CN101482520A - Chemical positioning method for tree plant catechin histiocyte - Google Patents
Chemical positioning method for tree plant catechin histiocyte Download PDFInfo
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- CN101482520A CN101482520A CNA2009101161086A CN200910116108A CN101482520A CN 101482520 A CN101482520 A CN 101482520A CN A2009101161086 A CNA2009101161086 A CN A2009101161086A CN 200910116108 A CN200910116108 A CN 200910116108A CN 101482520 A CN101482520 A CN 101482520A
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- acid solution
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- 238000000034 method Methods 0.000 title claims abstract description 17
- 239000000126 substance Substances 0.000 title claims abstract description 11
- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-catechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 title claims description 19
- 235000005487 catechin Nutrition 0.000 title claims description 19
- ADRVNXBAWSRFAJ-UHFFFAOYSA-N catechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3ccc(O)c(O)c3 ADRVNXBAWSRFAJ-UHFFFAOYSA-N 0.000 title claims description 19
- 229950001002 cianidanol Drugs 0.000 title claims description 19
- 210000003701 histiocyte Anatomy 0.000 title claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 74
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 claims abstract description 37
- 239000000463 material Substances 0.000 claims abstract description 23
- 206010020649 Hyperkeratosis Diseases 0.000 claims abstract description 13
- 244000269722 Thea sinensis Species 0.000 claims description 44
- 238000004043 dyeing Methods 0.000 claims description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
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- 241000196324 Embryophyta Species 0.000 claims description 10
- 240000003553 Leptospermum scoparium Species 0.000 claims description 10
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- 235000006468 Thea sinensis Nutrition 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 2
- 239000002253 acid Substances 0.000 abstract description 8
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- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 abstract description 6
- 235000012141 vanillin Nutrition 0.000 abstract description 6
- 230000035945 sensitivity Effects 0.000 abstract description 4
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 abstract description 3
- 210000000056 organ Anatomy 0.000 abstract description 3
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- 150000002989 phenols Chemical class 0.000 abstract 2
- 238000005773 Enders reaction Methods 0.000 abstract 1
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- 239000003086 colorant Substances 0.000 description 11
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- 239000011630 iodine Substances 0.000 description 5
- 229910052740 iodine Inorganic materials 0.000 description 5
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- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 3
- XFZJEEAOWLFHDH-UHFFFAOYSA-N (2R,2'R,3R,3'R,4R)-3,3',4',5,7-Pentahydroxyflavan(48)-3,3',4',5,7-pentahydroxyflavan Natural products C=12OC(C=3C=C(O)C(O)=CC=3)C(O)CC2=C(O)C=C(O)C=1C(C1=C(O)C=C(O)C=C1O1)C(O)C1C1=CC=C(O)C(O)=C1 XFZJEEAOWLFHDH-UHFFFAOYSA-N 0.000 description 2
- KRHYYFGTRYWZRS-UHFFFAOYSA-N Fluorane Chemical compound F KRHYYFGTRYWZRS-UHFFFAOYSA-N 0.000 description 2
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- CWEZAWNPTYBADX-UHFFFAOYSA-N Procyanidin Natural products OC1C(OC2C(O)C(Oc3c2c(O)cc(O)c3C4C(O)C(Oc5cc(O)cc(O)c45)c6ccc(O)c(O)c6)c7ccc(O)c(O)c7)c8c(O)cc(O)cc8OC1c9ccc(O)c(O)c9 CWEZAWNPTYBADX-UHFFFAOYSA-N 0.000 description 2
- MOJZMWJRUKIQGL-FWCKPOPSSA-N Procyanidin C2 Natural products O[C@@H]1[C@@H](c2cc(O)c(O)cc2)Oc2c([C@H]3[C@H](O)[C@@H](c4cc(O)c(O)cc4)Oc4c3c(O)cc(O)c4)c(O)cc(O)c2[C@@H]1c1c(O)cc(O)c2c1O[C@@H]([C@H](O)C2)c1cc(O)c(O)cc1 MOJZMWJRUKIQGL-FWCKPOPSSA-N 0.000 description 2
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- HVQAJTFOCKOKIN-UHFFFAOYSA-N flavonol Natural products O1C2=CC=CC=C2C(=O)C(O)=C1C1=CC=CC=C1 HVQAJTFOCKOKIN-UHFFFAOYSA-N 0.000 description 2
