CN107917852A - The bioassay method that a kind of noxious material influences green plant bug nymph - Google Patents
The bioassay method that a kind of noxious material influences green plant bug nymph Download PDFInfo
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- CN107917852A CN107917852A CN201711131085.7A CN201711131085A CN107917852A CN 107917852 A CN107917852 A CN 107917852A CN 201711131085 A CN201711131085 A CN 201711131085A CN 107917852 A CN107917852 A CN 107917852A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N5/00—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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Abstract
The present invention provides the bioassay method that a kind of noxious material influences green plant bug nymph, including noxious material to be measured and green plant bug man-made feeds are mixed to prepare experimental group feed, and positive group feed is made with potassium dihydrogen arsenate and/or cystatin；Set blank control group at the same time；Above-mentioned each group feed is wrapped up respectively with the sealed membrane of stretching, and feed capsule of the same size is made；Above-mentioned feed capsule is respectively placed in feeding box, green plant bug nymph is accessed into feeding box, is sealed with gauze, be placed under suitable condition and cultivate；From the death condition or changes of weight of the 0th day start recording green plant bug, influence of the evaluation noxious material to green plant bug.This method to stomach toxicity, systemic action medicament and be dissolved in the chemical substances of man-made feeds and can effectively measure, solve the piercing-sucking mouthparts pests including green plant bug be difficult to directly the noxious material of contact various concentrations, uncertain test green plant bug whether feeding man-made feeds the problems such as.
Description
Technical field
The invention belongs to technical field of plant protection, and specifically, being related to a kind of noxious material influences green plant bug nymph
Bioassay method.
Background technology
Green plant bug (Apolygus lucorum Meyer-D ü r) belongs to Semiptera (Hemiptera) Miridae (Miridae),
It is a kind of common piercing-sucking mouthparts pests, its host range is extensive, can endanger to plant including cotton, corn, fruit tree etc. more than 200 and make
Thing.In China, green plant bug is considered as the minor pests that a kind of occurrence degree is lighter in agricultural production in traditional sense, always
Simultaneous object of controlling as bollworm chemical prevention is controlled, without individually being prevented.From 1997, China started to be commercialized
Bt transgenic cotton is planted, bollworm has obtained effective control, and the usage amount of cotton field chemical pesticide significantly reduces, plus crop-planting
The reasons such as the adjustment of structure, cause the quantity of green plant bug field population generation in recent years to steeply rise, being increasingly becoming influences cotton life
A kind of primary pest of production, and green plant bug popution explosion also results in its harm to jujube, grape, cherry, tea tree etc. also increasingly
Seriously, it has also become influence the significant problem of various crop production.
Currently, the prevention of green plant bug is main or based on chemical prevention, but chemical pesticide be excessively used can cause it is green
A series of problems, such as increase of the fleahopper resistance to the action of a drug, environmental pollution, injury natural enemy, food security, therefore, develop some new preventions
The biological insecticides of green plant bug, albumen, enzyme inhibitor etc. are of great significance.Therefore, effectively evaluating noxious material pair is established
The bioassay system that green plant bug influences, can be not only used for screening exactly to the virose insect killing substance of green plant bug tool, goes back
It can be used for evaluating other materials to security of green plant bug etc..
Green plant bug belongs to sucking pest, if the material of measure by soak, smear etc. can be only present in kidney bean or
The surface of corn, it is difficult to be drawn onto by green plant bug thorn；And whether feeding food also is difficult to determine green plant bug, raw survey system has
Effect property and stability are difficult to keep.
The content of the invention
In order to solve the above technical problems, the present invention provides the bioassay side that a kind of noxious material influences green plant bug nymph
Method, by the method for the present invention, the method not being wrapped in only with low dose of feed in sealed membrane self-control Caplet meets green plant bug
Feeding, and specify that valid density of the positive control substance in raw survey system so that raw survey system has preferable validity
And sensitivity.
