CN106198465A - The method observing the deposition of creeping bentgrass leaf tissue callose based on paraffin section and aniline blue fluorescent staining method - Google Patents
The method observing the deposition of creeping bentgrass leaf tissue callose based on paraffin section and aniline blue fluorescent staining method Download PDFInfo
- Publication number
- CN106198465A CN106198465A CN201610422991.1A CN201610422991A CN106198465A CN 106198465 A CN106198465 A CN 106198465A CN 201610422991 A CN201610422991 A CN 201610422991A CN 106198465 A CN106198465 A CN 106198465A
- Authority
- CN
- China
- Prior art keywords
- wax
- ethanol
- dimethylbenzene
- paraffin
- callose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- 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
- G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
Landscapes
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of method observing creeping bentgrass leaf tissue callose deposition based on paraffin section and aniline blue fluorescent staining method, comprise the steps: S1, compounding medicine；S2, experimental technique successively through drawing materials, fix, being dehydrated, transparent, waxdip, embed, cut into slices, open up sheet, dry sheet, dyeing and mounting flow process and make paraffin section；It is placed on fluorescence microscopy Microscopic observation through fluorescent dye dyeing and takes pictures.The present invention utilizes paraffin section and aniline blue fluorescent staining method to observe the deposition of creeping bentgrass leaf tissue callose first, shorten the experimental period of creeping bentgrass paraffin wax flaking greatly, overcome the film-making difficulty that creeping bentgrass blade is little and thin, breach material itself, restriction that environmental factors is brought, can rapidly and efficiently complete paraffin wax flaking process, and finally obtain the fluorescence microscopy observed result of excellent callose.
Description
Technical field
The present invention relates to microscopic tissue sections and the Fast Detection Technique of histochemical components callose, be specifically related to one
The method observing the deposition of creeping bentgrass leaf tissue callose based on paraffin section and aniline blue fluorescent staining method.
Background technology
Callose is that one has 1, linear β-1 of 6 branches, 3 glucosans, the colloid form structure shortened for overstriking after heating.
In plant development process, callose can be found in a lot of places, including cell plates, pollen, pollen tube, seed, leaf with
And the secondary wall etc. in stem, epidermis, sieve plate, plasmodesmata, xylem forming process.Callose has polymerization rapidly and depolymerization
Physics and physiological property.Additionally, as a part for stress, injured plant, suffer pathogenic bacterial infection or by various
Quickly and in large quantities synthesize when coercing, carry out the extraneous mechanical damage of response, various pathogen invades the biotic caused, and
The biotic etc. that physics, chemistry, envirment factor etc. cause.Under adverse circumstance, callose can be synthesized rapidly and deposit in sieve tube element
Blocking in the surface or sieve aperture of sieve plate, once environmental stimuli releases, and the callose deposited in floor lever of sieve tray or sieve aperture then can
Rapidly disappear, make screen casing recover normal transportation function.Therefore, callose plays important regulation work in the vital movement of plant
With.
Creeping bentgrass is herbaceos perennial, is mainly used in the places such as turfgrass, golf course, football pitch, but
It is in the face of the threat of multiple diseases in growth course, avenges mould for silver dollar pinta, brown spot, compacted spore fungus diseases, cotton maize ear rot, ash
Snow mold sick, pink all infects.When plant suffers pathogen infection, callus mass-energy is quickly accumulated in some intercellular passage
On, pathogen is limited in a certain region, stops it to further expand.And as reply, pathogen also can pass through interactions between protein
Motion-impeding callose is decomposed by the callose digestive enzyme raising host, recovers its motor capacity.Therefore, monitoring crawls to cut
In the case of stock grain husk is caught an illness, the synthesis of callose in vascular bundle and content thereof are to weigh creeping bentgrass to catch an illness situation and screening
Go out the key index with stronger disease resistant plant.It is correlated with currently for callus Quality Research in creeping bentgrass leaf tissue
Be reported in and domestic there is no pertinent literature.
Paraffin section is method classical in histology's conven tional tabletting techniques, that be most widely used.But the paraffin of routine
Dicing method is the longest, is fixed to mounting from drawing materials and makes the time that permanent sheet specimen at least needs continuous one week, and in reality
Application is limited by factors, and during film-making, is related to the use of some harmful medicines, time long
Between experiment can endanger the healthy of experimenter and cause environmental pollution.The most important thing is because its experiment excessive cycle can not
Detect the upgrowth situation of creeping bentgrass timely and disease incidence is adopted remedial measures timely.
