CN104472331A - Fast transparentizing method for observing fungi of plant roots - Google Patents

Fast transparentizing method for observing fungi of plant roots Download PDF

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Publication number
CN104472331A
CN104472331A CN201410719663.9A CN201410719663A CN104472331A CN 104472331 A CN104472331 A CN 104472331A CN 201410719663 A CN201410719663 A CN 201410719663A CN 104472331 A CN104472331 A CN 104472331A
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plant
root
solution
transparent
plant roots
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CN201410719663.9A
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CN104472331B (en
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唐明
李朕
吴娜
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Northwest A&F University
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Northwest A&F University
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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 fast transparentizing method for observing fungi of plant roots. The method includes preparing mixed solution from components by mass, then adding water to determine volume, and finally mixing the mixed solution with NaClO solution with a concentration from 4%to 8% according to a volume ratio 19 to 1 so as to form transparent solution; collecting fresh plant root samples, cleaning the root samples with flushing water, cutting the root samples into root sections and immersing the root sections into the transparent solution 5 to 10 minutes at a temperature between 10 DEG C to 25 DEG C; flushing the root samples until no transparent solution residue is left and finishing transparentizing of plant roots; performing fungi colonization microscopic observation to the plant roots after transparentized. Plant root cells are wonderfully protected by the transparent solution adopted, cell damage to plant roots is avoided, observation of plant roots and endophytic fungi is benefited, and complex operating process and plenty of waiting time in prior art are decreased. The fast transparentizing method for observing fungi of plant roots has the advantages of being quick, easy and protective to root cells and the like, the transparentizing efficiency and observation effect of plant roots are highly improved.

Description

For the fast transparent method of observation of plant root system fungi
Technical field
The present invention relates to botany, plant symbiosis and parasitic microbe to learn, be specifically related to a kind of fast transparent method for observation of plant root system fungi, belong to biology techniques field.
Background technology
Root system is the organ that plant obtains mineral nutrition and moisture from the media such as soil, learns field, the transparent and dyeing of root system of plant botany, plant symbiosis and parasitic microbe, most important for the growth of root system of plant, microorganism position observation.But simultaneously again can not be long to the clearing time of root system of plant, because the long meeting of clearing time causes damage to root system of plant cell, instead be unfavorable for the truth of better observation of plant root system.
In order to ensure when not damaging root system of plant, again can the real conditions of clearly observation of plant root system inside, researcher expands intensive research, propose the multiple method transparent to root system of plant, mainly containing following two kinds: one for the common method that root system of plant is transparent is at present that root system of plant sample is cut into root segment, then use 5 ~ 10%KOH solution as transparent liquid water-bath in 90 DEG C of water-baths, the transparent liquid renewed when solution colour is darker is until root system is transparent.This kind of clearing method is better for the root system sample effect that the colors such as young root are more shallow, but transparent for the root system that the colors such as the old root of plant are darker, needs the troublesome operation of long period, so larger to the damage of root system of plant; Two is utilize NaClO and HNO 3fast transparent is carried out to root system of plant, although this kind of clearing method carries out transparent can saving time for the old root of the plant that color is darker, but easily damage is caused to root system of plant owing to utilizing the strong oxidizing property of reagent and Bleachability quick decolorization of carrying out, cause the disconnected joint of root system of plant and cell structure imperfect, and the desirable observing effect of root system of plant can not be obtained.
Summary of the invention
The object of the invention is to overcome at present for the drawback existing for root system of plant clearing method, and disclose a kind of fast transparent method for observation of plant root system fungi.
For achieving the above object, the technical solution adopted in the present invention is as follows:
A. by following content of component preparation mixed solution: NaCl 8, KCl 0.2, Na 2hPO 412H 2o1.6 ~ 3.6, KH 2pO 40.2 ~ 0.24, re-use NaOH and HCl and regulate pH in 7.0 ~ 8.0 scopes; Then adding water and be settled to 1000 parts, is finally that the NaClO solution of 4 ~ 8% is mixed into clear solution, temperature 4 DEG C of preservations according to volume/volume than for 19:1 by above-mentioned mixed liquor and concentration;
B. gather fresh plant root sample, running water is clean and be cut into the long root segment of 1 ~ 5cm;
C. plant root segment is immersed in clear solution completely, soak 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. Fungal colonization microexamination is carried out to transparent rear root system of plant.
The present invention adopt clear solution fabulous protect root system of plant cell; avoid the cellular damage that root system of plant is caused; more be conducive to observing root system of plant and endogenetic fungus; decrease the loaded down with trivial details operating process of conventional transparent method and a large amount of stand-by period; gather fast, the advantage such as convenient, protection root cell, 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 contrast figure.
In accompanying drawing, A starts state when adding 0.4%NaClO solution, and B is the state started when adding clear solution, and C adds the state of 0.4%NaClO solution after 10 minutes, and D adds the state of clear solution after 10 minutes.
Accompanying drawing 2 be the present invention be applied to fresh Poplar Roots transparent, dyeing and compressing tablet after microexamination figure.
In accompanying drawing, A is the state using clear solution to observe Poplar Roots fungi, and B is the state using 0.4%NaClO solution to observe root system fungi, and arrow is designated as root system and breaks joint place.
Embodiment
Below in conjunction with embodiment and accompanying drawing, technical scheme of the present invention is described in further detail:
A. by following mass component preparation mixed solution: NaCl 8g, KCl 0.2g, Na 2hPO 412H 2o3.6g, KH 2pO 40.24g, re-uses NaOH and HCl and regulates pH 7.4, add water and be settled to 1L, get 95ml mixed solution and 5ml concentration be 8% NaClO solution be mixed into clear solution, temperature 4 DEG C of preservations;
B. gather fresh poplar root sample, running water is clean, is cut into the long root segment of 5cm;
C. willow root segment is immersed in clear solution completely, soak 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. Fungal colonization microexamination is carried out to transparent rear Poplar Roots: transparent rear Poplar Roots is used 10%H 2o 2softening 2 minutes, 2%HCl acidifying was after 3 minutes, then with the dyeing of trypan blue dye liquor, and with lactic acid and glycerine by volume/volume ratio is that 1:1 decolours and makes compressing tablet afterwards and observe and take pictures.
From accompanying drawing, although transparent effect of the present invention and control group difference are not quite, but clear solution clarification of the present invention, and control group solution is muddy, this root system of plant is caused to the result damaged and cause tissue juice to leak outside in a large number because control group clearing time is long, the root system of control group clearing method presents a large amount of disconnected joint simultaneously, and in the present invention, root system of plant observing effect is better, does not produce damage.