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- 210000000473 mesophyll cell Anatomy 0.000 description 2
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- HGVVOUNEGQIPMS-UHFFFAOYSA-N procyanidin Chemical compound O1C2=CC(O)=CC(O)=C2C(O)C(O)C1(C=1C=C(O)C(O)=CC=1)OC1CC2=C(O)C=C(O)C=C2OC1C1=CC=C(O)C(O)=C1 HGVVOUNEGQIPMS-UHFFFAOYSA-N 0.000 description 2
- 229920002414 procyanidin Polymers 0.000 description 2
- 210000003813 thumb Anatomy 0.000 description 2
- CQNPVMCASGWEHM-VMPITWQZSA-N (e)-3-[4-(dimethylamino)phenyl]prop-2-enoic acid Chemical compound CN(C)C1=CC=C(\C=C\C(O)=O)C=C1 CQNPVMCASGWEHM-VMPITWQZSA-N 0.000 description 1
- YEDFEBOUHSBQBT-UHFFFAOYSA-N 2,3-dihydroflavon-3-ol Chemical compound O1C2=CC=CC=C2C(=O)C(O)C1C1=CC=CC=C1 YEDFEBOUHSBQBT-UHFFFAOYSA-N 0.000 description 1
- KVYRCBOUKXJXDK-UHFFFAOYSA-N 3,4-dimethylphenazine-1,2-diamine hydrochloride Chemical compound Cl.C1=CC=CC2=NC3=C(C)C(C)=C(N)C(N)=C3N=C21 KVYRCBOUKXJXDK-UHFFFAOYSA-N 0.000 description 1
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 description 1
- DQFBYFPFKXHELB-UHFFFAOYSA-N Chalcone Natural products C=1C=CC=CC=1C(=O)C=CC1=CC=CC=C1 DQFBYFPFKXHELB-UHFFFAOYSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical class CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 1
- 108700023158 Phenylalanine ammonia-lyases Proteins 0.000 description 1
- 241000183024 Populus tremula Species 0.000 description 1
- ZONYXWQDUYMKFB-UHFFFAOYSA-N SJ000286395 Natural products O1C2=CC=CC=C2C(=O)CC1C1=CC=CC=C1 ZONYXWQDUYMKFB-UHFFFAOYSA-N 0.000 description 1
- 239000004902 Softening Agent Substances 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 235000005513 chalcones Nutrition 0.000 description 1
- 239000012916 chromogenic reagent Substances 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- QDOXWKRWXJOMAK-UHFFFAOYSA-N dichromium trioxide Chemical compound O=[Cr]O[Cr]=O QDOXWKRWXJOMAK-UHFFFAOYSA-N 0.000 description 1
- PXLWOFBAEVGBOA-UHFFFAOYSA-N dihydrochalcone Natural products OC1C(O)C(O)C(CO)OC1C1=C(O)C=CC(C(=O)CC(O)C=2C=CC(O)=CC=2)=C1O PXLWOFBAEVGBOA-UHFFFAOYSA-N 0.000 description 1
- QGGZBXOADPVUPN-UHFFFAOYSA-N dihydrochalcone Chemical compound C=1C=CC=CC=1C(=O)CCC1=CC=CC=C1 QGGZBXOADPVUPN-UHFFFAOYSA-N 0.000 description 1
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- 150000002213 flavones Chemical class 0.000 description 1
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- 208000014674 injury Diseases 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- MEFBJEMVZONFCJ-UHFFFAOYSA-N molybdate Chemical compound [O-][Mo]([O-])(=O)=O MEFBJEMVZONFCJ-UHFFFAOYSA-N 0.000 description 1
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Abstract
The invention relates to a tea catechin histocyte chemical positioning method, which can solves the problems of the present phenols tissue chemical positioning technique that: bad coloration specificity, slow coloration speed, not high sensitivity, incapability of tea catechin histocyte chemical positioning. The tea catechin histocyte chemical positioning method comprises the following steps: taking the ender tea fresh leaves or tender stalk or root tip or tea callus tissue; producing the slice; dropping 1%-3%(w/v)vanillin concentrated hydrochloric acid solution at one side of the tissue slice and sucking the solution at another side of the tissue slice using filter paper until the material is fully reddened; sucking the redundant vanillin concentrated hydrochloric acid solution using filter paper and drying the slice surface using a blower to obtain a colored slice; taking photomicrograph of the colored slice in 30 minutes after coloration and recording the shape of the colored slice. The coloration process is simple and the operation is convenient. Compared with the typic phenols substance such as ferric chloride, the vanillin acid solution has features of high coloration speed, high sensitivity, good specificity; not only the catechine in the tea organ tissue is positioned and but also the catechine in tea organelle is accurately positioned.
