CN102323201A - Staining detection method of eggs of blind stinkbug - Google Patents
Staining detection method of eggs of blind stinkbug Download PDFInfo
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- CN102323201A CN102323201A CN201110144283A CN201110144283A CN102323201A CN 102323201 A CN102323201 A CN 102323201A CN 201110144283 A CN201110144283 A CN 201110144283A CN 201110144283 A CN201110144283 A CN 201110144283A CN 102323201 A CN102323201 A CN 102323201A
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Abstract
The invention provides a staining detection method of eggs of blind stinkbugs, which comprises the following steps of: after impregnating plant materials in eosin solution with the weight in volume concentration of 0.5 to 2 w/v% for 1 to 5 minutes, taking out the plant materials and flushing the planting materials by water; then adsorbing up surface moisture; and observing through naked eyes or carrying out microscopic examination to find that ovum operculum parts of the eggs of the blind stinkbugs are stained into red and have obvious color contrast with the plant materials, so that the number of the eggs of the blind stinkbugs is rapidly and accurately detected. The method is simple, convenient and practical. The problem that in the plant tissues, the eggs of the blind stinkbugs are difficult to survey is solved. The method is suitable to be popularized and applied.
Description
Technical field
The invention belongs to the plant protection technology field, specifically, relate to a kind of dyeing detection method of plant bug ovum.
Background technology
1997, China began commercialization plantation Bt cotton, had reached 3,800,000 hectares to cultivated area in 2008, accounted for 70% of the Cotton in China total area.The harm that the establishing in large scale of Bt cotton has effectively been controlled bollworm Helicoverpa armigera takes place, and has reduced the pesticide dosage of control bollworm.Yet the minimizing of chemical pesticide is used to cotton field plant bug population growth the space is provided, and plant bug has risen into the primary pest on China Bt cotton, and has involved various crop such as jujube tree.China's plant bug kind is numerous, and dominant species comprises green plant bug Apolygus lucorum (Meyer-D ü r.), black striped plant bug Adelphocoris suturalis (Jakovlev), adelphocoris taeniophorus Adelphocoris fasiaticollis (Reuter), alfalfa plant bug Adelphocoris lineolatus (Goeze) and tarnished plant bug Lygus pratensis (L.).Plant bug lays eggs among plant tissue, only stays ovum operculum at plant surface, the long not enough 1cm of ovum operculum, and lighter color, even therefore under Stereo microscope, also quickly and accurately it is observed, counts than difficulty, this makes the investigation work of plant bug ovum amount be difficult to carry out.
Summary of the invention
The dyeing detection method that the purpose of this invention is to provide a kind of ovum of plant bug fast and accurately.
In order to realize the object of the invention, the dyeing detection method of a kind of plant bug ovum of the present invention, it is that vegetable material impregnated in the 0.5-2w/v% eosin solution after 1-5 minute, takes out the water flushing, blots surface moisture then, visual inspection or microscopy counting.Preferably, it is that vegetable material impregnated in the 1w/v% eosin solution 2 minutes.What wherein, microscopy used is microscope or magnifier.The ovum operculum of plant bug ovum part is dyed redness in the plant tissue, and is obvious with the colour contrast of plant tissue, thus the quantity of observed and recorded plant bug ovum.
Said eosin solution is with distilled water or the preparation of 70%-75% alcohol, and the eosin solution for preparing is stored in 4 ℃ of refrigerators.
The present invention also provides a kind of colouring method of plant bug ovum, and it is that vegetable material impregnated in the 0.5-2w/v% eosin solution after 1-5 minute, takes out the water flushing, blots surface moisture then.
The plant bug ovum comes from green plant bug (Apolygus lucorum), black striped plant bug (Adelphocoris suturalis), adelphocoris taeniophorus (Adelphocoris fasiaticollis), alfalfa plant bug (Adelphocoris lineolatus) and/or tarnished plant bug (Lygus pratensis).
