CN107907574A - A kind of experimental method of tea geometrid In vivo detection - Google Patents
A kind of experimental method of tea geometrid In vivo detection Download PDFInfo
- Publication number
- CN107907574A CN107907574A CN201710935897.0A CN201710935897A CN107907574A CN 107907574 A CN107907574 A CN 107907574A CN 201710935897 A CN201710935897 A CN 201710935897A CN 107907574 A CN107907574 A CN 107907574A
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- tea geometrid
- adsorption tube
- collecting pipe
- vivo detection
- experimental method
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
Abstract
The invention discloses a kind of method of tea geometrid In vivo detection, including, step 1：Set includes collecting pipe, adsorption tube by pretreated device, and collecting pipe is propped by iron stand, and adsorption tube is by butterfly clamp fixed position, and pulse gas generator is in collecting pipe side.Collecting pipe opposite side is connected with adsorption tube, and adsorption tube is connected with two continuous conical flasks, and the other end is connected with air pump.All connections connect in addition to the connection of adsorption tube and adsorption tube by rubber tube；Step 2：Experiment is divided into experimental group and blank control group, experimental group is put into tea geometrid in collecting pipe, and blank control group is not put then；Step 3：The be connected again electrochemical workstation of sensor-based system of actuating gas generator and air pump is tested, and records data；Step 4：Processing experimental result is simultaneously drawn by the sex pheromone concentration of the female moth release of tea geometrid.This method can be detected effectively by the sex pheromone gas molecule of the female moth live body release of tea geometrid in laboratory environments.
Description
Technical field
The present invention relates to living insects experiment, and in particular to experimental provision and side to tea geometrid In vivo detection pheromones
Method.
Background technology
Tea geometrid is a kind of pest being prevalent in tea place, and serious harm is caused to Tea Production, is caused directly
Economic loss.In the prevention of tea geometrid, main means still find to distinguish and are broken out so as to understand insect pest by human eye at present
Situation.Insect pheromone mainly has trail, alarm pheromones, sex pheromone etc., special using the height of insect pheromone
One property plays the role of monitoring to insect pest situation.The detection for carrying out pheromones to tea geometrid live body in laboratory environment also lacks correlation in fact
Experiment device.The sensor of detection tea geometrid sex pheromone has had relevant application, but related application is simply by the tea of purchase
Looper sex pheromone solution is added drop-wise in sensor, and lacks the detection of the sex pheromone of the female moth live body release of oestrus tea geometrid
Scheme.
The content of the invention
It is an object of the invention to provide a kind of experimental method of tea geometrid In vivo detection.Tea geometrid release property letter can be achieved
Cease the collection of element, detection.
A kind of experimental method of tea geometrid In vivo detection, including：Step 1：Experimental provision, including collecting pipe, absorption are set
Pipe, pulse gas generator, air pump, electrochemical workstation and its auxiliary facility, electrochemical impedance biology sensor etc.；Step 2：
Above-mentioned experimental group is divided into experimental group and blank control；Step 3：Start pulse gas generator air pump, electrochemical biosensor
System carries out tea geometrid sex pheromone test experience, and records data；Step 4：Handle experimental result and environment is calculated
In tea geometrid sex pheromone concentration；
In one embodiment, in step 1, the female moth of tea geometrid should be more than 5 in adsorption tube.
In one embodiment, in step 1 and step 3, the electrode of electrochemica biological sensor selects E-Plate L8
The interdigitated gold electrode of series.
In one embodiment, in step 1 and step 3, electrochemica biological sensor transducer is believed using tea geometrid property
The plain associated proteins 2 of breath（EoblPBP2）Albumen.
In one embodiment, electrochemical impedance biology sensor needs to use in step 1（Z, Z, Z）-3, 6,
18 carbon triolefins of 9- or 6,7- epoxies（Z, Z）18 carbon diene of -3,9- is demarcated.
In one embodiment, in step 4, electrochemical sensing system uses electrochemical impedance spectral technology (EIS).
In one embodiment, collecting pipe is propped by iron stand, and adsorption tube is occurred by butterfly clamp fixed position, pulse gas
Device is in collecting pipe side.Collecting pipe opposite side is connected with adsorption tube, and adsorption tube is connected with two continuous conical flasks, the other end
It is connected with air pump.All connections in addition to the connection of adsorption tube and adsorption tube are connected with rubber tube.
The beneficial effects of the invention are as follows：The tea geometrid biopsy method can be detected effectively by tea in laboratory environments
The sex pheromone of the female moth live body release of looper.
Brief description of the drawings
Fig. 1 is the block schematic illustration that the present invention implements tea geometrid In vivo detection sex pheromone scheme.
Fig. 2 is electrochemical sensor elements preparation figure in the present invention.
