CN107345873A - A kind of colouring method of improved Tomato Root System root knot and pieces of an egg - Google Patents
A kind of colouring method of improved Tomato Root System root knot and pieces of an egg Download PDFInfo
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- CN107345873A CN107345873A CN201710754053.6A CN201710754053A CN107345873A CN 107345873 A CN107345873 A CN 107345873A CN 201710754053 A CN201710754053 A CN 201710754053A CN 107345873 A CN107345873 A CN 107345873A
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- root system
- sodium hypochlorite
- colouring method
- root
- tomato root
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- 238000000034 method Methods 0.000 title claims abstract description 22
- 235000007688 Lycopersicon esculentum Nutrition 0.000 title claims abstract description 21
- 238000004040 coloring Methods 0.000 title claims abstract description 14
- 240000003768 Solanum lycopersicum Species 0.000 title description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000005708 Sodium hypochlorite Substances 0.000 claims abstract description 19
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims abstract description 19
- 238000005562 fading Methods 0.000 claims abstract description 18
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000000243 solution Substances 0.000 claims abstract description 10
- RZUBARUFLYGOGC-MTHOTQAESA-L acid fuchsin Chemical compound [Na+].[Na+].[O-]S(=O)(=O)C1=C(N)C(C)=CC(C(=C\2C=C(C(=[NH2+])C=C/2)S([O-])(=O)=O)\C=2C=C(C(N)=CC=2)S([O-])(=O)=O)=C1 RZUBARUFLYGOGC-MTHOTQAESA-L 0.000 claims abstract description 8
- 238000007654 immersion Methods 0.000 claims abstract description 8
- 235000011187 glycerol Nutrition 0.000 claims abstract description 7
- 239000007864 aqueous solution Substances 0.000 claims abstract description 5
- 238000005406 washing Methods 0.000 claims abstract description 5
- 241000227653 Lycopersicon Species 0.000 claims abstract 9
- 239000012153 distilled water Substances 0.000 claims description 11
- 239000011159 matrix material Substances 0.000 claims description 4
- 230000000050 nutritive effect Effects 0.000 claims description 4
- 239000008367 deionised water Substances 0.000 claims description 3
- 229910021641 deionized water Inorganic materials 0.000 claims description 3
- 238000011010 flushing procedure Methods 0.000 claims description 2
- 238000007796 conventional method Methods 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 238000011017 operating method Methods 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 13
- 235000013601 eggs Nutrition 0.000 description 13
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 8
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 6
- 238000013461 design Methods 0.000 description 6
- 229910052708 sodium Inorganic materials 0.000 description 6
- 239000011734 sodium Substances 0.000 description 6
- 241000244206 Nematoda Species 0.000 description 5
- 239000000975 dye Substances 0.000 description 5
- AXDJCCTWPBKUKL-UHFFFAOYSA-N 4-[(4-aminophenyl)-(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methyl]aniline;hydron;chloride Chemical compound Cl.C1=CC(=N)C(C)=CC1=C(C=1C=CC(N)=CC=1)C1=CC=C(N)C=C1 AXDJCCTWPBKUKL-UHFFFAOYSA-N 0.000 description 4
- 238000004043 dyeing Methods 0.000 description 4
- 229960000583 acetic acid Drugs 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 229940052223 basic fuchsin Drugs 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 241000243785 Meloidogyne javanica Species 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- WZRZTHMJPHPAMU-UHFFFAOYSA-L disodium;(3e)-3-[(4-amino-3-sulfonatophenyl)-(4-amino-3-sulfophenyl)methylidene]-6-imino-5-methylcyclohexa-1,4-diene-1-sulfonate Chemical compound [Na+].[Na+].C1=C(S([O-])(=O)=O)C(=N)C(C)=CC1=C(C=1C=C(C(N)=CC=1)S([O-])(=O)=O)C1=CC=C(N)C(S(O)(=O)=O)=C1 WZRZTHMJPHPAMU-UHFFFAOYSA-L 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 230000002792 vascular Effects 0.000 description 2
- 241000380130 Ehrharta erecta Species 0.000 description 1
- 241001677344 Meloidogyne ethiopica Species 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000000442 meristematic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 229960005076 sodium hypochlorite Drugs 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Preparation Of Fruits And Vegetables (AREA)
Abstract
The present invention provides the colouring method of a kind of improved Tomato Root System root knot and pieces of an egg, including：1) Tomato Root System is first used into aqueous sodium hypochlorite solution immersion treatment, washing removes the sodium hypochlorite of residual；2) Tomato Root System after being handled in step 1) is immersed in the acid fuchsin aqueous solution, sealed, low temperature is placed；3) Tomato Root System after being handled in step 2) is taken out, is rinsed and faded with water, the sample after colour fading is put into glycerine and preserved, you can.For conventional method, improved method of the invention have the advantages that operating procedure it is easy, it is time-consuming it is short, fading effect is good.
