CN106399499A - Fluorescence in-situ hybridization method for asparagus fern medium-term chromosomes - Google Patents
Fluorescence in-situ hybridization method for asparagus fern medium-term chromosomes Download PDFInfo
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- CN106399499A CN106399499A CN201610826372.9A CN201610826372A CN106399499A CN 106399499 A CN106399499 A CN 106399499A CN 201610826372 A CN201610826372 A CN 201610826372A CN 106399499 A CN106399499 A CN 106399499A
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- C12Q1/6841—In situ hybridisation
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Abstract
The invention provides a fluorescence in-situ hybridization method for asparagus fern medium-term chromosomes. An asparagus fern medium-term chromosome specimen is prepared by a drop-spreading technique, and by the method, a high-quality asparagus fern chromosome preparation can be obtained. On the basis, fluorescein is used for marking a 45 S rDNA or/and 5D rDNA probe; finally, fluorescence in-situ hybridization of the chromosomes is carried out; and after hybridization of the chromosomes by the probe, fluorescence signals can be directly detected, and therefore, the copy number and the position of a targeted DNA fragment on the chromosomes are determined. The asparagus fern chromosomes which are obtained by the preparation and the fluorescence in-situ hybridization method are complete in morphology, high in hybridization specificity and good in repeatability, operation steps are simple, a large amount of time can be saved, and the fluorescence in-situ hybridization method can be used for identification and structure research of the asparagus fern chromosomes, and provides a new path for evolution research and the like of asparagus linn.
Description
Technical field
The invention belongs to molecular and cytogenetic techniques field is and in particular to a kind of fluorescent in situ of asparagus fern metaphase chromosome
Hybridizing method.
Background technology
Asparagus fern, also known as cloud sheet pine, thorn asparagus fern, Yun Zhu, are Liliaceae Asparagus herbaceous perennial vine plants.Asparagus fern leaf
Piece is soft, and bamboo shape is graceful, throughout the year emerald green, is famous indoor potted flower, is also the excellent lining leaf material of small bonsai and cut-flower, tool
There is high ornamental value.Its root can be used as medicine, and can control acute tracheitis, has the function of moistening the lung and relieve the cough.Research mesh to asparagus fern
Before be concentrated mainly on the aspects such as cultivation technique, its cytological research is relatively lagged behind, this sends out mainly due to the seed of asparagus fern
Bud is relatively difficult, and blade is in needle-like, and shoot tip meristem is also relatively difficult to draw materials, and the material of slide sample is preferably prepared in very difficult acquisition
Material, and its cytoplasm is dense, and cell membrane is harder, processes that cell is relatively difficult, and chromosome is less, be difficult to obtain form clear,
Finely disseminated split coil method.Therefore, it is suitable for the chromosome slide sample preparation of asparagus fern and high-resolution karyotyping technology
Have to be developed.
Fluorescence in situ hybridization technique（fluorescence in situ hybridization, FISH）It is according to nucleic acid
Base pair complementarity principle, by the exogenous nucleic acid through mark（Probe）Intuitively navigate to the one kind on chromosome or chromatin
Molecular cytogenetics method.This technology is beginning of the eighties late 1970s in original radioactive in situ hybridization technology
On the basis of a kind of important nonradioactivein situhybridization technology that grows up.With original radioactive in situ hybridization technology phase
Than, this technology have quick, stability safe, easy and simple to handle and repeatability is strong, same slide can detect simultaneously many
The advantages of individual probe.At present, the method has been widely used for physical map structure, detection GMOs and positioning, heterologous dyeing
Matter identification, chromosome aberration detection, genome structure parsing and the aspect such as species affiliation and evolutionary analysis.But,
In FISH technology operating process, chromosome slide sample quality, the method for probe mark and efficiency, probe and chromosome co-variation
Temperature and time all affect experimental result, the quality of such as chromosome slide sample is that can FISH hybridize successful key factor
One of, enzymolysis is not enough or the excessive failure that all can lead to chromosome sectioning；Slide cytoplasm is too dense, and probe is not easy hybridization and meeting
Cause high background；Whether whether pretreatment properly decides chromosome length moderate, if be conducive to hybridizing.The side of probe mark
Method is different, also can affect results of hybridization accordingly.The temperature and time of probe and chromosome co-variation is also important factor, such as
Fruit is total to that denaturation temperature is low or the time is short, and double-strand does not fully open, and will not hybridize nonspecific signal, and co-variation warm in nature spend height or
Overlong time all can lead to chromosome dilatancy, makes experimental result abnormal.FISH technology is used on a lot of plants
Work(, but, yet there are no the report of fluorescence in situ hybridization technique application in asparagus fern.
