CN104472443A - Indoor biological activity assay method of orange ceroplastes rubens maskell and application of indoor biological activity assay method - Google Patents
Indoor biological activity assay method of orange ceroplastes rubens maskell and application of indoor biological activity assay method Download PDFInfo
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- CN104472443A CN104472443A CN201410842535.3A CN201410842535A CN104472443A CN 104472443 A CN104472443 A CN 104472443A CN 201410842535 A CN201410842535 A CN 201410842535A CN 104472443 A CN104472443 A CN 104472443A
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- ceroplastes rubens
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- 241001450756 Ceroplastes rubens Species 0.000 title claims abstract description 43
- 230000004071 biological effect Effects 0.000 title claims abstract description 15
- 238000003556 assay Methods 0.000 title abstract 3
- 238000000034 method Methods 0.000 claims abstract description 27
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 7
- 241000207199 Citrus Species 0.000 claims description 33
- 235000020971 citrus fruits Nutrition 0.000 claims description 33
- 241000196324 Embryophyta Species 0.000 claims description 32
- 239000003814 drug Substances 0.000 claims description 17
- 239000007788 liquid Substances 0.000 claims description 10
- 230000006378 damage Effects 0.000 claims description 8
- 238000011081 inoculation Methods 0.000 claims description 6
- 208000027418 Wounds and injury Diseases 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 208000014674 injury Diseases 0.000 claims description 4
- 238000007689 inspection Methods 0.000 claims description 3
- 238000012360 testing method Methods 0.000 abstract description 17
- 241000238631 Hexapoda Species 0.000 abstract description 9
- 238000004166 bioassay Methods 0.000 abstract description 6
- 235000013305 food Nutrition 0.000 abstract description 4
- 230000012010 growth Effects 0.000 abstract description 3
- 235000015097 nutrients Nutrition 0.000 abstract description 3
- 230000004083 survival effect Effects 0.000 abstract description 3
- 239000000575 pesticide Substances 0.000 abstract 5
- 238000007789 sealing Methods 0.000 abstract 1
- 231100000225 lethality Toxicity 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 239000003905 agrochemical Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- NHDHVHZZCFYRSB-UHFFFAOYSA-N pyriproxyfen Chemical compound C=1C=CC=NC=1OC(C)COC(C=C1)=CC=C1OC1=CC=CC=C1 NHDHVHZZCFYRSB-UHFFFAOYSA-N 0.000 description 3
- WCXDHFDTOYPNIE-RIYZIHGNSA-N (E)-acetamiprid Chemical compound N#C/N=C(\C)N(C)CC1=CC=C(Cl)N=C1 WCXDHFDTOYPNIE-RIYZIHGNSA-N 0.000 description 2
- 239000005875 Acetamiprid Substances 0.000 description 2
- 241001124076 Aphididae Species 0.000 description 2
- 239000005658 Tebufenpyrad Substances 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 239000004530 micro-emulsion Substances 0.000 description 2
- 239000012452 mother liquor Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000004088 simulation Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- ZZYSLNWGKKDOML-UHFFFAOYSA-N tebufenpyrad Chemical compound CCC1=NN(C)C(C(=O)NCC=2C=CC(=CC=2)C(C)(C)C)=C1Cl ZZYSLNWGKKDOML-UHFFFAOYSA-N 0.000 description 2
- 230000001018 virulence Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000178220 Anicetus beneficus Species 0.000 description 1
- 241000178219 Anicetus ohgushii Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000317929 Chalcididae Species 0.000 description 1
- 241000675108 Citrus tangerina Species 0.000 description 1
- 235000015001 Cucumis melo var inodorus Nutrition 0.000 description 1
- 240000002495 Cucumis melo var. inodorus Species 0.000 description 1
- 239000005947 Dimethoate Substances 0.000 description 1
- 241001147399 Encyrtidae Species 0.000 description 1
- 241001468872 Fumagospora capnodioides Species 0.000 description 1
- 206010021703 Indifference Diseases 0.000 description 1
- 241000270322 Lepidosauria Species 0.000 description 1
- 241000819999 Nymphes Species 0.000 description 1
- 239000005663 Pyridaben Substances 0.000 description 1
- 239000005927 Pyriproxyfen Substances 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000000205 computational method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- MCWXGJITAZMZEV-UHFFFAOYSA-N dimethoate Chemical compound CNC(=O)CSP(=S)(OC)OC MCWXGJITAZMZEV-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 231100000636 lethal dose Toxicity 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- DWFZBUWUXWZWKD-UHFFFAOYSA-N pyridaben Chemical compound C1=CC(C(C)(C)C)=CC=C1CSC1=C(Cl)C(=O)N(C(C)(C)C)N=C1 DWFZBUWUXWZWKD-UHFFFAOYSA-N 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Ecology (AREA)
- Forests & Forestry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Health & Medical Sciences (AREA)
- Toxicology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Botany (AREA)
