CN108841989A - A kind of the SSR molecular marker system and its application of muskmelon - Google Patents
A kind of the SSR molecular marker system and its application of muskmelon Download PDFInfo
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- 235000009847 Cucumis melo var cantalupensis Nutrition 0.000 title claims abstract description 25
- 239000003147 molecular marker Substances 0.000 title claims abstract description 16
- 244000064895 Cucumis melo subsp melo Species 0.000 title description 27
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 claims abstract description 11
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 claims abstract description 11
- 244000241257 Cucumis melo Species 0.000 claims abstract 9
- 239000002773 nucleotide Substances 0.000 claims description 12
- 125000003729 nucleotide group Chemical group 0.000 claims description 12
- 238000002372 labelling Methods 0.000 claims description 3
- 238000000034 method Methods 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 2
- 108020004414 DNA Proteins 0.000 description 31
- 108091092878 Microsatellite Proteins 0.000 description 30
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 16
- 229960000935 dehydrated alcohol Drugs 0.000 description 6
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 6
- 229960004756 ethanol Drugs 0.000 description 5
- 235000019441 ethanol Nutrition 0.000 description 5
- 238000001502 gel electrophoresis Methods 0.000 description 5
- 238000001228 spectrum Methods 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 238000012408 PCR amplification Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 3
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- 238000000137 annealing Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
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- 238000006062 fragmentation reaction Methods 0.000 description 2
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- 239000000243 solution Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 208000002109 Argyria Diseases 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
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- 239000003550 marker Substances 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical class OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
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Abstract
The invention discloses a kind of SSR molecular marker system of muskmelon and its applications, are related to molecular marking technique field.The present invention is for identifying that the primer pair title of melon variety is respectively C30, MU5499, MU5554-1, SSR12083 and SSR04219.The above-mentioned SSR molecular marker primer pair polymorphism that the present invention studies is high, band is clear, repeatability is stablized, and can be used for constructing muskmelon DNA fingerprinting and identification melon variety.Especially identify that western state is No. 1 close, western state is No. 3 close, western state is No. 25 close, western state is No. 17 close, western state close No. 21 or western state close 29, effect becomes apparent from.
Description
Technical field
The present invention relates to molecular marking technique field more particularly to the SSR molecular marker systems and its application of a kind of muskmelon.
Background technique
SSR (Simple Sequence Repeats) label is that one kind developed in recent years is with specific primer PCR
The molecular marking technique on basis, also referred to as microsatellite DNA (MicrosatelliteDNA) are a kind of (general by several nucleotide
Be 1~6) be repeat unit group at the tandem repetitive sequence up to tens nucleotide.The sequence of each two sides SSR is general
It is relatively conservative single-copy sequence.
In the genome of biology, a large amount of repetitive sequence is contained especially in the genome of higher organism, the study found that micro-
In satellite there is height and make a variation in the number of recurring unit, these variations show as the ortholoidy variation of microsatellite number or repeat single
Sequence in bit sequence is possible to polymorphism that is not exactly the same, thus causing multiple sites.If these variations can be taken off
It shows and, different SSR can be found in different kinds even interindividual polymorphism.Due to a certain specific in genome
The flanking sequence of microsatellite be usually all the stronger unique sequence of conservative, thus can be by the DNA fragmentation of microsatellite flank
Clone, sequencing, then according to the flanking sequence of microsatellite can artificial synthesized primer carry out PCR amplification, thus will be single micro-
Satellite site, which amplifies, to be come.Due to the variation of single microsatellite locus repetitive unit quantitatively, individual amplified production is in length
Variation on degree just generates the polymorphism of length, this polymorphism is known as simple sequence repeats length polymorphism (SSLP), each
Amplification site just represents a pair of alleles in this site.Compared with other molecular labelings, SSR marker has following excellent
Point：(1) quantity is abundant, covers whole gene group, and the polymorphism of announcement is high；(2) with the characteristic of multiple alleles, the letter provided
Breath amount is high；It (3) is in codominance with Mendelian fashion heredity；(4) each site is determined by the primer sequence designed, convenient for difference
The mutual exchange and cooperation in laboratory develop primer.Thus the technology is widely used in the building of genetic map, target gene at present
Calibration, fingerprint image the research such as drafting in.
Using the technology existing research of SSR molecular marker identification muskmelon, but with greater need for developing, polymorphism is high, band is clear
Clear, repeated stable SSR molecular marker is for constructing muskmelon DNA fingerprinting.
