CN102181511A - Method for simultaneously dyeing mycelial masses and starch in mycorhiza of arethusa - Google Patents
Method for simultaneously dyeing mycelial masses and starch in mycorhiza of arethusa Download PDFInfo
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- CN102181511A CN102181511A CN2011100420540A CN201110042054A CN102181511A CN 102181511 A CN102181511 A CN 102181511A CN 2011100420540 A CN2011100420540 A CN 2011100420540A CN 201110042054 A CN201110042054 A CN 201110042054A CN 102181511 A CN102181511 A CN 102181511A
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- mycorhiza
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- dyestuff
- chloral hydrate
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Abstract
The invention discloses a method for simultaneously dyeing mycelial masses and starch in mycorhiza of arethusa, which comprises the following steps: taking a mycorhiza slice, placing on a glass slide, dripping chloral hydrate iodine solution dye, dyeing, then sucking away the redundant dye, further dripping aniline blue lactoglycerin solution dye, dyeing, then sucking away the redundant dye, and placing under an optical microscope for micro-observation, so that the starch in the mycorhiza is in dark brown and the fungal mycelial masses are in blue. By adopting the method, the mycelial masses and the starch in the mycorhiza can be simultaneously dyed, and the colonization characteristics of fungi of the mycorhiza in a root of the arethusa and the starch distribution characteristics can be simultaneously observed, thereby having important significance for revealing the relationship between digestion and formation of the fungi of the mycorhiza and accumulation and the digestion of a host carbohydrate.
Description
Technical field
The present invention relates to mycorhiza staining technique field, particularly relate to a kind of orchid mycorhiza hypha body and starch painted method simultaneously.
Background technology
The long-term symbiosis of orchid and fungi forms the mycorhiza structure, and mycorrhizal fungi is present in the root tegumental cell with the form of hypha body.Mycorrhizal fungi and host's mutual reciprocity and mutual benefit exchange aspect carbohydrate mutually.Therefore from the relation of cytology angle research mycorrhizal fungi and host's carbohydrate, to plant symbiotic relationship extremely important to understanding bacterium.The general iodine liquid that adopts dyes to starch, is coloured to blueness or red-purple, with toluidine blue, C.I. 42685 etc. tela contexta is dyeed.Aspect the plant mycorhiza, C.I. 42685 lactophenol oil, aniline blue etc. are generally used in the dyeing of mycorrhizal fungi.
In certain stage of orchid growth of mycorrhiza, the starch and the hypha body of roots of plants storage are present in the same cell simultaneously, are not easily distinguishable.At present, all be to adopt single stain to the observation of starch and hypha body in this stage, can only dye to mycorrhizal fungi or starch, and simultaneously clear view to hypha body and starch.
Summary of the invention
Technical problem to be solved by this invention is to overcome prior art can not observe the hypha body in the mycorhiza and the defective of starch simultaneously, and a kind of method of can dye simultaneously hypha body and starch is provided.
In order to solve the problems of the technologies described above, the present invention adopts following technical scheme:
Orchid mycorhiza hypha body of the present invention and starch is painted method simultaneously: get the mycorhiza section, place on the slide glass, drip Chloral Hydrate iodine liquid dyestuff, siphon away unnecessary dyestuff after the dyeing, drip aniline blue glycerine lactate buffer solution dyestuff again, siphon away unnecessary dyestuff after the dyeing, place microscopic examination under the opticmicroscope, the starch in the mycorhiza presents dark-brown, hypha,hyphae group presents blueness; Wherein:
Chloral Hydrate iodine liquid dyestuff is formulated by Chloral Hydrate 80～120g, potassiumiodide 4～6g, iodine 1～2g and deionized water 80～120ml;
Aniline blue glycerine lactate buffer solution dyestuff is formulated by lactic acid 8～12ml, glycerine 18～22ml, water-soluble aniline blue 0.03～0.05g and deionized water 8～12ml.
Further, aforementioned dyeing process is: get healthy mycorhiza, clean, section, the mycorhiza sheet is placed on the slide glass, drip 2～3 of Chloral Hydrate iodine liquid dyestuffs, dyeed 8～10 seconds, siphon away unnecessary dyestuff with filter paper, drip 1～2 of aniline blue glycerine lactate buffer solution dyestuff again, dyeed 5～6 seconds, siphon away unnecessary dyestuff with filter paper, place microscopic examination under the opticmicroscope, the starch in the mycorhiza presents dark-brown, hypha,hyphae group presents blueness; Wherein:
Chloral Hydrate iodine liquid dyestuff is formulated by Chloral Hydrate 100g, potassiumiodide 5g, iodine 1.5g and deionized water 100ml;
Aniline blue glycerine lactate buffer solution dyestuff is formulated by put on record 0.04g and deionized water 10ml of lactic acid 10ml, glycerine 20ml, water-soluble this case.