Description
Technical field
The present invention relates to chemical positioning method for tree plant catechin histiocyte.
Background technology
Flavonoids in the plant includes chalcone, flavones, flavonols, flavanols, anthocyanidin, procyanidin etc., and the catechin in the tea tree belongs to the flavanol compound material in the flavonoids.
The coloring agent that is used at present the flavonoid class Histochemical localization both at home and abroad mainly contains Fe
3+Or Fe
2+Molysite, Cr
2O
7 2-, dimethyl diaminophenazine chloride, methylene blue, toluidine, molybdate, vanillin acid solution, butanols sulfuric acid dimethylamino cinnamic acid etc., most of coloring agents all are the broad spectrum activity coloring agents, lack specificity.
Though vanillin acid solution can develop the color with dihydrochalcone, flavanols, flavanonol, procyanidin or condensed tannin reaction, vanillic aldehyde concentrated hydrochloric acid reagent still is widely used in carrying out the detection by quantitative of tea tree flavanols (catechin) content
[1], its reason is that the aldehydes matter content in the tea tree is quite high, and its content can reach bright leaf and the heavy 18-36% of tender stem, and wherein catechin accounts for the 70-80% of Tea Polyphenols.
The person that has the foreign study is used for vanillic aldehyde concentrated hydrochloric acid reagent the report of Histochemical localization of the aldehydes matter of other plant, but because the defective of staining technique as carry out the fixing of material before dyeing, causes aldehydes matter to ooze out, cause Color relatively poor, the phenomenon of color and luster disperse (Fig. 1)
[2]Therefore, can only carry out the Primary Location of organ and the tissue of plant.Do not find that as yet the method is used for the cytology location of aldehydes matter.Do not find the report of the experimental technique of tree plant catechin Histochemical localization as yet.
Summary of the invention
To have in the existing aldehydes matter Histochemical localization technology in order solving that coloring agent lacks that specificity, coloring agent sensitivity are not high, to carry out the defective such as fixing of material before the dyeing, the invention provides a kind of method at tree plant catechin histiocyte chemistry location, this method has colour developing high specificity, easy to operate, characteristics that color speed is fast, highly sensitive.
The technical solution that realizes above-mentioned purpose is as follows:
Chemical positioning method for tree plant catechin histiocyte comprises following operation steps:
A, draw materials
Get bright leaf or tender stem or the tip of a root or the tea callus of the tender tea tree of children, after clear water cleans up, put into clear water, standby;
B, film-making
With fresh leaves of tea plant or tender stem or the tip of a root or the section of cleaning of tea callus;
C, section are taken a photograph
With tableted fresh leaves of tea plant or tender stem or the tip of a root or tea callus photomicrograph picture;
D, dyeing
With side dropping 1%～3% (w/v) vanillic aldehyde concentrated hydrochloric acid solution of dropper, from the opposite side filter paper draw solution of section, until reddening fully to material from section; Draw unnecessary vanillic aldehyde concentrated hydrochloric acid solution with filter paper, and dry up surperficial vanillic aldehyde concentrated hydrochloric acid solution, get stained with hair-dryer;
E, microscopic examination
Dyeed back 30 minutes in, to the stained photomicrograph, record stained dyeing back form.
The present invention compared with prior art has the advantage of following several respects:
1, need not carry out the fixing of material before the present invention dyes, dyeing course is simple, and is easy to operate.