Fig. 3 a, 3b are normalized impedance-ligand concentra logarithmic charts.
Fig. 4 is the female moth In vivo detection impedance spectra of tea geometrid.
Fig. 5 A, 5B are normalized impedance-time diagrams, the female moth experimental group 5B. blank control groups of 5A. tea geometrids.
Embodiment
Technical scheme is described in detail below by specific embodiment.
As shown in Figure 1, a kind of experimental assembly and scheme of tea geometrid In vivo detection, in order to realize above-described purpose,
The present invention uses following scheme, and the present apparatus has some sections of sebific ducts, and air pump 1 is slowly pumped into gas in pipeline one end, and gas is through undue
The wide-mouth bottle 3 of activated carbon Zhuan You not be put into the wide-mouth bottle 2 and ultra-pure water of ultra-pure water, this measure clean not moist sky in order to obtain
Gas, the other end insertion adsorption tube 4 of the pipeline, adsorption tube 4 should be directed at certain passage in electrochemical sensing system 5 in experiment.
Another pipeline one end connection pulse gas generator 6, collecting pipe 7 of the other end connection equipped with tea geometrid live body, collecting pipe 7 connect
Onto adsorption tube 4, the sex pheromone that tea geometrid produces is allowed slowly to flow into adsorption tube 4 from collecting pipe 7, then enter through adsorption tube 4
In the sense channel of ready electrochemical sensing system 5.
The gas rate of outflow in collecting pipe 7 is received less times greater than the gas inflow velocity in adsorption tube 4 in order to avoid air enters
Collector 7 and hinder the outflow of smell mixture in collecting pipe；First it is evacuated 10 minutes before collecting pipe connection, removes whole gas flow loop
In possible impurity；Collecting pipe 7 and adsorption tube 4 must be purified before and burin-in process, to dispel wherein impurity；It is different
The tolerance of tea geometrid release is different, and acquisition time is subject to concrete operations；Various glass apparatus and connecting tube will cleanings.
Pretreatment stage is being tested, is being the processing of the female moth of tea geometrid first：Choose non-mating in good condition, that build is healthy and strong
Some of the female moth of tea geometrid, be put into spare in beaker.Followed by filter material is handled：It is all necessary before activated carbon and silica gel experiment
By 120 DEG C of 4 h of processing of high temperature, to exclude moisture therein.Collecting pipe and adsorption tube must be carried out before at purification and aging
Reason, condition flow through collecting pipe and absorption for nitrogen with the flow velocity of 10-50 ml/min, 300-340 DEG C of high temperature, holding 3-6 h
Pipe.
The present invention specific experiment method be：
Step 1：The female moth of tea geometrid five is put into collecting pipe 7 for one group with tweezers, the air inlet 8 of collecting pipe 7 with hose with
Pulse gas generator 6 is connected, and opens the first air circulation of pulse gas generator 6 one minute, fills the gas that tea geometrid discharges
Full collecting pipe 7, to accelerate the progress of subsequent experimental, gas outlet 11 is inserted into the aperture on adsorption tube 4, collecting pipe 7 and absorption
Pipe 4 is fixed with iron stand；The air inlet hose of adsorption tube 4 is with holding the bottle of silica gel and activated carbon and holding ultrapure
The bottle connection of water, for purifying the air into adsorption tube 4, gas outlet aligning electrodes plate hole groove.Collecting pipe inlet port with
Pulse gas generator is connected, and the flowing of gas in enrichment pipe, gas outlet insertion are driven by conveying the air of certain flow rate
In small holes on enrichment pipe, smell is transmitted in electrode plate hole slot by enrichment pipe.The air inlet of adsorption tube 4 respectively with work
Property charcoal bottle, the gentle pump of ultrapure water bottle be connected, the hole slot of gas outlet aligning electrodes sensor-based system.When air pump 1 is opened, air is being inhaled
Certain air stream is formed from air inlet to gas outlet in attached pipe 4.Open pulse gas generator 6, tea geometrid release again at this time
Gas be just fed in adsorption tube 4, and as air-flow flows to electrode plate hole slot together.
Step 2：Experiment is divided into experimental group and blank control group according to the female moth of tea geometrid whether is put into collecting pipe 7.
Step 3：Pulse gas generator 6 and air pump 1 are started, electrochemical workstation uses ac impedance mode, record
The real and imaginary parts data of equivalent circuit impedance.
Step 4：Make the fitting of Zview softwares as detection albumen and the parameter Rct of aglucon interaction.This experimental method
In, electrochemical biosensor system need to first demarcate to obtain NIC-ligand concentra logarithmic curve parameter, i.e.,；
Wherein C is material concentration, and K is curve coefficients, and B is curve intercept,.According to NIC-Log (C)
Curve can converse corresponding tea geometrid release pheromone concentration by corresponding Rct values.
