CN104472331B - Fast transparent method for observation of plant root system fungus - Google Patents
Fast transparent method for observation of plant root system fungus Download PDFInfo
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- CN104472331B CN104472331B CN201410719663.9A CN201410719663A CN104472331B CN 104472331 B CN104472331 B CN 104472331B CN 201410719663 A CN201410719663 A CN 201410719663A CN 104472331 B CN104472331 B CN 104472331B
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- plant
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- root
- clear solution
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N3/00—Preservation of plants or parts thereof, e.g. inhibiting evaporation, improvement of the appearance of leaves or protection against physical influences such as UV radiation using chemical compositions; Grafting wax
Abstract
The invention discloses a kind of fast transparent method for observation of plant root system fungus, be to be configured to mixed solution by content of component, then add water constant volume, with concentration be finally 4～8% NaClO solution be mixed into clear solution according to volume/volume than for 19:1；Gathering fresh plant root sample, flowing water is rinsed well and is cut into root segment and immerses in clear solution, soaks 5～10 minutes temperature 10～25 DEG C；Rinsing and remain to without clear solution, root system of plant is transparent to be completed；Transparent rear root system of plant is carried out Fungal colonization microexamination.What the used clear solution of the present invention was fabulous protects root system of plant cell; avoid the cell injury that root system of plant is caused; it is more beneficial for root system of plant and endogenetic fungus are observed; decrease the loaded down with trivial details operating process of conventional transparent method and substantial amounts of waiting time; gather the advantages such as quick, convenient, protection root cell, greatly improve the transparent efficiency of root system of plant and the observing effect to root system of plant.
Description
Technical field
The present invention relates to botany, plant symbiosis and parasitic microbe learn, be specifically related to a kind of for observing
The fast transparent method of root system of plant fungus, belongs to biology techniques field.
Background technology
Root system is that plant obtains mineral nutrition and the organ of moisture from the media such as soil, at botany, plants
Thing symbiosis and parasitic microbe learn field, the transparent and dyeing of root system of plant, for root system of plant growth,
Microorganism position observation is most important.But simultaneously again can not be long to the clearing time of root system of plant, because
Root system of plant cell is caused damage by the long meeting of clearing time, instead be unfavorable for more preferable observation of plant root system
Truth.
In order to ensure in the case of not damaging root system of plant, again can be clearly within observation of plant root system
Real conditions, research worker expands intensive research, it is proposed that multiple transparent to root system of plant
Method, being currently used for the transparent common method of root system of plant mainly has following two: one is by root system of plant
Sample is cut into root segment, and then use 5～10%KOH solution are as transparent liquid water-bath in 90 DEG C of water-baths,
The transparent liquid renewed when solution colour is deeper is until root system is transparent.This kind of clearing method is for face such as young roots
The shallower root system sample effect of color is preferable, but transparent for root system that the colors such as the old root of plant are deeper, needs relatively
Troublesome operation for a long time, so the damage to root system of plant is bigger；Two is to utilize NaClO and HNO3To planting
Thing root system carries out fast transparent, although this kind of clearing method carries out transparent for the old root of plant that color is deeper
Can save the time, but the strong oxidizing property and the Bleachability quick decolorization carried out owing to utilizing reagent is the most right
Root system of plant causes damage, causes the disconnected joint of root system of plant and cellularity imperfect, and can not obtain plant
The preferable observing effect of root system.
Summary of the invention
It is an object of the invention to overcome the drawback being currently used for existing for root system of plant clearing method, and public
Open a kind of fast transparent method for observation of plant root system fungus.