Summary of the invention
In view of this, the embodiment of the invention provides a kind of SSR molecular marker system of muskmelon and its application, main purpose
It is that the SSR molecular marker that solves for identifying melon variety is many kinds of so that melon variety can not be identified fast and accurately
Problem.
In order to achieve the above objectives, invention broadly provides following technical solutions：
On the one hand, the embodiment of the invention provides a kind of SSR molecular marker systems of muskmelon；The molecular labeling system packet
Include 5 primer pairs；
The entitled C30 of primer pair 1, nucleotides sequence are classified as SEQ ID NO.1 and SEQ ID NO.2；
The entitled MU5499 of primer pair 2, nucleotides sequence are classified as SEQ ID NO.3 and SEQ ID NO.4；
The entitled MU5554-1 of primer pair 3, nucleotides sequence are classified as SEQ ID NO.5 and SEQ ID NO.6；
The entitled SSR12083 of primer pair 4, nucleotides sequence are classified as SEQ ID NO.7 and SEQ ID NO.8；
The entitled SSR04219 of primer pair 5, nucleotides sequence are classified as SEQ ID NO.9 and SEQ ID NO.10.
Preferably, the new varieties of the muskmelon be western state is No. 1 close, western state is No. 3 close, western state is No. 25 close, western state is No. 17 close,
Western state close No. 21 or western state close 29.
On the other hand, the embodiment of the invention provides above-mentioned muskmelon SSR molecular marker systems in identification melon variety
Using.
Compared with prior art, the beneficial effects of the invention are as follows：
The present invention is recorded its base sequence information and annealing temperature, is filtered out more by screening muskmelon genome SSR primer
The SSR molecular marker system that state property is high, band is clear, repeatability is stable can be quickly quasi- for constructing muskmelon DNA fingerprinting
Really identify melon variety.
Detailed description of the invention
Fig. 1 is the gel electrophoresis spectrum after the primer 73 that the embodiment of the present invention 2 provides expands；
Fig. 2 is the gel electrophoresis spectrum after the primer 47 that the embodiment of the present invention 2 provides expands；
Fig. 3 is the gel electrophoresis spectrum after the primer 14 that the embodiment of the present invention 2 provides expands；
Fig. 4 is the gel electrophoresis spectrum after the primer 53 that the embodiment of the present invention 2 provides expands；
Fig. 5 is the gel electrophoresis spectrum after the primer 5 that the embodiment of the present invention 2 provides expands；
Fig. 6 is the SSR finger-print for 6 melon varieties that the embodiment of the present invention 3 provides；
Fig. 7 A-7F is the finger-print QR coding for 6 melon varieties that the embodiment of the present invention 3 provides；
Fig. 8 is the two-dimensional code scanning figure in close No. 1 of the western state that the embodiment of the present invention 3 provides.
Specific embodiment
For further illustrate the present invention to reach the technical means and efficacy that predetermined goal of the invention is taken, below with compared with
Good embodiment, to specific embodiment, technical solution, feature and its effect applied according to the present invention, detailed description is as follows.Under
Stating the special characteristic, structure or feature in multiple embodiments in bright can be combined by any suitable form.
Embodiment 1 (extracts muskmelon DNA using CTAB method)
A. 0.2~0.5g spire is taken, is set in mortar, sufficient liquid nitrogen grinding is added, until at powdered；
B. powder is transferred in 2ml centrifuge tube rapidly；760 μ lCTAB Extraction buffer 100mmol/ are added in Guan Zhongyi
LTris (pH8.0), 1.4mol/LNaCl, 50mmol/L EDTA, 2%CTAB, 2%PVP, 20 μ l beta -mercaptoethanols are sufficiently mixed
It is even；
Pipe lid is opened and closed and shakes 2~3 times during water-bath by c.65 DEG C water-bath 30min；
D. it is cooled to room temperature, adds 24: 1 isometric (V/V) chloroforms：Isoamyl alcohol, about 900 μ l are mixed；
E. room temperature is centrifuged, 12000rpm, 15min；
F. 2ml centrifuge tube is taken, supernatant (about 750 μ l) is transferred to wherein；