Technical solution of the present invention is as follows：
The bioassay method that a kind of noxious material influences green plant bug nymph, includes the following steps：
1) noxious material to be measured is diluted to various concentrations, is mixed with green plant bug man-made feeds, be made and contain Toxic to be measured
The experimental group feed of matter, the green plant bug man-made feeds not add any material are used as blank control group feed；
To add 8 μ g/g potassium dihydrogen arsenates (KH2AsO4, PA) the green plant bug man-made feeds, and/or, to add 4 μ g/
The institute of g cystatins (N- [trans-epoxy succinyl base]-L-Leu -4- guanidinobutyl acid amides, E-64)
It is positive controls feed to state green plant bug man-made feeds；
2) above-mentioned experimental group feed, blank control group feed and positive controls are wrapped up respectively with the sealed membrane of stretching to raise
Material, is made feed capsule of the same size；
3) by above-mentioned feed capsule be respectively placed in feeding box in, to feeding box in access green plant bug nymph (preferably 1 age) if
Worm, is sealed with gauze (preferably 150 mesh), is placed in 27 ± 2 DEG C of temperature, humidity 75 ± 0%, under conditions of illumination 14h, dark 10h
Culture；
4) from the death condition or changes of weight of the 0th day start recording green plant bug, so as to evaluate noxious material to green plant bug
Influence.
Further, the sealed membrane size is 2.5cm × 2.5cm, is drawn into 6cm × 8cm, and 0.4- is added dropwise in film center
0.6mL feeds.
Further, the feeding box is water white transparency beaker, such as diameter 4.0cm, high 6.0cm.
Further, raise box to replace once within every two days in feed capsule, the survival of the nymph of timing observation daily and husking feelings
Condition, after green plant bug nymphal development to adult, the adult newly to sprout wings in each processing is weighed, and identifies female, male and random pair,
It is transferred in the insect box that hydromel is provided, to Adult supplement nutrient, adult survival condition and daily egg laying amount is recorded, until adult
It is dead.
Man-made feeds involved in the present invention are the green plant bug man-made feeds prepared according to ZL201010556195.X.Manually
Feed stores under conditions of 0-4 DEG C, and storage time is no more than 30 days.
Bioassay method provided by the invention, it is applied widely, to stomach toxicity, systemic action medicament and be dissolved in people
The chemical substance of work feed can be measured effectively, solved the piercing-sucking mouthparts pests including green plant bug and be difficult to directly connect
Touch the noxious materials of various concentrations, uncertain test green plant bug whether feeding man-made feeds the problems such as, specify that positive material
In the valid density of raw survey system so that raw survey system has preferable validity and sensitivity, can Accurate Determining noxious material
Influence to green plant bug, has very strong practicality.
Embodiment
Following embodiments are used to illustrate the present invention, but are not limited to the scope of the present invention.It is not specified in embodiment specific
Technology or condition person, carry out according to the described technology of document in the art or condition, or according to product description.It is used
Production firm person is not specified in reagent or instrument, is the conventional products that can be commercially available by regular distributor.
The green plant bug man-made feeds used in following embodiments are according to the method system disclosed in ZL 201010556195.X
It is standby.
Embodiment 1 evaluates influence of the Cry1Ac albumen to green plant bug nymph
Green plant bug man-made feeds 100mL is prepared, puts it into 4 DEG C of refrigerators and stores, it is spare.
Potassium dihydrogen arsenate (KH2AsO4, PA) or cystatin (N- [trans-epoxy succinyl base]-L-
Leucine -4- guanidinobutyl acid amides, E-64) Sigma-Aldrich (St.Louis, MO, USA) is purchased from, it is stored in -20 before use
DEG C refrigerator.
The Cry1Ac albumen of 400 μ g/g is added in man-made feeds, feed to be determined is prepared into, as experimental group；With not
The man-made feeds of other compositions are added as negative control group (CK)；To add 8 μ g/g potassium dihydrogen arsenates (KH2AsO4, PA) institute
State green plant bug man-made feeds, and/or, with add 4 μ g/g cystatins (N- [trans-epoxy succinyl base]-
L-Leu -4- guanidinobutyl acid amides, E-64) the green plant bug man-made feeds be positive controls feed；With the envelope of stretching
Membrana oralis wraps up above-mentioned experimental group feed, blank control group feed and positive controls feed respectively, and feed of the same size is made
Capsule.The sealed membrane size is 2.5cm × 2.5cm, is drawn into 6cm × 8cm, and 0.4-0.6mL feeds are added dropwise in film center.
