CN101315360A

CN101315360A - Bioassay method for pest

Info

Publication number: CN101315360A
Application number: CNA2008101171115A
Authority: CN
Inventors: 吴青君; 张治军; 徐宝云; 张友军
Original assignee: Institute of Vegetables and Flowers Chinese Academy of Agricultural Sciences
Current assignee: Institute of Vegetables and Flowers Chinese Academy of Agricultural Sciences
Priority date: 2008-07-24
Filing date: 2008-07-24
Publication date: 2008-12-03
Anticipated expiration: 2028-07-24
Also published as: CN101315360B

Abstract

The invention belongs to the technology field of the plant protection, and particularly relates to a bioassay method for pests. The pest bioassay method is characterized in that a plant tissue which is favored by pests and is processed by medicaments with the same concentration is put in a container with pests therein and medicaments on the inner wall. The method has wide application, effective detections can be carried out to the medicaments with contact killing and stomach poisoning functions at the same time, and the minimum effective dose of each medicament to the pests can be detected accurately, thereby effectively poisoning and killing the pests and minimizing environmental pollution and medicament waste.

Description

The bioassay method of a kind of insect

Technical field

The invention belongs to the plant protection technology field, particularly relate to the bioassay method of a kind of insect

Background technology

Medicament is a method the most efficiently and effectively such as r-countermeasure small worm such as control thrips etc., also is the important component part of the insect comprehensive regulation (IPM).Before using to the field, any medicament delivery must therefore explore the basis that simple and practical effective bioassay method just becomes chemical control through indoor screening process.

The method of medicament biologicall test commonly used comprises drop method, leaf dipping method, dip method, glass medicine embrane method etc. at present, these methods are fit to the effective dose at the bigger insect test medicament of volume, but said method is for the longest only 1 millimeter insect is all inapplicable.Such as in recent years, the harm on vegetables and flowers is year by year the thrips of the trend that rises, calendar year 2001 palm thrips find first in northern China, and protection ground vegetables are caused serious threat.A kind of new invasion insect-Frankliniella occidentalis invasion China in 2003, because its invasion property is strong, harm extensively causes the extensive concern of all circles.Ou Xiaoming 1996 (" measuring the drug-fast a kind of new technology of thrips ". pesticide science and management, 1996 (4): 23-24) proposed a kind of glass residual treatment method, be used to measure the resistance to the action of a drug of thrips, but this method step is many, also need the Potter spray tower as utility appliance, the Potter spray tower because of cost an arm and a leg, complicated operation is difficult for being adopted by most of units.Rueda and Shelton (RuedaA and Shelton A M.Development of a bioassay system for monitoring susceptibilityin in 2003 Thrips tabaci.Pest Manag.Sci.2003,59:553-558.) a kind of bioassay method-TIBS at thrips (thrips insectcide bioassay system) proposed, this method equipment is simple, be convenient to operation, be applicable to biologicall test to field thrips population, by the chemicals treatment internal tank, use vacuum pump to draw insect in container, the medicine film that insect is touched keep away in the container and play action of contace poison, it can carry out evaluating drug effect very accurately to the medicament with contact toxicity, is suitable for the medicament biologicall test of little insect.

Though but this method has solved the difficulty that exists in the medicament biologicall test of small size insect, but all there is a defective: do not consider that a lot of medicaments not only have contact toxicity for insect with above-mentioned additive method, tag for having simultaneously, the medicament of stomach poison function, can not measure the effective active of such medicament truly, the dosage that causes testing out substantially exceeds lethal dose, environmental pollution is serious, also wastes medicament, returns insect and has applied bigger selection pressure.For the bigger insect of volume, there is not unworkable problem, the tester generally can directly be immersed in insect the contact toxicity of testing drug in the medicament, perhaps directly give plant tissue that the agent of insect medicine feed soaked to test the stomach poison activity of medicament, but still do not take into account the situation of tagging and working simultaneously with stomach toxicity, the pharmaceutical quantities that causes giving when the bigger insect of control volume also is much higher than required, pollutes self-evident especially during dispenser to environment.

Summary of the invention

At the defective in the above-mentioned field, the invention provides widely applicable, the new method---called after leaf pipe medicine embrane method that all can effectively measure the medicament of tagging, stomach toxicity acts on simultaneously, this method can accurately determine the minimum effective dose of a kind of medicament for insect, can effectively kill insect, alleviate to top the waste of environmental pollution and medicament again.

The bioassay method of a kind of insect, it is characterized in that be built-in with insect and inwall be attached with the plant tissue of placing the insect happiness food of crossing through the same concentrations chemicals treatment in the container of medicament.

The container that described inwall adheres to medicament is to obtain by medicament soaking container inside.

Described chemicals treatment refers to that medicament soaks plant tissue.

