
It closes, is sealed in 4 DEG C.
7. the preparation method of attractive insecticide for fruit fly according to claim 1, which is characterized in that the step（3）In be separately cultured
Condition：23 ~ 28 DEG C of 3 ~ 5d of culture；Purifying condition of culture is：25 ~ 28 DEG C of 2 ~ 3d of culture；The inoculum concentration of bacteria suspension is 10%, fermentation
Condition of culture：30 ~ 35 DEG C of 2 ~ 3d of fermented and cultured.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110122471A (en) * | 2019-05-06 | 2019-08-16 | 泰山医学院 | A kind of flies control method suitable for farm |
| CN115475525A (en) * | 2022-10-06 | 2022-12-16 | 中国矿业大学 | Method for purifying biological dust suppressant by two-stage ultrafiltration |
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| CN102805110A (en) * | 2011-05-31 | 2012-12-05 | 温州医学院 | Fruit fly food source attractant and preparation method thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110122471A (en) * | 2019-05-06 | 2019-08-16 | 泰山医学院 | A kind of flies control method suitable for farm |
| CN110122471B (en) * | 2019-05-06 | 2021-08-17 | 山东第一医科大学(山东省医学科学院) | Fly control method suitable for farm |
| CN115475525A (en) * | 2022-10-06 | 2022-12-16 | 中国矿业大学 | Method for purifying biological dust suppressant by two-stage ultrafiltration |
| CN115475525B (en) * | 2022-10-06 | 2024-03-05 | 中国矿业大学 | Method for purifying biological dust suppressant by double-stage ultrafiltration |
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Application publication date: 20180622 |