2, at the catechin of tea tree, compare with the developer of the aldehydes matters of using always such as ferric trichloride, vanillin acid solution coloring agent color speed is fast, and is highly sensitive, and specificity is good, sees Table 1.
3, the present invention can not only can also accurately locate tea cell within a cell device catechin tea histoorgan catechin location.
Description of drawings
Fig. 1 be white poplar leaf 20% (w/v) vanillic aldehyde concentrated hydrochloric acid colored graph (select from: Yu-Ying Kao (Gao Yuying) .2002),
Fig. 2 is figure before the bright leaf lower epidermis dyeing of tea,
Fig. 3 is the bright leaf lower epidermis of tea concentrated hydrochloric acid dyeing back figure,
Fig. 4 is the bright leaf lower epidermis of tea 1% vanillic aldehyde concentrated hydrochloric acid dyeing back figure,
Fig. 5 is the bright leaf lower epidermis of tea 3% vanillic aldehyde concentrated hydrochloric acid dyeing back figure,
Fig. 6 is the bright leaf lower epidermis of tea 0.5% vanillic aldehyde concentrated hydrochloric acid dyeing back figure,
Fig. 7 schemes behind the bright leaf lower epidermis of the tea 1% vanillic aldehyde concentrated hydrochloric acid dyeing 10min,
Fig. 8 schemes behind the bright leaf lower epidermis of the tea 1% vanillic aldehyde concentrated hydrochloric acid dyeing 30min,
Fig. 9 schemes behind the bright leaf lower epidermis of the tea 1% vanillic aldehyde concentrated hydrochloric acid dyeing 60min,
Figure 10 is the fixing back of the tender stem FAA of tea 1% a vanillic aldehyde concentrated hydrochloric acid colored graph,
Figure 11 is the tender stem 1% vanillic aldehyde concentrated hydrochloric acid colored graph of tea,
Figure 12 is tea callus cell 1% vanillic aldehyde concentrated hydrochloric acid and iodine liquid dual colored graph.
Embodiment
Below in conjunction with accompanying drawing, the present invention is further described by embodiment.
Embodiment:
Major equipment
1. cutting edge blade (Japan-US board); 2. microslide; 3. cover glass; 4. dropper; 5. diameter 9cm double dish; 6. sharp mouth tweezers; 7. digit microscope.
Material and reagent
1. material:
(1) the bright leaf of tea in the week, tender stem have just been grown in spring or summer; (2) the tea tip of a root that has just grown; (3) tea callus; (4) fresh carrot.
2. main agents:
(1) 1% vanillic aldehyde concentrated hydrochloric acid solution: get the 1g vanillic aldehyde and be dissolved in the 100ml concentrated hydrochloric acid;
(2) clear water;
(3) 10% hydrochloric acid solutions: get 26g concentrated hydrochloric acid concentration (36%) and add 10g water;
(4) softening agent: (50% alcohol: glycerine=1: 1), that is: 50% alcoholic solution and glycerine mixed in by volume 1: 1;
(5) 15% hydrofluorite: get the dense hydrofluoric acid concentration of 32g (47%) and add 15g water;
(6) FAA immobile liquid: formaldehyde by volume: glacial acetic acid: ethanol=1:1:18 mixes;
(7) 0.01mol/L iodine liquid: take by weighing 1.3g iodine, add 2.5g potassium iodide and 25ml distilled water;
(8) 70% sulfuric acid: get 28g concentrated sulphuric acid concentration (98%) and add 70g water.
Method of operating
Staining procedure: draw materials → film-making → dyeing → microscopic examination
1, draws materials
Get the tender tea tree organ of children, promptly fresh bright leaf or tender stem or the tip of a root or the tea callus of tea tree after clear water cleans up, put into clear water, in order to avoid wilt, standby.
2, film-making
Adopt free-hand slicing method.
(1) tealeaves sheet film-making: get fresh carrot, remove xylem, be cut into square section 0.5cm*0.5cm, the preserved radish strip of length 5cm at preserved radish strip one end rip cutting one seam, is cut into suitable size with the bright leaf of tea, is clipped in the slit.