Summary of the invention
For solving the problems referred to above, the invention provides a kind of observation based on paraffin section and aniline blue fluorescent staining method and crawl
The method of the deposition of creeping bentgrass leaf tissue callose, utilizes paraffin section and aniline blue fluorescent staining method to observe Jian that crawls first
The deposition of stock grain husk leaf tissue callose, omnidistance time-consuming 2 days of paraffin wax flaking, contracted short creeping bentgrass paraffin wax flaking greatly
Experimental period, breaches material itself, restriction that environmental factors is brought, can rapidly and efficiently complete paraffin wax flaking process.
For achieving the above object, the technical scheme that the present invention takes is:
The side of the deposition of creeping bentgrass leaf tissue callose is observed based on paraffin section and aniline blue fluorescent staining method
Method, comprises the steps:
S1, carry out the preparation of medicine；
S11, by formalin 10ml, acetic acid 3ml, ethanol 87ml, 5ml glycerol mix homogeneously of 50%, obtain FAA and fix
Liquid；
S12, dehydrated alcohol and dimethylbenzene are hybridly prepared into 1/2 dimethylbenzene with the ratio of 1:1；
S13, weigh 0.1g water-soluble aniline blue and be dissolved in the phosphate buffer that 100ml concentration is 0.07mol/L, pH=7
In, obtain the aniline blue fluorescent dye of 0.1%.
S2, making paraffin section；
S21, quickly take off the creeping bentgrass blade of width about 0.8mm with sharp blade, blade cuts is become length
About about 5mm segment, puts in penicillin bottle, adds the FAA fixative of step S11 gained, fixing 12h；
S22, after FAA is fixing, replace successively in 15%, 30%, 50%, 70%, 85% ethanol and carry out serial dehydration, respectively
Dehydration 20min, is dehydrated 30mim in 95% ethanol, is dehydrated twice, each 20min in dehydrated alcohol；Displacement 1/2 2 the most successively
Toluene, dimethylbenzene carry out transparent, transparent 1h in 1/2 dimethylbenzene, in dimethylbenzene transparent twice, and each 30min is complete to material
Till transparent；
S23, in equipped with the penicillin bottle of gained transparent material, add broken wax twice, add for the first time and dimethylbenzene in bottle
The broken wax of equivalent, puts in electric drying oven with forced convection after temperature 36 DEG C keeps 1h to melt completely to added broken wax, adds liquid in bottle
The broken wax of body half, put into electric drying oven with forced convection adjusts the temperature to 36 DEG C overnight after, adjust the temperature to 42 DEG C, replace 50%
After paraffin keeps 1h, temperature is adjusted to 50 DEG C, after the paraffin of displacement 75% keeps 30min, adjusts the temperature to 60 DEG C, replaces pure successively
Paraffin A, paraffin refined wax B, each 30min of paraffin refined wax C, then use TKY-BMB type paraffin wax embedding to embed, in a wax stone one
Individual material, the size of wax stone is: 0.5cm × 0.5cm × 0.5cm, directly repaiies block, cut into slices after embedded wax stone cooling
Or it is standby to be saved in 4 DEG C of refrigerators；
S24, employing wheel type manual microtome carry out serial section, thickness 10 μm；By HH-6 digital display water bath with thermostatic control pot temperature
Regulation, to 45 DEG C, adds distilled water at 500ml large beaker and puts in water-bath, put in beaker by the wax band cut, treat wax band
After fully deployed, the microscope slide that painting wipes Ovum Gallus domesticus album glycerol stretches in beaker, makes wax band adhere on microscope slide, will be stained with wax band
After microscope slide dries naturally, observe exhibition sheet effect under an optical microscope, take the exhibition effective slice, thin piece of sheet and be placed in electric heating air blast and do
Dry case toasts at 38 DEG C, until slice, thin piece is completely dried；
S25, take dry slice, thin piece, be put into successively respectively added with dimethylbenzene, 1/2 dimethylbenzene, the ethanol of 95%, the second of 85%
Alcohol, the ethanol of 70%, the ethanol of 50%, the ethanol of 30%, phosphate buffer, the vertical dye vat of aniline blue stain of 0.1%
In；Each 5min in dimethylbenzene, 1/2 dimethylbenzene, the ethanol of 95%, the ethanol of 85%, the ethanol of 70%, the ethanol of 50%,
30min in the aniline blue stain of each 5S in the ethanol of 30%, 10min in phosphate buffer, 0.1%；Then by after dyeing
Slice, thin piece takes out, and is placed in 30S in distilled water, after cleaning the impurity on slice, thin piece, crosses 1/2 dimethylbenzene, each 10S of dimethylbenzene, treats that slice, thin piece is done
Neutral gum mounting is used after dry；
S26, by after the slice, thin piece natural air drying of gained, be placed in LeiCaDM6000B fluorescence microscopy Microscopic observation and take pictures.