Content of the invention
Present invention solves the technical problem that there is provided a kind of easy and simple to handle, time saving and energy saving, stability is high with repeatability
The fluorescence in-situ hybridization method of asparagus fern metaphase chromosome, the form of the asparagus fern metaphase chromosome that the method is observed is intact, signal
Clearly.
The present invention is to solve above-mentioned technical problem to adopt the following technical scheme that, a kind of fluorescent in situ of asparagus fern metaphase chromosome
Hybridizing method it is characterised in that：Using asparagus fern metaphase chromosome as target DNA, with corn 45S rDNA or/and 5S rDNA for visiting
Pin carries out asparagus fern chromosome fluorescence in-situ hybridization.
The fluorescence in-situ hybridization method of asparagus fern metaphase chromosome of the present invention is it is characterised in that concretely comprise the following steps：
（1）The preparation of chromosome slide sample, then potted plant asparagus fern control water three days watered, in the next morning 9 watered:
00-10:00 takes the tip of a root, uses N2After O processes 2h, percentage by volume be 90% Acetic Acid Glacil on fix 10min, fixing after spend from
Wash 10min on sub- water-ice, then cut Root apical meristem area, be placed in quality percent by volume and be respectively 1% pectase and 2%
In the mixed liquor of cellulase in 37 DEG C digest 2h, the ethanol purge tip of a root being 70% with percentage by volume after the completion of enzymolysis twice,
Then the tip of a root, vortex suspension cell are smashed to pieces with dissecting needle in 40 μ L ethanol, centrifugation reservation precipitates and adds 30 μ L anhydrous acetic acids,
Draw 5-8 μ L after mixing and drip piece, room temperature carries out microscopy after placing 5min, the good slide of microscopy is put in UV-crosslinked instrument and passed through
120-125mJ/cm2It is standby that the fixing chromosome of process 2min obtains chromosome slide sample；
（2）Corn 45S rDNA or/and 5S rDNA probe mark, following ingredients are separately added into centrifuge tube, corn on ice
45S rDNA or/and 5S rDNA 2 μ g, 10 × nick translation buffer 2 L, Labeled-dNTP 0.5 L,
Non-labeled-dNTPs 2 L, DNA polymerase I 5 L and DNaseI 0.5 L, then deionized water mends to 20
L, processes 2h in 15 DEG C after mixing in PCR instrument, and addition mass concentration is 140ng/ L salmon sperm DNA 175 L, adds
Percentage by volume be 90% ethanol 450 L and molar concentration be 3mol/L sodium acetate 50 L, after mixing precipitate probe 2h
More than, it is subsequently placed in centrifugation rate centrifugal treating 30min of 13000rpm in centrifuge, then with twice of ethanol purge, will sink
Probe room temperature lucifuge 30min formed sediment is dried；
（3）Elute after hybridization and hybridization, in 0.5 μ L step（2）20 L 2 × SSC, 1 × TE are added to obtain in the label probe obtaining
To probe solution, in step（1）The place having cell on the chromosome slide sample obtaining adds probe solution, covers vinyl cover
Slide, then chromosome slide sample is placed in the canister being covered with moist tissue, boiling water bath processes 5min, then will dye
Body slide sample is put in hybridizing box and is hybridized more than 3h in 55 DEG C, then the taking-up of chromosome slide sample is put in 2 × SSC
5min, removes plastic coverslip；
（4）Signal detection, in step（3）The DAPI fluorescence dye that 20 L contain anti-quencher is added on the chromosome slide sample after process
Liquid dyes 10min, covers long cover plate and is detected.