Experimental group, blank control group (CK), PA positive controls and E-64 positive controls feed capsules are respectively placed in
To raise in box, each raise is respectively connected to 12 first 1 age green plant bug nymphs in box, and with crocus cloth (150 mesh gauze) and rubber ring
Sealing prevents its escape.It is placed in 27 ± 2 DEG C of temperature, humidity 75 ± 10%, is cultivated under conditions of illumination 14h, dark 10h.Feed
Capsule is replaced once for every two days, the survival of the nymph of timing observation daily and husking situation, will after green plant bug nymphal development to adult
The adult newly to sprout wings in each processing is weighed, and identifies female, male and random pair, is transferred in the insect box for providing hydromel, into
Worm supplements the nutrients, and records adult survival condition and daily egg laying amount, until adult is dead.Experiment is repeated 3 times.
The green plant bug life table parameters of different disposal group is shown in Table 1.Table 1 the result shows that：Any other component is not added with feeding
The green plant bug of man-made feeds is compared, and the growth and development parameters of 8 μ g/g PA and 4 μ g/g E-64 to green plant bug nymph have aobvious
Work or pole significant impact, and strong influence is also resulted in egg laying amount after nymph feeding.Show the positive control of selection
Matter is effective and sensitive.And the Cry1Ac albumen parameters and contrast difference measured are not notable, the toxicity to green plant bug is not showed.
The life table parameters of 1 different disposal group green plant bug of table
Note：Test data is analyzed using SPSS (version20), Multiple range test is carried out with Dunnett ' s test detections.
* represent:P<0.05, * * is represented:P<0.01.Listed numerical value is average value ± standard error in table.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (6)
1. the bioassay method that a kind of noxious material influences green plant bug nymph, it is characterised in that include the following steps：
1) noxious material to be measured is diluted to various concentrations, is mixed with green plant bug man-made feeds, is made containing noxious material to be measured
Experimental group feed, the green plant bug man-made feeds not add any material are used as blank control group feed；
To add the green plant bug man-made feeds of 8 μ g/g potassium dihydrogen arsenates, and/or, to add 4 μ g/g cysteine proteinases
The green plant bug man-made feeds of inhibitor are positive controls feed；
2) above-mentioned experimental group feed, blank control group feed and positive controls feed are wrapped up respectively with the sealed membrane of stretching, make
Into feed capsule of the same size；
3) above-mentioned feed capsule is respectively placed in feeding box, accesses green plant bug nymph into feeding box, sealed, be placed in gauze
27 ± 2 DEG C of temperature, humidity 75 ± 0%, is cultivated under conditions of illumination 14h, dark 10h；
4) from the death condition or changes of weight of the 0th day start recording green plant bug, so as to evaluate shadow of the noxious material to green plant bug
Ring.
2. according to the method described in claim 1, it is characterized in that, the sealed membrane size is 2.5cm × 2.5cm, it is drawn into
6cm × 8cm, 0.4-0.6mL feeds are added dropwise in film center.
3. method according to claim 1 or 2, it is characterised in that the feeding box is water white transparency beaker, preferably diameter
4.0cm, high 6.0cm.
4. method according to claim 1 or 2, it is characterised in that the gauze is 150 mesh gauzes.
5. method according to claim 1 or 2, it is characterised in that access 1 age green plant bug nymph into feeding box.
6. according to claim 1-5 any one of them methods, it is characterised in that feed capsule replaces one in every two days in feeding box
Secondary, the survival of the nymph of timing observation daily and husking situation, after green plant bug nymphal development to adult, newly sprout wings in each handling
Adult weigh, identify female, male and random pair, be transferred to provide hydromel insect box in, to Adult supplement nutrient, be recorded as
Worm survival condition and daily egg laying amount, until adult is dead.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113702626A (en) * | 2020-05-20 | 2021-11-26 | 中国农业大学 | Simple, convenient and efficient biological assay reagent and method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113702626A (en) * | 2020-05-20 | 2021-11-26 | 中国农业大学 | Simple, convenient and efficient biological assay reagent and method |
| CN113702626B (en) * | 2020-05-20 | 2023-10-13 | 中国农业大学 | Simple and efficient biological assay reagent and method |
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Application publication date: 20180417 |