Description
Technical field
The present invention relates to the improvement of a kind of Tomato Root System root knot and pieces of an egg colouring method.
Background technology
Tomato originates in the ground such as Ecuador, Peru, Victoria earliest, is Solanaceae tomato genus, annual or perennial grass
This plant.Until 19th century people just have found that tomato is edible and nutritious.It is then just universal all over the world
Plantation, one of important fruit type vegetables in China are turned into till now.
Root knot nematode disease endangers tomato serious.Root-knot nematode second instar larvae (J2) enters from the position close to elongation zone plants
Thing root, moved to vascular bundle, after finding suitable position, 5-7 cell near head is pierced through and by esophageal gland with lancet
Secretion injects, and induces these cells to form giant cell.With the formation of giant cell, the root tissue hyperplasia near nematode increases
Thickness, so that the root outwardly convex of plant, can thus form root knot.Nematode mainly infects plant lateral roots and fibrous root, forms string
The small root knot of shape, initial stage is yellow-white, sub-translucent, and smooth matter is soft, and the later stage is brown and coarse perishable, influences tomato plant
Normal growth.
J2 enters plant tissue from close to meristematic zone position, and the center pillar towards the tip of a root moves.Along with the hair of root-tip cells
Educate and extend, it can move to the maturation zone of root, after finding suitable feeding site, settle down and bud into third-instar larvae
(J3) giant cell that 4-5 parenchyma cell forms large-scale multinuclear, that metabolism is active of head, and gradual shape, are stimulated
Into the vascular tissue for being adapted to oneself living needs, in order to which it absorbs plant nutrient from giant cell, until development is adult.
Male worm can leave feeding site, and female adult can lay eggs.Pieces of an egg may proceed to hatch the root tissue for into J2, infecting plant again.In host
During plant and the long-term interaction of nematode, host plant has some defensive measures to resist nematode invasion or destroy nematode
Development.
In experiment, usually count plant root knot number and pieces of an egg number come illustrate the resistance of plant pair root-knot nematode power.For
Root knot number and pieces of an egg number are more conveniently counted, it is necessary to dyed to root knot and pieces of an egg, conventional method is sodium hypochlorite-acid
Property moral training method, but the acid fuchsine solution of conventional method is formulated and dyed by basic fuchsin, acetic acid and distilled water
It is required to boil with when fading, fading time is longer.
The content of the invention
It is an object of the invention to provide a kind of improved Tomato Root System root knot and the colouring method of pieces of an egg.
The colouring method of Tomato Root System root knot provided by the present invention and pieces of an egg, comprises the steps：
1) Tomato Root System is first used into aqueous sodium hypochlorite solution immersion treatment, washing removes the sodium hypochlorite of residual；
2) Tomato Root System after being handled in step 1) is immersed in the acid fuchsin aqueous solution, sealed, low temperature is placed；
3) Tomato Root System after being handled in step 2) is taken out, is rinsed and faded with water, the sample after colour fading is put into glycerine
Middle preservation, you can.
The above method takes out before the operation in carrying out step 1), in addition to by Tomato Root System from nutritive cube, is used in combination
Running water cleans up root system to operation of the surface without matrix.
In above method step 1), in the aqueous sodium hypochlorite solution, the mass concentration of sodium hypochlorite can be 1.0%-
1.5%.