G. it is added and the isometric chloroform of supernatant：Isoamyl alcohol (V/V 24:1) it, reverses centrifuge tube for several times, mixes；
H. room temperature is centrifuged, 12000rpm, 15min；
I. step 1~2 time f~h are repeated, until organic phase and the disappearance of water phase intersection white impurity；
J. 2ml or 5ml centrifuge tube is taken, supernatant (about 650 μ l) is transferred to wherein；2 precooled × supernatant fluid is added
Long-pending dehydrated alcohol or isometric isopropanol, gently Spin tubes, are then allowed to stand precipitating DNA, have cotton-shaped DNA to occur in pipe；
K. 1.5ml centrifuge tube is taken, 70% ethyl alcohol of 1ml is added, DNA is chosen into washing wherein, is shaked gently, outwells 70%
Ethyl alcohol, 1ml dehydrated alcohol, which is added, washed once, and outwells dehydrated alcohol, blots dehydrated alcohol with pipette tips.It is placed on superclean bench
On, blower opening 10~15 minutes are clean to ethyl alcohol volatilization；
L. 500 μ l TE [10mmol/LTrisHCl (pH8.0), 1mmol/L EDTA (pH8.0)] dissolving DNA is added；
M. it after DNA is completely dissolved, is added 1 μ l10mg/mL RNA enzyme (final concentration is about 20 μ g/mL).37 DEG C incubate 30
~60min；
N. isometric chloroform can be added again：Isoamyl alcohol (V/V 24:1), reverse centrifuge tube for several times, mix, room temperature from
The heart, 12000rpm, 10min take 1.5ml centrifuge tube, supernatant are transferred to wherein；
O. the dehydrated alcohol of 2 × volume of addition pre-cooling, 1/10 volume 3mol/L NaAc (pH5.2), -20 DEG C of placements 1~
2hr is stayed overnight, and precipitates DNA；
P. 0.5ml centrifuge tube is taken, with 70% ethyl alcohol of 0.5ml and dehydrated alcohol, washing DNA is each primary, blots nothing with pipette tips
Ethyl alcohol.It is sufficiently dry on superclean bench；
Q. 50~100 μ l TE [10mmol/L TrisHCl (pH8.0), 1mmol/L EDTA (pH8.0)] dissolution is added
DNA；
R. DNA sample is dispensed, for a part for using in the recent period, a part of long-term preservation is placed in -20 DEG C of preservations.
By 10 single plants of difference of same muskmelon, (Ru Xizhou is No. 1 close, western state is No. 3 close, western state is No. 25 close, western state close 17
Number, 10 cultivation single plants in western state close No. 21 or western state close 29) young leaflet tablet mixing, gene pool is established, using above-mentioned CTAB method
Extract respective genomic DNA；The integrality and concentration of DNA fragmentation are examined using agarose electrophoresis；Utilize ultraviolet spectrometry light
OD is surveyed on degree meter230、OD260、OD280, calculate OD260/OD230And OD260/OD280, detect DNA purity；If OD260/OD280Ratio exists
Between 1.60-1.90, OD260/OD230It can be used for PCR amplification greater than 2.0, otherwise need to be further purified.
Embodiment 2 (PCR amplification)
(Ru Xizhou is No. 1 close, western state is No. 3 close, western state is No. 25 close, western state is No. 17 close, west by the muskmelon DNA extracted with embodiment 1
State it is close No. 21 or western state it is close 29) for amplification template, be utilized respectively primer 73 (C30), primer 47 (MU5499), primer 14
(MU5554-1), primer 53 (SSR04219) or primer 5 (SSR12083) expand the DNA of above-mentioned 6 melon varieties；Draw
Object is as shown in table 1 to information, and PCR is shown in reaction system such as table 2；
1. muskmelon SSR primer system of table
Table 2.PCR reaction system
| Reaction system | Volume (20 μ L) |
| ddH2O | 7.9μL |
| DNA profiling | 1.5μL |
| Primer P1, P2 | Each 0.3 μ L |
| 10×Buffer | 10.0μL |
PCR response procedures：94 DEG C of initial denaturation 5min；94 DEG C of denaturation 15s, appropriate annealing temperature 15s, 72 DEG C of extension 30s are followed
Ring 35 times；72 DEG C of extension 4min；
After to PCR amplification, using liquid-transfering gun, clicks and enters on 8% non-deformed polyacrylamide gel and carry out vertical electricity
Swimming, electrophoresis finishes removes gel progress silver staining colour developing immediately, and running gel result is photographed to record with digital camera, such as Fig. 1-Fig. 5.