The invention has the advantages that：
（1）On the basis of existing technology, the method being rewatered with first controlling water processes asparagus fern to the present invention, and the asparagus fern tip of a root can be made mitogenetic
Tissue disintegration activity is vigorous, is conducive to obtaining a large amount of division phases；
（2）The present invention adopts N2O pre-processes the asparagus fern tip of a root, and prepares metaphase chromosome slide sample using drop-spreading technique, can
Assist in removing cytoplasm background, make Chromosome spread good, and method is simply it is not necessary to add a cover piece to go the complicated step such as cover plate
Suddenly, pollution-free；
（3）Probe preparation of the present invention using nick-translation, fluorescein is directly tagged on probe, and labeling effciency is high, and rear
Do not need during continuous in situ hybridization plus antibody can direct detection, hybrid specificities can be improved；
（4）In probe and chromosome co-variation, denaturation is carried out using boiling water bath, establish the suitable time, and in co-variation knot
Directly hybridized after bundle, need not be cooled down, easy and simple to handle pollution-free, and the asparagus fern chromosome morphology obtaining is intact, denaturation is complete,
Signaling point is clear, and number is stable；
（5）The present invention adopts 55 DEG C of high temperature renaturation hybridization, and after hybridization, wash-out, using 2 × SSC room temperature elution in short-term, improves specificity
While, simple to operate and save time；
（6）Whole operation step of the present invention is simple, has saved the plenty of time, has been easy to quickly obtain probe on asparagus fern chromosome
Location information；
（7）The present invention clearly demonstrates the chromosome of asparagus fern, and fluorescence signal is clear, and hybrid specificities are high, establishes and is suitable for literary composition
The metaphase chromosome preparation of bamboo and FISH method；
（8）Corn 45S rDNA and 5S rDNA probe have clearly been positioned on asparagus fern metaphase chromosome the present invention, distinguishable
Homologue and nonhomologous chromosome, contribute to karyotyping, for asparagus fern physical chromosomal map spectrum structure, genetic breeding,
Chromosome Identification and structural research, and the Study on Evolution of asparagus plant etc. has important theory and practice meaning.
Brief description
Fig. 1 is 45S rDNA signal site distributed image on asparagus fern metaphase chromosome（The chromosome that A redyes for DAPI, B
For the signal site of Alexa Fluor-488 fluorescein-labeled 45S rDNA, C is composite diagram）；
Fig. 2 is 5S rDNA signal site distributed image on asparagus fern metaphase chromosome（The chromosome that A redyes for DAPI, B is
The signal site of Texas red fluorescein-labeled 5S rDNA, C is composite diagram）；
Fig. 3 is 45S rDNA and 5S rDNA signal site distributed image on asparagus fern metaphase chromosome simultaneously（A redyes for DAPI
Chromosome, B be Alexa Fluor-488 fluorescein-labeled 45S rDNA signal, C be Texas red fluorescein mark
5S rDNA signal, D be A, B and C composite diagram）.
Specific embodiment
By the following examples the above of the present invention is described in further details, but this should not be interpreted as this
The scope inventing above-mentioned theme is only limitted to below example, all belongs to this based on the technology that the above of the present invention is realized
Bright scope.
Embodiment
（1）The preparation of chromosome slide sample, then potted plant asparagus fern control water three days water, early second day watering
Upper 9:00-10:00 takes the tip of a root, uses N2After O processes 2h, mass percent be 90% Acetic Acid Glacil on fix 10min, fixing after use
Deionized water washes 10min on ice, then cuts Root apical meristem area, is placed in quality percent by volume and is respectively 1% pectase
Digest 2h, the ethanol purge tip of a root being 70% with percentage by volume after the completion of enzymolysis with the mixed liquor of 2% cellulase in 37 DEG C
Twice, then in 40 μ L ethanol, the tip of a root, vortex suspension cell are smashed to pieces with dissecting needle, centrifugation reservation precipitates and adds 30 μ L anhydrous