Described built-in insect can move into utensils such as tweezers or writing brushes.

Described container bottom has aperture, and the air-flow that built-in insect can adopt vacuum pump to produce sucks in the container.

Described container is a plastic centrifuge tube.

Described plastic centrifuge tube is the centrifuge tube that seal closure is arranged of 0.5ml or 1.5ML.

Described insect is the thrips class pest.

Described plant is the parasitic plant of insect.

Medicament assay method provided by the invention is measured the minimum effective dose of tested medicament to the combined action mode of insect with medicine, be attached with the plant tissue of placing the insect happiness food of crossing through chemicals treatment in the container of medicament at inwall, after putting into insect, insect makes medicament to its action of contace poison onset by the medicine film of contacting container inwall, the vegetable material of putting into plays the effect of feeding insect on the one hand, makes medicament performance stomach poison function on the other hand by being eaten.For medicament with fumigation action, can adopt closed container, can guarantee action of contace poison, stomach poison function, fumigation action simultaneously effectively, thereby can estimate accurately and effectively by the effect of reagent medicament.

The used container of this method can make its inwall adhere to the medicine film with a lot of modes, is pharmaceutical formulation and deciding, and the present invention adopts medicament soaking container inside to make inwall adhere to one deck medicine film.Also can adopt a lot of modes to make it become the carrier of medicament for the processing of vegetable material, for example spray, soak, use the simplest immersion process among the present invention.

For tested insect, volume is if bigger, big can not cause damage to playing vitality to it being transferred to tweezers or soft writing brush in the container, then directly it is moved in the container with tweezers or writing brush, and for the little insect that is difficult to differentiate one by one to naked eyes of volume, as the thrips class pest, the present invention has adopted the air-flow of vacuum pump insect to be sucked the method for container, this moment, used container was the foraminate pipe in bottom, insect, is sealed aperture with sealing film in aperture is sent into container then by air-flow.Vitality to insect when having avoided in the classic method with the direct picking insect of writing brush or tweezers like this causes damage and influence accurately to measure, and is easy to operate.

The present invention adopts the centrifuge tube of laboratory plastic tape gland bonnet commonly used, and this class pipe use cost is low, be easy to obtain, and the aperture of the convenient fluke of puncturing out in its bottom.

The used container of medicament bioassay method provided by the invention is decided on the volume of tested insect, insect for aphid, the small volume of thrips class, 1.5ML or the plastic centrifuge tube of 0.5ML just can reach requirement, and double dish, test tube or the triangular flask etc. that needn't use glass or plastics are as experiment container, these need take than large space, and many glasswares are damaged easily, scrub inconvenience, the effort of taking a lot of work.

Bioassay method of the present invention is a kind of method efficiently and effectively for control r-countermeasure small worm, particularly this harm of thrips is serious, the insect that is difficult to prevent and treat, therefore as the examination worm, evidence medicament biological test method of the present invention can provide effective medication standard fast for control kind of a class pest with thrips in the present invention.

Used the edible vegetable material of insect in the drug monitoring method provided by the invention, this vegetable material preferably tested insect host plant and be that it likes the tissue that eats, like this can be farthest near the environmental baseline of pest control in the actual production, extremely important for the effect of accurate mensuration medicament.

Embodiment

Embodiment 1 leaf pipe medicine embrane method

Step 1: soak pipe: medicament preparation 6-7 concentration such as table 1, other adds 1 blank, and each concentration liquid is filled with 4 of 1.5mL centrifuge tubes respectively, and every is 1 repetition, with good corresponding contrast of marking pen mark and concentration of treatment, places 4h.Outwell soup then, centrifuge tube is placed on the experiment table dry, stand-by.

Step 2: soak leaf: the fresh cabbage leaves wash clean that will gather, dry.Flooded for 10 seconds in each concentration liquid respectively, the blade after the chemicals treatment is put on the experiment table and dried, and is stand-by.The cabbage leaves that will dry with scissors is cut into little of the rectangle of 2cm * 4cm, sandwiches with pincet in the centrifuge tube of corresponding liquor strength, and 1 of every pipe is placed smooth.

Step 3: fluke: the fine needle needle point that will be about about 10cm heats through spirit lamp, at the bottom of the centrifuge tube pipe that thorn dries, scalds the hole of the about 2-3mm of diameter.The interface of vacuum pump is sealed with gauze, is enclosed within the above centrifuge tube mouth of pipe of handling well, opens vacuum pump, regulates air-flow size (being advisable just can pick up the examination worm), and thrips is aimed in the boiling hot hole at the bottom of the centrifuge tube pipe, makes the worm favorable current suck centrifuge tube.Contrast aftertreatment earlier, every pipe inhale 2 age nymph 10-20 head, build the pipe lid then, seal boiling hot hole with sealing film.Place in 25 ℃ of constant temperature ovens illumination 16L:8D, check result behind 48h at last.