Embodiment 3 (building muskmelon SSR finger-print)
The genomic DNA (total DNA) of 18 muskmelon materials shown in the 5 pairs of SSR primer pair tables 3 designed using embodiment 2
It is expanded, the amplification of each primer can obtain different banding patterns, and (different numbers represents different banding patterns, in Fig. 6
Primer 73 in the banding pattern that successively occurs use 1,2,3,4,5 indicate that is, 1,2,3,4,5 respectively represent 5 different bands respectively
Type), therefore utilize the finger-print of available 18 materials of primer pair 73,47,14,53,5, the wherein western state of new varieties close 1
Number, western state is No. 3 close, western state is No. 25 close, western state is No. 17 close, the finger-print in western state close No. 21 and western state close 29 is as shown in fig. 6, figure
1-18 in 6 respectively represents that close No. 1 female parent in western state, western state be No. 1 close, close No. 1 male parent in western state；Close No. 3 female parents in western state, western state close 3
Number, close No. 3 female parents in western state；Close No. 25 female parents in western state, western state be No. 25 close, close No. 25 male parents in western state；Western state close No. 17 female parents, Xi Zhou
Close No. 17, close No. 17 male parents in western state；Close No. 21 female parents in western state, western state be No. 21 close, close No. 21 male parents in western state；Western close No. 29 mothers in state
This, western state is No. 29 close, close No. 29 male parents in western state.
3. muskmelon material of table
It can be seen that by Fig. 6, western close No. 1 finger-print code in state is 33131, close No. 3 finger-print codes in western state are 33253, Xi Zhou
Close No. 25 finger-print codes are 33311, close No. 17 finger-print codes in western state are 53215, close No. 21 finger-print codes in western state are 15511 and west
Close No. 29 finger-print codes in state are 36611.
Finger-print QR coding application two dimensional code editing machine is encoded, by the material name of each new varieties (above-mentioned 6 kinds)
Title, type, Plant Taxonomy and SSR finger-print code typing software together form finger-print QR coding, such as Fig. 7 A-
Shown in 7F, two dimensional code barcode scanning figure is as shown in Figure 8.
Above-mentioned 5 pairs of primers of the invention are western state is No. 1 close, western state is No. 3 close, western state is No. 25 close, western state is No. 17 close, Xi Zhoumi
Identification result between No. 21 and western state close 29 becomes apparent from.
Place, those skilled in the art can not select from the prior art to the greatest extent in the embodiment of the present invention.
Disclosed above is only a specific embodiment of the invention, but scope of protection of the present invention is not limited thereto, is appointed
What those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, answer
It is included within the scope of the present invention.Therefore, protection scope of the present invention should be with above-mentioned scope of protection of the claims
It is quasi-.
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Claims (3)
1. a kind of SSR molecular marker system of muskmelon, which is characterized in that the molecular labeling system includes 5 primer pairs；
The entitled C30 of primer pair 1, nucleotides sequence are classified as SEQ ID NO.1 and SEQ ID NO.2；
The entitled MU5499 of primer pair 2, nucleotides sequence are classified as SEQ ID NO.3 and SEQ ID NO.4；
The entitled MU5554-1 of primer pair 3, nucleotides sequence are classified as SEQ ID NO.5 and SEQ ID NO.6；
The entitled SSR12083 of primer pair 4, nucleotides sequence are classified as SEQ ID NO.7 and SEQ ID NO.8；
The entitled SSR04219 of primer pair 5, nucleotides sequence are classified as SEQ ID NO.9 and SEQ ID NO.10.
2. a kind of SSR molecular marker system of muskmelon as described in claim 1, which is characterized in that the new varieties of the muskmelon
For western state is No. 1 close, western state is No. 3 close, western state is No. 25 close, western state is No. 17 close, western state close No. 21 or western state close 29.
3. a kind of application of the SSR molecular marker system of muskmelon of any of claims 1 or 2 in identification melon variety.
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| CN201810763805.XA CN108841989B (en) | 2018-07-12 | 2018-07-12 | SSR molecular marker system of muskmelon and application thereof |
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| CN108841989B CN108841989B (en) | 2021-09-21 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112961931A (en) * | 2021-03-02 | 2021-06-15 | 宁波市农业科学研究院 | Rapid identification method for purity of Yongtian No.5 melon seeds |
-
2018
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Non-Patent Citations (2)
| Title |
|---|
| 熊姜玲: "甜瓜株型性状的遗传分析与分子标记", 《中国优秀硕士学位论文全文数据库》 * |
| 王美荣: "甜瓜育种亲本材料遗传多样性及群体结构的SSR标记分析", 《华北农学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112961931A (en) * | 2021-03-02 | 2021-06-15 | 宁波市农业科学研究院 | Rapid identification method for purity of Yongtian No.5 melon seeds |
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