- Cultivation Of Plants (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses an indoor biological activity assay method of orange ceroplastes rubens maskell. The method comprises the following steps of a step of naturally inoculating and feeding test insects; a step of preparing a test reagent into a pesticide solution with the predetermined concentration; a step of immersing branches with insects of potted seedlings in the prepared pesticide solution, then sealing the immersed branches with the insects by using a thin and soft gauze screen; a step of placing the potted seedlings immersed with a pesticide solution in an artificial climate chamber in a light-dark period with a ratio of light to dark (L to D) of 14 to 10h, temperature of 25+/-1 DEG C and relative humidity of 75%; a step of checking death and survival of tested ceroplastes rubens maskells after feeding for 24 hours, and recording the death number and the survival number. According to the method, the test insects immersed in the pesticide solution are cultured by using living plants; the freshness of food of the orange ceroplastes rubens maskells is ensured; sufficient nutrient is provided for the orange ceroplastes rubens maskells; the test is carried out by simulating the natural growth environment condition of the orange ceroplastes rubens maskells; the accuracy and the precision of the bioassay results can be effectively improved; the resistance level of the orange ceroplastes rubens maskell group can be comprehensively, accurately and precisely known; reference is accurately provided for reasonably using the pesticides and making resistance treating scheme.
Description
Technical field
The present invention relates to insect pest of the plant prevention and control field, particularly a kind of citrus ceroplastes rubens indoor biological activity determination method and application thereof.
Background technology
Ceroplastes rubens harm plant not only branch when being injured serious, blade is covered with polypide, draw juice in a large number, make plant growth gesture weak, and secrete honeydew in this worm growth and development process, a large amount of tillers bringing out sooty mould bacterium are raw, branches and leaves surface forms the mould layer of black, both make plant be subject to the dual harm of disease worm, affect again plant photosynthesis and respiration thus to accelerate plant weak, even withered.Have a strong impact on greening and ornamental value.Egg hatch contains the phase in mid-June, just incubates that nymph is many to climb from parent at fine day noon, if overcast and rainy meeting is about parent shell creeps half an hour, after anchor at successively on branches and leaves and endanger.The parasite of ceroplastes rubens is more, common are Anicetus beneficus, Anicetus ohgushii Tachikawa, the flat angle of lecanium (short-tail) encyrtid, relies the soft a red-spotted lizard aphid chalcid fly of food etc.Indoor bioassay be a kind of can more comprehensively, more accurately understand the means of insect field population resistance level, also be a kind of method of comparatively traditional understanding resistance, now widely used various bioassay method, seedling method, drop method is soaked as being with leaf, to large for examination insect damage, contrast lethality is large, the raw accuracy surveying result of impact.
Summary of the invention
In view of this, the object of the present invention is to provide a kind of citrus ceroplastes rubens indoor biological activity determination method and application thereof, live plant is adopted to cultivate the examination worm soaking liquid, ensure the fresh of citrus ceroplastes rubens food and provide sufficient nutrient for citrus ceroplastes rubens, test under the natural environment growth conditions residing for citrus ceroplastes rubens, the accuracy of effective raising bioassay results and accuracy, thus comparatively comprehensively accurate and accurate understanding citrus ceroplastes rubens population resistance level, for reasonable employment agricultural chemicals, formulate resistance management scheme and provide foundation comparatively accurately.
Citrus ceroplastes rubens indoor biological activity determination method of the present invention, comprises the following steps:
A. try worm and naturally inoculate raising: contain aspire to the serious band worm orange branches of field acquisition occurrence injury at citrus ceroplastes rubens egg hatch, band worm branch is docked on potted plant nursery stock to be tested;
B. make up a prescription: liquid reagent agent being mixed with setting concentration;
C. medicine is soaked: be soaked in the liquid prepared by whole for the band worm branch of potted plant nursery stock;
D. indoor feeding: the potted plant nursery stock of dipped medicament is placed on light dark period L:D=14: 10h, temperature is 25 ± 1 DEG C, and relative moisture is raise in the phytotron of 75%;
E. result inspection: check the death of tested citrus ceroplastes rubens after raising 24h and survive situation, records its death toll and survives number;
Further, in step a, by band worm branch by colligation on the branch of potted plant nursery stock, then potted plant nursery stock is moved into airtight solarium and carries out nature inoculation and raise;
Further, in step c, the soak time of described band worm branch is 10 seconds.