Claims (2)

1., for the fast transparent method of observation of plant root system fungi, it is characterized in that adopted technical scheme is as follows:
A. by following content of component preparation mixed solution: NaCl 8, KCl 0.2, Na 2hPO 412H 2o1.6 ~ 3.6, KH 2pO 40.2 ~ 0.24, re-use NaOH and HCl and regulate pH in 7.0 ~ 8.0 scopes; Then adding water and be settled to 1000 parts, is finally that the NaClO solution of 4 ~ 8% is mixed into clear solution, temperature 4 DEG C of preservations according to volume/volume than for 19:1 by above-mentioned mixed liquor and concentration;
B. gather fresh plant root sample, running water is clean and be cut into the long root segment of 1 ~ 5cm;
C. plant root segment is immersed in clear solution completely, soak 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. Fungal colonization microexamination is carried out to transparent rear root system of plant.
2. the fast transparent method for observation of plant root system fungi according to claim 1, is characterized in that:
A. by following mass component preparation mixed solution: NaCl 8g, KCl 0.2g, Na 2hPO 412H 2o3.6g, KH 2pO 40.24g, re-uses NaOH and HCl and regulates pH 7.4, add water and be settled to 1L, get 95ml mixed solution and 5ml concentration be 8% NaClO solution be mixed into clear solution, temperature 4 DEG C of preservations;
B. gather fresh poplar root sample, running water is clean, is cut into the long root segment of 5cm;
C. willow root segment is immersed in clear solution completely, soak 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. Fungal colonization microexamination is carried out to transparent rear Poplar Roots: transparent rear Poplar Roots is used 10%H 2o 2softening 2 minutes, 2%HCl acidifying was after 3 minutes, then with the dyeing of trypan blue dye liquor, and with lactic acid and glycerine by volume/volume ratio is that 1:1 decolours and makes compressing tablet afterwards and observe and take pictures.
CN201410719663.9A 2014-12-01 2014-12-01 Fast transparent method for observation of plant root system fungus Active CN104472331B (en)

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CN104472331B CN104472331B (en) 2016-11-23

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104996183A (en) * 2015-07-28 2015-10-28 中国农业大学 Method for detecting influence of microbes to plant roots and special device for implementing same
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)

* Cited by examiner, † Cited by third party
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

Patent Citations (6)

* Cited by examiner, † Cited by third party
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

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104996183A (en) * 2015-07-28 2015-10-28 中国农业大学 Method for detecting influence of microbes to plant roots and special device for implementing same
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

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