Wherein, described sharp blade is razor blade or single-edge blade.
Wherein, the concentration of the phosphate buffer in described step S4 is 0.07mol/L, pH=7.
The method have the advantages that
Paraffin section and aniline blue fluorescent staining method is utilized to observe the deposition of creeping bentgrass leaf tissue callose first,
Contracted the experimental period of short creeping bentgrass paraffin wax flaking greatly simultaneously, breaches material itself, environmental factors is brought
Limiting, can rapidly and efficiently complete paraffin wax flaking process, and Color is good, coloration result is more stable.
Detailed description of the invention
In order to make objects and advantages of the present invention clearer, below in conjunction with embodiment, the present invention is carried out further
Describe in detail.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not used to limit this
Bright.
Embodiments provide and observe creeping bentgrass vane group based on paraffin section and aniline blue fluorescent staining method
The method knitting the deposition of callose, it is characterised in that comprise the steps:
S1, carry out the preparation of medicine；
S11, by formalin 10ml, acetic acid 3ml, the ethanol 87ml of 50%, glycerol 5ml mix homogeneously, obtain FAA and fix
Liquid；
S12, dehydrated alcohol and dimethylbenzene are hybridly prepared into 1/2 dimethylbenzene with the ratio of 1: 1；
S13, weigh 0.1g water-soluble aniline blue and be dissolved in the phosphate buffer that 100ml concentration is 0.07mol/L, pH=7
In, obtain the aniline blue fluorescent dye of 0.1%.
S2, making paraffin section；
S21, quickly take off the creeping bentgrass blade of diameter about 0.8mm with sharp blade, blade cuts is become length
The segment of about about 5mm, puts in penicillin bottle, adds the FAA fixative of step S11 gained, fixing 12h；
S22, after FAA is fixing, replace successively in the ethanol of 15%, 30%, 50%, 70%, 85%, 95%, 100%
Row serial dehydration, is respectively dehydrated 20min, is dehydrated 30mim, is dehydrated twice, each 30min in dehydrated alcohol in 95% ethanol；Then
Replace 1/2 dimethylbenzene successively, dimethylbenzene carries out transparent, in 1/2 dimethylbenzene transparent twice, each 1h, transparent 1h in dimethylbenzene,
To material fully transparent；
S23, in equipped with the penicillin bottle of gained transparent material, add broken wax twice, add for the first time and dimethylbenzene in bottle
The broken wax of equivalent, puts in electric drying oven with forced convection after temperature 36 DEG C keeps 1h to melt completely to added broken wax, adds liquid in bottle
The broken wax of body half, put into electric drying oven with forced convection adjusts the temperature to 36 DEG C overnight after, adjust the temperature to 42 DEG C, replace 50%
After paraffin keeps 1h, temperature is adjusted to 50 DEG C, after the paraffin of displacement 75% keeps 30min, adjusts the temperature to 60 DEG C, replaces pure successively
Paraffin A, paraffin refined wax B, each 30min of paraffin refined wax C, then use TKY-BMB type paraffin wax embedding to embed, in a wax stone one
Individual material, the size of wax stone is: 0.5cm × 0.5cm × 0.5cm, directly repaiies block, cut into slices after embedded wax stone cooling
Or it is standby to be saved in 4 DEG C of refrigerators；
S24, employing wheel type manual microtome (KD-2508 Kedi Instrument Co., Ltd.) are cut continuously
Sheet, thickness 10 μm；By HH-6 digital display thermostat water bath (Jintan City is along Instrument Ltd. of China) temperature regulation to 45 DEG C (less than stone
Wax fusing point 15 DEG C), add distilled water at 500ml large beaker and put in water-bath, the wax band cut is put in beaker, treats wax band
After fully deployed, the microscope slide that painting wipes Ovum Gallus domesticus album glycerol stretches in beaker, makes wax band adhere on microscope slide, will be stained with wax band
After microscope slide dries naturally, under optical microscope (Motic B3 Professional Series), observe exhibition sheet effect, take
The slice, thin piece opening up sheet effective is placed in electric drying oven with forced convection at 36 DEG C baking, until slice, thin piece is completely dried；
