CN110024921A - A kind of application of biological stain dimethylbenzene aniline in the feeding of thrips class insect film - Google Patents
A kind of application of biological stain dimethylbenzene aniline in the feeding of thrips class insect film Download PDFInfo
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- CN110024921A CN110024921A CN201910339402.7A CN201910339402A CN110024921A CN 110024921 A CN110024921 A CN 110024921A CN 201910339402 A CN201910339402 A CN 201910339402A CN 110024921 A CN110024921 A CN 110024921A
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- thrips
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/179—Colouring agents, e.g. pigmenting or dyeing agents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/90—Feeding-stuffs specially adapted for particular animals for insects, e.g. bees or silkworms
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Food Science & Technology (AREA)
- Insects & Arthropods (AREA)
- Birds (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
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Abstract
The present invention provides a kind of application of biological stain dimethylbenzene aniline in the feeding of thrips class insect film.It comprises the concrete steps that the phosphate buffer that dimethylbenzene aniline biological stain powder is dissolved in the 0.01mol/L of pH value 7.0 ~ 7.4, obtains 1 ~ 10mg/ml biological stain solution, film feeding is carried out to obtain the thrips nymph of dyeing to the thrips nymph of single tube raising.The indicator when present invention is fed using dimethylbenzene aniline as thrips class insect film, biological body protein can non-specifically be combined, positive evidence can be provided for accurate judgement thrips class insect's food-taking under safe concentration, rapid and handy establishes thrips class pest RAN perturbation technique system.
Description
Technical field
The invention belongs to insect organism colouring arts, and in particular to a kind of biological stain dimethylbenzene aniline is in thrips class
Application in the feeding of insect film.
Background technique
Palm thrips (Thrips palmi Karny) belong to Thysanoptera (Thysanoptera) Thripidae (Thripidae)
Thrips (Thrips).Palm thrips are a kind of dangerous worldwide customs quarantine control pests of tool, can endanger more than 600 kinds of plants
Object, it is even more serious to a variety of important agricultural economy crop harm.Last century the seventies palm thrips are for the first time in the platform in China
Gulf area is found, hereafter in the successive occurrence injury in the area such as Guangdong, Hunan, Sichuan, Yunnan, Tibet, Shandong, Beijing.
Research currently for thrips raising aspect is less, is only limitted to gene cloning, protein induced expression and gene space-time
The basic research such as distribution, and the research of film feeding is had not been reported, present invention application dimethylbenzene aniline is dyed as thrips film
The research of indicator also belongs to pioneering.Dimethylbenzene aniline is as a kind of biological stain, alias dimethylbenzene blueness FF；Xylene blue,
Molecular formula: C25H27N2NaO6S2, appearance (character) is black blue powders, shown in purity about 75%(Fig. 1).Dimethylbenzene aniline is in fish
Belong to the minimum substance of toxicity in the acute toxicity test of class, can be used as oxidation-reduction indicator (orange-yellow~yellow), nucleic acid
Electrophoresis indicator or biological stain, but need to carry out accurate control in usage amount, make in order to avoid occurring to poison to insect
With influencing scientific research.
The advantage of present invention combination dimethylbenzene aniline biological dye -- biological body protein can be non-specifically combined, will be given birth to
Influence control of the object dyestuff to insect vital signs is in safe range.Inventor has comprehensively considered dimethylbenzene aniline and has been applied to refer to
The effect for showing the feeding of thrips film, the low rate of feeding during thrips film is fed that successfully solves lead to the problem of feeding effect difference,
The development in the fields such as Fluorometric assay of fluorescence-labeled technology and life science of promotion.
Summary of the invention
The present invention has well solved application of the biological stain dimethylbenzene aniline in the feeding of thrips class insect film, a side
Face can provide technological guidance for thrips class pest dye marker, on the other hand also can detecte film feeding method in thrips class etc.
The assessment of rasping-sucking mouthparts insect's food-taking effect.
To achieve the above object, the present invention adopts the following technical scheme:
A kind of application of biological stain dimethylbenzene aniline in the feeding of thrips class insect film, comprising the following steps:
(1) dimethylbenzene aniline biological stain powder is dissolved in the phosphate buffer of 0.01mol/L, obtains the life of various concentration
Object staining solution；
(2) biological stain solution in the step (1) is subjected to film feeding to thrips nymph, at 25 ± 2 DEG C, humidity 75%, light
According to 14L:10D, under the conditions of feed the thrips nymph for 24 hours, dyed；
(3) staining conditions of label thrips nymph in the step (2) are subjected to imaging of taking pictures with microscope.
The pH value of the 0.01mol/L phosphate buffer is 7.0~7.4.
The worm age range of the thrips nymph is 1 age ~ 2 ages.
The concentration of the biological stain solution is 1-10mg/ml.
According to the acquisition methods of thrips in the step (2), comprising the following steps: after thrips nymph Nature enemy 2h,
It is chosen with writing brush into the raising pipe of open double-faced, both ends all use Parafilm film to seal, and wherein configuration good colourant will be added dropwise in one end
Moisturizing is covered with Parafilm film again after solution.