The present invention also discloses the application of a kind of citrus ceroplastes rubens indoor biological activity determination method in plant protection art.
Beneficial effect of the present invention: a kind of citrus ceroplastes rubens indoor biological activity determination method of the present invention and application thereof, band worm branch is utilized to carry out nature inoculation by bind in the mode of nursery stock branch to be tested, survival rate is higher, and do not damage examination worm, live plant is adopted to cultivate the examination worm soaking liquid, ensure the fresh of citrus ceroplastes rubens food and provide sufficient nutrient for citrus ceroplastes rubens, by carrying out under self-sow environmental condition residing for simulation citrus ceroplastes rubens testing by testing under self-sow environmental condition residing for simulation citrus ceroplastes rubens, the accuracy of effective raising bioassay results and accuracy, thus comparatively comprehensively accurate and accurate understanding citrus ceroplastes rubens population resistance level, for reasonable employment agricultural chemicals, formulate resistance management scheme and provide foundation comparatively accurately.
Embodiment
The citrus ceroplastes rubens indoor biological activity determination method of the present embodiment, comprises the following steps:
A. try worm and naturally inoculate raising: contain aspire to the serious band worm orange branches of field acquisition occurrence injury at citrus ceroplastes rubens egg hatch, band worm branch is docked on potted plant nursery stock to be tested;
B. make up a prescription: liquid reagent agent being mixed with setting concentration;
C. medicine is soaked: be soaked in the liquid prepared by whole for the band worm branch of potted plant nursery stock;
D. indoor feeding: the potted plant nursery stock of dipped medicament is placed on light dark period L:D=14: 10h, temperature is 25 ± 1 DEG C, and relative moisture is raise in the phytotron of 75%;
E. result inspection: check the death of tested citrus ceroplastes rubens after raising 24h and survive situation, records its death toll and survives number; Adopt the band worm branch and branch live plant leaching medicine the mode of raising that soak potted plant nursery stock, the self-sow environment of citrus ceroplastes rubens is simulated with this, that gas permeability and institute's phytophagous freshness etc. are all with natural conditions and indifference, accuracy and the accuracy of bioassay results is improved with this, thus comparatively comprehensively accurate and accurate understanding citrus ceroplastes rubens population resistance level, for reasonable employment agricultural chemicals, formulation resistance management scheme provide foundation comparatively accurately.
In the present embodiment, the soak time of described band worm branch is 10 seconds;
In the present embodiment, in step a, by band worm branch by colligation on the branch of potted plant nursery stock, then potted plant nursery stock is moved into airtight solarium and carries out nature inoculation and raise.
This citrus ceroplastes rubens indoor biological activity determination method is applied in plant protection art.
Below by specific embodiment, the present invention is further elaborated.
Embodiment one
1. try worm condition of culture
The phase is contained in mid-June (Chongqing June) at citrus ceroplastes rubens egg hatch, the orange branches that field acquisition occurrence injury is serious, take back indoor, the branch of clip appropriate length, tie up and naturally inoculate on indoor pot nursery stock, then concentrate a large airtight solarium to carry out nature inoculation potted plant nursery stock to raise preparation and test, test after insect transfer inoculation, during test, the ceroplastes rubens selecting enough tests from generation to generation neat potted plant nursery stock branch occurs and blade is tested, reject inactive and generation irregular individuality, only retain and grow more neat individuality from generation to generation, then be dipped in the liquid of the variable concentrations prepared whole for band worm branch, dip time 10 seconds, the process of each medicament variable concentrations cross nursery stock concentrate be stored in artificial climate room, be convenient to the death toll of medicine " Invest, Then Investigate " aphid, then the potted plant nursery stock of dipped medicament being placed on light dark period is 14: 10h, (25 ± 1) DEG C, in the phytotron of relative moisture 75%, often process and repeat for 4 times, lethality is checked after normal raising 24h, laboratory is gone back to the branch of the dipped medicament of scissors clip, lethality is checked with pin.
2. reagent
2.1 test medicine
10% Nylar microemulsion
Common name: pyriproxyfen is provided by client
2.2 contrast medicaments
10% Acetamiprid ether microemulsion
Common name dimethoate is provided by client
2.3 other reagent
3 test process
3.1 dosage are arranged
3.2 tests repeat
Test arranges 4 repetitions, the insect population of each repetition more than 30.