S25, take dry slice, thin piece, be put into successively respectively added with dimethylbenzene, 1/2 dimethylbenzene, the ethanol of 95%, the second of 85%
Alcohol, the ethanol of 70%, the ethanol of 50%, the ethanol of 30%, phosphate buffer, the vertical dye vat of aniline blue stain of 0.1%
In；Each 5min in dimethylbenzene, 1/2 dimethylbenzene, the ethanol of 95%, the ethanol of 85%, the ethanol of 70%, the ethanol of 50%,
30min in the aniline blue stain of each 5S in the ethanol of 30%, 10min in phosphate buffer, 0.1%；Then by after dyeing
Slice, thin piece takes out, and is placed in 30S in distilled water, after cleaning the impurity on slice, thin piece, crosses 1/2 dimethylbenzene, each 10S of dimethylbenzene, treats that slice, thin piece is done
Neutral gum mounting is used after dry；
S26, by after the slice, thin piece natural air drying of gained, be placed in LeiCaDM6000B fluorescence microscopy Microscopic observation and take pictures,
After taking pictures under fluorescence microscope, in photo, pistac speck is callose.
Described sharp blade is razor blade or single-edge blade.The concentration of the phosphate buffer in described step S4 is
0.07mol/L, pH=7.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For Yuan, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (3)
1. the deposition process of creeping bentgrass leaf tissue callose is observed based on paraffin section and aniline blue fluorescent staining method, its
It is characterised by, comprises the steps:
S1, the preparation of medicine；
S11, by formalin 10ml, acetic acid 3ml, ethanol 87ml, 5ml glycerol mix homogeneously of 50%, obtain FAA fixative；
S12, dehydrated alcohol and dimethylbenzene are hybridly prepared into 1/2 dimethylbenzene with the ratio of 1: 1；
S13, weigh 0.1g water-soluble aniline blue and be dissolved in the phosphate buffer that 100ml concentration is 0.07mol/L, pH=7,
The aniline blue fluorescent dye of 0.1%.
S2, making paraffin section；
S21, quickly take off the creeping bentgrass blade of width about 0.8mm with sharp blade, blade cuts is become length about 5mm
The segment of left and right, puts in penicillin bottle, adds the FAA fixative of step S11 gained, fixing 12h；
S22, after FAA is fixing, replace successively and the ethanol of 15%, 30%, 50%, 70%, 85% carry out serial dehydration, each de-
Water 20min, is dehydrated 30mim in 95% ethanol, is dehydrated twice, each 20min in dehydrated alcohol；Replace 1/2 diformazan the most successively
Benzene, dimethylbenzene carry out transparent, transparent 1h in 1/2 dimethylbenzene, in dimethylbenzene transparent twice, and each 30min, until material is complete
Till transparent；
S23, in equipped with the penicillin bottle of gained transparent material, add broken wax twice, add for the first time and dimethylbenzene equivalent in bottle
Broken wax, put in electric drying oven with forced convection after temperature 36 DEG C keeps 1h to melt completely to added broken wax, add liquid one in bottle
The broken wax of half, put into electric drying oven with forced convection adjusts the temperature to 36 DEG C overnight after, adjust the temperature to 42 DEG C, replace 50% paraffin
After keeping 1h, temperature is adjusted to 50 DEG C, after the paraffin of displacement 75% keeps 30min, adjusts the temperature to 60 DEG C, replaces paraffin refined wax successively
A, paraffin refined wax B, each 30min of paraffin refined wax C, TKY-BMB type paraffin wax embedding embeds, a material, wax stone in a wax stone
Size be: 0.5cm × 0.5cm × 0.5cm, after embedded wax stone cooling, directly repair block, carry out cutting into slices or be saved in 4 DEG C
Refrigerator is standby；
S24, employing wheel type manual microtome carry out serial section, thickness 10 μm；HH-6 digital display water bath with thermostatic control pot temperature is regulated
To 45 DEG C, add distilled water at 500ml large beaker and put in water-bath, the wax band light cut is faced down and puts in beaker, treat
After wax band is fully deployed, the microscope slide that painting wipes Ovum Gallus domesticus album glycerol stretches in beaker, makes wax band adhere on microscope slide, will be stained with wax