The invention provides a kind of dyeing and microscopy method of plant bug ovum, study and observe and predict investigation for quick, the accurately investigation statistics, and related science of plant bug ovum and lay the foundation, be suitable for applying.The dyeing detection method of plant bug ovum of the present invention is applicable to all host plants of plant bug and the histoorgan of each kind of plant.
Description of drawings
Fig. 1 is the Color of coloured differently agent to the green plant bug ovum, a left side: eosin; In: phenol red; Right: safranine.
Fig. 2 is the eosin solution Color of plant bug ovum on the cotton plants different tissues organ, a left side: petiole; In: bract; Right: vein.
Embodiment
Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.
The various coloring agents of embodiment are to the stain test of plant tissue
Supply totally 11 kinds of trial dyeing toners, concrete kind and compound method are followed successively by: it is 0.1%, 0.2%, 1.0%, 1.0%, 9.0% solution that aniline blue, malachite green, safranine T, basic fuchsin, crystal violet directly use distilled water to be mixed with concentration successively; It is 0.3%, 1% solution that phenol red, eosin W or W S use 75% alcohol to be mixed with concentration; It is 0.1% solution that bromcresol purple uses 20% alcohol to be mixed with concentration; It is 0.1% solution that Coomassie brilliant blue R250 uses 3.75% alcohol to be mixed with concentration; It is 0.05% solution that bromophenol blue uses the 0.003mol/L sodium hydroxide solution to be mixed with concentration; The above-mentioned solution in preparation back all can be preserved subsequent use down at 4 ℃.And after the sodium chloride of eriochrome black T and 100 times of weights carries out mixed grinding, directly seal low temperature to preserve, the adding distil water dissolving gets final product during use.Mentioned reagent is all available from the Beijing Chemical Plant.
Get the coloring agent solution 10mL for preparing and add in the test tube, have the beanpod of green plant bug ovum to immerse coloring agent solution product, behind the 3min beanpod is taken out, the flowing water flushing is blotted surperficial water droplet with thieving paper again.The beanpod that is colored is placed under the biological anatomical lens observes, select to the green plant bug ovum painted dark, to the more shallow coloring agent of plant tissue dyeing.Take the common host plants of kind more than 50 such as weeds, herbage, crop, fruit tree from the field, take off cane separately, be placed on respectively in the insect box and supply the green plant bug adult to lay eggs, in 24 hours with its taking-up.Be placed on the cane that has ovum in the coloring agent solution of preliminary screening and dye, do further screening.
Dye one by one to producing Different Organs such as blade that the green plant bug ovum is arranged, flower, flower bud, bell, stem, bract on the cotton plants, relatively the Color of Different Organs ovum.
To the processing of dyeing of green plant bug, black striped plant bug, alfalfa plant bug, adelphocoris taeniophorus and tarnished plant bug ovum on the beanpod, relatively to the Color of five kinds of plant bug ovum.
Be provided with 0.5,1,1.5,2,2.5,3, seven times of 15min handle, coloured differently Color of plant bug ovum on the beanpod under the time relatively, screening optimum dyeing time.
The result shows, the blue and bromcresol purple of aniline does not have coloration basically to the plant bug ovum, and the Color of chrome black and Coomassie brilliant blue is also relatively poor, and safranine, phenol red, eosin, basic fuchsin, crystal violet, malachite green are more satisfactory to the Color of ovum.But dyeing is darker equally with plant tissue for basic fuchsin, crystal violet, malachite green, and colour contrast is not obvious between ovum and the surrounding tissue; And phenol red, eosin, safranine dyeing back plant tissue is painted more shallow, colour contrast apparent in view (Fig. 1) between ovum and the surrounding tissue.
The result shows product is had the branch of 50 various plants of ovum in phenol red, eosin, three kinds of dyeing liquors of safranine, dye one by one, and the result shows: the Color of eosin is best.The Color of eosin is superior to phenol red, and the latter's painted integral body is shallow partially.And safranine is to the painted eosin that is deeper than of ovum surrounding tissue, and the colour contrast of its ovum and plant is not as eosin.