Acetic acid, draws 5-8 μ L and drips piece, room temperature carries out microscopy after placing 5min, and the good slide of microscopy is put in UV-crosslinked instrument after mixing
Through 120-125mJ/cm2It is standby that the fixing chromosome of process 2min obtains chromosome slide sample；
（2）Corn 45S rDNA or/and 5S rDNA probe mark, following ingredients are separately added into centrifuge tube, corn on ice
45S rDNA or/and 5S rDNA 2 μ g, 10 × nick translation buffer 2 L, Labeled-dNTP 0.5 L,
Non-labeled-dNTPs 2 L, DNA polymerase I 5 L and DNaseI 0.5 L, then deionized water mends to 20
L, processes 2h in 15 DEG C after mixing in PCR instrument, and addition mass concentration is 140ng/ L salmon sperm DNA 175 L, adds
Percentage by volume be 90% ethanol 450 L and molar concentration be 3mol/L sodium acetate 50 L, after mixing precipitate probe 2h
More than, it is subsequently placed in centrifugation rate centrifugal treating 30min of 13000rpm in centrifuge, then with twice of ethanol purge, will sink
Probe room temperature lucifuge 30min formed sediment is dried；
（3）Elute after hybridization and hybridization, in 0.5 μ L step（2）20 L 2 × SSC, 1 × TE are added to obtain in the label probe obtaining
To probe solution, in step（1）The place having cell on the chromosome slide sample obtaining adds probe solution, covers vinyl cover
Slide, then chromosome slide sample is placed in the canister being covered with moist tissue, boiling water bath processes 5min, then will dye
Body slide sample is put in hybridizing box and is hybridized more than 3h in 55 DEG C, then the taking-up of chromosome slide sample is put in 2 × SSC
5min, removes plastic coverslip；
（4）In step（3）The DAPI fluorescence dye liquor dyeing that 20 L contain anti-quencher is added on the chromosome slide sample after process
10min, covers long cover plate（22mm×40mm）Detected.With Olympus BX63 fluorescence microscope, CCD takes pictures,
Metamorph software carries out chromosome analysis, and Photoshop carries out image procossing, and result as shown in Figure 1, Figure 2 and Figure 3, is passed through
Fluorescence microscope is observed that bright hybridization signal.
Embodiment above describes general principle, principal character and the advantage of the present invention, the technical staff of the industry should
Understand, the present invention is not restricted to the described embodiments, the simply explanation present invention's described in above-described embodiment and specification is former
Reason, under the scope without departing from the principle of the invention, the present invention also has various changes and modifications, and these changes and improvements each fall within
In the scope of protection of the invention.
Claims (2)
1. a kind of fluorescence in-situ hybridization method of asparagus fern metaphase chromosome it is characterised in that：Using asparagus fern metaphase chromosome as target
DNA, carries out asparagus fern chromosome fluorescence in-situ hybridization with corn 45S rDNA or/and 5S rDNA for probe.
2. the fluorescence in-situ hybridization method of asparagus fern metaphase chromosome according to claim 1 is it is characterised in that concrete steps
For：
（1）The preparation of chromosome slide sample, then potted plant asparagus fern control water three days watered, in the next morning 9 watered:
00-10:00 takes the tip of a root, uses N2After O processes 2h, percentage by volume be 90% Acetic Acid Glacil on fix 10min, fixing after spend from
Wash 10min on sub- water-ice, then cut Root apical meristem area, be placed in quality percent by volume and be respectively 1% pectase and 2%
In the mixed liquor of cellulase in 37 DEG C digest 2h, the ethanol purge tip of a root being 70% with percentage by volume after the completion of enzymolysis twice,
Then the tip of a root, vortex suspension cell are smashed to pieces with dissecting needle in 40 μ L ethanol, centrifugation reservation precipitates and adds 30 μ L anhydrous acetic acids,
Draw 5-8 μ L after mixing and drip piece, room temperature carries out microscopy after placing 5min, the good slide of microscopy is put in UV-crosslinked instrument and passed through
120-125mJ/cm2It is standby that the fixing chromosome of process 2min obtains chromosome slide sample；
（2）Corn 45S rDNA or/and 5S rDNA probe mark, following ingredients are separately added into centrifuge tube, corn on ice