The present invention has the advantages that
The invention discloses a kind of application of biological stain dimethylbenzene aniline in the feeding of thrips class insect film, by dimethylphenyl phenyl
Biological stain solution is obtained after the mixing of the phosphate buffer of amine biological stain powder and 0.01mol/L, to the Ji of single tube raising
Horse nymph carries out film feeding to obtain the thrips nymph of dyeing.In the case where guaranteeing safe, harmless to thrips, pass through dyeing
The acid-base property two indices of agent concentration and buffer go evaluation dimethylbenzene aniline as the effect of thrips class insect film feeding indicator
Fruit successfully realizes the biological dye indicator for the first time and monitors feasibility to the feeding of the Thysanopteras insect such as thrips class, it can
Non-specifically to combine biological body protein, positive evidence can be provided for accurate judgement thrips class insect's food-taking, rapid and handy
Establish thrips class pest RAN perturbation technique system.
Detailed description of the invention
Fig. 1 dimethylphenyl phenyl amine chemical structural formula.
Fig. 2 is staining conditions of the various concentration biological stain liquid to thrips.
Fig. 3 is influence of the dimethylbenzene aniline feeding to thrips survival rate.
Fig. 4 is to add biological stain and water respectively to carry out film feeding to thrips.
Fig. 5 is that single tube raises thrips, is fritter kidney bean in pipe.
Specific embodiment
The present invention is further illustrated by the following examples, but protection scope of the present invention is not limited to following reality
Apply example.Dimethylbenzene aniline used in the present embodiment is known compound, can be obtained by commercial sources.
Embodiment 1
(1) dimethylbenzene aniline biological stain powder is dissolved in the phosphate buffer of 0.01mol/L, the biology dye of 100mg/ml
Toner solution；
(2) phosphate buffer for the 0.01mol/L for being 7.0 by the dimethylbenzene aniline biological stain and pH value of 100mg/ml presses body
Product is mixed than 1:9, obtains the biological stain solution that concentration is 10mg/ml；
(3) biological stain solution in step (1) is subjected to film feeding to thrips nymph, in 25 DEG C, humidity 75%, illumination 14L:
The thrips nymph dyed for 24 hours is fed under the conditions of 10D；
(4) staining conditions of label thrips nymph in the step (2) are subjected to imaging of taking pictures with microscope.
Embodiment 2
(1) phosphate buffer for the 0.01mol/L for being 6.5 by the dimethylbenzene aniline biological stain and pH value of 100mg/ml presses body
Product is mixed than 1:9, obtains the biological stain solution that concentration is 10mg/ml；
(2) biological stain solution in step (1) is subjected to film feeding to thrips nymph, at 28 DEG C, humidity 75%, illumination 14L:
10D, under the conditions of feed the thrips nymph for 24 hours, dyed；
(3) staining conditions of label thrips nymph in the step (2) are subjected to imaging of taking pictures with microscope.
Embodiment 3
(1) phosphate buffer for the 0.01mol/L for being 7.8 by the dimethylbenzene aniline biological stain and pH value of 100mg/ml presses body
Product is mixed than 1:9, obtains the biological stain solution that concentration is 10mg/ml；
(2) biological stain solution in step (1) is subjected to film feeding to thrips nymph, in 28 DEG C, humidity 75%, illumination 14L:
The thrips nymph dyed for 24 hours is fed under the conditions of 10D；
(3) staining conditions of label thrips nymph in the step (2) are subjected to imaging of taking pictures with microscope.
Embodiment 4
(1) phosphate buffer for the 0.01mol/L for being 7.2 by the dimethylbenzene aniline biological stain and pH value of 10mg/ml presses body
Product is mixed than 1:9, obtains the biological stain solution that concentration is 1mg/ml；
(2) biological stain solution in step (1) is subjected to film feeding to thrips nymph, in 26 DEG C, humidity 75%, illumination 14L:
The thrips nymph dyed for 24 hours is fed under the conditions of 10D；
(3) staining conditions of label thrips nymph in the step (2) are subjected to imaging of taking pictures with microscope.
Embodiment 5
(1) phosphate buffer for the 0.01mol/L for being 7.0 by the dimethylbenzene aniline biological stain and pH value of 100mg/ml presses body
Product is mixed than 1:5, obtains the biological stain solution that concentration is 16mg/ml；
(2) biological stain solution in step (1) is subjected to film feeding to thrips nymph, in 25 DEG C, humidity 75%, illumination 14L:
The thrips nymph dyed for 24 hours is fed under the conditions of 10D；
(3) staining conditions of label thrips nymph in the step (2) are subjected to imaging of taking pictures with microscope.
Comparative example
(1) phosphate buffer for the 0.01mol/L for being directly 7.0 with pH value is feeding raw material, is not added with biological stain diformazan
Benzene aniline obtains the biological stain solution that concentration is 0mg/ml；
(2) by biological stain solution in step (1) to thrips nymph carry out film feeding, 25 DEG C, humidity 75%, under the conditions of feed
For 24 hours, the thrips nymph dyed；
(3) staining conditions of label thrips nymph in the step (2) are subjected to imaging of taking pictures with microscope.