The compound method of used Chloral Hydrate iodine liquid dyestuff is in the aforementioned dyeing process: get potassiumiodide and iodine, put into clean beaker, add deionized water dissolving, add Chloral Hydrate then, dissolving is kept in the brown bottle standby.
The compound method of used aniline blue glycerine lactate buffer solution dyestuff is in the aforementioned dyeing process: water intaking soluble aniline indigo plant, put into clean beaker, and add deionized water dissolving, add lactic acid, glycerine then, it is standby to put into drop bottle after the mixing.
Adopt technique scheme to dye, starch in the orchid mycorhiza can be dyed dark-brown, hypha,hyphae group dyes blueness (as depicted in figs. 1 and 2), if and the dyeing reversed in order or will two kinds staining fluids dye after mixing, hypha body all can not be dyed blueness.
Compared with prior art, technical scheme of the present invention successfully dyes simultaneously to hypha body in the mycorhiza and starch, can observe the distribution characteristics of deciding grow characteristics and starch of mycorrhizal fungi in the orchid root simultaneously, significant to the accumulation that discloses the clearing up of mycorrhizal fungi, formation and host's carbohydrate, the relation cleared up.
Description of drawings
Fig. 1 is amplified to 100 times mycorhiza dyeing photo;
Fig. 2 is amplified to 400 times mycorhiza dyeing photo;
Among the figure, dark-brown is the starch that is colored, and blueness is the hypha body that is colored.
Embodiment
Describe technical scheme of the present invention in detail below in conjunction with embodiment.
At first prepare dyestuff, concrete compound method is as follows:
Chloral Hydrate iodine liquid dyestuff: accurately weighing 1.5g iodine, 5g potassiumiodide, put into that clean beaker grinds and, add 100ml deionized water water dissolution, add the 100g Chloral Hydrate then and mix, dissolving promptly, is kept in the brown bottle standby.
Aniline blue glycerine lactate buffer solution dyestuff: accurate weighing 0.04g water-soluble aniline blue, put into clean beaker, add the 10ml deionized water, stirring and dissolving adds 10ml lactic acid, 20ml glycerine again, mixes, that is and, it is standby to put into drop bottle.
Dyeing process:
1, gets the oyster white root of orchid health, under tap water, clean dust outside the root;
2, with the free-hand slicing method section, choose thin and complete section and be placed on the slide glass standby;
3, dropping 2～3 is dripped and is closed chloral-iodine liquid dyestuff in the section of slide glass, dyes 8～10 seconds, siphons away unnecessary dyestuff with filter paper;
4, in the slide glass section, drip 1～2 of aniline blue glycerine lactate buffer solution dyestuff then, dyeed 5～6 seconds, siphon away unnecessary dyestuff with filter paper;
5, the slide glass that will have a dyeing back section places microscopic examination under the opticmicroscope, can observe clearly that starch in the same cell presents dark-brown and the mycorrhizal fungi hypha body presents blueness, as depicted in figs. 1 and 2.
Claims (4)
1. orchid mycorhiza hypha body and starch painted method simultaneously, it is characterized in that: get the mycorhiza section, place on the slide glass, drip Chloral Hydrate iodine liquid dyestuff, siphon away unnecessary dyestuff after the dyeing, drip aniline blue glycerine lactate buffer solution dyestuff again, siphon away unnecessary dyestuff after the dyeing, place microscopic examination under the opticmicroscope, the starch in the mycorhiza presents dark-brown, hypha,hyphae group presents blueness; Wherein:
Chloral Hydrate iodine liquid dyestuff is formulated by Chloral Hydrate 80～120g, potassiumiodide 4～6g, iodine 1～2g and deionized water 80～120ml;
Aniline blue glycerine lactate buffer solution dyestuff is formulated by lactic acid 8～12ml, glycerine 18～22ml, water-soluble aniline blue 0.03～0.05g and deionized water 8～12ml.