For achieving the above object, the technical solution adopted in the present invention is as follows:
A. mixed solution: NaCl 8, KCl 0.2, Na are prepared by following content of component2HPO4·12H2O
1.6～3.6, KH2PO40.2～0.24, re-use NaOH Yu HCl regulation pH in 7.0～8.0 scopes；So
After add water and be settled to 1000 parts, finally by above-mentioned mixed liquor and concentration be 4～8% NaClO solution according to
Volume/volume is mixed into clear solution, 4 DEG C of preservations of temperature than for 19:1；
B. gathering fresh plant root sample, 1～5cm long root segment is rinsed and be cut into flowing water well；
C. plant root segment is completely immersed in clear solution, soaks 5～10 minutes temperature 10～25 DEG C；
D. clear water is rinsed well and is remained to without clear solution, and root system of plant is transparent to be completed；
E. transparent rear root system of plant is carried out Fungal colonization microexamination.
What the used clear solution of the present invention was fabulous protect root system of plant cell, it is to avoid to root system of plant
The cell injury caused, is more beneficial for observing root system of plant and endogenetic fungus, decreases tradition thoroughly
Operating process that bright method is loaded down with trivial details and substantial amounts of waiting time, gather quickly, convenient, protection root system is thin
The advantages such as born of the same parents, greatly improve the transparent efficiency of root system of plant and the observing effect to root system of plant.
Accompanying drawing explanation
Accompanying drawing 1 is that the present invention is applied to fresh Poplar Roots transparent effect comparison figure.
State when A is to be initially added into 0.4%NaClO solution in accompanying drawing, when B is for being initially added into clear solution
State, C is to add 0.4%NaClO solution state after 10 minutes, and D is for adding clear solution after 10 minutes
State.
Accompanying drawing 2 be the present invention be applied to fresh Poplar Roots transparent, dyeing and tabletting after microexamination figure.
In accompanying drawing, A is the state using clear solution to observe Poplar Roots fungus, and B is for using 0.4%NaClO
Solution observes the state of root system fungus, and arrow is designated as root system and breaks at joint.
Detailed description of the invention
Below in conjunction with embodiment and accompanying drawing, technical scheme is described in further detail:
A. by following mass component preparation mixed solution: NaCl 8g, KCl 0.2g, Na2HPO4·12H2O
3.6g, KH2PO40.24g, re-use NaOH Yu HCl regulation pH 7.4, add water and be settled to 1L, take 95ml
Mixed solution and 5ml concentration be 8% NaClO solution be mixed into clear solution, 4 DEG C of preservations of temperature；
B. gathering fresh poplar root sample, flowing water is rinsed well, is cut into the long root segment of 5cm；
C. willow root segment is completely immersed in clear solution, soaks 10 minutes temperature 20 DEG C；
D. clear water is rinsed well and is remained to without clear solution, and Poplar Roots is transparent to be completed；
E. transparent rear Poplar Roots is carried out Fungal colonization microexamination: used by transparent rear Poplar Roots
10%H2O2Softening 2 minutes, 2%HCl was acidified after 3 minutes, then dyeed with trypan blue dye liquor, and with lactic acid and
Glycerol by volume/volume ratio be 1:1 decolouring after make tabletting observe and take pictures.
From accompanying drawing, although transparent effect of the present invention is little with matched group difference, but the present invention is transparent molten
Liquid is clarified, and matched group solution is muddy, and this is to cause root system of plant owing to matched group clearing time is long
Damage causes the result that tissue fluid leaks outside in a large number, and the root system of matched group clearing method presents a large amount of disconnected simultaneously
Joint, and in the present invention, root system of plant observing effect is preferable, does not produce damage.
Claims (2)
1. for the fast transparent method of observation of plant root system fungus, it is characterised in that the technical side used
Case is as follows:
A. mixed solution: NaCl 8, KCl 0.2, Na are prepared by following content of component2HPO4·12H2O
1.6～3.6, KH2PO40.2～0.24, re-use NaOH Yu HCl regulation pH in 7.0～8.0 scopes；
Then add water and be settled to 1000 parts, finally by above-mentioned mixed liquor and concentration be 4～8% NaClO solution by
It is mixed into clear solution, 4 DEG C of preservations of temperature than for 19:1 according to volume/volume；
B. gathering fresh plant root sample, 1～5cm long root segment is rinsed and be cut into flowing water well；
C. plant root segment is completely immersed in clear solution, soaks 5～10 minutes temperature 10～25 DEG C；
D. clear water is rinsed well and is remained to without clear solution, and root system of plant is transparent to be completed；