Eosin all has more satisfactory Color (Fig. 2) to plant bug ovum on the Different Organs such as cotton leaf, flower, flower bud, bell, stem, bract.Eosin does not have significant difference to the dyeing efficient basically identical of green plant bug, black striped plant bug, alfalfa plant bug, adelphocoris taeniophorus and tarnished plant bug ovum.The Color of eosin all improves constantly along with the prolongation of dyeing time.The optimum dyeing time of eosin is 2min.
(major diameter is the 0.25-0.40 millimeter to plant bug ovum operculum ovalize; Minor axis is the 0.10-0.16 millimeter), there is slight depression the centre, and general painted darker around the ovum operculum of dyeing back, middle color is shallow partially.Simultaneously, ovum operculum or be higher than plant surface a little, or concordant with plant surface but have obvious gap with plant tissue all around.Through above-mentioned characteristic, can be effectively plant bug ovum operculum and plant tissue or the difference of other insect be come.
The dyeing detection method of plant bug ovum provided by the invention, plant bug ovum are after eosin dyeing, and general naked eyes just can effectively be discerned; Under biological microscope or hand magnifier, the investigation counting is then convenient, accurate.
Though, the present invention has been done detailed description in the preceding text with general explanation and specific embodiments, on basis of the present invention, can to some modifications of do or improvement, this will be apparent to those skilled in the art.Therefore, these modifications or the improvement on the basis of not departing from spirit of the present invention, made all belong to the scope that requirement of the present invention is protected.
Claims (7)
1. the dyeing detection method of a plant bug ovum is characterized in that, vegetable material impregnated in the 0.5-2w/v% eosin solution after 1-5 minute, takes out the water flushing, blots surface moisture then, detects.
2. method according to claim 1 is characterized in that, vegetable material impregnated in the 1w/v% eosin solution 2 minutes.
3. method according to claim 1 and 2 is characterized in that, said eosin solution is with distilled water or the preparation of 70%-75% alcohol.
4. method according to claim 1 and 2 is characterized in that, detects to be visual inspection or microscopy counting.
5. method according to claim 4 is characterized in that, what microscopy used is microscope or magnifier.
6. method according to claim 1 and 2; It is characterized in that the plant bug ovum comes from green plant bug (Apolygus lucorum), black striped plant bug (Adelphocoris suturalis), adelphocoris taeniophorus (Adelphocoris fasiaticollis), alfalfa plant bug (Adelphocoris lineolatus) and/or tarnished plant bug (Lygus pratensis).
7. the colouring method of a plant bug ovum is characterized in that, vegetable material impregnated in the 0.5-2w/v% eosin solution after 1-5 minute, takes out the water flushing, blots surface moisture then.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201110144283A CN102323201A (en) | 2011-05-31 | 2011-05-31 | Staining detection method of eggs of blind stinkbug |
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| CN201110144283A CN102323201A (en) | 2011-05-31 | 2011-05-31 | Staining detection method of eggs of blind stinkbug |
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| CN102323201A true CN102323201A (en) | 2012-01-18 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021036368A1 (en) * | 2019-08-28 | 2021-03-04 | 昆氏(深圳)生物科技有限公司 | Epidermis fungus sampling and staining method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5997847A (en) * | 1998-01-07 | 1999-12-07 | Spiesel; Sydney Z. | Ectoparasite detection method |
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- 2011-05-31 CN CN201110144283A patent/CN102323201A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5997847A (en) * | 1998-01-07 | 1999-12-07 | Spiesel; Sydney Z. | Ectoparasite detection method |
Non-Patent Citations (2)
| Title |
|---|
| 《植物保护科技创新与发展--中国植物保护学会2008年学术年会论文集》 20081231 陆宴辉 等 盲蝽蟓卵的染色技术 第340页 1-7 , * |
| 陆宴辉 等: "盲蝽蟓卵的染色技术", 《植物保护科技创新与发展——中国植物保护学会2008年学术年会论文集》, 31 December 2008 (2008-12-31), pages 340 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021036368A1 (en) * | 2019-08-28 | 2021-03-04 | 昆氏(深圳)生物科技有限公司 | Epidermis fungus sampling and staining method |
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Application publication date: 20120118 |