2. according to claim 1 described orchid mycorhiza hypha body and painted method of starch while, it is characterized in that: get healthy mycorhiza, clean, section, the mycorhiza sheet is placed on the slide glass, drip 2～3 of Chloral Hydrate iodine liquid dyestuffs, dyeed 8～10 seconds, siphon away unnecessary dyestuff with filter paper, drip 1～2 of aniline blue glycerine lactate buffer solution dyestuff again, dyeed 5～6 seconds, siphon away unnecessary dyestuff with filter paper, place microscopic examination under the opticmicroscope, the starch in the mycorhiza presents dark-brown, hypha,hyphae group presents blueness; Wherein:
Chloral Hydrate iodine liquid dyestuff is formulated by Chloral Hydrate 100g, potassiumiodide 5g, iodine 1.5g and deionized water 100ml;
Aniline blue glycerine lactate buffer solution dyestuff is formulated by put on record 0.04g and deionized water 10ml of lactic acid 10ml, glycerine 20ml, water-soluble this case.
3. according to claim 1 or 2 described orchid mycorhiza hypha bodies and painted method of starch while, it is characterized in that: the compound method of described Chloral Hydrate iodine liquid dyestuff is: get potassiumiodide and iodine, put into clean beaker, add deionized water dissolving, add Chloral Hydrate then, dissolving is kept in the brown bottle standby.
4. according to claim 1 or 2 described orchid mycorhiza hypha bodies and painted method of starch while, it is characterized in that: the compound method of described aniline blue glycerine lactate buffer solution dyestuff is: water intaking soluble aniline indigo plant, put into clean beaker, add deionized water dissolving, add lactic acid, glycerine then, it is standby to put into drop bottle after the mixing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110042054 CN102181511B (en) | 2011-02-22 | 2011-02-22 | Method for simultaneously dyeing mycelial masses and starch in mycorhiza of arethusa |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201110042054 CN102181511B (en) | 2011-02-22 | 2011-02-22 | Method for simultaneously dyeing mycelial masses and starch in mycorhiza of arethusa |
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| CN102181511A true CN102181511A (en) | 2011-09-14 |
| CN102181511B CN102181511B (en) | 2013-04-10 |
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| CN 201110042054 Expired - Fee Related CN102181511B (en) | 2011-02-22 | 2011-02-22 | Method for simultaneously dyeing mycelial masses and starch in mycorhiza of arethusa |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103411813A (en) * | 2013-05-28 | 2013-11-27 | 江苏大学 | Method for rapidly and efficiently dyeing arbuscular mycorrhizal fungi |
| CN104132940A (en) * | 2014-06-30 | 2014-11-05 | 上海市园林科学研究所 | Convenient observation method of orchid mycorrhiza microstructure |
| CN106323723A (en) * | 2016-11-30 | 2017-01-11 | 宁夏大学 | Double-blue staining method |
| CN115824757A (en) * | 2022-12-28 | 2023-03-21 | 九江学院 | Dyeing method of endophytic fungi in oil tea root system |
-
2011
- 2011-02-22 CN CN 201110042054 patent/CN102181511B/en not_active Expired - Fee Related
Non-Patent Citations (4)
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| E. PIANO ET AL.: "Specificity of Host-Endophyte Association in Tall Fescue Populations from Sardinia, Italy", 《CROP SCIENCE》 * |
| 张卫民, 王振中: "稻瘟病菌与水稻CO39 近等基因系互作的寄生适合度属性研究", 《植物保护》 * |
| 方中达: "《植病研究方法（第三版）》", 31 December 1998, 中国农业出版社 * |
| 陈瑞蕊等: "兰科菌根的研究进展", 《应用与环境生物学报》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103411813A (en) * | 2013-05-28 | 2013-11-27 | 江苏大学 | Method for rapidly and efficiently dyeing arbuscular mycorrhizal fungi |
| CN103411813B (en) * | 2013-05-28 | 2017-02-08 | 江苏大学 | Method for rapidly and efficiently dyeing arbuscular mycorrhizal fungi |
| CN104132940A (en) * | 2014-06-30 | 2014-11-05 | 上海市园林科学研究所 | Convenient observation method of orchid mycorrhiza microstructure |
| CN106323723A (en) * | 2016-11-30 | 2017-01-11 | 宁夏大学 | Double-blue staining method |
| CN106323723B (en) * | 2016-11-30 | 2019-01-22 | 宁夏大学 | Double indigo plant decoration methods |
| CN115824757A (en) * | 2022-12-28 | 2023-03-21 | 九江学院 | Dyeing method of endophytic fungi in oil tea root system |
| CN115824757B (en) * | 2022-12-28 | 2024-03-26 | 九江学院 | Dyeing method for endophytic fungi of camellia oleifera root system |
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| CN102181511B (en) | 2013-04-10 |
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