The time of the immersion treatment can be 3-5min.
The concrete operations of sodium hypochlorite that the washing removes residual are：First by after aqueous sodium hypochlorite solution immersion treatment
Root system rinses 30s with running water, is then transferred in distilled water and soaks 15min.
In above method step 2), the concentration of the acid fuchsin aqueous solution is to contain acid product per in 1000mL distilled water
Red 3-4g, concretely 3.5g.
The temperature that the low temperature is placed can be 0-4 DEG C, and concretely 4 DEG C, the time can be 8-12 hours, and concretely 8 is small
When.
In above method step 3), the rinse water concretely deionized water.
The time of the flushing can be 3-5min, concretely 5min.
Compared with traditional experiment, the method after improvement need not boil during dyeing, colour fading, take short；Examination used
Agent sodium hypochlorite and acid fuchsin are fewer and stingless than agents useful for same in conventional method (sodium hypochlorite, basic fuchsin, glacial acetic acid)
Swash property smell；Fade obvious compared with conventional method, picture shooting effect is good.
Brief description of the drawings
Improved sodium hypochlorite-acid fuchsin dyeing that Fig. 1 is the present invention is dyed with traditional sodium hypochlorite-acid fuchsin
Effect contrast figure.Wherein A is that traditional sodium hypochlorite-acid fuchsin dyes, colour fading 5min design sketch；B is traditional hypochlorous acid
Sodium-acid fuchsin dyeing, colour fading 7d design sketch；C dyes for the improved sodium hypochlorite of the present invention-acid fuchsin, colour fading 5min
Design sketch.
Embodiment
Below by specific embodiment, the present invention will be described, but the invention is not limited in this.
Experimental method used in following embodiments is conventional method unless otherwise specified；Institute in following embodiments
Reagent, material etc., unless otherwise specified, commercially obtain.
Embodiment
Root system is dyed using the improved colouring method of the present invention, concrete operations are：
1. carefully taking out plant root from nutritive cube, root system is cleaned up to surface without matrix with running water；
2. clean root system is placed on after soaking 4min in the liquor natrii hypochloritis of mass concentration 1.5%, rushed with running water
Wash 30s and be transferred in distilled water and soak 15min to remove the sodium hypochlorite of residual；
3. during root system to be transferred to the beaker for filling 100mL distilled water, add 3.5g acid fuchsin；
4. beaker is sealed with sealed membrane, be then placed in 4 DEG C of refrigerators 8 hours；
5. taking sample to go out, faded within 5 minutes with deionized water rinsing；
6. after fading, sample is put into preserve in glycerine and is easy to take pictures and counts.
Fig. 1 C are colour fading 5min design sketch.
Comparative example
Root system is dyed using traditional colouring method, concrete operations are：
Plant root is carefully taken out from nutritive cube, is cleaned up root system to surface without matrix with running water.It will clean
Root system be placed on and soak 4min in the liquor natrii hypochloritis of mass concentration 1.5% after, rinse 30s with running water and be transferred to distilled water
Middle immersion 15min is to remove the sodium hypochlorite of residual.Root system is transferred in the beaker for filling 100ml distilled water, adds 3ml acid
Property fuchsin solution (3.5g basic fuchsin solids are first dissolved in 250ml acetic acid, add 750ml distilled water mix), boil 1-
5min (depending on the situation that the time paints according to root knot, is dyed to red by most root knot and is defined, if being not easy for a long time
Color, a small amount of the acid fuchsine solution can be added into beaker again), the magenta of the residual on root system surface is rinsed after cooling with running water,
It is then transferred into the beaker for filling 100ml acidity glycerine (in glycerine plus a few drop 5mol/L HCl) and heats colour fading 0.5-1h, most
Root is transferred to glycerine afterwards:Lactic acid:Distilled water (1:1:1) in, red is deeper i.e. at visible root knot and pieces of an egg after root fades, now just
Root knot number on root and pieces of an egg number can be counted.
Figure 1A is that traditional sodium hypochlorite-acid fuchsin dyes, colour fading 5min design sketch；
Figure 1B is that traditional sodium hypochlorite-acid fuchsin dyes, colour fading 7d design sketch.