45S rDNA or/and 5S rDNA 2 μ g, 10 × nick translation buffer 2 L, Labeled-dNTP 0.5 L,
Non-labeled-dNTPs 2 L, DNA polymerase I 5 L and DNaseI 0.5 L, then deionized water mends to 20
L, processes 2h in 15 DEG C after mixing in PCR instrument, and addition mass concentration is 140ng/ L salmon sperm DNA 175 L, adds
Percentage by volume be 90% ethanol 450 L and molar concentration be 3mol/L sodium acetate 50 L, after mixing precipitate probe 2h
More than, it is subsequently placed in centrifugation rate centrifugal treating 30min of 13000rpm in centrifuge, then with twice of ethanol purge, will sink
Probe room temperature lucifuge 30min formed sediment is dried；
（3）Elute after hybridization and hybridization, in 0.5 μ L step（2）20 L 2 × SSC, 1 × TE are added to obtain in the label probe obtaining
To probe solution, in step（1）The place having cell on the chromosome slide sample obtaining adds probe solution, covers vinyl cover
Slide, then chromosome slide sample is placed in the canister being covered with moist tissue, boiling water bath processes 5min, then will dye
Body slide sample is put in hybridizing box and is hybridized more than 3h in 55 DEG C, then the taking-up of chromosome slide sample is put in 2 × SSC
5min, removes plastic coverslip；
（4）Signal detection, in step（3）The DAPI fluorescence dye that 20 L contain anti-quencher is added on the chromosome slide sample after process
Liquid dyes 10min, covers long cover plate and is detected.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106896008A (en) * | 2017-04-21 | 2017-06-27 | 广东省生物工程研究所（广州甘蔗糖业研究所） | A kind of preparation method of spot thatch plant root tip meristematic zone chromosome specimen |
| CN107557491A (en) * | 2017-10-20 | 2018-01-09 | 四川农业大学 | The fluorescence in-situ hybridization method of zanthoxylum armatum metaphase chromosome |
| CN108977565A (en) * | 2018-08-14 | 2018-12-11 | 南京林业大学 | A kind of probe combinations and method suitable for plant chromosome rDNA physical positioning |
| CN109161581A (en) * | 2018-09-06 | 2019-01-08 | 南京晓庄学院 | A kind of Allium mongolicum Regel chromosome fluorescence in-situ hybridization method |
| CN114540534A (en) * | 2021-03-17 | 2022-05-27 | 浙江农林大学 | Mao bamboo oligonucleotide probe |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103305614A (en) * | 2013-06-06 | 2013-09-18 | 北京林业大学 | Lagerstroemia plant chromosomal in-situ hybridization method |
| CN103409524A (en) * | 2013-08-12 | 2013-11-27 | 南开大学 | Fluorescence in situ hybridization method for positioning 45S rDNA on plant chromosome |
| CN103409523A (en) * | 2013-08-12 | 2013-11-27 | 南开大学 | Fluorescence in situ hybridization method of 5S rDNA on plant chromosome |
| CN104073568A (en) * | 2014-07-18 | 2014-10-01 | 西南大学 | Fluorescence in situ hybridization method for metaphase chromosome of mulberry |
-
2016
- 2016-09-18 CN CN201610826372.9A patent/CN106399499A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103305614A (en) * | 2013-06-06 | 2013-09-18 | 北京林业大学 | Lagerstroemia plant chromosomal in-situ hybridization method |
| CN103409524A (en) * | 2013-08-12 | 2013-11-27 | 南开大学 | Fluorescence in situ hybridization method for positioning 45S rDNA on plant chromosome |
| CN103409523A (en) * | 2013-08-12 | 2013-11-27 | 南开大学 | Fluorescence in situ hybridization method of 5S rDNA on plant chromosome |
| CN104073568A (en) * | 2014-07-18 | 2014-10-01 | 西南大学 | Fluorescence in situ hybridization method for metaphase chromosome of mulberry |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106896008A (en) * | 2017-04-21 | 2017-06-27 | 广东省生物工程研究所（广州甘蔗糖业研究所） | A kind of preparation method of spot thatch plant root tip meristematic zone chromosome specimen |
| CN107557491A (en) * | 2017-10-20 | 2018-01-09 | 四川农业大学 | The fluorescence in-situ hybridization method of zanthoxylum armatum metaphase chromosome |
| CN108977565A (en) * | 2018-08-14 | 2018-12-11 | 南京林业大学 | A kind of probe combinations and method suitable for plant chromosome rDNA physical positioning |
| CN109161581A (en) * | 2018-09-06 | 2019-01-08 | 南京晓庄学院 | A kind of Allium mongolicum Regel chromosome fluorescence in-situ hybridization method |
| CN114540534A (en) * | 2021-03-17 | 2022-05-27 | 浙江农林大学 | Mao bamboo oligonucleotide probe |
| CN114540534B (en) * | 2021-03-17 | 2023-08-25 | 浙江农林大学 | Moso bamboo oligonucleotide probe |
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Application publication date: 20170215 |