When section, left-handed thumb and forefinger, middle finger are clamped holder, and thumb should be lower than forefinger 2～3mm, in order to avoid cut by blade.Holder will stretch out the outer about 2～3mm of forefinger, and left hand takes holder to want the degree of tightness appropriateness, and the right hand is arrested blade reposefully and become vertical with holder.The edge of a knife is inwardly faced toward holder, and make blade and holder otch keeping parallelism basically, right-handed again arm strength (not using the wrist power of hand) from the left front to right back draw cut equably, gripping object and material are wherein thinly sliced simultaneously, after downcutting several pieces continuously, blade is placed in the double dish that fills clear water in advance rocks gently, section promptly floats in the water.After switching to some, can in double dish, select transparent thin slice low power lens observation and inspection.Good section should be thin and more transparent, institutional framework is complete, otherwise to cut into slices again.Choose cut better sliced materials, be placed on the microslide, drip a last clear water, careful covered is avoided the generation of bubble.Draw unnecessary water with filter paper, photomicrograph, form before the record dyeing.
(2) tealeaves epidermis, mesophyll cell film-making: if observe bright leaf mesophyll cell of tea or leaf epidermal cell Color, bright leaf can be cut into the 0.5cm*0.5cm size, put into 10% (w/w) hydrochloric acid solution 10-20min that dissociates, take off slightly with leaf green and be advisable.The blade that takes out after dissociating on a small quantity with tweezers is placed on the microslide, drips a last clear water, and careful covered is avoided the generation of bubble.Knock cover glass gently with tweezers, allow dispersion of materials open.
(3) the tender stem film-making of tea: get fresh carrot, remove xylem, be cut into square section 0.5cm*0.5cm, the preserved radish strip of length 5cm, at preserved radish strip one end rip cutting one seam, in seam, dig a depression similar with the tender stem shape of tea, that size is identical, the tender stem of tea is clipped in the depression.
(4) tea tip of a root film-making: the same step of method (3).
(5) tea callus film-making: the tea callus need not holder, and directly free-hand section gets final product.
3, section is taken a photograph
With tableted fresh leaves of tea plant or tender stem or the tip of a root or tea callus photomicrograph picture.
4, dyeing
With side dropping 1% (w/v) vanillic aldehyde concentrated hydrochloric acid solution of dropper, from the opposite side filter paper draw solution of section, until reddening fully to section from section.Draw unnecessary vanillic aldehyde concentrated hydrochloric acid solution with filter paper, and dry up the vanillic aldehyde concentrated hydrochloric acid solution of slice surface, avoid concentrated hydrochloric acid health and microscopical injury with hair-dryer, stained.
5, microscopic examination
Dyeed back 30 minutes in, to the stained photomicrograph, record stained dyeing back form.
The technical characterstic explanation
1. dyeing specificity
Can catechin, Tea Saponin, flavanone alcohols and flavonols compound be arranged with the material of vanillic aldehyde concentrated hydrochloric acid solution colour developing in the tea tree, but the content of back three in tea tree is low.And table 1 shows the slope of standard curve of back three and the reaction of vanillic aldehyde concentrated hydrochloric acid than catechin little (table 1), and reaction sensitivity is low, so can not form tangible interference.
Though the redness that anthocyanidin and leucoanthocyanidin present under acid condition can be disturbed the chromogenic reaction of catechin and vanillic aldehyde acid solution, anthocyanidin only accounts for 0.01% of dry weight in the tea, and the middle slightly leucoanthocyanidin of new tea only accounts for the 2%-3% of dry weight
[4], this interference can be ignored, and test also confirms, does not present color reaction (Fig. 2, Fig. 3) through the fresh leaves of tea plant after the acid solution effect that does not contain vanillic aldehyde
2. coloring agent concentration
The concentration of vanillic aldehyde concentrated hydrochloric acid solution coloring agent is at 1%～3% (w/v) comparatively suitable (Fig. 4, Fig. 5), and being lower than 0.5% material can not be by fully dyeing (Fig. 6).Acid solution is concentrated hydrochloric acid or 70% sulfuric acid.
3. dyeing retention time
Material is after the dyeing of 1% (w/v) vanillic aldehyde concentrated hydrochloric acid solution coloring agent, and it is the darkest that color reaches at once, and behind the 30min of dyeing back, color begins to take off gradually (Fig. 7, Fig. 8, Fig. 9).Promptly use the glycerine mounting, can not stop and fade.