After the microscope slide of band dries naturally, observe exhibition sheet effect under an optical microscope, take the effective slice, thin piece of exhibition sheet and be placed in electric heating drum
Wind drying baker toasts at 38 DEG C, until slice, thin piece is completely dried；
S25, take dry slice, thin piece, be put into successively respectively added with dimethylbenzene, 1/2 dimethylbenzene, the ethanol of 95%, the ethanol of 85%,
The ethanol of 70%, the ethanol of 50%, the ethanol of 30%, phosphate buffer, 0.1% aniline blue stain vertical dye vat in；
In dimethylbenzene, 1/2 dimethylbenzene each 5min, the ethanol of 95%, the ethanol of 85%, the ethanol of 70%, the ethanol of 50%, 30%
30min in the aniline blue stain of each 5S in ethanol, 10min in phosphate buffer, 0.1%；Then the slice, thin piece after dyeing is taken
Go out, be placed in 30S in distilled water, after cleaning the impurity on slice, thin piece, cross 1/2 dimethylbenzene, each 10S of dimethylbenzene, treat that slice, thin piece is adopted after drying
Use neutral gum mounting；
S26, by after the slice, thin piece natural air drying of gained, be placed in LeiCaDM6000B fluorescence microscopy Microscopic observation and take pictures.
The most according to claim 1 observe creeping bentgrass leaf tissue based on paraffin section and aniline blue fluorescent staining method
The method of the deposition of callose, it is characterised in that described sharp blade is razor blade or single-edge blade.
The most according to claim 1 observe creeping bentgrass leaf tissue based on paraffin section and aniline blue fluorescent staining method
The method of the deposition of callose, it is characterised in that the concentration of the phosphate buffer in described step S4 is 0.07mol/L, pH=
7。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610422991.1A CN106198465A (en) | 2016-06-10 | 2016-06-10 | The method observing the deposition of creeping bentgrass leaf tissue callose based on paraffin section and aniline blue fluorescent staining method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610422991.1A CN106198465A (en) | 2016-06-10 | 2016-06-10 | The method observing the deposition of creeping bentgrass leaf tissue callose based on paraffin section and aniline blue fluorescent staining method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106198465A true CN106198465A (en) | 2016-12-07 |
Family
ID=57453389
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610422991.1A Pending CN106198465A (en) | 2016-06-10 | 2016-06-10 | The method observing the deposition of creeping bentgrass leaf tissue callose based on paraffin section and aniline blue fluorescent staining method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106198465A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106718181A (en) * | 2017-02-10 | 2017-05-31 | 北京农学院 | A kind of method of plant identification to root knot nematode resistance |
| CN109211606A (en) * | 2018-09-06 | 2019-01-15 | 吉林省农业科学院 | A kind of fast method for preparing of pears tissue paraffin section de |
| CN109374373A (en) * | 2018-10-09 | 2019-02-22 | 黑龙江省农业科学院牡丹江分院 | A kind of Apple paraffin section fast method for preparing |
| CN111024472A (en) * | 2020-01-13 | 2020-04-17 | 长沙维世尔生物科技有限公司 | Method for sealing autofluorescence background of paraffin tissue section |
| CN113008648A (en) * | 2021-03-01 | 2021-06-22 | 北京林业大学 | Fluorescence microscopy method for observing behavior of peony pollen tube by combining improved paraffin section with aniline blue dyeing method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102620960A (en) * | 2012-02-29 | 2012-08-01 | 浙江省农业科学院 | Leaf sheath tissue section manufacturing method for enabling users to observe callose of rice leaf sheath tissues |
| CN103149189A (en) * | 2013-03-11 | 2013-06-12 | 山东省农业科学院作物研究所 | Method for rapidly detecting wheat tissue callose |
-
2016
- 2016-06-10 CN CN201610422991.1A patent/CN106198465A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102620960A (en) * | 2012-02-29 | 2012-08-01 | 浙江省农业科学院 | Leaf sheath tissue section manufacturing method for enabling users to observe callose of rice leaf sheath tissues |
| CN103149189A (en) * | 2013-03-11 | 2013-06-12 | 山东省农业科学院作物研究所 | Method for rapidly detecting wheat tissue callose |