Step 4: check result: insect in the centrifuge tube and blade are poured out on blank sheet of paper contrast aftertreatment earlier.Judge that death standard is, touch polypide with writing brush, can not creep is considered as death.Insect sum under the record respective concentration in every pipe, live borer population and dead borer population.

Test figure is obtained virulence regression equation and LC with reference to probit value Finney analytic approach ₅₀Equivalent.Calculate the mortality ratio of each insect of handling and use the significance of difference between the SAS software analysis different disposal (distinct methods and variable concentrations).Table 1 has compared to be measured 10% with leaf pipe medicine embrane method and TIBS method and eliminates the toxic effect of SC to Frankliniella occidentalis, and the main mode of action that eliminates is stomach poison function.

Table 1 Frankliniella occidentalis is in 10% mortality ratio that eliminates under SC different disposal concentration and the different disposal method

Annotate: lowercase represents that Frankliniella occidentalis is between different disposal concentration; Capitalization represents that Frankliniella occidentalis is between medicament different disposal method

As can be seen, the mortality ratio that adopts leaf pipe medicine embrane method to handle Frankliniella occidentalis improves with the raising of handling concentration, and indifference between the TIBS method variable concentrations, and do not have regularity, can not truly reflect the actual insecticidal activity that eliminates.

The checking of embodiment 2 leaf pipe medicine embrane methods

Test medicine:

Organic phosphates: 40% chlopyrifos EC (Shandong Dacheng Farm Chemical Co., Ltd.), pyrethroid: 4.5% effective cypermethrin EC (Shanghai positive magnificent agricultural chemicals company limited), 20% kills chrysanthemum ester EC (Jiangsu grain sun insecticide factory), microbiology class pesticide: 1.8% avermectin (Avermectin) EC (Beijing ZhongNon Da Biology Technology Co., Ltd), 2.5% dish happiness (pleocidin) SC (The Dow Agrosciences, LLC.) emamectin-benzoate ME, botanical pesticide: 0.3% matrine AS (No. 58, the full true road of spring industry park collection, Yinchuan City), novel pesticide: 35% Imidacloprid SC (Jiangsu Changlong Chemical Co), 10% Imidacloprid WP (Suzhou Hua Yuan Group Co.,Ltd), 25% Acker Thailand (thiophene worm piperazine) WDGS (just reaching earlier (China) Investment Co., Ltd), 10% eliminates (chlorfenapyr) SC (the cyanamide world), 5% sharp strength spy (fluorine worm nitrile) SC (Bayer Cropscience Co.,Ltd).

The result is as shown in table 2,40% chlopyrifos EC, 4.5% effective cypermethrin EC, 20% kill chrysanthemum ester EC, 1.8% avermectin (Avermectin) EC, the happiness of 2.5% dish (pleocidin) SC, 3% emamectin-benzoate ME after 24 hours, 0.3% matrine AS, 10% eliminates (chlorfenapyr) SC, 5% sharp strength spy (fluorine worm nitrile) SC virulence effect is better, LC ₅₀Be respectively 7.0,13.0,13.0,2.0,0.2,0.5,3.5,25.6 and 1.6mg/L, can be used as control Frankliniella occidentalis effective agent.The contrast mortality ratio of various medicaments is all at 0-6.9%, and the contrast mortality ratio that meets the biologicall test requirement is controlled at 10% with interior requirement, further illustrates the reliability and the validity of this method.

The different medicaments of table 2 are to the indoor biometrics result of Frankliniella occidentalis

Claims

1, the bioassay method of a kind of insect, it is characterized in that be built-in with insect and inwall be attached with the plant tissue of placing the insect happiness food of crossing through the same concentrations chemicals treatment in the container of medicament.

2, the container that the bioassay method of insect according to claim 1, described inwall adhere to medicament is to obtain by medicament soaking container inside.

3, the bioassay method of insect according to claim 1, described chemicals treatment refer to that medicament soaks plant tissue.

4, the bioassay method of insect according to claim 1, described built-in insect can move into utensils such as tweezers or writing brushes.

5, the bioassay method of insect according to claim 1, described container bottom has aperture, and the air-flow that built-in insect can adopt vacuum pump to produce sucks in the container.

6, the bioassay method of insect according to claim 5, described container are plastic centrifuge tube.

7, the bioassay method of insect according to claim 6, described plastic centrifuge tube are the centrifuge tube that seal closure is arranged of 0.5ml or 1.5ML.

8, the bioassay method of insect according to claim 5, described insect are the thrips class pest.

9, the bioassay method of insect according to claim 1, described plant are the parasitic plant of insect.