3.3 processing mode
3.3.1 processing time and number of times
Adopt infusion process to process the citrus ceroplastes rubens of raising, 24 hours " Invest, Then Investigate " death condition, carry out statistical analysis.
3.3.2 apparatus and administrated method is used
With graduated cylinder, first the apparatuses such as beaker will be diluted to the mother liquor of desired concn for the former medicine of examination, then dilute with water mother liquor is to the concentration preset, and application infusion process processes examination worm.
3.3.2 processing method
Two medicaments are made into 5 ~ 7 diluted concentrations according to a certain percentage respectively, the best dilution intervals that reagent agent test is selected in preliminary experiment is first done before test, branch with citrus ceroplastes rubens immerses in the liquid converted and soaks 10 seconds by the concentration range selected according to preliminary experiment, separately do 4 leaching clear water in contrast, then dipped potted plant nursery stock being placed on light dark period is 14: 10h, (25 ± 1) DEG C, in the phytotron of relative moisture 75%, often process and repeat for 4 times, lethality is checked after normal raising 24h, laboratory is gone back to the net branch of the dipped medicament of scissors clip, lethality is checked with pin.Virulence regression equation, LC is calculated with probit analysis
50, standard error is equivalent.
4. computational methods
Become by concentration conversion ppm to input calculator (the irm software that plant protection institute of Agricultural University Of Southwest makes) later, obtain virulence regression equation, LC
50value, standard error SD, according to LC
50value and standard error calculate 95% confidence limit (see table 1).
5. evaluate
The LC of 10% Nylar
50value is greater than contrast medicament 10% Acetamiprid ether, illustrate that its activity is lower, and its b value is little compared with pyridaben, and illustrate that degree of scatter is comparatively large, citrus Jie-Li enzyme-SQ is larger to its response difference.
Table 1 tebufenpyrad is to tangerine full pawl ceroplastes rubens indoor biological determination of activity regression equation, the lethal concentration of 50 and standard error
2013
Table 2 tebufenpyrad is to Jie-Li enzyme-SQ indoor biological determination of activity raw data table
2013
What finally illustrate is, above embodiment is only in order to illustrate technical scheme of the present invention and unrestricted, although with reference to preferred embodiment to invention has been detailed description, those of ordinary skill in the art is to be understood that, can modify to technical scheme of the present invention or equivalent replacement, and not departing from aim and the scope of technical solution of the present invention, it all should be encompassed in the middle of right of the present invention.
Claims (4)
1. a citrus ceroplastes rubens indoor biological activity determination method, is characterized in that: comprise the following steps:
A. try worm and naturally inoculate raising: contain aspire to the serious band worm orange branches of field acquisition occurrence injury at citrus ceroplastes rubens egg hatch, band worm branch is docked on potted plant nursery stock to be tested;
B. make up a prescription: liquid reagent agent being mixed with setting concentration;
C. medicine is soaked: be soaked in the liquid prepared by whole for the band worm branch of potted plant nursery stock;
D. indoor feeding: the potted plant nursery stock of dipped medicament is placed on light dark period L:D=14: 10h, temperature is 25 ± 1 DEG C, and relative moisture is raise in the phytotron of 75%;
E. result inspection: check the death of tested citrus ceroplastes rubens after raising 24h and survive situation, records its death toll and survives number.
2. citrus ceroplastes rubens indoor biological activity determination method according to claim 1, is characterized in that: in step a, by band worm branch by colligation on the branch of potted plant nursery stock, then potted plant nursery stock is moved into airtight solarium and carries out nature inoculation and raise.
3. citrus ceroplastes rubens indoor biological activity determination method according to claim 2, is characterized in that: in step c, and the soak time of described band worm branch is 10 seconds.
4. the application of citrus ceroplastes rubens indoor biological activity determination method in plant protection art described in any one of claims 1 to 3.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113068526A (en) * | 2021-04-01 | 2021-07-06 | 保山市林业和草原技术推广站(保山市林业和草原科学研究所) | Observation and research method for migratory life history of scale insects |
| CN114600840A (en) * | 2022-02-25 | 2022-06-10 | 广西特色作物研究院 | Method for determining biological activity of diaphorina citri nymphs |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114600840A (en) * | 2022-02-25 | 2022-06-10 | 广西特色作物研究院 | Method for determining biological activity of diaphorina citri nymphs |
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