As can be seen from the figure traditional dyeing, after colour fading 5min, DeGrain, root knot and root tissue are all red, are taken off
After color 7 days, root tissue is essentially colourless, and root knot and pieces of an egg are still red, and fading effect is preferable, convenient observation and counting.And use
Colouring method after the improvement of the present invention dyes, and after colour fading 5min, positive effect, root knot and pieces of an egg are dyed to red, root tissue
Substantially colorless, contrast is obvious.
Claims (5)
1. the colouring method of a kind of Tomato Root System root knot and pieces of an egg, comprises the steps：
1) Tomato Root System is first used into aqueous sodium hypochlorite solution immersion treatment, washing removes the sodium hypochlorite of residual；
2) Tomato Root System after being handled in step 1) is immersed in the acid fuchsin aqueous solution, sealed, low temperature is placed；
3) Tomato Root System after being handled in step 2) is taken out, is rinsed and faded with water, the sample after colour fading is put into glycerine and protected
Deposit, you can.
2. colouring method according to claim 1, it is characterised in that：Operation of the methods described in step 1) is carried out it
Before, in addition to Tomato Root System taken out from nutritive cube, and cleaned up root system to operation of the surface without matrix with running water.
3. colouring method according to claim 1 or 2, it is characterised in that：In step 1), the aqueous sodium hypochlorite solution
In, the mass concentration of sodium hypochlorite is 1.0%-1.5%；
The time of the immersion treatment is 3-5min；
The operation of sodium hypochlorite that the washing removes residual is：First the root system after aqueous sodium hypochlorite solution immersion treatment is used certainly
Water rinses 30s, is then transferred in distilled water and soaks 15min.
4. according to the colouring method any one of claim 1-3, it is characterised in that：In step 2), the acid fuchsin
The concentration of the aqueous solution is to contain acid fuchsin 3-4g per in 1000mL distilled water；
The temperature that the low temperature is placed is 0-4 DEG C, and the time is 8-12 hours.
5. according to the colouring method any one of claim 1-4, it is characterised in that：In step 3), the rinse water
For deionized water；
The time of the flushing is 3-5min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710754053.6A CN107345873B (en) | 2017-08-29 | 2017-08-29 | A kind of colouring method of improved Tomato Root System root knot and pieces of an egg |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710754053.6A CN107345873B (en) | 2017-08-29 | 2017-08-29 | A kind of colouring method of improved Tomato Root System root knot and pieces of an egg |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107345873A true CN107345873A (en) | 2017-11-14 |
| CN107345873B CN107345873B (en) | 2019-11-22 |
Family
ID=60257235
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710754053.6A Active CN107345873B (en) | 2017-08-29 | 2017-08-29 | A kind of colouring method of improved Tomato Root System root knot and pieces of an egg |
Country Status (1)
| Country | Link |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102183394A (en) * | 2011-03-08 | 2011-09-14 | 沈阳农业大学 | Method for making hyacinth root tip chromosome slice |
| CN104969782A (en) * | 2015-06-22 | 2015-10-14 | 中国农业科学院郑州果树研究所 | Peach rootstock south root-knot nematode resisting in-vitro rapid screening method |
-
2017
- 2017-08-29 CN CN201710754053.6A patent/CN107345873B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102183394A (en) * | 2011-03-08 | 2011-09-14 | 沈阳农业大学 | Method for making hyacinth root tip chromosome slice |
| CN104969782A (en) * | 2015-06-22 | 2015-10-14 | 中国农业科学院郑州果树研究所 | Peach rootstock south root-knot nematode resisting in-vitro rapid screening method |
Non-Patent Citations (3)
| Title |
|---|
| 张春龙等: "淡紫拟青霉防治小麦禾谷孢囊线虫病研究", 《植物保护》 * |
| 李子龙: "miR319a对番茄生长发育的影响及在番茄抗根结线虫中作用的初探", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 海飞等: "利用植物性添加物防治番茄根结线虫病的研究", 《长江蔬菜》 * |
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| Publication number | Publication date |
|---|---|
| CN107345873B (en) | 2019-11-22 |
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