Claims (7)
1. a kind of experimental method of tea geometrid In vivo detection, including：
Step 1：Experimental provision is set, including collecting pipe, adsorption tube, pulse gas generator, air pump, electrochemical workstation and its
Auxiliary facility, electrochemical biosensor system etc.；
Step 2：Above-mentioned experimental group is divided into experimental group and blank control；
Step 3：Pulse gas generator air pump is started, it is real that electrochemical biosensor system carries out tea geometrid sex pheromone detection
Test, and record data；
Step 4：Processing experimental result and the tea geometrid sex pheromone concentration being calculated in environment.
2. the experimental method of tea geometrid In vivo detection according to claim 1, it is characterised in that the adsorption tube in step 1
Using glass-pulling, it is put into easy to the female moth of tea geometrid.
3. the experimental method of tea geometrid In vivo detection according to claim 1, it is characterised in that the collecting pipe in step 1
By glass-pulling, the cleaned air easy to be blasted by air pump is mixed with insect sex pheromone.
4. the experimental method of tea geometrid In vivo detection according to claim 1, it is characterised in that in step 1 in adsorption tube
The female moth of tea geometrid should be more than 5.
5. the experimental method of tea geometrid In vivo detection according to claim 1, it is characterised in that electric in step 1 and step 3
The electrode of chemical biosensor selects the interdigitated gold electrode of E-Plate L8 series.
6. the experimental method of tea geometrid In vivo detection according to claim 1, it is characterised in that electric in step 1 and step 3
The transducer of chemical biosensor uses tea geometrid sex pheromone associated proteins 2（EoblPBP2）.
7. the experimental method of tea geometrid In vivo detection according to claim 1, it is characterised in that electrochemistry passes in step 1
The calibration of sensing system uses（Z, Z, Z）18 carbon triolefins of -3,6,9- or 6,7- epoxies（Z, Z）18 carbon two of -3,9-
Alkene.
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| CN201710935897.0A CN107907574B (en) | 2017-10-10 | 2017-10-10 | Experimental method for tea geometrid living body detection |
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| CN201710935897.0A CN107907574B (en) | 2017-10-10 | 2017-10-10 | Experimental method for tea geometrid living body detection |
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| CN107907574A true CN107907574A (en) | 2018-04-13 |
| CN107907574B CN107907574B (en) | 2020-05-19 |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101234279A (en) * | 2007-11-01 | 2008-08-06 | 浙江大学 | Open type living body plant volatile dynamic high-efficiency collecting device |
| CN103472102A (en) * | 2013-09-29 | 2013-12-25 | 浙江大学 | Preparing method and application of odorant binding protein sensor based on impedance analysis |
| US20160146790A1 (en) * | 2009-03-04 | 2016-05-26 | The Regents Of The University Of California | Insect repellent and attractants |
| CN206057041U (en) * | 2016-09-17 | 2017-03-29 | 中国农业科学院果树研究所 | A kind of Portable field plant volatile collection kit case |
| CN106689133A (en) * | 2016-11-18 | 2017-05-24 | 安徽农业大学 | Tea geometrid sex pheromone composition and lure thereof |
-
2017
- 2017-10-10 CN CN201710935897.0A patent/CN107907574B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101234279A (en) * | 2007-11-01 | 2008-08-06 | 浙江大学 | Open type living body plant volatile dynamic high-efficiency collecting device |
| US20160146790A1 (en) * | 2009-03-04 | 2016-05-26 | The Regents Of The University Of California | Insect repellent and attractants |
| CN103472102A (en) * | 2013-09-29 | 2013-12-25 | 浙江大学 | Preparing method and application of odorant binding protein sensor based on impedance analysis |
| CN206057041U (en) * | 2016-09-17 | 2017-03-29 | 中国农业科学院果树研究所 | A kind of Portable field plant volatile collection kit case |
| CN106689133A (en) * | 2016-11-18 | 2017-05-24 | 安徽农业大学 | Tea geometrid sex pheromone composition and lure thereof |
Non-Patent Citations (3)
| Title |
|---|
| LI HONGLIANG等: "Physicochemical Evidence on Sublethal Neonicotinoid Imidacloprid Interacting with an Odorant-Binding Protein from the Tea Geometrid Moth, Ectropis oblique", 《JOURNAL OF AGRICULTURAL FOOD CHEMISTRY》 * |
| 冯一璐等: "茶尺蠖信息素结合蛋白PBP2的基因克隆、原核表达及其结合功能", 《中国农业科学》 * |
| 刘亚佳等: "菊花瘿蚊雌蚊释放性信息素的确定和提取", 《中国农业大学学报》 * |
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