4. fixing dyeing
Coloring material is difficult for taking the FAA immobile liquid before dyeing (formaldehyde: glacial acetic acid: ethanol=1:1:18) fixing, the material catechin stripping through fixing causes the dyeing disperse, can not accurately judge the position that catechin accumulates in material.After the FAA immobile liquid was fixing, xylem vessel can not develop the color, its hetero-organization colour developing unintelligible (Figure 10) as the tender stem of tea.Without the fixing tender stem of tea of immobile liquid, after the dyeing of 1% (w/v) vanillic aldehyde concentrated hydrochloric acid solution coloring agent, xylem vessel's dyeing is bright-coloured, and other tissue stainings are clear, do not have the phenomenon (Figure 11) of Color disperse.
5. double staining
This developer can with iodine staining agent double staining, promptly material with iodine staining after, use this chromogenic reagent (Figure 12) again.This is to determining whether nucleus has the catechin accumulation, and is most important.Nucleus dyes without coloring agent, is difficult for observation, is yellow behind iodine staining.
The comparison of table 1 vanillic aldehyde hydrochloric acid colour developing
List of references:
[1] clock trailing plants. tea leaf quality physico-chemical analysis [M]. Shanghai: Shanghai science tech publishing house, 1989,278～279.
[2] Gao Yuying, Scott A Harding, Cai Zhongrui. the difference of two different alanine aminonialyase genes in white poplar accumulation condensed tannin and the lignification cell is expressed [J]. plant physiology, 2002,130:796～807 (Yu-Ying Kao, Scott A.Harding, and Chung-Jui Tsai.Differential expression of two distinct phenylalanineammonia-lyase genes in Condensed tannin-accumulating and lignifying cells ofQuaking Aspen[J] .Plant Physiology, 2002,130:796～807).
[4] as if spring dawn. tea biological chemistry [M]. Beijing: Chinese agriculture publishing house, 2003,18～19.
Claims (1)
1, chemical positioning method for tree plant catechin histiocyte is characterized in that comprising following operation steps:
A, draw materials
Get bright leaf or tender stem or the tip of a root or the tea callus of the tender tea tree of children, after clear water cleans up, put into clear water, standby;
B, film-making
With fresh leaves of tea plant or tender stem or the tip of a root or the section of cleaning of tea callus;
C, section are taken a photograph
With tableted fresh leaves of tea plant or tender stem or the tip of a root or tea callus photomicrograph picture;
D, dyeing
With side dropping 1%～3% (w/v) vanillic aldehyde concentrated hydrochloric acid solution of dropper, from the opposite side filter paper draw solution of section, until reddening fully to material from section; Draw unnecessary vanillic aldehyde concentrated hydrochloric acid solution with filter paper, and dry up surperficial vanillic aldehyde concentrated hydrochloric acid solution, get stained with hair-dryer;
E, microscopic examination
Dyeed back 30 minutes in, to the stained photomicrograph, record stained dyeing back form.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104904846A (en) * | 2015-06-30 | 2015-09-16 | 山东省果树研究所 | Method for fast determining deastringency degree of persimmon |
| CN112525876A (en) * | 2020-12-21 | 2021-03-19 | 河南中医药大学 | Method for observing plant leaf epidermis hair quilt by using fluorescence microscope |
| CN113834790A (en) * | 2021-09-07 | 2021-12-24 | 湘潭大学 | Method for detecting camellia oil doping |
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2009
- 2009-01-21 CN CNA2009101161086A patent/CN101482520A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104904846A (en) * | 2015-06-30 | 2015-09-16 | 山东省果树研究所 | Method for fast determining deastringency degree of persimmon |
| CN104904846B (en) * | 2015-06-30 | 2018-02-13 | 山东省果树研究所 | A kind of quick method for determining persimmon deastringent degree |
| CN112525876A (en) * | 2020-12-21 | 2021-03-19 | 河南中医药大学 | Method for observing plant leaf epidermis hair quilt by using fluorescence microscope |
| CN113834790A (en) * | 2021-09-07 | 2021-12-24 | 湘潭大学 | Method for detecting camellia oil doping |
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