E. transparent rear root system of plant is carried out Fungal colonization microexamination.
Fast transparent method for observation of plant root system fungus the most according to claim 1, it is special
Levy and be:
A. by following mass component preparation mixed solution: NaCl 8g, KCl 0.2g, Na2HPO4·12H2O
3.6g, KH2PO40.24g, re-use NaOH Yu HCl regulation pH 7.4, add water and be settled to 1L, take 95ml
Mixed solution and 5ml concentration be 8% NaClO solution be mixed into clear solution, 4 DEG C of preservations of temperature；
B. gathering fresh poplar root sample, flowing water is rinsed well, is cut into the long root segment of 5cm；
C. willow root segment is completely immersed in clear solution, soaks 10 minutes temperature 20 DEG C；
D. clear water is rinsed well and is remained to without clear solution, and Poplar Roots is transparent to be completed；
E. transparent rear Poplar Roots is carried out Fungal colonization microexamination: used by transparent rear Poplar Roots
10%H2O2Softening 2 minutes, 2%HCl was acidified after 3 minutes, then dyeed with trypan blue dye liquor, and with lactic acid and
Glycerol by volume/volume ratio be 1:1 decolouring after make tabletting observe and take pictures.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410719663.9A CN104472331B (en) | 2014-12-01 | 2014-12-01 | Fast transparent method for observation of plant root system fungus |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410719663.9A CN104472331B (en) | 2014-12-01 | 2014-12-01 | Fast transparent method for observation of plant root system fungus |
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| Publication Number | Publication Date |
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| CN104472331A CN104472331A (en) | 2015-04-01 |
| CN104472331B true CN104472331B (en) | 2016-11-23 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201410719663.9A Active CN104472331B (en) | 2014-12-01 | 2014-12-01 | Fast transparent method for observation of plant root system fungus |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104996183B (en) * | 2015-07-28 | 2018-08-17 | 中国农业大学 | A kind of method and its dedicated unit of influence of the detection microorganism to root system of plant |
| CN109443883A (en) * | 2018-12-28 | 2019-03-08 | 齐鲁师范学院 | It is a kind of handle plant root tip clarifier and its application |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3928136A (en) * | 1972-10-26 | 1975-12-23 | Pierre A Launey | Container for collection of cultures of parasitic fungi and a method of using the same |
| CN1434127A (en) * | 2002-01-22 | 2003-08-06 | 裴芳君 | Staining kit and preparation method, staining method, and use thereof |
| CN102116711A (en) * | 2011-01-31 | 2011-07-06 | 山东东方海洋科技股份有限公司 | Manufacturing method of paraffin sections of zostera marina embryo |
| CN102607907A (en) * | 2012-02-24 | 2012-07-25 | 东北农业大学 | Paraffin section method for fern gametophytes |
| 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 |
-
2014
- 2014-12-01 CN CN201410719663.9A patent/CN104472331B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3928136A (en) * | 1972-10-26 | 1975-12-23 | Pierre A Launey | Container for collection of cultures of parasitic fungi and a method of using the same |
| CN1434127A (en) * | 2002-01-22 | 2003-08-06 | 裴芳君 | Staining kit and preparation method, staining method, and use thereof |
| CN102116711A (en) * | 2011-01-31 | 2011-07-06 | 山东东方海洋科技股份有限公司 | Manufacturing method of paraffin sections of zostera marina embryo |
| CN102607907A (en) * | 2012-02-24 | 2012-07-25 | 东北农业大学 | Paraffin section method for fern gametophytes |
| 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 |
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| CN104472331A (en) | 2015-04-01 |
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