Non-Patent Citations (3)
| Title |
|---|
| 张雪英等: "毛竹组织荧光染色显微观察方法的研究", 《竹子研究汇刊》 * |
| 李师翁等: "用苯胺蓝压片法观察小孢子和雄配子体发育过程中胼胝质的动态", 《植物学通报》 * |
| 李文龙: "大豆抗花病毒侵染过程中胼胝质沉积的分子调控机理研究", 《中国博士学位论文全文数据库 农业科技辑》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106718181A (en) * | 2017-02-10 | 2017-05-31 | 北京农学院 | A kind of method of plant identification to root knot nematode resistance |
| CN106718181B (en) * | 2017-02-10 | 2020-05-26 | 北京农学院 | Method for identifying resistance of plants to root-knot nematodes |
| CN109211606A (en) * | 2018-09-06 | 2019-01-15 | 吉林省农业科学院 | A kind of fast method for preparing of pears tissue paraffin section de |
| CN109374373A (en) * | 2018-10-09 | 2019-02-22 | 黑龙江省农业科学院牡丹江分院 | A kind of Apple paraffin section fast method for preparing |
| CN111024472A (en) * | 2020-01-13 | 2020-04-17 | 长沙维世尔生物科技有限公司 | Method for sealing autofluorescence background of paraffin tissue section |
| CN113008648A (en) * | 2021-03-01 | 2021-06-22 | 北京林业大学 | Fluorescence microscopy method for observing behavior of peony pollen tube by combining improved paraffin section with aniline blue dyeing method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106198465A (en) | The method observing the deposition of creeping bentgrass leaf tissue callose based on paraffin section and aniline blue fluorescent staining method | |
| CN102967493A (en) | Rapid paraffin sectioning method for plant tissue | |
| Singer et al. | Pollinarium morphology and floral rewards inBrazilian maxillariinae (Orchidaceae) | |
| Guan et al. | Regeneration of somatic hybrids of ginger via chemical protoplast fusion | |
| Mashimo et al. | Egg structure of Zorotypus caudelli Karny (Insecta, Zoraptera, Zorotypidae) | |
| CN102640742A (en) | Improved manufacturing method for plant wireworm permanent slide | |
| CN107167350A (en) | A kind of preparation method of the paraffin section of eggplant rhizome portion tissue | |
| CN105806682A (en) | Method for preparing leguminous plant leaf paraffin section | |
| Lu et al. | Induction of tetraploidy using high temperature exposure during the first zygote division in Populus adenopoda Maxim | |
| CN109164212A (en) | A kind of measuring method using excised leaf identification Chinese rose germplasm heat resistance | |
| WO2022048544A9 (en) | Method for preparing slices of non-decalcified osteochondral tissue, and cell tracing and rnascope combined test method thereof | |
| CN103609439B (en) | Different strain is on the impact of ginseng Hairy root and application method | |
| CN107576552A (en) | A kind of paraffin section colouring method for observing Chinese Rose infection processs | |
| Zhao et al. | Slow-growing pollen-tube of colchicine-induced 2n pollen responsible for low triploid production rate in Populus | |
| CN105334196A (en) | Method for detection of P-gp mediated Rh123 transport in 3D type organ and application thereof | |
| CN103828580B (en) | The resistant to elevated temperatures screening technique of a kind of field test cotton | |
| Prelezov et al. | Species variety and population structure of Mallophaga (Insecta: Phthiraptera) on chickens in the region of Stara Zagora | |
| Dahmer et al. | Chromosome numbers of Jatropha curcas L.: an important agrofuel plant | |
| CN104297222B (en) | Zebrafish embryo alcoholic liver detecting model and construction method and application of zebrafish embryo alcoholic liver detecting model | |
| CN108007755A (en) | A kind of effectively observation tomato root knot megabacterium paraffin section method | |
| Chen et al. | ADVANCES IN RESEARCH OF LONGAN WITCHES'BROOM DISEASE | |
| Campbell | Anatomy and systematics and of Xyridaceae, with special reference to Aratitiyopea Steyerm. and PE Berry | |
| CN103004726B (en) | A kind of method for enticing collection of simple and effective entomopathogenic nematode | |
| Baksh et al. | An architectural model for Eleocharis: morphology and development of Eleocharis cellulosa (Cyperaceae) | |
| Almatar et al. | Identifying the developmental stages and optimizing the sample preparation for anatomical study of Orthosiphon stamineus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161207 |
|
| RJ01 | Rejection of invention patent application after publication |