From figure 2 it can be seen that biological stain dimethylbenzene aniline is under various concentration of the present invention compared with compareing thrips
(1mg/mL ~ 10mg/mL) is preferable to the dyeing effect of thrips, be readily viewed from morphology to.Biological stain diformazan
Benzene aniline is not in concentration of the present invention and parameter (10mg/mL, phosphate buffer pH value are 6.5,28 DEG C) and (10mg/mL, phosphoric acid are slow
Fliud flushing pH value is 7.8,28 DEG C) under thrips morphological differences it is larger, thrips is slow in action, and dead, and dimethylbenzene aniline occurs in part
In the case where 16mg/mL phosphate buffer pH value is 7.0, thrips is slow in action, most of dead.When stain concentration is low
Dyeing effect is poor when 1mg/ml, is unfavorable for Germicidal efficacy and monitoring.
In conclusion biological stain reagent of the invention -- dimethylbenzene aniline survival rate in the feeding of thrips film with control group
Close, illustrate that the present invention feeds thrips film influences without generating conspicuousness, can play auxiliary thrips film feeding effect
Effect.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (6)
1. a kind of application of biological stain dimethylbenzene aniline in the feeding of thrips class insect film.
2. application according to claim 1, which comprises the following steps:
(1) dimethylbenzene aniline biological stain powder is dissolved in the phosphate buffer of 0.01mol/L, obtains the diformazan of various concentration
Benzene aniline biological stain solution；
(2) biological stain solution in the step (1) is subjected to film feeding to thrips nymph, at 25 ± 2 DEG C, humidity 75%, light
According to feeding the thrips nymph dyed for 24 hours under conditions of 14L:10D；
(3) staining conditions of label thrips nymph in the step (2) are subjected to imaging of taking pictures with microscope.
3. application according to claim 2, which is characterized in that the pH value of the 0.01mol/L phosphate buffer be 7.0~
7.4。
4. application according to claim 2, which is characterized in that the worm age range of the thrips nymph is 1 age ~ 2 ages.
5. application according to claim 2, which is characterized in that the concentration of the biological stain solution is 1-10mg/ml.
6. application according to claim 2, which is characterized in that according to the acquisition methods of thrips in the step (2) are as follows: will
It after thrips nymph Nature enemy 2h, is chosen with writing brush into the raising pipe of open double-faced, both ends all use Parafilm film to seal, by it
Middle one end covers moisturizing with Parafilm film again after configuration good colourant solution is added dropwise.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910339402.7A CN110024921B (en) | 2019-04-25 | 2019-04-25 | Application of biological stain xylene aniline in thrips insect membrane feeding |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910339402.7A CN110024921B (en) | 2019-04-25 | 2019-04-25 | Application of biological stain xylene aniline in thrips insect membrane feeding |
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| Publication Number | Publication Date |
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| CN110024921A true CN110024921A (en) | 2019-07-19 |
| CN110024921B CN110024921B (en) | 2022-10-14 |
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| CN201910339402.7A Active CN110024921B (en) | 2019-04-25 | 2019-04-25 | Application of biological stain xylene aniline in thrips insect membrane feeding |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102524188A (en) * | 2012-01-20 | 2012-07-04 | 福建省农业科学院植物保护研究所 | Insect breeding bottom and breeding method of gynaikothrips ficorum |
| CN102640729A (en) * | 2012-04-06 | 2012-08-22 | 山东省农业科学院植物保护研究所 | Method for breeding frankliniella occidentalis |
| CN104012472A (en) * | 2014-03-12 | 2014-09-03 | 云南农业大学 | Indoor breeding method of thrips |
| CN106975084A (en) * | 2017-04-07 | 2017-07-25 | 中国农业科学院植物保护研究所 | A kind of colouring method of tetranychid digestive system |
-
2019
- 2019-04-25 CN CN201910339402.7A patent/CN110024921B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102524188A (en) * | 2012-01-20 | 2012-07-04 | 福建省农业科学院植物保护研究所 | Insect breeding bottom and breeding method of gynaikothrips ficorum |
| CN102640729A (en) * | 2012-04-06 | 2012-08-22 | 山东省农业科学院植物保护研究所 | Method for breeding frankliniella occidentalis |
| CN104012472A (en) * | 2014-03-12 | 2014-09-03 | 云南农业大学 | Indoor breeding method of thrips |
| CN106975084A (en) * | 2017-04-07 | 2017-07-25 | 中国农业科学院植物保护研究所 | A kind of colouring method of tetranychid digestive system |
Non-Patent Citations (2)
| Title |
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| 中国医药集团上海化学试剂公司: "《试剂手册》", 30 November 2002, 上海科学技术出版社 * |
| 马亚斌 等: "高温对西花蓟马卵巢发育及卵黄蛋白含量的影响", 《昆虫学报》 * |
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